Prosiding Persidangan Industri Herba 2015

Transcription

FRIM Proceedings No. 7
PROSIDING
PERSIDANGAN
INDUSTRI HERBA
PUTRAJAYA
Editor
O Asiah
HF Lim
BJ Chee
M Nik Musaadah
M Mastura
K Getha
MGH Khoo
P Mazura
S Vimala
FRIM Proceedings No. 7
Prosiding Persidangan Industri Herba 2015
MEMPERKASA JALINAN INDUSTRI HERBA KE ARAH
TRANSFORMASI EKONOMI
3─5 November 2015
Putrajaya
Editor
O Asiah
HF Lim
BJ Chee
M Nik Musaadah
M Mastura
K Getha
MGH Khoo
P Mazura
S Vimala
2015
© Institut Penyelidikan Perhutanan Malaysia 2015
Segala pertanyaan hendaklah dikemukakan kepada:
Ketua Pengarah
Institut Penyelidikan Perhutanan Malaysia
52109 Kepong
Selangor Darul Ehsan
Malaysia
Tel: 603-62797000
Faks: 603-62731314
http://www.frim.gov.my
ISBN 978-967-0622-47-7
MS ISO 9001:2008
Diset dalam Calibri 11/12
Dihasilkan di Malaysia oleh Institut Penyelidikan Perhutanan Malaysia, Kepong
KANDUNGAN
Prakata
ix
Pembentangan Pleno
1
Memartabatkan Pengetahuan Tradisi: Antara Khazanah Warisan
Bangsa dan Sumber Kekayaan Baharu Negara
H Norini
2
Tongkat Ali: From Lab to Market
TA Tengku Shahrir & R Haliza R.
4
Pembentangan Lisan
5
Hala Tuju Industri Herba di Bawah NKEA
A Roziah & R Farhan
6
Peranan FRIM Dalam Memperkasa Industri Herba di Malaysia
MA Rasadah
13
Menyusun Strategi Berpandukan Fakta Rantaian Nilai Industri Herba
M Marzalina, AB Ariff Fahmi, AR Rohana & R Rosniza
21
Pendokumenan Pengetahuan Tradisi Melayu Berkaitan Tumbuhan
Ubatan di Semenanjung Malaysia
M Nik Musaadah, M Dionysia, MS Abdul Hayat, Z Nurul Husna, J
Fadzureena, B Intan Nurulhani et al.
26
Peralatan Dalam Perubatan Tradisional Melayu: Khazanah Ilmu dan
Peradaban Warisan Bangsa
A Ida Farah, AZ Wan Nurul Syafinaz, Z Nurasyikin & SM Ami Fazlin
31
Penyelidikan dan Pembangunan serta Pengkomersialan (R&D&C)
Produk Berasaskan Tanaman Ubatan dan Beraroma di MARDI
WM Wan Zaki, Z Mohd Hafizudin, MA Mohd Shukri, A Norzihan, J Indu
Bala, S Roowi & H Norma
36
Kajian Rantaian Nilai Industri Herba Terpilih di Semenanjung
Malaysia
AR Rohana, AB Ariff Fahmi, Z Nur Fazreen, A Nur Syazni, S Siti
Zubaidah, R Rosniza et al.
44
iii
Mengangkat Tongkat
Pengkomersialan
MY Awang Ahmad
Ali
ke
Mata
Dunia:
Pengalaman
51
Analysing Malay Medical Manuscript MSS2999: the Findings and the
Hope for the Future
H Abdul Ghani
56
Commercialized Innovative Herbal Products From UPM
M Suhaila & MN Fatihanim
59
Development of Herbal Product for Cancer Therapy
AH Loiy Elsir, H Suzana, AM Aman Shah & AM Amin Malik Shah
65
Pembentangan Poster
71
Saringan Fitokimia dan Aktiviti Anti-Radang Tumbuhan Ubatan
Terpilih Warisan Orang Asli Semelai, Pos Iskandar, Bera, Pahang
MA Adiana, H Nuziah, J Fadzureena, M Nik Musaadah, Y Norshakila, AL
Tan et al.
72
Kandungan Fitokimia Mas Cotek Berlainan Aksesi
J Samsiah, D Thiyagu, MS Rosalizan, M Osman, S Noor Safuraa & B Siti
Salwah
78
Penentuan Julat Eurycomanone Dalam Produk Tongkat Ali Melalui
Kaedah Kromatografi Cecair Ultraprestasi (UPLC)
A Mohd Radzi, SK Ling, L Abd Rashid & MT Nurzalikha
84
Penilaian Kandungan Fitokimia dan Aktiviti Sitotoksiti Spesies
Terpilih Tumbuhan Ubatan Berasaskan Pengetahuan Tradisional
Subetnik Jahai/Temiar di Kampung Air Banun, Gerik, Perak
MA Adiana, S Rohana, H Nuziah, J Fadzureena, M Nik Musaadah, AL
Tan et al.
90
Asas Molekul Aktiviti Anti-Malaria Kurkumin
A Amatul Hamizah, S Suhaini, E Noor & MS Hasidah
96
Mekanisme Tindakan Sebatian Bioaktif Anti-Malaria Dalam Paku
Pakis Ubatan, Gleichenia truncate
S Suhaini, E Noor & MS Hasidah
101
iv
Pemilihan dan Penghasilan Baka Limau Purut (Citrus hystrix)
Bermutu Tinggi
MA Farah Fazwa, J Mailina, MA Nor Azah, M Nur Nazihah, A Mohd.
Zaki, SB Syafiqah Nabilah et al.
106
Pengeluaran dan Penilaian Minyak Daun Kayu Manis Malaysia
H Norma, N Noor Ismawaty, M Erny Sabrina, AR Zuraida, EA Engku
Hasmah, M Razali et al.
113
Pembiakan Aksesi Terpilih Kacip Fatimah (Labisia pumila var. alata)
Melalui Kaedah Keratan dan Penilaian Pengeluaran Pucuk Pada
Peringkat Tapak Semaian
M Nur Nazihah, MA Farah Fazwa, S Norhayati, SB Syafiqah Nabilah, L
Mohd Asri & Z Mohd Zaini
118
Serangan Atteva sciodoxa (Ulat Harimau) Pada Tanaman Tongkat Ali
di Hutan dan Ladang Semenanjung Malaysia
WA Wan Muhd Azrul, A Mohd Farid, AS Sajap, S Tosiah & M Patahayah
125
Produk Herba dan Trend Pencemaran Mikroorganisma
Y Norulaiman, BK Ong, MA Nor Azah, MH Nurhazwani, A Nor Hayati &
MS Amira Rina Nurdiana
132
Rumpai Miang Mexico, Ancaman Melebihi Manfaat
AK Azimah, MA Anis Fadzilah, M Dilipkumar & AH Muhammad Saiful
138
Konservasi Secara Ex Situ bagi Kacip Fatimah Varieti Lanceolata
SB Syafiqah Nabilah, MA Farah Fazwa, S Norhayati & M Nur Nazihah
146
Tapak Warisan FRIM: Potensi Sebagai Tapak Konservasi Ex Situ
Terbesar Bagi Germplasma Tumbuhan Ubatan di Semenanjung
Malaysia
A Noorsiha, AM Ainnur Amira, M Kamariah, MK Mohd Haire, B Nuranis
Suraya & F Nur Fairuz
153
Penggunaan Tumbuhan Dalam Kehidupan Orang Asli Kumpulan
Senoi di Semenanjung Malaysia: Khususnya Subetnik Temiar dan
Semai
AL Tan, BJ Chee, MN Madihah, M Mastura, B Intan Nurulhani, M
Badariah et al.
159
Penggunaan Herba Dalam Perbidanan Tradisional Melayu Selepas
Bersalin di Zon Tengah, Semenanjung Malaysia
A Siti Khairul Bariyyah
164
v
Trend Penggunaan 18 Spesies Tumbuhan Ubatan di Bawah Program
NKEA di Kalangan Pengamal Perubatan Tradisional Melayu di
Semenanjung Malaysia
M Dionysia, MS Abdul Hayat, M Nik Musaadah, B Intan Nurulhani, MN
Madihah, Z Nurul Husna et al.
170
Amalan Perubatan Tradisional Melayu di Semenanjung Malaysia:
Adakah Masih Relevan Pada Masa Kini ?
MS Abdul Hayat, M Dionysia, M Nik Musaadah, B Intan Nurulhani, Z
Nurul Husna, J Fadzureena et al.
175
Peralatan Penyediaan Ubatan Dalam Kaedah Rawatan Perubatan
Tradisional Melayu
AZ Wan Nurul Syafinaz, A Ida Farah, SM Ami Fazlin & BA Siti Khairul
180
Physico─Chemical Properties of Spray Dried Clinacanthus nutans
Extract
A Mohd. Suhaimi & AL Abd. Rahman
185
Extraction of Antioxidants from Leaves of Clinacanthus nutans
(Burm.F.) Lindau: Effects of Extraction Method and Solvent
NA Karim & II Muhamad
192
Anti-Proliferative Effect of Clinacanthus nutans on Ovarian, Breast
and Colorectal Cancer Cell Lines
Y Nurhanan Murni, SK Ling, MM Siti Syarifah, A Zunoliza & MY Nor
Afiedatul
200
Detection of Irradiated Herbs Using Photo-Stimulated Luminescence
Technique (PSL)
AR Ros Anita, O Zainon, A Foziah & H Zainab
209
Optimization of Antioxidant Activity in Clinacanthus nutans (Belalai
Gajah/Sabah Snake Grass)
HS Kong, KH Musa & NA Sani
214
Effects of Equal Doses of Sulforaphane, Curcumin and Quercetin on
Heme Oxygenase 1 Gene Expression in Mice Liver
A Abdullah, N Alrawaiq & A Elbadri
218
Evaluation of Antimicrobial and Antioxidant Property of Anethum
graveolens Leaf Extracts
MN Mohd Effendi & AS Norrakiah
223
vi
Asam Gelugur Powder Rich in HCA (Hydroxycitric Acid): a Potential
Crop for Weight Management
HZ Umi Kalsum, HA Hashimah, A Sharizan, A Mohamed Nazim, M
Aida, S Nor Fadhilah et al.
228
Toxicology Study of Hibiscus sabdariffa L. Leaves Extract on Normal
Sprague-Dawley Rats
H Hasnisa, M Syahida, H Hadijah, Z Kharis, A Sharizan, D Mohd Nazrul
Hisham et al.
233
Clinnacanthus nutans L.: Safety and Toxicity Study
M Syahida, H Hasnisa, AG Rosnani, Z Kharis, DMN Hisham, R Suri et al.
240
In Vitro Toxicological Evaluation of 50 Methanol Extracts From
Traditional Medicinal Plants Used by the Orang Asli
K Mary, S Rohana, M Nik Musa’adah, J Fadzureena, M Adiana, H
Nuziah et al.
246
Toxicity Study of Sarawak Wild Pepper Root (Piper arborescens)
HP Chua, M Syahida & D Nicholas
251
In Silico Prediction of Drug Likeness and ADMET Properties of Some
Centella Compounds
O Asiah, J Mohd Haffiz, MM Siti Syarifah, MA Nordatiakma, Y
Nurhanan, H Lili Sahira et al.
257
Nutrigenomic Effects of Curculigo latifolia on Type 2 Diabetic Model
I Nur Akmal & I Maznah
264
Protein Expression Pattern in Swietenia macrophylla Seed
A Noraliza, MA Nor Datiakma, M Norwati, WK Wan Mohd Aizat, MN
Normah, H Nor Asmah et al.
274
Proteome Profiles of Seeds from Swietenia macrophylla After Cold
Storage
A Noraliza, MA Nor Datiakma, Z Fadzlinah, M Norwati, WK Wan Mohd.
Aiza & MN Normah
279
Effect of pH on Adsorption of Organic Acid and Phenolic Compounds
in Noni (Morinda citrifolia L.)
H Haslaniza, WA Wan Yaakub & MY Maskat
283
vii
Effect of Organic-Based Fertilizer Rate and Planting Distance on
Biomass Yield of Belalai Gajah (Clinacanthus nutans)
AG Rosnani & M Syahida
289
The Effect of Packaging Technique on Quality of Dried Mas Cotek
(Ficus deltoidea)
MS Rosalizan, D Thiyagu & D Mohd Nazrul Hisham
296
Potential of Medicinal Plants Used by the Jakun People as
Antituberculosis Agents
S Siti Fatimah, M Maryati & AB Mohd Fadzelly
303
Medicinal Plants Used for Women’s Healthcare Among the Jakun
Community in Kg. Peta: a Preliminary Study
I Nur Amalina, M Maryati & AB Mohd. Fadzelly
309
Kaempferia L. : Herbs and Ornamental Potential of Zingiberaceae
Species
R Izlamira, MA Zulkhairi, MZ Nurin Izzati & B Suryanti
313
Distribution of Herbs Cultivation in Peninsular Malaysia
A Nur Syazni, AR Rohana, AB Ariff Fahmi, Z Nur Fazreen, S Siti
Zubaidah & M Marzalina
318
Halal Herbal Products Integrity Risk Through Supply Chain: a
Conceptual Study
R Salini Devi & K Nitty Hirawaty
324
Consumer Preferences in Selecting Herbal Products
S Siti Zubaidah, AR Rohana, AB Ariff Fahmi, & A Nur Syazni
331
Consumers’ Perception Towards Local Herbal Supplement Products
NH Kamarulzaman & SM Talib
336
The Importance of Long-Term Relationship for the Sustainability of
Malaysian Herbal Industry
H Nurul Syahira, MN Nolila, M Norsida & M Zainal Abidin
343
viii
PRAKATA
Industri herba yang melibatkan penggunaan bahan mentah berasaskan
tumbuhan ubatan dan beraroma dikenalpasti mempunyai masa hadapan yang
cerah dan boleh dibangunkan sebagai satu sektor ekonomi untuk sumber
kekayaan baharu negara. Pembangunan industri herba pula telah menjadi satu
agenda penting yang telah diketengahkan dalam sektor pertanian Bidang
Ekonomi Utama Negara (NKEA). Malah, pembangunan industri herba telah
dijadikan satu cabang utama penggerak kepada Dasar Agromakanan Negara.
Industri herba Malaysia sedang pesat membangun, namun jika dibandingkan
dengan penghasilan produk herba negara-negara lain di pasaran dunia, ianya
masih tertumpu kepada produk akhir bernilai rendah atau pemprosesan hanya
ke peringkat produk pertengahan. Dalam usaha untuk menembusi pasaran
global dan meningkatkan sumbangan industri berasaskan herba kepada
ekonomi negara, produk sedia ada dan baru perlu ditingkatkan dari segi kualiti,
nilai dan keselamatan. Dalam masa yang sama, industri herba turut berdepan
dengan cabaran untuk meningkatkan pendapatan kumpulan sasar supaya
jurang pendapatan antara sektor ekonomi dan antara kawasan bandar dan luar
bandar dapat dirapatkan.
Oleh yang demikian, penganjuran Persidangan Industri Herba 2015 yang
bertema “Memperkasa Jalinan Industri Herba Ke Arah Transformasi Ekonomi”
amat bertepatan. Kini tiba masanya hubungan erat dan kukuh dibina dan
dijalin pada semua peringkat melalui interaksi para penyelidik, ahli akademik,
wakil-wakil pengiat industri herba, pengusaha tanaman herba serta
penglibatan komuniti pemilik pengetahuan tradisi untuk mengerakkan industri
ini ke tahap yang lebih tinggi.
Prosiding yang mengandungi kompilasi kertas kerja mengenai dasar, akta dan
strategipengetahuan tradisi, penyelidikan dan pembangunan serta pasaran
herba dapat memberi maklumat berguna dan menjadi rujukan penting bagi
pihak yang berkepentingan dalam penyelidikan mahupun industri herba. Saya
berharap usaha pengumpulan maklumat mengenai herba dan hasilan semula
jadi daripada segala aspek akan diteruskan agar hasrat dan aspirasi Kerajaan
Malaysia yang mengenal pasti industri herba sebagai sumber pertumbuhan
ekonomi Negara di bawah NKEA dalam program transformasi ekonomi (ETP)
akan terlaksana.
DATO’ DR ABD LATIF MOHMOD
Ketua Pengarah
Institut Penyelidikan Perhutanan Malaysia
ix
PEMBENTANGAN
PLENO
1
MEMARTABATKAN PENGETAHUAN TRADISI: ANTARA KHAZANAH
WARISAN BANGSA DAN SUMBER KEKAYAAN BAHARU NEGARA
H Norini
Institut Penyelidikan Perhutanan Malaysia (FRIM), 52109 Kepong, Selangor
Tel: 03-6279 7540
E-mel: [email protected]
ABSTRAK
Sebagai sebuah institut penyelidikan perhutanan tersohor di dunia, FRIM tidak
ketinggalan dalam memartabatkan pengetahuan tradisi (traditional knowledge)
pendeknya TK yang ada hubung kaitnya dengan biodiversiti dan hutan bagi
beberapa etnik terpilih di Semenanjung Malaysia. TK sendiri mempunyai
pengertian amat luas, meliputi pengetahuan yang disampaikan melalui
penceritaan, amalan harian, undang-undang adat, pantang larang, budaya,
upacara dan kepercayaan yang diwarisi turun temurun. Apabila TK itu
diturunkan dari satu generasi ke satu generasi secara automatik ianya menjadi
khazanah warisan. Sebenarnya, usaha memartabatkan TK ini bukan satu isu
baharu. Apa yang berbeza ialah cara pendekatan yang digunakan bagi
memartabatkan TK ini agar dapat mengangkatnya ke tahap yang lebih tinggi,
disegani dan disanjung tinggi serta menjadi kebanggaan bukan hanya oleh
etnik berkaitan malahan masyarakat Malaysia yang majmuk. Menyingkap
kembali sejarah, penyelidikan terhadap herba/tumbuhan ubatan telah
dilaksanakan oleh FRIM semenjak 1990an lagi. Hasil penyelidikan bukan hanya
menyumbang kepada peningkatan ilmu pengetahuan dalam bidang berkenaan
malahan juga jika didapati berpotensi turut diusahakan untuk dikomersialkan.
Rata-rata juga maklum bahawa herba/tumbuhan ubatan mempunyai potensi
untuk ditransformasikan sebagai nadi penggerak pertumbuhan bioeknomi
baharu. Tidak juga dinafikan keupayaan mentransformasikan pengetahuan
herba/tumbuhan ubatan ini telah menjadikan jika tidak semua pengusaha
famaseutikal, nutraseutikal dan kosmeseutikal kaya raya tetapi tidak empunya
ilmu. Menyedari hakikat bahawa Malaysia sebagai sebuah negara yang telah
menandatangani dan terikat dengan Konvensyen Kepelbagaian Biologi (KKB)
pada 1994, FRIM telah mengorak langkah pada 2007 untuk mendokumenkan
TK Orang Asli berkaitan tumbuhan ubatan di Semenanjung Malaysia. Projek
penyelidikan ini bukan setakat mendokumenkan sahaja malahan termasuk juga
memberikan latihan kepada setiap subetnik dan menganalisis tumbuhan
ubatan terpilih untuk menentu sahkan TK yang didokumenkan. Usaha kami
diperteguhkan lagi apabila kami terus mara ke hadapan dan berjaya
menghasilkan produk prototaip pertama bagi subetnik Semelai dengan nama
Semelai’s Secret. Apa lagi yang menarik mengenai usaha penyelidikan kami
ialah dalam semua pelaksanaan aktiviti projek, pendekatan prior informed
2
consent digunakan bagi memastikan sokongan padu setiap subetnik. Ternyata
pendekatan ini membuahkan hasil yang amat lumayan. Usaha
mendokumenkan TK berupaya mengekalkan khazanah warisan sementara
menghasilkan prototaip dan kemudian mengkomersialkan pula boleh
menjadikan herba/tumbuhan ubatan sebagai sumber kekayaan baharu negara.
Paling penting, hasil mahsul mengkomersialkan produk dikongsi kembali
dengan empunya ilmu. Dengan tekad membara kami terus melangkah, kali ini
dengan mendokumentasikan TK Orang Melayu pula pada pertengahan 2013.
Kami juga sedang berusaha membangunkan kerangka bagi perkongsian faedah.
Sebenarnya, terlalu banyak yang ingin dilaksanakan. Dengan dana yang terhad
kami cuba melangkah setapak demi setapak, tetapi kami yakin langkah kami
mampu menggegarkan dunia kerana hingga kini tidak ada mana-mana institusi
yang mengutarakan pakej penyelidikan yang sedemikian.
Kata kunci: traditional knowledge, herba/tumbuhan ubatan, khazanah warisan,
prior informed consent, sumber kekayaan, perkongsian faedah, Orang Asli
3
TONGKAT ALI: FROM LAB TO MARKET
TA Tengku Shahrir & R Haliza
BIOTROPICS MALAYSIA BERHAD, Lot 21, Jalan U1/19, Section U1, HicomGlenmarie Industrial Park, 40150 Shah Alam, Selangor
Tel: 03-5565 5600 Fax: 03-5565 5602 E-mail:[email protected]
ABSTRACT
Amongst Malaysia’s rich heritage of biodiversity and traditions of medicinal
knowledge, Tongkat Ali for men’s health is arguably, currently, the most sought
after in the market, locally and internationally. It is now widely known
internationally as Malaysian Ginseng, in-spite of its existence and use in
Indonesia, Thailand, Philippines, and Vietnam. For many reasons, Tongkat Ali is
precious to the local industry having the most established chain of harvesters
and manufacturers, as well as being the most scientifically advanced product
through the efforts of our scientists.
The paper shares the experience of Biotropics Malaysia Berhad, as one of the
many vehicles in Malaysia, in the country’s collective effort to establish the
presence of Tongkat Ali, as a new health product in the international market.
The paper discusses the research works that have been undertaken not only to
meet the different requirements and restrictions by the health regulators in
the various countries, but also to meet the more sophisticated demands of the
modern and educated consumers of health products worldwide. The paper
looks at the commercialisation initiatives undertaken to introduce this
wondrous Malaysian herb in global platforms, and educating the consumers
on its numerous health benefits. The paper also shares some challenges faced
by Biotropics in this long, arduous but fulfilling journey, and tries to extract
some learning from the experiences that may be useful to other players who
may wish to tread the same journey.
Keywords: tongkat ali, Malaysian ginseng, health product, men’s helath
4
PEMBENTANGAN
LISAN
5
HALA TUJU INDUSTRI HERBA DI BAWAH NKEA
A Roziah & R Farhan
Bahagian Pembangunan Herba (HDD), Galeria PJH, Aras 2, No. 29. Jalan P4W,
Persiaran Perdana, Presint 4, 62100, Putrajaya
Tel: 03-8894 3102 Faks: 03-8894 3125
ABSTRAK
Program Bidang Ekonomi Utama Negara (NKEA) merupakan pemacu kegiatan
ekonomi yang berpotensi memberi sumbangan penting kepada pertumbuhan
ekonomi yang diterajui oleh sektor swasta dan disokong oleh pihak Kerajaan
yang bertindak sebagai pemudahcara dan fasilitator. Kerajaan telah
mengumumkan Program Transformasi Ekonomi (ETP) pada Oktober 2011 yang
memfokuskan kepada bidang–bidang utama yang memacu ekonomi negara
dan menjadikan Malaysia ke arah negara yang berdaya saing. Sebanyak 12
NKEA menjadi teras utama Program Transformasi Ekonomi (ETP) dan NKEA
Pertanian adalah merupakan salah satu teras penting yang diterajui oleh
Kementerian Pertanian dan Industri Asas Tani (MOA). Sektor Pertanian
memainkan peranan penting dalam pembangunan ekonomi Malaysia di mana
ianya dapat mewujudkan peluang pekerjaan dan meningkatkan pendapatan
penduduk luar bandar serta menjamin keselamatan makanan negara.
Matlamat utama NKEA Pertanian adalah untuk mentransformasikan sebuah
sektor yang secara tradisinya berskala kecil dan berasaskan pengeluaran
kepada sebuah industri perniagaan asas tani berskala industri. Ianya
dilaksanakan bagi menyumbang kepada pertumbuhan ekonomi yang lebih
mapan dan berdaya saing. NKEA pertanian ini adalah merupakan model
pemusatan pasaran bersepadu yang tertumpu kepada ekonomi pertanian dan
penyepaduan rantaian nilai. Strategi transformasi ini mengandungi tumpuan
kepada kelebihan bersaingan, menembusi pasaran premium, memastikan
dasar keterjaminan makanan yang selaras dengan peningkaan nilai
pertambahan pendapatan Negara (PNK) dan memperkembangkan penglibatan
dalam rantaian nilai sektor pertanian serantau.
Berdasarkan teras-teras di dalam NKEA tersebut, Projek Keutamaan Permulaan
(EPP) telah diperkenalkan melalui Makmal NKEA pada tahun 2010 yang
membangunkan 16 EPP utama bagi mamacu ekonomi negara. Inisiatif EPP ini
bertujuan bagi mencapai PNK berjumlah RM28.9 bilion menjelang 2020. Selain
daripada itu, 109,335 tambahan peluang pekerjaan juga diharap dapat dicapai
sepanjang tempoh tersebut. EPP1 iaitu membuka nilai daripada biodiversiti
Malaysia menerusi pengeluaran produk herba bernilai tinggi yang merupakan
salah satu EPP yang dihasilkan menerusi NKEA tersebut dan Bahagian
6
Pembangunan Herba (HDD) merupakan badan penggerak utama dalam
pelaksanaan program EPP1 ini.
Kata kunci: NKEA, Bahagian Pembangunan Herba (HDD), inisiatif, industri
herba
PENGENALAN
Fokus utama industri herba adalah untuk memperkasa kualiti produk bagi
menembusi pasaran eksport global dalam produk nutraseutikal dan perubatan
botaniKal. Di dalam NKEA Pertanian, EPP1 Produk Herbal Bernilai Tinggi
dijangka memberi nilai pasaran yang besar iaitu sebanyak AS$93 bilion pada
tahun 2015 dan dijangka meningkat kepada AS$107 bilion pada tahun 2017.
Faktor utama pertumbuhan positif nilai pasaran herba ini adalah berdasarkan
manfaat dan khasiat tersendiri herba yang diamalkan sejak dahulu lagi yang
secara tidak langsung mencetuskan kesedaran ke atas penggunaan herba di
kalangan rakyat Malaysia.
Jemaah Menteri telah meluluskan penubuhan Majlis Pembangunan
Herba Negara pada 9 Februari 2011 yang secara langsung merealisasikan
penubuhan Bahagian Pembangunan Herba (HDD) sebagai agensi pelaksana.
Bahagian ini telah diberi mandat untuk mengawalselia, melaksana dan
memantau projek herba bagi EPP1 yang asalnya diberi tanggungjawab utama
untuk membangunkan industri herba bagi lima herba utama negara iaitu
tongkat ali, kacip fatimah, misai kucing, dukung anak dan hempedu bumi ke
arah menghasilkan produk herba bernilai tinggi. Jumlah tanaman yang
tersenarai di dalam EPP#1 pun telah diperluakan kepada 18 jenis (Rajah 1) dan
berkemungkinan besar senarai ini akan ditambah lagi pada masa akan datang.
Kini, HDD memainkan peranan penting sebagai badan pengkoordinasi
(coordinating
body)
antara
kerajaan,
industri
serta
institusi
penyelidikan/institusi pengajian tinggi bagi tujuan untuk meningkatkan kualiti
dengan menerapkan sains ke dalam produk herba negara. HDD seterusnya
akan menyelaras peningkatkan kualiti produk herba negara bersama usahawan
tempatan yang diharap akan mampu dipertingkatkan lagi nilainya ke tahap
yang lebih tinggi.
7
Rajah 1. 18 Fokus Herba di Bawah NKEA EPP#1 Produk Herba Bernilai Tinggi
SKOP PELAKSANAAN PROJEK PEMBANGUNAN HERBA DI BAWAH HDD
Skop Inisiatif EPP1 dirangka bagi memenuhi rantaian keperluan penghasilan
produk dalam industri herba bermula di peringkat huluan hingga hiliran.
Inisiatif-inisiatif ini akan mencungkil kepelbagaian bio negara yang luas dan
unik sebagai sebuah pusat produk herba yang berpotensi. Ia bertujuan untuk
memperkukuhkan kualiti dan usaha memasarkan produk nutraseutiKal dan
ubat berasaskan botani bagi menembusi pasaran eksport global. Inisiatifinisiatif yang dirangka oleh HDD dalam memacu ekonomi pasaran herba adalah
seperti berikut:
Klaster Herba
Pembangunan klaster herba di peringkat huluan yang dibangunkan oleh
Jabatan Pertanian, Malaysia (DOA) bertujuan untuk memastikan bekalan bahan
mentah mencukupi. Pembangunan yang meliputi kawasan seluas 869.76
hektar di Semenanjung Malaysia yang melibatkan 831 usahawan herba telah
dilaksana bagi membangunkan tanaman herba ini. Kadar insentif teknologi
8
maksimum sebanyak RM14,500/ha diberikan kepada peserta yang melibatkan
pembangunan kawasan dan infrastruktur ladang iaitu pembersihan kawasan,
sistem pengairan dan saliran, jalan dan parit, ladang serta pagar. Projek ini
dilihat berpotensi dalam memenuhi keperluan dan permintaan semasa bahan
mentah yang berkualiti dan konsisten kepada pengusaha-pengusaha tempatan
yang terlibat dengan program NKEA Pertanian untuk sub kumpulan projek
herba negara di bawah pengurusan sistem rantaian bekalan yang konsisten.
Taman Penanaman Herba (Herbal Park)
Objektif penubuhan Taman Penanaman Herba atau Herbal Park ini adalah
untuk penanaman bahan mentah secara komersial yang diterajui oleh Majlis
Pembangunan Koridor Ekonomi Pantai Timur (ECERDC) di Pasir Raja, Chegar
Perah dan Durian Mentangau. Ianya dijangka dapat memenuhi keperluan
industri herba memandangkan tanaman herba yang ditanam secara
berkelompok dan sebagai hab bekalan kepada pengusaha-pengusaha
mendapatkan bahan mentah. Sebanyak tiga syarikat peneraju telah dilantik
bagi mengusaha penanaman ini melalui pemilihan penilaian teknikal yang ketat
di peringkat HDD untuk memastikan projek ini dapat dilaksanakan mengikut
jadual. Model Pelaksanaan Taman Penanaman Herba ini adalah diuruskan
sepenuhnya oleh syarikat peneraju yang dilantik dengan dipantau secara mikro
oleh pihak ECERDC. Selain daripada itu, kaedah pelaksananaan penanaman
secara Satellite Farm di Herbal Park ini juga dibuka kepada petani-petani kecil
di kawasan tersebut. Satelliet Farm ini merupakan salah satu platform bantuan
kepada pengusaha-pengusaha kecil menambah pendapatan dan seterusnya
memangkin ekonomi Industri Kecil dan Sederhana (IKS). Mesyuarat
pemantauan berkala dan lawatan tapak sentiasa dilaksanakan bagi memastikan
projek ini dilaksanakan dengan lancar dan mencapai sasaran yang dikehendaki
oleh Kerajaan.
Skim Geran Penyelidikan NKEA (NRGS)
Skim ini diwujudkan bagi menggalakkan perkembangan industri herba melalui
penyelidikan. Ianya merupakan geran untuk aktiviti pengujian herba yang
merangkumi penyelidikan asas dan gunaan yang boleh menghasilkan output
dalam bentuk teori, konsep, teknologi dan proses berinovatif serta pengujian
pra klinikal dan klinikal demi kemajuan industri herba. Lima kumpulan
Penyelidikan dan Pembangunan (R&D) untuk ‘Cluster of Excellence’ ditubuhkan
untuk menyelaras R&D bagi setiap institusi penyelidikan dan institusi pengajian
tingggi (IPT) serta syarikat peneraju terpilih dalam kluster iaitu:
9
I. Penemuan (Discovery)
Penyelidikan melalui kajian kimia dan penilaian terapeutik ke atas
tumbuhan ubatan dan beraroma bagi mengenalpasti sebatian bioaktif
dan mekanisma tindak balas bahan aktif dalam penghasilan produk
yang berkualiti dan selamat.
II. Penanaman dan agronomi (Agronomy)
Fokus ke atas prolehan bahan mentah yang berkualiti tinggi
merangkumi bidang pembaikbakaan, pengurusan tanaman, penyakit,
perosak dan rumpai serta pengendalian lepas tuai.
III. Teknologi pemprosesan (Processing Technology)
Proses operasi pembuatan sediaan atau produk akhir herba
merangkumi aspek pengeskstrakan bahan mentah yang terpiawai dan
berinovasi dari peringkat makmal ke peringkat skala besar.
IV. Pemiawaian & pembangunan produk (Standardization & Product
Development)
Pemiawaian bahan mentah dan pembangunan produk herba terpiawai
bagi penentuan secara tepat paras kandungan bahan penanda serta
metabolic aktif bagi mematikan ekstrak herba mempunyai kualiti yang
konsisten untuk keberkesanan dan keselamatan produk.
V. Pra Klinikal & Klinikal
Proses pengujian yang dilaksanakan ke atas haiwan yang dinamakan
sebagai praklinikal dan pengujian ke atas manusia dinamakan sebagai
klinikal. Ianya bagi membuktikan keberkesanan dan keselamatan
penggunaan sesuatu produk herba yang dihasilkan dengan mematuhi
Amalan Klinikal baik (Good Clinical Practice).
Setiap kajian penyelidikan ini akan melalui beberapa proses penilaian
teknikal yang diketuai oleh ketua klaster yang merupakan pakar dalam bidang
masing-masing. Sehingga kini, sebanyak 123 kajian R&D dilaksana melibatkan
pelbagai institusi yang merangkumi empat klaster di atas. Sebahagian besar
kajian tersebut telah menghasilkan Standard of Procedure yang boleh dijadikan
rujukan kepada pengkaji-pengkaji dan pengusaha-pengusaha lain dalam
penanaman herba. Selain daripada itu, pelbagai prototaip-prototaip dihasilkan
dan sebahagiannya telah dipatenkan untuk disasarkan kepada syarikat-syarikat
peneraju yang berminat dalam penghasilan produk herba.
Proses penilaian untuk mencari syarikat-syarikat peneraju dalam
meneraju pasaran produk-produk herba bernilai tinggi juga merupakan salah
10
satu inisiatif utama HDD. Sebanyak 14 syarikat telah berjaya melalui tatacara
penilaian yang ketat untuk dipilih sebagai syarikat peneraju. Syarikat-syarikat
tersebut akan menggunakan geran yang diperuntukkan untuk melaksanakan
kajian pra-klinikal dan klinikal bagi produk yang terpilih yang dikeluarkan oleh
syarikat di makmal-makmal di dalam dan di luar negara. Produk nutraseutikal
bernilai tinggi ini dengan kenyataan-kenyataan kesihatan yang telah disahkan
dengan hasil ujian, akan dipasarkan dan dilancarkan dalam pasaran global yang
dijangka berpotensi untuk dijual pada harga 10 kali ganda daripada harga asal.
Geran Projek Khas
Geran kategori ini disediakan bagi melaksanakan projek yang boleh membantu
merancakkan lagi pembangunan industri herba negara. Beberapa projek khas
yang telah dilaksanakan dengan kerjasama beberapa institusi/agensi antaranya
adalah Herbal Extraction Centre oleh Institute of Bio product Development
(IBD), Herbal Trading House oleh Marditech dan Monograph yang dilaksanakan
oleh Institut Penyelidikan Perubatan (IMR). Monograph merupakan laporan
ilmiah berkaitan herba tempatan yang digunakan sebagai bahan rujukan
penerbitan penting mengenai kualiti pokok herba yang terdapat di Negara.
Website Globinmed.com merupakan aplikasi digital yang menjadikan tempat
rujukan monograf ini. Sepanjang tempoh pelaksanaan projek monograph di
bawah geran ini sebanyak 27 tanaman herba dikenal pasti iaitu merungai,
lempoyang, gelenggang, belalai gajah, halia bara, ketum, cengkih, kayu manis,
cekur, karas, kari, kunyit, lada hitam, cucur atap, peria katak, capa, temu hitam,
lengkuas, bongelai, lemuni, pandan, kantan, limau purut, pecah beling, serai
makan, kenanga dan buah pala.
Selain daripada itu, HDD juga telah menyalurkan geran kepada pihak
FRIM untuk melaksanakan Projek Pengetahuan Tradisi Melayu (TK Malayu) dan
Kajian Rantaian Nilai Industri Herba Terpilih di Semenanjung Malaysia yang
merupakan pangkalan data industri herba di peringkat huluan dan hiliran yang
bertujuan bagi mengenal pasti permintaan dan penawaran bahan mentah dan
penghasilan produk herba. Sistem MyTKDL yang dibangunkan oleh MYIPO
merupakan pangkalan data pengetahuan tradisi dan sumber genetik dari
Malaysia dalam bentuk digital untuk kegunaan semasa meluluskan
permohonan antara geran di bawah projek khas ini.
Pengkomersialan Produk Herba
Pengkomersialan produk herba bernilai tinggi merupakan penunjuk prestasi
utama (KPI) utama dalam pelaksanaan ETP NKEA. NKEA EPP1- Produk Herba
Bernilai Tinggi berusaha untuk mengubah ekonomi ke arah berdaya saing yang
bukan sahaja memberi tumpuan kepada projek-projek herba di luar bandar
11
dan juga ianya dibawa ke peringkat antarabangsa. Di peringkat hiliran, HDD
telah merangka halatuju yang lebih progresif bagi memastikan produk-produk
herba yang dihasilkan memberi pulangan berganda kepada negara.
Pelbagai inisiatif pengkomersialan diselaras dan dilaksanakan samada
di peringkat tempatan dan luar negara oleh HDD. Penglibatan secara langsung
di dalam pameran dan expo sedang giat dilaksanakan bagi memastikan HDD
menjadi salah satu badan utama Kerajaan dalam menggerak industri herba
tempatan. Antara penglibatan utama HDD di peringkat antarabangsa adalah
melalui pameran herba di Menope, Dubai dan Anaheim, Amerika Syarikat.
Objektif utama penglibatan pameran di peringkat antarabangsa ini adalah bagi
membuka peluang kepada syarikat-syarikat peneraju herba tempatan untuk
bekerjasama dengan syarikat-syarikat luar dalam memasarkan produk herba
mereka. Ini secara langsung bagi mencapai sasaran GNI sebanyak RM2.2 bilion
sebagaimana yang ditetapkan oleh Kerajaan dalam pasaran industri herba.
Penglibatan HDD dalam pameran dan expo di peringkat tempatan sama ada
pameran yang diadakan secara bertema iaitu bulan herba, persidangan dan
konferens berkaitan herba juga secara langsung mengetengahkan produkproduk herba ini supaya lebih dikenali dari segi khasiat penggunaanya oleh
masyarakat.
Selain memasarkan produk-produk herba di outlet-outlet biasa, HDD
juga berhasrat memperluaskan ke peringkat yang lebih premium. Usahasama
dengan pihak berkepentingan dalam membawa produk-produk ini ke tahap
lebih tinggi diatur dan HDD berperanan sebagai pemudah cara untuk syarikatsyarikat herba memasarkan produk mereka. Jalinan perkongsian bijak dan
strategic alliance di antara kerajaan dengan sektor korporat, pertubuhan bukan
kerajaan (NGO) adalah amat penting dalam mempromosikan industri herba ini
di mata dunia. Kerajaan tidak mampu melaksanakan secara bersendirian dan
sentiasa mengharapkan jalinan kerjasama secara holistik daripada semua
pihak. Adalah diharapkan hubungan atau perkongsian strategik yang terjalin ini
dapat dimanfaatkan kepada lebih ramai lagi kumpulan di masa-masa hadapan.
Inisiatif-Inisiatif dan aktiviti sepanjang tahun yang telah dilaksanakan oleh HDD
kelihatan berupaya memacu industri herba negara ke arah yang lebih
sistematik. Peranan HDD dilihat akan terus signifikan sebagai penyelaras
aktiviti di sepanjang rantaian nilai industri herba pada masa hadapan.
RUJUKAN
UNIT PENGURUSAN PRESTASI DAN PELAKSANAAN (PEMANDU). Aras 3 Blok
Timur, Bangunan Perdana Putra, JPM.
Global Industry Analyst Inc, US. http://www.strategyr.com
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PERANAN FRIM DALAM MEMPERKASA INDUSTRI HERBA DI MALAYSIA
MA Rasadah
Institut Penyeldikan Perhutanan Malaysia (FRIM), 52109 Kepong, Selangor
Tel: 03-6279 7330 E-mel: [email protected]
ABSTRAK
Bahagian Hasilan Semula Jadi (BHS), FRIM telah ditubuhkan pada tahun 1995
telah diberi mandat secara langsung untuk melaksanakan beberapa projek
penyelidikan yang berteraskan penemuan sebatian semula jadi (Natural
Products Discovery) serta pembangunan produk herba & penjagaan kesihatan
daripada tumbuhan ubatan dan beraroma yang berorientasikan pasaran.
Terdapat dua pendekatan yang dilaksanakan oleh Bahagian Hasilan Semula Jadi
iaitu program jangka pendek (1-3 tahun) dan jangka panjang (3-5 tahun).
Program jangka pendek adalah untuk penghasilan data-data saintifik yang
diperlukan bagi menyokong dakwaan dan kegunaan herba untuk kesihatan &
penjagaan diri, sementara jangka panjang pula adalah penghasilan prototaip
produk dan proses komersialisasi. Penyelidikan yang terperinci adalah
diperlukan untuk menghasilkan produk yang berkualiti, berkesan dan selamat
digunakan. Sehingga kini lebih daripada 30 prototiap produk herba telah
dapat dihasilkan.
Berikut adalah usaha-usaha yang sedang dijalankan oleh FRIM dalam usaha
untuk memantapkan industri herba ke peringkat yang boleh dibanggakan:
 Domestikasi dan penanaman tumbuhan herba
 Bioprospek sebatian semula jadi dari sumber hutan
 Kawalan kualiti produk herba
 Formulasi dan pembangunan produk
 Teknologi lepas tuai dan pemprosesan
 Penemuan sebatian perintis dari sumber semula jadi
FRIM juga terlibat dengan pelbagai program dan latihan teknologi herba
dengan pelbagai kementerian seperti dengan Kementerian Luar Bandar dan
Wilayah (KKLW), MOA, KKM, MARA, MTDC dan MITI untuk melatih usahawan
herba dalam usaha untuk meningkatkan mutu penghasilan produk herba yang
mampu menembusi pasaran antarabangsa. Selain dari melaksanakan
penyelidikan, BHS juga menawarkan perkhidmatan dan berkongsi teknologi
terkini dalam pemprosesan & penghasilan produk herba. Sehingga kini lebih
daripada 200 usahawan herba telah mendapat manafaat dari program dan
latihan yang dikendalikan.
13
Kata kunci: FRIM, Bahagian Hasilan Semula Jadi, perintis, peranan, industri
herba
LATAR BELAKANG
Langkah baru dunia dalam era ekonomi global menuntut satu transformasi
pembangunan industri herba yang berteknologi tinggi di negara kita, Malaysia.
Penggunaan tumbuhan ubatan dan herba dalam perubatan tradisonal telah
melalui pelbagai transformasi sejak beberapa dekad yang lalu. Ianya dapat
menjana ekonomi negara sekiranya dieksplotasi dengan baik tanpa
menjejaskan eko-sistem terutamanya hutan semula jadi. Untuk itu, potensi
tumbuhan ubatan dilihat begitu besar sehingga dijadikan sebagai projek
pemulaan yang pertama (EPP1) dalam Bidang Keberhasilan Utama Ekonomi
(NKEA) pertanian di bawah seliaan Kementerian Pertanian dan Industri Asas
Tani Malaysia (MOA). Tumbuhan ubatan kini telah dinilai daripada segi aspek
farmaseutikal dan klinikal serta permintaan pasaran global yang meluas telah
meningkatkan nilai ekonominya setaraf dengan ubatan moden. EPP1 ini
memfokuskan ke arah memperkasakan kualiti produk dan usaha mempromosi
untuk menembusi pasaran global untuk produk nutraseutikal dan ubatan
botanikal.
Permintaan terhadap tumbuhan ubatan dan herba telah melonjak
semenjak kehadiran pelbagai produk baru seperti herba nutraseutikal,
makanan kesihatan, herba kosmeseutikal dan produk penjagaan diri yang
berasaskan tumbuhan herba di pasaran. Secara global, penggunaannya
meningkat dengan kadar pertumbuhan 10-20% setahun. Pasaran dunia
keseluruhan untuk tumbuhan ubatan dan produk herba telah dianggarkan
sekitar US$ 60 billion pada tahun 2000 dan dijangka mencecah US$ 5 trillion
menjelang tahun 2050. Di Malaysia, nilai pasaran herba dijangka meningkat di
antara 15-20% setahun dan diunjurkan mencapai US$ 9.4 billion pada tahun
2020. Disebabkan pengunaan produk herba bersifat global dan diamalkan oleh
hampir 80% penduduk dunia serta kemunculan penyakit-penyakit baru,
penyelidikan dan pembangunan terhadap tumbuhan ubatan masih diteruskan
di seluruh dunia bagi menghasilkan produk yang berkualiti dan berkesan serta
penemuan ubat –ubat baru untuk manusia sejagat.
14
PROGRAM-PROGRAM PENYELIDIKAN
Penyelidikan terhadap tumbuhan ubatan telah bermula di FRIM semenjak
tahun 90-an. Teras penyelidikan ini terus berkembang dan bertambah mantap
dengan penubuhan Bahagian Tumbuhan Ubatan pada Januari 1995 yang
kemudiannya dikenali sebagai Bahagian Hasilan Semula Jadi pada Februari
2011 untuk memberti fokus kepada penyelidikan dan pembangunan teknologi
dalam bidang herba dan tumbuhan ubatan.
Bahagian ini telah menggunakan pendekatan pelbagai disiplin dalam
melaksanakan penyelidikan untuk membangunkan ekstrak piawai dan produk
protaip daripada tumbuhan ubatan dan beraroma (MAPs) seperti kosmetik,
nutraseutikal, makanan berfungsi dan fitoperubatan (phytomedicine). Pada
masa yang sama, bahagian telah mewujudkan strategi pelaksanaan pelan
tindakan jangka pendek yang tertumpu kepada pembangunan produk herba
manakala pelan tindakan jangka panjang adalah lebih kepada penemuan
sebatian bioaktif sebagai bahan perintis bagi memacu pembangunan ubatan
farmaseutikal. Bahagian ini mempunyai fasiliti yang lengkap, daripada ladang
herba contoh yang bertaraf Skim Organik Malaysia (SOM) serta perpustakaan
ekstrak, komponen kimia dan minyak pati sehinggalah pusat pemprosesan
herba bertaraf Amalan Pengilangan Baik (GMP).
Empat program yang dikhususkan untuk menjalankan penyelidikan di
bahagian ini secara holistik ialah Program Biosumber, Fitokimia, Bioaktiviti dan
Program Pembangunan Produk. Program-program ini memfokus kepada
bidang-bidang keutamaan seperti di bawah dalam menghasilkan produk herba
yang berkualiti, berkesan, dan selamat digunakan.
(i) Domestikasi dan penanaman tumbuhan herba
Aktiviti merangkumi pengecaman identiti MAPs, dokumentasi maklumat
etnoperubatan, pemuliharaan ex situ MAPs dan penanaman herba mengikut
Skim Organik Malaysia (SOM).
(ii) Bioprospek sebatian semula jadi daripada sumber hutan.
Aktiviti penyelidikan meliputi fitokimia, pemprofilan komponen/fraksi kimia
dan bioaktif serta penilaian terapeutik untuk mengesahkan kenyataan tradisi
dan meninjau potensi-potensi baru.
(iii) Kawalan kualiti produk herba
Jaminan kualiti merupakan komponen yang penting dalam pembangunan
produk herba. Dengan adanya peralatan yang baharu, pendekatan baharu
dalam jaminan kualiti telah ditemui. Ini termasuk penggunaan FT-IR, HS-SPMEGCMS, HPLC, LCMS dan elektronik nose dalam membina profil capjari bagi
15
spesies tumbuhan yang terpilih. Pemiawaian memainkan peranan yang penting
dalam penambah nilai. Pemiawaian ekstrak menjadi norma bagi produk yang
berkualiti.
(iv) Formulasi dan pembangunan produk
Kajian penyelidikan dalam bidang ini menjurus kepada memformulasikan
produk herba, nutraseutikal dan kosmeseutikal yang berkualiti dan berdaya
saing daripada sumber tumbuhan ubatan.
(v) Teknologi lepas tuai dan pemprosesan
Kajian penyelidikan dalam bidang ini menjurus kepada pembangunan dan
pemprosesan produk herba yang berpiawai dan selamat digunakan.
(vi) Penemuan sebatian perintis daripada sumber semula jadi
Kajian penyelidikan ini menjurus kepada mencari sumber ubat-ubatan yang
berpotensi untuk dibangunkan. Selain daripada itu, makmal ini juga
berkeupayaan menyimpan kultur hidup dan mewujudkan pangkalan data
sumber genetik mikrob aktinobakteria untuk digunakan dalam bioteknologi &
biofarmaseutikal.
Selain daripada empat program di atas, penglibatan aktif daripada
Bahagian Bioteknologi Perhutanan (Cawangan Baik Biak Herba, Genetik &
Kultur Tisu) dan Bahagian Inovasi dan Komersialisasi (Pusat Teknologi Herba)
menjadikan FRIM sebagai agensi penyelidikan herba dan tumbuhan ubatan
yang unggul di Malaysia.
KEJAYAAN DAN PENCAPAIAN
1. Pusat Autentikasi herba
Pusat ini berperanan dalam membangun dan menawarkan skim pensijilan
herba daripada segi aspek pengesahihan bahan mentah ke arah pemantapan
industri herba Malaysia dengan mempertingkatkan kualiti, keselamatan dan
efikasi produk yang dihasilkan. Pusat ini juga berperanan penting sebagai
makmal pengesahihan herba untuk Perkhidmatan Kuarantin dan Pemeriksaan
Malaysia (MAQIS) di bawah MOA serta menyumbang kepada pembangunan
sektor herba (EPP1) di bawah NKEA Malaysia.
2. Pemiawaian herba
Ekstrak piawai memainkan peranan penting dalam mempengaruhi penerimaan
pengguna terhadap produk berasaskan herba. Walau bagaimanapun, kajian ke
atas aspek pemiawaian bagi pemprosesan dan pengeluaran ekstrak herba di
Malaysia masih diperingkat awal. Untuk itu pihak FRIM mengorak langkah
16
dalam menjalankan kajian ini. Terkini lebih daripada 35 jenis ekstrak piawai
yang terdiri dari metabolit primer dan sekunder telah dapat dihasilkan. Selain
daripada itu, lebih daripada 12 ekstrak piawai untuk aktiviti antioksidan telah
dibangunkan dan dilesenkan kepada pihak industri herba. Beberapa Prosedur
Operasi Piawai (SOP) yang mematuhi garis panduan perundangan pasaran
telah dibangunkan dan sedia untuk dikomersialkan. Antaranya ialah mengkudu,
kacip fatimah, cucur atap, gelenggang, misai kucing, karas, pokok kapalterbang,
minyak pati dan sebagainya.
3. Polisakarida
FRIM merupakan satu-satunya badan penyelidikan di Malaysia yang
menjalankan R&D secara intensif terhadap polisakarida daripada tumbuhan
ubatan sebagai sumber produk farmaseutikal dan makanan berfungsi. Hasil
penyelidikan polisakarida daripada buah mengkudu telah dipaten dan
dikomersialkan kepada Syarikat Poly-Xtract Sdn. Bhd. untuk menghasilkan
fraksi polisakarida pada skala pandu (pilot scale) dan fraksi polisakarida ini
telah digunakan oleh Syarikat Nourish Care Sdn. Bhd. sebagai bahan aktif untuk
penghasilan produk makanan jenama Le’Natura iaitu biskut dan kopi putih
mesra diabetes. Produk-produk ini telah pun berada di rangkaian-rangkaian
pasar raya seperti Aeon Jusco, OTK, Mydin, Tesco dan Giant. Hasil penyelidikan
ini juga telah dipilih di antara 42 projek penyelidikan daripada seluruh negara
oleh pihak Agensi Inovasi Malaysia (AIM) sebagai peluang perniagaan inovatif
(IBO) yang berpotensi menjana pendapatan RM 1 billion selepas 3 tahun.
Inovasi daripada projek penyelidikan ini juga telah terpilih ke peringkat akhir
Anugerah Inovasi Negara 2012.
4. Pengredan gaharu
Pemintaan terhadap kayu gaharu dan hasilan produk berasaskan gaharu sangat
tinggi. Penentuan harga dan gred bagi kayu dan minyak gaharu adalah tidak
seragam. Untuk itu, FRIM telah memperkenalkan satu sistem pengklasifikasi
pintas (gaharu sense) yang berkonsepkan teknologi alat pengesanan bauan
(hidung elektronik). Sistem pengredan ini telah berjaya dipatenkan dan sedia
untuk ditawarkan kepada industri gaharu di negara ini.
5. Antioksidan/pemutih kulit dan Asai Cap-E
Penyelidikan antioksidan/pemutih kulit (skin whitening) di FRIM telah
dijalankan semenjak tahun 1995. Antioksidan sering mendapat sambutan
dalam penjagaan kesihatan terutama dalam mencegah penyakit kronik
degeneratif dan melambatkan tanda-tanda penuaan. Walau bagaimanapun,
kebelakangan ini pengujian keberkesanan antioksidan bagi hasilan semula jadi
dipersoalkan. Adakah pengambilan antioksidan meresap masuk ke dalam
sistem badan, salur darah, atau bolehkah ianya memberi perlindungan kepada
kerosakan akibat kesan oksidatif? Untuk itu, kumpulan penyelidik FRIM telah
17
membangunkan asai Cap-E untuk menganalisa kebolehdapatan bio antioksidan
pada peringkat sel dalam hasilan semula jadi dan ekstrak tumbuhan. Ujian
antioksidan/pemutih kulit dan Cap-E telah ditawarkan kepada pihak industri
herba.
6. Konservasi tumbuhan ubatan/Perpustakaan
Setakat ini, Bahagian telah berjaya mengumpul spesimen rujukan untuk 1,147
spesies MAPs. FRIM juga telah mewujudkan konservasi ex situ MAPs dengan
penanaman lebih daripada 200 spesies di Taman Ethnobotani di Kepong.
FRIM telah berjaya mengumpulkan 1,078 fitoekstrak, 35 ekstrak
piawai, 150 minyak pati, 135 sebatian fitokimia dan lebih daripada 12,000
kultur aktinobakteria dalam perpustakaan hasilan semula jadinya. Kejayaan ini
sangat penting bagi program pembangunan dalam penemuan sebatian bioaktif
bernilai terapeutik. Simpanan atau repositori serta pangkalan data yang
sistematik dan komprehensif ini akan membantu dalam penggunaan sumber
biodiversiti negara secara optima dan mampan dalam usaha menjana ekonomi
baru negara.
Pada masa yang sama, Bahagian Hasilan Semula Jadi juga menjalankan
penilaian bioefikasi dan potensi bio dan mewujudkan 25 bioasai berkaitan
dengan bioterapeutik, kegagalan metabolik dan sistem pengenalpastian
mikrobial. Buat masa ini , Bahagian mempunyai pangkalan data bagi 564
spesies bakteria aerobik gram-negatif, 480 spesies bakteria anerobik, 619
spesies filamen kulat dan 267 species yis serta 19 Methicilin superbug
Staphyloccocus aureus untuk penyelidikan.
7. Pengiktirafan
Kejayaan Bahagian Hasilan Semula Jadi dalam tempoh yang singkat iaitu sejak
tahun 1995 dengan kepakaran 50 pegawai penyelidik dalam menjalankan
kajian herba dan hasilan semula jadi banyak dibantu oleh kemudahan makmal
berteknologi tinggi. Pusat Teknologi Herba (HTC) yang dimiliki FRIM, dilengkapi
dengan peralatan sejajar dengan matlamat dan peranannya sebagai pentas
yang menghubungkan agensi-agensi kerajaan termasuk agensi penyelidikan &
penguatkuasaan dengan pengusaha herba bagi penghasilan produk yang
berkualiti tinggi. Di samping itu, HTC telah menerima persijilan GMP untuk
pemprosesan ubatan berbentuk kapsul dan tablet daripada Kementerian
Kesihatan Malaysia (KKM) pada tahun 2007.
Stesen Penyelidikan di Maran, Pahang (SP Maran) telah dianugerah
SOM pada tahun 2010 untuk tanaman patawali. Tanaman lain yang ditanam di
SP Maran adalah tongkat ali, misai kucing, mengkudu, kacip fatimah, tutup
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bumi, kunyit, limau nipis, serai wangi, lengundi, kalampayan, asam gelugor, dan
karas.
Pada 26 Julai 2011, MOSTI telah mengiktirafkan FRIM sebagai Pusat
Kecemerlangan dalam Pembangunan Hasilan Semula Jadi (BNP Centre of
Excellence) di Malaysia. Ini sejajar dengan keupayaan FRIM dalam
membangunkan enam makmal berstatus BioNexus iaitu Makmal Fitokimia,
Farmakognosi, Formulasi Produk dan Pemprosesan Herba daripada Bahagian
Hasilan Semula Jadi serta Makmal Genetik dan Tisu Kultur daripada Bahagian
Bioteknologi Perhutanan yang berperanan sebagai pusat kecemerlangan dalam
memberi perkhidmatan teknikal serta membangunkan produk berasaskan
herba
Sejak tahun 1995, sebanyak 30 anugerah inovasi telah diperolehi oleh
bahagian ini di peringkat kebangsaan dan antarabangsa. Anugerah Geneva
Awards, Innova Brussel, Anugerah Saintis Muda Merck serta Anugerah
Pencapaian Saintifik IUFRO adalah antara lapan pengiktirafan antarabangsa
kepada sumbangan dan hasil R&D Bahagian Hasilan Semula Jadi FRIM;
termasuk 18 Anugerah Inovasi peringkat kebangsaan yang diterima sejak 2006.
Bahagian ini juga telah menghasilkan 30 prototaip produk herba; 15
daripadanya telah berjaya dikomersialkan manakala empat produk lagi akan
dikomersialkan tidak lama lagi; menghasilkan 5 paten, 8 pemfailan paten serta
lebih daripada 49 pendedahan reka cipta (Invention Disclosure). Selain itu,
secara purata empat penerbitan, 20 artikel serta 140 kertas kerja diterbitkan
dalam jurnal berimpak tinggi setiap tahun bagi menyebarkan maklumat serta
kajian R&D FRIM kepada pihak awam dan golongan penyelidik.
8. Pemindahan teknologi
Setiap tahun, bahagian ini telah melatih lebih daripada 250 orang usahawan
herba dan pemakanan melalui khidmat nasihat dan pemindahan teknologi
serta menawarkan antara 100 hingga 150 perkhidmatan pemprosesan herba.
Kebanyakan program latihan yang dianjurkan setiap tahun membantu industri
herba dalam penghasilan produk yang berkualiti, berkesan dan selamat
digunakan. Program-program ini melibatkan usahawan dari MARA, KKLW,
MTDC dalam usaha untuk meningkatkan mutu penghasilan produk herba dan
penanaman tumbuhan herba yang berkualiti.
Program latihan siswazah (FMBioSIS) berjaya melatih siswazah menjadi
usahawan yang berjaya dalam bidang bioteknologi dan herba. Sebanyak 10
syarikat yang ditubuhkan bersama Perbadanan Pembangunan Teknologi
Malaysia (MTDC) akan mengkomersialkan hasil R&D FRIM. Dalam program ini,
Bahagian telah berupaya melatih empat syarikat dalam pembangunan produk
19
herba dan spa. Program ini boleh menjadi contoh kepada institusi-institusi
pengajian tinggi tempatan untuk memulakan langkah yang sama.
KESIMPULAN
Kejayaan dan potensi masa hadapan industri herba di Malaysia amat
bergantung kepada keupayaan kita untuk mewujudkan piawaian atau
standard dan garis panduan yang boleh digunakan oleh pengeluar, pengilang
dan pemasar produk herba dan secara langsung akan mematuhi garispanduan
dan peraturan yang telah ditetapkan oleh badan antarabangsa seperti
Pentadbiran Makanan dan Ubat-ubatan (FDA) dan Kesatuan Eropah (EU). Ini
juga perlu seiring dengan strategi pembentukan komuniti ekonomi Asean (AEC)
yang diwujudkan pada 2015 supaya lebih kompetitif di pasaran tunggal yang
lebih terbuka. Kepakaran dan fasiliti yang wujud di FRIM mampu melonjakkan
industri herba negara ke tahap yang boleh dibanggakan. Malah terdapat
usahawan hasil didikan FRIM telah menjulang nama FRIM di persada dunia
melalui pengeluaran bahan mentah atau produk yang boleh diketengahkan dan
berupaya meraih pendapatan sebanyak RM6 hingga RM7 juta setahun.
Ringkasnya, Bahagian Hasilan Semula Jadi di FRIM adalah perintis kepada usaha
memanfaatkan tumbuhan herba sesuai dengan peranannya membangunkan
sumber semula jadi hutan.
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MENYUSUN STRATEGI BERPANDUKAN FAKTA RANTAIAN NILAI
INDUSTRI HERBA
M Marzalina, AB Ariff Fahmi, AR Rohana & R Rosniza
Tel: 03-6279 7215 Faks: 03-6273 6587 E-mel: [email protected]
ABSTRAK
Industri herba Malaysia merupakan satu bidang yang telah dikenalpasti
berpotensi sebagai sumber kekayaan baharu negara. Dasar Agromakanan
Negara (DAN) 2011 - 2020 serta Bidang Ekonomi Utama Negara (NKEA) sektor
pertanian jelas menekankan pembangunan industri herba sebagai salah satu
agenda penting yang perlu bergerak secara efektif. Pemahaman terhadap
struktur rantaian nilai industri herba serta pihak yang terlibat secara langsung
di dalam industri merupakan asas terpenting dalam merangka strategi yang
berkesan dalam memacu keseluruhan industri ke satu peringkat yang lebih
tinggi. Melalui Kajian Rantaian Nilai Industri Herba Terpilih di Semenanjung
Malaysia yang dijalankan oleh Institut Penyelidikan Perhutanan Malaysia
(FRIM), pertemuan secara individu mahupun melalui bengkel konsultasi
dengan semua pihak berkepentingan dalam industri herba telah diadakan.
Dikaitkan dengan fakta yang diperolehi, kertas kerja ini akan dapat
dimanfaatkan bagi membantu pembentukan strategi yang terfokus untuk
memperkasa industri herba tempatan. Tiga spesies herba akan diketengahkan
bagi merealisasi usaha menjadikan hub herba Malaysia dengan identiti
tersendiri. Pembentukan pelan pembangunan strategik yang komprehensif
merangkumi komponen penyelidikan dan pembangunan (R&D), pembangunan
modal insan, struktur bantuan kewangan industri, bahan mentah, struktur
perniagaan dan pelan pemasaran industri merupakan agenda penting yang
perlu digerakkan sehaluan secara bersama. Gabungan gagasan kerjasama
semua pihak sangat perlu sekiranya semua serius ke peringkat global untuk
meningkatkan ekonomi dan kesihatan rakyat.
Kata kunci: strategi, rantaian nilai, industri herba, hab herba Malaysia
PENGENALAN
Industri herba secara umumnya merupakan suatu bidang yang mempunyai
potensi besar untuk diterokai terutamanya bagi negara seperti Malaysia.
Menurut statistik WHO (2013), permintaan di China meningkat sebanyak 20%
dalam tempoh setahun menjanakan US$83.1 billion, sementara peningkatan
sebanyak 68% di Korea telah menjanakan US$7.4 billion dalam tempoh 5
tahun. Maka ini jelas menunjukkan potensi industri herba sebagai sumber
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kekayaan baharu negara sekiranya negara kita benar-benar serius untuk
menjadikan Malaysia sebagai salah satu hab herba dunia.
Dasar Agromakanan Negara (DAN) mengunjurkan pengeluaran herba
Malaysia bakal mencapai 73,000 tan metrik pada tahun 2020 berbanding
17,000 tan metrik pada tahun 2010. Jumlah keluasan tanaman herba turut
diunjurkan meningkat sebanyak 15% setahun daripada 1,000 hektar pada 2010
kepada 4,000 hektar pada tahun 2020.
Bidang Ekonomi Utama Negara (NKEA) diperkenalkan di bawah
Program Transformasi Ekonomi (ETP) merangkumi 12 sektor di mana bidang
pertanian merupakan salah satu daripada sektor yang diberi perhatian. Di
bawah NKEA pertanian pula, sebanyak 17 Projek Penggerak Ekonomi (EPP)
telah dikenalpasti dimana bidang industri herba terlibat secara langsung di
bawah EPP 1 iaitu unlocking value from Malaysia’s biodiversity through highvalue herbal product.
Rantaian Nilai Industri Herba (RNIH) merangkumi pembekal bahan
tanaman, penanam, pemproses/pengeluar produk herba, pemborong, peruncit
dan juga pengguna. FRIM telah menjalankan beberapa kajian melibatkan
kesemua peringkat bagi memahami struktur rantaian nilai industri herba secara
keseluruhannya. Objektif kertas kerja ini ialah membentuk strategi yang
terfokus dalam usaha memperkasa industri herba tempatan dan seterusnya
mengukuhkan usaha pewujudan hab herba Malaysia dengan identiti tersendiri.
KAEDAH
Kertas kerja ini dibangunkan berdasar input bancian, survei dan siri bengkel
yang dijalankan oleh FRIM melalui kajian rantaian nilai industri herba terpilih di
Semenanjung Malaysia. Temu bual melibatkan pelbagai pihak berkepentingan
serta pemerhatian kumpulan penyelidikan terhadap senario semasa industri
herba dan rujukan pelbagai sumber turut dilakukan.
PENEMUAN DAN PERBINCANGAN
Hasil bancian Kajian Rantaian Nilai Industri Herba Terpilih Semenanjung
Malaysia, menunjukkan terdapat perbezaan antara spesies herba ditanam dan
diproses oleh pengusaha. Rajah 1 menunjukkan senarai bilangan penanam 10
spesies terbanyak serta keluasan ladang berbanding bilangan pengusaha dalam
pemprosesan berdasarkan siri bancian yang dijalankan pada tahun 2014 –
2015. Keadaan ini memberi gambaran awal terhadap kepincangan koordinasi
antara permintaan dan penawaran dalam rantaian. Penentuan nilai
22
komersialisasi spesies turut penting bagi menentukan hab herba Malaysia
sentiasa di tahap optimum.
Rajah 1. 10 spesies yang paling banyak ditanam berbanding 10 spesies yang
paling banyak digunakan dalam pemprosesan/pengusaha.
Rohana et al. (2015b) turut membincangkan mengenai keperluan
untuk industri herba mengurangkan kebergantungan terhadap import bahan
mentah dan sebaliknya memperkasakan pihak pengeluar bahan tanaman
(tapak semaian) dan peladang itu sendiri bagi memenuhi keperluan
pemproses/pengusaha produk herba. Perkara ini berkait rapat dengan
permintaan orang ramai. Oleh itu pengguna perlu mendapat maklumat yang
tepat melalui strategi promosi yang berkesan.
Walau bagaimanapun, dari input Bengkel Konsultasi Pihak
Berkepentingan Industri Herba Semenanjung Malaysia (Februari 2015), majoriti
pihak berpandangan industri herba kita mempunyai beberapa kekuatan untuk
dieksploitasi terutamanya berkaitan sumber, kepakaran dan teknologi. Ini turut
diutarakan oleh mereka yang kami temu bual. Strategi yang baik perlu
mengambilkira faktor-faktor halangan dan sumber semasa yang wujud.
Selain itu, suatu pelan strategik yang memfokuskan matlamat utama untuk
pembangunan hab herba Malaysia beridentiti tersendiri perlu dirangka segera.
Kami mencadangkan pemilihan spesies berpotensi untuk dikembangkan adalah
berdasarkan beberapa kriteria seperti berikut:
1. Spesies yang mempunyai maklumat lengkap kajian dan rantaian
industri berserta kelebihan mampu bersaing di peringkat global contoh: Tongkat ali.
23
2. Spesies yang digunakan secara tradisional, disokong secara saintifik
serta mempunyai potensi untuk bersaing di peringkat ASEAN - contoh:
Kacip fatimah.
3. Spesies yang turut dikaji serta popular disebabkan trend/keperluan
semasa - contoh: Belalai gajah
Daripada tiga spesies tersebut, pelan pembangunan strategik
hendaklah merangkumi komponen penyelidikan dan pembangunan (R&D),
pembangunan modal insan, struktur bantuan kewangan industri, bahan
mentah, struktur perniagaan dan pelan pemasaran industri perlu dibangunkan.
Pelan ini bakal merupakan agenda penting yang perlu digerakkan sehaluan
secara bersama ke arah merealisasikan hab herba Malaysia tersebut.
Melalui pelan pembangunan strategik yang dibangunkan, koordinasi
antara setiap peringkat rantaian serta pemain utama perlu dikenal pasti. Pihakpihak berkuasa dan intelektual yang mempunyai kekuatan di setiap peringkat
rantaian hendaklah bersama-sama memberi input, bekerjasama dan
menyokong kepada hab herba Malaysia. Antara lain seperti mengukuhkan
aspek penyelidikan, keberkesanan, kualiti dan keselamatan herba;
memperkenalkan bahan tanaman elit spesies terpilih untuk ditanam;
menyediakan teknologi termoden bagi menghasilkan bahan mentah;
menghubungkaitkan situasi permintaan dan penawaran; menentukan harga
terbaik pembelian; meningkatkan usaha promosi dalam menjenamakan herba
Malaysia di persada dunia serta mencipta lebih banyak peluang pasaran (blue
ocean strategy) agar permintaan sentiasa wujud tanpa henti.
RUMUSAN
Berasaskan fakta dan keperluan semasa, pelan pembangunan strategik yang
dicadangkan merupakan suatu garis panduan yang dapat mengukuhkan hasrat
membina hab herba Malaysia dengan identiti tersendiri. Walau bagaimanapun,
kejayaan sebenar hanya akan mampu dicapai melalui penggubalan polisi
beserta penguatkuasaan yang serius oleh semua pihak sama ada dari dalam
mahupun di luar rantaian industri herba.
RUJUKAN
Gerard, B., Hood, S., Ruzy Suliza, H., Christof, J., Joerg, G., & Zurinawati, Z.A.
(Eds.). (2009). Health and Beauty from the Rainforest ─ Malaysian
Traditions of Ramuan. Kuala Lumpur: Editions Didier Millet Pte Ltd.
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Jabatan Pertanian Semenanjung Malaysia. (2013). Statistik Tanaman (Sub
Sektor Tanaman Makanan). Jabatan Pertanian Semenanjung Malaysia.
Putrajaya: Jabatan Pertanian Semenanjung Malaysia.
Kementerian Pertanian dan Industri Asas Tani. (2011). Dasar Agromakanan
Negara 2011─2020. Putrajaya: Bahagian Perancangan Strategik dan
Antarabangsa, MOA.
Rohana, A. R., Nur Fazreen, Z., Ariff Fahmi, A. B., Nur Syazni, A., Siti Zubaidah,
S., Lim, H. F., et al. (2015). Directory of Herbal Cultivators in Peninsular
Malaysia. Kuala Lumpur: Forest Research Institute Malaysia.
Rohana, A. R., Nur Fazreen, Z., Ariff Fahmi, A. B., Nur Syazni, A., Siti Zubaidah,
S., Lim, H. F., et al. (2015). Value Chain Study for Holistic Development
of Herbal Industry. Proceeding 2015 3rd International Conference on
Rural Development & Entrepreneurship, (pp. 211─219). Hebei Province,
China.
Vimala, S. (Ed.). (2013). Malaysian Herbal Heritage. Kuala Lumpur: Forest
Research Institute Malaysia (FRIM).
World Health Organization. (2013). WHO Traditional Medicine Strategy: 20142023. Hong Kong SAR, China: WHO.
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PENDOKUMENAN PENGETAHUAN TRADISI MELAYU
TUMBUHAN UBATAN DI SEMENANJUNG MALAYSIA
BERKAITAN
M Nik Musaadah, M Dionysia, MS Abdul Hayat, Z Nurul Husna, J Fadzureena,
B Intan Nurulhani, HF Lim, AL Tan, R Rosniza, MN Madihah, MK Noor Aimie,
MS Md Azharulzaman, MA Nor Azah, M Mastura & H Norini
Dengan kerjasama: Jawatankuasa Teknikal Projek Pengetahuan Tradisi Melayu
ABSTRAK
Pengetahuan tradisi Melayu berkaitan tumbuhan ubatan perlu didokumen dan
dipulihara demi menyokong sasaran ke arah memperkasakan industri herba.
Pendokumenan pengetahuan tradisi ini bermula dengan aktiviti bancian
pengamal perubatan tradisional Melayu, survei penggunaan tumbuhan ubatan
dan juga kutipan sampel tumbuhan ubatan atau peralatan. Selain daripada
pengamal, manuskrip perubatan Melayu turut dijadikan sumber rujukan. Hasil
pendokumenan ini didapati masih banyak tumbuhan ubatan digunakan dalam
perubatan tradisional Melayu yang boleh dijadikan asas untuk mengenal pasti
spesies yang berkepentingan ekonomi. Hasil pendokumenan ini telah
dikumpulkan dalam pelbagai bentuk yang boleh dijadikan sumber rujukan
generasi akan datang.
Kata kunci: pengetahuan tradisi, tumbuhan ubatan, Melayu
PENGENALAN
Pengetahuan tradisi termasuk pengetahuan tradisi Melayu berkaitan
tumbuhan ubatan wajar dipulihara kerana ia mempunyai nilai ekonomi selain
daripada menghalang berlakunya bio-piracy dan juga salah guna
(misappropriation). Selaras dengan matlamat Program Transformasi Ekonomi
untuk menjadikan herba sebagai salah satu sumber ekonomi baru negara,
pengetahuan tradisi berkaitan tumbuhan ubatan boleh dijadikan asas untuk
mengenal pasti tumbuhan ubatan yang berpotensi untuk kajian lanjutan dan
diberikan nilai tambah.
Sehubungan itu satu projek bertajuk Pendokumentasian Secara
Komprehensif Pengetahuan Tradisi Melayu Berkaitan Tumbuhan Ubatan di
Semenanjung Malaysia dilaksanakan. Projek ini telah dibiayai oleh Kementerian
26
Pertanian dan Industri Asas Tani (MOA). Pelaksanaan aktiviti projek ini diterajui
oleh Institut Penyelidikan Perhutanan Malaysia (FRIM) dengan kerjasama
Herbwalk Consultancy; Institut Penyelidikan Perubatan (IMR) dan Bahagian
Perubatan Tradisional dan Komplementari, Kementerian Kesihatan Malaysia;
Perbadanan Harta Intelek Malaysia (MyIPO); Perpustakaan Negara Malaysia
dan Dr Harun Mat Piah (pakar filologi).
Objektif utama projek ini adalah untuk mendokumenkan pengetahuan
tradisi Melayu berkaitan penggunaan tumbuhan ubatan daripada pengamal
perubatan tradisional Melayu dan juga beberapa manuskrip perubatan Melayu
terpilih.
KAEDAH
Projek ini melibatkan aktiviti-aktiviti berikut:
i.
Perolehan keizinan (prior informed consent, PIC)
Setiap pengamal yang ditemui akan diberi keterangan berkaitan projek dan
memohon keizinan pengamal untuk berkongsi ilmunya untuk
didokumenkan dalam penerbitan komersial dan bukan komersial.
Pengamal yang memberi persetujuan akan menandatangani surat keizinan
(prior informed consent, PIC). Perolehan keizinan ini dilakukan bagi setiap
aktiviti yang dilaksanakan.
ii. Bancian pengamal perubatan tradisional Melayu di Semenanjung
Malaysia
Pengamal dikenal pasti melalui pengerusi jawatan kuasa kemajuan dan
keselamatan
kampung/penghulu/penggawa/pengerusi
persatuan
penduduk. Bancian dilakukan secara bersemuka dan temu bual separa
berstruktur dilaksanakan.
iii. Soal selidik/survei berkaitan pengunaan tumbuhan ubatan dan sosio
ekonomi ke atas pengamal terpilih
Berdasarkan hasil bancian, pengamal yang terlibat dalam aktiviti ini dikenal
pasti. Temu bual separa berstruktur secara bersemuka telah dilaksanakan.
iv. Kutipan sampel tumbuhan ubatan atau peralatan
Kutipan sampel tumbuhan ubatan atau peralatan telah dilaksanakan
berdasarkan maklumat bancian dan survei. Spesimen herbarium
disediakan dan pengecaman spesies dilaksanakan. Spesimen hidup juga
turut dikutip dan dipulihara di Taman Etnobotani FRIM. Perolehan
peralatan atau penyediaan replika peralatan dilaksanakan oleh IMR.
v. Transliterasi manuskrip perubatan Melayu/kitab tib
27
Transliterasi daripada Jawi klasik kepada Rumi telah dilaksanakan oleh ahli
filologi.
Bancian Pengamal
Hasil daripada bancian yang dijalankan, terdapat sejumlah 4,975 orang
pengamal perubatan tradisional Melayu di Semenanjung Malaysia telah
dikenalpasti dan seramai 2,934 (59%) pengamal telah ditemu bual. Peratusan
ini dicapai disebabkan oleh masa bancian yang terhad, pengamal tidak berada
di rumah, sakit semasa lawatan, tidak dapat dihubungi atau tinggal terlalu jauh
di pendalaman. Selain itu, terdapat juga kes-kes seperti pengamal yang
meninggal dunia sebelum sempat untuk ditemu bual. Faktor-faktor seperti ini
sememangnya tidak dapat dielakkan.
Soal Selidik/Survei Berkaitan Pengunaan Tumbuhan Ubatan dan Sosio
Ekonomi ke atas Pengamal Terpilih
Berdasarkan perkiraan statistik, saiz minimum responden yang diperlukan
untuk kajian survei adalah 341 orang bagi hasil bancian seramai 2,934 orang
responden (Sakeran, 2000). Beberapa kriteria telah ditetapkan untuk pemilihan
341 pengamal tersebut. Hasilnya, seramai 357 pengamal telah ditemu bual,
namun hanya 355 (99%) pengamal telah memberi persetujuan dan menanda
tangani PIC kedua (PIC 2). Matlamat utama soal selidik ini adalah untuk
mendokumenkan maklumat berkaitan tumbuhan ubatan yang digunakan
dalam pengamalan/perubatan tradisional. Memandangkan, dua responden
tidak memberi keizinan, maka analisa data hanya ditumpukan kepada 355
responden.
Dokumentasi Tumbuhan Ubatan
Sebanyak 2,055 tumbuhan ubatan telah berjaya direkod berdasarkan nama
tempatan. Daripada rekod tersebut, sebanyak 431 tumbuhan dikutip dan
dicam ke peringkat spesies manakala sebanyak 461 tumbuhan dapat dikenal
pasti ke taksa tertentu (genus atau famili). Jadi masih terdapat 1163 tumbuhan
yang direkod berdasarkan nama tempatan sahaja. Tumbuhan ini tidak dapat
dikutip atau dikenal pasti spesiesnya kerana tempoh projek yang singkat
menyebabkan aktiviti kutipan sampel untuk penyediaan spesimen herbarium
terhad. Data berkaitan tumbuhan ubatan ini juga diserahkan kepada MyIPO
untuk pangkalan data MyTKDL.
28
Transliterasi Manuskrip Perubatan Melayu/Kitab Tib
Projek ini telah berjaya mentransliterasikan tiga buah kitab tib iaitu MSS 2999,
Kitab Tib Muzium Terengganu dan juga MSS 1292. Hasil transliterasi ini
sebanyak dua buah buku diterbitkan.
Output Projek
Hasil projek ini pada keseluruhannya adalah seperti Rajah 1. Secara ringkasnya
output yang dihasilkan adalah dalam bentuk penerbitan, pangkalan data,
koleksi spesimen baucer/herbarium dan germplasma untuk tujuan konservasi,
dan juga pengumpulan peralatan bagi tujuan penubuhan muzium.
KESIMPULAN
Perubatan tradisional Melayu masih diamalkan dan wajar dipulihara.
Pendokumenan pengetahuan tradisi Melayu adalah satu langkah untuk
mengekal dan memperkasa perubatan tradisional di kalangan masyarakat
Melayu. Pengetahuan tradisi juga adalah asas bagi meningkatkan industri
herba pada masa hadapan dengan diberikan nilai tambah berasas teknologi
dan inovasi terkini.
PENGHARGAAN
Kami sangat berterima kasih kepada semua pengamal yang terlibat dalam
projek ini dan juga pegawai pengumpul data yang telah melaksanakan bancian
dan juga survei. Projek ini dibiayai oleh MOA.
RUJUKAN
Sakeran, U. 2000. Research Method for Business: A Skill Building Approach.
John Wiley & Sons, USA.
29
Kitab Tib/ Manuskrip
Bancian pengamal
perubatan tradisional
Melayu di Sem. Malaysia
& PIC 1
Buku statistik pengamal
Buku rujukan berkaitan
kriteria pengamal (BPTK)
 Buku MSS2999:
pandangan dan
tafsiran perubatan
moden terhadap
manuskrip perubatan
Melayu
 Buku Transliterasi
Kitab Tib Muzium
Terengganu
Koleksi artifak/ peralatan/
manuskrip (IMR)
Survei pengamal terpilih
(penggunaan tumbuhan
ubatan & sosio ekonomi)
& PIC 2
Kutipan sampel &
penyediaan spesimen
herbarium
Pengecaman spesies
tumbuhan
Dokumentasi TK
• Buku spesies di bawah
NKEA
• Buku spesies yang
diguna dalam
perubatan tradisional
Melayu
• Pangkalan data (MyTKDL
& BRAHMS)
Rajah 1. Aktiviti dan output projek secara keseluruhan
30
 Penubuhan Muzium
Perubatan Tradisional
Melayu (IMR)
 Buku Peralatan
Perubatan Tradisional
Buku kajian sosio ekonomi
pengamal
 Koleksi germplasma
tumbuhan ubatan
 Koleksi herbarium
pengetahuan tradisi
Melayu
PERALATAN DALAM PERUBATAN TRADISIONAL MELAYU: KHAZANAH
ILMU DAN PERADABAN WARISAN BANGSA
A Ida Farah, AZ Wan Nurul Syafinaz, Z Nurasyikin & SM Ami Fazlin
Pusat Penyelidikan Perubatan Herba, Institut Penyelidikan Perubatan (IMR),
Jalan Pahang, 50588 Kuala Lumpur
Tel: 03-4041 1662
Faks: 03-4041 1657
ABSTRAK
Masyarakat di Malaysia masih bergantung kepada amalan perubatan
tradisional yang diwarisi sejak turun temurun walaupun ianya tidak lagi diguna
sebagai kaedah utama perawatan dan pencegahan penyakit. Sejarah
penemuan ubatan moden banyak berasaskan pengetahuan tradisi yang melalui
kajian saintifik. Perkongsian pengetahuan dan pendekatan dalam amalan
perubatan tradisional ini mengalami ketirisan akibat ketiadaan sistem
pengumpulan dan penyampaian maklumat secara terurus. Kajian ini merekod
secara terperinci kaedah dalam amalan serta peralatan dan artifak yang
digunakan oleh pengamal dalam menyediakan ubatan dan merawat pesakit.
Maklumat mengenai aAlatan yang dikumpul akan didokumentasikan di dalam
sebuah buku mewah. Alatan-alatan berkaitan pula akan dipamerkan di Muzium
Alatan Perubatan Tradisional Melayu di Institut Penyelidikan Perubatan (IMR).
Pengamal dipilih melalui bancian yang dijalankan melalui projek
“Pendokumentasian Secara Komprehensif Pengetahuan Tradisional Melayu
Berkaitan Tumbuhan Ubatan di Semenanjung Malaysia” berdasarkan jenis
pengetahuan dan peralatan yang digunakan dalam amalan masing-masing.
Temu bual berstruktur menggunakan borang kaji selidik mengumpul maklumat
sejarah dan perincian pengetahuan dan pengalaman mereka. Alatan atau
artifak diperoleh sama ada daripada pengamal semasa temu bual dijalankan
atau dibeli di premis berkaitan. Artifak yang tidak dapat diperolehi dihasilkan
semula melalui kaedah pembuatan replika. Lima amalan utama yang direkod
menerusi kajian yang melibatkan 65 orang responden ini adalah perbidanan
(20 orang), urutan patah dan biasa (14 orang), rawatan kerohanian (13 orang),
penghasilan produk herba (10 orang) dan bomoh patah (7 orang). Sebahagian
daripada pengamal ini menawarkan lebih daripada satu jenis rawatan dalam
perkhidmatan mereka. Koleksi alatan yang terbanyak ialah untuk penyediaan
herba (27 jenis), diikuti rawatan kerohanian (15 jenis), perbidanan (14 jenis),
rawatan umum (14 jenis), urut (6 jenis) dan bomoh patah (2 jenis).
Pendokumentasian pengetahuan ini merekod khazanah ilmu dan amalan
tradisional secara lebih teratur dan mudah diakses oleh masyarakat. Ia juga
menggalakkan generasi muda menghargai dan mengambil peluang
mempelajari kemahiran tradisi di samping mengenali alatan dalam rawatan
tradisional Melayu. Kajian ini menjadi pemangkin dalam memartabat dan
31
memberi penghargaan khusus kepada warisan pengetahuan tradisi Melayu
melalui penggiatan aktiviti pendokumentasian sebelum ia hilang ditelan masa.
Kata kunci: perubatan tradisional, peralatan, kaedah rawatan Melayu,
tumbuhan ubatan, herba
PENGENALAN
Perubatan tradisional atau etnofarmakologi mengetengahkan konsep dan
amalan yang dilazimi oleh masyarakat sesuatu kaum atau bangsa. Sesuatu
amalan itu secara asasnya dipengaruhi budaya masyarakat setempat serta
ketersediaan sumber di persekitaran mereka. Etnofarmakologi ditakrifkan
menurut Kamus Dewan Bahasa dan Pustaka Edisi Keempat (DBP 2013) sebagai
“bidang kajian mengenai tumbuhan yang digunakan untuk merawat penyakit
dan penjagaan kesihatan oleh kumpulan etnik tertentu”. Kajian berkaitan
etnofarmakologi merupakan kajian menyeluruh yang merangkumi aspek
sosiologi, botani, biokimia dan alam sekitar.
Kajian Peralatan Dalam Perubatan Tradisional Melayu ini mensasarkan
pendokumentasian maklumat berkaitan peralatan yang digunakan di dalam
penyediaan ubatan dan perawatan pesakit. Kajian ini didasari daripada Kajian
Pendokumentasian Secara Komprehensif Pengetahuan Tradisional Melayu
Berkaitan Dengan Tumbuhan Ubatan yang diterajui oleh Institut Penyelidikan
Perhutanan Malaysia (FRIM). Kajian yang berlangsung dari 2013 sehingga 2015
ini dibiayai oleh Kementerian Pertanian & Industri Asas Tani (MOA) melalui
projek permulaan Bidang Keberhasilan Utama Ekonomi (NKEA)-Pertanian iaitu
Pembangunan Produk Herba Bernilai Tinggi (EPP1) kelolaan Bahagian
Pembangunan Herba (HDD). Objektif utama kajian ini adalah bancian pengamal
perubatan Melayu di seluruh Semenanjung Malaysia berkaitan amalan dan
jenis tumbuhan ubatan yang digunakan dalam amalan mereka dan
menubuhkan koleksi ex situ tumbuhan ubatan. Projek ini juga membabitkan
kajian transliterasi manuskrip perubatan Melayu lama yang dikenali sebagai
Kitab Tib yang bermaksud himpunan ilmu perubatan (Pisol & Salahudin 2013).
Alatan Yang Digunakan Dalam Perubatan Tradisional Melayu
Peralatan yang digunakan dalam sediaan ubatan tradisional dan perawatan
merupakan elemen penting yang dipercayai mempengaruhi keberkesanan
sesuatu rawatan itu. Keberkesanan dalam konteks rawatan tradisional ini
membabitkan cerapan, kajian dan eksperimen, namun bukan seperti yang
ditakrifkan menurut sains moden yang sedia difahami oleh masyarakat kini
32
(Koharuddin 2003). Bahan asas seperti jenis batu, tanah dan besi bagi
menghasilkan alatan ini juga memainkan peranan berbeza dan memberikan
kesan terapeutik yang berlainan. Jenis penyakit yang ingin dirawat, jantina, usia
pesakit dan tahap keseriusan penyakit juga menentukan jenis alatan yang
sesuai digunakan untuk penyediaan ubatan dan merawat pesakit tersebut.
Kajian ini dijalankan khusus bagi pengumpulan maklumat dan kajian
alatan/artifak perubatan Melayu untuk mencapai 2 objektif iaitu:
1. Menghasilkan buku mewah “Alatan dalam Perubatan Tradisional Melayu”.
2. Mempamerkan koleksi alatan perubatan dan menubuhkan Muzium Alatan
Perubatan Melayu di Muzium Bioperubatan, IMR.
BAHAN DAN KAEDAH
Temu bual Pengamal Terpilih
Pemilihan pengamal yang memenuhi kriteria dibuat berdasarkan analisis ke
atas senarai pengamal daripada bancian awal yang dijalankan oleh FRIM.
Pengamal terpilih ditemubual bagi mendapatkan maklumat terperinci
berkaitan proses rawatan dan peralatan yang digunakan dalam amalan
mereka. Pemilihan pengamal dibuat berdasarkan kriteria berikut:
1. Menggunakan sebarang alatan penyediaan ubatan atau dalam rawatan.
2. Jenis dan cara rawatan serta peralatan yang digunakan samada unik atau
istimewa.
3. Rawatan dan alatan yang biasa tetapi dengan variasi jenis, saiz dan cara
penggunaan.
Aktiviti temu bual ini dijalankan dari September 2014 hingga Ogos 2015
ke atas seramai 65 orang pengamal yang dipilih dari seluruh Semenanjung
Malaysia. Pengamal ditemubual berpandukan borang soal selidik khusus sama
ada di rumah atau di premis perkhidmatan mereka. Demonstrasi berkaitan
juga divideokan. Temu bual ini merekod dan mendokumentasikan sejarah
amalan dan pengetahuan, termasuk maklumat sumber pengetahuan, jangka
masa mengamal, dan penerangan berkaitan amalan khusus mereka.
Perolehan Peralatan
Kajian ini juga mengumpul alatan dan artifak berkaitan sebagai koleksi dan
artifak khusus Alatan Perubatan Tradisional Melayu untuk dipamerkan di
Muzium Bioperubatan, IMR. Pengumpulan alatan dilakukan serentak atau
selepas mendapat maklumat melalui temu bual dilakukan ke atas pengamal.
33
Hasil kajian ini adalah terbitan sebuah buku mewah dengan memuatkan
gambar dan penerangan peralatan terpilih yang bersesuaian.
Jenis-jenis Amalan
Seramai 65 orang pengamal terpilih telah dicerap jenis amalan mereka melalui
temu bual yang dijalankan. Hasil temu bual mendapati amalan mereka boleh
dibahagi kepada kumpulan seperti dinyatakan di Jadual 1.
Jadual 1. Jenis amalan perubatan tradisional Melayu pengamal
Jenis amalan
Bilangan pengamal
Perbidanan
20
Urut patah dan biasa
14
Rawatan kerohanian
13
Penghasilan produk herba
10
Bomoh patah
7
Bekam
5
Bomoh umum
4
Resdung
2
Jumlah amalan mengatasi jumlah pengamal disebabkan ada pengamal
yang menawarkan lebih daripada satu jenis perkhidmatan.
Peralatan
Koleksi alatan yang diperolehi boleh dikategorikan mengikut jenis kegunaan
atau amalan berkaitan. Senarai jenis alatan adalah seperti di Jadual 2.
Jadual 2. Bilangan jenis alatan yang dikenalpasti
Jenis alatan
Penyediaan herba
Rawatan kerohanian
Perbidanan
Rawatan umum
Urut
Bomoh patah
Bilangan jenis
27
15
14
14
6
2
Peralatan yang berkaitan kebanyakkannya dibeli dari pengamal
(sekiranya dijual) atau diberi secara percuma oleh pengamal. Alatan yang tidak
34
berjaya diperolehi daripada pengamal akan dibeli di tempat lain, sama ada atas
cadangan daripada pengamal atau penyelidik mencari sendiri dengan bertanya
kepada orang yang berpengetahuan. Selain daripada itu, alatan yang tidak
boleh diperolehi dari mana-mana premis, alatan tersebut direplikasikan melalui
perkhidmatan pembuat replika profesional. Foto yang menunjukkan dimensi
dan saiz alatan dijadikan rujukan bagi menghasilkan replika.
Buku mewah yang diterbitkan memuatkan isi kandungan berdasarkan
maklumat daripada pengamal dan kajian keperpustakaan serta gambar-gambar
alatan berkaitan.
KESIMPULAN
Pelbagai maklumat yang diperolehi melalui kajian ini amat penting dipelihara
dalam bentuk pendokumentasian kerana ini merupakan khazanah warisan
bangsa yang amat bernilai. Selain daripada memantapkan identiti Melayu
dalam bidang perubatan tradisional, kajian seperti ini dapat mencetuskan
penemuan baru dalam bidang perubatan moden.
PENGHARGAAN
Penyelidik amat berbesar hati dan mengucapkan setinggi-tinggi penghargaan
atas kerjasama, sumbangan tenaga dan idea serta perkongsian maklumat oleh
pasukan penyelidik FRIM, Bahagian Perubatan Tradisional dan Komplementari
KKM. Tidak dilupai juga agensi lain yang bekerjasama dalam projek ini,
pembantu penyelidik IMR dan terutamanya para pengamal yang memberi
kerjasama yang baik dalam perkongsian ilmu mereka.
RUJUKAN
Editor, DBP. (2013). Kamus Dewan Edisi Keempat. Cetakan Keempat. Dewan
Bahasa dan Pustaka. Malaysia Dewan Bahasa dan Pustaka 2013.
Pisol, M. & Salahudin Suyorno, S. (2013). Etnofarmakologi Melayu-Islam:
Kearifan Tempatan dalam Manuskrip-manuskrip Melayu. Seminar
Serantau Kajian Manuskrip Melayu dan Kearifan Tempatan.
Koharuddin, M.B. (2003). Sains Perubatan Naturalistik Melayu: Satu Kajian
Awal. Jurnal Kemanusiaan FPPSM, UTM. Bil. 2/Disember 2003.
35
PENYELIDIKAN DAN PEMBANGUNAN SERTA PENGKOMERSIALAN
(R&D&C) PRODUK BERASASKAN TANAMAN UBATAN DAN BERAROMA
DI MARDI
WM Wan Zaki1, Z Mohd Hafizudin2, MA Mohd Shukri3, A Norzihan4, J Indu
Bala5, S Roowi6 & H Norma1
1
Pusat Penyelidikan Hortikultur, 2Pusat Penyelidikan Sains Ekonomi, 3Pusat
Penyelidikan Gen Bank dan Bijibenih, 4Pusat Penyelidikan Bioteknologi,
Penyelidikan 5Pejabat Ketua Pengarah MARDI, 6Pusat Sains Makanan, Ibu
Pejabat MARDI, Persiaran MARDI-UPM, 43400 Serdang, Selangor
Tel: 03-8953 6263
ABSTRAK
Kemajuan pesat industri herba negara perlu dimanfaatkan oleh semua pihak
terlibat agar komoditi ini dapat dijadikan sumber baru ekonomi negara. Usaha
yang tidak mengenal jemu semua pihak perlu digalakkan untuk mengambil
peluang memasukkan sains ke dalam herba tempatan agar kualiti bahan
mentah dapat dikekalkan pada aras yang tinggi dan keberkesanan serta
keselamatan produk herba dapat dibuktikan secara saintifik. Sehubungan itu,
dalam menunaikan tanggungjawab yang dimandatkan, MARDI sebagai sebuah
agensi di bawah Kementerian Pertanian dan Industri Asas Tani telah dan akan
melaksanakan tanggungjawab tersebut melalui program penyelidikan dan
pembangunan serta pengkomersilan (R&D&C). Ini meliputi bidang
pembaikbakaan, agronomi, pengurusan serangga dan penyakit, pengendalian
lepas tuai, pemprosesan primer dan sekunder serta pembangunan produk
makanan dan bukan makanan berasaskan herba bagi menambah nilai produk
herba tempatan. Pembaikbakaan bertujuan untuk mengumpul dan memilih
germplasma herba yang berkualiti agar tidak pupus. Pengeluaran bahan
tanaman berkualiti yang dipilih melalui proses pemilihan yang intensif juga
termasuk dalam aktiviti pembaikbakaan. Agronomi pula merangkumi semua
aspek pengurusan tanaman termasuk penyelidikan jarak tanaman, keperluan
baja, serta lain-lain keperluan asas hidup tumbuhan seperti air dan cahaya.
Pengurusan serangga dan penyakit yang berkesan pula merupakan faktor
kritikal untuk pertumbuhan pokok yang baik yang boleh merangsang hasil
biomassa dan kandungan fitokimia yang tinggi. Bahan mentah yang banyak dari
peringkat ladang perlu dikendalikan dengan cekap di peringkat lepas tuai bagi
memastikan kualiti yang diperoleh semasa dikutip dapat dikekalkan sehingga
penyimpanan. Pemprosesan primer dan sekunder yang cekap pula dapat
memastikan hasil dan kualiti bahan mentah dapat dikekalkan sehinggalah
proses pengumpulan bahan mentah untuk pembuatan produk. Apabila hasil
dan kualiti bahan mentah yang dipulihara sejak dari pengutipan dapat
digunakan sehingga proses pembangunan produk dapat memastikan produk
36
yang dikeluarkan sentiasa mempunyai aras kualiti yang tinggi. Namun begitu,
program penyelidikan yang dirangka ini hanya boleh berjaya apabila diberi
sokongan teguh oleh agensi utama sama ada dengan mewujudkan dan
memperkemaskan polisi, akta dan peraturan yang menyokong perkembangan
industri herba berserta peruntukan bajet yang mencukupi.
Kata kunci: Penyelidikan dan pembangunan, pengkomersialan, tanaman
ubatan dan beraroma, MARDI
PENGENALAN
Industri herba telah menjadi satu fenomena sejak dua dekad lalu berikutan
peningkatan kesedaran dalam konsep pengurusan kesihatan, penambahbaikan
kualiti produk serta bukti-bukti saintifik terhadap keberkesanan dan
keselamatan produk. Lantaran itu, industri herba tempatan perlu dimajukan
untuk mengambil peluang yang terbuka itu. Warisan biologi negara yang kaya
dengan spesies (t)anaman (u)batan dan (b)er(a)roma (TUbA = medicinal and
aromatic plants, MAP = herba) telah membolehkan Malaysia menjadi negara
penyumbang yang siginifikan kepada industri herba.
Di Malaysia, nilai dagangan tahunan bagi herba untuk tahun 2009–
2013 telah berkembang pada kadar 10% bagi import dan 21% bagi eksport.
Nilai import dan eksport tertinggi telah direkodkan pada tahun 2013 dengan
masing-masing bernilai USD470.51 dan USD110.83 (Comtrade, 2015). Fakta ini
menunjukkan pertumbuhan pesat industri herba di mana nilai eksport tumbuh
pada kadar lebih tinggi daripada import walaupun dalam keadaaan defisit
dagangan. Jumlah nilai import yang melebihi empat kali ganda berbanding nilai
eksport dengan jelas menunjukkan bahawa nilai dan kuantiti bahan mentah
yang digunakan dalam industri herba tempatan adalah tinggi dan kebanyakan
diperoleh dari sumber luaran. Sumber import yang utama ialah China,
Singapura, dan India yang majoritinya diimport oleh pedagang ubatan
tradisional Cina, Melayu dan India.
Penggunaan TUbA dalam perubatan tradisional Melayu telah mula
dikesan semasa pendudukan British yang mana manuskrip terawal mengenai
penggunaan ethnobotani TUbA Malaysia telah diterbitkan oleh Ridley (1907),
Gimlette (1913), Gimlette dan Burkill (1930), Burkill (1935), Gimlette dan
Thomson (1939) dan Gimlette (1971). Kesemua manuskrip tersebut menjadi
rujukan yang bererti terhadap penggunaan TUbA dalam perubatan tradisional
Melayu. Keadaan ini telah menjadi pendorong kepada penerusan aktiviti
penyelidikan saintifik yang lebih giat untuk menghasilkan produk-produk
tempatan yang berkesan dan selamat.
37
MARDI sebagai satu agensi di bawah Kementerian Pertanian dan
Industri Asas Tani telah dipertanggungjawabkan untuk melakukan penyelidikan
dan pembangunan serta pengkomersialan (R&D&C) untuk meningkatkan nilai
sepanjang rantaian pengeluaran TUbA. Usaha MARDI tidak akan berjaya tanpa
sokongan daripada jabatan dan agensi lain yang turut terlibat dalam industri
ini.
PERANAN MARDI DALAM R&D&C TUBA
Dalam usaha memartabatkan TUbA tempatan, MARDI telah melaksanakan
aktiviti R&D sepanjang rantaian nilai dari ladang ke meja. Bidang R&D&C yang
dilaksanakan termasuklah pembaikbakaan; agronomi; pengurusan perosak dan
penyakit; pengendalian lepas tuai; pemprosesan primer dan sekunder serta
pembangunan produk. Aktiviti tersebut dilaksanakan di beberapa pusat
tanggungjawab yang telah dikenalpasti mempunyai kepakaran serta peralatan
yang sesuai.
Pembaikbakaan
Bidang ini meliputi pengumpulan dan pemilihan germplasma TUbA untuk
mengekalkan ciri-ciri induk yang berkualiti serta bagi tujuan pemuliharaan agar
germplasma yang terpilih itu dapat meneruskan hidup dan tidak pupus. Usaha
pemuliharaan TUbA telah dijalankan sejak tahun 2000 di beberapa stesen
MARDI di Serdang, Jerangau, dan Bachok. MARDI Jerangau telah menjadi Pusat
Kecemerlangan (Centre of Excellence = COE) untuk pemuliharaan TUbA
manakala MARDI Bachok menjadi Pusat Kecemerlangan penyelidikan TUbA. Di
MARDI Jerangau, Pusat Gen Bank dan Bijibenih memelihara 131 spesies
dengan 138 aksesi sementara di MARDI Bachok pula mempunyai koleksi 18
spesies herba dan uniknya di sini ada koleksi 46 aksesi mas cotek yang
dipercayai terbanyak di negara ini.
Aktiviti pembaikbakaan sektor tumbuhan menggunakan platform
pengenotipan yang berskala tinggi juga terdapat di Pusat Penemuan dan
Validasi Molekul (Centre for Molecular Discovery and Validation = CMDV)
MARDI. Platform ini dapat menyokong pembangunan penanda molekul
kaedah Single Nucleotide Polymorphism (SNP) dan Simple Sequence Repeat
(SSR) melalui platform Sequenom, Illumina dan ABI 3730xl. Teknologi ini
diharap dapat menerajui dan meningkatkan bidang penyelidikan di Malaysia
supaya penghasilan baka yang baru dan berkualiti tinggi boleh dihasilkan dalam
jangkamasa yang lebih singkat. CMDV menawarkan perkhidmatan
pengekstrakan DNA berskala besar, pengecapjarian DNA, pembaikbakaan
molekul (Molecular Assisted Selection = MAS & Molecular Assisted Breeeding =
38
MAB), penemuan dan pengesahan penanda molekul, pengenotipan dan
latihan.
Agronomi
Bidang ini merangkumi aktiviti yang bermula daripada memperbanyakkan
bahan tanaman TUbA terpilih untuk memastikan kejayaan penanaman di
ladang samada berskala kecil, sederhana atau pun besar. TUbA yang dikenal
pasti berpotensi daripada sumber liar perlu didomestikasi terlebih dahulu agar
berjaya tumbuh dengan baik di luar dari persekitaran asal. Apabila proses
domestikasi ini berjaya, maka usaha menanam dalam skala lebih luas boleh
dilaksanakan. Bidang agronomi memfokuskan kepada aktiviti untuk
menentukan keperluan optimum jarak tanaman, keperluan baja, air serta
keperluan lain tanaman untuk memberikan pertumbuhan maksimum kepada
tanaman agar boleh memperoleh hasil biomassa serta kandungan fitokimia
yang tinggi. Pendapatan penanam dapat ditingkatkan apabila kombinasi hasil
biomassa serta kandungan fitokima berada pada tahap yang tinggi manakala
pemproses pula mendapat manfaat untuk mengeluarkan produk daripada
bahan mentah yang berkualiti tinggi.
Pengurusan Serangga dan Penyakit
Serangan serangga dan penyakit yang serius boleh mendatangkan kesan yang
sangat negatif kepada pertumbuhan tanaman yang seterusnya berupaya
menjejaskan hasil biomassa serta kandungan fitokimia dalam bahan mentah
TUbA. Penyelidikan terbaru di MARDI terhadap tanaman dukung anak telah
mengenalpasti penyakit utama bagi tanaman ini. Kaedah menentukan aras
infestasi dan keterukan penyakit telah dibangunkan. Penggunaan biopestisid
juga telah dikenal pasti berpotensi untuk mengawal perebakan insiden
penyakit pada tanaman ini. Penyelidikan terhadap perosak tanaman tongkat ali
pula telah mengenalpasti ulat harimau sebagai perosak penting yang berupaya
menjejaskan pertumbuhan pokok tongkat ali.
Pengendalian Lepas tuai
Amalan sebelum-kutip dan selepas-kutip yang kurang sempurna terhadap
TUbA boleh menjejaskan kualiti bahan mentah serta menurunkan sebatian
bahan aktif yang seterusnya menjejaskan kualiti produk siap. Faktor sebelumdan selepas-kutip yang diselidik termasuklah spesies yang sesuai, bahagian
pokok yang sesuai, masa yang paling sesuai untuk mengutip, parameter
optimum penanaman (tanah, cahaya, air, suhu dan nutrien), pengeringan dan
penyimpanan optimum, kontaminasi mikrob dan kimia, strategi pengekstrakan
optimum dan penyimpanan jangka panjang untuk produk siap. Isu kualiti dalam
39
produk siap boleh berlaku disebabkan spesies yang tidak sesuai, kutipan pada
tahap tumbesaran yang tidak optimum, kontaminasi oleh agen mikrob dan
kimia serta pengekstrakan yang tidak optimum.
Pemprosesan Primer dan Sekunder
Selepas dikutip, bahan mentah perlu diproses untuk memastikan kualiti dapat
dikekalkan. Apabila pemprosesan melibatkan TUbA jenis minyak pati, bahan
yang dikutip perlu diproses dengan mengerat atau mericik daun/batang
menjadi saiz yang lebih kecil untuk diproses ke peringkat seterusnya. Dalam
pemprosesan sekunder, bahan mentah tanaman minyak pati akan diproses di
dalam tempat pemprosesan yang sesuai seperti penyulingan hidro agar hasil
minyak pati dapat diperoleh pada kadar pengekstrakan yang tinggi.
Pembangunan Produk
Produk TUbA boleh dibahagikan kepada dua kategori iaitu makanan dan bukan
makanan. Produk makanan boleh berupa makanan tambahan atau
nutraseutikal untuk tujuan penjagaan kesihatan. Kategori kedua ialah produk
bukan makanan seperti aromaterapi, toiletries, biopestisida berasaskan TUbA.
Minyak pati misalnya, boleh diniagakan dalam bentuk 100% tulen tanpa perlu
diproses lagi. Minyak pati juga boleh digunakan sebagai satu daripada ramuan
dalam produk aromaterapi dan boleh juga digunakan dalam pembuatan
produk penjagaan diri seperti sabun dan gel mandian berasaskan minyak
tersebut. Minyak serai, serai wangi dan jeremin telah dibangunkan sebagai
biopestisida bagi menangani masalah perosak tanaman kekupu belakang intan
(Plutella xylostella) dan ulat ratus (Spodoptera spp) yang banyak menyerang
sayuran seperti kobis dan sawi. Produk biopestisida ini sedang diuji di peringkat
kajian lapangan. Beberapa minyak pati utama yang lain seperti nilam, gelam
dan gajus (Anacardium occidentale) juga telah digunakan untuk
membangunkan produk nilai tambah seperti sabun, syampu dan formulasi antikuman.
Pengkomersialan
Usaha telah dilakukan oleh MARDI untuk menggunakan teknologi moden bagi
menghasikan produk yang boleh dikomersialkan. Dalam RMKe-10, usaha ini
telah berjaya menghasilan beberapa produk yang berjaya dikomersialkan.
Contoh produk tersebut ialah NutrimaTM – Just Great (Roowi et al. 2013, Roowi
et al. 2012, Roowi & Crozier 2011, Roowi et al. 2010, Roowi et al. 2009 dan
Roowi 2008) dan ekstrak dukung anak (Lee et al. 2013). Produk-produk
40
tersebut telah berjaya memenangi beberapa anugerah dari dalam dan luar
negara.
Walaupun dalam banyak kes, khasiat ubatan yang diwar-warkan untuk
produk keluaran pengusaha tempatan kebanyakannya disandarkan kepada
testimoni sahaja dan tidak ada bukti saintifik, namun mutakhir ini usaha untuk
melakukan ujian keberksanan dan keselamatan produk secara saintifk telah
mula dilaksanakan.
Program inkobator-inkubati minyak pati di MARDI Kuala Linggi, Melaka
pula melatih enam usahawan tempatan dalam pengekstrakan dan
pembangunan produk berasaskan minyak pati (Jadual 1). Syarikat akan
mengikuti latihan di premis MARDI mengikut tempoh yang telah ditentukan.
Apabila telah berjaya menamatkan latihan masing-masing, syarikat akan
membuka kemudahan pemprosesan dan pembangunan produk sendiri.
Keenam-enam syarikat inkubati ini telah mempunyai produk dan berjaya
mengkomersialkan produk mengikut pelan syarikat masing-masing.
Kebanyakan syarikat memasarkan produk untuk pasaran tempatan manakala
satu syarikat telah berjaya memperoleh pasaran untuk minyak pati tulen lemon
myrtle ke Australia.
Jadual 1. Senarai syarikat yang mengikuti program inkubator-inkubati minyak
pati di MARDI.
Nama syarikat Inkubati
EssfaAgriculture Sdn Bhd
Tanaman
Gelam serai, serai wangi,
nilam
Janji Jitu Sdn Bhd
Serai wangi, gelenggang,
sireh, bunga raya
Serai wangi, gelam , kayu
manis
Excellent Wisdom
Qzen Plantation Sdn Bhd
Lemon myrtle
One Gifted Icon Sdn Bhd
Lemon myrtle
Pesona Herba Sdn Bhd
-
41
Produk
Gel mandian, losyen,
handwash berasaskan
minyak pati
Sabun, gel mandian, syampu
& penghalau serangga
Minyak pati, bar mandian,
gel mandian, losyen serai
wangi, penghalau serangga
Minyak pati lemon myrtle –
eksport ke Australia
Sabun, gel mandian, teh,
sachet wangian, serbuk
lemon myrtle
Gel mandian, sabun serai
wangi, gincu (serai wangi
dan mengkudu), lulur herba,
spa herba
KESIMPULAN
MARDI telah berusaha memenuhi mandat yang dipertanggungjawabkan
dengan melaksanakan aktiviti R&D&C merangkumi semua peringkat sepanjang
rantaian nilai pengeluaran TUbA daripada peringkat huluan hingga ke peringkat
hiliran termasuk melatih usahawan tempatan untuk menjadi usahawan yang
berjaya. Walau bagaimanapun, usaha ini tidak akan beroleh kejayaan tanpa
sokongan yang berterusan dari pihak berkuasa untuk merangka
polisi/akta/peraturan yang bermanfaat serta menyediakan peruntukan yang
mencukupi bagi pelaksanaan R&D&C terhadap TUbA, sesuai dengan hasrat
untuk membantu merealisasikan produk herba bernilai tinggi melalui EPP1
NKEA Pertanian.
RUJUKAN
Burkill, I. H. (1935). Dictionary of the Economic Products of the Malay
Peninsula. Crown Agents, London.
Comtrade.
(2015).
International
Trade
Statistics
http://comtrade.un.org/. Retrieved on Jan 2015.
Database,
Gimlette, J. D. (1913). Some Superstitious Beliefs Occurring in the Theory and
Practice of Malay Medicine. (Publisher unknown).
Gimlette, J. D. (1971). Malay Poisons and Charm Cures. Oxford University Press,
New York.
Gimlette, J. D. and Burkill, I. H. (1930). The Medical Book of Malayan Medicine.
Botanic Gardens, Singapore.
Gimlette, J. D. and Thomson, H.W. (1939). A Dictionary of Malay Medicine.
Oxford University Press, London.
Lee, S.H., Tang ,Y.Q., Jaganath, I.B., Sekaran, S.D. et al (2013). Effects of cocktail
of four local Malaysian medicinal plants (Phyllanthus spp.) against
dengue virus 2. BMC Complement Altern. Med. 26;13:192. doi:
10.1186/1472─6882─13─192.
Ridley, H. N. (1907). Materials for a Flora of the Malayan Peninsula. Methodist
Publishing House, Singapore.
Roowi, S. and Crozier, A. (2011). Flavonoids in tropical citrus species. Journal of
Agricultural and Food Chemistry 23;59(22):12217─25.
42
Roowi, S., William, M. and Crozier, A. (2012). Free phenolic acids in human
urine after drinking coffee rich with chlorogenic acids. J. Trop. Agric. And
Fd. Agric. (402):221-232.
Roowi, S., Hussin, Z., Othman, R., Muhammad, S. A., and Jusoh, A.Z. (2013)
Phenolic acid in selected tropical citrus. Asian Journal of Plant Biology
1:1─5.
Roowi, S (2008).Tropical Citrus Antioxidants and Catabolism of Phenolics in
Green Tea, Coffee, Cocoa and Orange Juice. Ph.D Thesis, University of
Glasgow, UK.
Roowi, S., Mullen, W., Edwards, C. A. and Crozier, A. (2009). Yoghurt impacts
on the excretion of phenolic acids derived from colonic breakdown of
orange juice flavanones in humans. Mol. Nutr. And Food Res. 53 (S1):68 –
75.
Roowi, S., Stalmach, A., Mullen, W., Lean, M. E.J., Edwards, C. A. and Crozier, A.
(2010).Green Tea Flavan-3-ols: Colonic Degradation and Urinary
Excretion of Catabolites. J. Trop. Agric. And Fd. Chem. 58 (2)─1296–1304.
43
KAJIAN RANTAIAN NILAI INDUSTRI HERBA TERPILIH DI SEMENANJUNG
MALAYSIA
AR Rohana, AB Ariff Fahmi, Z Nur Fazreen, A Nur Syazni, S Siti Zubaidah, R
Rosniza, M Marzalina, HF Lim, MA Mohd Shahidan, MA Nor Azah, O Zahari,
WY Wan Zahiri, K Rosdi & KY Pin
Tel: 03-62797559
Faks: 03-62736587 E-mel: [email protected]
ABSTRAK
Industri herba telah dikenal pasti sebagai salah satu pemacu kegiatan ekonomi
yang berpotensi memberi sumbangan penting kepada pertumbuhan ekonomi
Malaysia bagi mencapai status negara maju menjelang tahun 2020 di bawah
Program Transformasi Ekonomi (ETP). Sehingga kini, maklumat berkaitan
industri herba kurang diselaras dan dikoordinasi. Sehubungan itu, masih tiada
satu pangkalan data yang mengandungi maklumat komprehensif industri
herba. Bagi memenuhi aspirasi ini, kajian berkaitan rantaian nilai industri herba
adalah penting dan meliputi aspek permintaan dan penawaran. Rantaian nilai
industri herba merangkumi pembekal bahan tanaman, penanam, pengusaha,
pemborong dan peruncit. Kajian ini dijalankan bagi membangunkan pangkalan
data industri herba di peringkat huluan dan hiliran melalui bancian
komprehensif di seluruh Semenanjung Malaysia. Konsep rantaian nilai oleh
Micheal Porter menjadi asas kajian ini. Hasil bancian mendapati sebanyak
6,570 individu dan syarikat terlibat dalam rantaian nilai industri herba. Hasil
kajian ini dapat membantu dalam pembuatan keputusan bagi merencana hala
tuju industri herba berasaskan maklum balas langsung dari pihak industri
herba.
Kata kunci: rantaian nilai, industi herba, bancian
PENGENALAN
Perkataan ‘rantaian nilai’ mula digunakan oleh Micheal Porter di dalam
bukunya yang bertajuk Competitive Advantage: Creating and Sustaining
superior performance, 1985. Analisis rantaian nilai menggambarkan aktiviti
yang berkaitan dengan pencapaian sesebuah organisasi/industri sama ada
secara dalaman atau luaran. Ianya membolehkan kita menilai kekuatan dan
strategi organisasi/industri dalam penyampaian output atau perkhidmatan
kepada pelanggan di pasaran. Konsep rantaian nilai sebagai kaedah yang
44
menyokong pembuatan keputusan telah digunakan dalam strategi daya saing
yang dibangunkan oleh Porter pada awal tahun 1979.
Dengan mengambilkira keperluan serta kesukaran untuk mendapatkan
maklumat lengkap di setiap peringkat rantaian, pihak FRIM mengambil inisiatif
bagi menjalankan kajian rantaian nilai industri herba terpilih di Semenanjung
Malaysia dengan objektif-objektif seperti berikut:
1. Membangunkan pangkalan data industri herba di peringkat huluan dan
hiliran.
2. Mengenal pasti permintaan dan penawaran bahan mentah dan hasilan
produk herba.
3. Mengenal pasti jurang (gap) dalam landskap industri herba.
4. Mengenal pasti isu dan cabaran dalam industri herba.
BAHAN DAN KAEDAH
Projek ini melibatkan aktiviti-aktiviti berikut:
vi. Pengumpulan data sekunder dari pelbagai agensi
Setiap agensi yang berkaitan dengan industri herba dihubungi bagi
mendapatkan maklumat asas yang sedia ada di setiap jabatan. Maklumat
ini dikumpul dan disemak untuk tujuan verifikasi semasa sesi bancian.
Sumber data diperolehi daripada Jabatan Pertanian Semenanjung
Malaysia (2013) dan Kementerian Pertanian dan Industri Asas Tani
(2011).
vii. Bancian industri herba di Semenanjung Malaysia
Bancian dilakukan dengan melalui semua jalan yang ada di seluruh
Semenanjung Malaysia menggunakan peta, navigator dan GPS. Bancian
dilakukan secara bersemuka dan temu bual separa berstruktur
dilaksanakan berdasarkan borang soal selidik.
viii. Bengkel stakeholders consultation industri herba
Bengkel tersebut telah diadakan pada 24 Februari 2015 bertempat di
Batu Pahat, Johor telah mengumpulkan pelbagai pihak berkepentingan
di dalam industri herba. Input penting dari kesemua pihak dikumpulkan
bagi merangka strategi pembangunan industri herba Malaysia.
ix. Soal selidik/survei berkaitan sosio ekonomi industri herba
Berdasarkan hasil bancian, pengamal yang terlibat dalam aktiviti ini
dikenal pasti. Survei dijalankan mengikut kaedah “stratified random
sampling”. Temu bual separa berstruktur secara bersemuka telah
dilaksanakan berdasarkan borang soal selidik. Borang soal selidik ini
45
berkaitan 5 aktiviti utama dan 4 aktiviti sokongan berdasarkan Teori
Michael Porter seperti berikut:
Aktiviti utama:
1. Inbound logistics: aktiviti berkaitan penerimaan, penyimpanan dan,
pengagihan input kepada output meliputi pengurusan material,
pegudangan, kawalan inventori dan jadual pengangkutan.
2. Operasi: aktiviti berkaitan menukarkan input kepada produk akhir
seperti mesin, pembungkusan, pemprosesan, penyelenggaraan
peralatan dan pengujian.
3. Outbound logistics: Aktiviti berkaitan pengumpulan, penyimpanan dan
pengedaran produk kepada pembeli seperti penyimpanan produk akhir,
jadual pengangkutan dan penghantaran serta proses tempahan.
4. Pemasaran dan jualan: Aktiviti berkaitan penyediaan faktor yang
mendorong pembeli membeli produk seperti pengiklanan, promosi,
pemilihan saluran pemasaran dan harga.
5. Perkhidmatan: Aktiviti yang berkaitan penyediaan perkhidmatan bagi
meningkatkan atau mengekalkan nilai produk seperti waranti dan
demonstrasi penggunaan.
Aktiviti sokongan:
1. Perolehan: pembelian input untuk kegunaan keseluruhan rantaian nilai
seperti bahan mentah, mesin dan peralatan, tuntutan perbelanjaan dan
tatacara perolehan.
2. Pembangunan teknologi: Semua aktiviti berkaitan penambahbaikan
produk dan proses seperti teknologi dan prosedur know-how,
penyelidikan dan pembangunan produk, dan cara tempahan.
3. Pengurusan sumber manusia: tatacara pengambilan pekerja, bilangan
sumber manusia mengikut jenis, latihan dan kos sumber manusia.
4. Infrastruktur: merangkumi aktiviti pengurusan, perancangan,
perakaunan, perundangan, perhubungan kerajaan dan pengurusan
sistem maklumat iaitu overhead.
x. Pembangunan pangkalan data industri herba
Pangkalan data secara interaktif bagi menyimpan dan memaparkan
maklumat rantaian nilai industri herba dibangunkan menggunakan
platform berasaskan web. Selain fungsi kemasukan dan paparan data
bancian serta survei, pangkalan data tersebut turut memaparkan
pelbagai maklumat statistik berkaitan.
46
xi. Pembangunan laman sesawang kajian
Bagi tujuan promosi serta penyebaran maklumat kajian, pautan laman
sesawang projek melalui laman sesawang FRIM telah dibangunkan.
Kesemua informasi terkini berkaitan kajian dikemaskini dari masa ke
semasa di laman tersebut.
Ringkasan kaedah penyelidikan yang digunakan dalam kajian seperti berikut:
Sumber: Sekaran (2000), Wagner et al. (2012) & Rohana et al. (2015)
Jadual 1 menunjukkan bilangan responden bagi peringkat rantaian berbeza
iaitu pembekal bahan tanaman, penanam, pengusaha, pemborong dan
peruncit mengikut negeri di Semenanjung Malaysia (Rohana et al. 2015).
Kumpulan peruncit merupakan kumpulan responden teramai membentuk 78%
daripada keseluruhan jumlah responden dan majoriti tertumpu di bandarbandar besar.
Secara keseluruhannya, sejumlah 6,570 responden ditemu bual
sepanjang bancian dijalankan. Sebanyak 6,178 responden (94%) memainkan
peranan sebagai salah satu aktor rantaian nilai, manakala 392 responden (6%)
memainkan lebih daripada 1 peranan (Jadual 2). Selain responden yang
memberikan kerjasama, bilangan responden yang tidak bekerjasama dan
responden yang tiada di lokasi perniagaan sewaktu bancian dijalankan turut
direkod (Jadual 3). Peruncit, pemborong dan pengusaha merupakan kumpulan
47
sasar yang sukar untuk memberi maklumat sewaktu bancian, dengan masingmasing 14%, 10% dan 9% menolak untuk bekerjasama sewaktu ditemu bual
berbanding penanam dan pembekal bahan tanaman yang hanya 4% dan 3%
menolak sewaktu ditemu bual.
Jadual 1. Bilangan responden bancian mengikut kelas rantaian dan negeri di
Semenanjung Malaysia
Pembekal
bahan
Penanam Pengusaha Pemborong
Peruncit
tanaman
Perak
21
67
54
35
621
Selangor
25
83
119
100
1,166
Pahang
10
110
24
18
304
Kelantan
4
29
52
34
332
Johor
7
69
66
66
1,008
Kedah
5
19
74
52
403
Melaka
7
27
34
17
178
Negeri
28
21
16
294
Sembilan
Pulau
2
19
51
25
344
Pinang
Perlis
4
8
4
65
Terengganu
Kuala
Lumpur
Jumlah
8
7
9
20
192
1
-
22
48
583
90
462
534
435
5490
Jadual 2. Bilangan responden mengikut bilangan peranan/fungsi dalam
rantaian nilai industri herba
Bilangan
1 peranan/fungsi
6,178
2 peranan/fungsi
349
3 peranan/fungsi
37
4 peranan/fungsi
6
Jumlah
6,570
48
Jadual 3. Bilangan responden yang bekerjasama, tidak bekerjasama dan
individu tiada di lokasi perniagaan sewaktu bancian
Individu
Tidak
tiada/kedai
Bekerjasama
Jumlah
bekerjasama
ditutup semasa
bancian
Pembekal
86
3
1
90
bahan tanaman
Penanam
435
17
10
462
Pengusaha
451
50
33
534
Pemborong
378
44
13
435
4,497
766
227
5490
Peruncit
RUMUSAN
Maklumat yang diperolehi daripada kajian ini boleh digunakan bagi
menambahbaik penyelidikan dan pembangunan industri herba. Hasil kajian
yang diterbitkan membantu pihak yang berkepentingan (stakeholders)
membuat keputusan dan perancangan yang lebih baik dalam menentukan hala
tuju aktiviti penyelidikan, pembangunan dan komersialisasi. Selain itu,
maklumat ini juga dapat membantu pembentukan jalinan kerjasama di antara
pelbagai pihak berdasarkan pangkalan data yang telah dibangunkan.
RUJUKAN
Jabatan Pertanian Semenanjung Malaysia. (2013). Statistik Tanaman (Sub
Sektor Tanaman Makanan). Jabatan Pertanian Semenanjung Malaysia.
Putrajaya: Jabatan Pertanian Semenanjung Malaysia.
Kementerian Pertanian dan Industri Asas Tani. (2011). Dasar Agromakanan
Negara 2011─2020. Putrajaya: Bahagian Perancangan Strategik dan
Antarabangsa, MOA.
Porter, M.E. (1985). The Competitive Advantage: Creating and Sustaining
Superior Performance. Free Press, NY.
49
Rohana, A.R., Nur Fazreen, Z., Ariff Fahmi, A.B., Nur Syazni, A., Siti Zubaidah, S.,
Lim, H.F., et al. (2015). Directory of Herbal Cultivators in Peninsular
Malaysia. Kuala Lumpur: Forest Research Institute Malaysia.
Rohana, A.R., Nur Fazreen, Z., Ariff Fahmi, A.B., Nur Syazni, A., Siti Zubaidah,
S., Lim, H.F., et al. (2015). Pp. 211–219 in Value Chain Study for Holistic
Development of Herbal Industry. Proceeding 2015 3rd International
Conference on Rural Development & Entrepreneurship, Hebei Province,
China.
Sekaran, U. (2000). Research method for business─a skill building approach,
John Wiley & Sons, USA.
Wagner, C., Kawulich, B. & Garner, M. (2012). Doing social research: a global
context. McGraw-Hill Higher Education, UK.
50
MENGANGKAT TONGKAT ALI KE MATA DUNIA: PENGALAMAN
PENGKOMERSIALAN
MY Awang Ahmad
Kompleks Perhutanan, Fakulti Sains dan Sumber Alam, Universiti Malysia
Sabah, 88999, Kota Kinabalu, Sabah Malaysia
Tel: 088-320118
Faks: 088-320876
ABSTRAK
Kertas kerja ini akan membincangkan pengalaman penyelidik dalam
membangunkan produk berasaskan tongkat ali bermula dari peringkat
penanaman sehingga kepada produk akhir iaitu pada tahap pre-komersial dan
telah dikomersialkan di Malaysia. Masalah yang dihadapi dalam industri
berasaskan herba juga turut dibincangkan
Kata kunci: Tongkat ali, EurypotTM, pokok perhiasan, makanan tambahan
haiwan, pengkomersialan
PENGENALAN
Pasaran herba di Malaysia mengalami peningkatan yang mendadak sejak
kebelakangan ini. Keadaan ini disebabkan oleh kecenderungan masyarakat
untuk memilih produk herba sebagai alternatif kepada perubatan atau terapi
konvensional ataupun sebagai diet tambahan. Pada masa kini, penggunaan
herba sebagai bahan asas dalam perubatan tradisional dan juga moden
semakin meluas. Namun begitu, penggunaannya adalah lebih tertumpu kepada
perubatan tradisional dan dijual dalam pelbagai bentuk produk seperti bahan
kering, kapsul, tablet ataupun tonik minuman. Kegunaannya juga telah
dipelbagaikan sebagai bahan campuran dalam persediaan minuman dalam tin.
Berdasarkan kepada laporan Wiart et al. (2002), nilai pasaran herba telah
mencapai kepada RM 1 billion pada tahun 1998 dan dijangka akan terus
meningkat dari tahun ke tahun.
Tongkat ali ataupun nama santifiknya Eurycoma longifolia terkenal
diseluruh dunia sebagai salah satu bahan penambah tenaga bagi golongan
lelaki. Pandangan ini telah menyebabkan Tongkat Ali hanya digunakan ataupun
dibangunkan sebagai produk untuk kesihatan lelaki sahaja. Keberkesanan
tongkat ali memang tidak dinafikan, malah kajian klinikal juga telah
membuktikan akan keberkesanannya. Namun jika menumpukan kepada satu
segmen pasaran sahaja akan akan menghadkan kepelbagaian guna produk
51
tongkat ali. Keadaan ini boleh mengurangkan keuntungan kepada industri
herba di Malaysia, sekiranya ingin menembusi pasaran luar negara. Kertas kerja
ini akan membincangkan kegunaan lain tongkat ali selain dari kegunaannya
sebagai afrodisiak, dari pengalaman penulis serta kajian literatur.
KEGUNAAN TONGKAT ALI
Bhat dan Karim (2010) melaporkan tongkat ali mempunyai pelbagai jenis
kegunaan dari bahagian akar hingga ke daun. Banyak kajian berkaitan sifat
pharmakologi tongkat ali telah dilakukan. Antaranya akar, batang, dan kulit
mempunyai aktiviti anti plasmodial (anti malaria), manakala
daun
mengandungi aktiviti antitumor dan antimikrobial. Sebatian kimia juga telah
berjaya diipencilkan dari berbagai bahagian pokok tongkat ali. Namun demikian
bagi mendapatkan sumber mentah tongkat ali yang berkualiti, penghasilan
sumber yang boleh dipercayai dan mapan harus dititikberatkan. Sehubungan
dengan itu, penulis telah berjaya membangunkan satu teknologi penanaman
tongkat ali secara mapan iaitu EurypotTM.
Eurypot™
Di antara masalah penanam tongkat ali di Malaysia adalah keupayaan untuk
mendapatkan struktur akar yang lurus ke bawah, tumbesaran yang tidak
sekata, serangan dari ulat harimau pada daun dan juga menyebabkan
penghasilan buah yang kurang. Berdasarkan masalah-masalah tersebut,
Eurypot™ dibangunkan khas untuk penanaman tongkat ali. Eurypot™ ialah
sejenis pasu yang dibangunkan melalui dana penyelidikan dari Malaysian
Timber Industry Board (MTIB) bagi projek ladang hutan. Eurypot™ telah
didaftarkan sebagai tanda dagangan dan juga lukisan industri.
Melalui penggunaan Eurypot™, hasil yang diperolehi bagi bahagian akar
adalah lebih baik, iaitu lebih lurus, kerana Eurypot™ juga bersifat sebagai “roottrainer”. Manakala dengan menggunakan konsep tertutup, serangan dari
serangga dan penyakit dapat dikurangkan. Ini seterusnya dapat meningkatkan
produktiviti penanam tongkat ali terutama dalam penghasilan daun dan buah.
Kajian oleh Jiwajinda et al. (2001) telah mendapati bahawa daun
tongkat ali mempunyai aktiviti anti tumor dan anti parasitik. Melalui kajian ini
potensi daun tongkat ali dapat dibangunkan untuk penghasilan produk baharu.
Ini dapat meningkatkan nilai tongkat ali dan seterusnya menarik minat petani
untuk mengusahakan tanaman tongkat ali.
52
Tongkat Ali Sebagai Pokok Hiasan (Landskap)
Tanaman hiasan merupakan salah satu lagi cara untuk meningkatkan nilai
pokok tongkat ali. Pada masa ini, pokok tongkat ali belum dianggap sebagai
pokok hiasan dan belum disenaraikan dalam senarai tumbuhan hiasan di
bawah Jabatan Landskap Negara. Melalui kajian yang dijalankan oleh Program
Taman Alam dan Rekreasi di Universiti Malaysia Sabah (UMS), mendapati
bahawa, tongkat ali mempunyai bentuk monopodial sama seperti pokok
eksotik Brazilian Tower Tree yang sering digunakan sebagai pokok hiasan di
perbandaran di Malaysia. Walau bagaimanapun, kos pembelian pokok eksotik
ini adalah agak tinggi iaitu dalam lingkungan RM100-300 sepokok.
Berdasarkan kepada temu bual yang pernah dilakukan, ramai peminat
seni taman sanggup membayar RM30-100 sepokok bagi pokok tongkat ali pada
ketinggian 1-3 kaki. Ini kerana pokok tongkat ali amat mudah untuk dijaga dan
menghasilkan daun yang lebat dan mengambil masa yang lama untuk luruh.
Pokok tongkat ali juga tidak memerlukan ruangan yang besar, iaitu
berdasarkan saiz kanopi maksimum 1 meter diameter (Gambar 1). Disamping
itu tongkat ali juga menghasilkan bunga dan buah yang menarik.
Gambar 1: Pokok tongkat ali
53
Tongkat Ali Sebagai Makanan Tambahan Haiwan
Industri penternakan di Malaysia merupakan pelaburan besar yang bernilai
berbillion ringgit bagi memenuhi keperluan makanan negara. Antibiotik
diperlukan untuk memastikan ayam ternakan dalam keadaan yang sihat dan
membesar bebas daripada jangkitan penyakit. Penggunaan antibiotik dalam
penternakan ayam pada peringkat ladang boleh mencecah sehingga RM0.30
seekor. Namun, permintaan terhadap ayam bebas antibiotik juga semakin
meningkat, kerana haiwan yang diberikan antibiotik boleh memudaratkan
kesihatan.
Situasi ini telah membuka ruang kepada penggunaan herba tempatan
sebagai bahan tambahan dalam diet haiwan penternakan. Melalui kajian yang
telah dijalankan pada peringkat makmal sehingga peringkat komersial, didapati
bahawa tongkat ali mempunyai potensi untuk dikembangkan sebagai bahan
tambahan makanan bagi penternakan ayam di Malaysia (Gambar 2). Kajian
selanjutnya yang sedang dijalankan juga menunjukkan tongkat ali boleh
digunakan dalam industri haiwan kesayangan.
Gambar 2: Ayamli™ Betina 3kg, 42 Hari
54
KESIMPULAN
Walaupun pokok tongkat ali dinobatkan sebagai raja herba di Malaysia, namun
usaha untuk mempromosi dan meninggikan nilai tongkat ali masih lagi rendah
jika dibandingkan dengan ginseng Korea. Pelaburan jutaan ringgit dalam
penyelidikan tongkat ali harus diterjemahkan dalam bentuk hasil bermanfaat
kepada pengguna. Sehubungan dengan itu, sudah tiba masanya untuk
menghasilkan pelbagai produk tongkat ali, daripada penghasilan bahan mentah
sehingga kepada produk ubatan dan kosmetik yang bernilai tinggi. Inovasi
produk berasaskan tongkat ali hasruslah disokong dengan pemberian dana
yang setimpal.
RUJUKAN
Jiwajinda, S., Santisopasri, V., Murakami A, Hirai, N., Ohigashi, H. (2001).
Quassinoids from Eurycoma longifolia as plant growth inhibitors.
Phytochemistry 2001;58:959–62.
Bhat, R. And Karim, A.A. (2010). Tongkat Ali (Eurycoma longifolia Jack): A
review on its ethnobotany and pharmacological importance.
Fitoterapia 81(7): 669─679.
Wiart, C. (2002). Medicinal Plants of South East Asia. Prentice Hall, Pearson
Malaysia. Sdn.Bhd., Selangor Malaysia.
55
ANALYSING MALAY MEDICAL MANUSCRIPT MSS2999: THE FINDINGS
AND THE HOPE FOR THE FUTURE
H Abdul Ghani
Herbwalk Consultancy, 66, Batu 8, Jalan Ayer Hangat,07000 Langkawi, Kedah
Darulaman
E-mail: [email protected]
INTRODUCTION
Traditional Malay medicine is not a new after thoughts but rather, it is at the
very least more than 6,000 years old. It stems out from the need to treat
diseases of the community. Like most other traditional medicine, it is based on
the understanding of what is normal and what comprise abnormality. It is this
that directed the practitioner towards concocting the appropriate remedies to
correct the abnormalities. The practice initially was based on instinctual
observations using simple remedies comprising of single materia medica, to
the eventual understanding of the varied forces in the materia medica which
could help in enhancing the effects of the remedies prescribed. This had
resulted in the cooking up of more complex remedies utilising varied materia
medica to render the treatment better directed. The old practitioners indeed
are people with great knowledge and great minds for without them they could
not possibly have craftily concocted remedies very well directed to the cure of
the diseases being addressed.
THE MANUSCRIPT
MSS 2999 is a small manuscript of 22 pages with 141 prescriptions for the
treatment of 42 different diseases in 12 different anatomical regions. It was
obtained from Pusat Manuskrip Melayu of the National Library of Malaysia.
Not much is known of the origin or the author of the manuscript. It is probably
notes of a student during his learning process with his ‘guru’. This is reflected
upon the lack of organisation of the prescriptions according to system of
disease.
56
THE DISEASES
Most of the diseases addressed were trivial and it can be assumed the most
common ailments of the period the manuscript was written. It covers diseases
of the eye, ear, nose and throat, dental problems, gastrointestinal tract,
respiratory tract, urinary tract, fevers and infective processes and diseases of
women. There was no mention of treatment of chronic diseases like diabetes,
hypertension, cardiac diseases nor of cancer. It may be assumed that either
these diseases were not common or the practitioner then has no knowledge of
these disorders. The approach to treatment at times are very similar to modern
medical therapy while at time the approach differs but is deemed more
appropriate than those of modern therapy.
THE MATERIA MEDICA
There are a total of 120 unique materia medica of vegetable origin in 53
families being used. Most of the plants used are those easily obtainable around
the house and within close vicinity to village. None of the mentioned plants
were from the rainforest. Out of the 120 different plant materials, 24 were
those easily obtainable from the Indian spice shops. Most of these were
imported either from India or the Middle East. There are 36 items which we
classify as non-vegetative. There are 10 different liquids used in various
prescriptions including water, oils, honey, and various milks. Three types of
charcoal were used and they are made out of coconut shell, teak wood and
bottle gourd shell. Of gums and resins, there were six in total.
CONCLUSION
For a nation with unwritten history of more than 6,000 years, one cannot deny
the advancement of their knowledge. With the discovery of the archaeological
site at Sungai Batu, Kedah, it further provides evidence of this long history and
puts the Malay nation a being far more matured than most other nations with
technological advancement in the field of iron smelting as being the
oldest.Traditional Malay medicine too may have evolved to an advanced stage
with knowledge being spread to wherever they used to trade via the sea route.
Exchange of knowledge is a continuous learning process of this nation as can
57
be noticed from the use of materia medica from China, India, the Middle East
and Europe.
From the analysis of the manuscript many things could be concluded
about the general health of the people during the time of writing of the
manuscript, the common diseases of the time, the various methods of
preparation of the medicine, the choice of material medica, the use of the
concoctions for single purpose and the knowledge of diseases and the
medicine to treat them.
Further analysing old traditional Malay medical manuscript can open
new frontiers of research in the field of Malay herbal medicine. It can provide
well directed research on the values of plants as medicine as understood by
the Malay communities of the past.
58
COMMERCIALIZED INNOVATIVE HERBAL PRODUCTS FROM UPM
M Suhaila1, 2 & MN Fatihanim1
1
Functional Food laboratory, Faculty of Food Science and Technology
2
UPM─MAKNA Cancer Research Laboratory, Institute of Bioscience, University
Putra Malaysia, 43400 Serdang, Selangor, Malaysia
ABSTRACT
The product invention is an additive which can be incorporated into edible oil
composition especially cooking oil to provide therein antioxidant effect and
reduce oil adsorption into the food prepared. The additive is composed of
biophenolics formulated from plant extracts of the Rutaceae family. The
antioxidant characteristics of Rutaceae leaf alcohol extract were evaluated in
refined, bleached and deodorized (RBD) palm olein at 180oC using an
accelerated oxidation and deep frying studies for up to 40 hours. The extract,
at the optimum concentration 0.1%, retarded oil deterioration significantly
(P<0.05) in deep frying studies, comparable to 0.02% BHT (Butylated Hydroxy
Toluene) in tests such as peroxide value, anisidine value, free fatty acid,
Oxidative Stability Index (OSI), polar and polymer compounds content. The
sensory evaluation of french fries fried in oil containing the extract showed
significant (P<0.05) improvement in crispiness, taste and overall quality,
especially after the 40th hour of frying compared to those similarly fried in the
control oils and the oils containing BHT. The polyphenols content of the extract
were 103.2 mg/g gallic acid equivalent. Rutaceae leaf extract exhibited
excellent heat stable natural antioxidant properties that retarded oil
degradation and improved the quality of the fried product.
Keywords: frying, Rutaceae leaf extract, palm olein, sensory evaluation,
antioxidant, accelerated oxidation study, deep frying
INTRODUCTION
Deep frying at high temperatures hastens oxidation, oil deterioration and
various chemical reactions affecting the fried product taste, consistency, shelflife and nutrients. The presence of antioxidants extends oil lifespan and helps
maintain the product quality. At present, synthetic antioxidants such as
butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tertiary
butylated hydroxyquinone (TBHQ) are commonly used, but they are volatile
and unstable at frying temperatures. Synthetic antioxidant and natural
59
antioxidants have been proven to be capable of retarding oxidation in frying
and in accelerated oxidation study. Phenolic compounds such as catechin and
catechin composites have been shown to significantly improve the stability of
peanut oil and palm superolein frying performances. The objective of this study
was to evaluate the antioxidative characteristics of Rutaceae leaf extract in
palm olein during accelerated oxidation and frying.
MATERIALS AND METHODS
The cleaned Rutaceae leaves were dried in a hot air oven at 45oC for 24 hr,
ground to a fine powder before extracting with 10 times its weight in ethanol
for 8 hr at 50oC. The solvent was evaporated using rotary evaporator.
Frying Experiment
Deep frying was carried out in a stainless steel electrical open fryer. The
treatments conducted simultaneously were, (1) Afdhal-x, palm olein containing
0.1% Rutaceae leaf extract (10 kg oil was introduced into separate fryers, and
heated up to 60oC before adding 0.1% extract, and stirred until completely
dissolved) (2) palm olein containing 0.02% BHT, and (3) palm olein without any
additive as control. Approximately 400 g oil samples were collected from each
fryer to represent sample for day 0 before frying. The remaining oil was heated
at 180  2oC and was allowed to equilibrate at this temperature for 30 min.
About 14 batches at 200g per batch of french fries were fried for 2.5 min per
day at 30 min intervals for 8 hours daily. The fryers were turned off at the end
of the frying experiment each day and the oil cooled to 60oC, before filtering
using separate filters to remove debris. Accurately weighed 400g of the frying
oil from each fryer were sampled into amber bottles at the end of each day. All
oil samples were flushed with slow bubbles of nitrogen from the bottom of the
bottles and stored in at -20oC for physical and chemical analysis.
Sensory Evaluation
Sensory evaluation was conducted on the same day using the 5th and 6th
batches of fried French fries. The fryers were topped off up to 10 kg with oil
containing antioxidants (0.02% BHT or 0.1% Rutaceae leaf extract) depending
on the oil loss. The whole procedure was repeated consecutively for 5 days.
The French fries colour, flavour, oiliness, crispiness, taste and overall quality
were evaluated using a 9- point hedonic scale (1= very poor; 9= very good). The
sensory was done by 20 trained panellists on day 1, 3 and 5 equivalent to 8, 24
and 40 hours of frying in the same oil.
60
Statistical Analysis
Each experiment and analysis, including leaves sampling and extraction, were
conducted in triplicate. The MINITAB 14 software was used to analyse data for
determining ANOVA, standard deviation and Duncan’s multiple range test for
significance at a 5% level.
RESULTS AND DISCUSSION
Peroxide Value and Anisidine Value,
In deep frying studies the peroxides present in the oil containing the extract
(Afdhal oil) were lower than in the BHT, indicating that in the presence of food,
the compounds in the extract significantly (P<0.05) reduced peroxides
formation since less of the peroxides have changed to the secondary products
as confirmed by the anisidine values, when compared to the oils containing
BHT and the untreated control (Figure 1).
The Peroxide values (Figure 1) which measures the primary oxidation products
in the oil, is expected to initially increase and then decrease after certain hours
of heating, due to the formation of secondary oxidation products such as
ketones, aldehydes, hydrocarbons, epoxides etc. which were then gauged by
the anisidine values. Peroxide levels were evaluated in the frying oil from their
ability to liberate iodine from potassium iodide. Peroxides formed in palm oil
were relatively low (below 3m eq oxygen/kg oil after 30-40 hours at 180°C)
compared to other more unsaturated cooking oils because it contains about
50% monounsaturated fatty acids and about 45% saturated fatty acids with
very low levels of polyunsaturated fatty acids. Palm oil is also very rich in
tocotrienols, which act as endogenous antioxidants, making the oil fairly stable
to high temperatures use.
Sensory Evaluation
Sensory evaluation of French fries fried on the 40th hours in the various oils
showed significantly better overall quality, for samples fried in oils containing
Rutaceae leaf extract (Afdhal-x), unlike the control and BHT samples. However,
no significant difference (P<0.05) were found in scores for oiliness, crispiness
and taste observed between samples throughout the frying experiment (Figure
1). Evaluation on the colour of French fries shows that Rutaceae and control
samples were not significantly different (P<0.05) from one another during the
40 hours of frying. Although the colour of oil and French fries were darker in
the presence Rutaceae extract, and increases significantly during frying, the
colour sensory scores of French fries were the reverse. French fries fried in oil
containing Rutaceae extract were preferred as the number of frying hours
61
increased to the 40th hour (Figure 1). Samples fried in the control oil showed a
slight increase in the colour intensity with increasing frying time, as expected.
BHT caused slight decrease in scores as the frying hours increased. Flavour
scores for all samples were not affected by increasing frying time, but were
slightly different from one another by the 40th hour of frying. The mean scores
were not significantly different (P<0.05) because of the high standard
deviations in the scores given by the sensory panellists. All samples fried in oil
containing Rutaceae extract were preferred even at the 40th hour of frying
(average score of 7.00); unlike those fried in oil containing BHT which scored
lower than 5. The leaf extract improved the fry’s characteristics especially for
flavour scores after using the oil for 40 hours of frying.
62
The Oil Content of the Sample (%)
Oil content (%)
5
4
3.54
Nugget
3.10
3
2.85
2.61
2.52
2
2.51
2.30
1.74
1.38
1.35
1.22
1.13
1
1.26 1.12
0.96
0.38
French
fries
Keropok
Lekor
Doughnut
0
control
c.grandis
c.aurantifolia
c.microcarpa
Sample
Figure 1. Physicochemical characteristics of cooking oil and fried food products
with the use of the Afdhal cooking oil additive.
63
CONCLUSION
This study shows that Rutaceae leaf alcoholic extracts are effective at retarding
oil oxidation. The leaf extract maintained the sensory qualities of French fries,
better than BHT, in reused oil, showing that Rutaceae leaf is a good antioxidant
for oils, besides having other beneficial health properties.
ACKNOWLEDGEMENTS
The authors would like to thank Ministry of Education for financially supporting
the commercialisation of this work
REFERENCES
Mohamed, S. and Nor, F.M. (2008) Additive for edible oil composition. US
Patent App. 12/745,635.
64
DEVELOPMENT OF HERBAL PRODUCT FOR CANCER THERAPY
AH Loiy Elsir1, H Suzana1, AM Aman Shah2 & AM Amin Malik Shah1
1
EMAN Laboratory, Pharmacological Dept., School of Pharmaceutical Sciences,
USM, 11800 Minden, Penang
2
Advanced Medical and Dental Institute (IPPT), Universiti Sains Malaysia,
Penang, Malaysia
Tel: 04-653 4582
ABSTRACT
Herbal medicine, also called botanical medicine or phytomedicine is basically
the application of the plant material such as root, leaves, bark, flowers, seeds
and berries used in the maintenance of health, prevention, diagnosis and
improvement or treatment of the illness. Herbal medicine has been in
existence since ancient time and the oldest treatment system in the world. The
fact that plants had within them the naturally occurring chemicals and
compounds that effective in healing various illnesses is a key for herbal
medicine practice worldwide. The value of herbal medicine in preventing and
treatment of illness has increased due to improvement in analysis and quality
control as well as advances in scientific research. In this paper, we summarise
the development of the herbal product for cancer therapy.
Keywords: herbal medicine, cancer, natural product, chemical compounds,
treatment
INTRODUCTION
Cancer is a major public health burden in both developed and developing
countries. The International Agency for Research on Cancer (IARC), the
specialized cancer agency of the World Health Organization, reported that
about 14.9 million cancer cases were estimated around the world in 2013.
From these amount, 7.7 million cases were in men and 6.9 million in women
and further this number is expected to increase to 24 million by 2035 (Bray et
al. 2013). Treating cancer has become a major challenge as there is no single
effective treatment that works for all types of cancer. Most of conventional
chemotherapy regimens which employ different combinations of cytotoxic
drugs are often associated with serious side effects and chemo-resistance.
Conventional therapy has also become less favourable in the mindset of
sufferers, as a result many patients resort to seeking alternative treatments
(Chorawala et al. 2012).
65
The resistance of metastatic cancerous cells to chemotherapy and its
adverse effects has become a serious challenge in cancer research. Despite the
intensive progress in chemotherapeutics in the last decades, the need to
discover and to develop new, alternative, or adjuvant therapeutic agents
remains.
NATURAL PRODUCTS AND CANCER TREATMENT
Natural Products, especially plants, have been used for the treatment of
various diseases since antiquity to date. Secondary metabolites from plants
have such as flavonoids, alkaloids, terpenoids and glycosides have been prized
for their protective effect against different pathogens (e.g bacteria, fungi or
insects), pain-relieving and healing activity or growth regulatory molecules (e.g.
hormone-like substances). Also, Phytochemicals have a fundamental role in
prevention and treatment of cancer either by modulating the activity of
specific hormones and enzymes responsible for carcinogenesis or by direct
cytotoxicity against cancer cells (Kintzios & Barberaki 2004). Natural products
have been forefront in cancer treatment; recently pharmaceutical companies
have explored more than 25,000 plants for anti-cancer drugs. About 60% of the
anti-cancer drug over the globe have been discovered from plant origin
between 1983 and 1994 (Sakarkar & Deshmukh 2011). Phytoconstituents
derived from plants are paclitaxel from Taxus brevifolia Nutt, vinblastine
and vincristine from Catharanthus roseus, etoposide and teniposide which
are epimers of podophyllotoxin that isolated from the roots of different
podophyllum species, also camptothecin which is extracted from the Chinese
tree Camptotheca acuminata (Newman et al. 2000). The significance of
phytoconstituents in treatment of cancer is very obvious, for instance the
discovery of paclitaxel (Taxol®) evident the success of natural products in
combating cancers (Oberlies & Kroll 2004, Cragg & Newman 2005).
It’s well known that the growth of solid tumors depends mainly on
angiogenesis. Many natural compounds proved to have antiangiogenic effects
such as asisoliquiritin and combrestastin were isolated from Glycyrrhiza glabra
and Combretum caffhum respectively were found to inhibit MMPs and tubules
formation, in turn interrupt angiogenesis (Pettit et al. 1989, Kang et al. 2010).
Other natural compounds showed potent antiangiogenic activity including
resveratrol from grapes (Cao et al. 2005) quercetin (Tan et al. 2003),
rosmarinic acid (Huang & Zheng 2006), genistein (Su et al. 2005) and curcumin
(Gururaj et al. 2002) with various mechanism of actin. Some compounds
modulates VEGF signalling pathway either by decreasing the production of
VEGF or interfering with binding of VEGF to its receptors such as Vincristine,
Ginsenosides, Taxol and Triptolide extracted from V. rosea, P. ginseng, T.
brevifolia and T. wilfordii respectively (Attardo & Sartori 2002). Several flavones
66
and flavonols isolated from different plants reported to inhibit endothelial cell
proliferation and migration the key steps in angiogenesis (Fotsis et al. 1997).
Plant as sources of antiangiogenic agents could be of great value, as they can
be produced commercially in large quantities (Al-Suede et al). Although, there
is no plant based antiangiogenic compounds available commercially at present.
It is known that reactive oxygen species such as superoxide (O2˙-),
hydroxyl (OH˙-), and peroxyl (˙OOH, ROO˙) radicals are produced under
oxidative stress and 5% of inhaled oxygen is converted to reactive oxygen
species. Reactive oxygen species have been implicated in pathological and
degenerative ailment such as cancer, aging, coronary heart disease,
Alzheimer’s disease, diabetes, and inflammation(Pham-Huy et al. 2008).
Antioxidants are substances that neutralize the oxidative effect of free radicals
via different ways, thus it can protect the body from some diseases. Vitamins
(C, E carotenoids, etc.), flavonoids (flavones, isoflavone, flavonones,
anthocyanins and catechins), polyphenols (ellagic acid, gallic acid and tannins)
from plants demonstrate incredible antioxidant activity (Gupta & Sharma
2006). Parthenolide, phytochemical extracted from the antioxidant herb,
Feverfew showed anti-inflammatory potential through inhibition of IkB kinase,
a cytokine-mediated stimulation of genes involved in inflammation (Pal et al.
2014).
USING HERBS AS ANTI CANCER AGENTS
An herb is a plant or plant part used for its scent, flavor, or therapeutic
properties. Herbal medicines are dietary supplement used to treat various
diseases. Recently, traditional medicine such as Chinese medicine, Kampo
medicine and Ayurveda are used in Asian countries and most of them are
accepted as popular complementary and alternative medicine therapies for
cancer worldwide (Elujoba et al. 2005). In some countries, herbal medicines are
in some forms of drugs listed in Chinese and Japanese pharmacopoeias now
used in daily cancer clinical practice. It is common approach in herbal medicine
studies nowadays to find active herb in a composite formula for certain disease
and further explore active ingredients in the herb, thereafter composite
formula can be standardized for biological activity and chemical composition.
FDA classify the drug products from herbal medicine as dietary supplements
and botanical drug products, with regard to the experiences and problems in
herbal medicines for its use as a single and as combination (Feng et al. 2011).
Botanical products from herbal medicines are extracts/fractions containing
several active components which can be determined precisely using chemical
analysis technologies such as gas chromatography (GC), high-performance
67
liquid chromatography (HPLC) and mass spectrophotometer (MS)(Choi et al.
2009)
CONCLUSION
Herbal product is believed to offer prophylaxis (chemoprevention) or a
relatively safer treatment option with a minimum side effect. Development of
herbal product is undeniably one of the options for maintenance of health,
prevention, diagnosis and improvement or treatment of the illness, especially
for the cancer therapy.
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Strategy Products. Bollettino chimico farmaceutico 142(2): 54─65.
Bray, F., Ren, J. S., Masuyer, E. & Ferlay, J. (2013). Global Estimates of Cancer
Prevalence for 27 Sites In The Adult Population In 2008. International Journal
of Cancer 132(5): 1133─1145.
Cao, Y., Fu, Z.D., Wang, F., Liu, H.Y. & Han, R. (2005). Anti-Angiogenic Activity Of
Resveratrol, A Natural Compound From Medicinal Plants. Journal of Asian
natural products research 7(3): 205─213.
Choi, W.C., Kwon, S.P. & Park, S.J. (2009). Process for Preparation of Rhus Verniciflua
Extracts Having Excellent Anti-Cancer Activity and Anti-Cancer Pharmaceutical
Composition Containing The Same. Google Patents.
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an overview. International Journal of Pharmaceutical Sciences and Drug
Research 4(1): 1─9.
Cragg, G. M. & Newman, D. J. (2005). Plants as a Source of Anti-Cancer Agents. Journal
of ethnopharmacology 100(1): 72─79.
Elujoba, A. A., Odeleye, O. & Ogunyemi, C. (2005). Review-Traditional medicine
development for medical and dental primary health care delivery system in
Africa.
Feng, Y., Wang, N., Zhu, M., Feng, Y., Li, H. & Tsao, S. (2011). Recent Progress on
Anticancer Candidates in Patents of Herbal Medicinal Products. Recent patents
on food, nutrition & agriculture 3(1): 30─48.
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Montesano, R. & Schweigerer, L. (1997). Flavonoids, Dietary-Derived
Inhibitors of Cell Proliferation And in Vitro Angiogenesis. Cancer Research
57(14): 2916─2921.
Gupta, V. K. & Sharma S. K. (2006). Plants as Natural Antioxidants. Nat. Prod. Rad 5(4):
326─334.
Gururaj, A. E., Belakavadi, M., Venkatesh, D. A., Marme, D. & Salimath, B. P. (2002).
Molecular Mechanisms of Anti-Angiogenic Effect of Curcumin. Biochem
Biophys Res Commun 297(4): 934─942.
Huang, S.S. & Zheng R.l. (2006). Rosmarinic Acid Inhibits Angiogenesis and Its
Mechanism of Action In Vitro. Cancer letters 239(2): 271─280.
Kang, S. W., Choi, J. S., Choi, Y. J., Bae, J. Y., Li, J., Kim, D. S., Kim, J. L., Shin, S. Y., Lee, Y.
J., Kwun, I. S. & Kang, Y. H. (2010). Licorice Isoliquiritigenin Dampens
Angiogenic Activity via Inhibition of Mapk-Responsive Signaling Pathways
Leading to Induction of Matrix Metalloproteinases. J Nutr Biochem 21(1):
55─65.
Kintzios, S. E. & Barberaki, M. G. (2004). Plants That Fight Cancer, CRC Press.
Newman, D. J., Cragg, G. M. & Snader, K. M. (2000). The Influence of Natural Products
upon Drug Discovery. Natural product reports 17(3): 215─234.
Oberlies, N. H. & Kroll, D. J. (2004). Camptothecin and Taxol: Historic Achievements in
Natural Products Research. J Nat Prod 67(2): 129─135.
Pal, S., Bhattacharjee A., Ali, A., Mandal, N. C., Mandal, S. C. & M. Pal (2014). Chronic
Inflammation And Cancer: Potential Chemoprevention Through Nuclear Factor
Kappa B and P53 Mutual Antagonism. J. Inflamm.(Lond.) 11: 23.
Pettit, G., Singh, S., Hamel, E., Lin, C. M., Alberts, D. & Garcia-Kendal, D. (1989).
Isolation and Structure of The Strong Cell Growth and Tubulin Inhibitor
Combretastatin A-4. Experientia 45(2): 209─211.
Pham-Huy, L. A., He, H. &Pham-Huy, C. (2008). Free Radicals, Antioxidants in Disease
and Health. International journal of biomedical science: IJBS 4(2): 89.
Sakarkar, D. & Deshmukh, V. (2011). Ethnopharmacological Review of Traditional
Medicinal Plants for Anticancer Activity. International Journal of PharmTech
Research 3(1): 298─308.
69
Su, S.J., Yeh, T.M. , Chuang, W.J., Ho, C.L., Chang, K.L.,. Cheng, H.L, Liu, H.S., Cheng,
H.L., Hsu P.Y. & Chow, N.H. (2005). The Novel Targets for Anti-Angiogenesis of
Genistein on Human Cancer Cells. Biochemical pharmacology 69(2): 307─318.
Tan, W.F., Lin, L.P., Li, M.H., . Zhang, Y.X , Tong, Y.H., Xiao, D. & Ding, J. (2003).
Quercetin, a Dietary-Derived Flavonoid, Possesses Antiangiogenic Potential.
European journal of pharmacology 459(2): 255─262.
70
PEMBENTANGAN
POSTER
71
SARINGAN FITOKIMIA DAN AKTIVITI ANTI-RADANG TUMBUHAN
UBATAN TERPILIH WARISAN ORANG ASLI SEMELAI, POS ISKANDAR,
BERA, PAHANG
MA Adiana, H Nuziah, J Fadzureena, M Nik Musaadah, Y Norshakila, AL Tan, B
Intan Nurulhani, AM Nuraini, B Hani Idayu, SK Ling, HF Lim, MA Rasadah, & H
Norini
Institut Penyelidikan Perhutanan Malaysia (FRIM), 52109, Kepong, Selangor
Tel: 03-62797366
Faks: 03-62729805
ABSTRAK
Sebanyak 10 spesies tumbuhan ubatan berasaskan pengetahuan tradisi kaum
Semelai di Pos Iskandar, Bera, Pahang telah disaring untuk kandungan fitokimia
dan aktiviti anti-radang. Di dalam saringan fitokimia, 20 bahagian tumbuhan
telah menunjukkan kandungan saponin, 10 bahagian terdapat kehadiran
flavonoid, 21 bahagian menunjukkan kehadiran tannin terhidrolisis atau
terkondensasi, 11 bahagian menunjukkan kehadiran triterpenoid dan 15
bahagian menunjukkan kehadiran steroid. Aktiviti anti-radang atau dikenali
anti-inflamasi dilaksanakan melalui penentuan nilai perencatan enzim iaitu
enzim lipoksigenase dan enzim xanthine oksidase serta pembengkakan telinga
mencit yang diaruh dengan TPA (12-O-tetradekanoilforbol-13-asetat).
Perencatan aktiviti enzim tersebut boleh mengurangkan kesan buruk akibat
keradangan. Profil bioaktiviti dan fitokimia tumbuhan terpilih ini akan menjadi
asas kajian lanjutan R&D kearah pembangunan produk dari hasilan semula jadi.
Kata kunci: Fitokimia, anti-radang, pengetahuan tradisi, Semelai
PENGENALAN
Pendokumentasian pengetahuan tradisi adalah penting kerana maklumat
pengetahuan tradisi adalah bernilai tinggi dan merupakan sumber yang
penting. Melalui pendokumentasian yang sistematik, pengetahuan yang telah
lama diamalkan akan diperkasakan dan menjadi asas kepada penyelidikan dan
pembangunan tanpa mengabaikan hak harta intelek komuniti kaum yang
terlibat. Dokumentasi pengetahuan tradisi juga menyumbang kepada
pengenalpastian potensi baru sesuatu spesies tumbuhan.
Matlamat utama kajian ini adalah untuk mengenalpasti kandungan kimia
dan aktiviti anti-radang dari 10 spesies tumbuhan ubatan berasaskan
72
pengetahuan tradisi kaum Semelai di Pos Iskandar, Bera, Pahang. Kaedah
penyaringan fitokimia telah digunakan untuk mengenalpasti metabolit
sekunder yang dihasilkan oleh tumbuhan. Anti-radang atau dikenali antiinflamasi dilaksanakan melalui penentuan nilai perencatan enzim iaitu enzim
lipoksigenase dan enzim xanthine oksidase serta pembengkakan telinga mencit
yang diaruh dengan TPA. Perencatan aktiviti enzim tersebut boleh
mengurangkan kesan buruk akibat keradangan. Profil bioaktiviti dan fitokimia
tumbuhan terpilih ini akan menjadi asas kajian lanjutan R&D ke arah
pembangunan produk dari hasilan semula jadi.
BAHAN DAN KAEDAH
Penyaringan fitokimia
Flavonoid: Larutan klorofom dikeringkan dan kemudian diekstrak dengan
kombinasi ether dan larutan ammonia.
Saponin: Larutan methanol dan air digoncang untuk mengesan kehadiran buih.
(Simes 1995)
Tannin: Larutan methanol diuji dengan larutan ferum klorida.
Triterpenoid/steroid: larutan klorofom diuji dengan reagen LB (LiebermanBurchard).
Asai Xanthine Oksidase (XO):
Ekstrak sampel dilarutkan menggunakan 100% DMSO pada kepekatan 20
mg/mL. 130 µL larutan penimbal kalium fosfat 0.05 M (pH 7.5), 10 µL enzim XO
dipipet masuk ke dalam plat mikrotiter 96-telaga sebelum dieram selama 15
minit pada suhu 25°C. Selepas itu, tindakbalas enzim diaruh dengan
penambahan 100 µL substrat dan dieram lagi selama 10 minit. Aktiviti
perencatan enzim diukur menggunakan spektrofotometer pada 295 nm.
Allopurinol digunakan sebagai kawalan positif. Ujian diulang sebanyak tiga kali.
Asai Lipoksigenase (LOX)
Kaedah ujian adalah berdasarkan Azhar (2004) dengan sedikit modifikasi.
Sampel ditimbang dan dilarutkan menggunakan 100 % DMSO pada kepekatan
20 mg/mL. 160 µL larutan penimbal Na2H2PO4 (pH 8.0), berkepekatan 100 mM,
10 µL sampel ujian dan 20 µL larutan lipoksigenase kacang soya jenis 1B dipipet
masuk ke dalam mikroplat 96-telaga sebelum dieram selama 15 minit pada
73
suhu 25°C. Tindakbalas enzim kemudiannya dimulakan dengan penambahan
asid natrium linoleik (substrat) sebanyak 10 µL sebelum pengeraman dilakukan
sekali lagi selama 10 minit pada suhu 25°C. Aktiviti perencatan lipoksigenase
diukur dengan menggunakan spektrofotometer pada panjang gelombang 234
nm. Nordihidroguairetik asid (NDGA) digunakan sebagai kawalan positif. Ujian
diulang sebanyak tiga kali.
Asai Pembengkakan Telinga Mencit
Kajian ke atas model haiwan yang biasa digunakan untuk melihat aktiviti
antiradang bagi sampel yang diuji adalah dengan menggunakan aplikasi topikal
TPA (12-0-tetradekanoilforbol-13-asetat) untuk mengaruh bengkak atau
edema. Dalam kajian ini, aruhan bengkak pada telinga mencit dilakukan
dengan menyapu larutan TPA pada kedua-dua belah telinga mencit 40 minit
selepas larutan ekstrak/sampel disapu pada telinga kanan manakala aseton
disapu pada sebelah kiri sebagai kawalan. Selepas 6 jam pemberian TPA,
mencit akan dibunuh dan ditebuk kedua belah telinga pada diameter 7 mm
dan ditimbang. Kesan perencatan bagi setiap sample dikira dengan
membandingkan perbezaan berat antara telinga kiri dan kanan.
Sejumlah 22 bahagian daripada 10 spesies terpilih memberikan keputusan
positif terhadap ujian fitokimia (Jadual 1). Manakala, 20 bahagian tumbuhan
telah menunjukkan kandungan saponin, 10 bahagian terdapat kehadiran
flavonoid, 21 bahagian menunjukkan kehadiran tannin terhidrolisis atau
terkondensasi, 11 bahagian menunjukkan kehadiran triterpenoid dan 15
bahagian menunjukkan kehadiran steroid. Secara keseluruhan, spesies TD
08/10 dan TD 10/10 mempunyai kandungan flavonoid yang tinggi.
74
Jadual 1. Kehadiran kumpulan sebatian kimia. Sa=Saponin, Fla=Flavonoid,
Ta=Tannin, Tri= Triterpenoid, dan Ste= Steroid
Kod spesies Bahagian
tumbuhan
Sa
Fla
Ta
Tri
Ste
1
TD 01/10
daun
1+
0
C2
3+
3+
batang
1+
0
C2
2+
2+
rizom
1+
1+
C1
1+
0
2
TD 02/10
daun
2+
2+
C1
1+
2+
batang
2+
0
C1
0
0
dahan
3+
2+
C2
0
1+
3
TD 03/10
daun
1+
0
C2
3+
3+
batang
2+
1+
C1
0
0
4
TD 04/10
daun
2+
0
H2
3+
3+
batang
1+
0
H2
0
1+
5
TD 05/10
daun
1+
0
C2
3+
2+
6
TD 06/10
daun
1+
0
C3
3+
3+
batang
0
0
H1
1+
1+
dahan
0
2+
H2
0
1+
akar
0
0
0
0
0
7
TD 07/10
daun
0
0
C1
3+
3+
batang
0
2+
H1
0
0
8
TD 08/10
daun
1+
0
H2
0
3+
batang
1+
3+
C1
1+
0
9
TD 09/10
batang
3+
1+
0
0
0
kulit luar
1+
1+
C1
0
0
10
TD 10/10
daun
0
0
C2
0
1+
batang
1+
1+
H3
0
1+
Perencatan Aktiviti Enzim Xanthine Okidase, Lipoksigenase dan Asai
Pembengkakan Telinga Mencit
Hasil kajian anti-radang mendapati spesies TD 04/10 memberikan nilai
perencatan yang lebih tinggi terhadap enzim LOX iaitu bahagian daun (95.02 ±
4.76%) dan batang (84.92 ± 1.50%) manakala sampel TD 09/10 pula
mencatatkan nilai perencatan yang tinggi untuk bahagian kulit luar (86.18 ±
4.39%). Bahagian batang spesies TD 04/10 turut memberikan nilai perencatan
xanthine oksidase sebanyak 86.30% manakala bagi asai pembengkakan telinga
mencit pula bacaan sebanyak 29%. Nilai perencatan yang tinggi untuk enzim
xanthine oksidase juga direkodkan untuk spesies TD 08/10 dan TD 02/10.
Aktiviti anti-radang untuk spesies TD 07/10 dan TD 10/10 adalah sederhana
dan rendah.
75
Jadual 2. Aktiviti perencatan dari pelbagai bahagian tumbuhan untuk 10
spesies
Kod
Bahagian
Aktiviti anti-radang
tumbuhan
LOX (%)a
XO (%)a
TPA (%)
1
TD 01/10
daun
20.17± 4.44 41.79 ± 6.11
43%
batang
NA
24.74± 1.75
2%
rizom
10.63±7.97
56.42± 2.67
26%
2
TD 02/10
daun
50.33± 1.80
78.66± 5.32
41%
batang
42.81±2.12
57.75 ± 2.07
34%
dahan
62.04± 1.22 77.92 ± 4.06
26%
3
TD 03/10
daun
NA
63.20 ± 0.69
34%
batang
72.3± 6.28
61.40 ± 7.96
33%
4
TD 04/10
daun
95.02± 4.76
ND
26%
batang
84.92± 1.50 86.30 ± 5.08
29%
5
TD 05/10
daun
28.23±2.33
84.98 ± 7.21
49%
6
TD 06/10
daun
12.78± 6.44 41.47 ± 5.88
14%
batang
7.49±5.92
52.88 ± 7.07
7%
dahan
83.47± 5.33 83.18 ± 2.27
17%
akar
14.28± 6.36 46.97 ± 8.11
14%
7
TD 07/10
daun
10.55±5.86
10.35 ± 3.45
51%
batang
55.33±6.33
68.46 ± 1.81
37%
8
TD 08/10
daun
77.83±
74.33 ±
16%
14.33
batang
75.91± 4.67 81.40 ± 9.22
40%
9
TD 09/10
batang
12.12±3.03
NA
40%
Kulit luar
86.18±4.39
53.72±2.26
27%
10
TD 10/10
daun
27.33±1.99
27.13±3.79
34%
batang
33.27±1.28
50.44±3.15
26%
a
Semua nilai adalah min ± S.E.M., kepekatan akhir adalah 100 µg/mL.
Nota: H- Tinggi (71-100%) M-Sederhana (41-70%) L-rendah (0-40%) ND- Tidak
ditentukan
RUMUSAN
Secara keseluruhannya, spesies yang digunakan oleh kaum Semelai di Pos
Iskandar, Bera, Pahang mempunyai pelbagai kandungan kimia dan aktiviti antiinflamasi. Flavonoid, triterpenoid, saponin dan tannin dikesan hadir dalam
spesies yang digunakan.
76
RUJUKAN
Azhar, U.H., Abdul, M., Itrat, A., Sher Bahadar, K., Ezaz, A., Sarfraz, A.N., &
Muhammad, I. (2004). Enzymes Inhibiting Lignin from Vitex negundo.
Chemical and Pharmaceutical Bulletin: 52(11): 1269─1272.
Simes, J.J.H., Tracey J.G.,Webb, I.J. & Dunstaon, W.J. (1995). Australian
Phytochemicals Survey III. Saponins in the Eastern Australian Flowering
Plants. Bulletin No. 281., CSIRO, Melbourne Australia.
77
KANDUNGAN FITOKIMIA MAS COTEK BERLAINAN AKSESI
J Samsiah, D Thiyagu2, MS Rosalizan1, M Osman1, S Noor Safuraa1 & B Siti
Salwah3
1
Pusat Penyelidikan Hortikultur, Ibu Pejabat MARDI, Persiaran MARDI-UPM,
43400 Serdang, Selangor, Malaysia
2
Pusat Penyelidikan Padi dan Beras, MARDI Seberang Perai, Jalan Paya
Keladi/Pinang Tunggal, 13200 Kepala Batas, Pulau Pinang, Malaysia
3
Pusat Penyelidikan Padi dan Beras, Ibu Pejabat MARDI, Persiaran MARDI-UPM,
43400 Serdang, Selangor, Malaysia
Tel: 03-8953 6192
Faks: 03-8953 6270 E-mel: [email protected]
ABSTRAK
Mas cotek (Ficus deltoidea) digolongkan dalam famili Moraceae. Pokok ini
mempunyai pelbagai jenis bentuk daun, terdapat bentuk daun yang elips,
obovate atau spatulate. Daun berbentuk spatulate paling banyak ditanam dan
dijual. Daun-daun ini biasanya berwarna hijau, tebal dan keras dan hampir
berair. Ciri daun mas cotek adalah daun yang mempunyai bintik berwarna
emas pada permukaan daun. Daun mas cotek juga dikategorikan kepada
spesies jantan dan betina. Perbezaan daun mas cotek jantan dan betina adalah
berdasarkan bentuk daun dan warna bintik di bahagian bawah daun. Daun
pokok ini dikatakan mampu merawat pelbagai jenis penyakit. Kajian ini
dilaksanakan untuk menentukan kandungan vitexin (kandungan aktif) dalam
daun mas cotek berlainan aksesi. Kandungan aktif dikenal pasti dengan
menggunakan peralatan kromatografi cecair berprestasi tinggi (KCPT/HPLC).
Sepuluh aksesi mas cotek telah dianalisis dan dua aksesi, V10 dan V11
menunjukkan kandungan vitexin yang tinggi iaitu masing-masing pada 44.14
dan 20.46 µg/ml.
Kata kunci: mas cotek, aksesi, bentuk daun, vitexin
PENGENALAN
Mas cotek (Ficus deltoidea) dari famili Moraceae adalah pokok renek dengan
akar serabut yang biasanya bermula sebagai epifit. Pokok ini biasanya dilihat
sebagai pokok renek epifit yang melata pada pokok besar. Bentuk daun pokok
ini sangat berbeza-beza biarpun tergolong dalam genus yang sama. Bentuknya
sama ada eliptikal, lanseolat (seperti pisau pembedah) kepada obovat atau
spatulat. Mat-Salleh & Latiff (2002) menyatakan Ficus deltoidea mempunyai
enam varieti iaitu F. deltoidea var. angustifolia, F. deltoidea var. bilobata, F.
78
deltoidea var. trengganuensis, F. deltoidea var. kunstleri, F. deltoidea var.
intermedia dan F. deltoidea var. motleyana. Varieti-varieti ini dibeza
berdasarkan kelebaran buah, tulang daun (bercabang atau sebaliknya), bentuk
daun apeks (membulat, trunkat atau bentuk lain) serta saiz daun. F. deltoidea
var. deltoidea pula telah dilaporkan oleh Zunoliza et al. (2009). Sebanyak 30
aksesi mas cotek telah dikenal pasti berdasarkan morfologi bagi daun, batang
dan tabiat pertumbuhannya (Musa 2005).
Ekstrak daun mas cotek menunjukkan aktiviti anti inflammatori
(Zunoliza et al. 2009). Dalam perubatan tradisional, daun ini digunakan oleh
wanita sebagai rawatan selepas bersalin. Ia juga dipercayai dapat membantu
peredaran darah dan meningkatkan tenaga serta membantu merawat penyakit
berkaitan dengan gangguan haid (Burkill & Haniff 1930; Fasihuddin et al. 2002).
Padua et al. (1999) pula melaporkan daun kering mas cotek biasanya
digunakan oleh pengamal perubatan tradisional sebagai tonik selepas bersalin,
anti-hipertensi, anti-diabetik dan mengubat keputihan. Choo et al. (2012) telah
melaporkan kajian in vivo sebatian vitexin dan isovitexin daripada daun mas
cotek untuk perencatan -glucosidase.
BAHAN DAN KAEDAH
Bahan Tanaman
Sepuluh aksesi sampel mas cotek (MC) telah ditanam dan dikutip pada tiga
masa tuaian yang berbeza. Tuaian dilaksanakan pada 15 inci dari pucuk.
Sampel segar dikeringkan menggunakan pengering solar selama seminggu dan
kemudian dikisar untuk analisis kandungan sebatian vitexin.
Analisis Kandungan Vitexin Dalam 10 Mas Cotek Berlainan Aksesi
2.0 g daun kering yang telah dikisar diekstrak menggunakan pengekstrakan
soxhlet dengan pelarut metanol. Pelarut metanol ini kemudian dikeringkan
daripada sampel menggunakan penyejat vakum (rotary evaporator). Ekstrak
mentah dilakukan pencairan dengan 10 ml metanol (gred HPLC). Kemudian 0.1
ml daripada 10 ml ekstrak dilakukan pencairan lagi dengan 0.9 ml metanol gred
HPLC. Sampel yang dicairkan ini ditapis menggunakan picagari dengan penapis
0.45 µm sebelum dianalisis menggunakan HPLC. Setiap pengekstrakan
dilakukan dalam empat replikasi.
Sampel dipisahkan menggunakan kaedah gradient. Kaedah penentuan
vitexin adalah kaedah yang telah diubahsuai daripada yang telah dilaporkan
oleh Zunoliza et al. (2009). Turus C8 (150 mm x 4.6) Purospher STAR RP-8e (5
79
µm) Lichrocart (Merck). Aliran pelarut, 1.0 ml/min dilakukan selama 25 minit.
Puncak bagi vitexin dikesan pada gelombang 254 nm. Pelarut bergerak yang
terdiri daripada 0.1% H3PO4 dalam ACN (A) dan 0.05% H3PO4 dalam air yang
diionkan (B). Program bagi pelarut bergerak adalah seperti berikut:
Masa (minit)
6.0
11.0
14.5
25.0
Aliran (ml/min)
1.0
1.0
1.0
1.0
1.0
A (%)
10
15
25
35
10
B (%)
90
85
75
65
90
Standard vitexin telah ditemui pada masa penahanan, Rt = 15.422 min.
Persamaan bagi lengkuk kalibrasi bagi standard vitexin pada kepekatan 0.0325,
0.0650, 0.0975 dan 0.1300 mg/ml adalah Y = 6372X – 40165 x 105 dengan R2 =
0.994.
Sepuluh aksesi MC yang ditanam di Bachok, Kelantan telah dituai pada tiga
masa tuaian berbeza. Semua sampel ini telah dianalisa menggunakan HPLC dan
menunjukkan data kandungan vitexin yang baik. Sebatian vitexin (Rajah 1)
telah digunakan sebagai sebatian standard dalam MC. Perbezaan bentuk daun
MC ditunjukkan dalam Rajah 2.
OH
HO
HO
OH
O
HO
OH
O
OH
O
Rajah 1. Struktur kimia bagi sebatian utama MC
80
V1
V4
V 11
V 14
V6
V 19
V 10
V 23
V 28
V 39
Rajah 2. Perbezaan bentuk daun mas cotek
Nilai peratus ekstrak mentah semua aksesi menunjukkan nilai melebihi
10% dan hanya aksesi V1, V6, V11, V23 dan V36 menunjukkan peratus melebihi
nilai 25%. Semua aksesi kecuali VI, menunjukkan corak yang sama iaitu nilai
peratus berkurang pada tuaian kali kedua dan meningkat semula bagi tuaian
ketiga. Corak yang sama juga ditunjukkan oleh V4, V10, V11, V14, V19, V23 dan
V28 untuk kandungan vitexin. Daripada semua varieti, hanya V10 dan V11 yang
memberikan nilai kandungan vitexin melebihi 10 µg/ml pada tuaian ketiga.
Nilai peratus ekstrak mentah dan kandungan vitexin bagi sepuluh aksesi daun
MC yang berlainan telah diilustrasikan masing-masing dalam graf bar dan Rajah
3.
Rajah 3. Nilai peratus ekstrak mentah daun mas cotek berlainan aksesi
81
Jadual 1. Kandungan vitexin dalam aksesi berlainan daun mas cotek
Aksesi
V1
V4
V6
V10
V11
V14
V19
V23
V28
V39
Tuaian pertama
(µg/ml)
3.6650
5.0100
2.3338
20.4572
44.1429
4.1802
4.2593
9.4406
4.3228
6.0560
Tuaian kedua Tuaian ketiga
(µg/ml)
(µg/ml)
5.2235
5.5729
1.8567
2.9464
3.9667
4.9244
6.5406
12.1605
21.8074
40.1726
2.4409
3.4023
4.0829
4.6634
3.4575
7.4782
2.3226
2.6882
7.5688
6.2472
RUMUSAN
Sebanyak sepuluh aksesi mas cotek yang dituai pada tiga masa tuaian yang
berbeza telah dianalisis. Aksesi V1 merupakan aksesi terbaik yang
menunjukkan jumlah kandungan vitexin paling tinggi dengan nilai 44.14 µg/ml.
PENGHARGAAN
Penulis sangat menghargai Pejabat Pembangunan Herba (HDO) dan
Kementerian Pertanian dan Industri Asas tani untuk bantuan kewangan dengan
nombor vot K/G00N1210.
RUJUKAN
Burkill, I.H. & Haniff, M. (1930). Malay Village Medicine. Garden’s Bulletin 6(2):
67─332.
Choo, C.Y., Sulong, N.Y., Man, F. & Wong, T.W. (2012). Vitexin and Isovitexin
from the Leaves of Ficus deltoidea with In-Vivo -Glucosidase
Inhibition. Journal of Ethnopharmacology 142: 776–781.
Fasihuddin, B.A. & Din, L.B. (2002). Medicinal Plants Used by Ethnic Groups in
Sabah. Proceedings of French Malaysian-Symposium on Natural
Products. Universiti Malaya Kuala Lumpur. 85pp.
82
Mat-Salleh, K. & Latiff, M.A. (2002). Tumbuhan Ubatan Malaysia. Universiti
Kebangsaan Malaysia, Bangi. 184pp.
Musa, Y. (2005). Kepelbagaian Morfologi dan Agronomi Beberapa Aksesi Emas
Cotek yang Terdapat di Kelantan dan Terengganu. Buletin Teknologi
Tanaman 2 : 35─48.
Padua, L.S., Bunyapraphatsara, N. & Lemmens, R.H.M.J. (1999). Medicinal and
Poisonous Plants 1, Plant resources of South-East Asia No. 12 (1).
Bogor: Prosea Foundation Bogor.
Zunoliza, A., Khalid, H., Zhari, I. & Rasadah, M.A. (2009). Anti-Inflammatory
Activity of Standardized Extracts of Leaves of Three Varieties of Ficus
deltoidea. International Journal of Pharmaceutical and Clinical
Research 1(3): 100─105.
83
PENENTUAN JULAT EURYCOMANONE DALAM PRODUK TONGKAT ALI
MELALUI KAEDAH KROMATOGRAFI CECAIR ULTRAPRESTASI (UPLC)
A Mohd Radzi, SK Ling, L Abd Rashid & MT Nurzalikha
Tel: 03-6279 7335
ABSTRAK
Hasil kajian ini adalah berdasarkan penemuan kandungan eurycomanone di
dalam ekstrak dan produk tongkat ali melalui analisa UPLC. Analisa dijalankan
ke atas lebih 94 sampel yang diterima daripada 27 syarikat pengeluar herba,
agensi kerajaan dan orang perseorangan sejak tahun 2010 sehingga Disember
2014. Di antara sampel yang diterima adalah bahan mentah akar, daun, batang
dan buah tongkat ali, manakala yang lain adalah ekstrak tongkat ali, kapsul,
kopi campuran dan kultur tisu tongkat ali. Hasil analisa yang dijalankan ke atas
40 sampel akar tongkat ali mendapati 34 daripada sampel akar tersebut
mengandungi paras eurycomanone yang dapat dikesan. Nilai purata peratusan
kandungan eurycomanone dalam sampel akar yang dianalisa adalah 2.08%.
Nilai terendah yang dapat dikesan adalah 0.50% manakala yang tertinggi
adalah 4.36%. Purata eurycomanone yang dikesan di dalam esktrak dan kapsul,
masing-masing adalah 1.95%, dan 0.17%. NIlai purata eurymanone di dalam
daun tongkat ali adalah 0.41%, manakala satu daripada 3 sampel buah tongkat
ali yang diterima mengandungi peratusan kandungan 0.52%. Tujuan analisa
data ini dijalankan adalah untuk mencari julat eurycomanone terutamanya di
dalam produk tongkat ali. Oleh itu perkhidmatan ujian analisa eurycomanone
telah ditawarkan untuk membantu usahawan herba bagi mengelakkan
penyelewengan bahan mentah tongkat ali yang mereka perolehi, dan
bukannya untuk menentukan produk yang dihasilkan.
Kata kunci: julat, eurycomanone, Eurycoma longifolia, UPLC
PENGENALAN
Tongkat ali terkenal sebagai minuman herba yang paling popular di kalangan
pengusaha industri herba. Terdapat pelbagai produk minuman seperti kopi
campuran, teh tongkat ali dan juga minuman tin yang dicampurkan dengan
ekstrak tongkat ali. Hampir kesemua produk di pasaran dikatakan mempunyai
khasiat yang boleh meningkatkan tenaga. Berdasarkan laporan sebelum ini,
ekstrak tongkat ali mengandungi beberapa sebatian kimia dan eurycomanone
84
merupakan satu daripadanya (Zhari et al. 1999). Beberapa kajian yang
dijalankan oleh penyelidik melapurkan eurycomanone adalah bahan aktif bagi
aktiviti anti-malaria dan anti-ulcer (Chan et al. 2004). Eurycomanone turut
dijadikan sebatian penanda di dalam ekstrak organik dan akuas pada larutan
ekstrak piawai tongkat ali (Mohd Radzi et al. 2004). Analisa kandungan
eurycomanone telah dijalankan di Makmal Fitokimia (MF), Bahagian Hasilan
Semulajadi sejak tahun 2006. Analisa dijalankan dengan menggunakan
peralatan kromatografi cecair prestasi tinggi (HPLC) sehingga tahun 2009.
Bermula tahun 2010 hingga kini analisa dijalankan dengan menggunakan
peralatan kromatografi cecair ultraprestasi (UPLC).
Sebanyak lebih 27 syarikat dan agensi kerajaan dan swasta telah
menggunakan perkhidmatan dan kaedah yang telah dibangunkan oleh FRIM
untuk analisa eurycomanone. Secara puratanya lebih daripada lima syarikat
dan agensi menghantar sampel untuk dianalisa pada setiap tahun. Sebanyak
lebih 40 sampel bahan mentah akar tongkat ali dan juga akar yang dipercayai
tongkat ali telah diterima dan telah diproses. Makmal Fitokimia juga menerima
produk tongkat ali yang telah diproses dalam bentuk ekstrak untuk
menentukan paras eurycomanone dalam produk yang dihasilkan. Selain
daripada sampel di atas, MF juga menerima sampel daun, buah, kultur tisu
tongkat ali dan juga sebuk kopi campuran untuk di analisa. Analisa serbuk kopi
campuran tidak akan dibincangkan kerana hasil kajiannya telah dilapurkan
sebelum ini (Mohd Radzi et al. 2008).
BAHAN DAN KAEDAH
Sampel untuk kajian ini adalah sampel yang diterima oleh Makmal Fitokimia,
sejak tahun 2010 hingga 2014 (Jadual 1).
Penyediaan Sampel
Sampel yang diterima adalah berbentuk serbuk akar, ekstrak, serbuk daun atau
buah tongkat ali. Kesemua serbuk bahan mentah yang diterima akan ditapis
bagi mendapatkan sebuk yang bersaiz 1-5mm. Hasil tapisan kemudiannya akan
dicampurkan dengan larutan akuas pada kadar 1 nisbah 10 iaitu 1 gram sampel
dengan 10 mililiter akuas. Larutan kemudiannya diekstrak mengggunakan
sonicator pada suhu 50oC selama 90 minit. Larutan ditapis dengan kertas turas
dan hasil larutan dikeringkan menggunakan rotary evaporator. Berat ekstrak
yang dihasilkan ditimbang dan kemudiannya ditambahkan semula larutan
akuas dengan kepekatan akhir larutan mestilah tidak melebihi 3 mg/ml.
Larutan ekstrak mesti ditapis terlebih dahulu menggunakan penapis bersaiz
0.45 µ sebelum analisa UPLC dijalankan.
85
Bahan Kimia
Bahan kimia yang digunakan adalah acetonitril gred HPLC (MERCK) dan asid
formik (0.1%) (Fluka Chemica) sebagai pelarut fasa gerak. Eurycomanone (P)
(Chromadex) digunakan sebagai bahan piawai.
Analisa Kimia
Analisa kimia dijalankan menggunakan UPLC dengan sistem kawalan Waters
Acquity, Photodiode Array Detector, dan analisa data menggunakan Software
Empower 2 dan juga microsoft Excell. Penilaian eurycomanone terhadap
sampel produk dijalankan dengan menggunakan graf kelinearan kalibirasi
kepekatan eurycomanone (Rajah 1).
Kelinearan (linearity)
Kelinearan adalah dinilai melalui lakaran satu graf penentukuran melengkung
terhadap julat kepekatan 1 – 128 ppm eurycomanone. Luas kemuncak (peak
area) sebatian kimia adalah dibandingkan dengan kepekatan larutan piawai
eurycomanone. Cerun (slope), pintasan-y dan pekali penentuan (r2) adalah
didapati daripada perkiraan analisa regresi linear.
Jadual 1. Penerimaan sampel tongkat ali di FRIM (2010 hingga 2014)
Tahun
Syarikat
Akar
Ekstrak
Kapsul
Buah
Daun
2010
2011
2012
2013
2014
JUMLAH
5
5
3
7
7
27
20
1
0
10
9
40
10
4
2
5
9
30
0
1
1
0
4
6
2
1
0
0
0
3
10
0
0
0
0
10
86
Lainlain
4
0
0
0
1
5
JUMLAH
46
7
3
15
23
94
Rajah 1. Contoh graf kalibrasi untuk pengiraan eurycomanone
Kandungan dan peratus eurycomanone dinilai berdasarkan luas kemuncak
(dalam bulatan di Rajah 2) dan juga nilai kepekatan ekstrak yang digunakan
untuk analisa UPLC. Peratusan paras eurycomanone yang dikesan di dalam
sampel tongkat ali ditunjukkan dalam Jadual 2.
Hasil analisa yang dijalankan mendapati 34 daripada 40 sampel akar
mengandungi paras eurycomanone yang dapat dikesan. Nilai purata peratusan
kandungan eurycomanone dalam sampel akar yang dianalisa adalah 2.08%.
Nilai terendah yang dapat dikesan adalah 0.50% manakala yang tertinggi
adalah 4.36%. Sejumlah 27 daripada 30 sampel ekstrak produk tongkat ali yang
diterima mengandungi eurycomanone dengan nilai purata peratus yang
dikesan adalah 1.95%. Nilai terendah yang dihasilkan adalah 0.54% manakala
nilai tertinggi adalah 3.80%. Terdapat juga produk kapsul daripada ekstrak
tongkat ali yang diterima untuk dianalisa. Daripada lima sampel yang dianalisa
didapati purata nilai eurycomanone adalah rendah iaitu 0.17% dengan nilai
terendah yang dikesan adalah 0.05% manakala yang tertinggi adalah 0.2%.
Terdapat juga sampel daun tongkat ali yang diterima iaitu sebanyak 10 sampel.
Hasil analisa mendapati eurycomanone juga boleh dikesan di dalam daun
tongkat ali dengan kaedah yang digunapakai. Nilai purata eurycomanone
dalam daun adalah 0.41% dengan nilai terendah adalah 0.13% manakala yang
yang tertinggi adalah 0.64%. Selain daripada sampel di atas, MF juga
menerima sampel buah tongkat ali, serbuk kopi campuran dan juga kultur tisu
untuk dianalisa. Hanya satu sampel daripada tiga sampel buah mengandungi
peratus eurycomanone iaitu 0.52%, manakala yang lain tidak dapat dikesan.
87
Rajah 2. Profile UPLC untuk analisa sampel
Jadual 2. Paras eurycomanone dalam sampel
Sampel
Purata (%)
Minima (%)
Akar
2.08
0.50
Ekstrak
1.95
0.54
Kapsul
0.17
0.05
Daun
0.41
0.13
Maksima (%)
4.36
3.80
0.20
0.64
Jumlah (n)
34
27
5
10
Jadual 3. Bilangan sampel yang dapat dikesan paras eurycomanone
Tahun
JUMLAH
JUMLAH DIKESAN
Peratus dikesan
2010
46
41
89.1
2011
7
6
85.7
2012
3
3
100
2013
15
13
86.7
2014
23
21
91.3
JUMLAH
94
84
89.4
RUMUSAN
Sebagai rumusan daripada hasil analisa yang dijalankan terhadap 94 sampel
yang diterima, hampir 90% iaitu 84 sampel mengandungi sebatian
eurycomanone (Jadual 3). Peratus eurycomanone terendah dalam sampel akar
dan ekstrak tongkat ali adalah sekitar 0.5%, mungkin disebabkan oleh akar
yang belum cukup matang. Oleh itu, kajian peratus eurycomanone di dalam
akar tongkat ali pada umur yang berlainan perlu dijalankan. Manakala kadar
yang rendah di dalam ekstrak berkemungkinan bahan mentah dan cara
pengekstrakan yang dijalankan oleh pengusaha masih belum mencapai tahap
optima. Peratus eurycomanone di dalam produk kapsul yang dikesan adalah
tidak melebihi 0.2%. Berdasarkan pemerhatian ini, kemungkinan terdapat
88
campuran serbuk tongkat ali yang dikisar halus di dalam kapsul, berdasarkan
terdapatnya mendakan yang tidak larut di dalam larutan ujikaji. Mendakan
tidak larut ini boleh membahayakan kesihatan pengguna jika kapsul ini
dimakan dalam kuantiti yang melebihi had yang dibenarkan. Daun tongkat ali
juga mempunyai kadar peratusan eyrucomanone yang dapat dikesan. Satu
kajian menyeluruh juga harus dijalankan bagi mengkaji kesesuaian daun
sebagai bahan produk sebagai alternatif pengunaan akar.
RUJUKAN
Chan K.L, Choo C.Y., Noor Ain A. & Zakiah I. (2004). Antimalarial and
Toxicological Studies of Eurycomanone from Eurycoma Longifolia Jack
and Its Synthesized Derivatives. 25─28 Mac 2004 Herbal Asia, Kuala
Lumpur.
Mohd Radzi A., Mohd Ilham A., Norijas A.A., Mohd Noh J., & Mohd Ghawas M.
(2004). Penentuan Kualiti Berdasarkan Kandungan Kimia dan Usia
Penanaman Tongkat Ali Secara Perladangan. Pp 279-285. dlm Chang, Y.
S., Vimala, S., Mazura, P. & Ong, B. K. (eds). Proceedings of the Seminar
on Medicinal and Aromatic Plants: Current Trends and Perspectives.
Forest Research Institute Malaysia (FRIM), Kuala Lumpur.
Mohd Radzi, A., Ong, B. K., Mohd Ilham, A. & Mohd Hafidz, H.A. (2008)
Pembangunan Kaedah Penilaian Kawalan Mutu Minuman Kopi Tongkat
Ali Berasaskan Penanda Kimia Eurycomanone. Pp 272─279. Dlm Chang,
Y. S., Mazura, P. & Nik Musaadah, M. (eds). Proceedings Plant, Health
and Man-Past, Present and Future, FRIM, Kepong.
Zhari, I., Norhayati, I. & Jaafar. L. (1999). Radix Eurycomas. Malaysian Herbal
Monograph Vol. 1.: 29–32.
89
PENILAIAN KANDUNGAN FITOKIMIA DAN AKTIVITI SITOTOKSITI
SPESIES TERPILIH TUMBUHAN UBATAN BERASASKAN PENGETAHUAN
TRADISIONAL SUBETNIK JAHAI/TEMIAR DI KAMPUNG AIR BANUN,
GERIK, PERAK
MA Adiana, S Rohana, H Nuziah, J Fadzureena, M Nik Musaadah, AL Tan, B
Intan Nurulhani, AM Nuraini, SK Ling, HF Lim, MA Rasadah & H Norini
Institut Penyelidikan Perhutanan Malaysia (FRIM), 52109, Kepong, Selangor
Tel: 03-6279 7366
Faks:03-6272 9805 E-mel: [email protected]
ABSTRAK
Sepuluh spesies terpilih tumbuhan ubatan berasaskan pengetahuan tradisional
sub etnik Jahai/Temiar di Kampung Air Banun, Gerik, Perak telah dinilai dari
aspek kandungan fitokimia dan aktiviti sitotoksisiti. Sebanyak 31 bahagian
tumbuhan telah dinilai. Di dalam saringan fitokimia, 15 bahagian telah
menunjukkan kandungan saponin, 12 bahagian terdapat kehadiran flavonoid,
20 bahagian menunjukkan kehadiran tannin terhidrolisis atau terkondensasi,
27 bahagian menunjukkan kehadiran triterpenoid manakala 12 bahagian
menunjukkan kehadiran steroid. Untuk penilaian sitotoksisiti, ekstrak metanol
telah diuji ke atas dua jenis titisan sel; iaitu WRL-68 (human liver cells) dan
Vero (African green monkey kidney). Sel telah didedahkan kepada ekstrak
selama 72 jam dan aktiviti sitotoksisiti telah diuji dengan asai sulphorhodamine
B (SRB). Sebanyak 19 ekstrak mempamerkan sitotoksisiti sederhana (IC50 5-250
μg/mL) ke atas titisan sel WRL-68 dan lapan ekstrak adalah lemah sitotoksik
(IC50 250-625 μg/mL). Mengenai kesan sitotoksisiti ke atas sel-sel Vero, satu
ekstrak didapati sangat sitotoksik (IC50 0.01-5 μg/mL). Ekstrak selebihnya
didapati sederhana sitotoksik (19 ekstrak) manakala tujuh ekstrak adalah
lemah sitotoksik. Hanya ekstrak yang kurang toksik akan dipilih untuk dikaji
dengan lebih mendalam.
Kata kunci: fitokimia, sitotoksisiti, pengetahuan tradisi
PENGENALAN
Persepsi bahawa penggunaan ubatan herba adalah dijamin selamat
bersandarkan fakta ia berasal dari sumber semula jadi adalah kurang tepat.
Produk yang berasaskan semula jadi tidak semestinya selamat. Penggunaan
turun-temurun juga tidak boleh dijadikan bukti bahawasanya ia tidak akan
memberi risiko dan membahayakan pengguna dengan kesan-kesan sampingan
90
(Haq 2004). Bahan persediaan herba, ekstrak dan sebatian fitokimia mungkin
mempunyai risiko yang sama seperti sebatian sintetik.
Untuk memastikan penyelidikan bidang hasilan semula jadi dijalankan
dengan lancar, adalah sangat penting untuk mengenal pasti ekstrak/sebatian
fitokimia yang diperlukan dengan kaedah yang cepat dan ringkas. Kaedah yang
cepat dan ringkas perlu bagi mengelakkan pembaziran pengunaan sumber
tumbuhan yang kurang berpotensi.
Penyelidikan yang dijalankan ini adalah bertujuan untuk mengkaji
kandungan fitokimia dan sitotoksisiti ke atas beberapa tumbuhan ubatan
berasaskan pengetahuan tradisi sub etnik Jahai/Temiar di Kampung Air Banun,
Gerik, Perak. Ujian sitotoksisiti in vitro dengan menggunakan sel kultur telah
digunakan di dalam ujian toksisiti awalan kerana ia adalah cepat, menjimatkan
kos dan tidak mempunyai implikasi etika. Selanjutnya, hanya ekstrak yang
kurang toksik terhadap sel normal akan dipilih untuk dikaji dengan lebih
mendalam.
BAHAN DAN KAEDAH
Sepuluh spesies tumbuhan ubatan terpilih berasaskan pengetahuan tradisional
sub etnik Jahai/Temiar di Kampung Air Banun, Gerik, Perak dikenal pasti oleh
Program Sumber Biologi, Institut Penyelidikan Perhutanan Malaysia (FRIM).
Sampel-sampel tumbuhan dikering, dikisar dan kemudian diekstrak dengan
metanol. Ekstrak metanol kemudian dituras dan disejatkan di bawah tekanan
tinggi untuk menghasilkan ekstrak metanol kering.
Penyaringan Fitokimia
Flavonoid: Larutan klorofom dikeringkan dan kemudian diekstrak dengan
kombinasi ether dan larutan ammonia,
Saponin: Larutan metanol dan air digoncang untuk mengesan kehadiran buih.
(Simes, 1995)
Tannin: Larutan metanol diuji dengan larutan ferum klorida
Triterpene/steroid: larutan klorofom diuji dengan reagen LB (LiebermanBurchard)
91
Saringan Sitotoksiti Titisan Sel Normal
Dua jenis titisan sel normal telah digunakan di dalam penyelidikan ini iaitu
WRL-68 (human liver cells) dan Vero (African green monkey kidney). Keduadua jenis titisan sel normal diperolehi daripada ATCC. Ia telah dikulturkan di
dalam Medium Dulbecco’s Modified Eagle medium (Sigma, USA) dan
dicampurkan dengan serum 5% foetal bovine (PAA Laboratories), amphotericin
B 1 % dan penisilin-streptomisin (PAA Laboratories) dan gentamycin 0.5% (PAA
Laboratories). Sel disimpan di dalam inkubator yang mengandungi 5%
CO2/95% air pada suhu 37°C.
Sel yang yang telah tumbuh pada kadar eksponential telah diletakkan
di dalam piring 96-well untuk mencapai jumlah 10,000 sel/telaga. Selepas 24
jam tempoh pengeraman, sel telah didedahkan dengan ekstrak tumbuhan
dengan kepekatan berjulat 0.1 hingga 1000 µg/ml untuk tempoh 72 jam. Untuk
tujuan perbandingan, Paclitaxel telah digunakan dan setiap eksperimen
dijalankan di dalam triplikat.
Selepas 72 jam tempoh inkubasi, petunjuk kadar ketoksikan ditentukan
dengan menggunakan asai sulphoramide B (SRB) (Skehan et al. 1989) dengan
sedikit modifikasi seperti yang dijelaskan oleh Voight (2005). Keputusan
dilaporkan dalam peratusan sel yang yang hidup dan kepekatan perencatan
median (MIC, IC50) yang ditentukan daripada keluk dos kepekatan (dose
response curve).
Saringan fitokimia telah dijalankan ke atas 10 spesies tumbuhan ubatan dan
hasil keputusan dirumuskan di dalam Jadual 1. Sebanyak 31 bahagian
tumbuhan telah dinilai. Di dalam saringan fitokimia, 15 bahagian telah
menunjukkan kandungan saponin, 12 bahagian terdapat kehadiran flavonoid,
20 bahagian menunjukkan kehadiran tannin terhidrolisis atau terkondensasi,
27 bahagian menunjukkan kehadiran triterpenoid manakala 12 bahagian
menunjukkan kehadiran steroid.
Keputusan ujian sitotoksisiti ditunjukkan di Jadual 2. Untuk penilaian
sitotoksisiti, ekstrak metanol telah diuji ke atas dua jenis titisan sel iaitu WRL68 dan Vero. Sel telah didedahkan kepada ekstrak selama 72 jam dan aktiviti
sitotoksisiti telah diuji menggunakan asai sulphorhodamine B (SRB). Sebanyak
19 ekstrak mempamerkan sitotoksisiti sederhana (IC50 5-250 μg/ml) ke atas
titisan sel WRL-68 dan lapan ekstrak adalah lemah sitotoksik (IC50 250-625
μg/ml). Mengenai kesan sitotoksisiti ke atas sel-sel Vero, satu ekstrak iaitu
bahagian daun spesies TD 07/11 didapati sangat sitotoksik (IC50 0.01-5 μg/ml).
92
Ekstrak selebihnya didapati sederhana sitotoksik (19 ekstrak) dan lemah
sitotoksik (tujuh ekstrak). Hanya ekstrak yang kurang toksik akan dipilih untuk
kajian lanjutan.
Jadual 1. Taburan sebatian fitokimia yang terdapat di dalam 10 spesies
tumbuhan herba berasaskan pengetahuan tradisi.
Kod spesies
Bahagian
tumbuhan
Sa
Fla
Ta
Tri
1
TD 01/11
batang (tua)
1+
1+
2+
batang (muda)
2+
1+
1+
2+
daun
2+
2+
2+
2+
2
TD 02/11
stem
1+
3+
2+
daun
1+
2+
3+
3+
akar
1+
2+
bunga
2+
2+
3
TD 03/11
stem
1+
1+
1+
daun
2+
3+
2+
buah
1+
1+
4
TD 04/11
stem
1+
2+
3+
3+
daun
1+
3+
akar
5
TD 05/11
stem
2+
2+
2+
daun
2+
6
TD 06/11
batang
1+
2+
daun
1+
1+
2+
2+
akar
1+
1+
7
TD 07/11
stem (muda)
1+
3+
stem (tua)
1+
3+
3+
daun
2+
2+
3+
8
TD 08/11
stem
3+
daun
batang (tua)
1+
1+
1+
3+
batang (muda)
3+
9
TD 09/11
stem
2+
2+
daun
2+
2+
3+
bunga
1+
1+
Rizom
1+
1+
10 TD 10/11
stem
2+
2+
2+
daun
NT
NT
NT
NT
* Sa=Saponin, Fla=Flavonoid, Ta=Tannin, Tri= Triterpene dan Ste=Steroid
93
Ste
H2+
C2+
C2+
C3+
C1+
C1+
C2+
C2+
C2+
C2+
2+
C2+
NT
Jadual 2. Kesan sitotoksiti 10 spesies tumbuhan herba berasaskan pengetahuan
tradisional sub etnik Jahai/Temiar di Kampung Air Banun, Gerik, Perak
Kod spesies
1.
TD 01/11
2.
TD 02/11
3.
TD 03/11
4.
TD 04/11
5.
TD 05/11
6.
TD 06/11
7.
TD 07/11
8.
TD 08/11
9.
TD 09/11
10.
TD 10/11
Bahagian tumbuhan
Sitotoksisiti (IC50) ug/ml
Vero
77.5± 2.3
93.3±5.1
382.6±16.6
415.8±5.5
83.5±0.2
338.3±14.3
78.6±2.8
73.5±3.4
75.4±4.2
337.9±2.9
NT
165.45±63.0
164.6±77.5
372.6±5.9
8.7±3.4
NT
NT
76.6±3.6
18.6±0.4
31.2±4.9
4.7±2.7
73.45±7.3
368.1±5.5
373.7±17.3
247.4±36.9
81.4±0.5
NT
82.1±2.9
76.2±0.7
77.4±2.2
76.3±1.1
batang (tua)
batang (muda)
daun
stem
daun
akar
bunga
stem
daun
buah
stem
daun
akar
stem
daun
batang
daun
akar
stem (muda)
stem (tua)
daun
stem
daun
batang (tua)
batang (muda)
stem
daun
bunga
rizom
stem
daun
NT: Tidak ditentukan
94
WRL-68
136.2±16.9
282.±21.1
428.0±18.7
510.6±17.5
114.6±5.3
211.5±26.4
98.8±5.2
62.5±2.5
83.1±6.0
375.7±9.6
NT
170.65±21.1
184.25±60.8
433.0±12.6
9.3±2.7
NT
NT
100.3±0.0
25.4±2.7
49.8±9.7
13.2±0.0
72.9±0.7
399.6±32.5
349.95±11.8
111.5±5.2
54.6±9.5
NT
383.2±43.8
73.7±2.4
79.8±1.1
71.2±5.8
RUMUSAN
Data yang diperolehi daripada kajian ini memberikan informasi yang berguna
untuk tujuan penyelidikan yang lebih mendalam ke atas spesies-spesies
tersebut yang mempunyai potensi biologi.
RUJUKAN
Haq, I. (2004). Safety of medicinal plants. Pakistan Journal of Medical Research.
43. 203─210.
Skehan P., Storeng R., Scuidero D., Monks A., McMahon J., Vistica D., Warren
J.T., Bokesch H., Kenney S. & Boyd M.R. (1990). New Calorimetric
cytotoxicity assay for anti-cancer-drug screening. Journal of the
National Cancer Institute. 82: 1107─1102.
Voight W. (2005). Sulfordamine B Assay and chemosensitivity. In: Blumenthal
R.D (Editor): Chemosensitivity. Vol 1: In vitro assays. Methods in
Molecular Medicine. Humana Press Inc. 39─48.
95
ASAS MOLEKUL AKTIVITI ANTI-MALARIA KURKUMIN
A Amatul Hamizah, S Suhaini, E Noor & MS Hasidah
Pusat Pengajian Biosains dan Bioteknologi, Fakulti Sains dan Teknologi,
Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia
Tel: 03-8921 5954
ABSTRAK
Kurkumin, sebatian bioaktif utama dalam kunyit (Curcuma longa) dilapor
mempamerkan pelbagai aktiviti biologi seperti kesan antikanser, antidiabetes,
antiinflamasi dan anti-malaria. Kajian ini dilakukan untuk memahami asas
molekul aktiviti anti-malaria kurkumin melalui eksperimen haiwan (in vivo) dan
kultur parasit (in vitro). Perlakuan secara terapeutik dan profilaksis
(intraperitoneum @ 30 mg/kg berat mencit) setiap satu menunjukkan bahawa
kurkumin dapat menindas perkembangan parasit dalam mencit terinfeksimalaria. Di samping itu, perlakuan kurkumin meningkatkan kemandirian
haiwan terinfeksi. Kurkumin juga mempamerkan aktiviti antiplasmodium (in
vitro) yang baik dan selektif (IC50=4.34±1.59 µM; SI=28.98). Analisis protein
hepar menunjukkan bahawa perlakuan kurkumin menyebabkan perencatan
enzim glikogen sintase kinase-3 beta (GSK3β). Penemuan ini menjelaskan asas
molekul aktiviti anti-malaria kurkumin dan merupakan bukti saintifik
penggunaan kurkumin dalam rawatan tradisional untuk malaria.
Kata kunci: kurkumin, anti-malaria, glikogen sintase kinase-3β
PENGENALAN
Kurkumin merupakan sebatian bioaktif daripada Curcuma longa (kunyit) yang
dilaporkan mempamerkan aktiviti farmakologi yang luas antaranya antikanser
(Kizhakkayil et al. 2012), antidiabetes (Sharma et al. 2006), antiinflamasi (Kohli
et al. 2005) dan anti-malaria (Mimche et al. 2011). Selain itu, kurkumin juga
telah digunakan sebagai terapi sampingan (adjunktif) bagi pesakit malaria
serebrum (Jain et al. 2013). GSK3 penting dalam mengawal sistem imun yang
melibatkan pelbagai komponen inat dan adaptif. GSK3β merupakan
pengawalatur transkripsi faktor ‘Nuclear Factor Kappa B’ (NF-kB) yang
berkaitan dengan kadar inflamasi khususnya pada peningkatan dan penurunan
aras pro- dan anti-inflamasi sitokin (Cortes-Vieyra et al. 2012). Menurut
Bustanji et al. (2009), kurkumin berpotensi bertindak sebagai perencat GSK3β
berdasarkan kajian simulasi docking dalam kajian in silico. Setakat ini,
96
penglibatan kurkumin dalam merencat pengaktifan GSK3 dalam kajian malaria
masih belum jelas.
Malaria adalah penyakit disebabkan oleh infeksi protozoa daripada
genus Plasmodium. Kes kematian yang tinggi disebabkan oleh peningkatan
bilangan strain parasit rintang drug anti-malaria lazim. Penularan kes malaria di
serata dunia disebabkan oleh kegagalan program pengawalan malaria secara
global kerana kebergantungan kepada penggunaan artemisinin sebagai drug
anti-malaria utama dan kemunculan strain parasit yang rintang terutama di
negara-negara Asia Tenggara (Dondorp et al. 2009). Di Malaysia, kemunculan
infeksi zoonosis berlaku akibat jangkitan P. knowlesi bermula daripada haiwan
primat, seperti beruk dan monyet dan kini menyerang manusia terutamanya di
kawasan pedalaman seperti di Sabah dan Sarawak. Bagi mengatasi penularan
ini, industri farmaseutikal giat menyaring dan membangunkan pelbagai drug
anti-malaria sama ada melalui pemencilan daripada sumber semulajadi
ataupun penghasilan drug anti-malaria secara sintetik. Kedua-dua drug antimalaria utama telah dibangunkan daripada produk tumbuhan iaitu daripada
Cinchona sp. dan Artemisia annua. Dalam penyelidikan ini, kurkumin dikaji bagi
memahami asas molekul aktiviti anti-malaria sebatian bioaktif ini,
memandangkan kurkumin juga berpotensi digunakan dalam terapi sampingan.
Penemuan daripada kajian ini diharap menyumbang kefahaman tentang asas
molekul dan mekanisme tindak balas kurkumin dalam infeksi malaria.
BAHAN DAN KAEDAH
Pengkulturan P. falciparum dan Pengasaian Aktiviti Antiplasmodium
Strain P. falciparum sensitif-klorokuin (3D7) diperoleh daripada Malaria
Research and Reference Reagent Resource Center (MR4), USA dan dikultur
berdasarkan kaedah Trager & Jensen (1976).
Pengkulturan Sel Chang dan Asai Sitotoksiti
Sel hepar Chang, diperoleh daripada American Type Culture Collection (ATCC)
dan dikultur dalam Dulbecco Modified Eagle Medium (DMEM) (Gibco, USA)
berdasarkan kaedah Mossmann (1983).
Ujian Anti-malaria Empat Hari Penekanan Terapeutik dan Profilaksis
Mencit jantan ICR (25±5 g; berusia 6-8 minggu) diperoleh daripada Kompleks
Rumah Haiwan, UKM. Kelulusan untuk kajian haiwan diperoleh daripada
97
Jawatankuasa Etika Haiwan Universiti Kebangsaan Malaysia (UKMAEC). Strain
Plasmodium berghei NK65 (klorokuin-sensitif) daripada Malaria Research and
Reference Reagent Resource Center (MR4, USA). Ujian penekanan terapeutik
dan profilaksis berdasarkan kaedah Peters (1975).
Penentuan Status Pemfosfatan Gsk3β Melalui Analisis Pemblotan
Western
Pengekstrakan protein daripada hepar mencit dilakukan mengikut kaedah Lee
et al. (2007). Kandungan protein sampel diukur menggunakan kaedah Bradford
(1976) dengan bovin serum albumin (BSA) sebagai piawai. Pemisahan protein
dalam gel elektoforesis berdasarkan kaedah Laemmi (1970).
Kurkumin Mempamerkan Aktiviti Anti-Plasmodium yang Baik dan
Selektif Terhadap Parasit Malaria
Hasil penentuan aktiviti antiplasmodium secara in vitro menunjukkan kurkumin
berupaya merencat pertumbuhan P. falciparum 3D7 dengan baik (4.34 ± 1.59
µM). Hasil kajian ini mempunyai persamaan dengan hasil kajian (Reddy et al.
2005) yang melibatkan strain P. falciparum yang lain.
Kurkumin Merencat Perkembangan Parasit dalam Mencit TerinfeksiMalaria
Pada dos 30 mg/kg berat tubuh, kurkumin menindas lebih 50–60% populasi P.
berghei NK65 melalui ujian terapeutik dan profilaksis dan mampu melanjutkan
tempoh hayat mencit terinfeksi-malaria. Hasil serupa telah dilaporkan oleh
(Reddy et al. 2005) tetapi melibatkan strain yang berbeza.
Kurkumin menyebabkan perencatan enzim GSK3β
Daripada analisis protein bagi ujian terapeutik dan profilaksis, didapati bahawa
kurkumin mempunyai ciri perencatan enzim GSK3 berbanding haiwan yang
tidak diberi rawatan (data tidak ditunjukkan). Walaupun sebelum ini, kurkumin
pernah dilaporkan mempunyai kesan perencatan terhadap molekul pengatur
sistem inflamasi sel (NF-kB), penemuan kami bahawa kurkumin menyebabkan
penindasan aktiviti GSK3 merupakan laporan buat pertama kali. Hasil kajian ini
selari dengan peranan GSK3 dalam mengawal pengaktifan NF-kB.
98
KESIMPULAN
Hasil kajian ini merupakan satu penemuan baru mengenai kefahaman asas
molekul aktiviti anti-malaria kurkumin yang berasaskan eksperimen pada
haiwan (in vivo) dan kultur parasit (in vitro).
PENGHARGAAN
Penyelidikan ini dibiayai oleh geran daripada Universiti Kebangsaan Malaysia
(UKM-ETP-2013-052).
RUJUKAN
Bradford, M.M. (1976). A Rapid and Sensitive Method for the Quantitation of
Microgram Quantities of Protein Utilizing the Principle of Protein-Dye
Binding. Analytical Biochemistry: 72(1─2): 248─254.
Bustanji, Y., Mutasem, O.T., Ihab, M.A., Mohamed, A.S.A., Mohammad, K.M. &
Hatim, S.A. (2009). Inhibition of Glycogen Synthase Kinase By
Curcumin: Investigation by Simulated Molecular Docking and
Subsequent In Vitro/In Vivo Evaluation. Journal of Enzyme Inhibition
and Medicinal Chemistry: 24 (3):771─778.
Cortés-Vieyra, R., Alejandro, B.P., Juan, J.V.A., Marcos, C.J., Finlay, B.B. &Víctor,
M. B.A. (2012). Role of Glycogen Synthase Kinase-3 Beta in The
Inflammatory Response Caused by Bacterial Pathogens. Journal of
Inflammation (Lond): 9 (1):23─23.
Dondorp, A.M., François, N., Poravuth, Y., Debashish, D., Aung, P.P., Joel, T.,
Khin, M.L., Frederic, A., Warunee, H. & Sue, J.L. (2009). Artemisinin
Resistance in Plasmodium Falciparum Malaria. New England Journal of
Medicine: 361 (5):455─467.
Jain, K., Sood, S. & Gowthamarajan, K. (2013). Modulation of Cerebral Malaria
by Curcumin as An Adjunctive Therapy. The Brazilian Journal of
Infectious Diseases: 17(5): 579─591.
Kizhakkayil, J., Faisal, T., Shahanas, C., Abdulkader, H., Mahendra, P.&
Sehamuddin, G. (2012). Glutathione Regulates Caspase-Dependent
Ceramide Production and Curcumin-Induced Apoptosis in Human
Leukemic Cells. Free Radical Biology and Medicine: 52 (9):1854─1864.
Kohli, K., Ali, J., Ansari, M.J.& Raheman, Z. (2005). Curcumin: A Natural
Antiinflammatory Agent. Indian Journal of Pharmacology: 37 (3):141.
99
Leammi, U.K. (1970). Cleavage of Structural Proteins During The Assembly of
The Head of Bacterial Phage T4. Nature: 227( 5259):680─685.
Lee, C. (2007). Protein Extraction from Mammalian Tissues. Methods in
Molecular Biology: 362: 385─389.
Mimche, P.N., Donatella, T. & Livia, V. (2011). The Plant-Based
Immunomodulator Curcumin as A Potential Candidate for The
Development of an Adjunctive Therapy for Cerebral Malaria. Malaria
Journal: 10 (1):10.
Mosmann, T. (1983). Rapid Colorimetric Assay for Cellular Growth and Survival:
Application to Proliferation and Cytotoxicity Assays. Journal of
Immunological Methods: 65(1): 55─63.
Peters, W. (1975). The Chemotherapy of Rodent malaria, xxii. The Value of
Drug-Resistant Strains of in Screening for Blood Schizontocidal Activity.
Annals of Tropical Medicine and Parasitology: 69 (2): 155─171.
Reddy, R.C., Vatsala, P.G., Keshamouni, V.G., Padmanaban, G., & Rangarajan,
P.N. (2005). Curcumin for Malaria Therapy. Biochemical and
Biophysical Research Communications: 326(2), 472─474.
Sharma, S., Kulkarni, S.K., & Chopra, K. (2006). Curcumin, The Active Principle
of Turmeric (Curcuma Longa), Ameliorates Diabetic Nephropathy in
Rats. Clinical and Experimental Pharmacology and Physiology: 33(10):
940─945.
Trager, W. & Jensen, J.B. (1976). Human Malaria Parasites in Continuous
Culture. Science: 193(4254): 673─675.
100
MEKANISME TINDAKAN SEBATIAN BIOAKTIF ANTI-MALARIA DALAM
PAKU PAKIS UBATAN, GLEICHENIA TRUNCATA
S Suhaini, E Noor & MS Hasidah
Pusat Pengajian Biosains & Bioteknologi, Fakulti Sains & Teknologi,
Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor
Tel: 03-8921 5954
ABSTRAK
Gleichenia truncata, sejenis paku pakis daripada famili Gleicheniaceae
digunakan di kalangan masyarakat pribumi Asia secara meluas termasuk di
Sabah sebagai rawatan tradisional bagi penyakit berkait-inflamasi seperti
demam dan ulser. Sebelum ini kami telah melaporkan asas saintifik aktiviti antimalaria yang hadir dalam ekstrak kasar G. truncata. Kajian ini ialah mengenai
aktiviti anti-malaria sebatian aktif G. truncata yakni metil-4-hidroksisinamat
(M4H). Hasil kajian infeksi malaria dalam haiwan menunjukkan perlakuan
secara intraperitoneum (IP) 30 mg/kg berat tubuh (BT) M4H menindas
perkembangan parasit P. berghei NK65 (kemopenindasan >60%) dalam
eritrosit. Kemandirian mencit terinfeksi- Plasmodium berghei yang diberi
perlakuan M4H adalah lebih tinggi berbanding kumpulan haiwan kawalan
terinfeksi tanpa perlakuan. Analisis Western enzim GSK3 dalam hepar semasa
infeksi P. berghei menunjukkan perlakuan 30 mg/kg BT M4H meningkatkan
pemfosfatan GSKβ berbanding kawalan. Hasil ini mencadangkan bahawa M4H
adalah satu sebatian aktif yang menyumbang kepada aktiviti anti-malaria dan
asas saintifik paku pakis G. truncata sebagai tumbuhan ubatan.
Kata kunci: Gleichenia truncata, anti-malaria, glikogen sintase kinase-3β
PENGENALAN
Penggunaan tumbuhan sebagai ubatan tradisional menjadi tunjang utama
penyelidik masa kini untuk mengeksploitasi nilai farmakologi daripada sumber
semula jadi. Gleichenia truncata (famili Gleicheniaceae) digunakan secara
meluas oleh masyarakat Asia untuk merawat penyakit berkait-inflamasi seperti
ulser dan demam (Ho et al. 2010). Kajian saintifik daripada makmal kami telah
melaporkan aktiviti anti-inflamasi atau anti-malaria ekstrak kasar metanol
tumbuhan ini dalam model kajian infeksi melioidosis dan malaria (Suhaini et al.
2015). Kajian penyaringan fitokimia G. truncata menunjukkan kehadiran
beberapa kelas sebatian yang pernah dilapor mempamer aktiviti anti-malaria
seperti flavonoid dan asid fenol (Chai et al. 2013).
101
Usaha penyelidikan berterusan dijalankan melalui eksploitasi fungsi
protein kinase yang berpotensi dijadikan sebagai sasaran drug anti-malaria.
Salah satu protein kinase yang diberi perhatian dalam kajian sasaran drug antimalaria adalah glikogen sintase kinase-3 (GSK3), iaitu kinase serin/treonin yang
berfungsi sebagai kunci utama dalam penyakit berkait-inflamasi penyakit
infeksi virus, bakteria serta parasit menjadikan kinase ini sebagai sasaran
berpotensi bagi drug anti-malaria (Dai et al. 2012). Perencatan aktiviti GSK3
dilihat mampu memberi perlindungan kepada hos kerana enzim ini
merangsang penghasilan sitokin pro-inflamasi secara berlebihan semasa infeksi
malaria (Dai et al. 2012). Perencat GSK3, LiCl dilaporkan berupaya merencat
pertumbuhan Plasmodium falciparum dan P. berghei masing-masing melalui
kajian in vitro dan in vivo (Marhalim et al. 2014; Nurul Aiezzah et al. 2010).
Justeru itu, perencatan aktiviti GSK3 bukan sahaja dilihat mampu
mengurangkan kesan patologi malaria dalam hos, malahan merencat
perkembangan parasit semasa infeksi malaria. Dalam kajian ini kesan
perlakuan metil-4-hidroksisinamat (M4H), iaitu sebatian bioaktif G. truncata
diuji menggunakan model haiwan malaria.
BAHAN DAN KAEDAH
Ujian Penindasan Empat Hari
Ujian penindasan empat hari dijalankan berdasarkan kaedah Peters (1975).
Alikuot parasit disuntik secara intraperitoneum (IP) kepada mencit dalam
setiap kumpulan kajian pada hari infeksi (H0). Selepas tiga jam infeksi,
kumpulan mencit perlakuan dos M4H manakala kumpulan haiwan kawalan
tanpa perlakuan diberikan larutan 0.9% salina secara IP selama empat hari
berturut-turut, iaitu bermula pada hari infeksi (H0) hingga hari ketiga (H3).
Peratus penindasan perkembangan parasit ditentukan melalui slaid calitan
nipis darah mencit pada hari keempat (H4) dan kemandirian haiwan
direkodkan sehingga hari ke-30 (H30).
Penentuan Status Pemfosfatan GSK3
Hepar yang diperoleh pada H4 ditimbang untuk tujuan pengekstrakan protein
berdasarkan kaedah Lee (2007). Protein jumlah ekstrak hepar ditentukan
mengikut kaedah Bradford (1976) untuk langkah pemisahan protein
menggunakan teknik SDS-PAGE (Laemmli 1970). Gel yang mengandungi protein
yang telah dipisahkan dipindahkan ke tangki pemblotan western (Towbin et al.
1979). Jalur protein imunoreaktif pada membran dikesan menggunakan sistem
pengesanan pendaflour kimia dan dipindahkan pada filem sinar-X.
102
Sebatian M4H menindas perkembangan parasit dalam mencit
terinfeksi-malaria
Hasil kajian in vivo dalam model infeksi-P. berghei NK65 (strain sensitifklorokuin) menunjukkan pemberian 30 mg/kg BT M4H merencat
perkembangan parasit P. berghei peringkat eritrosit dengan aktif
(kemopenindasan melebihi 60% pada H4 selepas infeksi) berdasarkan
pengkelasan Rasoanaivo et al. (2004). Pemberian 30 mg/kg BT M4H juga
dicerap mampu memanjangkan tempoh kemandirian haiwan terinfeksi-P.
berghei berbanding kumpulan kawalan tanpa perlakuan (data tidak
ditunjukkan). Pemerhatian ini mungkin berkait dengan pengurangan kesan
patologi malaria (Mota & Rodriguez 2002).
Sebatian M4H menyebabkan perencatan enzim GSK3β
Penilaian status pemfosfatan GSK3β hepar yang diperoleh daripada haiwan
terinfeksi diberi 30 mg/kg BT M4H menunjukkan peningkatan aras pemfosfatan
GSK3β berbanding kumpulan tanpa perlakuan (larutan 0.9% salina) (data tidak
ditunjukkan). M4H yang hadir dalam G. truncata berperanan merencat aktiviti
GSK3β hos semasa infeksi P. berghei yang seterusnya dapat mengurangkan
kesan patologi malaria dan memanjangkan hayat haiwan terinfeksi.
KESIMPULAN
Hasil kajian ini mencadangkan asas saintifik penggunaan tumbuhan ubatan G.
truncata secara tradisional dalam merawat penyakit berkait-inflamasi.
PENGHARGAAN
Penyelidikan ini dibiayai oleh geran daripada ScienceFund Kementerian Sains,
Teknologi dan Inovasi (MOSTI) (02─01─02─SF1248).
RUJUKAN
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Antibacterial, anti-glucosidase, and antioxidant activities of selected
highland ferns of Malaysia. Botanical Studies: 1─7.
Dai, M., Freeman, B., Shikani, H. J., Bruno, F. P., Collado, J. E., Macias, R.,
Reznik, S. E., Davies, P., Spray, D. C., Tanowitz, H. B., Weiss, L. M. &
Desruisseaux, M. S. (2012). Altered regulation of Akt signaling with
murine cerebral malaria, effects on long-term neuro-cognitive
function, restoration with lithium treatment. PLoS One: 1─15.
Ho, R., Teai, T., Bianchini, J.-P., Lafont, R. & Raharivelomanana, P. (2010).
Working with Ferns. Springer New York.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head
of bacteriophage T4. Nature: 680─685.
Lee, C. (2007). Protein extraction from mammalian tissues. Methods in Molecular
Biology: 385─389.
Marhalim, L., Embi, N. & Sidek, H.M. (2014). Synergistic effects of GSK3 and
p38 mapk inhibitors on growth of Plasmodium falciparum ex vivo.
Malaysian Applied Biology: 65─71.
Mota, M. M. & Rodriguez, A. (2002). Invasion of mammalian host cells by
Plasmodium sporozoites. Bioessays: 149─156.
Nurul Aiezzah, Z., Noor, E. & Hasidah, M. S. (2010). Suppression of Plasmodium
berghei parasitemia by LiCl in an animal infection model. Tropical
Biomedicine: 624─631.
Peters, W. (1975). The chemotherapy of rodent malaria, XXII. The value of
drug-resistant strains of P. berghei in screening for blood
schizontocidal activity. Annals Tropical Medicine and Parasitology:
155─171.
Rasoanaivo, P., Deharo, E., Ratsimamanga-Urverg, S. & Frappier, F. (2004).
Traditional Medicinal Plants and Malaria: Guidelines for the Nonclinical
Evaluation of the Efficacy of Tradisional Antimalaria. CRC Press.
Suhaini, S., Liew, S.Z., Norhaniza, J., Lee, P.C., Jualang, G., Embi, N. & Hasidah,
M.S. (2015). Antimalarial and anti-inflammatory effects of Gleichenia
truncata are mediated through inhibition of GSK3β. Tropical
Biomedicine: 1─15
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Towbin, H., Staehelin, T. & Gordon, J. (1979). Electrophoretic transfer of
proteins from polyacrylamide gels to nitrocellulose sheets: procedure
and some applications. Proceedings of National Academy of Science
USA: 4350─4354.
105
PEMILIHAN DAN PENGHASILAN BAKA LIMAU PURUT (CITRUS HYSTRIX)
BERMUTU TINGGI
MA Farah Fazwa, J Mailina, MA Nor Azah, M Nur Nazihah, A Mohd. Zaki, SB
Syafiqah Nabilah, S Norhayati & L Mohd Asri
Tel: 03-6279 7332
ABSTRAK
Pokok limau purut tergolong di dalam keluarga Rutaceae. Walaupun pokok ini
berasal dari benua Asia tetapi pada hari ini ia banyak ditanam di Malaysia dan
Thailand. Minyak pati dari daun limau purut (Citrus hystrix) sering digunakan di
dalam industri pewangi, aromaterapi, perasa, makanan dan perubatan
tradisional. Melihat kepada kepentingan spesies ini di dalam industri herba
negara, satu kajian bagi mendapatkan baka-baka berkualiti dari segi hasil
minyak pati dan kandungan sitronellalnya telah di jalankan oleh Program
Membaikbiak Tumbuhan dari Institut Penyelidikan Perhutanan Malaysia
(FRIM). Sebanyak 220 pokok ibu dari 11 populasi limau purut di Semenanjung
Malaysia telah disaring kuantiti dan kualiti minyak patinya. Secara
keseluruhannya, kertas kerja ini membincangkan mengenai i) kaedah
pengumpulan sampel, ii) pengekstrakan minyak pati secara mobil, iii)
penyaringan kandungan kimia minyak pati, iv) pemilihan baka berkualiti dan
seterusnya iv) kaedah pembiakan baka terpilih dan v) penubuhan bank klonal.
Hasil daripada kajian ini ialah baka-baka limau purut yang berkualiti dapat
diperolehi dan dijadikan sebagai sumber stok tanaman untuk aktiviti
pembiakbakaan terhadap spesies ini pada masa hadapan.
Kata kunci: minyak pati, sitronellal, kuantiti dan kualiti, stok tanaman,
pembiakbakaan
PENGENALAN
Citrus hystrix, lebih dikenali nama tempatannya sebagai limau purut adalah
satu daripada spesies liar di Malaysia. Ia juga dikenali dengan nama “Wild
Lime” dalam bahasa Inggeris, “Kaffir Lime” dalam bahasa Danish dan “Som
Makrut” di Thailand. Tanaman saka ini berasal dari rantau Asia Tenggara dan
telah banyak ditanam di Indonesia, Thailand dan Malaysia. Spesies ini tumbuh
liar di tanah kering dan terdedah kepada cahaya matahari. Pertumbuhan limau
adalah sekitar 3-5 meter dan mencapai sehingga ketinggian 30 meter, tetapi
jika dibandingkan dengan spesies limau lain, pertumbuhannya adalah perlahan.
106
Daun limau purut mempunyai kepanjangan antara 7.5–10.0 cm dan
mempunyai tangkai bersayap yang memberi gambaran dua helai daun yang
dicantumkan hujung ke hujung (Yahaya & Ahmad Puat 2005). Buahnya
berbentuk “pear” dan mencapai kepanjangan sehingga 10 cm dan mempunyai
diameter antara 5.0 cm hingga 7.5 cm. Buahnya berwarna hijau dan bertukar
kepada kekuningan apabila telah masak, berkedut dan berpermukaaan kasar
(Chin & Yong 1980).
Daun limau purut digunakan sebagai perasa dalam masakan Thailand
seperti tomyam. Bahagian daun limau purut juga digunakan sebagai salah satu
bahan untuk mandian wap. Kajian juga menunjukkan daun limau purut sebagai
sumber berkesan penggalak antitumor atau pencegah kanser. Buah limau
purut digunakan secara tradisi untuk mencuci rambut dan air dari buahnya
digunakan untuk membunuh pacat, mencuci mulut dan menguatkan gusi. Kulit
buahnya pula sering digunakan untuk membuat jamu, dan memberikan aroma
terhadap masakan berdaging. Minyak pati limau purut yang diekstrak dari
bahagian daun dan kulit buah mempunyai aroma khusus digunakan untuk
aromaterapi, nutraseutikal dan produk penjagaan diri. Minyak pati ini juga
mempunyai aktiviti antiseptik, astrigen dan antibakteria yang boleh merawat
luka dan lebam (Yahaya et al. 2005). Ia juga baik untuk sakit sendi dan sakit
kepala (Khatijah 2006). Dalam kajian Nor Azah et al. (2004), bahagian buah dan
kulit buah limau purut telah diekstrak minyak patinya dan dirumus dengan
beberapa surfaktan dan bahan aditif untuk menghasilkan produk pembersih
tangan dan produk penjagaan diri.
Melihat kepada kepentingan spesies ini di dalam industri herba negara,
satu kajian bagi mendapatkan baka-baka berkualiti dari segi hasil minyak pati
dan kandungan sitronellalnya telah dijalankan oleh Program Membaikbiak
Tumbuhan, Institut Penyelidikan Perhutanan Malaysia (FRIM) melalui kaedah
penyaringan dan pemilihan. Pemilihan induk atau baka yang baik merupakan
langkah penting dalam mana-mana aktiviti pembiakbaikan tumbuhan sebelum
dibuat perambatan tampang dan sebagainya. Antara tujuan pemilihan juga
adalah untuk memastikan keselamatan diversiti spesies dari terhakis dan juga
untuk memenuhi koleksi kutipan.
Stok baka limau purut ini ditempatkan di FRIM bagi tujuan
pembiakbakaan masa hadapan dan seterusnya untuk tujuan komersialisasi.
Hasil kajian ini juga boleh membantu meningkatkan usaha dalam penghasilan
bahan tanaman yang kini semakin kurang dititikberatkan dalam bidang R&D
negara.
107
KAEDAH KAJIAN
Pengutipan Sampel Kajian
Sebanyak 180 pokok ibu dari 9 populasi limau purut di Semenanjung Malaysia
telah disaring daunnya bagi mendapatkan kuantiti dan kualiti minyak pati.
Populasi tersebut adalah dari 1) Yan, Kedah 2) Balik Pulau, Pulau Pinang 3)
Banting, Selangor; 4) Teluk Intan, Perak; 5) Kuala Pilah, Negeri Sembilan; 6)
Jasin, Melaka; 7) Raub, Pahang 8) Kuala Berang, Terengganu, dan 9) Ketereh,
Kelantan. Setiap populasi dinamakan mengikut kod-kod yang berbeza. Pokokpokok ibu yang dipilih mempunyai sifat-sifat fenotip yang baik seperti jumlah
daun dan buah yang lebat serta bebas dari serangan penyakit dan perosak
(Rajah 1). Bagi mengelakkan berlakunya pengurangan pada hasil minyak pati
dalam daun yang mungkin berlaku semasa dalam perjalanan, semua daun
dikutip dan disaring secara langsung di lapangan atau dinamakan “mobile lab”.
Rajah 1. Contoh pokok ibu yang dipilih untuk tujuan pemilihan baka berkualiti
terletak di Yan, Kedah.
Penyulingan Minyak Pati Sampel Kajian
Penyulingan minyak pati dijalankan sebaik sahaja sampel daun dikutip dari
lapangan. Daun matang yang sesuai diperolehi dengan memotong daun-daun
tersebut dari ranting pokok. Proses penyulingan bagi setiap sampel dijalankan
selama 6 jam menggunakan peralatan jenis Clavenger. Sebanyak 3 replikasi
digunakan bagi setiap sampel pokok ibu (Rajah 2).
108
Rajah 2. Gambar menunjukkan proses penyulingan minyak pati yang
dijalankan secara “mobile lab” di setiap lokasi kajian.
Penentuan Kandungan Kimia Minyak Pati
Analisa kualitatif kromatografi gas (GC) dan kromatografi gas jisim spektrometri
(GCMS) dijalankan menggunakan alat Shimadzu GC2010 Plus dan Agilent
Technologies 7890A/5975C MSD (Rajah 3) dengan masing-masing
menggunakan kolum kapilari BP-5 (30 m x 0.25 mm, 0.25 mm tebal filem) dan
HP-5MS (30 m x 0.25 mm, 0.25 mm tebal filem). Kromatografi gas dilengkapi
dengan FID dan menggunakan suntikan split mode. Helium digunakan sebagai
gas pembawa dengan kadar tetapnya 1ml/min. Suhu pengesan ditetapkan
pada 250 °C. Peralatan GC diprogramkan pada awalnya dengan suhu 60°C
selama 10 minit dan kemudian ditingkatkan ke suhu 230 °C selama 1 minit
dengan kadar aliran 3°C/min. Puncak komponen dan masa penahanan diukur
menggunakan integrasi elektronik. Bagi GC/MS, program suhu yang sama
ditetapkan seperti pada peralatan GC. Pengenalpastian komponen-komponen
kimia adalah dengan membandingkan nilai kovat indek komponen di dalam
sampel dengan nilai rujukan (Adams 2005) dan juga perbandingan spektrum
jisim sampel dengan spektrum di pangkalan data.
Jumlah Minyak Pati
Keputusan hasil minyak pati yang diperolehi dari 9 populasi adalah seperti di
dalam Jadual 1. Hasil menunjukkan kesemua pokok ibu yang disaring
mempunyai julat di antara 0.9 hingga 4.7%. Jadual 2 menunjukkan purata
minyak pati yang dihasilkan dari 9 populasi di mana populasi Kuala Berang
mencatatkan jumlah minyak pati tertinggi (4.2%).
109
Jadual 1. Julat hasil minyak pati dari yang disaring dari 9 populasi kajian
Populasi
Julat minyak pati (%)
Yan
1.70-4.6
Balik Pulau
1.25-2.2
Teluk Intan
1.30-2.7
Banting
3.10-4.5
Kuala Pilah
1.30-4.7
Jasin
0.90-2.2
Raub
2.20-4.2
Kuala Berang
1.40-2.7
Ketereh
2.20-3.6
Jadual 2. Purata jumlah minyak pati limau purut dari 9 populasi kajian
Populasi
Purata jumlah minyak pati (%)
Yan
3.2%
Balik Pulau
3.4%
Teluk Intan
3.7%
Banting
3.6%
Kuala Pilah
3.2%
Jasin
2.8%
Kuala Berang
4.2%
Raub
3.2%
Ketereh
3.0%
Jumlah kompaun sitronellal dan lain-lain kompaun utama
Analisa GC dan GC/MS telah mencatatkan sitronellal sebagai sebatian utama
yang hadir di dalam minyak pati limau purut. Didapati populasi Kuala Berang
juga memberikan purata tertinggi (83.7%) berbanding lain-lain populasi.
Terdapat juga tiga jenis sebatian kimia utama yang lain dalam kesemua minyak
pati limau purut selain daripada sitronellal iaitu sitronellol, linalool dan
sitronellil asetat (Jadual 3). Sitronellal (C10H18O) adalah sebatian yang digunakan
sebagai kriteria dalam pemilihan baka bermutu tinggi. Sitronellal atau sinonim
dengan formula molekul 3,7-dimethyl-6-octanal merupakan sebatian
monoterpena yang berfungsi memberikan bauan limau di dalam minyak pati
limau (Takeshi et al. 2004).
110
Jadual 3. Kandungan kimia utama (%) yang dijumpai dalam minyak pati limau
purut yang disaring dari 9 populasi di Semenanjung Malaysia
Populasi
Sebatian
Sitronellal
Sitronellol
Linalool
Sitronellil
asetat
Yan
Balik
Pulau
Teluk
Intan
Banting
Kuala
Pilah
Jasin
Kuala
Berang
Raub
Ketereh
68.1
10.0
2.2
74.4
4.0
2.2
82.0
5.8
2.5
72.7
9.6
2.5
54.4
10.7
1.5
76.9
8.6
2.2
83.7
4.8
2.7
63.6
9.4
1.9
77.5
9.0
2.3
4.2
3.4
2.9
3.7
3.8
3.7
2.1
2.7
2.9
Berdasarkan keseluruhan keputusan yang diperolehi, didapati sebanyak 10
pokok ibu telah dipilih sebagai baka yang berkualiti dan bermutu kerana
berupaya menghasilkan jumlah minyak pati dan kandungan sitronellal yang
tinggi. Baka-baka terpilih ini akan dijadikan sebagai sumber stok bahan
tanaman pada masa hadapan. Semua baka-baka terpilih ini telah dibiakkan
secara tut bagi mendapatkan anak-anak klon dan kini ditempatkan di FRIM
untuk tujuan pembiakbakaan dan komersialisasi.
Rajah 3. Klon bank limau purut yang ditubuhkan di FRIM bagi menempatkan
baka-baka limau purut yang berkualiti
KESIMPULAN
Bagi pembiakbaka yang ingin menjalankan program biakbaka, sepuluh pokok
terpilih ini sangat berguna untuk diperbanyakkan melalui kaedah perambatan
tampang. Hasil anak-anak klon boleh ditanam untuk penubuhan bank klonal
atau penubuhan germplasma. Kelak dengan adanya aktiviti sebegini, baka-baka
terbaik yang menghasilkan sitronellal iaitu kompaun terbaik dalam industri
pewangi dapat dikekalkan. Dengan itu, sumber minyak pati untuk kegunaan
111
industri ini akan dapat diperolehi dengan mudah dan produk yang dikeluarkan
adalah dari bahan yang berkualiti tinggi.
RUJUKAN
Adam, R.P. (2005). Identification of Essential oil Components by Gas
Chromatography/Mass spectrometry, 4th Edition. Allured Business
Media: Carol Stream, Illinois.
Chin, H.F. & Yong, H.S. (1980). Malaysian Fruits in Color. Tropical Press Sdn.
Bhd, Kuala Lumpur, Malaysia. Pp 52.
Khatijah, H. (2006). Anatomical atlas of Malaysian Medicinal Plants Volume 1.
Universiti Kebangsaan Malaysia, Bangi. Pp : 58-61.
Nor Azah, M.A., Mastura, M., Zaridah, M.Z., Mailina, J., Mohamad Shahidan.,
Norseha, A., Zainon, A.S., Norhanan, M.Y., Abdul Majid, J. & Abu Said, A.
(2004). Potential application of Citrus hystrix essential oils for personal
care products. Prosiding Hasil Kajian IRPA RMK8 2004 FRIM. Pp :
273─277.
Takeshi, Y., Akemi, S., Tatsuya, O., Hiroyuki, M., Miharu, O. & Tsuneyoshi, K.
(2004). Flavour and Fragrance Journal Volume 19, Issue 2. Pp : 121─133.
Yahaya H. & Ahmad Puat, N. (2005). Limau purut. Penanaman Tumbuhan
Ubatan dan Beraoroma. Cetakan pertama. Eds. Musa Yaacob, Muhamad
Ghawas Maarif & Mansor Puteh. Institut Penyelidikan dan Kemajuan
Pertanian Malaysia. Pp 109.
112
PENGELUARAN DAN PENILAIAN MINYAK DAUN KAYU MANIS
MALAYSIA
H Norma1, N Noor Ismawaty1, M Erny Sabrina2, AR Zuraida3, EA Engku
Hasmah 4, M Razali2, WM Wan Zaki1, MA Mohd Shukri5, B Ibrahim1, T
Sulaiman1, AR Zulkifli1 & M Osman1
1
Pusat Hortikultur, Institut Penyelidikan dan Kemajuan Pertanian Malaysia
2
Pusat BE, Institut Penyelidikan dan Kemajuan Pertanian Malaysia
3
Pusat BT, Institut Penyelidikan dan Kemajuan Pertanian Malaysia
4
Pusat RI, Institut Penyelidikan dan Kemajuan Pertanian Malaysia
5
Pusat BB, Institut Penyelidikan dan Kemajuan Pertanian Malaysia
Ibu Pejabat MARDI , 43400 Serdang, Selangor
ABSTRAK
Kandungan kimia minyak dedaun kayu manis biasanya adalah eugenol dan
sinnamik aldehid. Bagi penyulingan secara stim skala industri, bahan campuran
dedaun dan cabang ranting kayu manis memberikan 0.4–0.5% hasil minyak
pati, manakala skala makmal penyulingan secara hidro dedaun kayu manis
sahaja memberikan hasil penyulingan sebanyak 1.6%. Minyak dedaun kayu
manis bewarna kekuningan cerah dan berkilau. Komponen utama dalam
minyak pati daun kayu manis adalah kumpulan eugenol dan sinnamik aldehid.
Aroma minyak pati daun kayu manis adalah sangat segar, manis serta sedikit
tajam. Bahan ini merupakan bahan perisa dan wangian (Flavour & Fragrance
(F&F)) bagi ramuan makanan, minuman, minyak wangi dan kosmetik.
Kata kunci: Minyak pati, GC-MS-ToF, eugenol, sinnamik aldehid, penyulingan,
daun kayu manis, perisa dan wangian
PENGENALAN
Pokok kayu manis sangat banyak kegunaannya dalam industri makanan,
minuman, konfeksioneri, kosmetik, perisa, wangian, antibakteria, penghalau
dan pencegah serangga. Secara amnya bahan mentah ini dikategorikan dalam
kumpulan Flavour and Fragrance (F&F) dan juga biopestisid. Bahan utama yang
boleh diperolehi daripada kulit pokok atau dedaun pokok kayu manis adalah
minyak pati (essential oil). Kajian ini memfokuskan kepada peggunaan dedaun
daripada pokok kayu manis Cinnamomun zeylanicum di MARDI Kuala Linggi,
Melaka bagi penghasilan minyak pati. Alatan penyulingan skala industri
digunakan bagi pengeluaran secara pukal manakala skala makmal dikhususkan
113
bagi penilaian penyelidikan dan pembangunan (R&D) minyak pati dedaun kayu
manis.
Beberapa spesies kayu manis atau Cassia adalah seperti Cinnamomum
cassia (cassia atau Chinese cinnamon) merupakan spesies yang paling banyak
ditemui; C. burmannii (Korintje, Padang cassia, atau Indonesian cinnamon); C.
loureiroi (Saigon cinnamon, Vietnamese cassia, atau Vietnamese cinnamon)
dan C. verum (Sri Lanka cinnamon atau Ceylon cinnamon) (Culinary Herbs and
Spices 2015). Kayu manis banyak digunakan dalam pembuatan bahan perisa
makanan dan juga aromaterapi serta minyak wangi. Kulit batang dan dedaun
kayu manis dijadikan rempah-ratus dan perisa makanan dan minuman. Dedaun
kayu manis mengandungi kandungan eugenol yang tinggi sehingga 86%
(Amarasinghe et al. 2011), berbanding kulit batang kayu manis yang tinggi
dengan sinnamaldehid 45.13% (El-Baroty et al. 2010), 72.0% (Amarasinghe et
al. 2011), 63.1% (Inouye 2001).
Kajian ini memberikan penerangan berkaitan dengan profil kimia daun
kayu manis C. zeylanicum bagi membangunkan produk-produk berasaskan
kayu manis di Malaysia.
BAHAN DAN KAEDAH
Bahan mentah dedaun kayu manis dituai di MARDI Kuala Linggi dan disuling
menggunakan penyulingan stim berskala 100 kg. Kandungan bahan mentah
yang disuling secara industri adalah merangkumi dahan, ranting dan dedaun.
Manakala bagi hasil sebenar kandungan minyak dedaun adalah secara
penyulingan berskala makmal dengan menggunakan apparatus Clavenger (2 L).
Analisis GC-MS-Tof (1D) digunakan untuk pengenalpastian kandungan kimia
beberapa aroma kimia makanan dan bahan wangian. Pengesan ToF ini adalah
singkatan bagi time-of-flight mass spectrometer menggunakan alat GC-MS
Pegasus 4D, Leco Corporation, St. Joseph, MI, USA. Parameter penggunaan
kolum bagi GC-MS-Tof adalah 1D column model ZB-5MS, panjang 30 m,
diameter 0.25 mm, ketebalan filem dalaman 0.25 μm, fasa utama column 5%
Polysilarylene dan 95% Polydimethylsiloxane (Zebron, Phenomenex
Corporation).
114
Kandungan minyak pati daun kayu manis yang diperolehi secara skala makmal
adalah sehingga 1.6%, manakala bahan campuran dedaun, dahan dan ranting
memberikan hasil minyak yang rendah dengan hanya 0.4–0.5%. Namun
operasi penyulingan berskala besar boleh menghasilkan sehingga 500 ml
minyak daun kayu manis premium bagi penyulingan 1 vessel/batch. Rajah 1
menunjukkan produk minyak pati kayu manis bergred premium iaitu campuran
dahan dan ranting bagi pengeluaran produk secara pukal menggunakan
penyulingan skala kilang di MARDI.
Rajah 1. Satu liter minyak pati kayu manis Malaysia gred premium daripada
campuran dahan dan ranting)
Analisis GC-MS-ToF menunjukkan kandungan bahan kimia aromatik yang
tinggi bagi kumpulan eugenol (Rajah 2) dan juga benzyl benzoate. Eugenol
boleh bertukar menjadi sebatian iso-eugenol dan memberikan perisa makanan
lebih baik.
Rajah 2. Komposisi kimia utama (eugenol dan iso-eugenol) dalam minyak daun
kayu manis
115
Daripada analisis ini kandungan kumpulan eugenol dan benzyl benzoate
masing-masing adalah sebanyak 56.0% dan 14.2% (Rajah 3). Di samping itu
komponen kimia terpilih dan penting dalam industri minyak wangi juga telah
dikenal pasti iaitu eugenol asetat (6.5%), E-sinnamaldehid (sinnamil asetat
(2.4%) dan hidrosinnamil asetat (0.2%).
Bahan aroma sinnamaldehid ini juga berfungsi dan efektif sebagai bahan
anti-mikrob menurut laporan kajian serapan gas terhadap Escherichia coli,
Staphylococcus aureus, Streptococcus pyogenes, S. pneumoniae dan
Haemohilus influenza (Inouye 2001). El-Baroty et al. (2010) pula melaporkan
penggunaan minyak kayu manis di antara 20–100 µg/ml boleh bertindak
sebagai bahan pengawet asli dan menghalang pengoksidaan bahan (lipid
peroxidation). Secara tidak langsung eugenol dan sinnamik aldehid yang
terkandung dalam minyak dedaun dan ranting kayu manis ini berpotensi
sebagai agen anti-oksida dan anti-bakteria.
Rajah 3. Profil 1D GC-MS Tof bagi minyak daun kayu manis Malaysia
KESIMPULAN
Minyak pati daun kayu manis yang dikeluarkan di Malaysia mempunyai potensi
sebagai bahan perisa kerana kandungan kumpulan aktif eugenol yang tinggi
dan bahan sinnamaldehid. Produk berasaskan minyak pati boleh menggantikan
produk oleoresin kayu manis yang banyak menggunakan pelarut dan
mengandungi residu pelarut yang mungkin boleh menjejaskan kesihatan
116
pengguna. Kajian ini melengkapkan lagi data minyak pati daun kayu manis yang
berpotensi bagi industri minyak pati di Malaysia, serta boleh memberikan data
sokongan kepada cadangan Teknologi Penanaman dan Pengeluaran Bahan
herba F&F dalam Rancangan Malaysia kesebelas (RMK11), sekiranya tumbuhan
beraroma terpilih sebagai herba baharu dalam EPP1.
PENGHARGAAN
Penulis mengucapkan ribuan terimakasih kepada pihak pengurusan dan
kakitangan R&D Pusat HR, Pusat BE dan Pusat BT MARDI HQ, kakitangan
MARDI Kuala Linggi dan Makmal CRIM, UKM dan yang terlibat secara langsung
dan tidak langsung bagi bantuan dan menjayakan artikel ini.
RUJUKAN
Amarasinghe, L.D., Wijesinghe, W.K.A.G.A. & Jayawardhane, B. K. (2011).
Efficacy of Essential Oils from Bark and Leaf of Cinnamomum
Zeylanicum on Root Knot Nematode, Meloidogyne Graminicola in Rice
Seedlings and Young Rice Plants. J sci.univ.kelaniya 6 : 45─54.
Culinary Herbs and Spices, the Seasoning and Spice Association. 2015.
http://www.seasoningandspice.org.uk/ssa/background_culinary-herbsspices.aspx. [23 September 2015].
El-Baroty, G.S., Abd El-Bakri H.H., Farag, R.S. & Saleh, M.A. (2010).
Characterization of Antioxidant and Antimicrobial Compounds of
Cinnamon and Ginger Essential Oils. African J. of Biochem. Research
Vol. 4(6): 167─174.
Inouye, S, Takizawa, T. & Yamaguchi, H. (2001). Antibacterial Activity of
Essential Oils and Their Major Constituents Against Respiratory Tract
Pathogens by Gaseous Contact. J. of Antimicrobial Chemotherap 47:
565─573.
117
PEMBIAKAN AKSESI KACIP FATIMAH (LABISIA PUMILA VAR. ALATA)
TERPILIH MELALUI KAEDAH KERATAN DAN PENILAIAN PENGELUARAN
PUCUK PADA PERINGKAT TAPAK SEMAIAN
M Nur Nazihah, MA Farah Fazwa, S Norhayati, SB Syafiqah Nabilah, L Mohd
Asri & Z Mohd Zaini
Institut Penyelidikan Perhutanan Malaysia (FRIM), 52109, Kepong, Selangor.
Tel: 03-6279 7332
ABSTRAK
Sejumlah tiga belas aksesi kacip fatimah (Labisia pumila var. alata) yang
mempunyai kandungan fenolik yang tinggi telah dibiakkan melalui kaedah
keratan daun. Tujuan kajian ini dijalankan adalah untuk menilai keupayaan
pengakaran dan menganalisa prestasi pertumbuhan aksesi kacip fatimah yang
dihasilkan daripada kaedah keratan daun. Melalui kaedah ini, daun kacip
fatimah dirawat menggunakan hormon pengakaran (Seradix 1) dan dibiakkan
di dalam rumah keratan dalam sistem renjisan selama 17 minggu. Setelah
dirawat dengan hormon, keratan yang berakar telah dipindahkan ke dalam
polibeg yang mengandungi media tanaman campuran tanah: kompos: sabut:
pasir (3: 1: 1: 1) dan dipindahkan ke tapak semaian dengan 70% teduhan. Data
seperti kadar kelangsungan hidup, jumlah keratan berakar dan jumlah pucuk
yang terhasil diperhatikan sehingga minggu ke 30. Daripada keputusan yang
diperolehi, aksesi LP7 mencatatkan peratus pengakaran tertinggi (91%) diikuti
oleh LP15 dan LP28 (87%), LP17 (85%) dan LP12 (84%). Manakala, LP15, LP17
dan LP28 menghasilkan bilangan keratan berakar tertinggi dengan purata
masing-masing 13.00 ± 1.00, 13.00 ± 0.58 dan 13.00 ± 1.00. Dari segi
pengeluaran pucuk, LP15 dan LP21 masing-masing menghasilkan bilangan
pucuk yang tinggi iaitu 2.43 ± 0.271 dan 2.43 ± 0.386. Hasil kajian ini dapat
membantu para pembiakbaka dan pengusaha herba mengenal pasti aksesiaksesi yang terbaik dari segi pertumbuhan bermula dari peringkat awal
pertumbuhan (peringkat nurseri) sebelum ianya ditanam di lapangan.
Pemilihan baka tanaman yang baik dapat menjamin pengeluaran stok bahan
tanaman secara berterusan.
Kata kunci: rumah keratan sistem renjisan, media tanaman, kadar
kelangsungan hidup, bilangan keratan berakar, pengeluaran pucuk, pemilihan
baka
118
PENGENALAN
Malaysia terkenal dengan pelbagai jenis herba yang selalu digunakan dalam
perubatan tradisional. Labisia pumila atau lebih dikenali sebagai kacip fatimah
merupakan salah satu herba tradisi yang terkenal dalam kalangan wanita
Melayu terutamanya bagi wanita mengandung. Air rebusan kacip fatimah
mempunyai banyak khasiat terutamanya dapat mengurangkan kesakitan
semasa bersalin dan mempercepat proses bersalin (Ibrahim & Hawa 2011). Air
kacip fatimah juga dapat memberi tenaga kepada ibu hamil sepanjang proses
bersalin (Wan Ezumi et al. 2007). Selain itu, kacip fatimah juga dikatakan dapat
menyembuhkan penyakit seperti disentri, senggugut, kembung perut dan
gonorea.
Kacip fatimah dilaporkan mempunyai kadar pertumbuhan yang
perlahan di dalam habitat semula jadinya (Mohd Noh et al. 2002). Penyebaran
melalui biji benih secara kaedah konvensional mengambil masa yang lama
untuk berlakunya percambahan. Tambahan pula, bahan mentah bagi spesies
ini terus dieksploitasi untuk pelbagai tujuan. Dalam jangka masa yang panjang,
bahan tanaman ini mungkin akan berkurangan di dalam habitat semula jadi jika
sedikit usaha dilakukan untuk mengatasi masalah ini.
Kajian ini dijalankan bertujuan untuk menghasilkan sejumlah besar
anak pokok melalui kaedah keratan daripada beberapa aksesi kacip fatimah
terpilih. Beberapa kajian telah dilakukan oleh Aminah et al. (2008), Rozihawati
et al. (2005) dan Farah Fazwa et al. (2013) ke atas spesies ini menggunakan
kaedah keratan dan hasil kajian tersebut menunjukkan respon yang baik
terhadap pertumbuhan akar dan pucuk. Diharapkan hasil kajian ini akan dapat
digunakan sebagai rujukan bagi membantu pembiakbaka dan pengusaha herba
dalam menghasilkan stok bahan tanaman yang berterusan untuk masa
hadapan.
BAHAN DAN KAEDAH
Bahan Tanaman
Sebanyak 13 aksesi kacip fatimah telah diambil dari tiga lokasi hutan simpan
yang berbeza di Semenanjung Malaysia iaitu HS Tembat, Kuala Berang,
Terengganu (LP5, LP6, LP11, LP21); HS Bukit Larut, Taiping, Perak (LP7, LP9,
LP12, LP10, LP13, LP15, LP30); dan HS Batu Papan, Gua Musang, Kelantan
(LP17, LP28). Kesemua pokok ibu ini ditanam sebagai koleksi germplasma yang
telah ditubuhkan di FRIM. Pokok-pokok ibu tersebut dipilih berdasarkan jumlah
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daun iaitu lebih daripada lima dan juga daun yang sihat tanpa sebarang
penyakit.
Pembiakan Kacip Fatimah Melalui Teknik Keratan Daun
Daun diambil dari pokok-pokok ibu terpilih yang ada di dalam koleksi
germplasma kacip fatimah. Daun dipotong mengikut saiz 30 cm2. Hormon
pengakaran (Seradix 1) telah diletakkan pada pangkal keratan. Keratan
dibiakkan di dalam rumah keratan sistem renjisan selama 17 minggu. Pasir
sungai steril digunakan sebagai media tanaman. Sebanyak 585 keratan telah
dihasilkan daripada 13 aksesi. Keratan tersebut ditanam mengikut kaedah
Rekabentuk Rawak Blok Lengkap (RCBD) dengan tiga replikasi (Rajah 1).
Daun dipotong
2
saiz 30 cm .
mengikut
Hormon
diletakkan
keratin.
Seradix
1
pada pangkal
Keratan dibiakkan di dalam
rumah
keratan
sistem
renjisan.
Rajah 1. Kaedah keratan daun
Penanaman pada Peringkat Tapak Semaian
Selepas dibiakkan selama 17 minggu di rumah keratan, keratan yang berakar
dipindahkan ke dalam polibeg bersaiz 8’ x 6’ dan diletakkan di tapak semaian.
Media penanaman yang digunakan ialah campuran tanah: kompos: sabut: pasir
(3: 1: 1: 1). Beberapa data persekitaran di tapak semaian seperti suhu,
kelembapan relatif dan keamatan cahaya direkodkan dari jam 8.30 pagi
sehingga 4.00 petang. Tapak semaian dilitupi jaring hitam dengan teduhan
sebanyak 70%.
120
Pengumpulan Data
Penilaian pertumbuhan akar
Data prestasi pertumbuhan akar telah dicatat pada minggu ke 6 hingga minggu
17. Data yang dikumpul tertakluk kepada analisis varians (ANOVA) untuk
menentukan kesan aksesi terhadap jumlah pertumbuhan akar dalam 17
minggu menggunakan perisian SPSS versi 16.0.
Penilaian pertumbuhan pucuk
Data prestasi pertumbuhan pucuk telah dicatat pada minggu 22 hingga minggu
30. Data telah direkod setiap dua bulan sehingga minggu 30. Parameter yang
diperhatikan ialah bilangan daun. Data telah dianalisa menggunakan perisian
SPSS versi 16.0 untuk menilai dan membandingkan prestasi pertumbuhan
antara aksesi.
Jadual 1 menunjukkan purata bilangan akar, panjang akar dan peratus
pengakaran (%) bagi tiga belas aksesi kacip fatimah. Analisis varian (ANOVA)
terhadap bilangan akar menunjukkan tiada perbezaan yang signifikan antara
kesemua tiga belas aksesi. Purata bilangan akar bagi semua aksesi adalah
antara 2.56 hingga 4.78. Manakala analisis varian terhadap panjang akar juga
menunjukkan tiada perbezaan yang signifikan antara aksesi. Purata panjang
akar bagi kesemua aksesi adalah antara 3.06 hingga 4.91. Analisa peratus
pengakaran menunjukkan terdapat perbezaan yang signifikan antara LP7, LP11
dan LP5.
121
Jadual 1. Data pertumbuhan akar tiga belas aksesi kacip fatimah sehingga
minggu ke 17
Aksesi
Bilangan akar
Panjang akar (cm)
Peratus pengakaran (%)
LP5
2.78ab ± 0.364
3.06a ± 0.584
27.0d
LP6
2.56b ± 0.412
4.64a ± 0.708
73.0abc
LP7
4.11ab ± 0.992
4.31a ± 0.972
91.0a
LP9
4.33ab ± 0.553
4.77a ± 0.424
78.0abc
LP10
3.67ab ± 0.373
4.00a ± 0.367
71.0abc
LP11
3.78ab ± 0.465
3.80a ± 0.439
56.0c
LP12
3.44ab ± 0.648
4.20a ± 0.745
84.0ab
LP13
3.78ab ± 0.703
4.04a ± 0.701
71.0abc
LP15
4.22ab ± 0.465
4.68a ± 0.659
87.0ab
LP17
4.78ab ± 0.572
4.40a ± 0.283
85.0ab
LP21
3.89ab ± 0.389
4.91a ± 0.312
78.0abc
LP28
3.67ab ± 0.471
4.60a ± 0.336
87.0ab
LP30
4.56ab ± 1.132
3.86a ± 0.870
60.0bc
Jadual 2 menunjukkan data pertumbuhan daun bagi tiga belas aksesi
kacip fatimah sehingga minggu 30. Berdasarkan ujian Duncan yang diperoleh
menunjukkan terdapat perbezaan yang signifikan pada bilangan pucuk bagi
kesemua aksesi. LP15 dan LP21 masing-masing mencatatkan penghasilan
pucuk tertinggi. Manakala LP17 menunjukkan pertumbuhan pucuk yang paling
rendah. Secara umumnya, pertumbuhan akar dan pucuk yang lambat bagi
beberapa aksesi mungkin disebabkan oleh bekalan nutrien yang terhad
(Aminah et al. 2008). Terdapat juga beberapa keratan yang didapati telah mati
mungkin disebabkan oleh serangan kulat atau kandungan karbohidrat yang
rendah di bahagian keratan (Rozihawati et al. 2005). Pokok kacip fatimah juga
berkemungkinan sensitif pada cahaya matahari yang boleh menyebabkan daun
tersebut layu dan kering. Data persekitaran di tapak semaian seperti suhu dan
kelembapan relatif yang direkodkan dari jam 8.30 pagi hingga 4.00 petang
adalah seperti Jadual 3.
122
Jadual 2. Data pertumbuhan daun tiga belas aksesi kacip fatimah sehingga
minggu 30
Aksesi
Bilangan pucuk
LP5
1.16b ± 0.254
LP6
1.13b ± 0.152
LP7
1.50b ± 0.184
LP9
2.41a ± 0.281
LP10
1.55b ± 0.152
LP11
1.54b ± 0.346
LP12
1.39b ± 0.189
LP13
1.90ab ± 0.269
LP15
2.43a ± 0.271
LP17
1.07b ± 0.219
LP21
2.43a ± 0.386
LP28
1.46b ±0.187
LP30
1.84ab ± 0.465
*Purata diikuti oleh huruf yang sama tidak mempunyai perbezaan yang signifikan
p<0.05
Jadual 3. Data persekitaran di tapak semaian dari jam 8.30 pagi hingga 4.00
petang
Suhu (°C)
26 ─ 28
Kelembapan relatif (%)
56 ─ 68
KESIMPULAN
Hasil penyelidikan menunjukkan bahawa teknik keratan sangat sesuai
digunakan dalam pembiakan 13 aksesi kacip fatimah terpilih yang digunakan
dalam kajian ini. Pengenalpastian baka-baka yang mudah dibiakkan ini adalah
penting untuk tujuan penghasilan secara komersial di masa hadapan. Ini
bermakna baka bukan sahaja perlu berkualiti dari segi kandungan kimianya
sahaja tetapi juga baik dari segi pembiakan dan pertumbuhannya.
PENGHARGAAN
Penghargaan terima kasih kepada Kementerian Pertanian dan Industri Asas
Tani Malaysia di atas pembiayaan dana melalui Skim Geran Penyelidikan
Kebangsaan (NH1113A020). Ucapan terima kasih juga ditujukan kepada
kakitangan Program Membaikbiak Tumbuhan, FRIM di atas kerjasama yang
diberi dalam memastikan kelancaran projek ini.
123
RUJUKAN
Aminah, H., Naimah, C.L., Mohd Zaki, A., & Lokmal, N. (2008). Rooted Leaf
Cuttings of Labisia pumila. J. Tropical Med. Plants, 9(1): 593─599.
Farah Fazwa, M.A., Norhayati, S., Syafiqah Nabilah, S.B. & Mohd. Adi Faiz, A.F.
(2013). Evaluation of Rooting Ability of Five Superior Genotypes of
Labisia pumila var. alata on sand media. Proceedings of Soils Science
Conference of Malaysia 2013, 329─333.
Ibrahim, M.H. & Hawa Ze, J. (2011). Photosynthetic Capacity, Photochemical
Efficiency and Chlorophyll Content of Three Varieties of Labisia Pumila
Benth Exposed to Open Field and Greenhouse Growing Conditions,
Acta Physiol. Plant, 33: 2179─85.
Mohd Noh, J., Rezuwan, K., & Md Akhir, H. (2006). Performance of Kacip
Fatimah (Labisia pumila) Production Under Shade House. ISHS Acta
Horticulturae 710: International Symposium on Greenhouse,
Environmental Controls and In-house Mechanization for Crop
Production in the Tropics and Sub-tropics: 399─403.
Rozihawati, Z., Azmy, M. & Aminah, H. (2005). Vegetative Propagation of
Labisia Pumila Through Stem Cuttings. Proceedings of the Seminar on
Medicinal and Aromatic Plants. pp 368─373.
Wan Ezumi, M.F., Amrah, S.S., Suhaimi, A. & Mohsin, S. (2007). Evaluation of
The Female Reproductive Toxicity 0f Aqueous Extract of Labisia pumila
var. alata in Rats. Indian J Pharmacol 2007; 39:30─2.
124
SERANGAN ATTEVA SCIODOXA (ULAT HARIMAU) PADA TANAMAN
TONGKAT ALI DI HUTAN DAN LADANG SEMENANJUNG MALAYSIA
WA Wan Muhd Azrul 1, A Mohd Farid1, AS Sajap.2, S Tosiah3 & M Patahayah1
1
Institut Penyelidikan Perhutanan Malaysia (FRIM), 52109, Selangor, Malaysia
2
Universiti Putra Malaysia, 43000 UPM, Serdang, Selangor, Malaysia
3
Malaysian Agricultural Research and Development Institute, 43400 Serdang,
Selangor, Malaysia
ABSTRAK
Pemantauan kesihatan tongkat ali telah menemukan Atteva sciodoxa (Ulat
harimau) adalah musuh utama tanaman ini. Walau bagaimanapun status
serangan perosak ini pada tanaman tersebut masih belum diketahui secara
kuantitatif. Tujuan pemantauan ini dijalankan adalah untuk menilai kehadiran
dan tahap serangan ulat harimau ke atas pokok tongkat ali di hutan dan di
ladang. Sebanyak 28 petak kajian tongkat ali di Semenanjung Malaysia telah
dipilih dalam program ini yang terdiri daripada 13 kawasan hutan, 9 ladang
selingan dan 6 ladang monokultur. Sejumlah 30 pokok bagi setiap petak kajian
dipilih secara rawak dan dinilai. Kehadiran ulat harimau pada setiap pokok
direkodkan. Peratus kehadiran perosak tersebut dikira berdasarkan bilangan
pokok diserang perosak tersebut didalam petak kajian dan tahap keseriusan
serangan perosak di nilai melalui pendekatan “Damage Severity Index” (DSI).
Lima kategori kerosakan ditetapkan iaitu tahap 0: tiada kerosakan, 1: <25%
kerosakan daun, 2: 25%-50% kerosakan daun, 3: 50% - 75% kerosakan daun
dan 4: >75% kerosakan daun. Hasil pemantauan mendapati kebanyakan ladang
telah ditinggalkan dan tidak diuruskan dengan baik. Kehadiran ulat harimau
pada tanaman tongkat ali di dapati berlaku di semua petak kajian terpilih sama
ada di hutan atau ladang dengan kadar keseriusan yang berbeza. Purata
kehadiran ulat harimau pada tanaman tongkat ali di kawasan hutan adalah
sebanyak 16.67% dengan nilai terendah adalah 3.33% dan tertinggi adalah
83.33% . Di ladang selingan pula, purata kehadiran ulat harimau pada tanaman
tersebut adalah 25.92% (Min. 3.33% dan mak. 80.00%). Manakala, purata
kehadiran perosak tersebut di ladang tongkat ali monokultur adalah 50.56%
(Min.16.67% dan mak. 86.7%) . Keputusan DSI pula menunjukkan, tahap
keseriusan serangan ulat harimau pada tanaman tongkat ali di kawasan hutan
adalah 32.69%, di ladang selingan adalah 33.33% dan di ladang monokultur
adalah 58.33% kerosakan. Pemantauan ini menunjukkan serangan ulat
harimau boleh berlaku pada tongkat ali sama ada di kawasan hutan atau
125
ladang. Di ladang, penjagaan dan pengurusan yang baik dapat membantu
dalam mengawal serangan ulat harimau ini.
Kata kunci: ulat harimau, pemantauan, monokultur, selingan, tanaman
PENDAHULUAN
Eurycoma longifolia Jack, atau nama tempatannya tongkat ali merupakan salah
satu tumbuhan herba yang terkenal di Malaysia. Tumbuhan ini mempunyai
nilai perubatan dalam meningkatkan tahap kesihatan manusia. Permintaan
bahan mentah tongkat ali sangat menggalakkan namun kebanyakannya diambil
daripada hutan asli. Bagi memenuhi permintaan ini, penanaman pokok tongkat
ali secara ladang telah diperkenalkan. Teknik penanaman yang sering
diamalkan oleh para pengusaha adalah tanaman secara monokultur dan juga
secara selingan dengan pokok-pokok pertanian. Penanaman secara begini
boleh mengelakkan kepupusan pokok tongkat ali di dalam hutan akibat
penuaian yang tidak terkawal. Walau bagaimanapun serangan penyakit dan
perosak telah memberi masalah dalam usaha ini. Patahayah et al. (2011) dan
Mohd. Noh (2004) melaporkan beberapa jenis penyakit dan perosak berbahaya
kerap ditemui di ladang tongkat ali. Antaranya adalah sindrom mati mengejut
atau Sudden Death Syndrom (SDS), serangan serangga Atteva sciodoxa (Ulat
Harimau), serangga teritip dan ulat penebuk batang (Zeuzera sp.). Serangan
ulat harimau merupakan perosak utama pokok tongkat ali (Mohd. Noh et al.
2004; Mohd Farid et al. 2014). Perosak ini diperhatikan hadir sepanjang masa
dan merupakan pemakan daun yang agresif. Serangan yang serius mampu
menyebabkan kematian atau pokok menjadi renggeh. Namun masih belum ada
kajian secara menyeluruh dilakukan bagi mengetahui tahap kerosakan akibat
serangan ulat harimau ini di Semenanjung Malaysia. Satu kajian telah
dijalankan bagi menilai status kehadiran dan tahap keterukan serangan ulat
harimau pada tongkat ali di ladang dan di hutan.
BAHAN DAN KAEDAH
Penilaian kehadiran dan keterukan serangan ulat harimau telah dijalankan
secara rawak ke atas lokasi tanaman tongkat ali di Semenanjung Malaysia.
Secara amnya, ladang-ladang ini dimiliki oleh agensi kerajaan, sektor swasta
atau persendirian. Semasa pemantauan, gejala dan tanda serangan ulat
harimau direkodkan dan diambil gambar. Bagi rekod kehadiran dan keseriusan
serangan perosak tersebut, 30 pokok dipilih secara rawak di setiap ladang dan
kehadiran ulat harimau di rekod menggunakan formula berikut:
126
Kehadiran ulat harimau (%) = Bilangan pokok diserang ulat harimau x 100
Jumlah bilangan pokok yang dinilai
Tahap keseriusan serangan ulat harimau di kelaskan berdasarkan peratus daun
yang dimakan oleh ulat harimau mengikut kategori berikut; 0: tiada serangan,
1: <25% silara pokok dimakan, 2: 25% - 50% silara pokok dimakan, 3; 50% 75% silara pokok dimakan dan 4: >75% silara pokok dimakan. Indeks Keterukan
kerosakan (DSI) diperolehi menggunakan formula berikut:
Di mana DSI = Indeks keterukan kerosakan (Damage Severity Index)
no = Bilangan pokok dengan keterukan kategori 0
n1 = Bilangan pokok dengan keterukan kategori 1
n4 = Bilangan pokok dengan keterukan kategori 4
C = Kategori keterukan tertinggi
Kajian ini melibatkan sejumlah 28 lokasi tanaman tongkat ali yang dipantau.
Daripada jumlah ini, 11 lokasi merupakan petak tongkat ali yang ditubuhkan di
dalam hutan, 9 lokasi adalah ladang tongkat ali dengan selingan tanaman
pertanian atau pokok hutan dan 6 lokasi adalah ladang monokultur tongkat ali
(Rajah 1). Kesemua petak tongkat ali di dalam hutan adalah di bawah
pengurusan Jabatan Hutan Negeri bertujuan untuk pemuliharaan daripada
kepupusan spesies tersebut. Spesies yang ditanam secara selingan dengan
tongkat ali adalah jati, karas, akasia dan herba seperti serai wangi.
Pemerhatian juga mendapati bahawa, secara purata pokok tongkat ali di dalam
hutan mempunyai ketinggian kurang daripada 3 m dengan usia 1 hingga 5
tahun. Manakala, ketinggian pokok tongkat ali di ladang monokultur dan
selingan adalah di antara 1 m hingga 10 m dengan usia 5-10 tahun. Ladang
monokultur dan selingan menunjukkan saiz yang lebih besar di antara 2 hektar
ke 12.5 hektar berbanding tanaman di kawasan hutan iaitu 0.5 hektar hingga 2
hektar. Kebanyakan ladang tongkat ali yang dipantau terbiar dan tidak urus
dengan baik. Kebanyakan pemilik menyatakan bahawa aktiviti-aktiviti
penyelenggaraan seperti membaja, merumput dan meracun hanya dijalankan
pada peringkat awal penubuhan ladang sahaja.
127
Petunjuk:
Tanaman di kawasan hutan
Tanaman secara selingan
Tanaman secara monokultur
Rajah 1. Lokasi kawasan tanaman tongkat ali yang terlibat di dalam survei
serangan ulat harimau
Keputusan kajian ini mendapati kesemua lokasi tanaman tongkat ali
(100%) telah diserang oleh ulat harimau (Rajah 2). Kerosakan akibat serangan
perosak ini adalah pelbagai mengikut lokasi dengan julat daripada peringkat
minor kepada peringkat serius. Walau bagaimanapun, serangan yang teruk
mampu memusnahkan keseluruhan daun muda dan menyebabkan pokok
menjadi renggeh. Secara purata tongkat ali di kawasan hutan mencatatkan
sebanyak 16.67% kehadiran ulat harimau dan indeks keterukan serangan (DSI)
adalah 32.69%. Hampir kesemua petak tanaman tongkat ali di dalam hutan
menunjukkan kurang daripada 20% kehadiran ulat harimau kecuali petak128
petak di Pengkalan Hulu dan Bukit Hijau. Di Pengkalan Hulu, kehadiran ulat
harimau adalah 30% manakala, di Bukit Hijau mencatatkan sehingga 83.33%
kehadiran perosak tersebut. Kerosakan pada tanaman tongkat ali di Bukit
Hijau amat serius dengan purata kerosakan pada bahagian silara tongkat ali
adalah melebihi 75% . Bilangan tongkat ali yang masih hidup juga berkurangan
kepada 90% dari bilangan anak benih yang ditanam.
Rajah 2. Peratus kehadiran dan kategori keterukan kerosakan ulat harimau ke
atas tongkat ali di Semenanjung Malaysia
Kehadiran ulat harimau di ladang secara selingan pula adalah di antara
3.33% hingga 80.00% dengan purata sebanyak 25.95%. DSI pula adalah 33.33%.
Kebanyakan ladang tongkat ali secara selingan tidak mengalami kerosakan
yang teruk akibat serangan perosak ini walaupun ladang tongkat ali di Taman
Impian Herba dan Jati, Melaka mencatatkan 80% kehadiran ulat harimau.
Pemerhatian mendapati tongkat ali di ladang ini mempunyai saiz yang lebih
besar dengan ketinggian melebihi 8 m. Selain itu, silaranya adalah besar dan
129
menyebabkan serangan ulat harimau tidak menunjukkan kesan yang ketara.
Sebaliknya, ladang monokultur menunjukkan nasib yang berbeza berbanding
petak tongkat ali di kawasan hutan. Kajian mendapati hanya satu ladang
monokultur tongkat ali merekodkan kehadiran ulat harimau yang rendah iaitu
di Felda Jengka 16 (16.67%). Ladang-ladang tongkat ali yang lain seperti di
Felda Sungai Tekam dan Sungai Menyala masing-masing mencatatkan
kehadiran ulat harimau sebanyak 86.7% dan 83.33 %. Kedua-dua kawasan ini
juga menunjukkan tahap kerosakan pada silara pokok (DSI) yang tinggi iaitu
melebihi 75%.
Secara amnya, ladang-ladang tongkat ali secara monokultur dan
selingan mencatatkan kadar kehadiran ulat harimau dan kerosakan yang lebih
tinggi berbanding tongkat ali di kawasan hutan. Kajian ini juga menunjukkan
bahawa masalah serangan ulat harimau tidak terhad kepada ladang tongkat ali
secara monokultur dan selingan sahaja tetapi turut berlaku di kawasan hutan.
Kepelbagaian spesies pokok dan jarak di antara pokok tongkat ali di petak
dalam hutan yang besar merupakan faktor yang menjadikan serangan ulat ini
tidak ketara berbanding di ladang yang mengamalkan tanaman secara
monokultur dan selingan.
Untuk mengawal perosak ini, pendekatan kawalan secara biologi
seperti penggunaan kulat Metarhizium sp. dan Beauveria bassiana dan
bakteria, Bacillus thuringiensis (Mohd Anuar 2003) didapati lebih sesuai
berbanding pendekatan racun kimia yang sangat beracun. Di samping itu,
pendekatan ini juga mampu mengekalkan kehadiran musuh alami ulat harimau
di ladang seperti pemangsa, Sycanus sp. dan parasitoid.
RUMUSAN
Serangan ulat harimau ke atas tongkat ali berlaku di semua kawasan tanaman
tongkat ali termasuk di hutan dan di ladang (selingan dan monokultur). Walau
bagaimanapun, ladang tongkat ali monokultur adalah lebih terdedah kepada
serangan ulat harimau berbanding ladang selingan dan petak yang ditubuhkan
di kawasan hutan. Langkah terbaik dalam mengatasi masalah ini adalah dengan
membuat pemantauan berkala supaya serangan pada peringkat awal dapat
kesan dan langkah pengawalan dan pencegahan yang sesuai dapat diambil.
130
RUJUKAN
Mohd Anuar, A. 2003. Some Notes On The Control of The Tiger Moth, Atteva
sciodoxa (Lepidoptera: Yponomeutidae) and Scale Insects on Tongkat
Ali (Eurycoma Longifolia). The 6th International Conference on Plant
Protection in the Tropics; 11─14th. August 2003; Kuala Lumpur. P 92.
Mohd Farid, A., Wan Muhd Azrul, W.A., Ong S.P., & Lee S.S. 2014. Destructive
Leaf Cutter (Tiger Moth) of Eurycoma longifolia. FRIM Technical
Information No. 73.
Mohd. Noh Hj., Mohd Ilham, A. & Fauziah, I. 2004. Ancaman-Ancaman Utama
Dalam Penanaman Tongkat Ali Di Semenanjung Malaysia. Pp 42─46 in
Chang Y.S, Vimala, S., Mazura, M.P. & Ong, B.K. (Eds.). Proceeding of
the Seminar on Medicinal Plants: Tongkat Ali, Kacip Fatimah and
Pegaga - New Dimension in Complementary Healthcare. Kuala Lumpur.
Patahayah, M., Lee, S.S. & Mohd Farid, A. 2011. Penyakit, Perosak Dan
Gangguan Tanaman Tongkat Ali. FRIM Technical Information Handbook
No.41.
131
PRODUK HERBA DAN TREND PENCEMARAN MIKROORGANISMA
Y Norulaiman, BK Ong, MA Nor Azah, MH Nurhazwani, A Nor Hayati & MS
Amira Rina Nurdiana
Tel: 03-62797358 Faks: 03-62729805 E-mel: [email protected]
ABSTRAK
Pencemaran mikroorganisma di dalam penghasilan produk herba merupakan
salah satu masalah yang sering dihadapi oleh industri herba. Pencemaran ini
boleh berlaku di peringkat bahan mentah, pemprosesan, pembungkusan dan
juga semasa penyimpanan. Di FRIM, Makmal Kawalan Kualiti Hasilan Semula
Jadi (NPQC) menawarkan perkhidmatan Ujian Pencemaran Mikroorganisma.
Makmal NPQC telah menjalankan ujian pencemaran mikroorganisma ini,
terhadap pelbagai jenis persediaan herba yang terdiri daripada pelbagai
spesies dan bahagian tumbuhan herba. Jenis persediaan herba boleh
dikelaskan kepada 3 kategori iaitu; bahan mentah, separa siap dan produk siap.
Untuk setiap jenis persediaan herba spesifikasi atau had pencemaran
mikroorganisma yang ditetapkan oleh pihak berwajib adalah berbeza. Hasil
daripada ujian pencemaran yang telah dijalankan dapat diperhatikan pola atau
corak pencemaran mikroorganisma adalah berbeza untuk setiap kategori
persediaan herba. Kategori bahan mentah herba menunjukkan kadar
pencemaran yang paling tinggi berbanding separa siap dan produk siap. Pola
pencemaran mikroorganisma menunjukkan kadar pencemaran semakin
berkurang dengan bertambahnya proses yang dijalankan, bermula dari
pemprosesan bahan mentah ke peringkat produk siap. Walaubagaimanapun,
tahap pencemaran mikroorganisma yang lebih tinggi dapat diperhatikan untuk
sesetengah herba yang digunakan dari bahagian rizom/akar atau akar dalam
kategori separa siap dan produk siap berbanding bahan mentah herba,
walaupun telah melalui beberapa peringkat proses pengekstrakan. Ini
menunjukkan bahawa, pemilihan bahan mentah yang bersih dan berkualiti
amat penting dalam memastikan produk akhir yang dihasilkan adalah selamat
untuk digunakan.
Kata kunci: pencemaran mikroorganisma, bahan mentah, separa siap, produk
siap
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PENGENALAN
Pencemaran mikroorganisma ke dalam produk herba boleh mendatangkan
kemudaratan kepada pengguna. Mikroorganisma di dalam sesuatu produk
berupaya untuk membiak dan merembeskan toksin (Muller & Basedow 2007).
Pembiakan mikroorganisma di dalam produk, menyebabkan produk tersebut
rosak dengan bertukar warna, rupa fizikal serta memberikan bau yang tidak
menyenangkan (Maria et al. 1995). Ini menyebabkan jangka hayat sesuatu
produk herba ini menjadi lebih pendek dan terpaksa ditarik balik dari pasaran.
Bagi memasarkan produk herba, produk perlu didaftarkan dan melalui ujian
keselamatan. Salah satu ujian keselamatan adalah ujian pencemaran
mikroorganisma yang mana tahap pencemaran mikroorganisma akan dinilai
dan perlu mematuhi had spesifikasi yang ditetapkan. Untuk kajian ini,
persediaan herba dikelaskan kepada 3 kategori iaitu, bahan mentah herba,
produk separa siap dan produk siap. Bahan mentah herba adalah herba yang
telah dikeringkan dan dikisar dalam bentuk serbuk. Produk separa siap pula
merupakan herba yang telah melalui proses pengekstrakan dan juga campuran
bahan persediaan herba. Manakala produk siap merupakan produk herba yang
telah siap diproses dan dalam bungkusan akhir untuk dipasarkan seperti
kapsul, tablet herba dan produk penjagaan diri.
BAHAN DAN KAEDAH
Ujian Paras Kandungan Mikroorganisma (Microbial Enumeration Test)
Ujian dijalankan dengan menimbang 10 g sampel dan dicampurkan ke dalam
90 ml larutan sodium klorida pepton dan seterusnya sediaan pencairan bersiri.
1 ml daripada setiap pencairan dipiringkan ke dalam piring petri dan
dicampurkan dengan Agar Tryptik Soya (TSA) untuk pengiraan jumlah
mikroorganisma aerobik (TAMC) dan Sabouraud dekstrose agar (SDA) untuk
pengiraan jumlah yis dan kulat (TYMC). Campuran sampel dan agar yang telah
beku kemudiannya dieramkan dalam inkubator pada suhu 32.5°C selama 5 hari
untuk TSA dan campuran agar SDA dieramkan pada suhu 22.5°C sehingga 7
hari. Koloni mikroorganisma dikira dalam unit CFU/g@ml. (British
Pharmacopea, 2011)
Analisis dan Trend Paras Pencemaran Mikroorganisma
Keputusan ujian pencemaran mikroorganisma setiap sampel dibandingkan
dengan had spesifikasi yang telah ditetapkan oleh Biro Kawalan Farmaseutikal
Kebangsaan (BPFK). Analisis peratusan sampel yang memenuhi spesifikasi dan
trend paras pencemaran mikroorganisma mengikut kategori diwujudkan.
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Paras Pencemaran Mikroorganisma
Sebanyak 184 sampel bahan mentah, 66 produk separa siap dan 75 produk siap
telah dijalankan ujian pencemaran mikroorganisma. Dalam kategori bahan
mentah 64% adalah sampel daun, 21% adalah rizom/akar, 10% buah dan 5%
batang. Bagi kategori produk separa siap, 76% sampel adalah ekstrak dan 24%
adalah bahan persediaan herba. Manakala produk siap pula 65% adalah produk
untuk penggunaan oral dan 35% adalah produk untuk penggunaan topikal.
Hasil daripada ujian paras kandungan mikroorganisma didapati kategori bahan
mentah memberikan bacaan CFU/g@ml paling tinggi iaitu sehingga 1012
CFU/g@ml, berbanding kategori separa siap bacaan CFU/g@ml tertinggi adalah
sehingga 1010 CFU/g@ml dan produk siap sehingga 106 CFU/g@ml. Hasil
pemerhatian menunjukkan terdapat sampel herba dengan paras pencemaran
mikrooganisma melebihi had spesifikasi sehingga 10 x 103 kali ganda. Nilai
CFU/g@ml tertinggi iaitu 5.2 x 1011 CFU/g@ml adalah sampel kategori bahan
mentah daripada bahagian rizom. Bagi kategori separa siap; ekstrak herba
daripada bahagian rizom/akar dan buah mencatatkan nilai CFU/g@ml paling
tinggi iaitu 2.5 x 1010 CFU/g@ml. Manakala produk penggunaan topikal
memberikan bacaan yang paling tinggi berbanding produk penggunaan oral
iaitu 2.1 x 107 CFU/g@ml untuk kategori produk siap.
Analisis dan Trend Pencemaran Mikroorganisma
Peratusan sampel yang memenuhi had spesifikasi dapat dilihat seperti jadual 1.
Hampir 80% daripada sampel untuk semua kategori yang diuji memenuhi had
spesifikasi yang dietapkan untuk TAMC. Manakala hanya 50% memenuhi had
spesifikasi untuk TYMC.
Jadual 1. Keputusan paras kandungan mikroorganisma mengikut kategori
sediaan produk
Peratus
Bacaan Tertinggi
Had Spesifikasi
Kategori
Memenuhi
CFU/g@ml
CFU/g@ml
spesifikasi (%)
Bahan
52000 x 107
5 x 107 (TAMC)
90
5
5
Mentah
4400000 x 10
5 x 10 (TYMC)
51
2500000 x 104
5 x 104 (TAMC)
79
Separa Siap
2
2
88000 x 10
5 x 10 (TYMC)
59
600 x 104
5 x 104 (TAMC)
85
Produk Siap
210000 x 102
5 x 102 (TYMC)
57
134
Meneliti trend pencemaran mikroorganisma secara umum, jelas
bahawa paras pencemaran mikroorganisma pada produk kategori bahan
mentah herba adalah paling tinggi, diikuti produk kategori separa siap. Paras
pencemaran mikroorganisma paling rendah pada produk kategori siap (Rajah
1). Pola pencemaran ini dipercayai berlaku kerana sampel bahan mentah herba
yang dikutip dari ladang mempunyai hubungan langsung dengan sumber
pencemaran seperti bahagian rizom/akar dengan tanah dan bahagian
buah/daun yang dicemari oleh mikroorganisma. Oleh itu, adalah penting
proses pemilihan dan sanitasi bahan mentah herba dijalankan dengan segera
agar kontaminasi mikroorganisma tidak merebak dan semakin meningkat.
Proses sanitasi bahan mentah perlu dibuat dengan lebih rapi untuk bahan
mentah daripada bahagian rizom/akar dengan memastikan tanah dan kotoran
dibasuh bersih. Bahagian buah atau daun yang tercemar dengan
mikroorganisma juga perlu diasingkan.
Rajah 1. Paras pencemaran mikroorganisma mengikut kategori sediaan produk
Paras pencemaran mikroorganisma untuk kategori separa siap dan
produk siap adalah lebih rendah berbanding bahan mentah. Pola penurunan
paras mikroorganisma ini adalah kerana produk separa siap dan produk siap
telah melalui pelbagai peringkat pemprosesan. Proses pemilihan, sanitasi serta
pengeringan akan mengurangkan paras kandungan mikroorganisma pada
bahan mentah herba. Begitu juga dengan proses pengekstrakan yang
135
melibatkan penggunaan pelarut dalam penyediaan ekstrak dan juga haba
dalam penyediaan ekstrak air turut membantu mengurangkan paras
pencemaran mikroorganisma. Seterusnya proses pengeringan ekstrak seperti
kering beku (freeze dry) juga dapat mengelakkan pembiakan mikroorganisma
terutamanya kulat yang mudah membiak dalam keadaan yang lembap.
Walaubagaimanapun pencemaran mikroorganisma untuk kategori separa siap
dan produk siap ini boleh berlaku semula semasa peringkat pengendalian,
pembungkusan dan penyimpanan produk.
Jadual 2 menunjukkan peratus penurunan paras mikroorganisma yang
telah melalui proses pengekstrakan. Paras mikroorganisma kategori bahan
mentah, bahagian daun menunjukkan penurunan melebihi 75%, manakala
penurunan sehingga 33% ditunjukkan oleh bahagian rizom/akar. Jadual 2
menunjukkan paras pencemaran produk separa siap bahagian rizom/akar lebih
tinggi berbanding bahagian daun selepas melalui proses pengekstrakan.
Jadual 2. Penurunan paras mikroorganisma yang telah melalui pemprosesan
Bahan
Separa Siap/
Peratus
Bahagian
Sampel
Mentah
Produk Siap
Penurunan
Herba
Log 10
Log 10
(%)
CFU/g@ml
CFU/g@ml
LB
8.3
7.3
12.0
Rizom/akar
PT
7.0
4.7
33.3
KP
7.0
5.8
16.6
CK
6.9
4.8
31.3
DB
5.1
1.0
80.4
KR
9.4
2.0
79.2
Daun
KJ
7.1
1.6
77.4
KE
5.5
1.0
82.0
RUMUSAN
Pencemaran mikroorganisma umumnya paling tinggi dalam kalangan produk
herba kategori bahan mentah dan berkurangan pada produk siap (yang proses
penghasilannya mungkin melibatkan penggunaan haba dan pelarut). Namun,
tiada jaminan bahawa produk yang bakal terhasil mampu memenuhi spesifikasi
yang ditetapkan jika bahan mentah yang digunakan untuk penghasilan produk
siap telah tercemar dengan teruk. Adalah disyorkan perhatian yang khusus bagi
penghasilan produk herba dari bahagian rizom/akar, kerana sesetengah
mikroorganisma daripada tanah mempunyai ketahanan yang tinggi terhadap
haba dan pelarut yang digunakan pada pelbagai peringkat pemprosesan.
136
PENGHARGAAN
Penghargaan ditujukan kepada semua ahli Makmal Kawalan Kualiti Hasilan
Semula Jadi, serta kakitangan Program Pembangunan Produk Herba di
Bahagian Hasilan Semula Jadi atas bantuan yang diberikan. Juga kepada FRIM
atas bantuan dana dalam menjalankan ujian di Makmal NPQC.
RUJUKAN
Maria, C.R., Maria, R.M. & Carmen, V. (1995). Microbiological Quality of
Pharmaceutical Raw Materials. Pharmaceutica Acta Heltivitiae. 70:
227─232.
Microbial Enumeration Test. Pour Plate Method British Pharmacopoeia 2011:
Appendix XVI B. A430─A434 pp.
Muller, P., & Basedow, T. (2007). Aflatoxin Contamination of Pods of Indian
Cassia senna L. (Caesalpinaceae) Before Harvest, During Drying and In
Storage: Reason and Possible Methods of Reduction. Journal of Stored
Products Research. 43: 323─329.
137
RUMPAI MIANG MEXICO, ANCAMAN MELEBIHI MANFAAT
AK Azimah1, MA Anis Fadzilah1, M Dilipkumar2 & AH Muhammad Saiful3
Pusat Penyelidikan Hortikultur, Ibu Pejabat MARDI, Serdang
Pusat Penyelidikan Padi, Seberang. Perai, Pulau Pinang
ABSTRAK
Rumput miang mexico (Parthenium hysterophorus) dengan nama Inggerisnya
“Parthenium weed” atau “congress grass” merupakan rumpai berbahaya yang
merebak ke seluruh benua Asia, Australia, Eropah, Afrika, Amerika Utara dan
Selatan yang merangkumi sebanyak 82 buah negara termasuk Malaysia.
Rumput miang mexico mula dijumpai di Malaysia pada September 2013 di Ulu
Yam, Selangor. Rumput ini melengkapkan kitaran hidup dalam tempoh 4-5
minggu dan satu pokok tunggal mampu menghasilkan 15,000-100,000 biji
benih. Rumput ini menduduki tangga teratas rumpai paling berbahaya di dunia
kerana kesan buruknya kepada manusia, haiwan, tanaman dan ekosistem.
Antaranya boleh menyebabkan asma, bronkitis dan dermatitis kepada
manusia. Debunganya yang halus, bahagian tumbuhan yang kering dan akar
adalah bahagian yang berbahaya jika dibiarkan terdedah secara berterusan.
Mereka yang tergolong di kalangan kanak-kanak dan golongan berumur adalah
berisiko tinggi untuk mendapat kesan ini. Rumput miang mexico juga
menyebabkan toksik kepada haiwan terutamanya haiwan ternakan yang
terdedah kepadanya. Makanan ternakan dalam kadar 10-50% yang dicemari
dengan rumput miang mexico boleh menyebabkan aneroksia, diarrhea, sakit
mata atau kesakitan akut yang membawa maut. Rumpai ini turut mempunyai
kesan alelopati atau perencatan terhadap tumbuhan lain yang tumbuh di
sekitarnya dan boleh menyebabkan kehilangan hasil di kawasan pertanian jika
tidak dikawal.
Kata kunci: Parthenium hysterophorus, rumpai miang meixco, biji benih,
rumpai berbahaya, alelopati
PENGENALAN
Parthenium hysterophorus atau nama tempatannya rumpai miang mexico, juga
dikenali sebagai ‘Parthenium weed’, ‘congress grass’, ‘carrot weed’ dan ‘star
weed’ merupakan rumpai yang agresif dan tidak mempunyai nilai ekonomi
(Patel 2011) selain daripada bersifat berbahaya dan cukup tidak
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menyenangkan (Rezaul 2010). Rumpai ini berasal dari Mexico, Amerika tengah
dan Amerika Selatan. Rumpai daripada keluarga Asteraceae ini boleh tumbuh
di pelbagai jenis tanah termasuk tanah yang kurang subur.
Morfologi Rumpai Miang Mexico
Rumpai miang mexico seakan ulam raja pada peringkat muda, daunnya
berbulu halus dan tersusun membulat. Bunganya kecil bewarna putih krim
muncul berjambak di hujung pokok dan bertukar menjadi coklat apabila
matang. Batang biasanya bercabang dan diselaputi bulu halus atau dipanggil
trikom dengan ketinggian mencapai sehingga 1.5 hingga 2 m. Biji benih
berbentuk baji dengan warna perang kehitaman. Satu pokok yang sihat boleh
menghasilkan 10,000 hingga 15,000 biji benih yang mampu bercambah dalam
satu kitaran yang lengkap (Patel 2011).
Kebaikan dan Keburukan
Biji benihnya boleh merebak melalui air mengalir, angin, makanan ternakan
yang dicemari, kenderaan dan jentera, haiwan ternakan yang diimport dan
manusia. Seluruh pokok termasuk debunga dan trikom mempunyai bahan
kimia toksin yang dipanggil sesquiterpen lakton dan pelbagai jenis bahan
alelokimia (Maishi et al., 1998). Bahan kimia ini boleh menyebabkan keburukan
kepada manusia, haiwan, tumbuhan dan ekosistem. Antaranya ia boleh
menyebabkan asma, bronkitis dan dermatitis kepada manusia. Kadar 10-50%
parthenium yang tercemar di dalam makanan ternakan boleh menyebabkan
kesakitan akut dan membawa kepada kematian. Anjing yang terdedah dengan
tanaman ini turut menyebabkan aneroksia, diarrhea dan sakit mata. Kehadiran
bahan kimia seperti parthenin, hysterin, hymenin dan ambrosin telah memberi
kesan alelopati atau perencatan terhadap tumbuhan lain yang tumbuh di
sekitarnya. Kajian telah membuktikan bahawa parthenin adalah bahan kimia
yang boleh merencat pertumbuhan akar pelbagai jenis tumbuhan dikot dan
monokot. Keistimewaan ini membolehkannya mudah menguasai sesuatu
kawasan dan menyesarkan tanaman lain. Apabila rumpai agresif ini menempati
sesuatu kawasan, maka biodiversiti kawasan tersebut akan berubah.
Perubahan drastik ini pasti menyebabkan ketidakseimbangan ekosistem
berlaku dan perlu ditangani dengan segera.
Namun demikian, terdapat suku kaum tertentu memanfaatkan rumpai
ini sebagai ramuan untuk mengubati radang, ekzema, ruam kulit, sakit sendi,
selesema, penyakit hati dan pelbagai lagi (Patel 2011). Kajian oleh Das et
al.(2007) menunjukkan bunganya yang diekstrak mempunyai bahan antitumor,
manakala Patel et al. (2008) membuktikan bunga rumpai ini mampu mengawal
kencing manis.
139
Oleh kerana telah tersebar luas tentang keburukan rumpai ini kepada
orang awam, maka kajian telah dijalankan untuk mengenalpasti taburan
rumpai miang mexico di beberapa tempat yang telah dilaporkan terdapat di
Malaysia. Di samping itu, kajian turut menekankan jenis tanah bagaimanakah
rumpai ini paling senang ditemui.
BAHAN DAN KAEDAH
Kajian dijalankan di tiga tempat yang telah dilaporkan merebak seperti di Ulu
Yam, Selangor; Sungai Siput, Perak dan Sungai Pasir, Kedah. Pemantauan yang
dijalankan meliputi beberapa aspek termasuk keluasan kawasan yang telah
ditumbuhi, jenis tanah, faktor persekitaran yang memungkinkan berlakunya
penyebaran dan jenis aktiviti kawasan tersebut.
Ujian Percambahan Biji Benih
Kajian ini bertujuan untuk mengenal pasti keupayaan biji benih bercambah
pada tanah yang berbeza. Biji benih dan tanah telah diambil dan ujian
percambahan telah dilakukan di atas piring petri dan juga di atas tanah. Biji
benih telah direndam dengan bahan kimia kalium nitrate (0.2%) dan juga air
yang telah ditapis untuk memecahkan dormansi selama 24 jam. Kemudian biji
benih tersebut telah ditos selama 12 jam sebelum disemai di atas piring petri
dan tiga jenis tanah yang berbeza. Selepas dua minggu, pemerhatian dan
keputusan percambahan telah diambil.
Soal Selidik Kesan Rumpai kepada Persekitaran
Soal selidik telah dijalankan dengan menemu ramah beberapa orang pegawai
pengembangan di kawasan tersebut. Soalan merangkumi kes terhadap
kesihatan manusia, haiwan ternakan dan tanaman. Kes adalah berdasarkan
kepada laporan yang disampaikan kepada pegawai terlibat.
140
Ujian percambahan biji benih
Analisis tekstur tanah (Jadual 1) menunjukkan bahawa tanah dari kawasan Ulu
yam adalah berpasir manakala tanah dari Sg. Pasir adalah lempung liat dan Sg.
Siput adalah lempung.
Rajah 1 menunjukkan ujian percambahan biji benih rumpai miang
mexico telah dibuat dengan rawatan 0.2% kalium nitrat (KNO3) dan air suling.
KNO3 digunakan sebagai bahan perangsang percambahan bagi biji benih P.
Hysterophorus. Keputusan menunjukkan percambahan biji benih yang
direndam dalam air suling lebih tinggi berbanding dengan yang dirawat dengan
KNO3. Ini membuktikan bahawa biji benih P. hysterophorus mempunyai
keupayaan untuk bercambah dengan air biasa dan biji benihnya mampu
bercambah tanpa rangsangan dari bahan kimia yang lain. Kajian yang dilakukan
oleh Adkins et al. (2010) menunjukkan biji benih P. hysterophorus mempunyai
keupayaan untuk bercambah dan tumbuh dengan sendiri dan keupayaan ini
boleh mencapai tahap maksima mengikut suhu dan jenis tanah. Ujian
percambahan biji benih pada media tanah dan piring petri juga telah dilakukan
dan ujian mendapati bahawa kadar percambahan biji benih pada media tanah
lebih tinggi berbanding piring petri (Rajah 2). Tanah dari kawasan Sg. Siput
mempunyai kadar percambahan yang paling tinggi berbanding dengan tanah
dari kawasan Ulu yam dan Sg. Pasir (Rajah 3).
Jadual 1. Analisis tekstur tanah di tiga tempat yang berbeza
Tempat
asal biji
benih
Ulu
Yam
Sg Pasir
Taburan saiz partikel (mikron)
clay
Silt
0-2
2.5
220.0
4.47
33.4
19.06
Jumlah mikron
fine
sand
medium
sand
coarse
sand
2050
2.23
very
fine
sand
50100
3.94
Total
100250
19.28
250-500
>500
28.42
39.12
99.97
6.99
5.74
13.96
9.99
10.88
99.98
Kelas
tekstur
tanah
Clay
Silt
Sand
<2
>50
USDA
2.5
250.0
6.7
90.76
Pasir
33.35
26.05
40.57
Lempung
liat
Sg
Siput*
Lempung
Nota: * Ditentukan berdasarkan pemerhatian terhadap tanah bukan
berdasarkan analisis tanah
141
Rajah 1. Kadar percambahan biji benih di atas tanah dan piring petri selepas
rawatan dengan 0.2% KNO3
Rajah 2. Kadar percambahan biji benih P. hysterophorus selepas rawatan
dengan air suling
142
Rajah 3. Perbezaan kadar percambahan biji benih P. hysterophorus selepas
rawatan dengan KNO3 dan air suling
Soal Selidik Kesan Rumpai kepada Persekitaran
Pemantauan telah dibuat di kawasan tersebut mendapati kawasan Ulu Yam
dan Sg. Siput merupakan kawasan penternakan lembu dan rusa. Kawasan Ulu
Yam merupakan salah satu tapak kekal pengeluaran makanan (TKPM). Hampir
keseluruhan kawasan tersebut telah tersebar dengan pertumbuhan yang
berkelompok dan tidak sekata. Kebanyakkannya tumbuh di tepi jalan dan
kawasan penternakan. Kawasan Sg. Siput pula merupakan ladang infoternak
yang menempatkan ternakan rusa dan lembu. Penyebaran rumpai miang
mexico adalah sekata dan seragam di keseluruhan kawasan ladang. Walau
bagaimanapun, sehingga tarikh tinjauan dilakukan, masih belum terdapat kes
yang berkaitan rumpai tersebut dilaporkan kepada pihak berwajib. Kebanyakan
ternakan tidak meragut rumpai tersebut dan mengelak memakannya. Pekerja
yang bertugas di kawasan tersebut juga tidak mengalami sebarang alahan dan
simptom berkaitan rumpai berkenaan. Namun jika dilihat kepada hasil
tanaman, kawasan tersebut telah menyebabkan kos pengawalan meningkat
apabila banyak herbisid yang terpaksa digunakan untuk mengawalnya. Hasil
tanaman turut terjejas ekoran pertumbuhan rumpai yang sangat agresif
(Tefera 2002). Kemungkinan juga, kualiti hasil tanaman akan berkurangan dan
tercemar dengan biji benih rumpai ini. Ekosistem turut berubah apabila terlalu
banyak rumpai miang mexico menempati kawasan tersebut sehingga
menyebabkan biodiversiti turut terganggu (Kohli et al. 2006).
143
KESIMPULAN
Kajian ini membuktikan biji benih P. hysterophorus yang diambil dari tiga
kawasan yang berbeza di Semenanjung Malaysia mempunyai keupayaan yang
tinggi untuk bercambah walaupun pada tekstur tanah berbeza yang berbeza.
Walaupun kesan buruk tumbuhan terhadap manusia dan haiwan masih boleh
dikawal, namun kesannya terhadap tanaman dan ekosistem telah jelas berlaku.
Oleh kerana itu, pengawalan dan pemantauan yang serius perlu diambil oleh
semua pihak agar rumpai ini tidak terus tersebar.
PENGHARGAAN
Kami mengucapkan berbanyak terima kasih kepada pihak pengurusan dan
pegawai pengembangan Ladang Infoternak, Sg. Siput Perak dan Ladang TKPM
Ulu yam, Selangor atas kerjasama memberikan maklumat tentang rumpai
miang mexico dan semua pihak yang terlibat secara langsung atau tidak
langsung menyiapkan artikel ini.
RUJUKAN
Adkins, S. W., O’Donnell, C., Khan, N., Nguyen, T., Shabbir, A., Dhileepan, K.,
George, D. & Navie, S. (2010). Parthenium Weed (Parthenium
hysterophorus L.) Research in Australia: New Management Possibilities.
pp. 120-123 in Zydenbos, S.M.(ed) Proceedings 17th Australasian
Weeds Conference.
Das, B., V. S. Reddy, M. Krishnaiah, A. V. S. Sharma, K. R. Kumar, J. V. Rao & V.
Sridhar (2007). Acetylated Pseudoguaianolides from Parthenium
Hysterophorus and Their Cytotoxic Activity. Phytochemistry 68 (15):
2029─2034.S
Kohli, R. K., Batish, D. R., Singh, H. P., & Dogra, K. S. (2006). Status, Invasiveness
and Environmental Threats of Three Tropical American Invasive Weeds
(Parthenium hysterophorus L., Ageratum conyzoides L., Lantana
camara L.) in India. Biological Invasions, 8(7), 1501─1510.
Maishi, A. I., Ali, P. S., Chaghtai, S. A., & Khan, G. (1998). A Proving of
Parthenium hysterophorus, L. British Homoeopathic Journal, 87(1),
17─21.
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Patel, S. (2011). Harmful and Beneficial Aspects of Parthenium hysterophorus:
an Update. 3 Biotech, 1(1), 1─9.
Patel, V. S., Chitra, V., Prasanna, P. L., & Krishnaraju, V. (2008). Hypoglycemic
Effect of Aqueous Extract of Parthenium hysterophorus l. in Normal and
Alloxan Induced Diabetic Rats. Indian journal of pharmacology, 40(4),
183.
Rezaul, S.K. (2010). Ill Impacts of Parthenium Weed on Human Health,
Livestock Production and Environment.
Tefera, T. (2002). Allelopathic Effects of Parthenium Hysterophorus Extracts on
Seed Germination and Seedling Growth of Eragrostis tef. Journal of
Agronomy and Crop Science, 188(5), 306─310.
145
KONSERVASI SECARA EX SITU BAGI KACIP FATIMAH VARIETI
LANCEOLATA
SB Syafiqah Nabilah, MA Farah Fazwa, S Norhayati & M Nur Nazihah
Institut Penyelidikan Perhutanan Malaysia, 52109 Kepong, Selangor
Tel: 03-6279 7332
Faks: 03-6280 4614
ABSTRAK
Kacip fatimah atau nama saintifiknya Labisia pumila ialah sejenis tumbuhan
dalam famili Primulaceae yang digunakan oleh masyarakat Melayu sejak 400
tahun yang lalu. Pokok kacip fatimah adalah sejenis tumbuhan renek yang
tumbuh secara menegak dengan anggaran ketinggian 30–50 cm. Pokok ini
mempunyai akar yang unik, iaitu akar tunjang yang panjang dan dikelilingi oleh
anak akar yang halus. Malahan, anak pokok kacip fatimah boleh tumbuh keluar
di sepanjang akarnya itu. Hasil daripada kajian taksonomi botani, terdapat tiga
jenis varieti kacip fatimah yang biasa dijumpai di Malaysia iaitu Labisia pumila
varieti alata, Labisia pumila varieti pumila dan Labisia pumila varieti
lanceolata. Proses pengecaman bagi varieti lanceolata mudah dilakukan
berbanding varieti pumila dan varieti alata kerana tangkainya tidak
mempunyai sayap. Walau bagaimanapun, habitat bagi varieti lanceolata
tumbuh lebih berselerak berbanding dua varieti yang lain yang tumbuh secara
berumpun. Setakat ini, kajian saintifik mengenai kegunaan dan kelebihan
varieti lanceolata adalah masih kurang berbanding dua varieti yang lain.
Sebagai langkah awal, Program Membaikbiak Tumbuhan, Institut Penyelidikan
Perhutanan Malaysia (FRIM) telah mengambil inisiatif untuk melakukan
konservasi secara ex situ bagi varieti ini untuk tujuan penyelidikan dan koleksi
germplasma. Dalam kajian ini, pokok-pokok ibu varieti lanceolata telah dikutip
dari lima hutan simpan di lima negeri iaitu Perak, Negeri Sembilan, Johor,
Terengganu dan Pahang. Selain pokok, sampel tanah bagi setiap lokasi juga
turut diambil bagi mengenalpasti kandungan nutrien yang membantu
pertumbuhan pokok ini di habitat asalnya. Pokok-pokok ibu yang dibawa
pulang dibiarkan pulih di tapak semaian sebelum ianya ditanam ke plot
germplasma. Pembiakan tampang menggunakan kaedah keratan juga turut
dilakukan untuk memperbanyakkan anak pokok. Keupayaan pengakaran varieti
lanceolata bagi populasi Perak telah direkodkan di dalam kajian ini. Hasil kajian
menunjukkan 80% keratan dari 12 genotip bagi populasi Perak telah berjaya
berakar dalam tempoh 9 minggu. Kadar pertumbuhan pucuk pada keratan
akar juga didapati lebih cepat berbanding keratan daun, di mana keratan yang
berakar telah mula mengeluarkan pucuk pada minggu ketiga selepas keratan
berakar dipindahkan ke dalam polibeg. Anak-anak pokok yang terhasil dari
kaedah keratan kemudiannya dipindahkan ke tapak semaian bagi tujuan
146
pengikliman dan penyesuaian sebelum ditanam di germplasma. Kadar
pertumbuhan pokok di germplasma juga direkodkan setiap bulan. Sebagai
kesimpulannya, pokok-pokok kacip fatimah yang ada di germplasma ini
digunakan sebagai bahan penyelidikan (breeding materials) khususnya dalam
bidang pembiakbakaan dan juga sebagai pusat rujukan bagi spesies ini. Selain
itu, populasi kacip fatimah di hutan simpan semula jadi juga dapat dilindungi
dan dipelihara.
Kata kunci: germplasma, keratan, pengakaran, pertumbuhan pucuk, breeding
materials, pemeliharaan
PENGENALAN
Malaysia terkenal dengan kekayaan sumber biodiversiti yang terdiri daripada
pelbagai spesies flora dan fauna. Tumbuhan herba merupakan salah satu
daripadanya. Lebih daripada 2,000 spesies tumbuhan dilaporkan mempunyai
ciri-ciri penyembuhan (healing qualities) dan berpotensi tinggi untuk
dikomersialkan (Saari & Noraini 2013). Kacip fatimah merupakan salah satu
tumbuhan herba tempatan yang mempunyai nilai komersial yang tinggi dan
sering mendapat permintaan di kalangan pengusaha-pengusaha herba
tempatan. Perkembangan kajian saintifik terhadap kacip fatimah telah
meningkatkan lagi keyakinan pengguna herba dan pengamal perubatan
tradisional. Selain digunakan sebagai air rebusan atau ubat periuk oleh wanita
bersalin, ianya juga boleh digunakan untuk merawat senggugut dan sakit sendi
di kalangan wanita menopaus (Intan & Nik 2015). Kajian–kajian sains yang
terkini pula telah membuktikan bahawa kacip fatimah berpotensi sebagai agen
anti-penuaan (Hyun-kyung et al. 2010), anti-obesiti (Fatilah et al. 2013) dan
anti-fungal (Ehsan et al. 2011). Oleh yang demikian, permintaan terhadap kacip
fatimah dijangka akan lebih meningkat dari tahun ke tahun. Walau
bagaimanapun, bekalan bahan mentah bagi spesies ini masih kerap dikutip dari
sumber hutan dan sedikit dari sumber perladangan. Kepupusan sumber bahan
mentah kacip fatimah boleh terjadi sekiranya tiada usaha pemuliharaan di
lakukan ke atas spesies ini. Justeru itu, kajian ini dijalankan dengan
mengetengahkan kaedah konservasi secara ex situ bagi spesies kacip fatimah
dari varieti lanceolata yang dijalankan oleh penyelidik dari Program
Membaikbiak Tumbuhan, Institut Penyelidikan Perhutanan Malaysia (FRIM).
147
BAHAN DAN KAEDAH
Pengumpulan Sampel Pokok dan Tanah
Lebih dari 150 genotip kacip fatimah varieti lanceolata telah dikutip dari lima
hutan simpan (HS) di Semenanjung Malaysia iaitu HS Gunung Korbu, Perak; HS
Gunung Berembun, Negeri Sembilan; HS Gunung Berlumut, Johor; HS Pulau
Selimbar, Terengganu; dan HS Betung, Pahang. Setiap pokok dilabel dengan
kod yang berbeza dan dibawa pulang ke FRIM. Sampel tanah bagi setiap
populasi kacip fatimah yang dikutip turut diambil untuk dianalisa di Makmal
Kimia Tanah. Antara analisis yang dijalankan adalah penentuan jumlah nitrogen
(%), fosforus (ppm) dan potassium (cmol/kg).
Pemulihan Pokok Di Tapak Semaian
Pokok-pokok kacip fatimah yang dibawa pulang ditanam semula di dalam
polibeg yang mengandungi media tanaman tanah: kompos: pasir (3:2:1) untuk
tujuan pemulihan. Pokok-pokok diletakkan di bawah lindungan 60% cahaya
mengikut populasi asal masing-masing dan disiram tiga kali sehari. Proses
pemulihan ini mengambil masa lebih kurang empat minggu sebelum ianya
dipindahkan ke germplasma.
Pembiakan Melalui Kaedah Keratan Akar
Dalam kajian ini, keratan akar telah digunakan untuk memperbanyakkan anak
pokok kacip fatimah. Akar dikerat sepanjang 5 cm dan hormon penggalak akar
diletakkan di bahagian bawah keratan. Keratan akar ditanam di dalam media
pasir dan disiram menggunakan sistem renjisan automatik selama satu minit
setiap jam. Peratus pengakaran direkodkan dalam tempoh 9 minggu.
Penubuhan Germplasma
Pokok-pokok ibu kacip fatimah yang telah dipulihkan dipindahkan ke plot
germplasma kacip fatimah yang sedia ada di FRIM. Plot germplasma yang
berkeluasan 0.4 hektar ini juga mempunyai lebih daripada 300 klon kacip
fatimah dari varieti alata dan pumila. Pokok-pokok kacip fatimah varieti
lanceolata ditanam mengikut populasi masing-masing dan dilabel mengikut
kod yang telah ditetapkan.
148
Bancian Pertumbuhan Pokok Di Germplasma
Bancian pertumbuhan seperti tinggi pokok (cm) dan diameter kolar (mm)
direkodkan setiap bulan. Analisis statistik telah dijalankan untuk mengenalpasti
perbezaan signifikan pertumbuhan pokok bagi setiap populasi.
Hasil Analisa Tanah
Analisa sampel tanah dari tiga populasi iaitu HS Gunung Korbu, Perak; HS
Gunung Berembun, Negeri Sembilan; dan HS Gunung Berlumut, Johor
direkodkan. Keputusan analisa makronutrien tanah bagi ketiga-tiga populasi
tersebut direkodkan di dalam Jadual 1.
Jadual 1. Sifat kimia tanah di habitat asal kacip fatimah varieti lanceolata
Populasi
Jumlah N % F
K
pH kering
(ppm)
(cmol/kg)
HS Gunung
0.36a ± 0.03 12.57a ± 1.88 0.12a ± 0.01
4.05b ± 0.04
Korbu
HS Gunung
0.25b ± 0.01 2.42b ± 0.19 0.11a ± 0.01
4.23a ± 0.04
Berlumut
HS Gunung
0.33a ± 0.02 4.49b ± 0.31 0.12a ± 0.01
4.08b ± 0.05
Berembun
Hutan Simpan Gunung Korbu, Perak mencatatkan jumlah makronutrien
tertinggi; jumlah nitrogen (0.36 %) dan fosforus tersedia (12.57 ppm) diikuti
dengan HS Gunung Berembun, Negeri Sembilan; (0.33%) dan fosforus tersedia
(4.49 ppm). Jumlah makronutrien yang rendah dicatatkan pada HS Gunung
Berlumut, Johor; jumlah nitrogen (0.25%) dan fosforus tersedia (2.42 ppm)
Walaubagaimanapun bacaan potassium, K bagi ketiga-tiga populasi tidak
menunjukkan perbezaan signifikan di mana julatnya adalah 0.11-0.12 cmol/kg.
Bacaan pH kering tanah bagi ketiga-tiga populasi pula diklasifikasikan sebagai
sangat berasid iaitu dalam julat 4.0–4.5 (The Potash Development Association
2011.)
Bagi ciri-ciri fizikal tanah pula (Jadual 2), HS Gunung Berembun, Negeri
Sembilan mencatatkan peratusan pasir kasar yang tinggi (45.90%) berbanding
pasir halus (11.30%). Manakala HS Gunung Berlumut, Johor pula mengandungi
pasir halus yang tinggi dan kelodak dengan masing-masing berjumlah 23.81%
dan 19.00%. Hutan Simpan Gunung Korbu, Perak pula mencatatkan kandungan
149
tanah liat yang tinggi (41.53%) diikuti dengan HS Gunung Belumut, Johor
(38.38%). Ciri fizikal tanah memainkan peranan penting kerana ia memberikan
kesan terhadap pertumbuhan pokok terutamanya keupayaan memegang air,
pengudaraan dan pengekalan bahan organik (Fisher & Binkley 2000).
Jadual 2. Ciri-ciri fizikal tanah di habitat asal kacip fatimah varieti lanceolata
Populasi
Pasir kasar
Pasir halus
Kelodak
Tanah liat
(%)
(%)
(%)
(%)
HS Gunung
36.06b ± 1.78
20.13ab ± 6.76 10.00b ± 0 .49 41.53a ± 1.38
Korbu
HS Gunung
37.5b ± 3.94
23.81a ± 2.38
19.00a ± 1.92 38.38a ± 16.96
Berlumut
HS Gunung
45.9a ± 1.94
11.13b ± 1.50
13.47b ± 0.69 31.40a ± 2.99
Berembun
Prestasi Keratan Akar
Peratus pertumbuhan pucuk (%)
Peratus pertumbuhan akar (%)
Bagi analisis prestasi keratan akar pula, hanya 12 genotip kacip fatimah var.
lanceolata (AL8, AL10, AL22, AL26, AL27, AL32, AL34, AL36, AL41, AL44, AL48
dan AL52) dari HS Gunung Korbu, Perak direkodkan dalam kajian ini. Selepas
sembilan minggu keratan, kebanyakan genotip yang diuji menunjukkan 80%
pengakaran dan hanya tiga genotip yang berjaya memberikan 100%
pengakaran iaitu AL10, AL32 dan AL36. Berdasarkan Rajah 1, genotip AL34
menunjukkan pertumbuhan akar yang cepat pada minggu ketiga diikuti dengan
genotip dari AL8, AL10, AL32, AL36, AL41 dan AL44.
60
50
50
40
40
30
30
20
20
10
10
0
0
Genotip
Rajah 1. Peratus pengakaran keratan
akar pada minggu ketiga
Genotip
Rajah 2. Peratus pertumbuhan pucuk
selepas tujuh minggu di polibeg
Selepas sembilan minggu, keratan akar yang berakar dipindahkan ke
dalam polibeg. Pertumbuhan pucuk pada keratan akar boleh dilihat selepas
tiga minggu keratan dialihkan ke dalam polibeg. Berdasarkan Rajah 2, genotip
AL32 menunjukkan pertumbuhan pucuk yang cepat berbanding genotip lain di
150
mana kebanyakannya mula mengeluarkan pucuk pada minggu ketujuh. Aminah
(2005) dan Rozihawati (2008) melaporkan pertumbuhan pucuk pada keratan
akar adalah lebih cepat berbanding keratan daun. Maklumat ini adalah penting
untuk pengusaha ladang yang ingin membiakkan tanaman kacip fatimah secara
konvensional.
Pertumbuhan Pokok Di Germplasma
Prestasi pertumbuhan pokok-pokok ibu dari lima populasi kacip fatimah varieti
lanceolata yang telah ditanam di germplasma direkodkan setiap bulan. Hasil
analisis statistik (ANOVA) menunjukkan prestasi pertumbuhan lima populasi
kacip fatimah varieti lanceolata menunjukkan perbezaan yang signifikan pada
p<0.05. Min ketinggian dan diameter pokok bagi setiap populasi selepas tiga
bulan ditanam direkodkan dalam Jadual 3.
Jadual 3. Prestasi pertumbuhan lima populasi kacip fatimah varieti lanceolata
di plot germplasma, FRIM
Populasi
Tinggi pokok (cm)
Diameter kolar (mm)
HS Gunung Korbu
13.17ab ± 0.73
4.95a ± 0.16
HS Gunung Berlumut
12.08b ± 0.38
4.43b ± 0.07
HS Gunung Berembun
13.63a ± 0.33
5.03a ± 0.07
HS Betung
9.85c ± 0.60
5.05a ± 0.17
HS Pulau Selimbar
9.28c ± 0.42
4.36b ± 0.09
Berdasarkan Jadual 3, pertumbuhan kacip fatimah varieti lanceolata dari
populasi HS Gunung Berembun menunjukkan prestasi pertumbuhan yang
paling bagus dengan ketinggian 13.63a ± 0.33 cm dan diameter kolar 5.03a ±
0.07 mm. Manakala pokok-pokok dari populasi HS Pulau Selimbar mencatatkan
prestasi pertumbuhan terendah dengan ketinggian 9.28c ± 0.42 cm dan
diameter kolar 4.36b ± 0.09 mm. Perbezaan dari segi pertumbuhan ini adalah
berkemungkinan disebabkan perbezaan umur pokok yang tidak diketahui
semasa dikutip dari kawasan hutan semula jadi.
RUMUSAN
Maklumat dari hasil kajian ini boleh digunapakai untuk tujuan pemuliharaan
dan pemeliharaan spesies kacip fatimah khususnya dan juga bagi spesiesspesies herba yang lain. Kesimpulannya, kacip fatimah varieti lanceolata juga
boleh diperbanyakkan menggunakan kaedah keratan sama seperti varieti alata
dan pumila. Para petani dan pengusaha herba boleh menggunakan teknik ini
untuk memperbanyakkan bahan tanaman di masa akan datang tanpa perlu
151
bergantung sepenuhnya daripada sumber hutan bagi mengawal dari
berlakunya kepupusan, dengan itu keselamatan diversiti spesies akan terjamin.
RUJUKAN
Aminah, H., Naimah, C.L., Mohd Zaki, A. & Lokmal, N. (20079. Rooted Leaf
Cuttings of Labisia pumila. Journal of Tropical Medicinal Plants: 9(1):
593─599.
Ehsan, K., Z. E. J. Hawa & A. Sahida,. (2011). Phytochemical Analysis and
Antimicrobial Activities of Methanolic Extracts of Leaf, Stem and root
From Different Varieties of Labisia pumila Benth. Molecules: 16:
4438─50.
Fathilah, S.N., Norazlina, M., Norliza, M., Isa, N.M., Ima, N.S. & Ahmad Nazrun,
S. (2013). Labisia pumila regulates bone-related genes expressions in
postmenopausal ostereoporosis model. BMC Complementary &
Alternative Medicine: 13:217.
Fisher, R.F. & Binkley, D. (2000). Ecology and Management of Forest Soil. 3rd ed.
John Willey & sons, New York. Pp489.
Hyun-kyung ,C., Dong-hyun, K., Jin Wook, K., Sulaiman, N., Mohamad Roji, S., &
Chang S. P. (2010). Labisia pumila Extract Protects Skin Cells from
Photoaging Caused by Uvb Irradiation. Journal of Bioscience and
Bioengineering: 109(3), 291–296.
Intan, I.H. & Nik, H.N.H. (2015). Kenapa Kacip Fatimah? Dewan Bahasa dan
Pustaka, Kuala Lumpur.
Rozihawati, Z, Azmy, M. & Aminah, H. (2005). Pembiakan Tampang Melalui
Keratan Bahagian Batang Labisia pumila (kacip Fatimah).
Proceedings
of the Seminar on Medicinal and Aromatic Plants: 368─373.
Saari, A & Noraini, O. (2013). Strategic Planning, Issues, Prospects and the
Future of the Malaysian Herbal Industry. International Journal of
Academic Research in Accounting, Finance and Management Sciences:
91─102.
The Potash Development Association. 2011. Soil Analysis: Key to Nutrient
Management Planning. Available from: <www.pda.org.uk>. [26 January
2014].
152
TAPAK WARISAN FRIM: POTENSI SEBAGAI TAPAK KONSERVASI
EX
SITU TERBESAR BAGI GERMPLASMA TUMBUHAN UBATAN DI
SEMENANJUNG MALAYSIA
A Noorsiha, AM Ainnur Amira, M Kamariah, MK Mohd Haire, B Nuranis
Suraya & F Nur Fairuz
Tel: 03-6279 7618 / 03-6279 7619
ABSTRAK
Tapak Warisan FRIM merupakan hutan tropika regenerasi tertua di dunia,
merupakan bukti penting pembentukan hutan buatan manusia dari kawasan
gersang. Komitmen FRIM dalam pembangunan germplasma tumbuhan ubatan
dan peruntukan kawasan sebagai tapak konservasi ex situ, mampu mengangkat
tapak warisan ini sebagai pusat pengumpulan tumbuhan hidup utama negara.
Terdapat enam buah tapak konservasi ex situ yang menampung sejumlah 592
spesies, 335 genera dari 109 famili. Justeru, mengangkat Tapak Warisan FRIM
ini sebagai tapak konservasi ex situ berpotensi sebagai tapak koleksi
germplasma dan menyimpan komposisi spesies ubatan terbesar di
Semenanjung Malaysia.
Kata kunci: Tapak Warisan FRIM, germplasma, tumbuhan ubatan, tapak
konservasi ex situ
LATAR BELAKANG
Pengiktirafan FRIM Sebagai Tapak Warisan Kebangsaan
FRIM ditubuhkan dengan visi sebagai institusi penyelidikan yang terunggul dan
berautoritatif dalam penyelidikan perhutanan tropika di peringkat
antarabangsa. Kampus FRIM diwartakan sebagai Tapak Warisan Semula Jadi
Negara pada 2009 dan seterusnya pada 2012, diangkat sebagai Tapak Warisan
Kebangsaan. Kampus FRIM berkeluasan 544.31 ha merupakan satu-satunya
institusi penyelidikan di dunia diiktiraf sebagai tapak warisan. Justeru, pelbagai
usaha sedang dilaksanakan bagi menggerakkan FRIM sebagai Tapak Warisan
Dunia UNESCO menjelang tahun 2020. Tapak ini merupakan contoh terbaik
usaha sustainability science jangka panjang dan perlu diteruskan demi manfaat
sains sejagat serta bagi memastikan inovasi generasi terdahulu dapat dikongsi
dengan generasi akan datang.
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Pembentukan Hutan Tropika Regenerasi Tertua di Dunia
Hutan tropika regenerasi yang berusia hampir 100 tahun di Kampus FRIM,
terbentuk menerusi konsep “on-going ecological and biological processes”
merupakan bukti penting pembentukan hutan buatan manusia dari kawasan
gersang. Inovasi dan pengurusan yang dilaksanakan sejak tahun 1920-an
berjaya memulihkan kawasan yang dahulunya bekas lombong, kebun sayur dan
belukar, bertukar menjadi hutan regenerasi tertua dunia dan kini menjadi
model pemulihan hutan serta mendapat pengiktirafan pakar hutan tropika.
Kawasan hutan asal sebenarnya telah dibalak beberapa kali. Kini terdapat
pelbagai jenis dirian hutan yang dibangunkan menerusi penanaman satu
spesies atau pelbagai spesies merangkumi spesies tempatan dan eksotik
bernilai komersial.
Program Konservasi Ex Situ Spesies Hutan di Tapak Warisan FRIM
Komitmen FRIM untuk membangunkan tapak konservasi ex situ, akhirnya
mengangkat Kampus FRIM sebagai pusat pengumpulan tumbuhan hidup
utama negara. Pengutipan pelbagai flora spesies native dan eksotik
dilaksanakan menerusi berbagai ekspedisi botanikal di seluruh negara.
Pengumpulan secara aktif koleksi tumbuh-tumbuhan terancam bermula pada
tahun 2000 menerusi projek pembangunan germplasma spesies tumbuhan
langka dan terancam di Malaysia.
Pembentukan Tapak Konservasi Ex Situ Tumbuhan Ubatan
Taman Etnobotanikal
Taman Etnobotani ditubuhkan pada tahun 1995 sebagai pusat pemuliharaan ex
situ serta pusat domestikasi bagi tumbuhan ubatan dan beraroma yang
digunakan oleh golongan etnik di Semenanjung Malaysia. Berfungsi untuk
membangkitkan minat masyarakat umum dalam tumbuhan ubatan dan
beraroma, terdapat hampir 200 spesies tumbuhan mewakili 85 famili.
Taman Etnoflora, Taman Botani Kepong (KBG)
Pembangunan Taman Botani Kepong atau KBG dimulakan pada tahun 1996
dengan fungsi sebagai pusat kecemerlangan penyelidikan dan pendidikan
dalam bidang botani, hortikultur, landskap dan pendidikan alam sekitar.
Landskap KBG turut dicirikan dengan Taman Etnoflora yang kini mempamerkan
kira-kira 110 spesies tumbuhan ubatan, etnobotanikal dan beraroma.
154
Denai Alam FRIM
Sehingga tahun 2009, terdapat enam buah denai alam dibangunkan dengan
tema berbeza bagi pelawat berekreasi sambil mempelajari sumber hutan yang
ditemui di sepanjang perjalanan. Antara denai alam yang terawal menyimpan
pelbagai spesies tumbuhan ubatan ialah Denai Keruing, manakala Denai
Sebasah yang dibina pada tahun 2008 menggambarkan pula kemandirian flora
dan fauna di kawasan hutan becah.
Arboretum FRIM
Semenjak 1926 penyelidikan botani dan perhutanan memerlukan sokongan
koleksi tumbuhan hidup yang terurus. Kini, Arboretum FRIM telah menjadi
pusat pengumpulan tumbuhan hidup terbesar dan mempunyai koleksi yang
mewakili keseluruhan flora di Malaysia. Terdapat tujuh arboretum di Kampus
FRIM dan kini menyimpan kira-kira 354 spesies dari 149 genera mewakili 48
famili tumbuhan yang ditanam.
Rumah Tradisional Melayu Melaka
Berasal dari Kampong Padang Sebang, Alor Gajah, dibina pada tahun 1917 dan
kemudian dipindahkan ke Kampus FRIM pada 1988. Di perkarangan rumah ini
diserikan dengan pelbagai tanaman spesies tumbuhan ubatan bagi
menggambarkan budaya dan penghargaan masyarakat Melayu terhadap
tumbuhan yang berguna dalam kehidupan harian.
Plot Penyelidikan FRIM
Dianggarkan 380 ha merupakan kawasan hutan bertanam atau plot
penyelidikan dan dilaporkan menyimpan 2,434 spesies tumbuhan. Plot
penyelidikan yang mencapai usia hampir seratus tahun, kini mulai tersebar
dengan pelbagai spesies yang tidak ditanam tetapi tumbuh di bawah naungan
dirian hutan yang ditanam. Kewujudan pelbagai spesies tumbuhan ubatan di
plot penyelidikan memerlukan kajian lanjutan terutama aktiviti inventori
spesies.
Rajah 1. Peta taburan tapak-tapak konservasi ex situ tumbuhan ubatan di Tapak
Warisan FRIM
155
BAHAN DAN KAEDAH
Program Pembangunan Koleksi Germplasma Tumbuhan Ubatan
Antara program terawal bagi pembangunan koleksi germplasma tumbuhan
ubatan ialah menerusi Projek WWF-FRIM sekitar tahun 1990an melibatkan
pengutipan koleksi buah-buahan nadir, koleksi tumbuhan ubatan dan
beraroma di bawah Bahagian Tumbuhan Ubatan, program konservasi ex situ
koleksi buluh menerusi program IDRC, Projek Flora Malesiana Center (FMC-3)
iaitu Plant Introduction for Kepong Botanic Gardens termasuk program
pertukaran bahan tanaman dengan Taman Botani Pulau Pinang di bawah
projek (FMC-10)
Pembangunan Sistem Pangkalan Data Flora Kampus FRIM
Pembangunan sistem pangkalan data biodiversiti Kampus FRIM berjaya
mengkompilasikan pelbagai rekod tumbuhan yang pernah ditanam seawal
tahun 1923. Sehingga kini sejumlah 21,743 rekod telah terkumpul menerusi
pangkalan data myBOTANI berdasarkan 12 sumber maklumat botanikal
tersedia. Analisa rekod dan penyediaan senarai semak awal menggunapakai
PROSEA-Medicinal And Poisonous Plants Jilid 1-3 yang menyenaraikan sejumlah
1,300 spesies tumbuhan ubatan di kawasan Asia Tenggara. Ringkasan sumber
maklumat botanikal ditunjukkan pada Rajah 1.
Rajah 1. Tapak Warisan FRIM: bilangan rekod sumber maklumat botanikal
156
Berdasarkan 21,743 rekod dari pelombongan data (data mining) dan sistem
pangkalan data myBotani, dianggarkan terdapat 592 spesies mewakili 335
genera dari 109 famili tumbuhan ubatan di Kampus FRIM. Berdasarkan
anggaran awal keseluruhan 2,434 spesies tumbuhan yang ditemui, kira-kira
24.32% merupakan spesies ubatan. Manakala 10 famili utama tertera pada
Jadual 1.
Jadual 1. Sepuluh famili utama tumbuhan ubatan dan taburan di tapak
konservasi ex situ Kampus FRIM
*Tapak Konservasi Ex Situ
No
Famili
Genera
Spesies
1
2
3
4
5
6


5
10
Annonaceae
1





2
Euphorbiaceae
13
17




7
10
Lamiaceae
3
4
11




Lauraceae
4



18
26
Leguminosae
5
7
16




Moraceae
6
7
8
9
10
11
Myrtaceae
Palmae
Rubiaceae
Zingiberaceae
Pelbagai
Jumlah
8
6
13
9
245
335
17
11
19
12
443
592




















*Tapak konservasi ex situ: 1-Taman Etnobotani; 2-Taman Etnoflora Herba, KBG; 3-Rumah
Tradisional Melayu Melaka; 4-Denai Alam; 5-Arboretum; 6-Plot-Plot Penyelidikan FRIM
Secara keseluruhan, terdapat enam buah tapak konservasi ex situ utama bagi
koleksi germplasma tumbuhan ubatan. Didapati Taman Etnobotani FRIM
merupakan tapak ex situ yang menyimpan paling banyak koleksi germplasma
tumbuhan ubatan iaitu sejumlah 152 spesies, diikuti oleh plot-plot
penyelidikan FRIM (128 spesies) dan Taman Etnoflora, KBG (28 spesies) seperti
ditunjukkan di Rajah 2.
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Bilangan Spesies Tumbuhan Ubatan di Tapak Konservasi
Ex-Situ Kampus FRIM
10
Taman Etnobotani
Plot Penyelidikan
12
26
Taman Etnoflora KBG
Arboretum
28
Denai Alam
128
Rumah Tradisional
Melayu Melaka
152
0
50
100
150
200
Bilangan Koleksi ex-situ Spesies Tumbuhan Ubatan
Rajah 2. Bilangan spesies tumbuhan ubatan di enam tapak konservasi ex situ
Kampus FRIM
KESIMPULAN
Tapak Warisan FRIM merupakan bukti penting pembentukan hutan buatan
manusia dari kawasan gersang. Namun tapak warisan ini mampu bertindak
sebagai pusat pengumpulan tumbuhan hidup utama negara termasuk
pembangunan koleksi ex situ spesies ubatan. Terdapat enam buah tapak
konservasi ex situ yang menyimpan sejumlah 592 spesies, 335 genera dari 109
famili tumbuhan ubatan, sekali gus signifikan untuk mengangkat Tapak
Warisan FRIM sebagai warisan negara di persada dunia.
RUJUKAN
Padua, L.S., Bunyapraphatsara, N. & Lemmens, R. H. M. J. (Eds.), (1999). Plant
Resources of South-East Asia No 12(1). The Prosea Foundation,
Bogor, Indonesia. 705 pp.
Van Valkenburg, J. L. C. H. & Bunyapraphatsara, N. (Eds.). (2001). Plant
Resources of South-East Asia No 12(2). Medicinal and Poisonous
Plants. The Prosea Foundation. Backhuys Publishers, Leiden.776 pp.
Lemmens, R. H. M. J. & Bunyapraphatsara, N. (Eds.). (2003). Plant Resources of
South-East Asia No 12(2). Medicinal and Poisonous Plants. Prosea
Foundation, Backhuys Publishers, Leiden. 776 pp.
158
PENGGUNAAN TUMBUHAN DALAM KEHIDUPAN ORANG ASLI
KUMPULAN SENOI DI SEMENANJUNG MALAYSIA: KHUSUSNYA
SUBETNIK TEMIAR DAN SEMAI
AL Tan, A, BJ Chee, MN Madihah, M Mastura, B Intan Nurulhani, M Badariah,
R Rosniza, Z Nurul Husna, Y Norshakila, M Nik Musaadah & HF Lim
Tel: 03-6279 7671
Faks: 03-6272 9805
Dengan kerjasama komuniti Orang Asli
Jawatankuasa Pengetahuan Tradisi komuniti Temiar di RPS Banun, Gerik dan
Pos Tuel, Gua Musang, komuniti Semai di Ulu Geroh, Gopeng, komuniti Jahut di
Paya Mendoi, Kuala Krau, komuniti Semoq Beri di Sungai Berua, Kuala
Terengganu & komuniti Che Wong di Sungai Enggang, Lanchang
ABSTRAK
Populasi Orang Asli merangkumi 0.63% (178,197) penduduk yang menetap di
Semenanjung Malaysia. Orang Asli merupakan komuniti yang menetap di
pinggir kampung, pekan kecil, pesisiran sungai atau kawasan pedalaman di
mana kehidupan dan amalan hariannya amat dipengaruhi atau diwarisi
daripada nenek moyang. Orang Asli telah digolongkan kepada tiga kumpulan;
i.e. Negrito, Senoi dan Melayu Proto berdasarkan kepada budaya, bahasa,
kehidupan, kawasan rayau dan bentuk fizikal. Setiap kumpulan kemudian
dibahagikan kepada 18 subetnik dimana masing-masing merangkumi enam
subetnik bagi setiap kumpulan. Kumpulan Senoi merupakan kumpulan terbesar
yang terdiri daripada subetnik Semai, Temiar, Jahut, Semoq Beri, Mah Meri dan
Che Wong. Majoriti masyarakat kumpulan Senoi menetap di negeri Perak,
Pahang dan Kelantan kecuali komuniti subetnik Semoq Beri yang mendominasi
negeri Terangganu.
Maklumat berkenaan penggunaan tumbuhan dalam kehidupan Orang
Asli telah dikumpulkan berdasarkan lawatan ke beberapa Muzium Orang Asli,
pengumpulan data sekunder dan penerbitan berkaitan Orang Asli dan
pengamatan kehidupan harian masyarakat Orang Asli di beberapa kampung
semasa kerja lapangan. Masyarakat Orang Asli pada masa kini didapati masih
banyak menggunakan bahan berasaskan tumbuhan dalam kehidupan harian
terutama yang menetap di kawasan pedalaman. Beberapa aspek kehidupan
berasaskan penggunaan tumbuhan yang masih diamalkan oleh masyarakat
kumpulan Senoi adalah seperti makanan, peralatan memasak, kediaman,
pengangkutan (rakit & perahu), peralatan memburu (sumpit, tabung damak,
jerat binatang), menangkap ikan (bubu, serempang), kraftangan (bakul, tikar,
159
nyiru), perhiasan diri (tempok, rantai manik), alat muzik (tensol, genggong),
upacara sewang atau pemujaan roh (Hari Moyang, sakit, keraian), kematian,
kelahiran dan ukiran kayu (patung dan topeng). Masyarakat Orang Asli Senoi
didapati amat bergantung kepada spesies tumbuhan daripada jenis buluh,
palma (e.g. rotan, pinang, bertam, palas) pisang, mengkuang dan terap selain
daripada kutipan buah-buah dan pucuk tumbuhan dalam hutan.
Kata kunci: Orang Asli, Senoi, tumbuhan, kehidupan, subetnik
PENGENALAN
Berdasarkan kajian sejarah, masyarakat Senoi dipercayai berhijrah dari Indo
China ke Semenanjung Malaysia melalui Selatan Thailand kira-kira 500 tahun
dahulu. Masyarakat Senoi merupakan kumpulan terbesar (55%) daripada
populasi Orang Asli di Semenanjung Malaysia. Daripada 6 subetnik di bawah
kumpulan Senoi, subetnik Semai merupakan golongan yang terbesar (53%)
diikuti oleh Temiar (32%), Jahut, Semoq Beri, Mah Meri dan Che Wong. Secara
umum, taburan penempatan masyarakat Senoi lazimnya di bahagian tengah
Semenanjung Malaysia terutamanya di kawasan kaki banjaran gunung di Perak,
Kelantan, Pahang dan Selangor. Secara fizikalnya, masyarakat Senoi didapati
mempunyai sedikit pebezaan dengan kumpulan Negrito iaitu berbadan agak
tinggi, berkulit lebih cerah dengan rambut beralun dan tidak kerinting
manakala bahasa yang digunakan adalah Austro-Asiatik (Hood 2006).
Taburan penempatan komuniti Semai adalah tinggi di kawasan
banjaran Titiwangsa di negeri Perak (Perak Tengah dan Perak Selatan) dan
bahagian barat Pahang manakala komuniti Temiar pula lebih tertumpu di Perak
Utara dan Kelantan. Komuniti Semai mengamalkan pertanian sara diri dengan
menanam padi huma, jagung, ubi kayu, pisang, keladi dan sayur-sayuran. Kini,
komuniti ini didapati telah mengusahakan ladang getah, kelapa sawit, buahbuahan (durian, petai) atau digaji sebagai pekerja di sektor awam atau
persendirian. Berbanding dengan komuniti Semai, komuniti Temiar menyara
kehidupan dengan aktiviti bercucuk tanam, berburu dan mencari tumbuhtumbuhan hutan. Komuniti ini amat berhati-hati dalam memilih lokasi untuk
membuat penempatan kerana mengamalkan kepercayaan animisma dan juga
percaya terdapat hantu atau penunggu di sesuatu tempat. Maka, pemilihan
untuk membuat kediaman mereka perlulah jauh daripada tanah perkuburan,
bebas daripada kayu keras serta kawasan berpaya, air terjun dan sungai besar.
160
BAHAN DAN KAEDAH
Maklumat berkenaan latar belakang dan penggunaan tumbuhan dalam
kehidupan Orang Asli telah dikumpulkan berdasarkan lawatan ke empat
Muzium Orang Asli iaitu Muzium Orang Asli Gombak, Selangor, Muzium Orang
Asli Jeli, Kelantan, Muzium Orang Asli Ayer Keroh, Melaka, Muzium Seni Kraf
Orang Asli, Kuala Lumpur berserta Festival Kesenian Orang Asal Antarabangsa
Selangor 2015 di Taman Botani Negara Shah Alam. Di samping itu,
pengumpulan data sekunder dan penerbitan buku dan video berkaitan Orang
Asli dan pengamatan kehidupan harian masyarakat Orang Asli di beberapa
buah kampung semasa kerja lapangan turut dijalankan. Temu bual turut
dijalankan di kalangan tok batin semasa Festival Kesenian Orang Asal
Antarabangsa Selangor 2015, petugas di muzium dan juga beberapa penduduk
kampung di kawasan kajian.
Secara umumnya, masyarakat Orang Asli pada zaman dahulu amat bergantung
kepada bahan berasaskan tumbuhan di sekeliling kediaman mereka untuk
pakaian dan keperluan kehidupan harian. Pakaian mereka diperbuat daripada
kulit kayu terap di mana lelaki akan memakainya sebagai cawat manakala
perempuan pula akan memakainya sebagai skirt pendek dengan membiarkan
bahagian atas tubuhnya terdedah. Walau bagaimanapun, Orang Asli pada
zaman sekarang telah memakai pakaian seperti masyarakat moden. Pakaian
daripada kulit kayu terap hanya akan digunakan semasa upacara tertentu atau
ketika berburu. Dalam arus kemodenan, beberapa makanan tradisi yang masih
diamalkan oleh masyarakat Orang Asli adalah seperti nasi dalam buluh, ayam
dalam buluh, pulut dalam periuk kera, ubi bakar, ikan pais, ubi lempak, ubi
kacau dan ikan dalam buluh. Peralatan dapur yang digunakan pula banyak
dihasilkan daripada buluh (cawan, bekas air, bekas nasi/lauk, peniup api,
sudip), tempurung kelapa (sudu, sengkalam,senduk, bekas nasi/lauk), upih
pinang (pinggan, timba), rotan (nyiru,pemarut) dan bertam (pemarut).
Kediaman komuniti Semai lazimnya dibina menggunakan buluh dan
kayu yang dibelah sebagai lantai sementara dindingnya diperbuat daripada
anyaman pelepah bertam manakala atapnya juga merupakan jalinan rapi
daripada pelepah bertam atau serdang. Rotan lazimnya digunakan untuk
mengikat tiang, alang dan lantai. Antara jenis rumah Semai adalah Deg Yeau menyimpan padi, Deg ran -atas pokok, Deg pisang sesikat -pondok sementara
dan rumah biasa -corak mengikut kreativiti individu (JHEOA 2002). Rumah
kediaman komuniti Temiar pula menggunakan buluh (bulat-bulat/ anyaman)
dan kayu-kayu dari pokok hutan sebagai lantai dan dindingnya manakala
161
atapnya merupakan jalinan pelepah bertam atau nipah yang rapi. Antara jenis
rumah yang popular dibina oleh komuniti Temiar adalah seperti Dekreh pondok, Deklare -rumah dengan hiasan dinding motif corak kelawar dan bunga
kelarai, Dek Rendetek -rumah buluh, Dek Tagon dan Dek Ulau -rumah panjang
dan Dek Jalaq Juh - atas pokok (Nisra Nisran 1995).
Komuniti Temiar sangat aktif dalam kegiatan memburu dan
menangkap ikan. Penggunaan buluh yang mempunyai ruas yang panjang
menjadi pilihan untuk membuat sumpit dan tabung damak. Ukiran bermotif
gunung, sungai dan hutan akan dihiasi pada sumpit dan tabung damak. Lidi
damak biasanya dibuat daripada pelepah langkap, enau, bertam atau serdang
manakala gabusnya pula biasanya dibuat daripada mahang dan pulai. Racun
pokok ipoh biasanya dicelup pada hujung lidi damak ketika memburu. Selain
daripada sumpit, pelbagai jenis jerat daripada buluh dan kayu-kayu hutan juga
dihasilkan untuk menangkap tupai (jerat tupai), lontong, khinzir dan rusa
(perangkap serkap), landak, burung dan ayam hutan (jerat ayam/jerat libas).
Pelbagai jenis bubu daripada buluh dan rotan dihasilkan oleh komuniti
Temiar di mana penggunaan setiap jenis bubu adalah bergantung kepada lokasi
sungai di sekitar kediamannya seperti bubu panchok, bubu tempirai, bubu Che
O’, tangkul dan ce’ak. Peralatan seperti serempang buluh dan kail (bertam)
serta perahu dan rakit (buluh) turut digunakan dalam aktiviti menangkap ikan.
Sementelahan pula, komuniti Semai banyak melibatkan diri dalam aktiviti
bercucuk tanam. Antara peralatan yang digunakan adalah seperti beliung (kayu
dan rotan), tabung getah (buluh), parang (kayu dan anyaman rotan), raga galas
(rotan), lesung kaki, kepok padi (upih pinang), lesung tumbuk padi, keri, kuku
kambing, pisau raut dan nyiru.
Komuniti Temiar dan Semai turut terkenal dengan amalan sewang.
Amalan sewang dijalankan sewaktu upacara berubat orang sakit, perkahwinan,
kelahiran dan kematian. Dalam upacara ini, banyak persiapan yang perlu
dilakukan seperti hiasan dinding, lilitan pinggang, selimpar, selak bed daripada
anyaman pelepah daun kelapa atau nipah, tempok (pelepah kelapa, palas,
rumput, manik) dan juga penggunaan pelbagai daun dan bunga yang berbau
wangi. Hiasan corak muka turut dilakukan ketika upacara sewang. Upacara
sewang biasanya akan diiringi oleh muzik tradisional yang dimainkan
menggunakan peralatan seperti gooh (buluh centong), kreb, bateq (gendang),
gong, genggong dan pensol (buluh). Sewang untuk tujuan berubat hanya akan
dijalankan pada waktu malam dan dihadiri oleh keluarga yang terlibat sahaja.
Upacara sewang ini akan diketuai oleh Halak dengan diiringi lagu dan tarian
pemujaan yang panjang dan mengambil masa 3-7 malam sehingga pesakit
berjaya diubati.
162
Semasa upacara perkahwinan, bujam (pandan) akan digunakan untuk
meletak pinang, kapur, sireh, gambir, bunga cengkih dan rokok tembakau.
Perhiasan pengantin seperti rantai manik (biji keril- Temiar) atau rantai buluh
perindu (Semai), selimpar (nipah), simpai (rotan) dan tempok (rumput, nipah,
kelapa) serta lilitan pinggang (kelapa, nipah) turut digunakan. Dalam upacara
kelahiran, uri budak dari komuniti Semai akan disimpan di dalam bekas yang
dimasukkan tanah terlebih dahulu sebelum disembur dan dijampi
menggunakan batang bonglai dan dikambus dengan tanah sebelum ditutup
dan diletakkan di dapur agar sentiasa panas supaya bayi tidak diganggu oleh
makhluk halus. Sebaliknya, komuniti Temiar pula akan membungkus uri bayi
berserta pisau buluh yang digunakan semasa upacara kelahiran dengan kain
untuk diikat di dahan pokok besar.
RUMUSAN
Secara kesimpulannya, masyarakat Orang Asli kumpulan Senoi masih
menggunakan pelbagai jenis pokok hutan dalam kehidupan seharian.
Walaupun dalam kumpulan yang sama, komuniti subetnik Temiar dan Semai
turut mempunyai sedikit perbezaan dari segi kraftangan yang dihasilkan dan
adat yang diamalkan.
RUJUKAN
Hood, S. (2006). Peoples and Traditions. The Encyclopedia of Malaysia Vol. 12.
Editions Didier Millet, Singapore. Pp. 148.
JHEOA (Jabatan Hal Ehwal Orang Asli) (2002). Kehidupan, Budaya dan Pantang
Larang Orang Asli. Kuala Lumpur: Perniagaan Rita. Pp. 154.
Nisra Nisran, A. R. (1995). Masyarakat Temiar di Gua Musang, Kelantan. Universiti
Pertanian Malaysia, Selangor.
163
PENGGUNAAN HERBA DALAM PERBIDANAN TRADISIONAL MELAYU
SELEPAS BERSALIN DI ZON TENGAH, SEMENANJUNG MALAYSIA
A Siti Khairul Bariyyah1,2
Institut Penyelidikan Perubatan, Pusat Penyelidikan Perubatan Herba, Jalan
Pahang, 50588, Kuala Lumpur
2
Universiti Tun Hussein Onn, Parit Raja, Batu Pahat, Johor
Tel: 03-2616 2666
Faks: 03-2693 9335
1
ABSTRAK
Herba merupakan komponen penting dalam perbidanan Melayu. Herba telah
digunakan oleh bidan kampung bersama amalan-amalan tertentu. Herba
digunakan bertujuan untuk membantu pemulihan dan pencegahan daripada
sebarang masalah kesihatan pada masa akan datang. Kajian ini merekodkan
dan mengenalpasti herba yang paling banyak digunakan sebagai ramuan oleh
bidan kampung dalam amalan-amalan tertentu dalam perbidanan Melayu
selepas bersalin. Bidan kampung dipilih dari senarai bidan yang dikumpulkan
oleh Institut Penyelidikan Perubatan (IMR) melalui projek ‘Study on Traditional
Medicine: midwives’, 2006–2007. Bidan yang dipilih berada dalam lingkungan
100 km dari Hospital Putrajaya, melibatkan negeri Selangor, Melaka, dan
Negeri Sembilan. Temu bual telah dijalankan dengan menggunakan borang kaji
selidik bagi mengumpul maklumat mengenai jenis herba yang digunakan serta
kaedah penghasilan ramuan tersebut. Seramai 30 orang bidan telah ditemu
bual, dan herba-herba yang dinyatakan direkodkan dan dibuat pengumpulan.
Kesemua responden didapati menggunakan herba dalam amalan mereka.
Sebanyak 14 amalan yang menggunakan herba direkod dan dianalisis.
Sebanyak 10 spesies tumbuhan yang paling banyak digunakan ialah kunyit,
lengkuas, halia, serai wangi, mengkudu, kayu manis, pandan wangi, limau nipis,
manjakani dan cekur. Kajian ini mendapati herba-herba yang lazim digunakan
dalam perbidanan Melayu adalah diketahui umum, mudah diperolehi dan juga
merupakan kegunaan harian masyarakat Melayu. Oleh itu, pendokumentasian
ini dapat memelihara ilmu perbidanan Melayu khususnya dalam penggunaan
herba; menyedarkan keperluan memelihara dan memulihara biodiversiti
negara yang kaya dengan herba serta menjadi takungan ilmu bagi rujukan pada
masa akan datang.
Kata kunci: herba, perbidanan, Melayu, tumbuhan ubatan
164
PENGENALAN
Malaysia merupakan salah satu negara megabiodiversiti yang kaya dengan
spesies flora dan fauna. Kekayaan sumber alami ini telah membekalkan sumber
ubatan tradisional kepada bangsa Melayu dan berbagai suku kaum bumiputera
di Malaysia (Samuel et al. 2010, Lin 2005, Ahmad & Ismail 2003, Ong Zuki &
Milow 2011, Zal et al. 2014). Penggunaan herba merupakan komponen penting
dalam perbidanan Melayu selepas bersalin. Herba digunakan dalam pelbagai
amalan dalam perubatan ini seperti ramuan air mandian ibu, ramuan sapuan
dan minuman tambahan. Walau bagaimanapun, melihat kepada kesan
pembangunan yang memberi impak kepada sosiobudaya tempatan, amalanamalan ini dikhuatiri semakin dipinggirkan (Ibrahim 2006).
BAHAN DAN KAEDAH KAJIAN
Kajian ini menggunakan kaedah temu bual ke atas 30 orang bidan kampung.
Kawasan kajian pula meliputi kawasan dalam lingkungan 100 km daripada
Hospital Putrajaya. Senarai bidan ini dipilih daripada data kajian Study on
Traditional medicine: Midwives yang dijalankan oleh Pusat Penyelidikan
Perubatan Herba pada tahun 2006 hingga 2007 di Semenanjung Malaysia.
Calon responden dihubungi melalui telefon untuk membuat temu janji.
Responden akan menandatangani borang keizinan) (PIC) dan perkongsian
faedah (ABS) sebelum ditemu bual. Temu bual dilakukan dengan
berpandukan soalan kaji selidik yang telah dibina. Maklumat dicatat dan data
terkumpul dianalisis.
Sebanyak 80 spesies tumbuhan telah dilaporkan dan hampir kesemua amalan
menggunakan campuran pelbagai herba dalam ramuan (Jadual 1).
Penggunaan herba dalam perbidanan tradisional Melayu selepas
bersalin juga digunapakai secara luaran (param, pilis, balut tungku, minyak
urut, tapel, air cebok) dan dalaman (jamu, ubat periuk, minuman herba).
Justeru, penggunaan herba melengkapkan amalan-amalan fizikal dalam
perbidanan Melayu selepas bersalin dan memberi kesan positif kepada
kesihatan dan kesejahteraan ibu.
165
Jadual 1. Senarai spesies tumbuhan dan kegunaannya perbidanan tradisional
Melayu
Bil.
Ramuan param, tangas kering dan mandi daun.
2.
Nama
tempatan
Asam gelugor
/keping
Asam jawa
3.
Akar angin
Ramuan param
4.
Betik jantan
Balut tungku (daun)
5.
Buah keras
Ramuan mandi daun
6.
Bunga lawang
Ramuan ubat periuk, jamu
7.
Bawang putih
Ramuan tangas kering (kulit), tangas basah (kulit),
ubat periuk (rizom), minyak urut (rizom)
8.
Bawang merah
Ramuan tangas kering (kulit), tangas basah (kulit)
9.
Bonglai
Ramuan mandi daun (daun), balut tungku (daun),
minyak urut
10.
Balik angin
Ramuan mandi daun (daun)
11.
Cermai
12.
Cengkih
Ramuan param, pilis, ubat periuk, jamu, minyak urut
13.
Cabai pintal
Ramuan pilis, ubat periuk, jamu
14.
Cucur atap
Ramuan pilis
15.
Cina maki
16.
Chemumar
17.
Cekur
18.
Dukung anak
Ramuan param, ubat periuk, jamu (rizom), mandi
daun (daun, rizom), minyak urut
19.
Delima
Ramuan ubat periuk
20.
Ramuan minyak urut
22.
Ginseng
(tempatan)
Hempedu bumi
/Pokok cerita
Halia
23.
Halba
24.
Halia bara/ halia
merah
Ramuan param, tangas kering, tangas basah, mandi
daun
Ramuan param, ubat periuk, jamu, mandi daun
(daun), minyak urut
1.
21.
Penggunaan tradisional
Ramuan pilis, ubat periuk, jamu, mandi daun.
Ramuan ubat periuk
Ramuan param (rizom), ubat periuk, tangas basah,
jamu, mandi daun, isi tungku (rizom), minyak urut
166
25.
26.
Inai
Jemuju
Ramuan ubat periuk, jamu, mandi daun (daun)
Ramuan ubat periuk,
27.
Jintan manis
Ramuan pilis, ubat periuk, jamu, tangas basah, mandi
daun, isi tungku
28.
Jintan putih
daun, isi tungku
29.
Jerangau
30.
Jarak pagar
Ramuan param, tangas basah (daun), mandi daun
(daun), minyak urut
Balut tungku (daun)
31.
Jering (kering)
Ramuan jamu
32.
Jambu batu
33.
Jarum emas
Ramuan tangas basah (daun, pucuk), mandi daun
(daun)
Ramuan ubat periuk
34.
Jintan hitam
Ramuan pilis, jamu, tangas basah, isi tungku, minyak
urut
35.
Ketumbar
Ramuan ubat periuk, jamu, tangas basah, mandi daun
36.
Kayu manis
Ramuan param, pilis, ubat periuk, jamu, minyak urut
37.
Kancing baju
Ramuan ubat periuk
38.
Kacip fatimah
Ramuan ubat periuk, Ramuan jamu (bunga)
39.
Kayu putih
Ramuan ubat periuk
40.
Kaduk
41.
Kelapa
Ramuan minyak urut (minyak)
42.
Kunyit
43.
Sekentut
Ramua param, pilis, ubat periuk, jamu (daun, rizom),
tangas basah (daun), mandi daun (daun), balut
tungku (daun), minyak urut
Ramuan param
44.
Kantan
Balut tungku (daun)
45.
46.
Kayu serapat
Kayu angin
Ramuan ubat periuk, jamu
Ramuan ubat periuk, jamu, isi tungku
47.
Lalang
48.
Lada hitam
Ramuan param (buah), ubat periuk (buah), jamu
(buah), tangas basah (buah)
49.
Limau purut
Ramuan param (jus buah), tangas basah (daun,
batang), mandi daun (daun, batang)
50.
Limau nipis
51.
Lemuni
Ramuan param (jus buah), pilis, tangas basah, mandi
daun (daun)
52.
Lemuni hitam
167
53.
Lengkuas
Ramuan jamu, tangas kering, tangas basah (daun),
mandi daun (daun), balut tungku (daun), isi tungku
(rizom), minyak urut
Ramuan jamu, tangas basah, mandi daun, minyak
urut
Ramuan param, ubat periuk, jamu, tangas basah, isi
tungku
Ramuan tangas kering, mandi daun (daun), balut
tungku (daun)
54.
Lempuyang
55.
56.
57.
Lada ekor
Lada sireh
Manjakani
58.
Mengkudu
59.
Nilam
Ramuan mandi daun
60.
Pinang
Ramuan ubat periuk
61.
62.
Padi (Beras)
Pala
Ramuan param (buah), pilis (buah)
Ramuan param
63.
Putarwali
Ramuan ubat periuk
64.
Pandan wangi
65.
66.
Pegaga
Pelaga
(daun), balut tungku (daun)
Ramuan pilis, ubat periuk, mandi daun
Ramuan jamu (buah)
67.
Rambai
Balut tungku (daun)
68.
Rambutan
69.
Serai wangi
(daun), minyak urut, balut tungku (daun)
70.
Seringan
71.
Senduduk
Ramuan ubat periuk
72.
Sireh
daun (daun, batang), balut tungku (daun)
73.
Serai
74.
Sembong
75.
Tetulang
Ramuan mandi daun (daun, batang)
Balut tungku (daun)
Ramuan tangas basah (daun)
Ramuan tangas basah (pokok), mandi daun (pokok)
76.
Tengkuk biawak
Ramuan ubat periuk
77.
Turi
Ramuan pilis
78.
79.
80.
Temu lawak
Temu kunci
Temu ireng
Ramuan ubat periuk, jamu, mandi daun
Ramuan ubat periuk, jamu, mandi daun
Ramuan ubat periuk
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RUMUSAN
Penggunaan herba merupakan komponen penting dalam perbidanan
tradisional Melayu selepas bersalin dan masih lagi diamalkan sehingga kini.
Melalui pendokumentasian ini, nilai intrinsik pengetahuan tradisional termasuk
nilai intelek, pendidikan, sosial, ekonomi dan saintifik dapat diiktiraf dan
diperakui bahawa sistem pengetahuan tradisional merupakan kerangka inovasi
berterusan yang berfaedah kepada manusia. Oleh itu, pengetahuan tradisional
perlu dipelihara, didokumentasi dan diteruskan agar tidak pupus ditelan arus
pembangunan.
PENGHARGAAN
Setinggi penghargaan diucapkan kepada responden kajian ini yang telah
memberikan kerjasama dalam kajian ini.
RUJUKAN
Ahmad, F. B. & Ismail, G. (2003). Medicinal Plants Used by Kadazandusun
Communities around Crocker Range. ASEAN Review of Biodiversity and
Environmental Conservation (ARBEC).
Ibrahim, Y. (2006). Komuniti Melayu Pulau Tioman: Isu dan Cabaran dalam
Konteks Pembangunan Pelancongan. Jurnal Melayu, 2: 141–153.
Lin, K. W. (2005). Ethnobotanical Study of Medicinal Plants Used by The Jah Hut
Peoples in Malaysia. Indian Journal of Medical Sciences, 59(4): 156.
Ong, H. C., Zuki, R. M. & Milow, P. (2011). Traditional Knowledge of Medicinal
Plants Among The Malay Villagers in Kampung Kemas, Terengganu.
Malaysia. Ethno Med, 5(3): 175 –185.
Samuel, A. J. S. J. et al. (2010). Ethnomedical Survey of Plants Used by The
Orang Asli in Kampung Bawong, Perak, West Malaysia. Journal of
Ethnobiology and Ethnomedicine, 6(5): 1–6.
Zal, W. A., Salleh, H., Omar, M., & Halim, S. (2014). Kebolehcapaian dan
Keterancaman Modal Semula Jadi Orang Asli Lanoh di
Malaysia. Geografia: Malaysian Journal of Society and Space, 10(2): 178–
188.
169
TREND PENGGUNAAN 18 SPESIES TUMBUHAN UBATAN DI BAWAH
PROGRAM NKEA DI KALANGAN PENGAMAL PERUBATAN TRADISIONAL
MELAYU DI SEMENANJUNG MALAYSIA
M Dionysia, MS Abdul Hayat, M Nik Musaadah, B Intan Nurulhani, MN
Madihah, Z Nurul Husna, J Fadzureena, HF Lim, AL Tan, R Rosniza, MA Nor
Azah, M Mastura & H Norini
Tel: 03-6279 7683
ABSTRAK
Di bawah program National Key Economic Area (NKEA) Herba EPP#1, sebanyak
18 spesies tumbuhan ubatan telah dikenal pasti sebagai spesies yang
mempunyai potensi ekonomi. Justeru, pendekatan atau pengenalan spesies
tersebut kepada orang ramai adalah perlu untuk memartabatkan lagi
kepentingan dan penggunaan spesies tumbuhan ubatan tersebut dalam
menyumbang kepada peningkatan ekonomi negara. Analisis penggunaan
spesies tumbuhan ubatan di bawah program NKEA oleh 355 orang pengamal
perubatan tradisional Melayu di seluruh Semenanjung Malaysia telah
dilaksanakan hasil daripada pelaksanaan survei dalam
projek
Pendokumentasian Pengetahuan Tradisi Melayu Berkaitan Tumbuhan Ubatan
di Semenanjung Malaysia. Empat spesies NKEA yang paling popular digunakan
oleh pengamal ialah mengkudu (33%), dukung anak (29%), halia (26%) dan
senduduk (17%) manakala spesies yang paling sedikit digunakan adalah kelo
(1%) dan rosel (0.3%). Amalan perubatan terutama sekali yang melibatkan
penggunaan tumbuhan dapat menjadi asas kepercayaan atau keyakinan orang
ramai untuk menggunakan tumbuhan tersebut sebagai sumber perubatan,
seterusnya menyumbang kepada usaha untuk memperkenalkan spesies
tumbuhan ubatan yang mempunyai nilai potensi ekonomi kepada masyarakat.
Kata kunci: pengetahuan tradisi Melayu, tumbuhan ubatan, NKEA
170
PENGENALAN
Selaras dengan dasar kerajaan untuk menyumbang kepada peningkatan tahap
ekonomi negara, 18 spesies tumbuhan ubatan telah diberi keutamaan di
bawah program NKEA Pertanian dalam NKEA Herba EPP#1. Antaranya ialah
tongkat ali (Eurycoma longifolia), kacip fatimah (Labisia pumila), hempedu
bumi (Andrographis paniculata), misai kucing (Orthosiphon stamineus),
mengkudu (Morinda citrifolia), dukung anak (Phyllanthus amarus), halia
(Zingiber officinale), mas cotek (Ficus deltoidea), rosel (Hibiscus sabdariffa),
pegaga (Centella asiatica), belalai gajah (Clinacanthus nutans), gelenggang
(Senna alata), lempoyang (Zingiber zerumbet), kaduk (Piper sarmentosum),
peria katak (Momordica charantia), kelo (Moringa oleifera), senduduk
(Melastoma malabathricum) dan sambung nyawa (Gynura procumbens).
Namun sejauh manakah pendekatan atau tahap pengetahuan masyarakat
Melayu tentang penggunaan spesies tersebut dalam amalan perubatan
tradisional? Dalam konteks amalan perubatan tradisional terutamanya yang
menggunakan tumbuhan ubatan dalam rawatan, amalan ini dapat
menyumbang kepada usaha memperkenalkan spesies tumbuhan ubatan yang
mempunyai potensi ekonomi berdasarkan kajian saintifik yang telah
dilaksanakan dan membuktikan tumbuhan tersebut mempunyai nilai
perubatan.
Amalan atau pengetahuan tradisi juga merupakan salah satu faktor
yang boleh menyumbang kepada penemuan spesies yang berpotensi untuk
dikomersialkan. Kekerapan penggunaan spesies tumbuhan ubatan dalam
perubatan boleh dijadikan sebagai testimoni atau bukti kemujaraban
tumbuhan tersebut, seterusnya memperlihatkan potensi tumbuhan tersebut
untuk dikomersialkan.
Justeru, kajian ini telah menganalisa trend penggunaan spesies yang
tersenarai di bawah program NKEA di kalangan 355 orang pengamal perubatan
tradisional Melayu di Semenanjung Malaysia dan seterusnya mengenalpasti
spesies tumbuhan ubatan yang paling popular digunakan dan penggunaannya
dalam amalan perubatan tradisional.
KAEDAH
Temu bual secara terperinci mengenai penggunaaan tumbuhan ubatan telah
dijalankan ke atas 355 orang pengamal perubatan Melayu di Semenanjung
Malaysia. Selain daripada maklumat hasil temu bual, kutipan sampel tumbuhan
yang digunakan oleh para pengamal dalam perubatan turut dilakukan bagi
tujuan pengecaman nama spesies. Seterusnya, analisis terhadap penggunaan
171
tumbuhan di kalangan pengamal dilaksanakan bagi mendapatkan maklumat
lebih terperinci tentang trend pengunaan serta tujuan penggunaan tumbuhan
berkenaan.
Berdasarkan survei yang dijalankan ke atas 355 orang pengamal perubatan
tradisional Melayu, didapati bahawa pokok mengkudu, dukung anak, halia dan
senduduk antara tumbuhan popular yang digunakan oleh para pengamal
berbanding kelo dan rosel. Peratus penggunaan dan jumlah pengamal yang
menggunakan tumbuhan ubatan di bawah program NKEA adalah seperti di
Jadual 1.
Kegunaan spesies tumbuhan dalam perubatan tradisional dan ciri-ciri
tumbuhan merupakan antara faktor yang mempengaruhi trend penggunaan
tumbuhan tersebut. Secara tradisinya, daun mengkudu biasanya digunakan
sebagai lapik tungku dan juga dimakan sebagai ulam. Pokok dukung anak
biasanya dijadikan sebagai ubat demam kuning atau demam panas kanakkanak. Halia pula selalunya dijadikan sebagai bahan untuk membuat minyak
urut bagi membuang angin dalam badan dan juga sebagai bahan perisa dalam
masakan. Daun senduduk digunakan untuk mengubati luka kecil dan buahnya
juga boleh dimakan. Kegunaan tumbuhan tersebut sangat berkait rapat kepada
konsep perubatan tradisional yang selalunya memberi fokus kepada perawatan
penyakit-penyakit ringan. Dalam kajian ini juga terdapat beberapa spesies
tumbuhan ubatan selain spesies tumbuhan di bawah program NKEA yang turut
popular digunakan dalam perubatan tradisional Melayu seperti serai wangi,
kunyit, pandan, inai, sireh, pisang kelat, seringan, jerangau, lada hitam dan
capa/sembung. Spesies-spesies tumbuhan tersebut kerap digunakan dalam
rawatan atau penjagaan kesihatan wanita selepas bersalin seperti mandian
selepas bersalin, lapik tungku, dan jamu/maajun selepas bersalin. Jerangau
pula selalu dikaitkan dengan ilmu mistik untuk menghalau makhluk-makluk
halus yang mengganggu terutama bayi.
Kekerapan penggunaan spesies tumbuhan yang senang dijumpai atau
mudah diperoleh di kawasan persekitaran lebih tinggi daripada spesies
tumbuhan yang sukar diperoleh. Contohnya, penggunaan pokok mengkudu
dan dukung anak yang selalunya tumbuh liar di kawasan rumah atau kawasan
pinggir kampung lebih tinggi berbanding tongkat ali atau kacip fatimah yang
hanya dapat dijumpai di kawasan hutan.
Spesies tumbuhan di bawah program NKEA yang paling kurang
digunakan oleh pengamal ialah kelo dan rosel yang mana dipercayai bukan
172
tumbuhan asal di Malaysia. Kelo dipercayai berasal dari India dan rosel juga
dikatakan berasal dari India dan Afrika Barat. Tumbuhan yang diperkenalkan
dari negara luar tersebut mungkin merupakan salah satu faktor ianya tidak
kerap digunakan dan kelihatan sangat asing di kalangan pengamal perubatan
tradisional Melayu.
Jadual 1. Penggunaan tumbuhan ubatan di bawah program NKEA oleh
pengamal perubatan tradisional Melayu (n=355)
Bil.
Nama
tempatan
herba
Famili
Nama Saintifik
Bil
pengamal
%
penggunaan
1.
2.
Mengkudu
Dukung anak
Rubiaceae
Phyllanthaceae
117
101
33
29
3.
4.
Halia
Senduduk
Zingiberaceae
Melastomataceae
93
60
26
17
5.
6.
Pegaga
Tongkat ali
Apiaceae
Simaroubaceae
51
50
14
14
7.
Acanthaceae
47
13
8.
9.
Hempedu
bumi
Kacip fatimah
Misai kucing
44
43
12
12
10.
11.
12.
Gelenggang
Lempoyang
Belalai gajah
Fabaceae
Zingiberaceae
Acanthaceae
39
31
25
11
9
7
13.
Kaduk
Piperaceae
25
7
14.
15.
Moraceae
Asteraceae
15
15
4
4
16.
Mas cotek
Sambung
nyawa
Peria katak
13
4
17.
18.
Kelo
Rosel
Moringaceae
Malvaceae
Morinda citrifolia
Phyllanthus
amarus
Zingiber officinale
Melastoma
malabathricum
Centella asiatica
Eurycoma
longifolia
Andrographis
paniculata
Labisia pumila
Orthosiphon
stamineus
Senna alata
Zingiber zerumbet
Clinacanthus
nutans
Piper
sarmentosum
Ficus deltoidea
Gynura
procumbens
Momordica
charantia
Moringa oleifera
Hibiscus sabdariffa
2
1
1
0.3
Myrsinaceae
Lamiaceae
Cucurbitaceae
KESIMPULAN
Penggunaan tumbuhan ubatan dalam pengetahuan tradisi menjadi indikator
penting dalam menyumbang ke arah peningkatan potensi herba untuk
dikomersialkan dan seterusnya membantu mengetengahkan populariti spesies
tumbuhan ubatan di kalangan masyarakat. Penemuan spesies tumbuhan
ubatan yang banyak digunakan dalam perawatan pelbagai penyakit
173
membolehkan para penyelidik melakukan kajian saintifik untuk membuktikan
kesahihan tumbuhan ubatan tersebut dijadikan ubat kepada penyakit tertentu
tanpa memberi kesan mudarat kepada pengguna. Pengetahuan tradisi juga
turut menjadi asas penting untuk memartabatkan spesies komersial
berdasarkan kepentingan dan penggunaannya dalam masyarakat. Namun
demikian, pendekatan yang lebih meluas tentang penggunaan spesies
tumbuhan terutama tumbuhan yang mempunyai potensi ekonomi adalah perlu
supaya populariti penggunaan tumbuhan lebih tinggi seterusnya menyumbang
kepada permintaan yang tinggi terhadap produk herba dalam pasaran.
PENGHARGAAN
Penulis mengucapkan terima kasih kepada semua pengamal perubatan
tradisional Melayu dan pegawai pengumpul data yang terlibat dalam
menjalankan survei dan juga pihak Kementerian Pertanian dan Industri Asas
Tani (MOA) yang membiayai dana penyelidikan.
RUJUKAN
http://www.worldagroforestry.org/treedb/AFTPDFS/Hibiscus_sabdariffa.PDF
http://www.worldagroforestry.org/treedb/AFTPDFS/Moringa_oleifera.PDF
Kementerian Industri Asas Tani. Panduan Pengendalian Borang Skim Geran
Penyelidikan NKEA HERBA EPP#1 (NRGS). www.moa.gov.my.
174
AMALAN PERUBATAN TRADISIONAL MELAYU DI SEMENANJUNG
MALAYSIA: ADAKAH MASIH RELEVAN PADA MASA KINI ?
MS Abdul Hayat, M Dionysia, M Nik Musaadah, B Intan Nurulhani, Z Nurul
Husna, J Fadzureena, R Rosniza, MN Madihah, AL Tan, HF Lim, MA Nor Azah,
MS Md. Azharulzaman, S Fatin Nadiah, M Mastura & H Norini
Tel: 03-6279 7683
ABSTRAK
Amalan perubatan tradisional Melayu merupakan ilmu yang diwarisi sejak
bergenerasi dalam mengekalkan kesihatan, merawat dan menyembuhkan
pelbagai penyakit, merangkumi bidang perbidanan, urutan, jampi dan bekam.
Justeru, bancian dan survei bagi mendapatkan maklumat pengamal perubatan
tradisional Melayu (PTM) telah dilaksanakan di Semenanjung Malaysia.
Berdasarkan bancian dan survei yang telah dijalankan pada tahun 2013-2014,
sejumlah 4,968 individu telah dikenal pasti dan hanya 2,935 individu ditemu
bual. Daripada jumlah tersebut, sejumlah 355 orang pengamal telah dipilih dan
disurvei bagi mendapatkan maklumat yang lebih terperinci dalam penggunaan
tumbuhan ubatan semasa pengamalan. Amalan perubatan Melayu dibahagikan
kepada empat kategori iaitu perubatan tradisional Melayu khusus, umum,
perbidanan dan campuran. Sosioekonomi dan penempatan pengamal
perubatan tradisonal Melayu di luar bandar dan bandar menjadi salah satu
fokus di dalam kajian ini.
Kata kunci: pengamal perubatan tradisional Melayu, perbidanan, urutan, profil
pengamal, pengkhususan pengamalan
PENGENALAN
Perubatan tradisonal masyarakat Melayu yang diwarisi secara turun temurun
merupakan salah satu alternatif bagi merawat penyakit pada masa kini.
Perubatan tradisional masyarakat Melayu sering dikaitkan dengan penggunaan
tumbuhan ubatan semasa rawatan. Pengetahuan penggunaan tumbuhan
ubatan ini perlu didokumentasi bagi mengekalkan kesinambungan ilmu
pengetahuan perubatan tradisional masyarakat Melayu. Kajian ini
menumpukan profil sosio-ekonomi serta kesesuaian khidmat pengamal
perubatan traditional Melayu (PTM) pada masa kini.
175
KAEDAH
i.
ii.
iii.
Bancian dilakukan dengan mengenal pasti pengamal PTM melalui
pengerusi
jawatankuasa
kemajuan
dan
keselamatan
kampung/penghulu/penggawa/pengerusi persatuan penduduk.
Hasil daripada bancian, 355 pengamal PTM telah dipilih untuk
menyertai aktiviti survei. Temu bual separa berstruktur secara
bersemuka telah dilaksanakan.
Maklumat sosio-demografik dan maklumat kegunaan tumbuhan
dicatat dan direkodkan.
1. Profil Pengamal
Daripada 355 pengamal yang ditemu ramah, seramai 229 (64.5%) adalah
perempuan manakala selebihnya, iaitu 126 (35.5%) adalah lelaki. Ini
menunjukkan bahawa penglibatan dalam perubatan tradisional Melayu
dipelopori oleh kaum wanita. Keadaan ini boleh dikaitkan dengan permintaan
terhadap perkhidmatan perbidanan untuk tujuan kesihatan dan kecantikan di
kalangan wanita. Dari segi umur, 62% iaitu seramai 219 individu PTM berumur
lebih daripada 60 tahun, manakala 38% individu lagi (136 orang) terdiri
daripada golongan dalam lingkungan umur 40-60 tahun. Pengamal PTM yang
masih aktif adalah dalam lingkungan umur 51-80 tahun, iaitu melibatkan 321
orang (90%) daripada jumlah keseluruhan pengamal PTM.
2. Taburan Lokasi Pengamal Yang Ditemubual
Hasil kajian menunjukkan kebanyakan pengamal yang ditemuramah adalah
dari negeri Kedah iaitu 25%, diikuti oleh negeri Pahang sebanyak 17%, dan 12%
dari negeri Kelantan. Seramai lima orang pengamal sahaja (yang memenuhi
kriteria) berasal dari Melaka. Hasil kajian mendapati kebanyakan pengamal
PTM berada di kawasan luar bandar iaitu sebanyak 227 individu (64%)
manakala 128 individu (36%) berada di kawasan bandar. Ini memberi
gambaran bahawa permintaan terhadap pengamalan PTM masih lagi tertumpu
di kawasan luar bandar. Daripada pecahan lokasi tersebut, 64% pengamal PTM
yang berada di luar bandar adalah pengamal perubatan umum (28.5% atau 101
individu), 11.5% pengamal perbidanan (41 individu), 12.2% pengamal khusus
(43 individu) dan 11.8% pengamal campuran (42 individu). Daripada 128
pengamal PTM yang tinggal di kawasan bandar pula, bidang pengamalan yang
paling banyak diceburi adalah pengamalan perubatan umum iaitu sebanyak
18.3% (65 individu), diikuti dengan bidang perubatan secara khusus sebanyak
176
7.3% (26 individu), campuran sebanyak 5.6% (20 individu) dan yang paling
sedikit ialah bidang perbidanan iaitu 4.8% (17 individu).
3. Pengkhususan Pengamal
Amalan perubatan tradisional Melayu dikategorikan kepada empat kumpulan
iaitu perubatan umum, perbidanan, khusus dan campuran. Perubatan umum
merupakan bidang pengkhususan yang paling banyak diamalkan iaitu sebanyak
47% dan yang paling sedikit diceburi adalah bidang perbidanan (Rajah 1).
Rajah 1. Pengkhususan pengamal perubatan Melayu (n=355)
4. Produk Pengamal
Seramai 241 individu (daripada 355 orang pengamal) menghasilkan produk
menggunakan tumbuhan ubatan, manakala 114 individu lagi tidak
menghasilkan produk (Rajah 2). Produk yang sering dihasilkan adalah seperti
jamu/majun, minyak urut, kapsul kesihatan, ubat periuk dan air penawar.
Rajah 2. Jumlah pengamal yang menghasilkan produk
5. Jumlah Pengamal Mengikut Kelas Bilangan Pesakit
Hasil daripada survei yang dilaksanakan, didapati masih terdapat permintaan
untuk PTM (Jadual 1). Bilangan pesakit yang dirawat oleh 68% pengamal PTM
177
adalah dalam lingkungan 1-50 individu sebulan. Seramai 13 orang pengamal
PTM (4%) mampu merawat lebih daripada 200 pesakit dalam sebulan.
Jadual 1. Jumlah pengamal mengikut kelas bilangan pesakit sebulan
Bilangan pesakit
Jumlah pengamal
Peratus (%)
0-50
241
68
51-100
62
17
101-150
33
9
151-200
6
2
>200
13
4
Jumlah
355
100
6. Pedapatan Para Pengamal Berdasarkan Pengkhususan
Pendapatan individu bagi setiap pengamal perubatan menunjukan taraf
ekonomi mereka. Kebanyakan pengamal yang ditemu ramah mempunyai
sumber ekonomi tersendiri sama ada pendapatan berasaskan khidmat rawatan
ataupun pendapatan tetap daripada pekerjaan lain (Jadual 2). Sebilangan
pengamal PTM menjadikan pendapatan perkhidmatan rawatan sebagai
sumber ekonomi sampingan. Dari segi upah yang diterima, survei mendapati
hanya 289 responden (81%) menerima upah hasil daripada perkhidmatan yang
ditawarkan kepada para pesakit. Selebihnya, iaitu 66 orang pengamal (19%)
tidak menerima upah kerana melakukan perkhidmatan secara sukarela untuk
tujuan amal dan tidak mengenakan sebarang cas bayaran bagi perkhidmatan
yang ditawarkan.
Pengamal PTM yang dikelaskan sebagai perubatan umum merupakan
antara penerima pendapatan tertinggi hasil daripada amalan perubatan yang
dijalankan berbanding pengamal lain dengan jumlah RM 112,183.40 dan
purata pendapatan berjumlah RM 796.00. Jika dilihat dari kesemua
pengkhususan, purata pendapatan kelihatan seimbang antara satu sama lain
dan pendapatan ini adalah dalam bentuk tunai.
178
Jadual 2. Pendapatan dan purata pendapatan bulanan hasil daripada
perkhidmatan mengikut pengkhususan (n=289)
Pengkhususan
Jumlah pendapatan
Jumlah
Purata
bulanan (RM)
pengamal
pendapatan
(RM)
Umum
112,183.40
141
796
Perbidanan
34,340
49
701
Khusus
39,850
51
781
Campuran
35,510
48
740
Jumlah
221,883.40
289
768
KESIMPULAN
Kajian mendapati amalan perubatan tradisional Melayu (PTM) masih lagi
diamalkan dan dipelopori oleh golongan wanita. Kebanyakan masih lagi bergiat
aktif di kawasan luar bandar. Perkhidmatan di dalam bidang pengkhususan
perubatan umum masih menjadi tumpuan pengamal PTM berdasarkan
kepakaran mereka dalam merawat pelbagai penyakit ringan seperti demam,
kesihatan dalaman, batuk dan luka luaran. Pengamal PTM juga menghasilkan
produk sendiri berdasarkan pengetahuan turun-temurun dari segi menambah
nilai pendapatan isi rumah masing-masing. Pendapatan di antara keempatempat kelas pengkhususan perubatan tradisional Melayu menunjukkan tiada
perubahan yang ketara dari segi nilai tunai pendapatan sebulan. Khidmat
pengamal perubatan masih lagi diperlukan berdasarkan jumlah pesakit yang
diterima sebulan. Secara tidak langsung, ini menunjukkan masih ada lagi
pergantungan terhadap rawatan berasaskan perubatan tradisional Melayu
pada masa kini, walaupun kemajuan dari segi perubatan moden semakin
berkembang pesat. Sejajar dengan usaha meningkatkan nilai perubatan
tradisonal Melayu, usaha dokumentasi wajar diteruskan supaya maklumat
pengetahuan perubatan tradisonal masyarakat Melayu tidak hilang begitu
sahaja.
PENGHARGAAN
Ucapan penghargaan ditujukan kepada semua pengamal perubatan tradisional
Melayu(PTM) dan pegawai pengumpul data yang terlibat dalam menjalankan
survei dan juga pihak MOA yang membiayai dana penyelidikan.
179
PERALATAN PENYEDIAAN UBATAN DALAM KAEDAH RAWATAN
PERUBATAN TRADISIONAL MELAYU
AZ Wan Nurul Syafinaz, A Ida Farah, SM Ami Fazlin & BA Siti Khairul
Pusat Penyelidikan Perubatan Herba (HMRC), Institut Penyelidikan Perubatan,
Jalan Pahang, 50588 Kuala Lumpur, Malaysia
ABSTRAK
Salah satu elemen perawatan yang diamalkan oleh masyarakat Melayu ialah
rawatan herba yang mana kemujaraban ubat bergantung kepada bahan yang
digunakan serta cara penyediaan ramuan tersebut. . Peralatan yang sesuai di
dalam penyediaan bahan ramuan dari tumbuhan ubatan sangat penting bagi
mengoptimakan kemujaraban bahan ramuan tersebut. Kajian ini merekod
secara terperinci peralatan yang digunakan oleh pengamal perubatan
tradisional Melayu dalam penyediaan bahan ramuan untuk dijadikan ubatan
dalam merawat sesuatu penyakit. Pengamal dipilih melalui bancian yang
dijalankan melaui projek Pendokumentasian Secara Komprehensif
Pengetahuan Tradisional Melayu Berkaitan Tumbuhan Ubatan di Semenanjung
Malaysia berdasarkan jenis pengetahuan dan peralatan yang digunakan dalam
amalan mereka. Temu bual telah dijalankan dengan menggunakan borang kaji
selidik untuk mengumpul maklumat sejarah, pengalaman pengamal, kaedah
rawatan dan jenis peralatan yang digunakan dalam amalan rawatan mereka.
Peralatan diperolehi daripada pengamal semasa temu bual dijalankan atau
dibeli dari premis berkaitan. Seramai 65 orang pengamal ditemu bual bagi
kajian ini, dan 25 orang pengamal daripadanya menggunakan peralatan khusus
dalam penyediaan ubatan mereka. Sebanyak 21 jenis peralatan penyediaan
tumbuhan ubatan telah dikumpul sepanjang kajian ini. Penyediaan bahan
ramuan ubatan untuk merawat membabitkan proses termasuk menumbuk,
memipis, menggiling, mengasah, merendam, mencampur, meramas, dan
merendang. Peralatan yang dilaporkan dalam kajian ini terbahagi mengikut
kegunaannya untuk memproses bahan ramuan atau dalam proses
penyampaian rawatan itu sendiri. Pendokumentasian ini dapat merekod
amalan tradisional dalam penyediaan tumbuhan ubatan untuk digunakan
dalam perubatan tradisional Melayu. Kajian ini juga dapat memelihara ilmu
perubatan tradisional Melayu daripada lenyap ditelan zaman.
Kata kunci: peralatan, tumbuhan ubatan, perubatan tradisional Melayu
180
PENGENALAN
Tumbuhan ubatan berfungsi dalam menghindari atau merawat sesuatu
penyakit. Bahagian-bahagian tertentu pada tumbuhan atau keseluruhan
tumbuhan ubatan itu boleh dijadikan bahan ubatan; sebagai ekstrak bahan
aktif, bahan antibiotik, makanan dan rempah bagi tujuan perubatan atau tonik
untuk kesihatan (Sofowora 1982).
Peralatan yang digunakan dalam
penyediaan tumbuhan ubatan dan penyampaian rawatan itu sendiri amat
penting kerana peralatan tersebut boleh mempengaruhi kemujaraban sesuatu
ramuan. Di dalam kitab Tib iaitu manuskrip perubatan, peralatan yang
digunakan selalu diberi penekanan oleh penulis. Peralatan dalam penyediaan
herba adalah digunakan untuk menyukat, memproses, memasak dan
menyimpan bahan-bahan yang digunakan (Razak 2006). Bahan-bahan mentah
diproses sehingga terhasilnya bahan akhir seperti jamu, pilis, tapal, param,
minyak, ubat serbuk, serta ubat serbuk yang dijadikan pil atau kapsul.
Beberapa cara digunakan oleh masyarakat Melayu bagi menyediakan ramuan
ubat daripada tumbuhan ubatan. Antara proses yang terlibat dalam
penyediaan bahan ramuan ubatan adalah menumbuk, memipis, menggiling,
mengasah, merendam, mencampur, meramas, dan merendang.
BAHAN DAN KAEDAH KAJIAN
Satu bancian telah dijalankan ke atas pengamal perubatan tradisional Melayu
di Semenanjung Malaysia di bawah projek Pendokumentasian Secara
Komprehensif Pengetahuan Tradisional Melayu Berkaitan Tumbuhan Ubatan di
Semenanjung Malaysia. Pengamal ditemu bual berkaitan jenis amalan, tahap
pengetahuan serta peralatan yang digunakan. Seramai 65 orang pengamal
telah dipilih daripada senarai pengamal yang dibanci, berdasarkan amalan dan
peralatan yang digunakan. Borang kaji selidik digunakan untuk menemuramah
pengamal secara berstruktur dan terperinci mengenai amalan, pengetahuan,
sejarah dan peralatan yang digunakan. Peralatan digunakan oleh pengamal
yang direkod diperolehi sama ada daripada pengamal itu sendiri atau dibeli
dari premis yang berkaitan.
Daripada 65 orang yang telah ditemu bual bagi kajian ini, seramai 25 orang
pengamal menggunakan alatan khusus dalam penyediaan ubatan. Pengamal
menggunakan peralatan sama ada dalam proses penyediaan bahan ramuan
atau ketika rawatan dijalankan. Pengamal perubatan Melayu menggunakan
beberapa cara dalam proses penyediaan bahan ramuan daripada tumbuhan
181
ubatan. Sebagai contoh; rebus, celur, uli, asah, goreng, tumbuk, jemur, giling,
tanak, rendang, salai, bakar, keringkan, ramas dan rendam (Fatan 2003).
Sebanyak 21 jenis peralatan telah dikumpul sepanjang kajian ini
dijalankan (Jadual 1). Peralatan yang dikumpul dibahagikan kepada empat cara
penggunaan iaitu peralatan menyukat (Jadual 2), peralatan memproses (Jadual
3), peralatan memasak (Jadual 4) dan peralatan menyimpan (Jadual 5). Antara
contoh penggunaan peralatan dalam proses penyediaan bahan ramuan adalah
semasa menghasilkan pilis atau tapal. Lesung batu digunakan untuk
menumbuk rempah ratus sehingga hancur untuk dijadikan produk akhir. Satu
daripada contoh peralatan penting dalam perubatan tradisional ialah periuk.
Peralatan ini digunakan untuk merebus akar kayu, pucuk kayu, kulit kayu, daun
kayu serta bahan ramuan lain. Periuk juga digunakan sebagai bekas untuk
menyimpan ramuan yang telah siap diproses untuk kegunaan semasa rawatan.
Namun begitu, kebanyakan pengamal pada masa kini lebih cenderung
menggunakan teknologi moden seperti alat pengisar untuk menggantikan
peralatan tradisional kerana penggunaan mesin moden lebih menjimatkan
masa dengan hasil yang sama.
Jadual 1. Peralatan yang dikumpulkan dalam kajian
Proses
Jumlah Alatan
Menyukat
2
Memproses
7
Memasak
8
Menyimpan
4
JUMLAH
21
Jadual 2. Peralatan menyukat
Peralatan
Kegunaan
Cupak
Menyukat bahan ramuan yang digunakan untuk
membuat ubatan.
Gantang
Menyukat bahan ramuan yang digunakan untuk
membuat ubatan.
182
Jadual 3. Peralatan memproses
Peralatan
Kegunaan
Lesung batu
Menumbuk bahan ramuan supaya hancur atau
lumat dalam kuantiti yang sedikit.
Lesung kayu
Menumbuk bahan ramuan supaya hancur atau
lumat untuk kuantiti yang besar.
Piring asah
Mengasah bahagian tumbuhan ubatan untuk
ubat / piring
mendapatkan hasil serbuk atau air daripada asahan
kayu asah ubat tersebut.
Pasu asah ubat Mengasah bahagian tumbuhan ubatan untuk
mendapatkan hasil serbuk atau air daripada asahan
tersebut
Batu giling
Menggiling/memipis bahan ramuan sehingga
hancur/lumat.
Kacip
Memotong/mengacip bahagian tumbuhan ubatan
mengikut ukuran yang ditetapkan.
Jadual 4. Peralatan memasak
Peralatan
Kegunaan
Periuk / Periuk
Memasak/merebus bahan-bahan ramuan yang
Tinggi
telah diproses.
Periuk tanah /
Mangkuk tanah
telah diproses.
Periuk tembaga
telah diproses.
Periuk ubat
Merebus bahagian tumbuhan ubatan, dijadikan
air rebusan.
Kuali
Merendang/menggoreng bahan ramuan yang
telah diproses
Kuali tembaga
Merendang/menggoreng bahan ramuan yang
telah diproses
Kawah
Menanak minyak dengan skala yang besar.
Senduk/Sudip
Mengacau bahan ramuan semasa memasak.
Jadual 5. Peralatan menyimpan
Peralatan
Kegunaan
Botol kaca
Menyimpan ubatan yang sudah siap diproses.
Labu sayong
Menyimpan air penawar.
Mangkuk tanah
Menyimpan air rebusan.
Botol pil
Menyimpan pil kapsul yang sudah diproses.
183
KESIMPULAN
Peralatan yang digunakan dalam perubatan Melayu adalah sangat penting,
untuk memastikan kemujaraban ubat yang dihasilkan. Kajian mendapati para
pengamal masih lagi mengamalkan penggunaan peralatan tradisional Melayu
untuk menghasilkan ubat. Sungguhpun demikian, juga didapati pengamal pada
masa kini lebih cenderung menggunakan teknologi moden seperti alat pengisar
untuk menggantikan peralatan tradisional kerana penggunaan mesin moden
lebih menjimatkan masa dengan hasil yang sama.
PENGHARGAAN
Setinggi penghargaan diucapkan kepada kumpulan bancian projek ini,
pengamal yang memberi kerjasama untuk ditemu bual dan institusi- institusi
lain yang menyumbang dalam kajian ini. Projek dibiayai oleh Kementerian
Pertanian dan Industri Asas Tani.
RUJUKAN
Abdul Razak, A.K. (2006). Analisis Bahasa Dalam Kitab Tib Pontianak.
Perpustakaan Negara Malaysia, Kuala Lumpur. 192pp.
Fatan. H.Y. (2003). Potensi Tumbuh-Tumbuhan Sebagai Ubat-Ubatan. Jurnal
Fakulti Sains Sosial dan Kemanusiaan: 1─13.
Sofowora, A. (1982). Medicinal Plants and Traditional Medicine in Africa. John
Wiley & Sons, Chochester.
184
PHYSICO─CHEMICAL PROPERTIES OF SPRAY DRIED CLINACANTHUS
NUTANS EXTRACT
A Mohd. Suhaimi & AL Abd. Rahman
Food Science & Technology Research Centre, MARDI Headquarters, Persiaran
MARDI-UPM, 43400 Serdang, Selangor
Tel: 03-8953 6409
Fax: 03-8953 6500
ABSTRACT
Clinacanthus nutans or Sabah snake grass (SSG) powder was produced using an
Anhydro Lab S1 spray dryer. The powders were produced using three different
extract formulations as practised by You Dun Chao Healthcare Products.
Maltodextrin was blended with the extract and acts as an encapsulating agent.
The spray drying parameters were air inlet temperature of 1500C and outlet
temperature ranging from 75 to 850C, feed pump speed of 250 rpm and
compressed air pressure for atomization of 0.05 MPa. The SSG powders
produced were tested for water activity, bulk density, colour (L*, a*, b*),
moisture content, protein, oil content, ash and crude fibre. Results showed
that the ash, protein, oil and fibre contents were about the same as the
amount found in most salad herbs of Malaysia. Colorimetric analyses showed
that the L*, a* and b* values of the SSG powder showed significant differences
(p<0.05) among the formulations.
Keywords: Clinacanthus nutans, maltodextrin, encapsulating agent, spray
drying parameters
INTRODUCTION
Sabah snake grass or “belalai gajah”, also known scientifically as Clinacanthus
nutans is an important medicinal plant in Thailand (National Drug Comittee,
2006) and is widely used as a traditional medicine (Tiangburanatam 1996).
According to the World Bank report, trade in medicinal plants, botanical drug
products and raw materials is growing at an annual rate between 5 and 15%
(World Health Organization 2000). This indicates a good prospect for the Sabah
snake grass, as a medicinal plant to penetrate the market. A few claims on the
beneficial effects of Clinacanthus nutans leaves extracts are known, such as an
antidote for snake bites and its anti-inflammatory property. Currently, the
leaves are consumed fresh, processed into liquid extracts or dried in sachets.
Conversion of Sabah snake grass leaves extract into powder using a spray
drying method would make it convenient to be consumed. Dried extracts have
185
several advantages over the liquid form for industrial purposes, such as high
stability, easy to be standardized, transported and stored (Oliveira et al. 2006).
It also facilitates the manufacturing of solid dosage forms in the production of
tablets or capsules, which represent most of the medicines used worldwide
(Leuenberger & Lanz 2005).
Spray drying is a dehydration process of aqueous product. It converts
the liquid product into dry powder (Reineccius 1988). In this process, the
aqueous product will pass through an atomizer to generate fine droplets. Two
types of atomizer that are commonly used are the rotary disc atomizer and the
pressure nozzle. The droplets are contacted with circulated hot air in a drying
chamber and water starts to evaporate from the droplet surface. The droplets
are rapidly dehydrated, producing dried particles that are continuously
discharged from the drying chamber. There are three modes of hot air drying
movement: co-current flow, counter-current flow and mixed flow. The
advantages of using a spray drying method include high productivity and
increased microbiological stability of the product, especially for food. Spray
drying has been used to produce biomaterials and biopharmaceutical products
(Teixeira et al. 2011).
The spray drying process requires a drying aid to facilitate drying. The most
common drying aid used is maltodextrin (Bhandari et al. 1997). It helps to
reduce the agglomeration problems during storage, therefore improving
product stability (Bhandari et al. 1993; Silva et al. 2006). Maltodextrin is a
starch hydrolysate prepared by partial hydrolysis of corn starch or potato
starch using suitable acids and enzymes (USFDA). It is also classified as material
that has a dextrose equivalent of 3 to 20 with characteristics of no sweetness,
bland and not masking other flavours. Dextrose equivalent refers to the
percentage of reducing sugar in syrup calculated as dextrose on a dry weight
basis.
The aim of the study was to produce Sabah snake grass powder using a
spray dryer and evaluating its physico-chemical properties.
Raw Materials
The Sabah snake grass (SSG) leaves were obtained from You Dun Chao Herbs
Farm Sdn. Bhd while the extract was prepared by You Dun Chao Healthcare
Products Sdn. Bhd for spray drying. The maltodextrin was donated by ITS
Interscience Sdn. Bhd, Shah Alam, Selangor.
186
Spray Drying Procedures of SSG
The SSG extract was blended with 10-12 DE maltodextrin. The total soluble
solid of the mixture was measured using a hand held refractometer (Atago,
Master-alpha).The feed mixtures were stirred constantly to ensure feed
homogeneity during spray drying. The mixture was spray dried using Lab 1 APV
spray drier with inlet temperature at 150oC resulting with an outlet
temperature of 75oC to 85oC. The feed was pumped through the pressure
nozzle atomizer with a counter-current hot air movement produced by the
heater and the blower. The compressed air pressure for atomizing was set at
0.05 MPa.
SSG powder analysis
1. Moisture content
The moisture content was determined by drying the SSG powder in an oven at
1050C overnight. Drying was continued until a constant reading was obtained.
The weight of the SSG powder before and after the drying process was
recorded. The moisture content of the powder was calculated.
2. Water activity
Water activity of SSG powder was measured using a water activity meter
(Aqualab Series 3, USA).
3. Bulk density
Bulk density of the powder was measured by weighing 10 g of sample and
placing it in a 100 ml graduated cylinder. A steady vibration was conducted on
a vibrator (Main et al. 1978). The volume was then recorded and used to
calculate bulk density as g/ml.
4. Colour (L, a, b)
Colour was measured using a colour reader (Minolta chromameter, model CR
300/colorimeter series). The values of L (Lightness), a (redness) and b
(yellowness) were determined.
5. Protein.
Crude protein was determined according to the Kjeldahl method using the
conversion factor of 6.25 (AOAC, 1990).
6. Oil content
Oil content was determined according to AOAC method 920.39C (AOAC, 1990).
187
7. Crude fiber
Crude fiber was determined using the fibretec system (according to Weende).
8. Ash
Ash content was determined according to AOAC method 923.03 (AOAC 1990).
Statistical analysis
Three formulations, SSG3 (addition of lemon juice), SSG5 (normal) and SSG8
(concentrated) were analysed in triplicate. Analysis of variance was carried out
to differentiate the means of the variables for the samples. The results were
expressed as mean values  standard deviation (SD).
Physicochemical properties
Moisture content of SSG powder ranges from 0.51 to 0.91% (Table1) and no
differences (p>0.05) in moisture content were detected between formulations.
The least amount of moisture shows the rate of heat transferred to the
particles was high. The production of low moisture content powder is
important in spray drying where food powder with 10% moisture content helps
to retard mould growth (Hagan 2007).
Water activity for all powders were less than 0.5. According to Quek et
al. (2007), food that has water activity less than 0.6 is classified as
microbiologically stable. This indicates the powder might have longer shelf life.
The bulk density of the powder varied from 0.37 to 0.42 g/ml. A high
drying temperature would have caused higher surface ratio to volume for the
spray dried powder particles. In addition, the lesser amount of compressed air
for atomizing may produce larger particle size of the powder. This may lead to
lower bulk density of the powder. The larger the size of the particles would
have resulted in more trapped air between them. Thus, leading to a greater
possibility for oxygen degeneration of the pigment and reducing storage
stability.
The percentage of protein, fiber, ash and oil content in all the samples
were about the same as the amount found in most salad herbs of Malaysia as
reported by Wan Hassan and Mustaffa (2010).
188
Table 1. Physicochemical composition of spray dried SSG powder
Variables
SSG 3
SSG 5
SSG 8
(Addition
of (Normal)
(Concentrated)
lemon juice)
Moisture content (%) 0.51 ± 0.32a
0.59 ± 0.06a
0.91 ± 0.52a
Water activity, aw
0.21 ± 0.01c
0.30 ± 0.01b
0.35 ± 0.02a
b
a
Ash (%)
0.57 ± 0.01
0.84 ± 0.11
0.76 ± 0.05a
b
a
Protein (%)
0.08 ± 0.00
0.16 ± 0.02
0.16 ± 0.01a
Oil content (%)
5.35 ± 2.74ab
10.75 ± 5.27a 2.11 ± 1.10b
a
Fibre (%)
0.51 ± 0.24
0.56 ± 0.29a
0.45 ± 0.12a
Bulk density (g/ml)
0.37 ± 0.14a
0.39 ± 0.05a
0.42 ± 0.01a
*Means with the same letter are not significantly different (for variables
Colour
The colour on the SSG powder was measured (Figure 1). The L*, a*, and b*
values showed significant differences (p<0.05) among the formulations. The
highest value for L* was given by SSG 3 indicating lighter product towards
whiteness. This might be due to the addition of lemon juice that impaired the
green pigment in the extract. SSG 5 had a* negative value corresponding to a
greenness although the extract is more diluted compared to SSG 8. SSG 5 had
the highest b* value. This indicates that the yellowness of the product was not
influenced by the lemon juice in SSG 3.
Figure 1: Colour measurement of spray dried SSG powder
189
CONCLUSION
Sabah snake grass powder was obtained using 10% maltodextrin at inlet
temperature of 150oC. Addition of lemon juice in the extract helps to mask the
unpalatable taste in normal extract of SSG. Further study is required to look
into its potential as a health supplement.
ACKNOWLEDGEMENTS
The author thanks Mr. Hazmi bin Haron and Mr. L.E. Chong from You Dun Chao
Healthcare Products & You Dun Chao Herbs Farm Sdn. Bhd for the supply of
Sabah snake grass extract.
REFERENCES
AOAC. (1990). Official Methods of Analysis of the Association of Official
Analytical Chemists, 15thed. Arlington, VA: Association of Official
Analytical Chemists.
Bhandari, B.R., Snoussi, A., Dumoulin, E.D. & Lebert, A. (1993). Spray Drying of
Concentrated Fruit Juices. Drying Technology 11(5):1081–1092.
Bhandari, B.R., Datta, N. & Howes, T. (1997). Problems Associated with Spray
Drying of Sugar-Rich Foods. Drying Technology 15 (2): 671–684.
Hagan, A.T. (2007). Part 1X of Food Storage FAQ. Shelf Reliance. http://
www.shelfreliance.com/library/view/57 (accessed April, 2009).
Leuenberger, H. & Lanz, M. (2005). Pharmaceutical Powder Technology – From
Art to Science. The Challenge of the FDA´s Process Analytical
Technology Initiative. Advanced Powder Technology 16(1): 3–25.
Main, J.H., Clydesdale, F.M. & Francis, F.J. (1978). Spray Drying Anthocyanin
Concentrates for Use as Food Colorant. J. Food Sci. 43: 1693–1694.
National Drug Committee. (2006). List of Herbal Medicinal Products A.D. 2006.
ISBN: 974-244-217-7, Chuoomnoom Sahakorn Karnkaset Publisher,
Bangkok, Thailand. Pp. 59–61.
Oliveira, W.P., Bott, R.B. & Souza, C.R.F. (2006). Manufacture of Standardized
Dried Extracts from Medicinal Brazilian Plants. Drying Technology
24(4): 523–533.
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Quek, S.Y., Chok, N.K. & Swedlund, P. (2007). The Physicochemical Properties
of Spray Dried Watermelon Powder. Chemical Engineering and
Processing 46(5): 386–392.
Reineccius, G.A. & Risch, S.J. (1988). Spray Drying of Food Flavors. In Flavor
Encapsulation. Washington D.C: American Chemical Society. Pp. 55–66.
Silva, M.A., Sobral, P.J.A. & Kieckbusch, T.G. (2006). State Diagrams of FreezeDried Camu-Camu (Myrciaria dubia (HBK) Mc Vaugh) Pulp With and
Without Maltodextrin Addition. Journal of Food Engineering 77(3):
426–432.
Teixeira, C.C.C., Teixeira, G.A. & Freitas, L.A.P. (2011). Spray Drying of Extracts
from Red Yeast fFrmentation Broth. Drying Technology 29: 342–350.
Tiangburanatam, W. (1996). Dictionary of Thai Medicinal Plants, 4th ed.
Prachoom Karn Pim, Bangkok, Thailand. Pp. 788–789.
USFDA. (2006). Alphabetical List of SCOGS Substances. http://www.
cfsan.fda.gov/~dms/opascogc.html.
Wan Hassan,W.E. & Mustaffa, M. (2010). Ulam Species. In Ulam: Salad Herbs of
Malaysia. Masbe Publisher, Kuala Lumpur, Malaysia. Pp. 40–239.
World Health Organization. (2000). Report of the Inter-Regional Workshop on
Intellectual Property Rights in the Context of Traditional Medicine,
Bangkok, 6–8 December 2000.
191
EXTRACTION OF ANTIOXIDANTS FROM LEAVES OF CLINACANTHUS
NUTANS (BURM.F.) LINDAU: EFFECTS OF EXTRACTION METHOD AND
SOLVENT
NA Karim1 & II Muhamad 1,2
1
Department of Bioprocess Engineering, Faculty of Chemical Engineering,
Universiti Teknologi Malaysia (UTM), Johor Bahru, 81310 Johor
2
IJN-UTM Cardiovascular Engineering Centre, Faculty of Bioscience and
Medical Engineering, Universiti Teknologi Malaysia, Johor Bahru, 81310 Johor
Tel: 07-5535577/ 07-5558564 Fax: 07-5588166 E-mail: [email protected]
ABSTRACT
The aim of this study was to determine the best method for extracting
antioxidants from Clinacanthus nutans (Burm.f.) Lindau, (C. nutans) leaves. The
extraction was done by four different methods: decoction (100ᵒC, 20 min),
soxhlet (8 hour), maceration (3 day) and ultrasound-assisted extraction (25ᵒC,
40 min) using different solvents: ethanol, methanol and water respectively.
Antioxidant activity, total phenol, and flavonoid content were determined from
the various extraction methods. Total phenolic and ﬂavonoid contents were
determined by Folin–Ciocalteau and aluminum chloride methods,
respectively. Antioxidant activity was detected by 2,2-diphenyl-1-picrylhydrazyl
(DPPH) free radical scavenging activity and reducing capacity assessment
(FRAP). The results showed that the extraction method and solvent used
signiﬁcantly affect the extraction of antioxidant from the leaves. Conventional
extraction method (water extracts) proved to be the best method with
maximum yield (31.60 ± 0.44 %, w/w), total phenolic (24.76 ± 3.39 mg gallic
acid equivalent per g dry weight sample) and flavonoid content (6.25 ± 0.42 mg
quercetin equivalent per g dry weight sample). It also showed strongest DPPH
free radical scavenging activity (16.29 ± 2.69 mg trolox equivalent antioxidant
capacities (TEAC)/g dry weight sample) and FRAP assay (19.40 ± 0.24 TEAC/g
dry weight sample). As a conclusion, C. nutans leaves are potential natural
antioxidant for the development of nutraceutical and functional food.
Keywords: Clinacanthus nutans, natural antioxidant, flavonoid, decoction
INTRODUCTION
In recent years, the usage of herbs and their extracts complimented
modern medicine in hospitals as well as being introduced as dietary
supplements. A new concept in the usage of naturally derived chemicals from
nature which contained high antioxidant and phytochemical properties has
drastically advanced the field of natural products. It showed their application
192
potential in the treatment and prevention of cancer, cardiovascular and other
chronic diseases (Karabegovic et al. 2014). Clinacanthus nutans (Burm.f.)
Lindau (C. nutans), belonging to the family of Acanthaceae is native to the
Southeast Asia region and is cultivated in Thailand, Malaysia and Indonesia. It is
an edible erect herbal plant, characteristically described as a finely pubescent
scrub with a drooping and clambering trait (Quattrocchi 2012). It has been
reported to have biological activities and high medicinal values, including
antioxidant activity (Yong et al. 2013).
The extraction techniques play an important role in the extraction of
beneficial bioactive compounds or drugs from natural plant matrices (Azmir et
al. 2013; Sulaiman et al. 2015). Conventional extraction techniques such as
decoction, soxhlet and maceration are still widely practiced even though
ultrasound-assisted extraction is one of the new extraction methods capable of
providing efficient cell disruption for high quality extract and is also energysaving (Wang & Weller 2006). Different solvent system could give different
extraction yield, antioxidant activity and chemical compositions. Theoretically,
the optimal extraction technique and choice of solvent should be simple, safe,
reproducible, inexpensive, allow complete extraction of the interest
compounds, not destructive and suitable for industrial application (Karabegovic
et al. 2014). Therefore, water, ethanol and methanol are commonly used
solvent in extraction. The aim of this study was to investigate the effects of
different extraction methods (soxhlet, maceration, ultrasound-assisted
extraction and decoction) employed in three extracting solvents (ethanol,
methanol and water), on the extraction yields, antioxidant activity, total
phenolic, and flavonoid content of C. nutans leaves extracts.
Chemicals and Reagents
Chemicals and reagents used were Folin–Ciocalteau’s phenol reagent and
anhydrous sodium sulphate (Merck), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and
acetic anhydride (Sigma Aldrich), Trolox, 97% (ACROS Organics), sodium
carbonate anhydrous (Fluka Chemical Corp) and absolute ethanol (99% purity,
RCI Labscan Limited). All other chemicals were analytical grade.
Collection and Preparation of Plant Materials
The leaves and stem of C. nutans were collected in December, 2014 from TKC
Herbal Nursery, Seremban, Negeri Sembilan, Malaysia. The plant material was
botanically identified by Dr. Shamsul Khamis (Biodiversity Unit, Institute of
Bioscience, Universiti Putra Malaysia, Malaysia) with specimens voucher
193
number SK2781/2015. The leaves and stems were separated and washed
thoroughly with tap water, oven dried at 50oC homogenized to fine powder.
They were kept in an airtight plastic 4°C until further extraction.
Extraction of C. nutans Leaves
The powdered C. nutans leaves or stem were extracted using different
extraction methods (soxhlet, maceration, ultrasonic and decoction methods)
involving the use of three types of solvents (ethanol, methanol and distilled
water) with the ratio of plant material-to-solvent at 1:10 (w/v). All experiments
were performed in triplicate. For soxhlet extraction (SE), powdered C. nutans
leaves (15 g) were extracted individually with ethanol, methanol and water
(150 ml) for 8 hours in a soxhlet apparatus (250 ml boiler) with controller
heating mantles, respectively. For maceration extraction (ME), leaves were
soaked with respective solvent in a closed beaker and were left at room
temperature for 3 days. Ultrasonic-assisted extraction (UAE) was performed for
40 min, at 25○C with a 20 min time interval respectively in an ultrasonic cleaner
(LIR™ Biotech, JP-040S, Malaysia). Decoction extraction (DE) was done by
heating and boiling the leaves for 10 min. The decoction was rapidly cooled
under tap water before being centrifuged at 10,000 rpm for 10 min. After the
extraction process, the extract was collected and filtered using filter paper. The
collected sample was concentrated by rotary evaporator (Heidolph, Laborota
4000, Switzerland) at temperature 40○C to obtain the crude extract (extraction
yield). The extraction yield was determined using the following equation:
Extraction Yield (%, w/w) = Me (g)/ Ms (g) x 100, where Me is mass of extracts,
and Ms is mass of dry sample. Then, the residue obtained was kept at -20 ○C
until further analysis.
Total Phenolic and Flavonoid Content Determination
Folin–Ciocalteau method was used to determine total phenolic content (TPC) in
sample extract (Waterhouse 2002). Total flavonoid content (TFC) was
determined using colorimetry of aluminium chloride method as described by
Zhishen et al. (1999) with some modifications.
Antioxidant Assays
Total antioxidant activity was analysed using two assays: DPPH free radical
scavenging assay based on method by Abu Bakar et al. (2009) and ferricreducing antioxidant power assay (FRAP) according to the procedure of Benzie
and Strain (1996). Trolox (0.02 – 0.10 mg/ml) was used as reference standard.
The antioxidant activity was expressed as mg of Trolox equivalents antioxidant
capacity (TEAC) per gram of plant material on a dry weight as it showed
accurate and descriptive expression than assays that express antioxidant
194
activity as the percentage decrease in absorbance. The results provide direct
comparison of the antioxidant activity with Trolox.
Extractive Yield
The extractive yields from C. nutans leaves extracts obtained by soxhlet,
maceration, ultrasonic and decoction using ethanol, methanol and water,
respectively were presented in Figure 1. Results showed the extraction
methods, solvents and plant parts strongly affected the extractive yield. The
boiling water (decoction) showed the best extraction efficiency with the
highest extractive yield of 31.6 ± 0.44 %, w/w, followed by soxhlet, maceration
and ultrasonic method. In Figure 1, water (aqueous) extracts obtained by all
discussed methods showed greater extractive yield than methanol and
ethanol. Generally, the yield obtained corresponded to the sequence of
polarity: water > methanol > ethanol. Similar findings were reported by Cabana
et al. (2013) and Bampouli et al. (2014), where the aqueous extracts exhibited
the highest extractive yield as compared to alcoholic extracts. In the case of
alcoholic extracts, methanol extracts showed the highest yield than ethanol
extracts regardless of extraction methods. The differences between yields of
the studied extracts were due to the polarity of the compounds present in the
plant materials.
Figure1. Extractive yield of C. nutans leaves obtained by different extraction
methods and solvents
Total Phenolic and Flavonoid Contents
The total phenolic content (TPC) and total flavonoid content (TFC) determined
in C. nutans leaves extracts obtained by different extraction methods and
solvents were depicted in Figure 2. The TPC varied from 3.88 to 44.76 mg of
GAE/g of dry mass sample and the TFC ranged from 1.33 to 12.22 mg of QE/g
of dry mass sample. The aqueous extracts of C. nutans obtained by all studied
methods had highest TPC values (ranging from 30.24 to 44.76), as compared to
195
alcoholic extracts (TPC: water > methanol > ethanol). From each different
extraction methods tested, soxhlet and decoction methods showed higher TPC
value with 6.51 to 45.19 and 9.48 to 44.76 mg GAE/g dry weight sample,
respectively. These results showed that phenolic contents were strongly
dependent on polarity of solvents used, which polar fractions (water) had more
phenolics in them than the less-polar fractions (methanol/ethanol) (Hayouni et
al. 2007). The TFC results showed a contradict trend as compared to TPC. The
ethanolic extracts exhibited a higher amount of TFC as compared to methanolic
and aqueous extracts in soxhlet method (TFC: ethanol > methanol > water).
These could be attributed to the solubility of flavonoid compounds which is
soluble in alcoholic solvents. The flavonoid content in C. nutans leaves could be
from flavones C-glycosides (shaftoside, isomollupentin 7-O-𝛽-glucopyranoside,
orientin, isoorientin, vitexin, and isovitexin) as previously reported by Teshima
et al. (1998). It was also reported that compounds such as flavonoid, which
contain hydroxyl functional groups, are responsible for antioxidant effects in
the plants (Jaberian et al. 2013).
Figure 2. Total phenolic (A) and total flavonoid contents (B) of C. nutans leaves
by different extraction methods and solvents
Total Antioxidant Activity
The results on total antioxidant activity (TAA) measured by DPPH and FRAP
assay using standard Trolox were summarized in Figure 3. As shown in Figure 3,
TAA values varied from 0.74 to 16.29 and 1.39 to 29.58 mg of TEAC/g of dry
mass sample with DPPH and FRAP assay, respectively. TAA values of C. nutans
showed similar trend as TPC and TFC value in which leaves extract has higher
TAA than stem extract for each extraction methods and solvents. The higher
antioxidant activity exhibited by the aqueous extracts (decoction and soxhlet)
over other solvents and methods clearly demonstrates the relative advantage
of green extraction method using water as solvent that managed to extract
maximum antioxidants. From the results of the DPPH and FRAP assays, the
decoction and soxhlet extracts (aqueous) of C. nutans showed significant
inhibitory activity against free radicals, while it showed comparatively less
196
activity in sonication and maceration. Heat treatment applied in soxhlet and
decoction methods increased the concentration of antioxidant in C. nutans, in
which the heat treatment enhanced diffusivity, soften the plant tissues and
promoted elution of attached bioactive compounds (Cabana et al. 2013).
Figure 3. Total antioxidant contents of C. nutans leaves by different extraction
methods and solvents obtained by two different assays: (A) DPPH and (B) FRAP
CONCLUSION
Decoction method (aqueous extracts) of C. nutans was found to be the best
extraction method and solvent as it showed high extraction efficiency and
strong antioxidants in term of high TPC and TFC values. In addition, C. nutans
leaves also showed significantly higher antioxidant contents for both DPPH and
FRAP assays. Therefore, the traditional decoction method was still a reliable
extraction process for and was considered as environmental-friendly, simple,
safe, reproducible and inexpensive method for extracting antioxidant.
ACKNOWLEDGEMENTS
We are grateful to the Ministry of Agriculture and Agro-based Industry,
Malaysia for the NKEA Research Grant Scheme (NRGS) (vote 4H023) and the
Ministry of Higher Education (MOHE) for the MyBrain15 scholarship for PhD
scholarship awarded to Norsuhada Abdul Karim.
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199
ANTI-PROLIFERATIVE EFFECT OF CLINACANTHUS NUTANS
OVARIAN, BREAST AND COLORECTAL CANCER CELL LINES
ON
Y Nurhanan Murni, SK Ling, SM Siti Syarifah, A Zunoliza, MY Nor Afiedatul
Akmal
Forest Research Institute Malaysia, 52109, Kepong, Selangor
Tel: 03-6279 7659
Fax: 03-6280 4623
ABSTRACT
Clinacanthus nutans (Acanthaceae) is locally known as belalai gajah or Sabah
snake grass. It has been traditionally used in several Asian countries as antiinflammatory agent and to treat dysentery and viral infection. The species is
also popularly sought for its use as anti-cancer remedies. In this study, an in
vitro anti-cancer model utilising ovarian (SKOV-3), breast (MCF-7) and
colorectal (HT-29) cancer cell lines and Sulphorhodamine B (SRB) assay were
used to investigate the inhibitory effects of the leaves of C. nutans on the
proliferations of these cancer cell lines. The results showed that the
methanolic, hexane, ethyl acetate and aqueous extracts of the leaves gave
lower anti-proliferative effects (IC50 range: 292.63 ± 11.58 g/mL to 359.17 ±
20.22 g/mL) than known anti-cancer drug cisplatin (IC50 range: 0.38 ± 0.01
g/mL to 0.77 ± 0.07 g/mL) when treated on SKOV-3 and MCF-7 cancer cell
lines. The hexane extract was further fractionated into five fractions (coded as
CLNH1,2,3,4,5) and it was found that one of its fractions, CLNH2 showed an
increase in anti-proliferative effect when treated on SKOV-3, MCF-7 and HT-29
cancer cell lines with IC50 values of 56.51 ± 3.42, 54.63 ± 2.33 and 41.54 ± 3.10
g/mL, respectively. Further study that includes isolation and identification of
compound(s) in C. nutans had been conducted. This paper also describes the
anti-proliferative activities of some of the major compounds isolated from C.
nutans on the respective cancer cell lines.
Keywords: Clinacanthus nutans, leaves, anti-proliferative, cancer cell lines,
Sulphorhodamine B assay
INTRODUCTION
Cancer still affected millions of people worldwide. It is estimated that one in
eight deaths is due to cancer and it causes more deaths than AIDS,
tuberculosis, and malaria combined globally (American Cancer Society 2011).
Breast and ovarian cancers still remain among top ten deadliest cancers among
women while colorectal cancer is among the deadliest cancer among men. In
Malaysia, cancer is the fourth leading diseases that cause death after
200
circulatory, respiratory and infectious diseases (Ministry of Health 2014).
Cancer starts from over proliferation of abnormal cells that can spread to other
parts of its origin and often diagnosed at late stage, thus may reduce chance of
survival. Often, the treatment used includes chemotherapy that is the use of
drug either alone or in combination with other drug or mode of treatment (i.e.
radiotherapy, immunotherapy, etc.). The role of chemotherapy is to kill the
cancer cells but sometimes this treatment is ineffective due to the toxic side
effects and drug resistance problems. Hence, the searches of new drug
candidate that can reduce these problems are in dire needs.
It is known that plants have been one of important resources for
bioactive or lead compounds in generating conventional medicines including
chemotherapy drugs (Newman & Cragg 2012) and Malaysia as one of the
twelve countries with megadiversity has approximately 15,500 plant species to
be explored on its medicinal properties. Clinacanthus nutans is known as
belalai gajah locally or bi phaya yaw in Thailand. The plant is native in most of
South East Asian countries to the southern China and the leaves had been
traditionally used to treat dysentery, ophthalmia (Burkill 1935) and also as
snake venom antidote (Levey 1969). The plants have also been reported to
have anti-viral effect against herpes infection (Kongkaew & Chaiyakunapruk
2011, Kunsorn et al. 2013), anti-microbial (Yang et al. 2013, Arullapan et al.
2014), anti-inflammatory (Wanikiat et al. 2008) and anti-oxidant (Yong et al.
2013) properties. C. nutans has gained popularity locally due to the plant is
claimed to treat various type of cancers but the scientific evidences on this
anti-cancer effects are still lacking. Hence, the aim of this study is to determine
the anti-cancer potential of the extracts and compounds isolated from C.
nutans by analysing its anti-proliferative effects on selected cancer cell lines.
General Procedure
1
H and 13C NMR spectra were recorded in ppm (δ) in CDCl3, CD3OD and DMSOd6 employing a Bruker DRX 300 spectrometer operating at 300 MHz for 1H and
75 MHz for 13C, respectively. Column chromatographies were performed with
silica gel 60 (0.040-0.063 mm, Merck, Germany), MCI gel CHP 20P (75-150 µm,
Supelco, USA), Diaion HP-20 (250-850 µm, Supelco, USA), Diaion HP-20SS (75150 µm, Supelco, USA), Chromatorex ODS (100-200 mesh, Fuji Silysia Chemical,
Ltd.) and Sephadex LH-20 (GE Healthcare Bio-Science AB, Sweden). TLC was
performed on precoated Si gel 60 F254 plates (0.2 mm thick, Merck) and spots
were detected by UV illumination and by spraying with 10% H2SO4 solution
followed by heating.
201
Plant Materials
The fresh aerial part of C. nutans was purchased from Pusat Pertanian Pantai,
Jalan Pantai Batu 7, Pantai, Seremban. The leaves were separated from the
stems. Then they were dried in an oven at 40–45°C, followed by grinding into
powder.
Preparation of Extract
The powder material was extracted four times by soaking in methanol (4 kg :
25 L) at room temperature and 3 days interval. After filtration, the extracted
portions were combined and concentrated by evaporation under reduced
pressure to give the crude extract. The methanol extract was suspended in
distilled water and partitioned successively with hexane and ethyl acetate to
give the hexane, ethyl acetate and water fractions. Further fractionation was
carried out on the hexane fraction using column chromatography with Diaion
HP-20SS as the matrix which resulted in 5 sub-fractions (F1-F5).
Fractionation and Purification
The hexane and water fractions were each subjected to a combination of
column chromatographies with various media such as MCI gel CHP 20P, Diaion
HP-20, Diaion HP-20SS, Chromatorex ODS, Sephadex LH-20 and silica gel, and
different solvent systems to yield seven thin layer chromatographically
homogeneous compounds; CNLW651 (1), CNLH44 (2), CNLH55A (3), CNLH55B
(4), CNSW1442 (5), CNSf4 (6) and CNSW (7). Compounds 5-7 were also purified
from the stems.
Cell Lines and Treatments
Ovarian (SKOV-3), breast (MCF-7) and colorectal (HT-29) cancer cell lines were
purchased from American Type Culture Collections, USA. The cell lines were
cultured and sub-cultured in Dulbecco’s Modified Eagle’s medium (DMEM)
supplemented with 5% fetal bovine serum (FBS), 1% penicillin-streptomycin,
0.25% amphotericin B and 1% gentamycin. Approximately 4000 to 6000 cells
were seeded in each well of the 96 well plates and incubated in a humidified
incubator at 37C and 5% carbon dioxide in air for 24 h. Each cell line was then
treated at five different concentrations of C. nutans’s extracts and fractions (1,
5, 25, 125, 625 µg/mL) and compounds (1, 10, 20, 50 100 µg/mL) in triplicate.
At least 24 wells were untreated with C. nutans in each plate. Cisplatin, a
known chemo-drug was also treated on these cell lines at different
concentrations (0.032, 0.16, 0.8, 4, 20 µg/mL) as for comparison studies. The
202
cells were then incubated in the same incubator with the mentioned
conditions for 72 h. The experiment was repeated at least three times.
Sulphorhodamine B (SRB) Assay
The 72 h treatments were stopped by performing Sulphorhodamine-B (SRB)
assay (Skehan et al. 1990). The assay was performed by firstly adding the 50 µL
of ice cold tricholoroacetic acid (TCA) for fixing the cells for 30 mins. Second
step involved with staining the cells with 100 µL of 0.4% SRB for 30 mins and
followed with rinsing the wells with 1% acetic acid. Finally, 100 µL of Tris buffer
was then added in each well and the results of the treated and untreated cells
were read at an optical density (OD) of 492 nm using a Magellan V.4 microtiter
plate reader (Tecan, Switzerland). The percentage of cells viability were then
calculated based on (OD492nm of the treated cells/ OD492nm of the untreated
cells) x 100. The IC50 values were determined from the dose-responsed curves
of percentage of cells viability versus the concentrations of the C. nutans
samples (µg/mL).
Identification of Compounds from C. nutans
The isolation and purification work on the hexane and water fractions of leaves
has resulted in the identification of 4 known compounds in the leaves as
shaftoside (1) (Osterdahl 1979, Kondo et al. 2000), stigmasterol (3), β-sitosterol
(4) (Vencata et al. 2012a) and lupenone (6) (Prachayasittikul et al. 2010).
Among these, lupenone was also obtained from the stems. Additionally,
structure analysis of three compounds, i.e., 2, 5 and 7, are still on-going.
CH3
H3C
CH3
CH3
H3C
CH3
H2C
CH3
CH3
CH3
CH3
CH3
CH3
CH3
CH3
CH3
CH3
HO
HO
HO
H3C
Shaftoside (1)
Stigmasterol (3)
203
-Sitosterol (4)
CH3
Lupeol (6)
Anti-proliferative Effects of Extracts, Fractions and Compounds from C.
nutans
The anti-proliferative effects of phytochemicals (extracts, fractions and
compounds) obtained from the leaves of C. nutans were analysed based on its
IC50 values (Table 1). The extracts and fractions of C. nutans showed antiproliferative effects at very high concentrations except one fraction CNLH2
showed moderate anti-proliferative effect when tested on HT-29 cancer cell
line. 2 out of 7 compounds (β-sitosterol and lupeol) also showed moderate
anti-proliferative activities when tested on these three cancer cell lines (SKOV3, MCF-7 and HT-29).
Stigmasterol and β-sitosterol that belongs to phytosterol group of
compounds are also common in many plant species particularly from the oil,
seeds, nuts and legumes. These phytosterols are used in diet and claimed to
protect cardiovascular diseases and have anti-cancer effect in colon cancer
(Awad & Fink 2000). Lupeol (also known as Fagarsterol) found in white
cabbage, green pepper, strawberry, olive, mangoes and grapes was also
reported to induce cell death in several cell lines including T-lymphoblastic
leukemia CEM (IC50 =50 μM), breast carcinoma MCF-7 (IC50 =50 μM], lung
carcinoma A-549 (IC50 =50 μM), multiple myeloma RPMI 8226 (IC50 =50 μM),
cervical carcinoma HeLa (IC50 =37 μM), and malignant melanoma G361 (IC50
=50 μM) (Saleem 2009). The presence of shaftoside in C. nutans detected by
HPLC analysis had also been reported by Chelyn et al. (2014) but the data on its
effect on cancer cells proliferation are still lacking. The compound was found to
be the least active in this anti-proliferative study (Table 1).
204
Table 1. The IC50 values of the C. nutans samples (µg/mL) treated in ovarian
(SKOV-3), breast (MCF-7) and colorectal (HT-29) cancer cell lines
Cancer Cell Line
SKOV-3
MCF-7
HT-29
C. nutans
Name/ Codes
samples
Extract
Methanol
292.63
± 359.17
± 330.40 ± 9.79
11.58
20.22
Hexane
311.38
± 320.11
± 341.39 ± 6.35
0.35
5.37
Ethyl Acetate
254.61
± 268.93
± 282.10 ± 11.65
33.69
16.11
Aqueous
284.58
± 297.03
± undetermined
18.02
4.07
Fractions
CNLH1
CNLH2
CNLH3
CNLH4
CNLH5
Compounds
Drug
CNLW651
(Shaftoside)
CNLH44
CNLH55A
(Stigmasterol)
CNLH55B
(β-sitosterol)
CNSW1442
CNSf4 (Lupeol)
311.12
11.45
56.51
3.42
382.22
19.85
356.20
8.38
337.62
16.38
± 339.18
12.73
± 54.63
2.33
± 402.28
3.96
± 349.75
1.44
± 358.14
13.23
>100
>100
CNSW
>100
77.55
0.68
35.52
1.46
>100
33.36
0.50
>100
>100
± 80.54
0.80
± 40.56
1.46
>100
± 34.09
1.61
>100
Cisplatin
0.57 ± 0.01
± 328.58 ± 7.31
± 41.54 ± 3.10
± 405.74 ± 5.81
± 379.01 ± 9.21
± 350.21 ± 8.04
>100
>100
± 82.84 ± 1.26
± 39.35 ± 0.99
>100
± 61.79 ± 0.57
0.56 ± 0.10
>100
0.48 ± 0.08
Inference: Sample that gave: IC50 ≤ 20 g/mL = very active; 20 g/mL IC50 ≤ 50 g/mL = moderately active;
50 g/mL  IC50 ≤ 100 g/mL = weakly active; IC50  100 g/mL = least active
205
CONCLUSION
Although C. nutans extracts showed anti-proliferative activities on respective
cancer cell lines, higher concentrations of C. nutans extracts that were
approximately 600 times than the drug cisplatin were needed to inhibit the
proliferation of at least 50% cells population in vitro. β-sitosterol and lupeol
showed highest anti-proliferative activities among the seven isolated
compounds. Further in vivo anti-cancer studies and toxicity evaluation on the
respective extracts and compounds shall be conducted to ensure its efficacy
and safety prior to pre-clinical and clinical tests towards development as anticancer agent.
ACKNOWLEDGEMENTS
The authors are grateful to Ministry of Agriculture and Agrobased Industry,
Malaysia for providing financial support and Ms. Ruzana Rabuzin for assisting in
performing the cell culture work.
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208
DETECTION OF IRRADIATED HERBS USING PHOTO-STIMULATED
LUMINESCENCE TECHNIQUE (PSL)
AR Ros Anita, O Zainon, A Foziah & H Zainab
Agotechnology & Bioscience Division, Malaysian Nuclear Agency, 43000 Kajang,
Selangor
Tel: 03-8911 2000 Fax: 03-8911 2157 E-mail: [email protected]
ABSTRACT
Photo-stimulated luminescence (PSL) technique was applied to detect
irradiated herbs such as mas cotek (Ficus deltoidea), misai kucing (Orthosiphon
stamineus) and hempedu bumi (Andrographis paniculata). Using screening and
calibrated PSL, all samples were correctly discriminated between nonirradiated and irradiated with dose 5 kGy. The PSL photon counts (PCs) of
irradiated herbs increased with increasing dose, with mas cotek showing
highest sensitivity index to irradiation compared to misai kucing and hempedu
bumi. The differences in response are possibly attributed to the varying
quantity and quality of silicate minerals present in each herbs sample. The
results of this study provide a useful database on the applicability of PSL
technique for the detection of Malaysian irradiated herbs.
Keywords: herbs, photo-stimulated luminescence (PSL), index of sensitivity,
calibrated PSL, irradiated food
INTRODUCTION
Food irradiation has been proven as one of the alternative techniques to the
existing methods for increasing the shelf-life, improving the hygienic quality,
and enhancing the functional properties of different food items (Akram et al.
2012). An important problem is to develop method for unambiguous
identification of irradiated foodstuff even in the absence of a non-irradiated
(control) sample.
Photo-stimulated luminescence (PSL) is based on the emission of
trapped energy as light that may be induced photochemically. Mineral debris,
which typically comprises silicates or bioinorganic materials such as calcite,
quartz and feldspars, can be found in most food, and these materials store
energy when they are exposed to ionizing radiation (Zhang et al. 2013). These
poly-minerals when stimulated using light source, the stored energy is
released. Dose dependency for the PSL signals is not always apparent, most
209
probably due to variation of mineral grains accidentally being on the surface of
the sample layer (Bayram & Delincee 2004). The PSL technique has been
validated by the European Committee for Standardization (EN 13571, 2009) for
food such as spices and aromatic herbs. In view of the increasing use of
irradiation technology for decontamination of herbs, this study aims to
investigate whether PSL measurement can be employed to identify three
different types of Malaysian herbs.
The objective of this study is to establish baseline data on PSL
measurement of locally available herbs for identification of γ-ray irradiation
treatment.
Three types of herbs, namely mas cotek (Ficus deltoidea), misai kucing
(Orthosiphon stamineus) and hempedu bumi (Andrographis paniculata) were
purchased from local market and packed in black polyethylene bags (about 10
g each bag). The samples were exposed to γ-radiation at the dose of 5 kGy at
room temperature using a cobalt-60 gamma source (Gamma cell 220, at
Malaysian Nuclear Agency).
PSL measurements were performed as described by the European
Standard (EN 13751: 2009) using a SURRC PPSL Irradiated Food Screening
System (SURRC: Scottish Universities Research and Reactor Center).
Approximately 2 g of samples were placed in a disposable 50 mm diameter
petri dish (Bibby sterlin type 122, Glasgow, UK) and the PSL photon counts
(PCs) were measured in subdued light condition. The samples were classified as
negative (non-irradiated) if the PSL counts was less than the lower threshold
(T1) value of 700 PCs/ 60 s while PSL counts above upper threshold (T2) value
of 5000 PCs/60 s indicated positive (irradiated) samples. Samples having PSL
signal between the two values were reported as intermediate and require
further testing for confirmation. All measurements were done in triplicates
(n=3) and the results were reported as means ± standard deviation.
According to EN 13751:2009, calibrated PSL measurements are
recommended to estimate sample PSL sensitivity, in case of ambiguous results
of an irradiation treatment or for herbs having low mineral contents.
Calibrated PSL was performed by subjecting all the herbs to 1 kGy radiation
dose after the initial PSL measurement, and then re-measured. Sensitivity
index (ratio of calibrated to initial PSL signal) was determined for the irradiated
herbs.
210
The PSL photon counts (PCs) for all samples, measured as a function of
irradiation dose are presented in Figure 1. The PCs of all non-irradiated
samples were less than the lower threshold value (700 counts/60 s), indicating
them as negative (T1, non-irradiated). The PCs of the 5 kGy irradiated herbs
measured
were
higher
than
the
upper
threshold
value
(T2 = 5000 counts/60 s) indicating irradiation treatment. Among the three
herbs, mas cotek recorded the highest PCs with values over three times more
compared to misai kucing and hempedu bumi.
Figure 1. Effect of irradiation dose on the signal intensity of mas cotek, misai
and hempedu bumi
The PSL technique, recognized as a screening tool to identify irradiated
food, has been reported to give false positive and negative results in special
cases. Nevertheless, PSL has been successfully employed to a broad range of
food for screening of the irradiated samples (Sanderson et al. 1996).
The index sensitivity of the irradiated herbs is shown in Table 1.
According to Liwen et al. (2013), index sensitivity value <10 indicates irradiated
whilst values >10 showed non-irradiated samples. All non-irradiated herbs
produced index sensitivity of >10, with values ranging from 14 to 152 for misai
kucing and mas cotek, respectively. Differences in the index sensitivity values
of non-irradiated herbs were likely due to the variation in the types and
211
quantity of silicate materials such as feldspar and quartz contaminating the
herbs, as reported by Bayram & Delincée (2004). Based on the results obtained,
it was clear that index sensitivity value is a better indicator than using the PSL
response to discriminate between non-irradiated and irradiated samples.
Table 1. Index sensitivity of irradiated herbs
Herbs
Radiation Dose (kGy)
Mas cotek
Control
5
Misai kucing
Control
5
Hempedu bumi
Control
5
*Calibrated PSL/initial PSL
Index Sensitivity*
152
0.8
14
0.8
16
0.8
CONCLUSION
Photo-stimulated luminesens (PSL) can be used to detect the irradiation
treatment of the three types of herbs. All irradiated samples were successfully
distinguished from the non-irradiated samples. The results provide a useful
database on the applicability of PSL technique in detection of irradiated herbs.
The availability of the method to detect irradiated food in Malaysia will be
useful to the Ministry of Health for enforcing labeling control in Malaysia.
ACKNOWLEDGEMENTS
The authors wish to express their gratitude to the Malaysian Nuclear Agency
(MOSTI) for their support in this project.
REFERENCES
Akram, K., Ahn, J. & Kwon, J. (2012). Chapter 1. Analytical Methods for the
Identification of Irradiated Foods. Pp. 1─36 in Belotserkovsky, E. &
Ostaltsov, Z. (eds.) Ionizing Radiation.
Bayram, G. & Delincée, H. (2004). Identification of Irradiated Turkish Foodstuffs
Combining Various Physical Detection Methods. Food Control 15: 81–
91.
212
European Standard EN 13751, (2009). Foodstuff - Detection of Irradiated Food
Using Photostimulated Luminescenc. Brussels, Belgium: European
Committee for Standardization.
Sanderson, D.C.W., Carmichael, L.A., Spencer, J.Q. & Naylor, J.D. (1996). Recent
Advances in Thermoluminescence and Photo-Stimulated Luminescence
Detection Methods for Irradiated Foods, International Meeting on
Analytical Detection Methods for Irradiation Treatment of Foods. Pp.
124–38 in McMurray, C.H., Stewart, E.M., Gray, R. & Pearce, J. (eds.).
Detection Methods for Irradiated Food—Current Status. Cambridge,
U.K.: Royal Society of Chemistry.
Zhang, L., Lin, T., Jiang, Y. & Bi F. (2013). A New Criterion of Photostimulated
Luminescence (PSL) Method to Detect Irradiated Traditional Chinese
Medicinal Herbs. Radiation Physics and Chemistry 92: 105–111.
213
OPTIMIZATION OF ANTIOXIDANT ACTIVITY IN CLINACANTHUS NUTANS
(BELALAI GAJAH/SABAH SNAKE GRASS)
HS Kong, KH Musa & NA Sani
Food Science Programme, School of Chemical Sciences and Food Technology,
Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600
UKM, Bangi Selangor, Malaysia
Tel: 03-8921 4053
Fax: 03-8921 3232
ABSTRACT
This study focused on the effects of solvents, extraction time and repeated extraction
towards antioxidant properties of Clinacanthus nutans (Belalai Gajah/Sabah Snake
Grass). Freeze-dried stems were homogenized before testing its antioxidant activity:
folin–ciocalteu index (TPC), ferric reducing/antioxidant power (FRAP) and radicalscavenging activity (DPPH) on the effect of different solvents (absolute methanol,
ethanol, acetone, and their aqueous solutions at 50%, 70%, and 100% concentrations),
st
nd
rd
time of extraction (15, 30 and 60 min) and repeated extraction (1 , 2 and 3 times).
The data obtained were statistically analyzed using SPSS Version 22 (Chicago, Inc.).
st
High antioxidant activity was shown on the 1 extraction session of 0.1 g of sample
immersed with 10 ml of 70% acetone in 60 minutes extraction time. In conclusion,
high antioxidant activity was observed in C. nutans, which can be contributed as
natural antioxidant source for human consumption. More optimization options on C.
nutans can be conducted to cultivate more nutrients from the medicinal plant and thus
contributing for human health.
Keywords: antioxidant, belalai gajah, Clinacanthus nutans, medicinal plant,
sabah snake grass
INTRODUCTION
Clinacanthus nutans is a native medicinal herb that grows in tropical climate,
mainly found in Malaysia and Thailand. C. nutans has been utilized for its
benefits and functions according to folklore, especially in the Southeast Asia
region. C. nutans is used to boost immunity, detoxification, promotes general
health condition, promotes bowel movement, as skin care, and diuresis. It is
also known to treat cancer of the lung, ovary, uterine, prostate,
nasopharyngeal and breast cancer. Most of the time, it is used for home
decoration, tea and bath (Siew et al. 2014). C. nutans is also used to treat skin
affections, insect and snakebites and swellings due to fall or boils
(Chiwapreecha et al. 2014). It is used to treat dysentery, diabetes, dysuria and
kidney disease in Indonesia, while it boosts the immune system, detoxifies,
214
promotes good health, prevents breast cancer, promote bowel movement, skin
care and goiter in Singapore (Lau et al. 2014). However, there is not much
study on its antioxidant activities. This study is focused on the effects of
solvents, extraction time and repeated extractions toward antioxidant
properties of C. nutans.
Freeze-dried stems were homogenized into 0.5 mm size before extraction. The
extraction procedure was conducted with (0.1 g) dried samples and 10 mL
extraction solvent for overnight. Solvents systems used were absolute
methanol, ethanol, acetone (Merck, Germany), and their aqueous solutions at
50%, 70%, and 100% concentrations (Musa et al. 2011). All extracted samples
were centrifuged using tabletop centrifuge (Kubota, Japan) for 10 min at
2,580×g. The supernatants were collected for further analysis. In the second
part of this study, extraction time (15, 30 and 60 min) and extraction sessions
(1st, 2nd and 3rd times) were studied using the best solvent selected in the first
part of the study. All tests such as Folin–Ciocalteu Index (Slinkard & Singleton
1977), Ferric Reducing/Antioxidant Power (Benzie & Strain 1996) and RadicalScavenging Activity (Musa et al. 2011) were performed at room temperature.
All the experiments were done in triplicates. The data obtained were
statistically analyzed using SPSS Version 22 (Chicago, Inc.) using one-way
ANOVA followed by Duncan’s Multiple Range Test.
Strong antioxidant activity was shown by solvent 70% acetone in TPC (177.69
mg of gallic acid equivalent per g of dried sample) and DPPH (231.87 mg of
trolox equivalent per g of dried sample), which was similar to the antioxidant
activity observed in pink-flesh guava of Musa et al. (2011), in a similar method.
For the effect of extraction time on antioxidant study, extraction time of 60
min exhibited the highest (p<0.05) antioxidant activities in TPC and FRAP
(221.64 mg gallic acid equivalent per g of dried sample in TPC, 3.26 mg of trolox
equivalent per g of dried sample in FRAP and 2.38 mg of trolox equivalent per g
of dried sample in DPPH). The TPC results of Lusia et al. (2015) stated 177.80 ±
19.10 mg TAE/ L for C. nutans tea, which is observed in 20 min infusion of
unfermented microwave-oven dried leaves. The results were much lower than
the current results of 15 min (886.84 mg gallic acid equivalent per g of dried
sample in TPC). There was an ascending trend on the antioxidant activities
when the extraction time was prolonged. The first extraction showed the
highest (p<0.05) antioxidant activity than the second and third extraction
215
(207.59 mg gallic acid equivalent per g of dried sample in TPC, 2.76 mg of trolox
equivalent per g of dried sample in FRAP and 2.69 mg of trolox equivalent per g
of dried sample in DPPH). More optimization options on C. nutans can be
conducted to cultivate more nutrients from the medicinal plant and thus
contributing towards human health. Future studies will focus on antimicrobial
activity, nutritive value and product development on C. nutans.
CONCLUSION
st
High antioxidant activity was shown on the 1 extraction session of 0.1 g of sample
immersed with 10 ml of 70% acetone in 60 min extraction time. Since high antioxidant
activity was observed in C. nutans, thus it can function as a good natural antioxidant
source for human consumption.
ACKNOWLEDGEMENT
The authors would like to thank Universiti Kebangsaan Malaysia for providing
the postgraduate scholarship (Zamalah), food antioxidant research lab facilities
and the following project funds: FRGS/1/2014/STWN03/UKM/02/1, GSP-2013019 and STGL-007-2008.
REFERENCES
Benzie, I. F. & Strain, J. J. (1996). The Ferric Reducing Ability of Plasma (FRAP) as a
Measure of Antioxidant Power the FRAP Assay. Analytic Biochemistry 239:
70─76.
Chiwapreecha, B., Janprasert, K. & Kongpakdee, C. (2014). Comparative Anatomy of
Three Medicinal Plants in Acanthaceae. Acta Horticulturae 1023: 229─232.
Lau, K. W., Lee, S. K. & Chin, J. H. (2014). Effect of the Methanol Leaves Extract Of
Clinacanthus nutans on the Activity of Acetylcholinesterase In Male Mice.
Journal of Acute Disease: 3.
Lusia, B. M., Hasmadi, M., Zaleha, A. Z. & Mohd Fadzelly, A. B. (2015). Effect of
Different Drying Methods on Phytochemicals and Antioxidant Properties of
Unfermented and Fermented Teas From Sabah Snake Grass (Clinacanthus
nutans Lind.) Leaves. International Food Research Journal 22(2): 661─670.
Musa, K. H., Abdullah, A., Jusoh, K. & Subramaniam, V. (2011). Antioxidant Activity of
Pink-Flesh Guava (Psidium guajava L.): Effect of Extraction Techniques and
Solvents. Food Analysis Methods 4(2011): 100–107.
216
Siew, Y. Y., Zareisedehizadeh, S., Seetoh, W. G., Neo, S. Y., Tan, C. H. & Koh, H. L.
(2014). Ethnobotanical Survey of Usage of Fresh Medicinal Plants in
Singapore. Journal of Ethnopharmacology 155(3): 1450─1466.
Slinkard, K. & Singleton, V. (1977). Total Phenol Analysis; Automation and Comparison
with Manual Methods. American Journal of Enology and Vitivulture
28:49─55.
217
EFFECTS OF EQUAL DOSES OF SULFORAPHANE, CURCUMIN AND
QUERCETIN ON HEME OXYGENASE 1 GENE EXPRESSION IN MICE LIVER
A Abdullah, N Alrawaiq & A Elbadri
Department of Pharmacology, Faculty of Medicine, Universiti Kebangsaan
Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras,
Kuala Lumpur, Malaysia
ABSTRACT
Heme oxygenase-1 (HO-1) is an enzyme that possesses anti-oxidant, antiinflammatory and cytoprotective functions. Induction of HO-1 occurs as an
adaptive and beneficial response to various injurious stimuli such as oxidative
stress. This study is aimed at monitoring the effects of equal doses (50 mg/kg
body weight) of sulforaphane, curcumin and quercetin on the expression levels
of HO-1 gene. Twelve ICR male white mice (25–30 g) were divided into 4
groups: (1) sulforaphane treated group (n=3), (2) quercetin treated group
(n=3), (3) curcumin treated group (n=3) and (4) control (vehicle) group (n=3).
The chemicals were administered intraperitoneally at a dose of 50 mg/kg body
weight for 14 days. A vehicle (DMSO, tween 20 and normal saline in the ratio of
0.05:0.1:0.85) was similarly administered to the control group. At day 15,
animals were sacrificed and their livers isolated. Total RNA was extracted,
reverse transcribed and subjected to quantitative real‐time PCR to detect HO-1
expression. Agarose gel electrophoresis was also performed to verify the
specificity of the amplification. The expression of HO-1 showed an increase of
4.6-fold following treatment with sulforaphane compared to controls (P<0.05).
Treatments with curcumin and quercetin resulted in a 3.3-fold and 3.0-fold
induction of HO-1, respectively, compared to controls (P<0.05). At a dose of 50
mg/kg body weight, sulforaphane has the highest impact on the induction of
HO-1 expression in the livers of mice, followed by curcumin and quercetin.
Keywords: Heme oxygenase-1, gene expression, sulforaphane, quercetin,
curcumin
INTRODUCTION
It is established that a diet rich in fruits and vegetables provides protection
against cancer (Steinmetz & Potter 1996). They contain antioxidants which
promote the removal of reactive species generated during normal oxidative
metabolism and unwanted xenobiotic chemicals (Ames et al. 1987). Some
218
chemicals found in food and phenolic antioxidants are chemoprotective i.e.
they induce the expression of genes and proteins involved in cellular defense.
The increased expression of cellular defense proteins provided added
protection against oxidative/chemical stress. Some of these proteins belong to
phase II drug metabolizing enzymes, although other enzymes and antioxidant
proteins were also involved. These proteins are known as phase II proteins
(Wattenberg 1978, Prestera et al. 1993, Talalay & Fahey 2001). Phase II
proteins are mainly regulated by the nuclear factor E2-related factor 2/
antioxidant response element (Nrf2/ARE) system (Kitteringham et al. 2010,
Abdullah et al. 2012). Heme oxygenase-1 (HO-1) is an example of Phase II
proteins regulated by Nrf2 (He et al. 2001). HO-1 is a rate-limiting enzyme that
catalyzes the degradation of heme (a pro-oxidant) to carbon monoxide,
biliverdin and free iron (Keum et al. 2006). HO-1 induction is important in
terms of cellular defense mechanism due to the fact that HO-1 expression is
inducible in response to various forms of cellular insult. Moreover, the end
products of HO-1 catabolism exhibit anti-oxidative, anti-inflammatory and antiapoptotic properties (Keum et al. 2006). The objective of this study is to
determine the nature and potency of HO-1 expression induced by equal doses
of several chemicals commonly found in diet i.e. sulforaphane, curcumin and
quercetin.
Twelve ICR male white mice (25–30 g) were divided into 4 groups: (1)
sulforaphane treated group (n=3), (2) quercetin treated group (n=3), (3)
curcumin treated group (n=3) and (4) control (vehicle) group (n=3). The
chemicals were administered intraperitoneally at a dose of 50 mg/kg body
weight for 14 days. A vehicle (DMSO, tween 20 and normal saline in the ratio of
0.05:0.1:0.85) was similarly administered to the control group. At day 15,
animals were sacrificed and their livers isolated. Total RNA was extracted,
reverse transcribed and subjected to quantitative real‐time PCR to detect HO-1
expression. Agarose gel electrophoresis was also performed to verify the
specificity of the amplification.
The expression of HO-1 showed an increase of 4.6-fold following treatment
with sulforaphane compared to controls (P<0.05). Treatments with curcumin
and quercetin resulted in a 3.3-fold and 3.0-fold induction of HO-1 compared
to controls (P<0.05) (Figure 1).
219
Figure 1. Effects of intraperitoneal administration of 50mg/kg sulforaphane,
curcumin and quercetin for 14 days on HO-1 gene expression in the livers of
mice. Data is presented as mean ± SEM. VH1: vehicle control group, SUL:
sulforaphane group, CUR: curcumin group, QRC: quercetin group. Amplified
products were visualized by agarose gel electrophoresis and gene expression
was confirmed by identification of the appropriate bands. GAPDH served as a
reference gene. * P < 0.05 compared to controls.
HO-1 is transcriptionally upregulated by a large variety of stimuli, including
heme, oxidative stress, signaling proteins and organic chemicals. Many of the
classical HO-1 triggers have been shown to induce HO-1 expression through
nuclear factor E2-related factor 2 (Nrf2) binding to the antioxidant response
element (ARE) at the hemeoxygenase-1 promoter region (Martin et al. 2004).
Experiments conducted on Nrf2-deficient mice showed the importance of Nrf2
in stress-dependent induction of HO-1 because HO-1 was found to be less
inducible in such mice (Cho et al. 2002). Phytochemicals such as sulforaphane,
curcumin and quercetin have been shown to induce HO-1 expression in liver
cells (hepatocytes) as well as in the liver itself (Yao et al. 2007, Bao et al. 2010
& Noh et al. 2015). However, what is not known is which of these
phytochemicals are the most potent in inducing HO-1 expression. The results of
our study clearly showed that sulforaphane is the most potent in terms of
inducing HO-1 expression in mice liver, followed by curcumin and quercetin.
Therefore, increased consumption of sulforaphane rich food such as broccoli in
humans could be beneficial in terms of general health and cancer
chemoprevention strategy.
220
CONCLUSION
Our findings indicate that at a dose of 50 mg/kg, sulforaphane has the greatest
effect in inducing HO-1 expression in mouse liver, followed by curcumin and
quercetin.
ACKNOWLEDGEMENTS
This research was funded by the Ministry of Education Malaysia & UKM, grant
code FRGS/1/2012/SKK03/UKM/02/2.
REFERENCES
Abdullah, A., Kitteringham, N.R., Jenkins, R.E, Goldring, C., Higgins, L.,
Yamamoto, M., Hayes, J. & Park, B.K. (2012). Analysis of the role of Nrf2
in the expression of liver proteins in mice using two-dimensional gelbased proteomics. Pharmacological Reports: 680─697.
Bao, W., Li, K., Rong, S., Yao, P., Hao, L., Ying, C., Zhang, X., Nussler, A. & Liu, L.
(2010). Curcumin alleviates ethanol-induced hepatocytes oxidative
damage involving heme oxygenase-1 induction. Journal of
Ethnopharmacology: 549─553.
Cho, H.Y., Jedlicka, A.E., Reddy, S.P., Kensler, T.W., Yamamoto M., Zhang, L.Y. &
Kleeberger, S.R. (2002). Role of NRF2 in protection against hyperoxic
lung injury in mice. American Journal of Respiratory Cell and Molecular
Biology: 175─182.
He, C., Gong, P., Hu, B., Stewart, D., Choi, M., Choi, A. & Alam, J. (2001)
Identification of activating transcription factor 4 (ATF4) as an Nrf2interacting protein. Implication for heme oxygenase-1 gene regulation.
Journal of Biological Chemistry: 20858─20865.
Keum, Y.-S., Han, Y.-H., Liew, C., Kim, J.-H., Xu, C., Yuan, X., Shakarjian, M. P.,
Chong, S., & Kong, A.-N. (2006). Induction of heme oxygenase-1 (HO-1)
and NAD [P] H: quinone oxidoreductase 1 (NQO1) by a phenolic
antioxidant, butylated hydroxyanisole (BHA) and its metabolite, tertbutylhydroquinone (tBHQ) in primary-cultured human and rat
hepatocytes. Pharmaceutical research: 2586─2594
221
Kitteringham, N.R., Abdullah, A., Walsh, J., Randle, L., Jenkins, R.E., Sison, R.,
Goldring, C.E., Powell, H., Sanderson, C., Williams, S., Higgins, L.,
Yamamoto, M., Hayes, J. & Park, B.K. (2010). Proteomic analysis of Nrf2
deficient transgenic mice reveals cellular defence and lipid metabolism
as primary Nrf2-dependent pathways in the liver. Journal of Proteomics.
Jun 16; 73(8): 1612–1631.
Noh, J.R., Kim, Y.H., Hwang, J.H., Choi, D.H., Kim, K.S., Oh, W.K. & Lee, C.H.
(2015).
Sulforaphane protects
against
acetaminophen-induced
hepatotoxicity. Food and Chemical Toxicology: 193─200.
Prestera, T., Zhang, Y., Spencer, S., Wilczak, C. & Talalay, P. (1993) The
electrophile counterattack response: protection against neoplasia and
toxicity. Advances in Enzyme Regulation: 33: 281─296.
Steinmetz, K.A. & Potter, J.D. (1996). Vegetables, fruit, and cancer prevention:
a review. Journal of the American Dietetic Association: 1027─1039.
Talalay, P. & Fahey, J. (2001). Phytochemicals from cruciferous plants protect
against cancer by modulating carcinogen metabolism. Journal of
Nutrition: 3027S─3033S.
Wattenberg, L.W. (1978) Inhibitors of chemical carcinogenesis. Advances in
Cancer Research: 197─226.
Yao, P., Nussler, A., Liu, L., Hao, L., Song, F., Schirmeier, A. & Nussler, N. (2007).
Quercetin protects human hepatocytes from ethanol-derived oxidative
stress by inducing heme oxygenase-1 via the MAPK/Nrf2 pathways.
Jounal of Hepatology: 253─261.
222
EVALUATION OF ANTIMICROBIAL AND ANTIOXIDANT PROPERTIES OF
ANETHUM GRAVEOLENS LEAF EXTRACTS
MN Mohd Effendi1 & AS Norrakiah2
1
Food Science Technology Research Center, MARDI Headquarters, 43400
Serdang, Selangor, 2Food Science Department, Faculty of Science and
Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor
ABSTRACT
This research was conducted to determine the antimicrobial and antioxidant
activities of dill leaves and to identify and differentiate the effects of different
solvents on dill extracts. Three solvents were used for extraction of dill leaves,
i.e. water, methanol and ethanol. Antimicrobial activity was determined by disc
diffusion test (formation of inhibition zone) and direct inhibition test (extracts
were added into growth media). In disc diffusion test, dill water extract
inhibited Bacillus cereus and Staphylococcus aureus while dill methanolic and
ethanolic extracts inhibited Escherichia coli, B. cereus, S. aureus and
Saccharomyces cerevisiae. In direct inhibition test, methanolic and ethanolic
dill extracts reduced growth of E. coli, B. cereus, and S. aureus while the water
extract reduced B. cereus and S. aureus. Dill ethanolic extract exhibited the
best antimicrobial activity compared to the other extracts. Antioxidant activity
was determined by ferric reducing antioxidant power (FRAP), ferric thiosianate
test (FTC) and thiobarbituric acid test (TBA). Dill ethanolic extract showed the
best antioxidant activity in all assays, followed by methanolic and water
extracts.
Keywords: dill, Anethum graveolens, herb, antimicrobial, antioxidant
INTRODUCTION
Anethum graveolens, commonly known as dill is a species from the family
Apiaceae or Umbelliferae. It is native to Mediterranean countries and southeastern Europe. In middle-east countries like Iran and Turkey, this herb has
been used as additive in meals because of its aroma. Dill leaves and seeds
possess essential oil with various phenolic compounds like carvone,
furanocoumarine, limonene, xanthon, triterpene and flavonoid. Singh et al.
(2005) reported that dill essential oil showed the presence of 35 phenolic
compounds and the extract, 25 compounds. The major compound was carvone
(55.2%) followed by dill apiole (43.2%), linoleic acid (23.1%), limonene (16.6%)
223
and trans-anethole (11.0%). These phenolic compounds contribute to its
antioxidant and antimicrobial properties (Cao & Prior 1998; Koleva et al. 2001).
Also, a report by Singh et al. (2005) showed that dill extract retarded the
growth of Bacillus cereus, Bacillus subtilis and Staphylococcus aureus and
fungus Aspergillus niger, Fusarium gramenearum and Penicillium viridicatum.
It was also shown that dill possessed greater antioxidant activities compared to
synthetic antioxidant, BHA and BHT. The objective of this project is to study
and compare antioxidant and antimicrobial activities of various dill extracts.
Three different solvents were used to extract dill leaves, i.e. ethanol, methanol
and water.
Leaf Extraction
Dill leaves were washed and dried in oven at 50˚C overnight. Dried samples
were grinded and weighed, and soaked in ethanol (99.9%) at ratio 1:6
(sample:ethanol) (w:v) for 3 days. The extract was filtrated (Whatman No. 1)
using Buchner flask in vacuum condition. The extract was then concentrated
using rotary evaporator (Buchi R-200) at 55 ˚C. The obtained dried extract was
weighed and kept at 4˚C for testing. Extraction of dill leaves using methanol
was similar to ethanolic extraction. Methanol (99.7%) was used to replace
ethanol. Meanwhile, for water extraction, fresh dill leaves were soaked in
distilled water at ratio 1:3 (sample:water) (w:v) and grinded. Extract was
filtrated (Whatman No. 1) using Buchner flask in vacuum. Filtrate was then
concentrated using rotary evaporator (Buchi R-200) at 100 ˚C. All extracts were
then dissolved in distilled water to produce extract solutions with different
concentrations (200, 400, 600, 800, 1000 ppm).
Analysis of Antimicrobial and Antioxidant Activities
Two methods were used to determine the antimicrobial activities of dill leaf
extracts, namely disc diffusion test (Gulcin 2003) and direct inhibition test
(Ramdas 1998) with slight modification. Meanwhile, to investigate the
antioxidant activities, three tests were used, i.e. ferric reducing antioxidant
power (Oyaizu 1986), ferric thiocyanate test (FTC) by Osawa and Namiki (1981)
and thiobarbituric acid test (TBA) by Kikuzaki and Nakatani (1993).
224
Extraction Yield
Dill water extract showed the highest extraction yield (4.17%), followed by
methanol extract (2.53%) and ethanol extract (2.47%). All three extracts were
dark green in colour with strong aroma.
Disc Diffusion Test
In the disc diffusion test, the dill water extract inhibited Bacillus cereus and
Staphylococcus aureus, while dill methanolic and ethanolic extracts were found
to be effective against Escherichia coli, Bacillus cereus, Staphylococcus aureus
and Saccharomyces cerevisiae. As the concentration of the extract increased,
the inhibition zone diameter increased. Singh et al. (2005) reported that in agar
well diffusion method, dill extract showed better results in comparison to
commercial bactericide (ampicillin). It was found effective towards Bacillus
cereus, Staphylococcus aureus and Bacillus subtilis, but less effective towards
Escherichia coli, Pseudomonas aeruginosa and Salmonella typhi. Carvone, the
major component of dill essential oil has been shown to inhibit the growth of
bacteria (Agrawal et al. 2002) and some fungus (Smid et al. 1995).
Direct Inhibition Test
This method is also known as poison food technique, where the extract was
added into bacteria media growth. The dill ethanolic and methanolic extracts
inhibited E. coli, B. cereus and S. aureus whereas water extract inhibited B.
cereus and S. aureus. Reduction percentage of bacteria growth increased as the
concentration increased. At concentration 1,000 ppm, ethanolic extract
showed the highest percentage of growth inhibition (43.48% against E. coli,
88.23% against B. cereus, 91.81% against S. aureus) compared to other extracts
against these three bacteria (p<0.05). Using the same method, dill extract was
reported to be effective towards Penicillium citrinum, Aspergillus niger,
Fusarium gramenearum, Aspergillus ochraceus, Penicillium viridicatum (Singh
et al. 2005) and Colletotrichum lindemuthianum (Shridhar et al. 2003).
Ferric reducing antioxidant power
The reducing power of three dill extracts along with BHA and BHT increased
with concentration. Ethanolic extract showed the highest absorbance reading
compared to methanolic and water extracts, and positive control (p<0.05). At
1000 ppm, the sequence for reducing power was ethanolic extract >
methanolic extract > water extract > BHA > BHT. Singh et al. (2005) reported
225
that dill extract reduced ferum (III) better than synthetic antioxidant BHA, BHT
and propyl galate.
Ferric thiocianate test
This method was used to evaluate the peroxide level during initial stage of
peroxidation. Low absorbance values would indicate high level of antioxidative
activity (Singh et al. 2005). After seven days, all extracts effectively inhibited
linoleic acid oxidation. Ethanolic and Methanolic extracts showed the best
results compared to water extract, BHA, BHT and control. Singh et al. (2005)
reported that Anethum graveolens extract effectively inhibited linoleic acid
oxidation compared to synthetic antioxidant.
Thiobarbituric acid test
This method was used to measure the secondary product of oxidation such as
aldehyde and ketone. Ethanolic and methanolic extracts showed the lowest
absorbance values at day 5 to 7 compared to water extract, BHA ,BHT and
control. This indicated that the amount of peroxidation was lower in ethanolic
and methanolic extracts until day 7, compared to other extracts.
Type of solvent used in extracting herbs contributed towards its
antioxidant activity. Herbs extracted with non-polar solvents show better
antioxidant activity than polar solvents (Pinelo et al. 2004). Different active
compounds with different polarity are present in extracts in various amounts.
A study by Shyu (2009) showed that different solvents extracted different
phenolic compounds. The highest total amounts of polyphenols, flavonoids,
anthocyanins and proanthocyanidins were found in the ethyl acetate extract,
whereas the highest total amounts of ascorbic acid and tocopherols were
found in ethanolic extract. The study also demonstrated that ethyl acetate
extract had the highest antioxidant propeties, followed by ethanolic and nhexane extracts.
CONCLUSION
It can be concluded that all three extracts of dill (A. graveolens) showed good
antimicrobial and antioxidant properties. However, dill extracted with ethanol
and methanol showed better antimicrobial and antioxidant activitie compared
to the water extract.
226
ACKNOWLEDGEMENT
The author would like to thank Food Science Department, Faculty of Sciences
and Technology, Universiti Kebangsaan Malaysia (UKM) for funding and
technical support.
REFERENCES
Agrawal, K.K., Khanuja, S.P.S., Ahmed, A., Santhakumar, T.R., Gupta, V.K. &
Kumar, S. (2002). Antimcrobial Activity Profiles of the Two Enantiomers
of Limonene and Carvone Isolated from Oils of Mentha spicata and
Anethum sowa. Flavour Fragrance Journal 17: 59─63.
Gulcin, I., Oktay, M., Kirecci, E. & Kufrevioglu, I. (2003). Screening of
Antioxidant and Antimicrobial Activities of Anise (Pimpinella anisum L.)
Seed Extracts. Food Chem. 83: 371─382.
Osawa, T. & Namaki, M. (1983). A Novel Type Antioxidant Isolated from Leaf
Wax of Eucalyptus Leaves. Agric Biol Chem. 45:735─739.
Oyaizu, M. (1986). Studies on Products of Browning Reactions: Antioxidant
Activities of the Products of Browning Reaction Prepared from
Glucosamine. Japanese Jounal of Nutr. 44: 307─315.
Kikuzaki, H. & Natakani, N. (1993). Antioxidant Effect of Some Ginger
Constituents. Journal of Food Sci. 578: 1407─1410.
Shyu, Y.S., Lin, Y.S., Chang, Y.T., Chiang, C.J & Yang, D.J. (2009). Evaluation of
Antioxidant Ability Extract from Dill (Anethum graveolens L.) Flower.
Food Chemistry 115: 515─521.
Singh, G., Maurya S., de Lampasona, M.P & Catalan, C. (2005). Chemical
Constituents, Antimicrobial Investigations and Antioxidative Potentials
of Anethum graveolens L. Essential Oil and Acetone Extract: Part 52.
Journal of Food Sc. 70: 208─215.
Smid, E.J., de Witte, Y. & Gorris, L.G.M. (1995). Secondary Plant Metabolites as
Control Agents of Post Harvest Penicillium Rot on Tulip Bulbs.
Postharvest Biotechnology 6: 303─312.
227
ASAM GELUGUR POWDER RICH IN HCA (HYDROXYCITRIC ACID): A
POTENTIAL CROP FOR WEIGHT MANAGEMENT
HZ Umi Kalsum, HA Hashimah, A Sharizan, A Mohamed Nazim, M Aida, S Nor
Fadhilah & A Mohd. Fadli
Food Science Technology Research Centre, MARDI Headquarters, Persiaran
MARDI-UPM, 43400 Serdang, Selangor, Malaysia
Tel: 03-89536397
Fax: 03-89536500
ABSTRACT
Garcinia atroviridis (asam gelugur) is one of the underutilised plants used
frequently in medicine as fruit for reducing weight and excess body fat by
halting the glycogen process. The principal acid in this fruit has been found to
be (-)-hydroxycitric acid (HCA). This study aimed to optimize the HCA content in
the powder product (G. atroviridis) which will be used as ingredient that can be
incorporated to a variety of health food products that act as agents for weight
management. Processing parameters of asam gelugur powder had been
produced. Pretreatment of enzymes and blanching process of 3 and 4 months
old asam gelugur fruit mixture was used in this activity. Determination of
enzyme and time blanching percentage determined by Respond Surface
Methodology using Composite Design Centre. A total of 13 run has been
determined. From the statistic analysis, it showed that the percentage of
enzyme has a significant effect (p<0.05) on HCA content. The blanching
duration also has a significant effect (p<0.05) on HCA content. To conclude,
these two pre-treatments gave positive effect on the increment of HCA content
in asam gelugur powder.
Keywords: Garcinia atroviridis, hydroxycitric acid, weight management
INTRODUCTION
Garcinia atroviridis, also known as asam gelugur, asam gelugo, or asam keping
in Malay is a medium-sized fruit of a large rainforest tree native to Peninsular
Malaysia. This species grows wild throughout Peninsular Malaysia but is also
widely cultivated, especially in the northern states, owing to its economic and
medicinal value. Sun-dried slices of the fruits are commercially available and
popularly used as a seasoning in curries, sour relish and also for dressing fish
(Corner 1988). They also have been used to improve blood circulation, treat
coughs and as an expectorant and laxative (Amran et al. 2009). G. atroviridis is
228
one of the underutilised plants used frequently in medicine as fruit for
reducing weight and excess body fat by halting the glycogen process.
Obesity is a common chronic disorder of carbohydrate and fat
metabolism which is characterized by excessive fat deposition in adipose tissue
and other internal organs such as liver, heart, pancreatic islet and skeletal
muscle (World Health Organization 1997; Ahima 2006). It remains a major
global public health issue because of its increasing prevalence, cutting across
all sex, age-groups, ethnicity or race (World Health Organization 1998). Rapid
weight loss products and programs dominate the focused marketers and
consumers alike. Furthermore, rapid weight loss is potentially unhealthy and
disturbs metabolic set-point homeostasis, including undesirable rebound
weight gain consequences (Bernad et al. 2005). HCA has been reported to
safely promote weight loss in laboratory animals and humans without
stimulating the central nervous system (Ohia et al. 2002; Preuss et al. 2004;
Jena et al. 2002).
The optimization of bioactive compound i.e. L-Hydroxycitric acid (HCA)
may be achieved for the production of functional foods with enhanced
functionality and stability. Therefore, this study was carried out to determine
the pre-treatment optimization of HCA in G. atroviridis for the production of
high HCA ingredient that will be incorporated into any weight management
products.
Optimization of Pre-Treatment
Processing parameters of asam gelugur powder had been established.
Pretreatment of enzymes and blanching process of 3 and 4 months old asam
gelugur fruit mixture had been used in this activity. Determination of enzyme
and time blanching percentage determined by the method of Respond Surface
Methodology using Composite Design Centre. A total of 13 run has been
determined.
Experimental Design and Statistical Analysis
The HCA determination was carried out in three replicates. All determinations
were statistically analyzed by the analysis of variance (ANOVA. The Duncan
Multiple Range Test was used to detect the differences between samples
(Gomez & Gomez 1984).
229
RESULTS & DISCUSSION
Optimization of Pre-Treatment
From the statistical analysis, Table 1 shows that the second order on
percentage of enzyme has a significant effect (p<0.05) on HCA content. Same
goes to boiling duration that this pre-treatment also has a significant effect
(p<0.05) on HCA content.
Table 1. Regression coefficients (R2) and p (probability values) for HCA of asam
gelugur sample
HCA
Suggested model :
quadratic
Regression coefficient
Constant,
2.96
* percentage of enzyme, b1
-3.193E-003
* boiling duration, b2
-0.13
* percentage of enzyme, b21
-0.35*
2
* boiling duration, b 2
-0.39*
* percentage of enzyme * boiling duration, b12
-0.11
R2
0.7603
P or probability
0.1133
Subscripts: 1 = ultrasound amplitude, 2 = sonication time
* Significant at 0.05 level,
** Significant at 0.01 level, *** Significant at 0.001 level
DESIGN-EXPERT Plot
HCA
X = A: Enzyme
Y = B: Blanched
2.97434
2.72452
2.4747
HCA
2.22488
1.97506
0.50
0.40
30.00
23.75
0.30
17.50
0.20
11.25
B: Blanched
5.00
230
0.10
A: Enzyme
Figure 1. HCA content in asam gelugur in comparison with commercial
products.
CONCLUSION
In conclusion, these two pre-treatments gave positive effect on the increment
of HCA content in asam gelugur powder. Asam gelugur powder HCA content
higher than that of commercial product’s has been successfully developed
(Figure1).
REFERENCES
Ahima, R.S. (2006). Obesity Epidemic in Need of Answers. Gastroenterology.
131: 991
Amran, A.A., Zaiton, Z., Faizah, O. & Morat, P. (2009). Effect of Garcinia
Atroviridis on Serum Profiles and Atherosclerotic Lesions in The Aorta of
Guinea Pigs Fed a High Cholesterol Diet. Singapore Med J. 50 (3):
295─299.
Bernad, W.D., Manashi, B., Gottumukkala, V.S., Micheal, A. S., Harry, G.P. &
Debasis, B. (2005). Bioefficacy of a Novel Calcium-Potassium Salt Of (-)Hydroxycitric Acid. Mutation Research. 579: 149─162.
Corner, E.J. (1988). Wayside Tress of Malaya. Malayan Nature Society, Kuala
Lumpur.
Gomez, K.A & Gomez, A.A. (1984). Statistical Procedures for Agricultural
Research 2nd ed., p.208─215. New York: John Wiley.
231
Jena, B.S., Jayaprakasha, G.K., Singh, R.P., & Sakariah, K.K. (2002). Chemistry
and Biochemistry of (-)-Hydroxycitric Acid from Garcinia. J.Agric. Food
Chem. 50:10─22
Ohia, S.E., Opere, C.A., Leday, M., Bagchi, D. & Bagchi S.J. (2002). Stohs, Safety
and Mechanism of Appetite Suppression by a Novel Hydroxycitric Acid
Extract (HCA-SX). Molecul Cell Biochem. 238:89─103
Preuss, H.G., Bagchi, D., Bagchi, M., Rao, C.V., Satyanayana, S. & Dey, D.K.
(2004). Efficacy of a Novel, Natural Extract Of (-)-Hydroxycitric Acid
(HCA-SX) and a Combination of HCA-SX, Niacin Bound Chromium and
Gymnema Sylvestre Extract in Weight Management in Human
Volunteers: a Pilot Study. Nutr. Res. 24: 45─58.
World Health Organization. (1997). Obesity: Preventing and Managing The
Global Epidemic. Report of a WHO Consultation. Presented at the World
Health Organization Headquarters, Geneva, Switzerland. June 3─5 1997.
World Health Organization. (1998). Obesity: Preventing and Managing The
Global Epidemic. Report of a WHO Consultation on obesity.
WHO/NUT/NCD/98. Geneva, Switzerland. pp 1─275.
232
TOXICOLOGY STUDY OF HIBISCUS SABDARIFFA L. LEAVES EXTRACT ON
NORMAL SPRAGUE-DAWLEY RATS
H Hasnisa, M Syahida, H Hadijah, Z Kharis, A Sharizan, D Mohd Nazrul
Hisham, R Suri, ZJ Arif, S Ahmad Tarmizi & MF Nurul Nabilah
Food Science Technology Research Centre, MARDI Headquarters, Persiaran
MARDI-UPM, 43400 Serdang, Selangor
Tel: 03-89536456
Fax: 03-89536500
ABSTRACT
Roselle is commonly incorporated in foods and has not previously been
evaluated for safety through standard in vivo toxicological studies. In the
present study, acute and sub-acute oral toxicity studies of roselle leaves were
designed and conducted in Sprague Dawley (SD) rats. Acute oral administration
of roselle leaves extract (RLE) was done as a single dose at 3000 mg/kg of body
weight (BW). Meanwhile, sub-acute toxicity study was done by oral
administration at doses of 0 (control), 1000 mg/kg, 2000 mg/kg and 5000
mg/kg of BW for 28 days. There were no mortality, adverse clinical signs and
abnormal changes in body weight observed within 14 days of the acute toxicity
study. In the sub-acute oral toxicity study, no mortality and toxicologically
significant changes in clinical signs, haematology, clinical biochemistry and
organ weights except for slight significant decrease on relative liver weight,
glucose and trigliseride (TG) level. These observations suggest that RLE is
practically non-toxic for SD rats in oral acute and sub-acute toxicity study and
the no-observed-adverse-effect level (NOAEL) of RLE in rats is 5000 mg/kg
when administration orally for 28 days.
Keywords: Hibiscus sabdariffa L., Sprague Dawley rats, acute, sub-acute
INTRODUCTION
Hibiscus sabdariffa Linne, also known as roselle grows in many tropical and
sub-tropical countries and is one of highest volume specialty botanical
products in international commerce (Halimatul et al. 2007). The swollen
calyces are of commercial interest and the leaves can be used for animal
fodder and fiber. In addition, roselle leaves are rich in polyphenols (which act
as antioxidants that can remove harmful free radicals) have high potential to
be used as a health food product with pharmacological activities (Qi et al.
2005). In line with efforts to balance the conservation of biodiversity and
encouraging controlled exploitation of plant resources for economic gains
especially in biopharming, roselle leaves can be converted into high value
233
products (from agricultural waste). Therefore, it is important to maximize the
usage of this beneficial component for overall health and well being. However,
there is limited information on the safety of roselle leaves and never been
scientifically evaluated through standard in vivo toxicological studies.
Therefore, there is a pressing need to clarify the toxicological profile of the
leaves. In this study, a comprehensive safety evaluation on roselle leaves
extract (RLE) was conducted by performing a single dose acute and 28 days
repeated dose of sub-acute oral toxicity studies in Sprague Dawley (SD) rats.
Sample Preparation
The roselle leaves were dried at 40 °C (moisture content < 10%) and ground.
The extract of roselle leaves was prepared by infusing the dried leaves with
boiling water for 10 min. The extract were prepared at 3000 mg/kg of body
weight (BW) for acute study and three different concentrations (1000, 2000
and 5000 mg/kg BW) for sub-acute oral toxicity study.
Acute Oral Toxicity Study
Sprague Dawley rats were randomly divided into two groups (control group
and RLE treatment group) consisting of ten animals (5 males and 5 females
weighing 150-200 g per group) in each group. After overnight fasting (8–10 h),
a single RLE dose of 3000 mg/kg BW was given to the treatment group whereas
the control rats received only distilled water. All animals were observed for
clinical signs including mortality and any adverse reactions immediately after
dosing at 1, 2, 4 and 6 h, then once daily until day 14. The body weight was
measured once before the commencement of the dosing and then daily until
day 14 (Abdullah et al. 2009).
Sub-Acute Oral Toxicity Study
Twenty female SD rats weighing 200-250 g were randomly assigned into four
groups; a control and three treatment groups (n=5). During the oral
administration period, SD rats were respectively dosed with distilled water
(control group, C) or RLE at doses of 1000 mg/kg BW (LD, low dose), 2000
mg/kg BW (MD, medium dose) and 5000 mg/kg BW (HD, high dose) of extract
via drinking bottles as they can access ad libitum. On average, each rat will get
100 ml of sample per day. Any remaining sample left will be measured. General
appearance or behaviour of each rat was observed daily during the 28-days
study and the body weight was recorded every week (Ryu et al. 2004).
234
Haematology and Serum Biochemistry Analysis
The animals were fasted for approximately 12 h and blood samples for
haematological and biochemistry analyses were withdrawn under light ether
anesthesia from posterior vena cava. An aliquot of blood per animal
(approximately 20 µL) was treated in a 3 ml ethylen-diamino-tetracetic-acid
(K3-EDTA) tube to analyze haematological indexes. The blood sample was
analysed for complete blood profile by using Haematology Analyzer (Medonic
CA530, Italy). For serum biochemical blood analysis, one aliquot of blood per
animal was placed in a 5 ml Z-serum tube and centrifuged at 3,000 rpm for 20
min. The serum was analyzed by using Blood Clinical Analyzer (Vitalab Selectra
E, Italy).
Acute Oral Toxicity Study
There were neither significant toxicity symptoms nor death occurred to the
acute experimental rats during the first 6 hours and within 14 days in acute
toxicity studies. Figure 1 and Figure 2 showed no significant differences in
body weights increment between the RLE administrated group and the control
in both sexes. This result showed that the RLE is safe since there were neither
toxic signs nor mortality observed after administration of single high dose
(3000 mg/kg BW) extract.
Figure 1. Normalized body weight of
acute study for male rats (n=5; RJ,
single high dose of RLE CJ, control
group)
Figure 2. Normalized body weight of
acute study for female rats (n=5; RB,
single high dose of RLE and CB,
control group)
235
Sub-Acute Oral Toxicity Study
In sub-acute toxicity studies, there were no mortality and significant toxicity
changes in clinical signs, haematology, clinical biochemistry and organ weight
except a significant decrease in relative liver weight, glucose levels and
triglyceride (TG). The body weight assessment (Figure 3) showed that the
treated rats have gained positive body weight, indicating good health and nontoxicity effects. In haematology parameters, Figure 4 showed that there were
no significant differences between the treatment and control groups in red
blood cell (RBC), haematocrit (HCT), platelet (PLT), white blood cell count
(WBC) and haemoglobin (Hb) and the values stayed within the normal
reference range (Petterino & Argentino-Storino 2006).
Figure 5 showed the reduction of relative liver weight (p < 0.05) probably
due to the reduced final weight of the rats and cannot be concluded as a sign
of toxicity and the results also within the normal reference range of 2.43 to
3.46% (Han et al., 2010). The enzyme serum analysis (Figure 6) also showed no
signs of toxicity due to decreased levels of ALT (alanine aminotransferase) and
ALP (alkaline phosphatase) significantly. RLE is considered non-toxic due to the
lower level of these liver marker enzymes could indicate a degree of liver
protection (Zhang et al. 1996). There were also no significant changes observed
in serum protein profile (Figure 6) including total protein, albumin and globulin
level in the treated groups as compared to control. Figure 7 did not show any
kidney failure or malfunction since there were no significant difference noted
in urea (range from 5.48 to 6.27 mmol/l) and creatinine (range from 50.17 to
54.00 µmol/l) when dosage was increased from 1000 mg/kg to 5000 mg/kg as
compared to the control. Figure 8 showed that RLE reduce the glucose and TG
(trigliseride) level significantly (p < 0.05) in all dosages (1000 to 5000
mg/kg/day) as compared to control. However, the alterations were assumed to
be toxicologically irrelevant because they were within normal physiological
ranges (Petterino & Argentino-Storino 2006; Han et al. 2010), and were not
dose-related or reflected by any changes in other related parameters.
236
Figure 3. Normalized body weight
of rats for sub-acute study (n=5; RL
low dose, RM medium dose; RH
high dose and C, control group)
Figure 4. Haematology data for
female SD rats treated orally with
roselle leaves extract (RLE) for subacute study (n-5)
Figure 5. Percentage of relative
organ weight of SD rats for subacute study
Figure 6. Blood clinical parameters
for liver status in sub-acute study
Figure 7. Blood clinical parameters
for kidney status in sub-acute study
Figure 8. Blood glucose level and
lipid profile in SD rats for sub-acute
study (TC, total cholesterol; HDL,
low density lipoprotein; LDL, low
density lipoprotein; TG, trigliseride)
237
CONCLUSION
In conclusion, the study showed RLE is considered non-toxic to the SD rats and
the NOAEL level for sub-acute test is 5000 mg/kg BW. These data provide
important reference of RLE for usage as a food supplement or in future clinical
trials as a medication.
ACKNOWLEDGEMENTS
The authors are thankful to the financial supports from MARDI and Malaysian
Ministry of Agricultural for the research development grant of P-161.
REFERENCES
Abdullah, N.R., Ismail, Z. & Ismail Z. (2009). Acute Toxicity of Orthosiphon
stamineus Benth Standardized Extract in Sprague Dawley Rats.
Phytomedicine 16: 222─226.
Han, Z.Z., Xu, H.D., Kim, K.H., Ahn, T.H., Bae, J.S., Lee, J.Y., Gil, K.H., Lee, J.Y.,
Woo, S.J., Yoo, H.J., Lee, H.K., Kim, K.H., Park, C.K., Zhang, H.S. and
Song,S.W. (2010). Reference Data of the Main Physiological Parameters
in Control Sprague-Dawley Rats from Pre-clinical Toxicity Studies.
Laboratory Animal Research 26 (2): 153─164.
Halimatul, S.M.N., Amin, I., Mohd Esa, N. Nawalyah, A.G. and Siti Muskinah, M.
(2007). Protein Quality of Roselle Seeds. ASEAN Food Journal 14 (2):
131─140.
Petterino, C. and Argentino-Storino, A. (2006). Clinical Chemistry and
Haematology Historical Data in Control Sprague-Dawley Rats from Preclinical Toxicity Studies. Experimental and Toxicology Pathology 57:
213–219.
Qi, Y., Chin, K.L., Malekian, F., Berhane, M. and Gager, J. (2005). Biological
Characteristics, Nutritional and Medicinal Value of Roselle, Hibiscus
Sabdariffa. CIRCULAR – Urban Forestry Natural Resources and
Environment No. 604.
Ryu, S.D., Park, C.S., Baek, H.M., Baek, S.H., Hwang, S.Y. and Chung, W.G.
(2004). Anti-diarrheal and Spasmolytic Activities and Acute Toxicity
238
Study of Soonkijangquebo, a Herbal Anti-diarrheal Formula. Journal of
Ethnopharmacology 91: 75─80.
Zhang, M.,Song, G. and Minuk, G.Y. (1996). Effects of Hepatic Stimulator
Substance, Herbal Medicine, Selenium/Vitamin E, and Ciprofloxacin on
Cirrhosis in the Rat. Gastroenterology 110 (4): 1150─1155.
239
CLINACANTHUS NUTANS L.: SAFETY AND TOXICITY STUDY
M Syahida1, H Hasnisa1, AG Rosnani2, Z Kharis1, DMN Hisham1, R Suri1 & SA
Tarmizi1
1
Food Science Technology Research Centre, Malaysian Agriculture Research
and Development Institute (MARDI), Selangor, Malaysia
2
Horticulture Research Centre, Malaysian Agriculture Research and
Development Institute (MARDI), Selangor, Malaysia
Tel: 03-8953 6425
Fax: 03-8953 6500
ABSTRACT
This study was aimed to evaluate the effect of Clinacanthus nutans L. extract
on Sprague Dawley rats subjected to acute (single, high dose) and sub-acute
(28-day repeated doses) test. A single dose (3 g/kg of body weight) oral
administration did not show any acute oral toxicity in rats. In sub-acute study,
C. nutans L. extract was administered at 1 g/kg body weight (low dose, LD), 2
g/kg (medium dose, MD) and 5 g/kg (high dose, HD). Distilled water was given
to the rats as control. Administration of C. nutans L. extract did not cause
negative effect in blood haematology even though a statistically significant
(p<0.05) decreased in platelet level was noted. Result from serum biochemical
test showed that the consumption of the extract did not result in liver and
kidney failure since no significant changes was observed except for alanine
phosphatase (ALP), although the results were still within the normal range.
Based on these findings, the no-observed-adverse effect of C. nutans L. extract
is 5 g/kg of body weight per day.
Keywords: Clinacanthus nutans L, acute, sub-acute, Sprague-Dawley, blood
clinical
INTRODUCTION
Clinacanthus nutans L. (CNL) is a medicinal plant that has been traditionally
used especially among Chinese and Jawa people in Malaysia for anti-cancer
and anti-diabetic, but there is still lack of scientific evidence regarding its
benefits especially from local researchers. This plant belongs to Acanthaceae
family. In Malaysia, this leafy green shrub is commonly known as Sabah snake
grass or “daun belalai gajah”. Previous reports showed that CNL has antiinflammatory, anti-hepatitis and anti-herpes activities and this was confirmed
by researchers from Thailand (Sittiso et al. 2010). The plant was reported to
240
contain lupeol and β-sitosterol, which are pharmacological active components
that showed chemopreventive properties. There is still lack of scientific
research regarding its benefit and the content of bioactive compound in CNL
and the needs to determine and quantify its chemoprevention property.
Therefore, it is hoped that this study could provide scientific evidence to prove
its safety and beneficial to health.
The safety and toxicity of CNL extract was evaluated using acute and sub-acute
test. This study was approved by MARDI Animal Ethic Committee. All animals
were housed in a controlled environment with 12 hours light per day. They
were allowed to acclimatize for seven days before beginning treatment.
The acute study was conducted with a single high dose (3 g/kg body
weight (BW)) of the extract (Ryu et al. 2004) on five male and five female
Sprague-Dawley rats (150-200 g). Distilled water was used as control. The rats
were monitored within six hours after administration and for the next 14 days
if any toxicity symptoms occured. The body weight was measured during
experimental period.
The sub-acute or 28-days repeated dose study was performed based
on Ryu et al. (2004). Five male Sprague-Dawley rats weighing 200-250 g per
group were given distilled water (control dose, C), 1 g/kg BW (low dose, LD), 2
g/kg BW (medium dose, MD) and 5 g/kg BW (high dose, HD) of extract via ad
libitum 100 ml/rat/day. On day 29, all rats were fasted from food for
12 hours before sacrificed. The blood samples collected from posterior vena
cava were then analysed for haematogy (red blood cell (RBC), white blood cell
(WBC), platelet, hematocrit (HCT) and haemoglobin) and clinical parameters
(alanine amino transferase (ALT), aspartate amino transferase (AST), alanine
phosphatase (ALP) activities, bilirubin, total protein, albumin, globulin,
albumin/globulin (A/G) ratio, urea and creatinine). Blood glucose was
determined using blood taken from the tail.
Statistical analysis was performed using ANOVA followed by Duncan
New Multiple Range Test (DMRT) in SAS System, ver. 9.0. All values are
expressed as group mean ± standard deviation (SD). The minimal level of
significance accepted was p<0.05.
241
Acute Toxicity Study
After a high, single administration of CNL extract (3.0 g/kg ), there was neither
significant toxicity symptoms nor death in the first 6 hours. The body weight
increments of rats are shown in Figure 1 and Figure 2. There was no significant
difference between groups.
Acute toxicity study shows that the CNL extract is safe since there was
neither toxic sign nor mortality observed after administration of a single high
dose (3.0 g/kg ).
Figure 1. Normalized body weight of Figure 2. Normalized body weight of
acute study for male rats (n=5; BJ- acute study for female rats (n=5; BBsingle high dose of CNL extract and CJ- single high dose of CNL extract and
CB- control group)
control group)
Sub-Acute Toxicity Study
There was no mortality or abnormalities in all treated rats with respect to hair
coat, eye colour, rashes and skin irritation of animal at any dose level tested
during the 28-days study of CNL extract.
Figure 3 shows that administration of CNL extract increased the final
body weight of all rats as compared to the initial stage. The measurement of
body weight is important because changes in body weight have been used as
an indicator of adverse effect of drugs and chemicals (Teo et al. 2002; Hilaly et
al. 2004).
242
Figure 3. Normalized body weight of rats in the sub-acute test (n=5)
The results of haematology are shown in Table 1. There was no
significant difference in all haematology parameters except for platelet level.
The platelet level decreased significantly (p<0.05) in the low and medium dose.
However, the significant difference in this result did not show any toxicityrelated changes since the platelet level were within the normal range of
458.0  109/l to 1427.0  109/l (Claudio & Alberta 2006). Besides that, no
significant difference was observed in RBC, HCT and WBC levels when
compared to the control group.
Table 1. Level of blood haematology parameter after 28-day repeated dose
Analysis
12/
RBC (10 l)
HCT (%)
9
PLT (10 /l)
9
WBC (10 /l)
HGB (g/l)
Control Dose
a
7.77±0.30
a
38.21±2.73
a
1235.44±220.75
a
4.87±1.30
ab
146.56±7.18
Low Dose
a
7.27±0.63
a
35.02±3.74
b
843.00±291.95
a
4.98±1.92
ab
143.20±15.59
Medium Dose
a
14.29±16.22
a
55.34±47.40
b
699.20±363.53
a
4.42±36.27
b
114.14±63.83
High Dose
a
8.02±0.53
a
38.30±0.83
a
1252.60±280.86
a
3.78±1.03
a
159.20±2.17
Values are means ± standard deviation (SD) for five rats in each group. Means
with the same letter in same row are not significantly different (P<0.05).
Results for serum clinical test for liver parameters are listed in Table 2.
Administration of CNL extract did not show any significant difference in all
parameters except for globulin. Results on serum protein showed significant
difference in globulin level in rats given medium dose of extract as compared
to the control. However, the difference is not associated with any liver failure
or malfunction because the globulin level was still within the normal range of
30-36 g/l (Claudio & Alberta 2006).
Urea and creatinine are parameters used to determine kidney failure.
A high level of urea and creatinine in blood serum indicates the occurrence of
severe kidney damage. The results did not show any kidney failure or
malfunction since there was no significant difference noted in urea (ranged
from 6.27 ± 1.66 to 7.73 ± 0.78 mmol/l) and creatinine (ranged from 43.49 ±
21.37 to 55.80 ± 3.96 µmol/l) (Table 3).
243
Table 2. Blood chemistry parameter for liver function after 28-day repeated
dose
Analysis
ALT (U/l)
AST (U/l)
ALP (U/l)
Bilirubin
(µmol/l)
Total Protein
(g/l)
Albumin (g/l)
Globulin (g/l)
A:G
Control Dose
a
80.78±10.85
a
96.72±17.09
ab
270.33±90.94
a
2.66±0.53
Low Dose
a
88.60±10.53
a
111.28±7.33
a
338.20±83.36
a
3.13±0.40
a
73.86±5.25
a
42.36±2.88
a
31.20±2.59
a
1.35±0.05
72.93±3.17
41.77±2.60
a
31.11±3.30
a
1.36±0.20
Medium Dose
a
73.02±39.85
a
116.00±78.95
b
205.20±102.85
a
9.20±13.87
a
55.89±31.48
a
67.32±57.55
b
23.68±10.01
a
1.85±0.78
High Dose
a
90.80±22.80
a
112.99±8.92
ab
242.60±21.84
a
2.40±0.61
a
70.67±4.77
a
a
41.16±3.47
ab
29.40±2.51
a
1.40±0.16
with the same letter in the same row are not significantly different (p<0.05).
Table 3. Blood biochemistry parameter for kidney function after 28-day
repeated dose
Control Dose
Low Dose
Medium Dose
High Dose
Analysis
a
a
a
Urea (mmol/l)
6.27±1.66
7.73±0.78
7.23±2.27
6.74±0.85a
a
a
a
Creatinine (µmol/l)
51.78±6.72 52.40±4.04 43.49±21.37
55.80±3.96a
with the same letter in the same row are not significantly different (p<0.05).
CONCLUSIONS
Administration of a single, high dosage of CNL extract in acute and different
dosage in sub-acute toxicity tests showed no toxicity symptom or death. The
no-observed-adversed-effect-level (NOAEL) of this extract is 5 g/kg body
weight per day.
ACKNOWLEDGEMENT
The authors would like to thank the Ministry of Agriculture & Agro-based
Industry of Malaysia for the Development Grant and our contract staff Miss
Nur Hafiqa & Miss Siti Nur Aslina for a great help in the project.
244
a
REFFERENCES
Claudio, P. & Alberta, A.S. (2006). Clinical Chemistry and Haematology
Historical Data in Control Sprague Dawley Rats from Pre-Clinical to
Toxicity Studies. Experimental and Toxicology Pathology 57: 213–219.
Hilaly, J.E., Israili, Z.H. & Lyoussi, B. (2004). Acute and Chronic Toxicological
Studies of Ajugaiva in Experimental Animals. J. of Ethnopharmacology
91:43–50.
Ryu, S.D., Park, C.S., Baek, H.M., Baek, S.H., Hwang, S.Y. & Chung, W.G. (2004).
Anti-Diarrheal and Spasmolytic Activities and Acute Toxicity Study of
Soonkijangquebo, A Herbal Anti-Diarrheal Formula. Journal of
Ethnopharmacology 91: 75–80.
Sittiso, S., Ekalaksananan T., Pientong C., Sakdarat S., Charoensri N. &
Kongyingyoes B. (2010). Effects of Compounds from Clinacanthus nutans
on Dengue Virus Type 2 Infection. Srinagarind Medical Journal 25
(Suppl).
Teo, S., Stirling, D., Thomas, S., Hoberman, A. & Khetani, V. (2002). A 90-Day
Oral Gavage Toxicity Study of D-Methylphenidate and D,LMethylphenidate in Sprague-Dawley Rats. Toxicology 179: 183–196.
245
IN VITRO TOXICOLOGICAL EVALUATION OF 50 METHANOL EXTRACTS
FROM TRADITIONAL MEDICINAL PLANTS USED BY THE ORANG ASLI
MGH Khoo, S Rohana, M Nik Musaadah, J Fadzureena, MA Adiana, H Nuziah,
AL Tan, Z Nurul Husna, B Intan Nurulhani, HF Lim & H Norini
Forest Research Institute Malaysia, 52109 Kepong, Selangor
Tel: 03-6279 7342
Fax: 03-6272 9805
In Collaboration with: Technical Committee of Traditional Knowledge under
MoU with FRIM & JAKOA
ABSTRACT
Plants are widely used in the health care regimen since ancient times. In recent
years, various studies have investigated the biological activity and potential
medicinal values of medicinal plants. In this study, 50 methanol extracts from
15 medicinal plants used by the Orang Asli were evaluated for their potential
toxicity. Toxicity was evaluated in vitro using Vero and WRL-68 cell lines. Out of
the 50 extracts tested, 6.0% of the extracts exhibited toxicity towards both
Vero and WRL-68 cell lines with median inhibition concentration (IC50) values
below 20 g/ml; 30.0% had IC50 values ranging between 20-100 g/ml; 24.0%
with IC50 values above 100 g/ml; and 12.0% did not negatively affect the
viability of cells more than 50% at the highest concentration (625 g/ml)
tested. The level of toxicity towards Vero and WRL-68 cell lines were different
for the remaining 28.0% of the extracts (i.e. 14 extracts). Out of these 28.0%
extracts tested towards Vero cell line, the IC50 values for 21.4% were below 20
g/ml; 42.9% had IC50 values ranging between 20-100 g/ml; 28.6%
demonstrated IC50 values in the category above 100 g/ml; and 7.1% did not
cause more than 50% of cell death at the concentrations tested. Meanwhile,
for the toxicity towards WRL-68 cells of these remaining 14 extracts, 14.3%
demonstrated IC50 values that fall under the category of less than 20 g/ml;
35.7% with IC50 values ranging between 20-100 g/ml; 42.9% with values
above 100 g/ml; and 7.1% with values of more than 625 g/ml. As the saying
“The dose makes the poison”, which means everything could lead to toxicity
when taken in too high a dose, these findings serve as a rough guideline for the
safe usage of the medicinal plants. The bigger the gap between the dose
required to exert a certain biological activity (effective concentration, EC50) and
IC50 value, the safer it is to be used.
It must however be noted that the data generated from this study is only
preliminary, further tests are required for extensive toxicological evaluation.
Keywords: toxicity, medicinal plants, safety evaluation
246
INTRODUCTION
Malaysia consists of Sabah and Sarawak (the East Malaysian States located at
the northern coast of Borneo) and West Malaysia, also known as Peninsular
Malaysia. Malaysia has a population of 28 million and 3.5 million are
indigenous people, which is about 12% of the total population. In Peninsular
Malaysia, the number of Orang Asli is around 150,000, representing 0.6% of
the national population. Orang Asli is the Malay term used to refer to
indigenous people in Peninsular Malaysia. Majority of the Orang Asli live in
rural and remote areas. They depend substantially on natural resources in the
ecosystem for subsistence, e.g. by hunting, fishing, gathering, swidden farming,
arboriculture, and trading forest products. The forest is the source of their
livelihood; it shapes their customs and culture, defines their identity, and
constitutes their local environment (Kardooni et al. 2014).
The Orang Asli also depends on the forest as a source of health care
remedies. Many plants in the forest contain medicinal values that are able to
cure various health problems. This traditional knowledge on medicinal plants is
inherited from generation to generation. As modernisation takes place, there is
a rapid change in the natural environment. Compounded with economic,
political, and cultural change on a global scale, many indigenous knowledge
systems are in danger of vanishing. Since the traditional knowledge of Orang
Asli might disappear without a trace, there is a need to identify and document
such knowledge as a basis for scientific research and enhance the
understanding. The Malaysian Ministry of Natural Resources and Environment
funded a study, Pangkalan data pengetahuan tradisi Orang Asli di
Semenanjung Malaysia, in an attempt to document the commonly used
traditional knowledge of the Orang Asli, specifically the traditional knowledge
on medicinal plants. Besides documentation, selected plant species were also
evaluated for various therapeutic and cosmeceutical potentials. The study
begins in 2007 and is still on-going up to this date. In this paper, the safety
aspect of 50 selected methanol extracts at cellular level will be discussed.
METHODOLOGY
A total of 15 medicinal plant species that are used by 5 communities of Orang
Asli – Jah Hut, Jakun, Kanaq, Semoq Beri, and Bateq were collected during
2011–2012. Voucher specimen for each species was prepared and stored at the
Traditional Knowledge (TK) Specimen Room, Forest Research Institute Malaysia
(FRIM).
247
The methanol extracts were prepared by separating the plants into
different parts, dried, and ground. Ground samples were then soaked in
methanol for 3 days before evaporation to produce dried extracts. The dried
extracts were stored at -20C until bioassay evaluation.
WRL-68 (liver-like) and Vero (kidney-like) cells were used for in vitro
toxicological evaluation. Cells (1 × 104 cells/well) were exposed to the extract
for 72 hours and viability was measured by the MTT (3-(4, 5-dimethylthiazolyl2)-2, 5-diphenyltetrazolium bromide) assay as described by (Mosmann 1983).
Dose-response curve for each extract was plotted and the median inhibitory
concentration (IC50) was determined by non-linear regression using a variable
slope model (GraphPad Prism 6, GraphPad Software Inc., California, U.S.A).
A total of 50 methanol extracts were prepared from different parts of the 15
plants species. Figure 1 shows the number of extracts categorised according to
their IC50 values. Majority of the extracts exhibited IC50 values ranging between
20-625 g/ml. Only a few extracts were cytotoxic at low concentration (IC50 <
20 g/ml) or did not negatively affect cell viability (IC50 > 625 g/ml).
Upon closer inspection, out of the 50 extracts tested, only 72% exhibited
similar toxicity towards both Vero and WRL-68 cells (Figure 2a). The breakdown
is as follows: 6.0% of the extract had IC50 < 20 g/ml; 30.0% with IC50 20-100
g/ml;
24.0%
with
IC50
100-625
g/ml;
12.0%
with
IC50 > 625 g/ml and 28.0% (14 extracts) had different toxicity level towards
Vero and WRL-68 cells. For this remaining 14 extracts, 21.4% was cytotoxic
towards Vero cells with IC50 value < 20 g/ml; 42.9% with IC50
20-100 g/ml; 28.6% with IC50 100-625 g/ml and 7.1% did not cause more
than 50% of cell death at the concentrations tested (Figure 2b). Meanwhile, for
the toxicity towards WRL-68 cells, 14.3% demonstrated IC50 values
< 20 g/ml; 35.7% with IC50 20-100 g/ml; 42.9% with values between
100-625 g/ml; and 7.1% with values > 625 g/ml (Figure 2b).
248
Number of extracts
25
Vero
WRL-68
20
15
10
5
0
< 20 g/ml
20-100 g/ml
100-625 g/ml
> 625 g/ml
IC50 category
Figure 1. Number of extracts categorised according to their IC50 values.
< 20 g/ml
6%
(a)
> 625
g/ml
12%
20-100
g/ml
30%
100-625
g/ml
24%
Vero 
WRL-68
28%
(b)
> 625 g/ml
7.1%
> 625 g/ml
7.1%
< 20 g/ml
14.3%
< 20 g/ml
21.4%
100-625
g/ml
28.6%
100-625
g/ml
42.9%
20-100
g/ml
42.9%
20-100
g/ml
35.7%
WRL-68
Vero
Figure 2. Percentage of extracts categorised according to their IC50
values, (a) extracts with similar toxicity towards Vero and WRL-68 cells,
(b) extracts with different toxicity levels towards Vero and WRL-68 cells.
Figure 1: Number of extracts categorised according to their IC50 values.
249
In order to determine whether an extract is toxic or otherwise, it is best
to compare the IC50 value determined from a toxicological study to the median
effective concentration (EC50) for a desired therapeutic activity. This is because
toxicity is dose-dependent. Everything could become toxic when the dose
taken is too high. For example, water is an essential element for all living things
but if a person drinks too much water, this could lead to water intoxication.
Hence the saying “the dose makes the poison”. An extract has favourable
safety profile when the difference between IC50 and EC50 values is large.
CONCLUSIONS
Few medicinal plants commonly used by the Orang Asli appear to have very
low IC50 value (< 20 g/ml). These plants may cause toxicity should the dosage
required for desired therapeutic effect exceeds the mentioned IC50 value.
Contrary to popular belief that all natural are safe, the usage of plants with
very low IC50 value must be exercised with caution. The findings obtained from
this study are only preliminary and serve as a rough guideline for the safe
usage of the medicinal plants. Further tests are required for extensive
toxicological evaluation.
REFERENCES
Kardooni, R., Kari F., Yahaya, S.R. & Yusup, S.H. (2014). Traditional Knowledge
of Orang Asli on Forests in Peninsular Malaysia. Indian Journal of Traditional
Knowledge: 13: 282─291.
Mosmann, T. (1983). Rapid colourimetric assay for cellular growth and survival
application to proliferation and cytotoxicity assays. J. Immuno. Methods: 54:
55─63.
250
TOXICITY STUDY
ARBORESCENS)
OF
SARAWAK
WILD
PEPPER
ROOT
(PIPER
HP Chua1, M Syahida2 & D Nicholas1
1
Food Science Technology Research Centre, MARDI Kuching Station, Lot 411,
Block 14, Santubong Road, Petra Jaya, 93050 Kuching, Sarawak
2
Food Science Technology Research Centre, Malaysian Agriculture Research
and Development Institute (MARDI), Selangor
Tel: 082-446 531
Fax: 082-446 529
ABSTRACT
Sarawak wild pepper root (Piper arborescens) has been used in cooking as
health food for many generations and is generally considered as safe, mainly
due to its natural origin but without sufficient scientific studies. This study was
aimed to evaluate the effect of wild pepper root extract on Sprague-Dawley
rats subjected to sub-acute 28-day repeated doses. The rats were given wild
pepper root extract at the dose of 1 (low dose), 2 (medium dose) and 5 (high
dose) g/kg body weight. The control group was given distilled water. Based on
the body weight, organ relative weight and serum biochemistry parameters in
sub-acute toxicity results, wild pepper root showed no noticeable gross toxicity
in all treated Sprague-Dawley rats even though the white blood cell volume,
alanine aminotransferase (ALT) level and creatinine concentration were
significantly lower in rats treated with wild pepper root as compared to the
control group. This indicates that the no-observed-adversed-effect-level
(NOAEL) of wild pepper root extract is 5 g/kg body weight per day.
Keywords: wild pepper root, sub-acute, toxicity, Sprague-Dawley rat
INTRODUCTION
The genus Piper belongs to Piperaceae family, comprises five genera and
approximately 1,400 species, distributed in the tropical and subtropical regions
(dos Santos et al. 2001). Various phytochemical investigations of Piper species
have led to the isolation of various classes of physiologically active compounds
(Parmar et al. 1997).
Biodiversity prospecting of Sarawak’s wild Piper species is of great
importance since many have been long known to possess numerous health
enhancement properties, particularly among the local communities in
Malaysian Borneo (Tawan et al. 2002). Of the many indigenous Piper species
251
known in Sarawak, P. arborescens (lada hutan) is one of the most common
species widely distributed in the lowlands and hill rainforest. The perennial
roots and vines of this wild pepper have a peppery flavour, which is the reason
it is given the general name “Sarawak wild pepper root” or “akar lada liar
Sarawak”. The decoction of wild pepper root is used as household remedy and
reputed to help in treating rheumatism, gout, gastrointestinal diseases,
asthma, bronchitis, improvement of metabolism and provide body warmth
(Chai et al. 1989).
Wild pepper root has been consumed for many generations and is
generally considered as safe, mainly due to its natural origin but without
sufficient scientific studies. Since safety is regarded as one of the important
aspects in herbal product development, the objective of this study is to
evaluate the effect of wild pepper root extract on Sprague-Dawley rats
subjected to in vivo sub-acute (28-day) repeated doses.
Dried wild pepper roots were procured from the local market in Kuching,
Sarawak. The dried roots were cleaned thoroughly with fresh water and dried
at 45C, and then cut into small pieces using a blender. The extract was
prepared using traditional decoction method i.e. by boiling with water at the
ratio of 60 g/1.5 L for 30 minutes. Extracts were then filtered and stored at 4°C
in an airtight bottle until use.
All procedures concerning the use of animals were approved by MARDI
Animal Ethic Committee. All rats were allowed to acclimatize for a week before starting
the study as stated in WHO (1993) guidelines. The sub-acute 28-day repeated dose
study was performed based on Ryu et al. (2004). Five male Sprague-Dawley rats
weighing 200-250 g per group were given distilled water (control dose), 1 g/kg (low
dose), 2 g/kg (medium dose) and 5 g/kg (high dose) body weight of extract via forced
feeding. The low dose weight in this study was formulated based on the normal dose
used in human consumption. On day 29, all rats were fasted overnight for
approximately 12 hours. Blood samples were collected from posterior vena cava under
light ether anaesthesia. Body weight, organ relative weight, blood hematology and
serum biochemistry parameters were analysed.
252
Physical and Body Weight Assessment
Oral administration of wild pepper roots extract did not induce any mortality at
any dose level tested within the 28-day repeated feeding study. All the treated
rats also did not show any sign of gross toxicity, adverse pharmacological
effects or abnormalities with respect to hair coat, eye colour, rashes and skin
irritation. The gross examination of internal organs also did not show any
inflammation, changes in colour and no abnormal spots occurred as compared
to control group.
Serum Biochemistry
1. Liver Function
Administration of wild pepper root extract did not show any significant
difference in all parameters except for serum alanine aminotransferase (ALT)
(Table 1). Significant decreased in serum ALT level was observed in rats fed on
high dose wild pepper root extract – the higher the dose, the lower the ALT
level. However, the differences were not significant among all the three
treated groups. This implies that the extract might not cause any toxic effect on
the liver even at high dose. Reduction of ALT level is important for instilling
protection of the liver from oxidative damages (Jie et al. 2014).
2. Lipid Profile
There was also no significant difference observed in all the plasma lipid profiles
in all the three treated groups as compared to the control (Table 2). Even
though no significant difference was found compared to the control, an
increase in high density lipoprotein (HDL) level in the low- and medium-dose
groups, decrease of cholesterol level in medium- and high-dose group and
decrease of low density lipoprotein (LDL) level in low-, medium- and high-dose
groups were observed. These are indicators that the extract might have the
potential to reduce cardiovascular risk factors.
253
Table 1. Effects of wild pepper root extract intake on liver function in rats
(n = 5) after 28-day repeated dose
Parameter
Control
Low Dose
Medium
High Dose
Dose
ALT (U/l)
58.33 ±
53.83 ±
48.67 ±
45.67 ±
8.12a
12.51ab
3.83ab
6.44b
AST (U/l)
114.76 ±
109.10 ±
104.96 ±
99.43 ±
a
a
a
11.12
22.22
4.13
9.73a
ALP (U/l)
290.17 ±
215.00 ±
220.83 ±
243.83 ±
a
a
a
102.00
63.59
66.09
22.50a
Bilirubin
2.20 ± 1.31a 2.58 ± 0.49a 2.55 ± 0.63a 3.12 ± 0.45a
(µmol/l)
Total Protein
72.97 ±
71.98 ±
71.76 ±
73.23 ±
(g/l)
4.24a
4.52a
3.14a
3.92a
Albumin
45.42 ±
43.82 ±
44.32 ±
45.35 ±
a
a
a
(g/l)
1.55
1.18
1.04
1.87a
Globulin
27.67 ±
28.17 ±
27.17 ±
27.83 ±
(g/l)
3.83a
5.19a
2.71a
2.48a
a
a
a
Albumin/
1.68 ± 0.27
1.61 ± 0.35
1.65 ± 0.17
1.63 ± 0.12a
Globulin
ratio
Glucose
4.80 ± 2.02a 5.50 ± 0.43a 5.75 ± 0.83a 5.53 ± 0.39a
(mmol/l)
*Values are means ± standard deviation (SD) for five rats in each group. Means with
the same letter in the same row are not significantly different (p<0.05).
Table 2. Effects of wild pepper root extract intake on lipid profile in rats
(n = 5) after 28-day repeated dose
Parameter
Control
Low Dose
Medium
High Dose
Dose
Cholesterol
1.53±0.68a
1.55±0.31a
1.23±0.45a
1.21±0.15a
(mmol/l)
HDL
0.44±0.15a
0.51±0.15a
0.45±0.08a
0.38±0.09a
(mmol/l)
LDL (mmol/l) 0.83±0.51a
0.80±0.52a
0.55±0.41a
0.50±0.16a
a
a
a
Trigliseride
0.56±0.15
0.57±0.05
0.54±0.07
0.60±0.17a
(mmol/l)
254
3. Kidney function
There was no significant difference in urea concentrations among all groups of
rats. However, the creatinine concentrations were significantly lower in all rats
treated with wild pepper root as compared to the control group (Table 3). The
significantly lower creatinine levels suggest decreased breakdown of muscle
creatinine. This may be attributed to the metabolic property of wild pepper
root extract that causes increase rate of renal clearance of creatinine, hence
decreasing its serum concentration in the treated rats.
Table 3. Effects of wild pepper root extract intake on serum biochemistry
parameters for kidney function in rats (n = 5) after 28-day repeated dose
Parameter
Control
Low Dose
Medium
High Dose
Dose
Urea
7.28±0.52a
7.49±1.59a
7.23±0.58a
8.30±1.51a
(mmol/l)
Creatinine
58.83±3.31a 53.17±3.19b 53.00±3.10b 50.83±3.43b
(µmol/l)
CONCLUSION
Based on the body and organ relative weight, blood hematology and serum
biochemistry parameters in this sub-acute toxicity study, wild pepper root
showed no noticeable gross toxicity in all treated Sprague-Dawley rats. The noobserved-adversed-effect-level (NOAEL) of wild pepper root extract is
5 g/kg body weight per day. Even though no significant difference was found
between treated and the control group, there were indicators that wild pepper
root extract might have the potential to reduce cardiovascular risk factors.
Further studies are needed to identify the active phytochemicals present in
wild pepper root to elucidate their cardiovascular protection activities.
ACKNOWLEDGEMENTS
The authors thank the Ministry of Higher Education, Malaysia for their financial
support via Fundamental Research Grant Scheme (FRGS), research grant
number 03─03─03─064 EA 001.
255
REFERENCES
Chai, P.P.K., Lee, B.M.H. & Othman, I. (1989). Native Medicinal Plants of
Sarawak. Report No. FB 1. Sarawak: Forest Botany Unit, Forest
Department. Pp. 53–55.
dos Santos, P.R.D., Moreira, D.D., Guimaraes, E.F., & Kaplan, M.A.C. (2001).
Essential Oil Analysis of 10 Piperaceae Species from the Brazilian
Atlantic Forest. Phytochemistry 58: 547–551.
Jie, M., Cheung, W.M., Yu, V., Zhou, Y., Tong, P.H. & Ho, J.W.S. (2014). Anti-Proliferative
Activities of Sinigrin on Carcinogen-Induced Hepatotoxicity in Rats. PLoS ONE
9(10): e110145. doi:10.1371/journal.pone.0110145.
Parmar, V.S., Jain, S.C., Bisht, K.S., Jain, R., Taneja, P., Jha, A., Tyagi, O.D.,
Prasad, A.K., Wengel, J. Olsen, C.E. & Boll, P.M. (1997). Phytochemistry
of the Genus Piper. Phytochemistry 46(4): 597–673.
Ryu, S.D., Park, C.S., Baek, H.M., Baek, S.H., Hwang, S.Y. & Chung, W.G. (2004).
Anti-Diarrheal and Spasmolytic Activities and Acute Toxicity Study of
Soonkijangquebo, A Herbal Anti-Diarrheal Formula. Journal of
Ethnopharmacology 91: 75–80.
Tawan, C.S., Ipor, I.B., Fashihuddin, B.A. and Sani, H. (2002). A Brief Account on
the Wild Piper (Piperaceae) of the Crocker Range, Sabah. ASEAN
Review of Biodiversity and Environmental Conservation (ARBEC).
http://www.arbec.com.my/ pdf/art6julysep02.pdf.
WHO. (1993). Research Guidelines for the Evaluation of the Safety and Efficacy
of Herbal Medicines. Manila: WHO. Pp.35–40.
256
IN SILICO PREDICTION OF DRUG LIKENESS AND ADMET PROPERTIES OF
SOME CENTELLA COMPOUNDS
O Asiah, J Mohd Haffiz, MM Siti Syarifah, MA Nordatiakma, Y Nurhanan, H Lili
Sahira & P Mazura
Forest Research Institute Malaysia (FRIM), 52109 Kepong, Selangor
Tel: 03-6279 7671
Fax: 03-6272 9805
ABSTRACT
Centella asiatica is widely used in the Ayurvedic medicine, traditional African
medicine, traditional Chinese medicine and also becoming popular in the west.
The plants are probably been used since prehistoric times for various medicinal
and cosmetic purposes. The wide spectrum of pharmacological activities could
be associated with its secondary metabolites such as triterpenoid saponins.
The variation in secondary metabolites has been observed in C. asiatica with
identical phenotypes and growth conditions, depending on plant origin. Thus,
one could expect significant differences in active constituents in samples
originating from different countries. The chemical composition is also
important in contributing to the biological activities, hence ascertain the final
product application. C. asiatica extracts have been used for many ailments
which led to successful treatments, where many commercial formulations
available contain madecassoside and asiaticoside in different ratios. In fact,
clinical trials were performed on undefined alcohol or aqueous extracts and
defined extracts comprising certain combinations of triterpenoids. Centella
asiatica is reported to have low toxicity and no known side effects within the
recommended dosage have been reported. Even in oral application,
asiaticoside at 1 g/kg body weight showed no toxicity. Nevertheless, reports
showed C. asiatica may cause local allergic reactions and burning when used
externally and subcutaneously. In this study, molecular properties, drug
likeness and ADMET properties of triterpenoids from C. asiatica namely
asiaticoside, asiatic acid, madecassoside, madecassic acid, brahminoside,
brahmoside and madasiatic acid were evaluated using computational tools.
The results will suggest whether the compounds have potential to be further
developed as a lead compound or therapeutic agent as well as provide
information on its mechanisms of toxic action.
Keywords: Centella asiatica, in silico, prediction, drug likeness, ADMET
257
INTRODUCTION
Centella asiatica or locally known as pegaga is probably been used since
prehistoric times for various medicinal and cosmetic purposes. The plant has
been used to treat many kinds of diseases such as gastrointestinal disease,
gastric ulcer, asthma, wound healing and eczema (Brinkhaus et al. 2000). The
wide spectrum of pharmacological activities could be associated with its
secondary metabolites such as triterpenoid saponins.
In this study, some compounds from C. asiatica (Figure 1) have been
subjected for the prediction of drug likeness and ADMET properties using free
web based tools in order to identify the potential candidate for drug
development. Computational approaches such as ligand-based and structurebased methods have become an efficient approach for drug discovery as well
as providing insight on pharmacokinetic properties (absorption, distribution,
metabolism, excretion and toxicity (ADME/Tox) (Kortagere & Ekins 2010). As
such in silico techniques could speed up research by identifying new molecules
with possible medical applications prior to laborious experiments and
expensive preclinical studies. Asiatic acid, asiaticoside and madecassic acid are
known to be clinically effective on systemic scleroderma, abnormal scar
formation and keloids (Hong et al. 2005). Asiatic acid and asiaticoside were
reported to posses a neuroprotective effects against amyloid beta (Xiong et al
2008; Mook-Jung et al. 1999). A pentacyclic triterpene, asiatic acid has also
been reported to induce apoptosis of various cancer cells (Park et al. 2007).
a
d
b
e
c
f
g
Figure 1. Compounds from C. asiatica comprising a (asiaticoside), b (asiatic
acid), c (madecassoside), d (madecassic acid), e (brahminoside), f (madasiatic
acid) and g (brahmoside).
258
The prediction of drug likeness, physicochemical and ADMET properties were
performed using free online server based on Lipinski’s rule of five (ROF)
including
OSIRIS
property
explorer
(http://www.organicchemistry.org/prog/peo/), Molinspiration (http://www.molinspiration.com),
OpenTox (http://toxtree.sourceforge.net) and OCHEM (Sushko et al. 2011). The
structure of asiatic acid, asiaticoside, madecassic acid and madecassoside were
obtained from PubChem (Bolton et al. 2008), while madasiatic acid,
brahminoside and brahmoside structures were acquired from FooDB
(foodb.ca/compounds/).
Drug Likeness Prediction
The molecular properties prediction of asiatic acid, asiaticoside, madecassic
acid, madecassoside, brahminoside and brahmoside were evaluated using web
based tools such as Molinspiration and OSIRIS property explorer.
Molinspiration checked whether the compound complies to the Lipinski’s ROF,
whereas OSIRIS predicted the drug relevant properties. The results obtained
from both tools were given in Table 1. Most of the compounds complied with
the ROF except for asiaticosside and madecassoside, which violated three of
the criteria i.e. molecular weight, number of hydrogen donor and acceptor.
Madecassic acid, brahminoside and brahmoside did not comply for molecular
weight which was larger than 500 g/mol. While, asiatic acid and madasiatic acid
had no violation towards the Lipinski criteria hence making them the most
potential compounds for drug development. According to Lipinski’s rule, an
orally active drug has no more than one violation of the above criteria. OSIRIS
property explorer predicted compounds madecassic acid, brahminoside and
brahmoside with the highest drugscore of 0.265, followed by asiatic acid
(0.234), madasiatic acid (0.232), madecassoside (0.205) while the lowest
drugscore is asiaticoside with 0.194.
ADMET Prediction
Toxicity prediction using OSIRIS property explorer revealed that all the
compounds were not mutagenic, tumorigenic, irritant and reproductive
effective. Prediction on toxicological hazard using OpenTox revealed that the
compounds shared common properties such as Verhaar scheme, start
biodegradability, skin irritation/corrosive and Kroes TTC which are class 5, class
259
2, not corrosive and class 1, respectively (Table 2). It is also observed that
asiaticoside and madecassoside shared the same profile for Cramers rule and
eye irritation, which are class III and not eye lesion R36 respectively. The other
compounds showed class I and not lesions R34, R35, R36 or R4. In general, all
compounds were predicted as substance that would not be expected to be a
safety concern and non inhibitor for all cytochrome P450 enzymes, except for
asiaticoside that could not be predicted for Cyp2C19.
CONCLUSION
In silico prediction on drug likeness and ADMET properties revealed that
compounds from C. asiatica such as asiatic acid, madasiatic acid, madecassic
acid, brahminoside and brahmoside displayed drug-like and good ADME
properties. Furthermore, they do not show toxicity risk and were categorised
as substances that would not be expected to be a safety concern. Hence, the
compounds may have a potential to be further developed as promising lead
compounds for drug development.
ACKNOWLEDGEMENTS
We would like to thank the Ministry of Agriculture and Agro-Based Industry
Malaysia for the financial support.
REFERENCES
Sushko, I., Novotarskyi, S., Korner, R., Pandey, A.K., Rupp, M., Teetz, W., et al.
(2011). Online Chemical Modeling Environment (OCHEM): Web
Platform for Data Storage, Model Development and Publishing of
Chemical Information. Journal of Computer-Aided Molecular Design:
25; 533─554.
Bolton, E., Wang, Y., Thiessen, P.A., & Bryant, S.H. (2008). PubChem: Integrated
Platform of Small Molecules and Biological Activities. Chapter 12 Pp
217-241 in Wheeler R.A. & Spellmeyer D.C. (Eds.). Annual Reports in
Computational Chemistry Vol 4. Oxford, UK: Elsevier.
Brinkhaus, B., Lindner, M., Schuppan, D. & Hahn, E. G. (2000). Chemical,
Pharmacological and Clinical Profile of the East Asian Medical Plant
Centella asiatica. Phytomedicine: 7; 427─428.
260
foodb.ca/compounds/
Kortagere, S. & Ekins, S. (2010). Troubleshooting Computational Methods in
Drug Discovery. Journal of Pharmacological and Toxicological Methods:
61; 67─75.
Hong, S.S., Kim, J.H. & Shim, C.K. (2005). Advanced Formulation and
Pharmacological Activity of Hydrogel of the Titrated Extract of C.
asiatica. Arch Pharm Res: 28(4); 502─508.
http://www.organic-chemistry.org/prog/peo/
http://www.molinspiration.com
http://toxtree.sourceforge.net
Xiong, Y., Ding, H., Xu, M. & Gao, J. (2009). Protective Effects of Asiatic Acid on
Rotenine- or H2)2-induced Injury in SH-SY5Y cells. Neurochem Res:
34(4); 746-754.
Mook-Jung, I., Shin, J.E., Yun, S.H., Huk, K., Koh, J.Y., Park, H.K., Jew, S.S. & Jung,
M.W. (1999). Protective Effects of Asiaticoside Derivatives Against
Beta-Amyloid Neurotoxicity. Journal of Neuroscience Research: 58(3);
417─425.
Park, B.C., Paek, S.H., Lee, Y.S., Kim, S.J., Lee, E.S., Choi, H.G., Yong, C.S.& Kim,
J.A. (2007). Inhibitory Effects of Asiatic Acid on 7, 12dimethylbenz[a]anthracene and 12-0-tetradecanoylphorbol 13acetate-induced Tumor Promotion in Mice. Biolo Pharm Bull: 30(1);
176─179.
261
Table 1. Drug likeness properties calculated by Molinspiration and OSIRIS property explorer.
Asiaticoside
Molinspiration
miLogP
0.37
TPSA
315.2
No. of atoms
67
Mwt (g/mol)
959.1
No. of hydrogen 19
acceptor
No. of hydrogen 12
donor
No. of violations 3
No. of rotatable 10
bonds
3
Volume (Å )
875.9
OSIRIS Property Explorer
cLogP
-0.423
logS
- 4.75
Mwt (g/mol)
958 .0
Druglikeness
-12.73
Drugscore
0.194
Mutagenic
Tumorigenic
Irritant
Reproductive eff
Asiatic acid
Madecassic
acid
Madecassoside
Brahminoside
Madasiatic
acid
Brahmoside
4.7
97.98
35
488.7
5
3.8
118.2
36
504.7
6
-0.55
355.4
68
975.1
20
3.8
118.2
36
504.7
6
4.96
97.98
35
488.7
5
3.8
118.2
36
504.7
6
4
5
13
5
4
5
0
2
1
2
3
10
1
2
0
1
1
2
487.8
495.8
883.9
495.8
487.6
495.8
4.223
-5.205
488.0
-5.998
0.234
3.371
-4.806
504.0
-5.998
0.265
-1.213
-4.413
974.0
-12.837
0.205
3.371
-4.806
504.0
-5.998
0.265
*Grey colour=violation on Lipinski ROF; green colour= drug-conform behaviour.
262
4.297
-5.313
488.0
-3.893
0.232
3.371
-4.806
504.0
-5.998
0.265
Table 2. Toxicological hazard classification and ADME prediction results by OpenTox and OCHEM
OpenTox
Cramers rule
Verhaar scheme
Start biodegradability
Skin
irritation/corrosion
Eye
irritation/corrosion
Kroes TTC (Threshold
Toxicological Concern)
OCHEM
CYP450
(CYP3A4)
CYP450
(CYP2D6)
CYP450
(CYP2C19)
CYP450
(CYP2C9)
CYP450
(CYP1A2)
modulation
modulation
modulation
modulation
modulation
Asiaticoside
Asiatic acid
Madecassic acid
Madecassoside
Brahminoside
Madasiatic acid
Brahmoside
High (Class III)
Class 5 (Not
possible
to
classify according
to these rules)
Class
2
(persistent
chemical)
Not corrosive
Low (Class I)
Class 5 (Not
possible
to
classify according
to these rules)
Class
2
(persistent
chemical)
Not corrosive
Low (Class I)
Class 5 (Not
possible
to
classify according
to these rules)
Class
2
(persistent
chemical)
Not corrosive
High (Class III)
Class 5 (Not
possible
to
classify according
to these rules)
Class 2 (persistent
chemical)
Not corrosive
Low (Class I)
Class 5 (Not
possible
to
classify according
to these rules)
Class
2
(persistent
chemical)
Not corrosive
Low (Class I)
Class 5 (Not
possible
to
classify according
to these rules)
Class
2
(persistent
chemical)
Not corrosive
Low (Class I)
Class 5 (Not
possible
to
classify according
to these rules)
Class
2
(persistent
chemical)
Not corrosive
Not eye irritation
R36
1
(Substance
would not be
expected to be a
safety concern)
Not lesions R34,
R35, R36 or R41
1
(Substance
would not be
expected to be a
safety concern)
Not lesions R34,
R35, R36 or R41
1
(Substance
would not be
expected to be a
safety concern)
Not eye irritation
R36
1
(Substance
would not be
expected to be a
safety concern)
Not lesions R34,
R35, R36 or R41
1
(Substance
would not be
expected to be a
safety concern)
Not lesions R34,
R35, R36 or R41
1
(Substance
would not be
expected to be a
safety concern)
Not lesions R34,
R35, R36 or R41
1
(Substance
would not be
expected to be a
safety concern)
Noninhibitor
(90%)
Noninhibitor
(93%)
error
Noninhibitor
(94%)
Noninhibitor
(95%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(96%)
Noninhibitor
(94%)
Noninhibitor
(96%)
Noninhibitor
(94%)
Noninhibitor
(93%)
Noninhibitor
(94%)
Noninhibitor
(86%)
Noninhibitor
(93%)
Noninhibitor
(89%)
Noninhibitor
(97%)
Noninhibitor
(96%)
Noninhibitor
(94%)
Noninhibitor
(96%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(95%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(96%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(94%)
Noninhibitor
(95%)
Noninhibitor
(96%)
263
NUTRIGENOMIC EFFECTS OF CURCULIGO LATIFOLIA ON TYPE 2
DIABETIC MODEL
I Nur Akmal1 & I Maznah1,2
1
Nutricosmeceutical and Nutrigenomic Programme, Laboratory of Molecular
Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM
Serdang, Selangor Darul Ehsan, Malaysia
2
Department of Nutrition and Dietetics, Faculty of Medicine and Health
Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan,
Malaysia
Tel: 03-8947 2115
ABSTRACT
The present study evaluated the anti-diabetic effects of Curculigo latifolia
fruit:root. The hypoglycemic and hypolipidemic effects of C. latifolia fruit:root
were investigated in high fat diet (HFD) with combination of low dose
streptozotocin (STZ, 40 mg/kg body weight (b.w)) induced diabetic rats. C.
latifolia fuit:root was orally administered once a day at 50, 100 and 200 mg/kg
b.w for 30 days. Before and after treatments, plasma glucose, lipid and insulin
were measured. The nutrigenomic effects of C. latifolia fruit:root extracts on
genes involved in glucose and lipid metabolisms were further studied on
glucose transporter (GLUT 4), leptin (LEP) and insulin receptor substrate 1
(IRS1) in adipose tissues. Results showed that 200 mg/kg b.w of C. latifolia
fruit:root extracts significantly reduced blood glucose and plasma total
cholesterol (TC), triglyceride (TG) and low density lipoprotein-cholesterol (LDL)
levels. Meanwhile, insulin and high density lipoprotein-cholesterol (HDL) were
increased. In addition, C. latifolia fruit:root extracts also increased the
regulation of GLUT 4 (1 fold), Lep (3 fold) and IRS1 (2 fold) in adipose tissues.
The results suggest that C. latifolia possess significant hypoglycemic and
hypolipidemic effects. Hence, C. latifolia can be used as functional food or
incorporated as a supplement in health-care food for the treatment of type 2
diabetic.
Key words: Curculigo latifolia, gene expression, high fat fed diet, insulin, type 2
diabetes mellitus
INTRODUCTION
Diabetes mellitus (DM) is a metabolic disorder which has become one of the
major worldwide health issues. It has been defined as a continual metabolic
264
disorder which leads to the deficiency in the production of insulin by the
pancreas along with insulin resistance. It has been reported that 371 million
peoples have been diagnosed with diabetes in 2012 and this number is
expected to increase by 2030. This scenario leads to a prediction that by 2030,
diabetes will become the seventh leading cause of death in the world (IDF
Diabetes Atlas Group 2015). In Malaysia, the National Health and Morbidity
Survey (NHMS) 2011 has shown that the prevalence of diabetes has increased
by 31.0% in the space of just 5 years, from 11.6% in 2006 to 15.2% in 2011
(National Diabetes Registry Report 2012).
Several factors have been identified to influence diabetic development such as
hyperglycemia, hyperlipidemia, oxidative stress, excessive advanced glycation
end products (AGEs) and insulin resistant (Yokoyama et al. 2015). Despite of
that, existing drugs for diabetes treatment have also shown some side effects
after prolong intake. Studies have shown that sulfonylureas cause
hypoglycemia, increase in body weight, gastrointestinal disturbance and
headache to the user (Yu et al. 2015). Besides, metformin has been reported to
cause abdominal pain, diarrhea, nausea and lactic acidosis to the diabetic
patient (Wheeler et al. 2013). Other diabetic drug that cause adverse side
effect is thiazolidinediones and according to Oribe et al. (2012) it causes
hepatoxicity after prolongs usage. This has drawn alternative treatment by
using natural products which are more affordable, less toxic and proven to cure
diabetic complications.
Curculigo latifolia is also called lemba or lumbah among local community in
Malaysia. Traditionally, it has been used as sweetener in drinks (Chooi 2006).
This shrub tree consists of berry-like fruit which exhibits both sweet tasting and
taste modifying activities (Kant 2005). The sweet taste of C. latifolia has been
identified and it is due to curculin and neoculin proteins (Ibuka et al. 2006).
Despite C. latifolia is sweet and can be used as alternative sweetener for
diabetic patient, there is less scientific study on C. latifolia as an antidiabetic
agent. This finding could provide an insight on the new properties of C. latifolia
extracts in treating diabetic and its mechanisms. Its also provide an opportunity
to food industries in designing product for diabetic patients.
Preparation of C. latifolia Extract
Preparation of C. latifolia fruit:root extracts were prepared according to
previous method as described elsewhere (Nur Akmal et al. 2013). The
265
concentration of C. latifolia fruit:root extracts used in this study were 50, 100
and 200 mg/kg body weight (b.w).
Preparation of High Fat Diet (HFD)
The normal pellet diet (NPD) from Miba Mansura (Malaysia) are consists of
46% of cornstarch, 26% of palm kernel meal, 4% of soybean oil, 3.5% of
minerals mixture, 1% of vitamins mixture, 0.25% of choline bitartrate and
0.18% of L-cystine. Meanwhile, the high fat diet was formulated based on the
composition provided by Levin et al. (1989). It has been prepared from a
mixture of 50% normal rat chow pellet, 24% of corn oil (Mazola brand), 20% of
full-cream milk powder (Nespray brand from Nestlé) and 6% sugar.
Animal Study
Fifty male Sprague-Dawley rats weighing 160-180 g were purchased from
Perniagaan Usaha Cahaya (Malaysia). Upon received, rats were acclimatized for
a week with free access of water and normal pellet diet. Rats were housed
individually in polypropylene cages and maintained under controlled room
temperature (22±2oC) and humidity (55±5%) with 12:12 h light and dark cycle.
All rats were provided with free access to water and commercially available rat
normal pellet (NPD) prior to acclimatize period. All experimental protocols for
animal care and use were approved by the Animal Care and Use Committee
(ACUC) of Faculty of Medicine and Health Sciences, Universiti Putra Malaysia
(UPM). The diabetic rats were developed with high fat diet and low dose
streptozotocin (STZ, 40mg/kg b.w) for 30 days before being treated with C.
latifolia fruit:root. The rats were randomly divided into seven groups of six
animals each (n=6). Below are the lists of the treatment groups:
Group 1
Group 2
Group 3
Group 4
Group 5
Group 6
Group 7
: Normal (normal pellet diet, non-diabetic, untreated) rats.
: Obese (high fat-fed diet, untreated) rats.
: Diabetic control (high fat-fed diet, induced with STZ, diabetic,
untreated) rats.
: Diabetic test rats (high fat-fed diet, induced with STZ, diabetic
treated with 50 mg/kg b.w of C. latifolia fruit:root extracts.
: Diabetic test rats (high fat-fed diet, induced with STZ, diabetic)
: Diabetic test rats (high fat-fed diet, induced with STZ, diabetic)
: Diabetes test rats (high fat-fed diet, induced with STZ, diabetic)
treated with glibenclamide 10 mg per kg b.w.
266
Treatment was carried for 30 days. At the end of experimental
period, all rats were fasted for 15 hr prior to sacrifice. Blood samples were
collected by cardiac puncture in order to measure glucose, lipid and insulin
levels. Meanwhile, adipose tissues were excised and stored at -80oC prior
used.
Biological Assays
Blood samples were collected using K2EDTA blood collection tube (BD
Diagnostics, New Jersey). Plasma was collected after blood was centrifuged at
3,000 rpm for 10 min. Biological assay such as glucose, total cholesterol (TC),
triglycerides (TG), low density lipoprotein (LDL), high density lipoprotein (HDL)
were measured using Selectra XL clinical chemistry analyzer (Vital Scientific,
Netherlands). Insulin level was measured using rat insulin ELISA kit (Mercodia
AB, Uppsala, Sweeden) with rat insulin as a standard.
Gene Expression Analysis
Total RNA was extracted from adipose tissues using RiboPureTM Isolation of
High Quality Total RNA (Ambion, USA). Frozen tissues were thawed before
starting the disruption procedure. The RNA extraction was carried out as
described in RiboPureTM protocol. The quantification of different express
gene between group treatments was using GenomeLab GeXP Start Kit.
Multiplex primers for GLUT4, Lep and IRS1 were design designed by using
eXpress designer module of the GenomeLab eXpress Profiler software.
Samples were prepared and were added to the appropriate wells of 96-well
sample microplate. All the samples were run triplicates. Besides, all the data
were analyzed using eXpress Analysis software where fragment data is easily
identified. Multiplex genes were normalized with 18S by dividing the peak
area of each gene by peak area of 18S gene. The expression level was
calculated according to this formula:
Fold change = Normalized data of the gene from treated samples
Normalized data of the gene from untreated samples
Statistical Analysis
All data were expressed as mean ± standard deviation and each value
represents a minimum of three (n=3-6) replicate experiments and all assay
conditions were performed in triplicate. Data were analyzed using one-way
analyses of variance (ANOVA) followed by Turkey’s post hoc test. Level of
significance was set at p<0.05.
267
Effect of C. latifolia Fruit:Root on Fasting Plasma Glucose, Lipid and
Insulin Levels
The effect of C. latifolia fruit:root on fasting plasma glucose levels of diabetic
rats are showed in Figure 1. After 30 days of high fat diet with intraperitoneal
injection of STZ (40 mg/kg b.w), fasting plasma glucose levels of diabetic rats in
group 3 were elevated four-fold than normal rats (p<0.05). Treatment with C.
latifolia fruit:root caused significant reduction (p<0.05) of plasma glucose levels
in diabetic rats with 54.8% reduction in group 6 followed by 47.4% in group 5
and 45% in group 4 . Meanwhile, diabetic rats treated with 10 mg per kg b.w
glibenclamide showed 38.5% of reduction of plasma glucose level. These
results compliment with study done by Zheng et al. (2011) and Bansal et al.
(2011) where high fat diet with combination of low dose STZ has closely
mimicked to the natural process of the diabetic and metabolic disturbance in
human diagnose as T2DM. Besides, this combination also causes hyperglycemia
in the model. However, 30 days treatment of C. latifolia fruit:root exhibited
hypoglycemic effect in these diabetic models.
Figure 1. The plasma glucose level
Data are means SD for plasma glucose level at day 0 and day 30 treatment. * are
significantly different at p<0.05 compared with diabetic control i.e. group 3
Indicator: (
) before treatment; ( ) after 30 days of treatment
268
Hyperlipidemia has developed in diabetic rats after 30 days induced with high
fat diet and low dose STZ. Meanwhile, lipid profiles in normal rats were in
normal range. Plasma TC, LDL and TG levels in obese rats (Group 2) were
significantly higher (p<0.05) while HDL level was lower than normal rats (Group
1) after 30 days of study (Table 1). Diabetic control rats in Group 3 also showed
a similar pattern as obese rats where plasma TC, TG and LDL levels increased
56.2%, 52.6% and 75.4% respectively as compared to normal rats. It has been
shown that rats administrated with HFD caused dyslipidemia and other
syndromes in diabetic (Rossi et al. 2010). Besides, defect in insulin secretion
due to STZ also caused defect in lipogenic activity. Insulin plays an important
role in stimulating lipogenesis in mammals, by low secretions of insulin it
implicates of high level of lipid in plasma (Rossi et al. 2010). However, plasma
HDL level in diabetic control rats decreased by 51%. C. latifolia has significantly
reduced (p<0.05) plasma TC, TG and LDL levels. Besides, HDL level also
increased after 30 days treatment in diabetic rats in Group 4, 5 and 6.
Table 1. Lipid profiles of rats group
Group
TC (mmol/l)
TG (mmol/l)
1
1.62 ± 0.18
0.57 ± 0.11
2
3.38 ± 0.16*
1.17 ± 0.36*
3
2.53 ± 0.24
0.87 ± 0.20
4
1.49 ± 0.22*
0.84 ± 0.30
5
1.21 ± 0.16*
0.67 ± 0.13*
6
1.17 ± 0.28*
0.69 ± 0.19*
7
1.35 ± 0.16*
0.59 ± 0.13*
LDL (mmol/l)
0.57 ± 0.42
0.92 ± 0.26
1.00 ± 0.10*
0.85 ± 0.06
0.59 ± 0.38*
0.56 ± 0.13*
0.76 ± 0.24
HDL (mmol/l)
0.53 ± 0.24
0.26± 0.09
0.26 ± 0.15
0.75 ± 0.21*
0.82 ± 0.19*
0.69 ± 0.20*
0.77 ± 0.27*
Data are means ± SD for lipid profiles of normal rats (Group 1), obese rats (Group 2),
diabetic control rats (Group 3), diabetic rats treated with 50 mg/kg b.w of C. latifolia
fruit:root extracts (Group 4), diabetic rats treated with 100 mg/kg b.w of C. latifolia
fruit:root extracts (Group 5), diabetic rats treated with 200 mg/kg b.w of C. latifolia
fruit:root extracts (Group 6) and diabetic rats treated with 10 mg/kg b.w of
Glibenclamide (Group 7) at day 0 and day 30.* are significantly different at p<0.05
compared with diabetic control i.e. group 3.
269
Figure 2. Fasting plasma insulin
Data are means SD for fasting plasma insulin level at day 0 and day 30 treatment. * are
significant different at p<0.05 compared with diabetic control i.e. group 3.
Indicator: (
) before treatment, (
) after 30 days of treatment
This study showed that plasma insulin level decreased in obese rats. Disruption
of pancreatic β-cells by STZ in diabetic rats has caused insufficient insulin
secretion in blood. This STZ is being uptake into pancreatic cells by GLUT2 and
it causes DNA damage via reactive oxygen species generation (Shingo et al.
2012). However, C. latifolia fruit:root extracts in all concentration has
prevented further disruption of cells. There was 16% of insulin increasing in
rats in Group 4 and followed by 13% in Group 5, 12% in Group 6 and 11% in
Group 7. Previous study also indicated that C. latifolia extracts elicited
insulinotropic properties by stimulating insulin secretion either in basal and
glucose.
Effect of C. latifolia Fruit:Root on GLUT4, Lep and IRS1 Genes in
Adipose Tissues
Present results showed that the expression of GLUT4, IRS1 and leptin genes in
obese (Group 2) and diabetic rats (Group 3) were downregulated in adipose
tissues compared to normal group (Group 1) while leptin gene was
upregulated. However, GLUT4 and IRS1 genes were significantly upregulated
higher in treated diabetic rats in Group 6 followed by group 5, 4 and 7. Those
genes are responsible in insulin signalling, glucose transportation and also in
metabolism of glucose and lipid. According to Rakhshandehroo et al.(2010)
high fat diet may down regulates several transcription factors such as nuclear
receptor (PPAR) and sterol regulatory binding proteins (GLUT). This study is
parallel to finding of Rakhshandehroo et al. (2010) because our diabetic model
270
that has been developed through HFD with combination of STZ also shows
similar results. Downregulation of IRS-1 and GLUT4 genes may defect insulin
signaling and glucose uptake activity. Meanwhile, downregulation of leptin
gene may affect glucose and lipid metabolisms in diabetic rats (Wang et al.
2009).
Table 2. Expression of of GLUT4, Lep and IRS1 genes in adipose tissues after 30
days treatment with C. latifolia fruit:root
Group
Group 1
Group 2
Group 3
Group 4
Group 5
Group 6
Group 7
IRS1
1.5
0.5
0.4
1.3
1.8
1.7
1.2
GLUT4
1.4
0.5
0.3
2.1
1.4
1.0
1.3
Leptin
1.0
3.8
0.6
1.6
2.9
3.0
1.1
CONCLUSION
The present study suggests that the anti-diabetic effects of C. latifolia in
diabetic model occur via upregulation of GLUT4, Lep and IRS1 genes in adipose
tissue. Therefore, C. latifolia can be used as functional food or incorporated as
a supplement in health-care food for the preventing and treating of T2DM.
However, further supporting studies to evaluate these promising effects should
be carried out.
ACKNOWLEDGEMENTS
This work was supported by the Fundamental Research Grant Scheme (FRGS)
from the Ministry of Higher Education, Malaysia.
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Bansal, P., Paul, P., Mudgal, J., Nayak, G., Thomas, P.S., Priyadarsini, K.I, et al.
(2011). Antidiabetic, Antihyperlipidemic and Antioxidant Effects of the
Flavonoid Rich Fraction of Pilea microphylla L. in High Fat
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Journal Molecular Biology. 359:148─158.
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Nur Akmal, I., Maznah, I., Muhajir, H., Zalinah, A. and Siti Aisyah, A. G. (2013).
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Complementary
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National
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in STZ-3V Rats is Largely due to Dysfunctional Insulin Signaling through
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Ethnopharmacology 133:531─53.
273
PROTEIN EXPRESSION PATTERN IN SWIETENIA MACROPHYLLA SEED
A Noraliza1, MA Nor Datiakma1, M Norwati1, WK Wan Mohd Aizat2, MN
Normah2, H Nor Asmah.1, N Nadiah Salmi1, NA Nashatul Zaimah1 & Mohd AW
Saifuldullah1
1
2
Institute of System Biology (INBIOSIS), Universiti Kebangsaan Malaysia (UKM),
43600 UKM Bangi, Selangor
ABSTRACT
The application of proteomics on protein expression in plants can be
considered new. A proteome study was performed to analyse the cold-stress
response of Swietenia macrophylla seeds. This present study aimed to evaluate
the overall changes of protein components after being stored in two different
temperatures (10C and 4C) for six months. 2-D gel electrophoresis and spot
identification by MALDI-TOF was performed to monitor changes in the
response to cold-stress conditions. In the experiments, 12 spots with at least
two folds altered expression were found. Some of the proteins identified have
previously been described in cold-stress responses. These differentially
expressed proteins indicates the validity of this proteome approach for better
understanding the function and role of these proteins in cold-stress responses.
In addition, we can also predicts biochemical changes related to medicinal
properties such as anti-inflammatory, anti-mutagenic, antipyretic, antifungal
and anti-tumour activities for this species.
Keywords: Swietenia macrophylla, seeds, proteomics, cold-stress responses
INTRODUCTION
Rapid population growth and progress in industrialization have affected the
natural forests. Huge amount of seeds are needed to overcome the difficulties
in reforestation. One of the problems faced by seed industry is the loss of
quality seed during storage period. Understanding seed storage metabolic
behavior using proteomic approach can greatly ensure a better storage of
tropical seed. Therefore, the present study was aimed at investigating proteins
related to seed deterioration. Proteomic involves large-scale study of proteins
that provides information on complex mixtures of proteins in cells and tissues.
The applications are to analyse functional aspects of proteins such as posttranslation modifications, protein-protein interactions, activities and
structures. Attempts to map and characterize the whole protein networks in a
274
system lead to the recognition of proteome technology as an important
research that complements genome studies.
Plant Materials
Plant species used in this research were collected from Yan, Kedah. Swietenia
macrophylla fruits were stored in room temperature for at least 7 days for the
fruits to crack. The fruits were processed to extract the seeds.
Seed Storage
The seeds were randomly selected and then stored in an airtight aluminum bag
for subsequent studies. The seeds were stored at 10°C and 4°C for a period of 6
months to determine seed germination rate after storage.
Protein Extraction
S. macrophylla embryo from the control and 6-month stored seeds were frozen
in liquid nitrogen and ground into fine powder. Protein was extracted using 9
M urea, 4% CHAPS, 0.5% triton X-100 and 100 mM DTT, and incubated for 2 hr
at
4°C.
The
extract
was
centrifuged
at
12,000 rpm for 10 min. The supernatant was collected to determine the
protein concentration.
Two-Dimensional (2D) Polyacrylamide Gel
Isoelectric focusing (IEF) was performed using immobilized pH-gradient strips
(pH 3-10, pH 4-7 and 7 cm) with PROTEAN IEF CELL (Bio-Rad, USA). A total of
100 g of protein was used for the 7 cm IPG strips in a total volume of 125 L
rehydrating buffer consists of 8 M urea, 2% CHAPS, 15 mM DTT, 200 L
bromophenol blue and 2% ampholytes of pH 3-10. IPG strips were rehydrated
at passive mode for 12 hours at 20°C on PROTEAN IEF System (Bio-Rad, USA).
IEF was performed at 20°C using four different steps (step 1: 200 V for 200 V/h
at slow mode; step 2: 500 V for 500 V/h at slow mode; step 3: 1000 V for 1000
V/h
at
linear
mode;
and
step
4:
4000
V
for
16000 V/h at rapid mode). After IEF, IPG strips were equilibrated using two
steps of equilibration buffer for 15 min each at room temperature. IPG strips
was positioned on top of a vertical 12% polyacrylamide gel and
electrophoresed at a constant current of 52 mA for 15 min, 64 mA for 20 min
275
and finally 72 mA per gel. Gels were stained with silver nitrate and scanned
using ImageScanner (GE Healthcare). The IMAGE MASTER 2D PLATINUM
software by GE Healthcare was used to capture gel images and analysis of all
the 2D gels was performed according to the manufacturer’s instructions.
Vertical and horizontal streakings were also removed and background intensity
was substracted during spot detection. The gels were then matched by
landmarking common spots found in different gels. Normalization was
performed to correct variations in spot size and intensity between gels.
MALDI-TOF/TOF
Protein spots were chosen from the 2-DE gels, excised and identified by mass
spectrometry. Peptides were extracted from the gel plugs by three cycles of
sonication in water, followed by acetonitrile for 15 min. The extracted peptides
were concentrated using vacuum centrifugation and mixed with saturated
matrix. The mixture was spotted onto slides for matrix assisted laser
desorption/ionizing-time of flight (MALDI-TOF) mass spectrometry analysis.
The generated peptide mass values were searched against the Swiss Prot and
NCBInr protein databases.
The present study demonstrated the optimized protein extraction protocol for
S. macrophylla embryo and subsequently the proteomes of two different
sources of seeds i.e. control and stored seeds. This study was an early effort to
establish a platform to identify characteristic proteins involved in the
biochemical and molecular process under cold-stress conditions.
Out of all the proteins detected in the S. macrophylla proteome by
IMAGE MASTER 2D PLATINUM software, nine protein spots were unique to the
control seeds. Two protein spots increased in the stored seeds for both
temperature (10°C and 4°C). Protein spot number four was identified as
hypothetical protein while protein spot number two was identified as
Fosfatase-2a, putative. Protein phosphatase-2A (PP2A) is involved in the
response to temperature changes (Cairns et al. 1994) and hyper osmotic
pressure (Cho et al. 1993). This protein plays an important role in enhancing
the resistance of plants to drought stress. PP2A acts in cell cycle regulation,
signal transduction, differentiation and cell transformation (Sheng et al. 2003).
Hypothetical protein is a protein whose existence has been predicted. To
predict a function for each of their predicted protein coding regions or ORFs,
the amino acid sequence of the ORF is compared against all functionally
276
assigned sequences in protein sequence databases. However, we did not
proceed to predict the function of hypothetical protein detected in the present
study.
Table 1. The differentially expressed protein identified in S. macrophylla under
cold-stress conditions.
Spot
no.
1
2
Protein ID
(accession no)
Chaperon ClpB3,
chloroplastic
(502091111)
Fosfatase-2a, putative
(255572254)
Control
4
Mitogen-activated
kinase 12
(474424521)
Hypothetical protein
(255605376)
6
7
8
9
10
11
12
CHLNCDRAFT_50630
(307109986)
MTR_3g107530
(357466271)
MTR_4g132820
(357480417)
Os06g0622000
(115468994)
PRUPE_ppa000345mg
(462423974)
PRUPE_ppa000345mg
(462423974)
TPA: Hypothetical
protein
ZEAMMB73_456506
(414872482)
TPA: serine/threonine
phosphatase
superfamily
(414873460)
Coverage
(%)
65
Theoretical
molecular
mass/
pI
110647.23/6.69
45
33016.41/
Experimental
molecular
mass/
pI
110749.6/
7.1
33565.4/
+
+
5.38
79
43473.47/
8.45
46
6703.05/
7.3
43788.4/
9
+
6812.5/
+
5
(4°C)
+
+
3
(10°C)
+
+
11.88
42
88895.51/
5.73
8.5
89296.6/
7.5
67
37374.81/
5.20
37465.9/
8
57
19718.55/
5.37
19991/
7.8
58
90106.46/
7.53
90848.1/
4.1
75
142260.60/8.89
143482.1/
7.7
66
142260.60/8.89
143482.1/
7.1
58
15037.51/
11.41
15199.1/
8
54
26297.56/
5.24
26679.9/
7
+
+
+
+
+
+
+
+
+
277
CONCLUSION
The proteins identified in the present study may have roles in the regulation of
cell cycle, metabolic pathway and cell defense mechanism or seed maturation.
Changes in their composition could lead to the possibility of protein
deterioration. Information generated from this study will give an impact on the
future of forestry practices and management of seed storage. Further research
can also be conducted by monitoring the possibility of protein-protein
interaction on secondary metabolites changes after seed storage related to the
medicinal value of this species.
REFERENCES
Sheng, W., Hua, X.Y., Sheng, W.W. & Li, X.B. (2003). Protein Phosphatase 2A: Its
Structure, Function and Activity Regulation. Acta Biochimica et
Biophysica Sinica 35(2): 105–112.
Cairns, B.R., Kim, Y.J., Sayre, M.H., Laurent, B.C. & Kornberg, R.D. (1994). A
Multisubunit Complex Containing the SWI/ADR6, SWI2/SNF2, SWI3,
SNF5, and SNF6 Gene Products Isolated from Yeast. Proc. Natl. Acad.
Sci. USA 91: 1950–1954.
Cho, M.H., Shears, S.B., & Boss, W.F. (1993). Changes in Phosphatidylinositol
Metabolism in Response to Hyperosmotic Stress in Daucus carota L.
Cells Grown in Suspension Culture. Plant Physiology 103(2): 637─647.
278
PROTEOME PROFILES OF SEEDS FROM SWIETENIA MACROPHYLLA
AFTER COLD STORAGE
A Noraliza1, MA Nor Datiakma1, Z Fadzlinah1, M Norwati1, WK Wan Mohd.
Aizat2 & MN Normah2
1
Forest Research Institute of Malaysia (FRIM), Kepong, 52109, Selangor,
Malaysia, 2 INBIOSIS, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor
Tel: 03-62797658 Fax: 03-62804623 E-mail: [email protected]
ABSTRACT
Swietenia macrophylla (mahogany) is currently gaining popularity in Malaysia
for its medicinal value. In neighboring countries, the mahogany fruit is referred
to as the essential ingredients of vitamins and natural remedies. The seeds
have many acclaimed medicinal properties such as anti-inflammatory, antimutagenic, antipyretic, antifungal and anti-tumour activities. The seeds can be
obtained from local markets. The present study was undertaken to provide the
temporal patterns and simultaneous changes in protein accumulation after
certain storage condition to examine the seeds deterioration status. The
proteome profiles of seeds underwent storage of 3 months at 4oC was
observed. Proteome profiling experiment via 2-Dimensional Gel
Electrophoresis (2-DE) had detected at least 240 protein spots expressed in
fresh samples and 12 proteins were differentially expressed after storage. The
results will provide a fundamental reference for further studies on the
regulation of protein expression during storage.
Keywords: proteome, seeds, Swietenia macrophylla, cold storage, 2Dimensional Gel Electrophoresis (2-DE)
INTRODUCTION
Proteomics involves large-scale study of proteins that provides information on
complex mixtures of proteins in cells and tissues. The application among others
is to analyse functional aspects of proteins such as post-translation
modifications, protein-protein interactions, activities and structures. Proteins
are very important parts of living organisms, as they are the main components
of the physiological metabolic pathways of cells (Chevalier 2010). Proteomics is
very useful for studies of storage protein deposition in seeds, evaluation of the
effects of growing conditions, monitoring of desirable or undesirable specific
protein components, and assessment of many other parameters affecting the
overall seed (protein) quality (Miernyk 2014; Miernyka & Hajduchb 2011)
279
With the aim of profiling proteins in seed samples, the focus of our
study is to develop a method for the 2-D PAGE analysis of Swietenia
macrophylla seed protein which allows the visualization of a maximum of
proteins from the sample. Preparation of high-quality protein from plant has
been problematic using standard protocols due to high salts, polysaccharides,
lipids and proteases. The improved techniques in protein extraction and 2-D
PAGE enable comprehensive protein visualization on 2-D PAGE gels.
Protein sample preparation
The seed embryos were excised and the protein was homogenised in lysis
buffer containing 9 M urea, 100 mM DTT, 4% CHAPS dan 0.5% triton X-100.
Protein concentration was determined using the 2-D Quant Kit (Amersham
Biosciences, Uppsala, Sweden). The quantification protocol was performed as
described by the manufacturer using Bovine Serum Albumin (BSA) as protein
standard. A standard calibration curve of absorbance values over several
protein concentrations was constructed.
Two Dimensional - Polyacrylamide Gel Electrophoresis (2D PAGE)
Protein sample (50 μg) was diluted in rehydrating buffer containing urea (6 M),
SDS (2%), Tris/HCL pH 8.8 (0.375 M), gliserol (25%) dan DTT (2%) and applied to
7 cm IPG strips of pH 3–10 overnight. The strips were subjected to two-step
equilibration in buffers containing urea (6 M), SDS (2%), Tris/HCL pH 8.8 (0.375
M), gliserol (25%) and DTT (2%) and followed by 250 mM iodoacetamide and
applied to the 12 % SDS-PAGE gels. The second dimension electrophoresis was
performed by SDS-PAGE with an SE 600 Ryby (GE Healthcare, Sweden)
electrophoresis unit. Gels were stained with silver stain according to
Berkelman and Stenstedt (1998). Digital images of the analytical gels were
acquired and analysed quantitatively for differentially expressed proteins using
ImageMaster 2D Platinium 7.0 analysis software (GE Healthcare, Sweden).
From the protein extraction, seeds that underwent 3 months storage at 4oC
yielded a total protein of 5.2 ± 0.14 μg/μL. The proteins profiles obtained from
2-Dimensional Gel Electrophoresis (2-DE) were established using immobilized
pH-gradient (IPG) strips pH 3–10, 7 cm.
280
Gel image analysis was performed by comparing the occurrence of
every spot among the sets of protein profiles from fresh sample and cold
storage treatment. Proteome profiling experiment via 2-DE detected at least
240 protein spots expressed in fresh samples and 12 proteins were
differentially expressed after storage. From the analysis of 12 differentially
expressed protein spots (parameters: >1.5 folding value & p<0.05); seven
proteins were down-regulated and five proteins were up-regulated. Figure 1
shows the representative gel images of fresh (b) and seeds underwent cold
storage. While Figure 2 shows the representatives’ gel spots of selected protein
of interest.
Bil
Bil
Bil
Embrio
Embrio
biji
biji
benih
benih
kawalan
kawalan
11
+
+
22
Embrio
Embrio
biji
biji
benih
benih
yang
yang
disimpa
disimpa
n
n (4°C)
(4°C)
3
3 bulan
bulan
1
1
11
2
2
+
+
22
Bil
Embrio
biji
Bil
Embrio
+
+
benih
biji
33
kawalan
benih
Embrio
3
3
biji
Embrio
benih
biji
43
yang
benih
34
Figure 1. Representative
image of protein profiles
kawalan disimpa
yang
+ 4°C. disimpa
months storage +
at
n (4°C)
44
n (4°C)
4
3
bulan
(a)
45
5
3 bulan
11
+
+
55
22
3
66
3
++
+
+
+
Embrio
biji
Bil
Embrio
Bil
Bil Embrio
Embrio
benih
biji
biji
biji
kawalan
benih
benih
benih
kawalan
kawalan
kawalan
+
+
+
+
65
+
+
++
+
+
7
6
6
++
+
of mahogany seeds after 3
+++
+
+
+
+
+
++
++
(b)
+
+
+
+
+++
5
+
+
Embrio
biji
Embrio
Embrio
Embrio
benih
biji
biji
biji
yang
benih
benih
benih
disimpa
yang
yang
nyang
(4°C)
disimpa
disimpa
n3
(4°C)
bulan
disimpa
bulan
n3
(4°C)
n
(4°C)
33
bulan
bulan
+
+
+
+
+
+
+
+
+ of interest.
+ (a) down+
+
Figure 2. Representatives
gel spots of selected
protein
+
8
8
+
7
7
regulated
(b) up-regulated protein spots.
4 and +
+
4
88
5
5
99
+
+
+
+
+
+
+
+
+
+
+
281
7
7
9
9
8
810
10
+
+
+
+
+
+
+
+
CONCLUSION
Further work on identification of the 12 differentially expressed proteins will
be performed. The protein identification data will be useful for fundamental
reference to further studies on the regulation of protein expression during
storage.
ACKNOWLEDGEMENTS
We wish to thank FRIM Seeds Technology Laboratory (MTBB), Forest
Biotechnology Division and Biomolecule Research Laboratory, Natural Products
Division personnel for their technical assistance in this study.
REFERENCES
Berkelman, T., & Stenstedt, T., (1998). 2-D Electrophoresis- Principle and
Methods. Sweden: Amersham Bioscience.
Chevalier, F. (2010). Highlights on the Capacities of "Gel-based" Proteomics.
Proteome Science. 8:23
Miernyk A.J., (2014). Seed
Biology. 1072:361─77
proteomics.
Methods
of
Molecular
Miernyka A. J. & Hajduchb M., (2011). Seed proteomics. Journal of Proteomics.
74. 389 – 400.
282
EFFECT OF pH ON ADSORPTION OF ORGANIC ACID AND PHENOLIC
COMPOUNDS IN NONI (MORINDA CITRIFOLIA L.)
H Haslaniza, WA Wan Yaakub & MY Maskat
School of Chemical Sciences and Food Technology, Faculty of Science &
Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor
ABSTRACT
Adsorption of model solution containing organic acids (octanoic and hexanoic
acid) and phenolic compounds (rutin, scopoletin and quercetin) on a weak base
anion exchange Amberlite IRA 67 resin was studied in a model system. This
research was carried out to understand the effect of pH on adsorption of multicompounds studied for its further use in actual system (fruit system). It was
shown that the pH dependence of adsorption capacity of Amberlite IRA 67
resin had similar increasing trend on organic acids (hexanoic and octanoic acid)
and phenolic compounds (rutin, scopoletin and quercetin) studied as pH values
increased. Similar trend was also found in multi-compounds solution of
phenolic compounds in total phenolic content (TPC) and antioxidant activity
(FRAP and DPPH). The findings obtained in this study will help to gain better
understanding of the complex mechanisms of ion exchange resin and
adsorption process involving multi-compounds in fruit (noni) system.
Keywords: adsorption, ion exchange resin, organic acids, phenolic compounds
INTRODUCTION
Morinda citrifolia L. or noni has been used traditionally to treat a broad range
of diseases for over 2000 years (Dixon et al. 1999). About 160 phytochemical
compounds have already been identified in noni plant, with the major
micronutrients being phenolic compounds, organic acids and alkaloids (Wang &
Su 2001). The unpleasant odour of noni extract is contributed by medium chain
fatty acids such as decanoic, hexanoic and octanoic acids (Norma et al. 2004).
Recently, more attention has been focused on the role of natural antioxidants,
in particular, phenolic compounds. Scopoletin is a characteristic phytochemical
in noni, while rutin and quercetin are bioactive flavonoids (Deng et al. 2010).
Due to the beneficial role of antioxidants, it is important that deacidification
did not reduce the antioxidant activity.
283
This study will focus on the adsorption behaviour of organic acids
(octanoic acid and hexanoic acid) and phenolic compounds (rutin, scopoletin
and quercetin) onto Amberlite IRA 67 resin in a model system. Apart from that,
this research is also important to obtain more information on the interaction of
organic acids constituents and phenolic compounds with the resin using multicompounds system.
Materials
Weak base anion exchanger Amberlite IRA 67 (food grade) and standard
compound of octanoic acid, hexanoic acid, rutin, scopoletin and quercetin were
purchased from Sigma Aldrich Corporation. Methanol (purity > 99.9%), HPLC
grade was purchased from Fisher Scientific.
Adsorption Studies Of Organic Acids And Phenolic Compounds
The adsorption studies were carried out in Erlenmeyer flasks where 0.1 g of the
resin and 0.02 l of each samples which were octanoic acid (250 mg/l), hexanoic
acid (100 mg/l), rutin (25 mg/l), scopoletin (25 mg/l), and quercetin (25 mg/l),
were added without pH adjustment. The flasks were agitated in the orbital
shaker at 120 rpm for 180 min to achieve equilibrium. After which the
adsorption of each of the organic acids was measured using a Gas
Chromatography. While adsorption of each of the phenolic compounds was
measured using HPLC. The amount of organic acids and phenolic compounds
adsorbed at equilibrium, qe(mg/g), were calculated based on the equation
below:
Where Co and Ce are the concentrations of the each organic acids and phenolic
compounds at the beginning and in the equilibrium, respectively (mg/L); V is
the volume of the solution (L); w is the mass of the dry anion exchanger (g).
pH Determination
pH determination was carried out on all samples using a pH meter (Model PB10, Sartorius Basic Meter, Germany).
284
Determination of Ferric Reducing Assay (FRAP)
FRAP assay was conducted according to the methods of Benzie and Strain
(1996). The results were expressed as μmol/ g fresh weight (FW) sample.
Determination of Free Radical Scavenging using DPPH Method
The antioxidant activity of all juices were evaluated based on free radical
scavenging effect according to the methods of Akowuah et al. (2005).
Percentage of free radical scavenging activity was calculated based on the
formula below:
% inhibition of DPPH = [Abs control – Abs sample/ Abs control] x 100
Determination of Total Phenolic Content
Total phenolic content of noni juice was determined using Folin-Ciocalteau
reagents as described by Singleton and Rossi (1965). The results were
expressed as mg GAE/g FW.
Effect of Multi-compounds of Organic Acids
In this model system, multi-compounds solution was considered as actual noni
juice with identical values of pH. As can be seen in Figure 1, there was no
significant difference detected in both control samples A and B for hexanoic
and octanoic acid. If compared to the adsorption capacity obtained in multicompounds solution, the trend observed for both compounds were similar. It
shows an increase in adsorption capacity as the pH values increased. The
highest adsorption capacity for hexanoic and octanoic acid were significantly (p
< 0.05) obtained at pH 10 to 11 and pH 9 to 11, respectively.
The adsorption capacity increases with increasing pH values. It can be
explained by the reduction of the positive repulsive interactions between
functional groups at high pH. Therefore, the adsorption of low acidic solution is
more favoured. The same phenomenon happened in the study done by Berbar
et al. (2012) in the adsorption of polyethyleneimine using anion exchange
resin.
285
Figure 1. The effect of organic acids of multi-compounds (A0 = 25 ppm; OA =
250 ppm; HA = 100 ppm) using Amberlite IRA 67 resin at different pH (Resin
amount 100 mg; agitation rate: 120 rpm)
Effect of Multi-compounds of Phenolic Compounds
Figure 2 presents selected phenolic compounds (scopoletin, rutin and
quercetin) in multi-compounds system at different pH range between 2 to 11
including control samples. From the observation, there was no significant
difference between both control samples (A and B) in scopoletin and quercetin
concentration but significantly (p<0.05) decreased in treated samples with pH
modification. On the other hand, there was no significant difference found in
rutin concentration in control samples (A and B) and also rutin solution at pH of
2 to 4. As the pH increased, each phenolic compound seems to decrease until it
reached pH 11. The lowest concentration of each phenolic compound was
determined at pH 11 which were significantly difference (p<0.05) among other
treated samples. The same observations were made in antioxidant acitivity
(DPPH and FRAP) and TPC (data not shown) where the activity decreased as the
pH value increased.
One reason for the loss of antioxidant activity observed at higher pH
value could be due to higher activity of polyphenoloxidase resulting in higher
phenol degradation. Phenolic compounds are known as powerful chain
breaking antioxidants (Shahidi & Wanasundara 1992). The phenolic compounds
may contribute directly to antioxidative action (Duh et al. 1999).
286
Figure 2. HPLC determination of multi-compounds (A0 = 25 ppm; OA = 250
ppm; HA = 100 ppm) using Amberlite IRA 67 resin at different pH (Resin
amount 100 mg; agitation rate: 120 rpm)
CONCLUSION
It was shown that the pH dependence of adsorption capacity of weak base
anion exchange Amberlite IRA 67 resin had similar increasing trend on organic
acids (hexanoic and octanoic acid) and phenolic compounds (rutin, scopoletin
and quercetin) studied as pH values increased. Similar trend was also found in
multi-compounds solution of phenolic compounds in total phenolic content
(TPC) and antioxidant activity (FRAP and DPPH). The findings from the present
study are important for further investigation to be applied in actual system.
ACKNOWLEDGEMENTS
The authors would like to thank The Ministry of Education (MOE) for the
scholarship, The Ministry of Science, Technology and Innovation (MOSTI) for
financing the project under the ERGS/1/2013/STWN03/UKM/02/1 grant and
Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia.
REFERENCES
Akowuah, G. A., Ismail, Z., Norhayati, I. & Sadikun, A. (2005). The Effects of
Different Extraction Solvents of Varying Polarities on Polyphenols of
Orthosiphon stamineus and Evaluation of The Free Radical-Scavenging
Activity. Food Chemistry 93(2): 311–317.
287
Benzie, I.F.F. & Strain, J. J. (1996). Ferric Reducing Ability of Plasma (FRAP) as a
Measure of Antioxidant Power: The FRAP Assay. Analytical
Biochemistry 239: 70-76.
Berbar, Y., Amara, M. & Kerdjoudj, H. (2012). Effect of Adsorption of
Polyethyleneimine on the Behaviour of Anion Exchange Resin. Procedia
Engineering 33: 126–133.
Deng, S., West, B.J. & Jensen, C.J., 2010. A Quantitative Comparison of
Phytochemical Components in Global Noni Fruits and Their Commercial
Products. Food Chemistry, 122(1), 267–270.
Duh, P. D., Tu, Y. Y., & Yen, G. C. (1999). Antioxidant Activity of Water Extract of
Harn
Jyur
(Chyrsanthemum
morifolium
Ramat).
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und-Technologie-Food
Science
and
Technology 32: 269–277.
Dixon, A. R., McMillen, H. & Etkin, N. L. (1999). Ferment This: The
Transformation of Noni, A Traditional Polynesian Medicine (Morinda
citrifolia, Rubiaceae). Economic Botany 53: 51–68.
Norma, H., Normah, A., Ahmad, A.W., Rohani, M.Y., Muhammad Gawas, M. &
Sharizan, A. (2004). Reducing The Smelly Compounds (Caproic, Caprylic
and Capric acids) in Noni by Treating The Juice with Activated Charcoal
Powder. Proceeding of the National Food Technology Seminar.
Shahidi, F., & Wanasundara, P. K. J. P. D. (1992). Phenolic Antioxidants. Critical
Reviews in Food Science and Nutrition 32: 67–103.
Singleton, V. L. & Rossi, J. A. (1965). Colorimetry of Total Phenolics with
Phosphomolybdic-phosphotungstic Acid Reagents. American Journal of
Enology and Viticultur 16: 144─158.
Wang, M.Y. & Su, C. 2001. Cancer Preventive Effect on Morinda citrifolia
(Noni). Annals of the New York Academy of Sciences 952:161─168.
288
EFFECT OF ORGANIC-BASED FERTILIZER RATE AND PLANTING DISTANCE
ON BIOMASS YIELD OF BELALAI GAJAH (CLINACANTHUS NUTANS)
AG Rosnani1 & M Syahida2
1
Horticulture Research Centre, 2Food Technology Science Research Centre,
MARDI Headquarters, P.O. Box 12301, 50774 Kuala Lumpur, Malaysia
Tel: 03-89536191 Fax: 03-89536195 E-mail : [email protected]
ABSTRACT
Clinachanthus nutans is one of the known cultivated herbs in Malaysia.
Locally known as belalai gajah or Sabah snake grass, this small shrub belongs
to the family of Acanthacea and native to topical Asia. In Thailand, this plant
has long been traditionally used to treat skin rashes, herpes simplex virus and
varicella-zooster lesions. Recently, it has gained popularity among Malaysian
especially for treating cancers. The was considered the useful plant part of
belalai gajah (BG) due to the phytochemicals and flavonoids content. Thus, a
study was conducted to determine optimum fertilizer rate and planting
distance for increasing biomass leaf yield of BG. The study was carried out in
MARDI Serdang, Selangor. Four different rates (0, 100, 200 and 300 kg/ha) of
commercial organic-based fertilizer (NPK 10:5:5) were used in the experiment.
The plants were planted at different planting distances which were 40 cm x 30
cm, 40 cm x 40 cm, 40 cm x 50 cm, and 40 cm x 60 cm. The plants were
harvested every two months as a ratoon crop in three times of harvesting
process. Results showed the interaction between fertilizer rate and planting
distance at first harvest with the optimum fertilizer rate at 220 kg/ha and
planting distance at 40 cm x 40 cm. It produced, fresh yield and dry yield of
300 and 80 kg/ha respectively. The second harvest showed no interaction
between fertilizer rate and planting distance on biomass yield. The optimum
fertilizer rate was at 200 kg/ha which obtained 473 kg/ha of fresh yield and
118 kg/ha of dry yield. Meanwhile, biomass yield at planting distance between
40 cm x 30 cm and 40 cm x 40 cm was not significantly different but better
than two other distances. At the third harvest, the highest biomass yield (888
kg/ha of fresh yield and 224 kg/ha of dry yield) was at fertilizer rate of 300
kg/ha and similar trend was observed at second harvest on planting distance.
It can be concluded that fertilizer rate (NPK 10:5:5) for BG is 210 ± 10 kg/ha for
first and second harvest and 300 kg/ha for third harvest, and BG can be
cultivated using planting distance of 40 cm between row and 35 ± 5 cm within
row.
Keywords: organic-based fertilizer, planting distance, biomass yield,
Clinacanthus nutans
289
INTRODUCTION
Belalai gajah (Clinacanthus nutans is one of the highly valued herbs that has
been identified for developement under Entry Point Project (EPP1). Belalai
gajah (BG) can be traditionally used for diabetes mellitus, fever, diarrhoea and
dysuria (Uawonggul et al. 2011). In recent years, various pharmacological
properties such as immunomodulatory (Sriwanthana et al. 1996), cytotoxic
(Thongrakard & Tencommao 2010), anti-oxidant (Yong et al. 2013) and antiviral (Sakdarat et al. 2009) have been previously reported. BG is a small shrub
that native to tropical Asia countries and a member of the Acanthaceae family.
The plant grows well under the scattered forests and bushes on the
low altitude area up to 620 meters. In recent years, the plant has gained
popularity among Malaysian especially for treating cancers. The leaf is the
most useful plant part due to the various phytochemicals and flavanoids
content. Thus, a study was conducted to determine optimum fertilizer rate
and planting distance to increase the biomass leaf yield of BG.
The field experiment was conducted in MARDI Serdang, Selangor. The plants
were propagated using stem cuttings. Stem was cut into 2 – 3 nodes and was
placed in germination trays (104 plugs) containing mixture of subsoil and
Holland peat at the ratio of 3:1. The germination tray were exposed under 25
% shade for better germination rate and the seedlings were ready to be
transplanted after 30 days. Land preparation for field planting was done by
one round of disc-ploughing about 25 – 30 cm deep and after 1-2 week, land
was rotovated for another round. The soil beds were raised about 30 cm
(height), 1.2 m (width) and 4.5 m (length). The beds were later covered by
plastic mulching (UV silver shine) to control weeds. Planting holes were set up
a day before transplanting and one seedling was transplanted into each hole.
The seedlings were planted at a spacing of 40 cm between rows and different
interplant spacing at 30, 40, 50 and 60 cm.
Four different rates of commercial organic-based fertilizer (NPK
10:5:5) were tested, comprising 0, 100, 200 and 300 kg/ha. Drip irrigation
system was used to supply water to the seedlings at two times daily. Pest and
disease were controlled using commercial pesticide and fungicide. Meanwhile,
weeds in each planting holes were controlled manually. Harvesting was done
as ratoon crop in every two months for three times per cycle. The plants were
harvested about 15 cm from soil surface and the yield was separated
according to plant parts such as leaf and stem. They were dried immediately
290
to avoid further deterioration.
Parameters of growth and biomass yield (fresh and dry) were
recorded and statistically analyzed using SAS software (SAS 9.1, 2002). The
significant means were separated by Duncan’s multiple range test (DMRT) at
P≤0.01. The experiment was designed in a split plot with fertilizer rate as main
plot and planting distance as sub-plot. The treatments were conducted in four
replications.
The interaction between fertilizer rate and planting distance was observed only
on biomass yield at first harvest but no interaction for the second, third and
total harvest. Fresh yield of BG at first harvest had increased with the
increasing rate of fertilizer for 40 cm x 40 cm and 40 cm x 50 cm. After the
application rate of 220 kg fertilizer/ha at 40 cm x 40 cm and after 200 kg
fertilizer/ha for 40 cm x 50 cm, the fresh yield had decreased. However, the
highest yield was at 40 cm x 30 cm and 40 cm x 40 cm planting distance. The
interaction showed that the optimum fertilizer rate was 220 kg/ha with
planting distance at 40 cm x 40 cm. At these rates and distances, fresh yield
and dry yield were at 300 kg/ha (Figure 1) and 80 kg/ha (Figure 2) respectively.
2
2
2
*S1, y=219.26 + 0.4776x (R = 0.95), S2, y=186.07 + 1.992x – 0.0062x (R =0.83),
2
2
2
S3, y=106.92 + 0.407x – 0.0009x (R =0.95), S4, y=90.19 + 0.3009x (R =0.96).
Figure 1. Interaction between fertilizer rate and plant spacing on fresh yield of
belalai gajah at first harvest
291
2
2
2
*S1, y=58.84 + 0.1123x (R = 0.93), S2, y=45.24 + 0.517x – 0.0016x (R =0.85),
2
2
2
S3, y=27.48 + 0.1117x – 0.0003x (R =0.87), S4, y=22.93 + 0.0745x (R =0.96).
Figure 2. Interaction between fertilizer rate and plant spacing on dry yield of
belalai gajah at first harvest
Results at second harvest showed that no interaction between
fertilizer rate and planting distance on biomass yield. The results only focused
on main effect of fertilizer rate and planting distance. The highest fresh yield
(473 kg/ha) and dry yield (118 kg/ha) of BG at second harvest was under
fertilizer of 200 kg/ha and 300 kg/ha (Table 1). Meanwhile, biomass yield
(Table 2) at planting distance between 40 cm x 30 cm and 40 cm x 40 cm
showed no significant difference but better than two other distances. The
fresh yield for both distances were 472 kg/ha and 449 kg/ha, and dry yield
were 117.27 kg/ha and 113.2 kg/ha respectively.
At the third harvest, the highest biomass yield (888 kg/ha of fresh
yield and 224 kg/ha of dry yield) was at fertilizer rate of 300 kg/ha. Similar
trend was observed with second harvest on planting distance of 40 cm x 30 cm
and 40 cm x 40 cm (Table 2).Total yield from all harvests showed that biomass
yield was highest at fertilizer of 300 kg/ha for both fresh and dry yield. The
results also showed that planting distance of 40 cm x 30 cm and 40 cm x 40 cm
was better than 40 cm x 50 cm and 40 cm x 60 cm.
292
Based on research done by Mengel (1982), the highest yield increment
occurred at the lower rate of nitrogen fertilizer applied but with successive
applications of nitrogen, the yield increments become progressively smaller.
On the other hand, according to the Acquaah (2001), when the nutrient are
present in non-limiting amounts, plants have a tendency to use more than they
need. Thus, up to a certain point, uptake of nutrients does not translate into
increased biomass yield or plant productivity. While a research done by
Maheshwar (2005) on stevia in Karnataka, India showed that application of 105
kg N ha-1, 30 kg P ha-1 and 45 kg K ha-1 recorded significantly higher dry leaf
yield. It was due to maximum number of leaves per plant and branches per
plant as compared to lower doses of N under loamy soil. The interaction
between fertilizer rate and planting distance occurred in this study is probably
due to the difference of fertilizer amount received by the plants and the
number of plant population at different plant spacing. There were increments
on both fresh and dry yields with later harvests probably due to increase in
plant maturity and increasing in number of new branches and leaves after
several ratoon were done.
Table 1. Fresh and dry yield of BG leaves at different fertilizer rate for harvest
2, 3 and total harvest
Fertilizer
Rate
(kg/ha)
Dry Yield (kg/ha)
Fresh Yield (kg/ha)
Harvest 2
Harvest 3
Total
Yield
Harvest 2
Harvest 3
Total
Yield
0
301c
442d
899d
76c
111d
558d
100
377b
662c
1233c
94b
167c
806c
200
473a
754b
1481b
118a
189b
937b
300
527a
888a
1637a
131a
224a
1075a
Means with the same letter for each harvest are not significantly different by DMRT at p< 0.01.
Table 2. Fresh and dry yield of BG leaves at different plant spacing for every
harvest and total harvest
Plant
Spacing
Fresh Yield (kg/ha)
Dry Yield (kg/ha)
Harvest 2
Harvest 3
Total
Yield
Harvest 2
Harvest 3
Total
Yield
40cm x30cm
472a
777a
1540a
117.27a
197.32a
1540.11a
40cm x 40cm
449ab
712ab
1427a
113.2a
178.39ab
1426.91a
40cm x 50cm
399bc
640bc
1175b
99.29b
159.92bc
1175.03b
40cm x 60cm
358c
617c
1110b
89.37b
155.4c
1110.01b
Means with the same letter for each harvest are not significantly different by DMRT at p< 0.01.
293
CONCLUSION
The study concluded that optimum fertilizer rate (NPK 10:5:5) for BG is 210 ±
10 kg/ha for first and second harvest and 300 kg/ha for third harvest based on
biomass yield. It also showed that BG was best cultivated using planting
distance of 40 cm between row and 35 ± 5 cm within row.
ACKNOWLEDGEMENTS
The authors wish to thank Mr. Zakry Al-Asyraf Abd Latiff, Norsyamiza Mat
Yusof and Noor Kamilah Omar for their assistance in implementing the
experiment. This project was funded by MARDI under MEGA Project (P161).
REFERENCES
Acquaah, G. (2001). Principles of Crop Production: Theory, Techniques and
Technology. Pearson Education Inc. p: 246–271.
Maheshwar, H.M. (2005). Effect of Different Levels of Nitrogen and Dates of
Planting on Growth and Yield of Stevia (Stevia Rebaudiana Bert.). M.Sc.
Thesis, Department of horticulture, University of Agricultural Sciences,
Dharwad, Karnatka, India p: 100.
Mengel, K & Kirkby, E.A. (1982). Principles of Plant Nutrition. International
Potash Institute (3th Edition) p: 358─65.
Sakdarat, S., Shuyprom, A., Pietong, C., Ekalaksananan, T. & Thongchai,
S.(2009). Bioactive Constituents from The Leaves of Clinacanthus
Nutans Lindau. Bioorganic and Medicinal Chemistry. 17(5):1857─1860.
Sriwanthana, B., Chavalittumrong, P. & Chompuk, L. (1996). Effect of
Clinacanthus Nutans on Human Cell-Mediated Immune Response InVitro. Thai Journal of Pharmaceutical Sciences. 20(4):261─267.
Thongrakard, V., & Tencommao, T. (2010). Modulatory Effects of Thai
Medicinal Plant Extract on Proinflammatory Cytokines-Induced
Apoptosis in Human Keratinocyte Hacat Cells. African Journal of
Biotechnology. 9(31):4999─5003.
Uawonggul, N., Thammasirirak, S., Chaveerach, A., Chuachan, C., Daduang, J &
Daduang, S.(2011). Plant Extract Activities Against The Fibroblast Cell
294
Lysis by Honey Bee Venom. Journal of Medicinal Plant Research
5(10):1978─1986.
Yong, Y. K., Tan, J.J., Teh, S. S., Mah, S. H., Ee, G. C., Chiong, H. S., & Ahmad,
Z.(2013). Clinacanthus nutans Extracts are Antioxidant with
Antiproliferative Effect on Cultured Human Cancer Cell Lines. EvidenceBased Complementary and Alternative Medicine. p:462─751.
295
THE EFFECT OF PACKAGING TECHNIQUE ON QUALITY OF DRIED MAS
COTEK (FICUS DELTOIDEA)
MS Rosalizan1, D Thiyagu2 & D Mohd. Nazrul Hisham3
1
Horticulture Research Centre, MARDI, Serdang, Malaysia
2
Rice & Paddy Research Centre, MARDI, Serdang, Malaysia
3
Food Technology Research Centre, MARDI, Serdang, Malaysia
Tel: 03-8943 7139 Fax: 03-8942 5786
ABSTRACT
Packaging technique is very important in handling of dried herbs because most
herbs are marketed in dried form. Storage life of dried herbs can be prolonged
by selecting suitable packaging materials while maintaining its quality and
phytochemical content of herbal plant. The objectives of this study are to
identify suitable packaging material for dried mas cotek that could prolong its
storage life and to determine its quality and phytochemical content during
storage. The results showed that moisture content of mas cotek increased
from 45 days storage period onwards for Nylon-PE and LDPE packaging
material, meanwhile the moisture content of mas cotek packed in aluminium
packaging stabilized up to 180 days of storage period. It was observed that
antioxidant activity remained unchanged throughout the storage period for all
types of packaging materials. Total phenols in mas cotek increased from 45 to
90 days of storage period for all types of packaging materials but the
concentration decreased during 135 and 180 days of storage period. The
increased in total flavonoids content can be seen in all packaging materials up
to 90 days of storage period. However, the concentration decreased when
stored up to 180 days in all packaging materials. Based on this study, it is
recommended to use LDPE bag for bulk packaging because it is cheaper,
reliable and readily available. The cost of LDPE bag is comparable with Nylon PE
bag but LDPE bag is much stronger with high tensile strength. Nylon PE is not
suitable for bulk packaging because the material is easily broken and could not
withstand heavy loads. Although aluminium bag is expensive, it provides good
moisture barrier and suitable for retail packaging.
Keywords: mas cotek, packaging, quality
INTRODUCTION
Malaysian rainforest has been reported to host more than 2,000 plants with
medicinal value, and in most village around the country, the older generation
still use myriad brews, poultices and pastes derived from herbal plants for
everything from physical ailments to beauty. The use of herbal plants and their
296
related products have become increasingly important worldwide over the past
two decades. Global trade of natural plant products is projected to triple by
2020, with the herbal medicine market expected to grow by 10 and 20 percent.
On the domestic front, herbal industry in Malaysia is estimated to grow at the
rate of 15% per annum, with the market value rising from 7 billion ringgit in
2010 to 29 billion ringgit in 2020 (Nor Mohamad 2011).
Mas cotek is one of the priority herbs under EPP1 of NKEA in
agriculture cluster. This herb can be planted in most part of Malaysia and
grows well in bris soil. The information on planting technique of this plant has
already been established by MARDI.
Export of the produce would inevitably be in dried form, but for
formulation of product, it can be from fresh or dried raw material. The
parameters for maintaining quality in agricultural produce after harvest are
generally well understood with proper temperature maintenance being of
paramount importance. Recently, much attention has been focused on the
phytochemical content within plant material that may help prevent the onset
of certain cancers and cardiovascular diseases. Herbs are one of them. At this
early stage of our knowledge on the bioefficacy of phytochemical in fruits and
vegetables, it appears the most important are: polyphenolic compounds,
including anthocyanins and flavonols, terpenoids, including carotenoids, and
organo-sulfur compounds such as glucosinolates and cysteine sulfoxides
(Jones et al. 2006). Relatively little is known of the effects of commonly used
postharvest handling procedures on phytochemical content. In most cases,
conditions that contribute to loss of cellular integrity cause a decline in
phytochemical content, but there are important exceptions where processing
and/or cooking sometimes increased both phytochemical content and
bioavailability (Jones et al. 2006). It is well documented that certain
postharvest operation such as drying and storage will affect the phytochemical
content. High temperature of more than 50°C will cause reduction in
phytochemicals (Muller & Heindl 2006). Hence, low temperature drying
between 30°C and 50°C are recommended to protect sensitive active
ingredients (Muller & Heindl 2006). Low temperature storage is very
important to prolong shelf life of horticultural crops such as fruits, vegetables
and herbs. Studies by Lee and Howard (1990) have shown that flavonoids
content in banana declined 40 to 45% during 4 months of storage period.
Storage of green bell peppers for 7 and 14 days at 7°C and 21°C retained 94%
and 78% of carotene, respectively (Matthews et al. 1975). Mas cotek is
perishable and easily deteriorates after few days of harvest, so proper
postharvest handling is needed to retain its phytochemical and prolong its shelf
life. However, the information on the effects of postharvest handling on
phytochemical content of mas cotek is unavailable.
297
In the case of prolonging the storage life of the fresh produce, suitable
packaging materials and temperature need to be established. Shelf life of fresh
produce can be further extended under low temperature storage with proper
packaging materials such as perforated LDPE bag. Mariana et al (2011)
reported that proper packaging material of thyme and rosemary had shown
negligible quality loss up to one year of storage. Similar results also found in
coneflower where freeze-dried sample packed in polyethylene bag, aluminium
foil and nylon polyethylene bag have shown highest retention of bioactive
compound (Lin et al. 2011) Thus, there is a need to evaluate the stability of the
phytochemicals and shelf life under storage conditions in order to meet fresh
and dried market demand for leafy herbs.
Drying and Packaging
The plant (mas cotek) was harvested at Farmer’s farm, Pedas, at optimum
maturity of 12 months after planting. The herb was washed and dried in solar
dryer until the moisture content dropped to 10%. Then, the plant was
transported to Post Harvest Laboratory at MARDI, Serdang and packed in
various types of packaging materials, namely low density polyethylene bag,
Nylon-PE bag and laminated aluminium bag. The thickness of the bag is 0.08
mm. The packed herb was stored for 6 months under ambient condition (room
temperature of 25°C). Each packaging materials contain 100 g (0.1 kg) of dried
leaves. The physical and chemical analyses were done at intervals of 45, 90,
135 and 180 days of storage.
Moisture Content and Phytochemical Analyses
Moisture content was analysed using moisture analyser model AND 300. Total
phenolic content and total flavonoids content were determined by method
described by Marinova et al. (2005). The total antioxidant activity was
determined by the method described by Chan et al. (2008). All analyses were
done in 4 replicates.
Statistical Analyses
Data were analysed by Statistical Analysis System (SAS) software version 8.
Significance differences between means (p<0.05) was determined using LSD
(SAS Inst. 1990).
298
Packaging technique is very important in handling of dried herbs because most
herbs are marketed in dried form. Storage life of dried herbs can be prolonged
by selecting suitable packaging materials while maintaining its quality and
phytochemical content of herbal plant. The results in Figure 1 show moisture
content of mas cotek increased from 45 days storage period onwards for
Nylon-PE and LDPE packaging materials. This result indicates that there was
moisture movement from the surrounding area into the packaged content. It
can be said that, nylon-PE and LDPE were highly permeable to water. However,
moisture content of mas cotek packed in aluminium packaging was stabilized
for all storage periods. So, aluminium can be recommended as a packaging
material for dried mas cotek for longer shelf life. It was observed that
antioxidant activity was stabilized throughout the storage period for all types
of packaging materials (Table 1). Results in Table 2 show the total phenols in
mas cotek was found to increase from 45 to 90 days of storage period for all
types of packaging materials. It might probably due to synthesis of phenols
compound during the storage period. However, the concentration decreased
during 135 and 180 days of storage period. This finding is similar to Lachman et
al. (2003) report in which total polyphenols increased during storage of red and
yellow varieties of onions at 22°C. However, total flavonoid remained
unchanged for all type of packaging materials throughout the storage period
(Table 3). The results of statistical analysis of moisture content, antioxidant
activity, total phenolic and flavonoids contents of mas cotek in different
packaging techniques are presented in Table 4.
Moisture content (% w.b)
12
10
8
6
4
Nylon PE
2
LDPE
Aluminium
0
0
45
90
135
180
Days of storage
Figure 1. The effect of packaging materials on retention of moisture content in
mas cotek
299
Table 1. The effect of different packaging technique on antioxidant activity of
mas cotek
Storage period
0 Days
45 Days
90 Days 135 Days
180
Days
Packaging
technique
Nylon PE
86.61
87.59
84.13
86.36
86.5
LDPE
86.61
87.27
82.54
86.00
86.23
Aluminium
86.61
88.32
82.71
88.11
87.06
Table 2. The effect of different packaging technique on total phenols content
(g/g) of mas cotek
Storage period
0 Days
45 Days
90 Days 135 Days 180 Days
Packaging
technique
Nylon PE
LDPE
Aluminium
377.78
377.78
377.78
447.78
386.85
387.22
508.7
496.48
540.00
329.26
392.78
316.85
371.48
396.00
362.78
Table 3. The effect of different packaging technique on total flavonoids content
(mg/100g) of mas cotek
Storage period
0 Days
45 Days
90 Days 135 Days 180 Days
Packaging
technique
Nylon PE
LDPE
Aluminium
377.78
377.78
377.78
447.78
386.85
387.22
300
508.7
496.48
540.00
329.26
392.78
316.85
371.48
396.00
362.78
Table 4. Statistical analysis of mas cotek in different packaging technique
Response
Total
Total
Total
Moisture
flavonoids
phenolics
antioxidant
content (%
(mg/100g CE) (ug/g GAE)
activity (%)
w.b)
Storage time
n.s
**
**
**
(days)
0
222.57 a
377.78 c
86.59 b
7.58 d
45
206.70 b
407.28 b
87.73 a
8.23 c
90
235.76 a
515.06 a
83.14 c
8.56 b
145
215.05 b
346.30 d
86.85 b
9.72 a
180
217.01 b
376.73 d
86.60 b
8.78 b
Packaging
n.s
n.s
n.s
**
material
Nylon PE
226.92 a
409.96 a
85.74 b
9.34 a
LDPE
209.18 b
407.00 a
86.24 ab
9.12 b
Laminated
222.16 a
396.93 a
86.56 a
6.96 c
aluminium
Storage time
n.s
*
n.s
**
X packaging
material
CONCLUSION
Improper packaging technique could promote bacteria and fungi growth during
storage. So, selecting the best packaging material is crucial to ensure the safety
of herbal product. Based on this study, it is recommended to use LDPE bag for
bulk packaging because it is cheap, reliable and readily available. The cost of
LDPE bag is comparable with Nylon PE bag but LDPE bag is much stronger with
high tensile strength. Nylon PE is not suitable for bulk packaging because the
material is easily broken and could not withstand heavy load. The cost for
aluminium bag is more expensive compared to others but this material provides
good moisture barrier and it is suitable for retail packaging.
REFERENCES
Chan, E.W.C., Lim, Y.Y., Wong, L.F., Lianto, F.S., Wong, S.K., Lim, K.K., Joe, C.E. &
Lim, T.Y. (2008). Antioxidant and Tyrosinase Inhibition Properties of
Leaves and Rhizomes of Ginger Species. Food chemistry 109: 477–483.
301
Lin, S., Sung, J.M., & Chen, C. (2011). Effect of Drying and Storage Conditions on
Caffeic Acid Derivatives and Total Phenolics of Echinacea Purpurea
Grown in Taiwan. Food Chemistry 125: 226–231.
Jones, R., Premier, R. & Tomkins, B. (2006). The Effect of Postharvest Handling
Conditions on Phytochemicals Important for Human Health Contained
in Fruits and Vegetables. In Advances in postharvest technologies for
horticultural Crops. CABI publication. Pp. 1–19.
Lachman, J., Proněk, D., Hejtmánková, A., Dudjak, J. & Pivec, K. (2003). Total
Polyphenol and Main Flavonoid Antioxidants in Different Onion (Allium
cepa L.) Varieties. Hort. Sci. (Prague) 30(4): 142–147.
Lee, Y. & Howard, L.R. (1999). Firmness and Phytochemical Losses in
Pasteurized Yellow Banana Peppers (Capsicum annum) as Affected by
Calcium Chloride and Storage. Journal of Agricuktural and Food
Chemistry 47: 700–703.
Marianna, U., Mauro, M., Marzia, F., Alessandra, D.C. & Roberta, D. (2011).
Influence of Different Stabilizing Operations and Storage Time on the
Composition of Essential Oil of Thyme (Thymus officinalis L.) and
rosemary (Rosmarinus officinalis L.). LWT – Food Science and
Technology 44: 244–249.
Marinova, D., Ribarova, F. & Atanassova, M. (2005). Total Phenolics and Total
Flavonoids in Bulgarian Fruits and Vegetables. Journal of the University
of Chemical Technology and Metallurgy 40(3): 255–260.
Matthews, R.F., Locasio, S.J. & Ozaki, H.Y. (1975). Ascorbic Acid and Carotene
Contents of Pepper. Proc. Florida State of Horticultural Science Society
88: 263–265.
Muller, J. & Heindl, A. (2006). Drying of Medicinal Plants. In Medicinal and
aromatic plants. Springer publication. Pp. 237–252.
Nor Mohamed, Y. (2011). Investment and Business Opportunities in Herbal
Industry. Keynote paper presented at Symposium of Investment and
Business Opportunities in Herbal Industry, 14 April 2011, Renaissance
Hotel, Kuala Lumpur.
302
POTENTIAL OF MEDICINAL PLANTS USED BY THE JAKUN PEOPLE AS
ANTITUBERCULOSIS AGENTS
S Siti Fatimah, M Maryati & AB Mohd Fadzelly
Centre of Research for Sustainable Uses of Natural Resources, Faculty of
Science, Technology & Human Development, Universiti Tun Hussein Onn
Malaysia (UTHM), 86400 Parit Raja, Batu Pahat, Johor, Malaysia
Tel: 07-456 4473
Fax: 07-453 6051
ABSTRACT
Tuberculosis (TB) is a global health problem that causes high rate of mortality
by a single pathogen called Mycobacterium tuberculosis. The effective yet
problematic antituberculosis drugs and the emerging resistant strains of TB
persist to bring challenges in TB medication. Therefore, the investigation of
traditionally used plants by the Jakun people to treat TB and its related
symptoms could be an ideal approach in searching for novel antituberculosis
agents to improve the circumstances. This study was carried out to screen
antimycobacterial activities of eight selected medicinal plants, namely
pengesep (Rourea mimosoides), keruing air (Dipterocarpus sublamellatus),
perut keletung (Thottea praetermissa), hempedu beruang (Thottea
grandiflora), akar ipoh (Stryhnos ignatii), akar empelas (Tetracera macrophylla)
and segindu (Rennellia elliptica), extracted in different solvents (aqueous, 80%
MeOH, hexane, ethyl acetate and 100% MeOH). Antimycobacterial activities
were assessed through zone of inhibition (ZOI) by agar disk diffusion, minimum
inhibitory concentration (MIC) by microbtroth assay and minimum
mycobactericidal concentration (MMC) by conventional plating. In this study, a
model organism of TB strain, Mycobacterium smegmatis mc2 155 (ATCC
700084), was shown to be susceptible against the plant crude extracts by
showing different ranges of antimycobacterial activities. Ethyl acetate root
extract of T. grandiflora exhibited the largest ZOI value of 15.08 ± 0.86 mm
while both hexane and ethyl acetate extracts of D. sublamellatus showed the
lowest MIC and MMC values of 0.78 mg/ml and 3.13 mg/ml respectively. The
results suggest that majority of traditionally used plants by the Jakun people
possess antimycobacterial properties and their active crude extracts have a
great potential in the development of future antituberculosis agents.
Keywords: traditional knowledge, Jakun, medicinal plants, tuberculosis,
antimycobacterial activity
303
INTRODUCTION
The genus Mycobacterium is the typical source of human diseases including
tuberculosis (TB). TB is easily transmitted by air and ingestion of air droplets
containing the viable pathogens into the respiratory system can cause
significant mortality in a population (Tripathi et al. 2005). An ideal anti-TB
regimen is not yet available to combat the resistant strains of TB and the
recommended treatment regimens are problematic (Zumla et al. 2014). For
that reason, the search for at least one potentially new drug derived from
nature should be initiated as proposed by The Stop TB Strategy (WHO 2006).
Traditional knowledge (TK) possessed by different ethnic groups in
Malaysia is important in keeping the multi-ethnic identity of the country (Nijar
2013). An indigenous community, called the Jakun, from Kampung Peta,
Mersing, Johor, is among the diverse groups who are still strongly practicing
their TK with regard to medicinal plants in treatment of various ailments (Chee
2005). Nevertheless, their reliance on TK in general health care and medicine is
declining, particularly among the younger generations. The improving socioeconomic status and the convenience access to modern medicine are seen as
probable factors in the declining use of TK (Seow et al. 2013). This knowledge is
now at the threshold of being endangered and eventually feared to lead to
extinction (Haron & Hamiz 2014). For that reason, Cox (2000) suggested that
scientific validation of traditional claims should continuously be explored to
conserve TK.
Guided by the TK of indigenous people, particularly the Jakun of
Kampung Peta, more potentials in terms of medicinal plants could be
discovered. In this study, eight selected medicinal plants used by the Jakun
people in Kampung Peta were screened for their antimycobacterial activities
against Mycobacterium smegmatis mc2 155 (ATCC 700084), which is a model
organism of TB strain.
Collection and Identification of Medicinal Plants
Prior Informed Consent (PIC) was obtained from the Jakun’s traditional herbal
practitioners (THM) to document their knowledge about medicinal plants used
to treat TB and its related symptoms. Plant samples were collected under a
permit granted by Perbadanan Taman Negara Johor (PTNJ). The samples were
collected from areas mentioned by the THM around Kampung Peta and Taman
304
Negara Johor Endau Rompin in March, 2014. Voucher specimens were
prepared and identification of plant species were verified by Mr. Kamarudin
Salleh (Forest Research Institute Malaysia, FRIM). The species of plants and
parts used were detailed in Table 1.
Extraction of Crude Extracts
All samples were allowed to air-dry (AD) for four weeks and pulverized into fine
powder. The powdered samples were extracted in five solvents by:
1) decoction in distilled water for 15 min; 2) maceration in 80% MeoH for 2
hours with constant agitation and repeated three times; and
3) consecutive maceration in hexane, ethyl acetate and 100% MeOH for 24
hours without agitation and repeated three times. Crude extracts were filtered
through Whatman no. 1 filter paper. Aqueous extracts were lyophilized using
freeze-drier while organic extracts were concentrated using rotary evaporator.
Agar Disk Diffusion Assay
Turbidity of overnight culture was adjusted to 0.5 McFarland standard (1.5×108
CFU/ml). The adjusted inoculum was inoculated onto agar plates containing 25
ml Middlebrook 7H10 agar supplemented with oleic acid, albumin, dextrose,
catalase
(OADC).
Whatman
No.
5
filter
paper
disks
(6 mm diameter) were loaded with 20 μl of crude extracts (100 mg/ml). All
impregnated disks were allowed to dry completely before placing them
equidistantly from each other onto agar plates within 15 min of inoculation of
bacterial culture. Prior to incubation, the plates were kept in biosafety cabinet
to facilitate diffusion of the extracts into the agar for
30 min. The plates were then incubated at 37°C for 3 days. The diameter of ZOI
was measured. Each test was performed twice in triplicates and expressed as
mean ± standard deviation.
Microbroth Dilution Assay
Serial two-fold dilutions of each extract in 50 l of Middlebrook 7H9 broth
medium supplemented with acid, dextrose, catalase (ADC) and 20% Tween 80
were prepared directly in 96-well plates at concentrations of 25 to
0.02 mg/ml for crude extracts. Growth controls containing no antibiotic and
sterility controls without inoculation were also included. The inoculum was
adjusted to 0.5 McFarland standard and diluted to 1:100 to get
5 x 105 CFU/ml, then 50 l was added to the well. The plates were covered with
lids and incubated at 37°C in the normal atmosphere. After 3 days of
incubation, 30 l of 0.01% of resazurin solution was added to each well,
305
incubated overnight at 37°C and assessed for color development. A change
from blue to pink indicates reduction of resazurin due to bacterial growth. The
MIC was defined as the lowest concentration of a sample that prevented this
colour
change.
Prior
to
determination
of
MIC
method,
100 l of broth suspension from the microplate wells that showed no colour
change were inoculated onto fresh agar plates. The agar plates were incubated
for 3 days at 37oC. The MMC was defined as the lowest concentration that did
not harbour any bacterial growth on agar plates. Tests were performed twice
in triplicates.
The results suggest that majority of traditionally used plants by the Jakun
people possess antimycobacterial properties against M. smegmatis and their
active crude extracts have great potential in the development of future
antituberculosis agents. As shown in Table 1, ethyl acetate extract of T.
grandiflora exhibited the largest ZOI value of 15.08 ± 0.86 mm, followed by
ethyl acetate extract of T. praetermissa (12.08 ± 2.15 mm) and aqueous extract
of T. grandiflora (11.33 ± 0.52 mm) while all crude extracts of R. elliptica
showed no ZOI value at all.
Table 1. ZOI when M. smegmatis was treated with plant extracts in different
solvents at 100 mg/ml
Samples/ Part
used
ZOI (mm)
Aq
r
R. mimosoides
s
H. macrocarpa
r
R. elliptica
sb
D. sublamellatus
r
T. praetermissa
r
T. grandiflora
s
S. ignatii
s
T. macrophylla
Aq- Aqueous, Hexstem, sb- stem bark
80%
MeoH
Hex
EA
100%
MeOH
9.58±0.49
8.92±0.20 7.75±0.69
8.08±0.74
7.00±0.32
NI
NI
7.08±0.38
7.75±0.88
6.92±0.38
NI
NI
NI
NI
NI
9.67±0.41
8.00±0.63 9.17±0.26
8.75±0.94
7.58±0.92
8.83±0.26
NI
7.42±0.49
12.08±2.15
8.42±0.92
11.33±0.52
NI
7.25±0.27
15.08±0.86
9.75±0.69
7.42±1.02
NI
7.92±0.86
8.58±0.49
NI
8.00±0.32
8.00±1.00 9.42±1.20
8.42±0.38
7.58±0.38
Hexane, EA-Ethyl acetate, MeOH- Methanol, NI- No inhibition, r- root, s-
As shown in Table 2, aqueous extract of T. macrophylla and both
hexane extracts of D. sublamellatus and T. macrophylla exhibited the lowest
MIC value (0.78 mg/ml), followed by ethyl acetate extract of T. macrophylla
(1.56 mg/ml). 80% MeOH extract of T. grandiflora, ethyl acetate extract of T.
306
macrophylla, and both hexane extracts of D. sublamellatus and T. macrophylla
exhibited the lowest MMC value (3.13 mg/ml).
Table 2. MIC and MMC when M. smegmatis was treated with plant extracts in
different solvents
Samples/ Part used
r
R. mimosoides
s
H. macrocarpa
r
R. elliptica
sb
D. sublamellatus
r
T. praetermissa
r
T. grandiflora
s
S. ignatii
s
T. macrophylla
NA- No activity
MIC/MMC (mg/ml)
Aq
80% MeoH
Hex
EA
3.13/25.0
NA/NA
NA/NA
6.25/NA
NA/NA
NA/NA
NA/NA
0.78/NA
12.5/25.0
6.25/25.0
25.0/NA
0.78/3.13
6.25/12.5
6.25/12.5
3.13/6.25
0.78/3.13
6.25/6.25
6.25/12.5
25.0/NA
3.13/6.25
6.25/6.25
3.13/12.5
3.13/6.25
1.56/3.13
25.0/NA
25.0/NA
25.0/NA
12.5/12.5
6.25/6.25
3.13/3.13
25.0/25.0
12.50/12.5
100%
MeOH
12.5/25.0
25.0/25.0
6.25/6.25
6.25/25.0
6.25/25.0
12.5/12.5
25.0/25.0
12.5/25.0
Aqueous solvent was used to correlate with the traditional mode of
application while the four organic solvents were used to extract a wide range
of plant compounds. The most common solvent used by THMs among the
Jakun people is water (aqueous), which is limited by its inability to extract nonpolar compounds (Masoko & Nxumalo 2013). Although aqueous extracts of
some species such as R. mimosoides, D. sublamellatus, and T. macrophylla
displayed promising inhibitory effect against M. smegmatis, this also explains
why the majority of aqueous extracts in this study were less effective in killing
the strain. Hexane and ethyl acetate were the best extractants resulting in
lower concentration for testing M. smegmatis susceptibility.
CONCLUSION
In summary, a total of 40 crude extracts from medicinal plants used by the
Jakun people in Kampung Peta, Johor has shown interesting antimycobacterial
activities in vitro. Their traditional claims have been scientifically validated to
act as possible antituberculosis agents. Potential species such as T. grandiflora,
D. sublamellatus, and T. macrophylla could be further studied to determine
their bioactive compounds.
ACKNOWLEDGEMENTS
The authors acknowledge Malaysia Ministry of Higher Education (MOHE) for
funding this project through Fundamental Research Grant Scheme (FRGS) Vot
1435, Jabatan Kemajuan Orang Asli (JAKOA), Mr. Kamarudin Salleh from Forest
307
Research Institute Malaysia (FRIM), Perbadanan Taman Negara Johor (PTNJ),
Perbadanan Bioteknologi dan Biodiversiti Negeri Johor (J-Biotech), and the
Jakun people who unreservedly shared their precious knowledge.
REFERENCES
Chee, B.J. (2005). Medicinal Properties and Common Usages of Some Palm
Species in the Kampung Peta Community of Endau-Rompin National
Park, Johor. J. Trop. Med. Plant 6(1): 79–83.
Cox, P.A. (2000). Will Tribal Knowledge Survive the Millennium? Science
287(5450): 44–45. doi:10.1126/science.287.5450.44.
Haron, H. & Hamiz, M. (2014). An Ontological Framework to Preserve Malay
Indigenous Health Knowledge. Advanced Science Letters 20(1): 226–
230. doi:10.1166/asl.2014.5261.
Masoko, P. & Nxumalo, K.M. (2013). Validation of Antimycobacterial Plants
Used by Traditional Healers in Three Districts of the Limpopo Province
(South Africa).
Nijar, G.S. (2013). Traditional Knowledge Systems, International Law and
National Challenges: Marginalization or Emancipation? European
Journal
of
International
Law
24(4):
1205–1221.
doi:10.1093/ejil/cht077.
Seow, T.W., Mohamed, M., Nur, M., Bin, S., Asli, O. & Ehwal, J.H. (2013).
Pembangunan Sosioekonomi Komuniti Orang Asli di Malaysia.
Persidangan Kebangsaan Geografi & Alam Sekitar Kali Ke-4 Anjuran
Jabatan Geografi dan Alam Sekitar, Fakulti Sains Kemanusiaan, hlm.
755–761. Universiti Pendidikan Sultan Idris.
Tripathi, R.P., Tewari,N., Dwivedi, N. & Tiwari, V.K. (2005). Fighting
Tuberculosis: An Old Disease With New Challenges. Medicinal research
reviews 25(1): 93–131. doi:10.1002/med.20017.
WHO. (2006). The Stop TB Strategy. WHO/HTM/TB/2006.368. Geneva.
Zumla, A.I., Gillespie, S.H., Hoelscher, M., Philips, P.P.J., Cole, S.T., Abubakar, I.,
McHugh, T.D., Schito, M., Maeurer, M. & Nunn, A.J. (2014). New
Antituberculosis Drugs, Regimens, and Adjunct Therapies: Needs,
Advances, and Future Prospects. The Lancet. Infectious diseases 14(4):
327–40. doi:10.1016/S1473-3099(13)70328─1.
308
MEDICINAL PLANTS USED FOR WOMEN’S HEALTHCARE AMONG THE
JAKUN COMMUNITY IN KG. PETA: A PRELIMINARY STUDY
I Nur Amalina, M Maryati & AB Mohd. Fadzelly
Centre of Research for Sustainable Uses and Natural Resources, Faculty of
Science, Technology and Human Development, Universiti Tun Hussein Onn
Malaysia, Johor
Tel: 07-4564 473
ABSTRACT
Women in Jakun community have been using medicinal plants to cure their
health problems. It is interesting to note their knowledge in this field. This
preliminary study aims to study the medicinal plants used for Jakun women
healthcare. This research was conducted from January 2015 to June 2015 in
Kg. Peta, Endau Rompin Forest. Four key informants from Jakun community
were chosen to participate in this study. Snowball sampling method was used
to determine the highly experienced practitioner. Semi-structured
questionnaire and informal discussion were conducted. The total number of
medicinal plants that were used for Jakun women’s healthcare in Kg. Peta was
12 species from nine families. Seven species of plants were used by women is
for post-partum recovery. Future investigation and proper documentation
need to be done in order to preserve their knowledge from erosion.
Keywords : Jakun, women healthcare, post-partum recovery
INTRODUCTION
Documentation of medicinal plants for ethnic minority is demanding. Their
valuable knowledge on ethhnobotany might be lost forever if it is unrecorded.
Jakun, or also called Orang Hulu is the dominant population of indigenous
people in Johor. This community also could be found in Pahang as they
originally sited at Endau-Valley (Taylor & Wong 1987). The use of medicinal
plants is still widespread in this community. The community was highly
dependent on medicinal plants in their daily life. The medicinal plants used
traditionally by them had been successfully cure ailments and able to maintain
health. Early documentation work of medicinal plants in Kg. Peta recorded 52
plants species were used to treat various ailments (Taylor & Wong 1987). From
the early record, there is about 12 species of plants that were used to treat
women disorder such as for health tonic, post-natal care, abortifacient,
fertility, and menstrual disorder. Most of research had look into the knowledge
309
of male traditional healers and scholars, without taking into account the
knowledge that is adopted by women (Pfeiffer and Butz 2005; de Boer &
Cotingting 2014). As women are also the main user of the herbs, it is
interesting to note their knowledge on this field. This preliminary study aims to
study the medicinal plants used for Jakun women healthcare.
METHODOLOGY
This preliminary study was conducted from January 2015 to June 2015 in Kg.
Peta, Endau Rompin Forest. Four key informants from Jakun community were
chosen to participate in this study. Snowball sampling method was used to
determine the highly experienced practitioner (d’Avigador et al. 2014). Two
criteria that were taking into account when choosing informants are i) the
recognition that they are local practitioner by the Jakun community, and ii) the
ability to identify plants and explain their uses. Semi-structured questionnaire
was used during the interview session followed by informal discussion during
fieldwork. The available plant specimens were photographed, collected and
processed according to the plant taxonomic method.
From the preliminary study the total numbers of medicinal plants that were
used for Jakun women’s healthcare in Kg. Peta are 12 species from 9 families
(Table 1). Seven species of plants were used by women is for post-partum
recovery. From the result, Rourea mimosoides and Cnestis palala had the same
local name (i.e. pengesep) among Jakun people. These two species belongs to
the same family which is Connaraceae. Most of the mode of administration of
herbal medicine is taken orally in the form of decoction.
The plant part that was mostly used is root. Based on the interview, the
root is the main plant parts used in the Jakun traditional medicine practice.
Among Jakun women, kacip fatimah (Labisia pumila) was used for post-partum
treatment. It could induce uterus shrinking. This is in agreement with the data
collected by Taylor and Wong (1987). The mixture of medicinal plants was also
used for the post-partum treatment. According to the informant, after
childbirth, Jakun women usually drink the mixture of L. pumila root, R.
mimosoides or C. palala root and Microporus xanthopus decoction. This is to
ensure full recovery and provide energy to the mother. Interestingly, Jakun
women also used one species of fungi which is M. xanthopus.
310
Table 1. Medicinal plants used for women healthcare in Jakun community.
Botanical
information
Parts
used
Methods
of Application
preparation
1.
L. pumila Benth.
Kacip fatimah
Myrsinaceae
Root
Decoction
water
2.
C. palala
Pengesep
Connaraceae
R.
mimosoides
(Vahl.) Planch
Pengesep
Connaraceae
M. xanthopus
Kulat kelentit kering
(3K)
Polyporaceae
Root
Decoction
water
Root
Decoction
water
Fruiting
body
Helminthostachys
zeylanica (L.) Hook
Paku tunjuk langit
Ophioglossaceae
Rennellia
elliptica
Korth.
Segindu
Rubiaceae
Mapania sp.
Pandan
serapat
Cyperaceae
Penggugur
Root
Decoction in
water
(decoction
with L. pumila
and C. palala)
Decoction in
water
Raw
Ixonanthes icosandra
Jack
Pepagar
Ixonanthaceae
10. Cinnamomum sp.
Teja lawang
11. Melastoma sp.
Sekenduduk putih
Melastomataceae
Root
3.
4.
5.
6.
7.
8.
9.
Flower
Ways of
administration
in Post-partum recovery Oral : drink
(inducing
uterus
shrinking); Fertility &
health tonic
(inducing
uterus
shrinking); Health tonic
in Post-partum recovery; Oral : drink
Health tonic
Post-partum recovery Oral : drink
(inducing
uterus
shrinking)
Post-partum recovery;
For beauty purposes
Oral : drink
Topical :
applied to
skin
Oral : drink
Root
Decoction
water
in Facilitate childbirth
Root
Decoction
water
Root
Decoction
water
Decoction
water
(inducing
uterus
shrinking)
in Abortifacient
Oral : drink
Root
Leaves
Flower
Decoction
water
Decoction
water
Raw
311
in Induce fertility
Oral : drink
in Post-partum recovery
Oral : drink
in Post-partum recovery;
Internal body pain
Oral : drink
Eaten raw
CONCLUSION
The results of our preliminary study shows that the women in Jakun
community still rely on the medicinal plants to treat women’s disorder. Future
investigation and proper documentation need to be done in order to preserve
their knowledge from erosion.
ACKNOWLEDGEMENT
The authors wish to thank the Johor National Park Corporation for their
assistance during fieldwork and Jakun community in Kampung Peta, EndauRompin for their time, effort and willingness to share their knowledge with us.
REFERENCES
D’Avigdor, Elizabeth, Hans Wohlmuth, Zemede Asfaw, & Tesfaye Awas. 2014.
The Current Status of Knowledge of Herbal Medicine and Medicinal
Plants in Fiche , Ethiopia. Journal of Ethnobiology and Ethnomedicine
10 (38): 1–32.
de Boer, H. & Crystle Cotingting. 2014. Medicinal Plants for Women’s
Healthcare in Southeast Asia : A Meta-Analysis of Their
Traditional
Use, Chemical Constituents, and Pharmacology. Journal of
Ethnopharmacology 151 (2): 747–67.
Pfeiffer, J.M. & Butz, R.J. 2005. Assessing Cultural and Ecological Variation in
Ethnobiological Research : the Importance Of
Gender. Journal of
Ethnobiology 25: 240–278.
Taylor, C. E. & Wong, K. M. 1987. Some Aspects of Herbal Medicine Among the
Orang Hulu Community of Kampung Peta, Johore, Malaysia. Malaysian
Heritage & Scientific Expedition: Endau Rompin. The Malayan Nature
Journal, 41 (2 &3): 317─328.
312
KAEMPFERIA L.: HERBS
ZINGIBERACEAE SPECIES
AND
ORNAMENTAL
POTENTIAL
OF
R Izlamira1, MA Zulkhairi2, MZ Nurin Izzati2 & B Suryanti2
1
Programme of Genetic Resources and Germplasm Conservation Management
(GB1), 2Genebank and Seed Centre (GB), MARDI Jerangau, 21820 Ajil,
Terengganu
Tel: 013-2752142 Fax: 09-838 4208 E-mail: [email protected]
ABSTRACT
Kaempferia is a medium-sized genus of small herbaceous in the Zingiberaceae
family. This genus is known for its outstanding beautiful foliage and flower.
Seven species were collected during the study. There are K. galanga L., K.
pulchra Ridl., K. angustifolia Roscoe, K. rotunda L., K. parviflora Wall. ex Baker,
K. gilbertii W. Bull and K. elegens (Wall.) Baker. Most of the Kaempferia species
are generally used as medicinal properties and ornamental except for K.
gilbertii and K. elegens. These two species is less known for its medicinal value,
however showing a great potential as ornamental. This is based on the
beautiful morphological characteristics of their foliage which usually variegated
on the upper surface. All the accessions of collected species were conserved in
MARDI Jerangau germplasm. The genetic diversity serves as the gene pool that
will provide important genes for the future study in the broader scope.
Keywords: Kaempferia, herbal plants, ornamental
INTRODUCTION
The genus Kaempferia L. comprises about 70 species geographically distributed
from tropical Africa to India and throughout Southeast Asia (Kumar et al.
2013). In Peninsular Malaysia, several taxa such as Kaempferia. galanga, K.
pulchra, K. parviflora and K. rotunda are known locally for their medicinal
properties. The rhizomes of K. galanga are carmative and useful for skin
problems, sore eyes, tonics, inflammation, childbirth, cough, sore throat and
fever (Idha & Helmy 2011). The leaves are also being used in traditional
medicine to treat swelling, headache and rheumatism. K. parviflora is
traditionally used for health promoting, stimulating and vitalizing. It is also
used in several treatments such as dysentery, impotence, constriction and colic
disorders. K. rotunda is widely used in traditional medicine as a wound healing
agent (Syed et al. 2013).
313
Other than its medicinal values, Kaempferia species also have a great
potential as ornamental or landscape plant. It is reported to be one of the most
common genera used as an ornamental other than Curcuma and Globba
(Kuehny 2001, Kuehny et al. 2002). This genus is known for its outstanding
beautiful foliage and flower with most of them having a variegated silver to
purple feather pattern on the upper surface of the leaf. K. pulchra, a wild
species which discovered in the limestone hills of Langkawi Island has been
successfully brought into cultivation, growing in pots, on the ground in shaded
areas and gardens (Larsen et al. 1999). Several other species, including K.
galanga and K. elegens are also often used as a pot plant and decorative
garden plant (Picheansoonthon & Koonterm 2008). The morphological
characteristics of Kaempferia such as size, colour of flower and unique foliage
make this genus gaining increase recognition as an attractive ornamental
species.
The live plants of Kaempferia accession were collected from wild and home
garden in Peninsular Malaysia. The Kaempferia species available in the field as
well as their uses were recorded. The collected plants are successfully
conserved in MARDI Jerangau germplasm for further references. Literature
review was also collected from different sources such as books, journal,
internet database and others to gather information on the potential uses of
Kaempferia species.
Seven species of Kaempferia were recognized from a total number of 34
collected accessions during the study. There are K. galanga, K. pulchra, K.
angustifolia, K. rotunda, K. parviflora, K. gilbertii and K. Elegens( Figure 1).
314
a
d
b
e
c
f
g
Figure 1. (a) K. galanga, (b) K. pulchra, (c) K. angustifolia, (d) K. rotunda, (e) K.
parviflora, (f) K. gilbertii and (g) K. elegens
K. galanga L.
Local name: Cekur, cekur Jawa, cengkur or kencur
Distinguished characters: Rhizome strongly aromatic; leaves green, often
horizontal and flat on the ground, lamina elliptic to broad elliptic; inflorescence
enclosed in the two leaf-sheaths; flower white with violet bands in the basal
half.
Uses: ‘Ulam’, medicinal and ornamental; growing as pot plants, the leaves and
rhizomes are used for food and medicine, the rhizomes are an ingredient of
post partum medicine and to treat common cold.
K. pulchra Ridl.
Local name: Cekur batik, cekur hitam or cekur mas
Distinguished characters: Leaves broad elliptic, dark purple to dark green with
or without greyish spot; sheath closely imbricating and enclosing the
inflorescence; flower lilac except for a small white at the base.
Uses: ‘Ulam', medicinal and ornamental; planted as garden ground cover and
pot herbs.
K. angustifolia Roscoe
Local name: Cekur rumput or kunci pepet
Distinguished characters: Leaves erect, green, elliptic-oblong to lanceolate;
inflorescence borne in the two innermost leaf-sheath; flower lilac with a purple
blotch at the centre.
315
Uses: Spiritual and medicinal purposes, the small roots and tubers have
astringent properties and are used in the treatment of cough, dysentery and
diarrhoea.
K. rotunda L.
Local name: Kunyit putih or temu putih
Distinguished characters: Leaves oblong lanceolate, mottled green above,
purple beneath; Inflorescence appearing before the leaves. This genus easily
recognize as the flower consist of four lobes.
Uses: ‘Ulam', medicinal and ornamental; good for healing fresh wounds, leaves
and rhizomes are eaten fresh or cooked and used in cosmetic powder and as a
food flavouring agent.
K. parviflora Wall. ex Baker
Local name: Kunyit hitam
Distinguished characters: Rhizome dark purple or greyish to black; leaves
ovate or elliptic, green with reddish margin; flower purple, darker in the middle
with the emarginated apex.
Uses: Its black rhizomes are high medicinal value and claimed to possess a
potent male aphrodisiac effect.
K. gilbertii W. Bull
Local name: Unknown
Distinguished characters: Leaves are striped at the margins with white
variegation.
Uses: Growing for its beautiful foliage as potting ornamental plant.
K. elegens (Wall.) Baker
Local name: ‘Peacock ginger'
Distinguished characters: Rhizomes yellow; leaves elliptic, spotted strongly
with silver and dark green on upper surface.
Uses: Garden decoration, pot plant, ground cover and indoor plant.
The result shows that Kaempferia species are an important ornamental
plant apart from its high medicinal values. The morphological characteristics of
each species make this genus very well known for its outstanding beautiful
foliage and flower. Conservation of Kaempferia diversity is needed to protect
such potentially useful economic plant for present use and future study in the
broadest scope.
316
CONCLUSION
Other than its medicinal value Kaempferia species also have a great potential
as ornamental or landscape plant.
REFERENCES
Kumar, K.M., Asish, G.R., Sabu, M. & Balachandran, I. (2013). Significance of
gingers (Zingiberaceae) in Indian System of
Medicine ─ Ayurveda: An
overview. Anc Sci Life: 32(4): 253–61.
Idha, K. & Helmy, Y. (2011). Phospholipid complex as a carrier of Kaempferia
galanga rhizome extract to improve its analgesic activity. International
Journal of Pharmacy and
Pharmaceutical Sciences: (3): 44–46.
Syed, A.I., Saroj, K.R., Niranjan, S., Uma, S.S. & Ranju, S. (2013).
Wound
healing activity of Kaempferia rotunda Linn leaf
extract. International
Journal of current microbiology and Applied Sciences: 2(12): 7478.
Kuehny, J.S. (2001). Potting ornamental ginger. Potted crops. Greenhouse
Product News. 30–32.
Kuehny, J.S., Sarmiento, M.J. & Branch, P.C. (2002). Cultural studies
in
ornamental ginger. Trends in New Crops and New Uses: 477–479.
Picheansoonthon, C. & Koonterm, S. (2008). Notes on the genus Kaempferia L.
(Zingiberaceae) in Thailand. Journal of Thai
Traditional & Alternative
Medicine: 6 (1): 27–51.
Larsen, K., Ibrahim, H., Khaw, S.H. & Saw, L.G. (1999). Gingers of Peninsular
Malaysia and Singapore. Natural History Publications (Borneo), Kota
Kinabalu.
317
DISTRIBUTION OF HERBS CULTIVATION IN PENINSULAR MALAYSIA
A Nur Syazni, AR Rohana, AB Ariff Fahmi, Z Nur Fazreen, S Siti Zubaidah & M
Marzalina
Forest Research Institute Malaysia (FRIM), 52109 Kepong, Selangor Darul
Ehsan, Malaysia
ABSTRACT
In general, herbs are plants that are used for medicinal, food, flavoring and
perfume purposes. Recently, there is an increse on the interest of herbal
products in Malaysia. Under the National Key Economic Areas (NKEA), high
value herbal products is listed under Entry Point Project 1 (EPP 1) focuses in
improving quality and marketing effort. This includes in ensuring sufficient
supply of raw material to fulfil market needs. However, there are issues on lack
of information on location of the herbs cultivation in this country. This paper
highlights distribution of herbs cultivation in Peninsular Malaysia. A census was
carried out from September 2014 to February 2015 to identify location of
herbs cultivators along the roads in Peninsular Malaysia. The information was
gathered based on questionnaire and through face-to-face interview. A total of
462 cultivators were identified in Peninsular Malaysia. Most of the herbs were
cultivated in Pahang, Selangor, Johor and Perak with a total number of 110, 83,
69 and 67 cultivators respectively. In Pahang the most cultivated herb was
Hibiscus sabdariffa (roselle), meanwhile in Selangor, Johor and Perak the
famous herb cultivated were Piper betle (sireh), Aquilaria malaccensis (karas)
and Citrus aurantifolia (limau nipis), respectively. The findings will be useful as
baseline information to further strengthen the industry by identifying the
stakeholders and market structure.
Keywords: herbs cultivators, distribution, herbal industry, raw materials,
Peninsular Malaysia
INTRODUCTION
Herbs are known as plants, seeds, or any plant parts that are used as medicine,
flavoring and perfume purposes and even some are taken raw. According to
Sahri (2012), herbs also have been recognized as an alternative medicine and
economical resources. Recently there has been a shift in universal trend from
synthetic to herbal medicine, which can be said as ‘Return to Nature’ (Sharma
et al. 2008). The herbs became famous when there are perceptions that herbal
318
products have fewer or less harmful side effects compared to synthetic or nonherbal products. Most of the information on usage as well as benefits of herbs
are only based on traditional knowledge from various ethnics inherited from
the ancestors and only few are documented. Malaysia has been put as having a
vast potential size in terms of herbal based market with the diversity of genetic
resources, excellent tropical climate, increasing research and development
interest, increasing demand for natural products and indigenous knowledge
(Ibrahim 2006).
In current situation, many products had been made based on these
local herbs and consumed as health supplements and not forgetting to prevent
diseases. In order for the herbal industry to go upscale, a comprehensive
research and clinical trial are required. This is critical not only before the
production but also to monitor the quality and safety of the products in the
market. Scientific evidence is needed to prove the quality of local herbs to
obtain confidence from the other countries towards our products in Malaysia
and fulfill their demand. Under the National Key Economic Areas (NKEA), high
value herbal products is listed under EPP 1 that looks at improving the quality
and efforts in marketing local herbal products. However, there is not much
information on the location of the herbs cultivation and raw material. This
study was conducted in order to identify the distribution of herbs cultivation in
Peninsular Malaysia.
Study Area
This study was conducted across the Peninsular Malaysia which included 12
states. The target respondents for this census study were all herbal cultivators
found in the study area.
Data collection
Census was done based on a set of questionnaire developed to collect the
information. The questionnaire contained 19 questions consisted of
background of the company, name of the owner, address, contact number,
gender, ethnicity and nature of business.
Data Analysis
The data was analyzed according to number of cultivators by states and herbal
species that were cultivated by states. Further analysis was done using Chi319
square test to understand the correlation between the five most cultivated
herbal species and ethnicity of the herbal cultivators.
Table 1 showed the number of herb cultivators according to states in
Peninsular Malaysia. Pahang recorded the highest number of herbs cultivators
with 110 cultivators (23.8%). No cultivators were found in the Federal
Territory. There were only 4 herbs cultivators discovered in Perlis.
Table 1. Number of cultivators according to states in Peninsular Malaysia
State
Johor
Kedah
Kelantan
Melaka
Negeri Sembilan
Pahang
Perak
Perlis
Pulau Pinang
Selangor
Terengganu
Federal Territory
Total
Number of cultivators (%)
69 (14.9)
19 (4.1)
29 (6.3)
27 (5.8)
28 (6.1)
110 (23.8)
67 (14.5)
4 (0.87)
19 (4.1)
83 (18.0)
7 (1.5)
0 (0)
462 (100)
In Pahang, the most famous cultivated herb species cultivated was
Hibiscus sabdariffa (Roselle) with 52 cultivators, meanwhile in Selangor the
most famous cultivated herb species was Piper betle (sireh) with 65 cultivators
(Table 2). According to Table 2, the highest number of herb species cultivated
in Peninsular Malaysia was H. sabdariffa with 88 cultivations compared to
other species such as Aquilaria malaccensis (karas) and P. betle with only 70
and 80 cultivators, respectively.
320
Table 2. Types of herbs cultivated in states across Peninsular Malaysia
Species/ States
JHR
KDH
KEL
MEL
N.S
PHG
PRK
PER
P.P
SEL
TGG
Total
Hibiscus sabdariffa (Roselle)
Orthosiphon stamineus (Misai kucing)
Eurycoma longifolia (Tongkat ali)
Clinacanthus nutans (Belalai gajah)
Ficus deltoidea (Mas cotek)
Labisia pumila (Kacip fatimah)
Momordica charantia (Peria katak)
Piper betle (Sireh)
Cymbopogon nardus (Serai wangi)
Morinda citrifolia (Mengkudu)
Phaleria macrocarpa (Mahkota dewa)
Senna alata (Gelenggang)
Centella asiatica (Pegaga)
Aquilaria malaccensis (Karas)
Citrofortunella japonica (Limau kasturi)
Citrus aurantifolia (Limau nipis)
Andrographis paniculata (Hempedu bumi)
Zingiber officinale (Halia)
Melastoma malabathricum (Senduduk)
Melastoma candidum (Senduduk putih)
Moringa oleifera (Merunggai)
Phyllantus niruri (Dukung anak)
Others
15
4
4
5
7
2
1
5
4
2
4
1
2
17
0
1
1
0
1
1
0
0
18
2
1
0
0
1
1
1
0
1
0
1
3
0
1
0
1
0
0
1
1
0
0
8
1
6
8
4
1
1
0
1
3
0
5
1
0
4
0
0
0
1
1
0
0
0
7
2
2
0
0
0
0
1
2
2
0
0
0
1
5
6
4
0
0
1
0
0
0
4
0
5
3
3
4
0
0
1
0
0
3
1
0
15
0
0
2
0
0
0
1
2
4
52
2
9
5
2
0
0
1
2
1
2
2
0
12
5
14
1
5
0
0
0
0
14
6
8
1
5
1
0
2
3
1
1
5
1
1
13
5
28
0
0
0
0
0
0
13
2
1
0
1
0
0
0
0
0
0
0
0
2
0
1
1
0
0
0
0
0
0
0
6
7
1
5
1
1
1
1
1
0
0
6
1
0
0
0
2
2
0
1
2
1
5
0
2
0
1
6
0
1
65
0
0
0
0
2
3
0
1
0
0
0
0
0
0
7
2
0
1
0
0
0
0
1
0
0
1
0
0
0
0
1
1
0
0
0
0
0
1
88
38
27
29
23
5
7
80
14
4
21
15
9
70
17
51
7
8
4
3
3
3
81
Source: FRIM’s survey, 2014-2015.
321
Table 3 shows the correlation between the number of respondents
based on herbal species and ethnicity. Based on the table, majority of the
cultivators were Malay. According to reports on socio-economic status of
farmers in North West Selangor area by Alam et al. (2010), variations in factors
such as education, politic, wealth, health status, accession to technology,
formal and informal capital were responsible for the variations in socioeconomic characteristics of farmers.
It was noted that P. betle was mostly cultivated or planted herb by the
Indian community and used as medicinal plants. It was also believed that the
species is related to symbol of etiquette and civility among the community
(Kumar 1999).
Table 3. Number of cultivators based on ethnicity and selected species
Species
Hibiscus sabdariffa
(Roselle)
Piper betle
(Sireh)
Aquilaria malaccensis
(Karas)
Orthosiphon stamineus
(Misai kucing)
Clinacanthus nutans
(Belalai gajah)
Malay
Chinese
India
84
3
1
11
0
52
Ethnicity
Other
Foreigner
Unknown
0
0
0
68
0
0
1
9
3
0
0
6
31
5
1
0
0
1
25
3
0
0
0
1
CONCLUSION
As a conclusion, H. sabdariffa was the most cultivated herbal species in
Peninsular Malaysia, which is in the state of Pahang. We also came to
conclusion that most cultivators of this species were from the Malay
community. Further studies must be carried out to identify the adaptive
capacity of individuals or communities based on internal characteristics that
were responsible for these variations.
ACKNOWLEDGEMENT
We would like to acknowledge FRIM’s Director General, Y.Bhg. Dato’ Dr. Abd
Latif Mohmod for his support, motivation and assistance. We also thank MOA
for providing the financial support through NRGS fund. Our deepest
appreciation to Herbal Development Division for their support, collaborating
agencies, DoA, MARDI, IMR, FAMA, FELCRA, FELDA, LTKN, KESEDAR, KETENGAH
322
and RISDA for their cooperation, and to the research team members and all
herbal cultivators who were involved in this study.
REFERENCES
Alam, M.M., Siwar, C., Murad, M.W., Molla, R.I., and Mohd Ekhwan. T. (2010).
Socioeconomic Profile of Farmer in Malaysia: Study on Integrated
Agricultural Development Area in North-West Selangor, Agricultural
Economics and Rural Development, Vol. 7(2), pp. 249-265. Available at
<http://www.ipe.ro/RePEc/iag/iag_pdf/AERD1013_249-265.pdf > (ISSN
1841-0375; Publisher- Institute for Agricultural Economics, National
Institute of Economic Research, Romania; India; Indexed in EconLit,
RePEc)
Ibrahim J. (2006). The scientific values of Malaysian herbal products. Malaysian
Journal of Health Science, 4(1), 59─70.
Kumar, N. (1999). Betelvine (Piper Betle L.) cultivation: a unique case of plant
establishment under anthropogenically regulated microclimatic
conditions. Indian Journal of History of Science, 34(1)
Rohana, A.R, Nur Fazreen Z, Ariff Fahmi Ab, Nur Syazni A, Siti Zubaidah S, Lim
Hf & Marzalina M. 2015. Value chain study for holistic development of
herbal industry. Pp. 211–219 in Proceeding 3rd International
conference on Rural Development and Entrepreneurship (ICORE) 2015,
9–11 May, Hebei Province, China.
Rohana, A.R., Nur Fazreen, Z., Ariff Fahmi, A.B., Nur Syazni, A., Siti Zubaidah, S.,
Lim, H.F., Mohd Shahidan, M.A., Rosniza, R., Marzalina, M. & Abd Latif,
M. 2015. Directory of Herbal Cultivators in Peninsular Malaysia, Forest
Research Insitute Malaysia.
Sahri, Z. (2012). Malaysia indigenous herbs knowledge representation (Doctoral
dissertation, Universiti Teknologi MARA).
Sharma, A., Shanker, C., Tyagi, L. K., Singh, M., & Rao, C. V. (2008). Herbal
medicine for market potential in India: an overview. Acad J Plant
Sci, 1(2), 26─36.
323
HALAL HERBAL PRODUCTS INTEGRITY RISK THROUGH SUPPLY CHAIN: A
CONCEPTUAL STUDY
R Salini Devi1 & K Nitty Hirawaty1,2
1
Halal Products Research Institute, 2Department of Agribusiness and
Information Systems, Faculty of Agriculture, Universiti Putra Malaysia, 43400
UPM Serdang, Selangor
ABSTRACT
There are thousands of herbal products and supplements available in the
market as a nutritional boost particularly for families who strive to maintain a
healthy, halal lifestyle. An increased awareness on the breadth of herbal
labelled as halal is crucial, as consumers globally strive for halal consumption.
The length and complexity of herbal supply chains raise questions on the
authenticity of herbal products labelled as halal and halal integrity of the
supply chain. The risks involved in ensuring product integrity as it moves
through multiple tiers of suppliers, further aggravates the uncertainty. Thus, it
is very challenging for the manufacturers to reduce risks and maintain the
integrity of halal herbal products. As the halal market is huge, demand for
halal herbal products is expected to grow bigger in the future, thus the risks
must be given serious attention to maintain the integrity of halal herbal
products in providing halalan toyyiban products for the needs and well-being
of the consumers. Thus, this conceptual paper will investigate the risks in halal
herbal products supply chain. A quantitative study will be carried out to obtain
information from supply chain players including herbal-based manufacturers
listed by FRIM and JAKIM. Data will be analysed using descriptive statistics,
regression analysis, chi-square test, factor analysis, correlation analysis and
CFA. The results are expected to reveal that halal integrity risk such as
production risk, raw materials risk, food security risk, outsourcing practices
risk, service risk, and logistics risks influence halal integrity along the herbal
supply chain.
Keywords: herbal products, risks, halal supply chain, integrity, halalan toyyiban
324
INTRODUCTION
Malaysia is well known for its rich natural resources and dense forests.
According to MARDI (2006), the rich flora of Malaysia includes 15,000 known
plant species, of which 2,000 species are used for its medicinal value. This
plentiful supply has contributed to the development of natural products, as
ingredients for food and drinks, cosmetics, and natural health & dietary
supplements. The most popular herb species used by the Malaysian herbal
industry for development of natural products are belalai gajah, dukung anak,
gelenggang, halia, hempedu bumi, kacip fatimah, karas, makhota dewa, mas
cotek, mengkudu, merunggai, misai kucing, pegaga, peria katak, rozel,
sambung nyawa, senduduk putih, serai wangi, sirih, and tongkat ali (FRIM
2015). The increasing health consciousness among Malaysians, coupled with an
increase in expendable income, higher levels of education, and population
growth has increased the demand for natural foods and drinks, natural
cosmetics as well as health supplements. Recently, collagen has gained the
attention of Malaysians, as an ingredient in food and drink products, cosmetics,
medical, and health supplements. However, it is not blindly accepted by the
Muslim consumers. Collagen extracted from ungulates (cattle, pig, and sheep)
are not easily marketable in Malaysia, due to religious obligations of halalan
toyyiban (Mohd. Rizal & Adham 2010). Even though medicines are exempted
from halal food regulations, a prescription medicine encapsulated in a
prohibited gelatine capsule made of porcine gelatine is still considered haram
and unacceptable by Muslims. Due to this, efforts are ongoing in research into
possibilities in halal pharmaceuticals as seen in the case of halal vaccines, as
well as research on nutraceuticals focusing on high-value herbal products like
tongkat ali, misai kucing, hempedu bumi, dukung anak, and kacip fatimah.
HALAL DEFINITION
Halal originates from an Arabic term which means allowed, permissible, lawful
or legal. This term specified that goods or services deemed not harmful and
emphasised as safe to be consumed, by the Syariah law, are permissible or
allowable. The opposite is haram or non-halal which means forbidden,
unlawful or illegal (Tieman 2011). Thereby, according to the Islamic principles
some things must be avoided (Ibn Abd al-Barr 2000). In general, halal and
haram has become important elements for consideration in a Muslim’s life.
The Muslims want assurance that the products they consume, not only follow
the Islamic law , but must be toyyib, which means pure and good (World Halal
Forum 2009; Anas et al. 2010).
As pointed out in the Syariah guidelines, any food, drink, cosmetics and
health supplements must not be najs; or contain things that are najs in or for
325
their processing; or processed using equipment or utensils that are najs; or
come into contact at any time during their preparation, processing and storing.
Najs includes meat from inappropriate slaughtered animals, refuse, faeces,
alcoholic beverages, carrion, as well as pork and all of its by products (Sabiq
2008). Furthermore, according to JAKIM (2009), activities of supply chain such
as processes, manufacturing and packaging, receiving, handling, storing and
delivery of products must be verified and certified as halalan toyyiban by
authorized persons.
HALAL INTEGRITY
Nowadays, consumers are concern about food safety, quality, hygiene and
humane approach along the entire production supply chain. Therefore,
integrity of halal food supply chains has become an important subject matter
(Zailani et al. 2010; Lam & Alhashmi 2008). A number of reasons influence the
integrity of halal food chains. One of the main causes is the length and
complexity of food supply chains that involve lots of handling as it is moved
across great distances before reaching the end consumers. The task of ensuring
halal integrity along such chains is a humongous task to manage and monitor.
The reality of this situation has raised questions on the credibility of halal
status of food products. According to Lodhi (2009); Khan (2009); Tieman (2011)
halal integrity is one of the main contributors to developing a well trusted halal
food supply chain. The halal integrity of a product, particularly herbal-based
products will not be compromised as it moves from upstream to downstream
along the herbal supply chain, which starts from the procurement of herbal
raw materials till the finished products reach the consumers. Previous
literature has revealed that all players in the supply chain should take the
responsibility without leaving the huge tasks to a single player (Mohd Hafiz et
al. 2013; Melatu Samsi et al. 2011). Past studies on halal indicated that the
halal integrity of a product is only intact when the product is still in the custody
of an organization (Jaafar et al. 2011). Once a product is moved along the
supply chain, the quality assurance depends on the next player who handles
that particular product. Thus, the integrity of a product cannot be guaranteed if
the other players in the supply chain do not practice similar halal concepts. The
halal herbal products supply chain has similar fundamental principles as other
food supply chains that is, “from farm to fork” concept. The long and complex
herbal-based product supply chains still remain an obstacle to ensure halal
integrity, due to the involvement of many players including suppliers and
producers/manufacturers. Therefore, there is also a risk to food integrity along
the supply chain. Past literatures revealed six risks associated with halal
integrity along supply chains; namely production risks, raw material risks, food
security risks, outsourcing practices, service risk, and logistics risk. Table 1
shows the six types of risks correlated with halal integrity.
326
Table 1. Halal Integrity Risks
Types of
Risks
Production
Raw
Materials
Food
Security
Services
Outsourcing
Logistics
Explanation
Production is known as the most important stage in determining the
quality of the food for the customer. Along the long and complex halal
food supply chain, the most significant risk at the production stage is
the contamination from non-halal elements used in halal certified
ingredients. It is a must for manufacturers to obtain halal certification
which covers the ingredients used for production. Besides, Talib and
Johan (2012) mentioned that contamination from the equipment may
affect halal integrity.
Very few countries aside from Malaysia implement halal food
certification and standards through the government’s collaboration.
Due to this, the integrity of imported raw materials cannot be assured.
Tse and Tan (2011) supported that there is high risk to halal integrity
especially when there is low traceability of material origins. It is
understandable that the confirmation of halal certification practices by
suppliers is only set up at the very early stage of appointment.
As the process of supplier selection becomes inflexible and strict, firms
have limited options to leverage the potential source available in the
market. As a result of this, the local halal food industry directs the firm
to source from the international market. Other than that, halal food
security can be breached due to unexpected situations such as unusual
and/or seasonal demands, product shortages, poor management, and
price fluctuations of certain raw materials in the small local market. This
often leads to high risks for halal integrity (Mohd Helmi et al. 2013).
According to Wildes (2005), the most critical issue in an effective service
delivery system is the chance of human errors. From the perspective of
halal food production, a specialized and dedicated group of workers
should handle the production in order to minimize the probabilities of
human error. Generally, halal integrity concentrates on the risk of
human error and cross contamination particularly in a restaurant
setting (Talib & Johan 2012).
Current trends show that firms intend to outsource various processes
such as logistics, warehousing, packaging, marketing and other related
activities in order to lower their production costs. Their main goal is to
focus on core business within the firms and outsource other activities to
other service providers. The outsourcing makes the supply chain longer
and difficult to keep track of along the chain. As a result, firms often
lose control and lead to high integrity risks (Lyless et al. 2008).
The halal guidelines clearly state that halal products should not be
contaminated with non-halal products. Jaafar et al. (2011) revealed in
his study that transportation vehicles need to be cleaned and
consignments properly segregated in order to ensure that the
requirements are observed and the logistics dedicated to the halal
product shipment. This has become a major challenge for the industry
as there are very few halal logistics providers to cater to this demand.
327
It is inarguable that ensuring the halal integrity of herbal-based
products remains the biggest challenge in the halal herbal industry. The halal
food market including herbal-based market is a huge and lucrative business. It
is estimated to grow significantly in the next decade. Thus, the herbal-based
products supply chain players must ensure that there is no contamination,
intentionally or unintentionally in producing the products. More importantly,
halal integrity has become one of the key successes for the entire halal
industry, and without halal integrity, we as consumers would be living in a
maze of 'false integrity'. Thus, this concept paper aims to study on the
relationship between risks and halal integrity of herbs and herbal products
along the herbal supply chain.
METHODOLOGY
Quantitative technique will be applied to obtain required information from
herbal based product manufacturers listed by FRIM and JAKIM. The sources of
data for this study will be both primary and secondary data. The respondents
will be given questionnaire booklets to be answered. The questionnaire will
consist of both closed-ended and open-ended questions. Most of the questions
will be structured questions, followed by dichotomous, multiple choices, and
Likert scale statements. To measure the factors influencing halal integrity along
the supply chain, a 5-point Likert scale will be used. The respondents will be
asked to indicate whether he/she agrees or disagrees with each statement. The
questionnaire will be divided into four sections. The first section is designed to
obtain socio-demographic details of the respondents. The second section is
designed to obtain information on how manufacturers practiced halal
principles, the third section is designed to determine the factors influencing
halal integrity in supply chain and finally the last section is developed to
investigate the halal food integrity risks through supply chain. In this study, the
sample will be designed using a probability sampling procedures which is
simple random sampling. The population of the research is the herbal based
product manufacturers in Malaysia. The analysis methods of descriptive
statistics, correlation analysis, factor analysis, regression analysis, chi-square
test, and confirmatory factor analysis (CFA) will be used to analyze the data.
CONCLUSION
As a matter of fact, it is very challenging for firms to ensure the integrity of
halal products. However, as the halal market is huge and profitable, and the
demand for halal food products is expected to grow even bigger in the near
328
future, the risks must be given serious attention in order to ensure the needs
and well-being of the halal food consumers. Based on the findings, it is hoped
that the major players in the halal industry especially the suppliers, producers,
logistic service providers, wholesalers, traders, and government will give higher
consideration to the factors that influence the integrity of halal herbal food
supply chain. Besides that, it is also hoped that the outcome from this study
could improve and enhance competitiveness of the Malaysian halal herbal
industry.
REFERENCES
Halal Malaysia-Jabatan Kemajuan Islam Malaysia (JAKIM) 2009. Takrifan Halal
[in Malay].
http://frim.gov.my. Access on 17th September 2015.
http://mardi.gov.my. Access on 18th September 2015.
Jaafar, H.S., Endut, I.R., Faisol, N. & Omar,E.N. (2011). Innovation in Logistics
Services - Halal Logistic. In Proceedings of the 16th International
Symposium on Logistics (ISL), Berlin, Germany, no. 34665, 844–851.
Khan, N. (2009) Special Report: Halal Logistics.
Lam, Y. & Alhashmi, S.M. (2008). Simulation of Halal Food Supply Chain with
Certification System: A Multi-agent System Approach. PRIMA ’08:
Proceedings of the 11th Pacific Rim International Conference on Multiagents: Intelligent Agents and Multi-agent System
Lodhi, A.-u.-H. (2009). Understanding Halal Food Supply Chain, London: HFRC
UK Ltd
Lyles, M.A., & Flynn, B.B. & Frohlich, M.T. (2008). All Supply Chains Don’t Flow
Through: Understanding Supply Chain Issues in Product Recalls.
Management and Organization Review, 4(2), 167–182
Melatu Samsi, S.Z., Tasnim, R. & Ibrahim, O. (2011). Stakeholders' Role for an
Efficient Traceability System in Halal Industry Supply Chain, in Annual
International Conference on Enterprise Resource Planning + Supply
Chain Management (ERP + SCM 2011): Penang, Malaysia.
329
Mohd Hafiz, Z., Marhani, M.A. & Mohamed Syazwan, Ab.T. (2014). Conceptual
Framework on Halal Food Supply Chain Integrity Enhancement.
Procedia - Social and Behavioral Sciences, (121), 58 – 67.
Mohd Helmi, A., Kim, H.T. & Zafir, M.M. (2013). Mitigating Halal Food Integrity
Risk through Supply Chain Integration. Asia Pacific Industrial
Engineering and Management System.
Sabiq, S. (2008). Fiqih Sunnah. Jakarta: Pena Pundi Aksara, Page: 25–65. [in
Malay].
Talib. M & Johan, M. (2012). Issues in Halal Packaging: A Conceptual Paper.
International Business and Management, 5(2), 94–98
Tieman, M. (2011). The Application of Halal in Supply Chain Management: InDepth Interviews. Journal of Islamic Marketing, 2(2), 186-195.
Tse, Y.K & Tan, K.H. (2011). Managing Product Quality Risk in a Multi-tier Global
Supply Chain. International Journal of Production Research, 49(1),139–
158
Wildes, V.J. (2005). Stigma in Food Service Work: How it Affects Restaurant
Servers’ Intention to Stay in the Business or Recommend a Job to
Another, Tourism and Hospitality Research, 5(3), 213–233
Zailani, S., Arrifin, Z., Abd Wahid, N., Othman, R. & Fernando, Y. (2010). Halal
Traceability and Halal Tracking Systems in Strengthening Halal Food
Supply Chains for Food Industry in Malaysia (a review). Journal of Food
Technology, 8(3), 74─81.
330
CONSUMER PREFERENCES IN SELECTING HERBAL PRODUCTS
S Siti Zubaidah, AR Rohana, AB Ariff Fahmi & A Nur Syazni
ABSTRACT
Malaysia has identified that herbal industry could help in generating gross
national income (GNI). Under National Key Economic Area (NKEA), in
transitioning from agriculture to agribusiness, the first entry point project is
unlocking value from Malaysia’s biodiversity through high-value herbal
product. One of the efforts to produce high-value herbal products is the
industry need to properly strategize their production based on actual
consumer preference. In order to understand consumers’ behavior on herbs
and herbal based products, a study on Consumer Preference and Demand
towards Herbal Based Product in Peninsular Malaysia was conducted in 2013.
The survey was conducted by using stratified random sampling based on
population in each district of Peninsular Malaysia. A total of 4,452 respondents
were interviewed using structured personal interview. The result from this
study highlights status on the use of herbal based products among the
respondents and their criteria in selecting herbal products. The findings
showed that 73% of respondents were currently using herbal based products.
A total of 2,499 respondents (76%) agreed that products with certificate from
Ministry of Health (MoH) are the most important aspect in herbal based
product selection. Aside from MoH certificate, Good Manufacturing Practice
(GMP) certificate was also an important concern of 72% of the respondents
concern. This study showed that the industry, especially manufacturers should
produce herbal based product with good quality and safety. Related agencies
should also help the manufacturer acknowledged to better understand the
procedure to get certificates.
Keywords: herbal industry, consumer preference, market driven, Peninsular
Malaysia
INTRODUCTION
Import value for herbal based products such as plant based, ginseng root,
additional food, skincare products and pharmaceutical in 2012 was USD 402
million (Rohana et al. 2013). This indicated that demand for herbal based
products among Malaysians is quite high. Rohana et al. (2014) reported that
45% of the herbs and herbal products consumers in Malaysia used herb in
traditional way while 46% of them used contemporary value-added products.
331
In order to produce high-value herbal products that meet consumer demand,
production based on the actual consumer preferences are needed.
The respondents for this study were indentified by stratified random sampling
based on population in each Majlis Perbandaran with consideration on the
gender and age to represent the whole Peninsular Malaysia. A total of 4,452
respondents were interviewed via personal interview using structured
questionnaire.
The questionnaire comprised of three sections. Section A concerned the status
on the consumption of the herbs and herbal based products and Section B was
about the perception and knowledge on herbs and herbal based products. The
background information of the respondent was gathered in Section C.
The objective of this study was to assess the pattern of consumer behavior on
herbs and herbal based products. The Theory of Planned Behavior was used as
a guideline to develop the question in the Section B. In the questionnaire, there
were twelve criteria with five likert scale used to measure the consumer’s
preference in selecting herbal based products.
The information gathered from the questionnaire was analyzed using reliability
test, correlation test and descriptive statistic.
The status on the use of herbs and herbal based products was shown in Table
1. A total of 3270 (73.5%) out of 4452 respondents used herbs and herbal
based products in 2013. Further discussion is to focus on the 3270
respondents.
Table 1. Status on the use of herbs and herbal based products
Status
No. of respondents (%)
Never use
Used in 2013
Used in 2008 – 2012
Total
1116 (25)
3270 (73.5)
66 (1.5)
4452(100)
To test the reliability of the twelve criteria used to understand the
consumer’s preference, the Cronbach’s Alpha (α) was used. Based on George &
332
Mallery (2003) the twelve criteria were acceptable to be used in measuring the
consumer’s preference in selecting herbal based products as the α = 0.7.
Consumer preference based on twelve criteria in selecting herbal
products was shown in Table 2. There were five criteria in selecting herbal
based product that concerned the most namely easy availability (48.8%),
efficacy of the product (63.3%), previous satisfaction (70.7%), manufacturer
with GMP compliance (72.2%) and MoH registered product (76.8%).
Table 2. Respondents’ views on importance criteria in selecting herbal based product
Statement
Importance of brand
Importance of GMP
compliance
manufacturer
Importance of MoH
registered product
Importance of products'
testimonial
Importance of
manufacturer with good
reputation
Importance of efficacy
Importance of
recommendation from
family and friends
Importance of high mass
media coverage
Importance of low price
Importance of packages'
design
Importance of easy
availability
Importance of previous
satisfaction
Likert Scale Score* (Percentage)
Mode
1
3.2
2
18.9
3
20.4
4
34.9
5
22.6
0.4
2.2
3.0
22.2
72.2
5.0
0.3
2.0
1.8
19.1
76.8
5.0
2.8
18.9
20.9
36.8
20.5
4.0
1.4
11.9
14.3
47.3
25.1
4.0
0.5
1.7
4.4
30.0
63.3
5.0
3.1
12.5
27.6
36.9
19.9
4.0
4.2
20.2
28.1
31.3
16.2
4.0
2.2
15.6
32.0
27.4
22.8
3.0
2.6
15.6
22.3
35.0
24.5
4.0
0.3
3.1
7.5
40.3
48.8
5.0
0.2
0.6
2.1
26.4
70.7
5.0
4.0
* 1= very unimportant, 5= very important
From Table 3, 406 consumers with limited knowledge on herbs
mentioned that manufacturers with GMP compliance was a very important
criterion. While 1,280 and 454 consumers, with moderate and high knowledge
on herbs respectively also agreed with the same statement. There was
significant correlation between consumer preferences on importance of GMP
compliance products with their knowledge on herbs had a weak correlation.
333
When the level of knowledge on herbal products was high, there was more
tendency of respondents to choose products with GMP compliance as the most
important criterion.
Table 3. Correlation between consumer preferences on importance of GMP compliance
products with their knowledge on herbs
Level of
GMP compliance manufacturer
Total
knowledge on
Very
Very
Unimportant Moderate Important
herbs
Unimportant
Important
Very low
0 (0.00)
3 (0.09)
9 (0.28)
22 (0.67) 101 (3.09) 135 (4.13)
567
Low
3 (0.09)
22 (0.67)
19 (0.58) 117 (3.58) 406 (12.42)
(17.34)
461
1280
1827
Moderate
4 (0.12)
32 (0.98)
50 (1.53)
(14.10)
(39.14)
(55.87)
595
High
4 (0.12)
13 (0.40)
13 (0.40) 111 (3.39) 454 (13.88)
(18.20)
Very high
1 (0.03)
2 (0.06)
6 (0.18)
16 (0.49) 121 (3.70) 146 (4.46)
727
2362
3270
Total
12 (0.37)
72 (2.20)
97 (2.97)
(22.23)
(72.23)
(100)
Note: R= 0.043, p= 0.013
Source: Survey, 2013
According to Table 4, there was no significant difference between
yearly expenditure and consumer preferences on the importance of GMP
compliance. Table 5 showed that there was significant difference between
consumers’ preference on the price of herbal product in the aspect of their
yearly expenditure on the herbal based products. Consumers that chose low
price as the most important criterion had the average spending of RM442
compared to consumers with the average spending RM764 that claimed low
price was the less important criterion.
Table 4. ANOVA test for consumer preferences based on importance of GMP
compliance products from the aspect of yearly expenditure.
Likert scale
score
Very
Unimportant
Unimportant
Moderate
Important
Very
Important
Total
N
Mean
Std.
Deviation
Std.
Error
95% Confidence
Interval for Maen
Lower
Upper
Bound
Bound
Min
Max
12
484.28
799.48
230.78
-23.69
992.24
0.00
2268
72
97
727
560.15
384.34
485.99
805.80
568.71
988.52
94.96
57.74
36.66
370.80
269.72
414.02
749.51
498.97
557.97
0.00
0.00
0.00
4105
4296
11760
2362
512.15
799.00
16.44
479.91
544.39
0.00
12000
3270
503.50
839.32
14.68
474.72
532.28
0.00
12000
Note: F= 0.714, p= 0.582
334
Table 5. ANOVA test for consumer preferences based on importance of herbal based
products with low price from the aspect of yearly expenditure.
Likert scale
score
Very
Unimportant
Unimportant
Moderate
Important
Very
Important
Total
N
Mean
Std.
Deviation
73.00
763.83
1655.79
510.00
1048.00
895.00
594.35
507.92
475.99
744.00
3270.00
Std.
Error
95% Confidence
Interval for Mean
Lower
Upper
Bound
Bound
Min
Max
193.80
377.51
1150.16
0.00
11760.00
1069.59
796.59
834.97
47.36
24.61
27.91
501.30
459.64
421.21
687.40
556.20
530.77
0.00
0.00
0.00
8492.00
11544.00
12000.00
442.55
547.74
20.08
403.13
481.97
0.00
6000.00
503.50
839.32
14.68
474.72
532.28
0.00
12000.00
Note: F= 4.497, p= 0.001
CONCLUSION
The level of knowledge on herbal based products correlated with the
consumers’ concern on its safety. However, this did not affect their tendency
to spend on the herbal based products.
REFFERENCES
Rohana, A.R., Intan Nurulhani, B., Mohd Asraf, I. & Lim, H.F. 2014. Penggunaan
Herba di Semenanjung Malaysia: Tradisi atau Kontemporari?. Pp. 32-35
in M. Mastura, B.J. Chee, A.L. Tan, M.A. Nor Azah, A.R. Rohana & M.
Noorhazmira (Eds.). Prosiding Seminar Pemuliharaan & Pemerkasaan
Pengetahuan Tradisi, 23 to 24 May 2014 at Putra World Trade Center,
Kuala Lumpur.
Rohana, A.R., Lim, H.F. & Ismariah, A. (2013). Socio-economic Backgrounds and
Income Generation of Herbal Cultivators in Peninsular Malaysia. Paper
Presented at the Conference on Forestry and Forestry Products 2013
(CFFPR 2013), 11─12 November 2013, Sunway Putra Hotel, Kuala
Lumpur.
335
CONSUMERS’ PERCEPTION TOWARDS LOCAL HERBAL SUPPLEMENT
PRODUCTS
NH Kamarulzaman123 & SM Talib1
1
Department of Agribusiness and Information Systems, Faculty of Agriculture
2
Halal Products Research Institute, 3Institute of Tropical Forestry and Forest
Products, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor
Tel: 03-89474907
Fax: 03-89408213
ABSTRACT
Tongkat ali (Eurycoma longifolia), kacip fatimah (Labisia pumila), dukung anak
(Phyllanthus niruri), misai kucing (Orthosiphon stamineus), and hempedu bumi
(Andrographis paniculata) are some of the well-known local herbal food
supplements. However, one of the biggest challenges for local herbal products
is the lack of consumers’ interest in purchasing local herbs as their
supplements. Although several wellness lifestyle campaigns emphasized on the
use of local herbs in daily diet or health supplement, consumers’ awareness on
its benefits were found still lacking. Thus, the purpose of the study was to
determine consumers’ perception towards local herbal supplements. A face-toface interview with 500 respondents in Klang Valley was carried out using a
structured questionnaire. The data collected was analyzed using descriptive
analysis, chi-square analysis, and factor analysis methods. The results indicated
that majority of the respondents consumed local herbal products as
supplement. Local herbs namely pegaga, tongkat ali, and kacip fatimah were
the most preferred herbs consumed by majority of the respondents. Potential,
awareness of negative effect, effectiveness, and suitability were the factors
revealed to influence consumers’ perception towards local herbal supplement
products. This study concluded that if consumers have a well-informed
information on the potential and effectiveness of local herbal supplement
products towards their health, it will improve their acceptance of herbal
products.
Keywords: local herbs, consumers, perception, supplements
INTRODUCTION
The use of herbal medicine has been growing steadily worldwide. Considering
that Malaysia is blessed with abundant flora including medicinal plants, there is
immense opportunity locally and internationally for the Malaysian herbal
336
industry. Some of the popular local herbal plants are kacip fatimah (Labisia
pumila), pegaga (Centella asiatica), peria (Momordica charantia), misai kucing
(Orthosiphon stamineus), mas cotek (Ficus deltiodea), and limau purut (Citrus
hystrix). The use of herbs as medicine has been growing steadily worldwide.
There is a great potential for the local herbal industry to expand, as the
popularity of Malaysian herbal products is growing locally and internationally.
Under the herb Entry Point Project 1 (EPP1), the herbal industry is expected to
produce high-value products and generates income totaling RM2.2 billion of
the Gross National Income (GNI), focused on five herbs i.e. as tongkat Ali, kacip
fatimah, misai kucing, hempedu bumi, and dukung anak. Furthermore, the
trade value of the herbal industry was expected to soar over RM2 trillion by
2020. However, the herbal industry in Malaysia is still lagging behind other
countries such as China and other Association of South-East Nations (ASEAN)
countries (Ahmad & Othman, 2013). Most of the raw materials used locally
were imported from China, Indonesia, and Thailand. The local herbal industries
do not have the full support of the corporate sector, to support its
development and growth for mass production. Due to this, Malaysia is far
behind on technological advancement in herbal medicine. One of the biggest
challenges for the local herbal industry is the lack of consumer interest in
purchasing local herbs as a supplement. The consumers’ interest in herbal
products is prompted by their health problems. Consumers’ perception and
purchasing pattern are important for the development and commercialization
of herbal products. However, the true potential of local herbs is yet to be
explored. Thus, the objective of this study is to determine consumers’
perception towards local herbal supplements.
Data were collected by face-to-face interviews in Klang Valley. Sampling was
done using the random sampling method and 500 respondents were selected
for the study. The questionnaire for the study consisted of structured questions
and divided into three sections. The first section consisted of the respondent’s
profile. The second section was related to consumers’ perception towards
herbal products, and the third section covered questions related to the
respondent’s purchasing behavior of herbal products. Most of the questions
were based on a 5-point Likert scale to measure different factors that might
influenced the respondent’s overall perception towards herbal products. The
data collected were analyzed using descriptive analysis, chi-square analysis,
and factor analysis methods.
337
Socio-demographic Profiles
Socio-demographic characteristics of consumers such as gender, age, marital
status, race, number of household, income level, occupation, and level of
education were presented in Table 1. A total of 500 respondents took part in
this study. The sample group was evenly represented by males and females.
The majority of the respondents were above 45 years age group (30%), had
completed their SPM level (37%), married (56.0%), Malay (94%), and earned
below RM1,000 per month (29%). In terms of household size, 56% of the
respondents lived with two to four people in a house. In addition, 37% of the
respondents worked in the government sector.
Relationship between Respondents’ Socio-Demographic Profile and
Consumption of Herbal Products
The measurement of association between the socio-demographic profile and
consumption of herbal products were summarized in Table 2. The results
indicated that gender, age, occupation, and marital status had significant
associative relationship with consumption of herbal products. Conversely, it
was found that other socio-demographic profiles such as race, education level,
income level and household size had no significant associative relationship with
the consumption of herbal products.
338
Table 1. Socio-demographic profile of respondents
Variables
Frequency (n)
Gender
Male
250
Female
250
Age (year)
18-25
135
26-35
125
36-45
90
>45
150
Education Level
Never been to school
5
Primary
20
SRP/PMR
65
SPM
185
STPM/Matriculation
50
University
145
Others
30
Marital Status
Married
280
Single
185
Divorced
35
Race
Malay
470
Chinese
15
Indian
10
Others
5
Income level
< 1000
145
1001- 2000
130
2001-3000
90
3001-4000
30
>4000
105
Household Size
Staying alone
25
2-4
280
>5
195
Occupation
Government sector
185
Private sector
100
Self-employed
55
Housewife
45
Student
90
Unemployed
15
Others
10
Note: n=500 respondents
339
Percentage (%)
50.0
50.0
27.0
25.0
18.0
30.0
1.0
4.0
13.0
37.0
10.0
29.0
6.0
56.0
37.0
7.0
94.0
3.0
2.0
1.0
29.0
26.0
18.0
6.0
21.0
5.0
56.0
39.0
37.0
20.0
11.0
9.0
18.0
3.0
2.0
Table 2: Relationship between Respondents’ Socio-demographic Profiles and
Consumption of Herbs Product
Chi-Square
df
Significant
Decision
Test
Gender
3.793
1
0.05**
Reject H0
Age
10.646
3
0.014**
Reject H0
Race
1.162
3
0.762
Failed to Reject H0
Education Level
3.598
6
0.731
Failed to Reject H0
Occupation
13.466
6
0.036**
Reject H0
Income Level
7.969
4
0.093
Failed to Reject H0
Marital Status
7.272
3
0.026**
Reject H0
Household Size
2.424
3
0.298
Failed to Reject H0
Note: *Statistically significant at 0.01, **Statistically significant at 0.05,
***Statistically significant at 0.10
Variable
Factors Influencing Consumers’ Perception towards Purchasing Local
Herbal Products
Factor analysis was carried out to investigate various factors that influenced
consumers on purchasing local herbal products. Kaiser-Meyer-Olkin Measure
of Sampling Adequancy (KMO) and Barlett’s test of Sphericity were used to
measure sampling adequacy. Results indicated 0.719 and p=0.0000
respectively, and thus factor analysis is appropriate in this study. Based on
Table 3, five factors extracted from factor analysis. The five factors were
‘potential of herbal supplement’, ‘awareness of the negative side of herbal
supplement’, ‘consumer knowledge on local herbal supplement’, ‘effectiveness
of herbal supplement’ and ‘suitability of herbal supplement’. The first factors
indicated 22.451% of variance, the second factor was accounted for 12.586%,
third factor was 8.396%, forth factor accounted for 7.496% and the fifth factor
was 6.339%.
340
Table 3. Factor analysis result
Subvariables
Factors
Factor 1 – Potential of herbal supplement
i. Herbs is effective as nutritional supplement for
body
ii. You know about the herbs as supplement
iii. You believe that herbs is effective as
supplement
iv. The more often you consume herbal
supplement, the more you are feeling well
v. Herbal supplements are readily available in the
market
vi. Herbal supplements are sold at a reasonable
price
vii. Herbal supplements sold in stores or drug
stores is more reliable than direct sales
viii. More information about supplements from the
mass media
ix. Herbal supplements should be more campaigns
to introduce it to consumers
Factor 2 - Awareness of the negative side of herbal
supplement
i. Herbal supplements effects to the development
of the mind
ii. Herbal supplements only for those who can
afford
Factor 3 – Consumer knowledge on local herbal
supplement
i. Each herb has different nutritional value
ii. Most people do not use herbs for lack of
knowledge about herbs
iii. Herbs is good for ill treatment
Factor 4 – Effectiveness of herbal supplement
i. Most people not consume herbs because it is
not effective
ii. Most people know the benefits of herbs, but do
not believe in the effectiveness
Factor 5 – Suitability of herbal supplement
i. Herbal supplement only suitable for old people
341
% of
variance
explained
22.451
Eigen
values
Cronbach’
s Alpha
3.817
0.815
12.586
2.140
0.764
8.396
1.427
0.701
7.496
1.274
0.621
6.339
1.078
0.578
0.542
0.643
0.145
0.589
0.618
0.741
0.522
0.693
0.470
0.611
0.736
0.433
0.673
0.732
0.726
0.755
0.886
CONCLUSION
In this study, the perception and purchasing behavior of consumers towards
local herbal supplement products were determined. Results showed majority
of respondents were well-informed and consumed local herbs as supplements.
The findings will provide useful future guidelines for marketers to promote
their products, to educate and to create awareness on the usefulness of herb
in their supplements among potential consumers.
ACKNOWLEDGEMENTS
The authors would like to express their gratitude to the Faculty of Agriculture,
Universiti Putra Malaysia for supporting the research.
REFERENCES
Ahmad, S. & Othman, N. (2013). Strategic Planning, Issues, Prospects and the
Future of the Malaysian Herbal Industry. International Journal of
Academic Research in Accounting, Finance and Management
Sciences, 3(4): 91─102.
http://frim.gov.my. Accessed on 17th September 2015.
http://mardi.gov.my. Accessed on 18th September 2015.
342
THE IMPORTANCE OF LONG-TERM RELATIONSHIP
SUSTAINABILITY OF MALAYSIAN HERBAL INDUSTRY
FOR
THE
H Nurul Syahira, MN Nolila, M Norsida & M Zainal Abidin
Department of Agribusiness and Information System, Faculty of Agriculture,
Universiti Putra Malaysia
ABSTRACT
Herbal industry is one of the most promising industries in the future. The rapid
development of the herbal industry in Malaysia is reflected by its trade. Herbal
industry is one of the EPP in NKEA and government wants to improve the
product quality and marketing efforts of the herbal industry. The government
also targeted both upstream and downstream segments of the industry to help
in developing the industry. Long-term relationship has been acknowledged as
capable in increasing the competitiveness of an industry. It has been found that
retailers or buyers with long-term relationships can achieve a competitive
advantage by receiving merchandise in short supply, information on new and
best-selling products and competitive activity, best allowable prices, and
advertising and markdown allowances. Furthermore, buyers and sellers are
interdependence in the industry thus, to increase the efficiency and probability
to success, it is necessary to identify the factors that affecting the development
of long-term relationship among supply chain players in the Malaysian herbal
industry. Based on previous studies, there are a number of variables that can
contribute to the maintenance of collaborative long-term relationship such as
commitment, satisfaction, trust, power-dependence, communication,
reputation and others. It has been identified that the relationship building
efforts are theorized to enhance their membership’s commitment to the
relationship as well as the membership’s relationship behaviors. Satisfaction
has also been acknowledged as the buyer’s cognitive state of being adequately
rewarded for the sacrifices undergone in facilitating the exchange which can
help in sustaining the industry. Satisfaction also encourages greater loyalty and
a long-term working relationship. Trust is also one of the important factors that
can increase the sustainability of the industry. Trust is the willingness to rely
upon an exchange partner in whom who has confidence and facing the risk
together. Many benefits can be obtained by buyers and sellers through
development of long term relationships. Through a long-term relationship,
supply chain players in the Malaysian herbal industry are able to gain
advantages that may be not realized under a traditional relationship.
Keywords: herbal industry, long-term relationship, sustainability
343
INTRODUCTION
Malaysia is well positioned to be a key global player in the herbal medicine
industry with its biological heritage, cultural background and trade links (Arif
2002). Malaysian market for herbal products has been estimated to be worth
RM4.55 billion with a growth rate of 15% to 20% of which 90% of the raw
material used was imported (Puteh 1999). Government made development of
the herbal industry a priority under Entry Point Project 1 (EPP1) of Agriculture,
one of 12 National Key Economic Areas (NKEA). EPP1 looks at improving the
product quality and marketing efforts of dietary and herbal supplement to tap
the global demand for high-value herbal supplement and remedies. The
importance of developing the herbal industry alongside the latest global
development where the trade value of the herb sector is expected to rise by
RM2 trillion has been recognized (Natural Resources and Environment Ministry
2015). Relationship marketing is a strategy designed to foster customer loyalty,
interaction and long-term engagement which is believed can help in ensuring
the competitiveness of Malaysian herbal industry. Relationship marketing is
design to develop strong connections with customers by providing them with
information directly suited to their needs and interests. Relationship can fail to
develop or regress depending upon the actions of either party or of competing
buyers or sellers (Ford, 1980). A key strategy to increase competitive
advantage is by establishing a long-term buyer-supplier relationship (Anderson
& Narus 1990; Badaracco 1991; Ganesan 1994; Jap & Ganesan, 2000;
Noordewier et al. 1990). Christopher (1998) and Mohanty and Gahan (2012)
stated that firms that engaged in a long-term relationship are more likely to be
successful and improve the efficiency of their supply chain. A close long-term
buyer-supplier relationship allows the partners to experience benefits that
would not be realized under the traditional adversarial relationships.
MATERIALS AND METHOD
The objective of this study is to develop the conceptual framework that can be
used to analyze the importance of long-term relationship for the sustainability
of Malaysian Herbal Industry using the following methodology framework.
1. Literature review comprised of peer-reviewed research articles in the
importance of long-term relationship from international and local
journals. The articles and journals focused on techniques, framework
adaptation, methodology, and also the results obtained from the
studies.
2. Select the most suitable theory that can be used in the study.
3. Develop the conceptual framework based on the literatures.
344
Buyers' perspectives of buyer-seller relationship development by Claycomb and
Frankwick (2010) showed that the information exchange and conflict resolution
mechanisms influence the level of relationship-specific investments and buyer
uncertainty during the development of a buyer-seller relationship. Canon et al.
(2010) stated that buyer trust, supplier performance, and the control variables
explain a relatively large portion of the variance in long-term orientation.
While, Tan (2006) also supported that all of the significant supply chain
management practices positively impact performance. In developing a
framework, lot of factor need to be considered to make sure the framework
model can be used to achieve an objective and also as a guideline for the study.
Moreover, type of analysis that will be used also can affect the framework.
Some study includes satisfaction in their framework and based on the study by
Azhar et al. (2012), on customer satisfaction with hypermarket fresh food’s
characteristics showed that the satisfaction with the price of the fresh food
products influences the long term relationship with the customers. Louis
(2010) reported that, there is indirect influence on commitment via trust and
attachment to the company. Study by Jayachandran et al. (2005) showed that
by moderating the influence of relational information processes on customer
relationship performance, technology used for CRM performs an important
and supportive role. Multiple criteria approaches and techniques are based on
the development of the factors. The process to develop a conceptual
framework for the study is a three steps process involving first recognizing a
basic theory that compatible with the study. The model will develop a flow
model framework, which will expand the entire probable variable. The second
stage is to eliminate some of variables that being developed in the first stage
model framework based on the objective. Objective will determine which
variable is needed for the study and only few variables can be used. The final
step involves a development of multiple objective frameworks. The framework
developed can show the factors influencing the development of long term
relationship between commitment, trust, cooperation, satisfaction relationship
specific investment and technology as shown in Figure 1.
345
Commitment
Trust
Cooperation
Satisfaction
Relationship
Specific
Investment
Factors influencing the
development of long term
relationship
Figure 1. Cenceptual Framework
Source: Adopted from Dwyer et al. (1987)
CONCLUSION
This paper discusses the development of framework for the importance of
long-term relationship for the sustainability of Malaysian herbal industry and
implementation of the framework. The importance of the long-term
relationship has been discussed and found that it can assist the continuity of
the relationship among the supply chain players. In this study, the factors that
were introduced including commitment, trust, cooperation, satisfaction,
relationship specific investment, and technology. The output from the study is
expected to provide framework that can be used to analyze the importance of
long-term relationship for the sustainability of Malaysian herbal industry.
REFERENCES
Anderson, E. & Weitz, B. A. (1992). The Use of Pledges To Build And Sustain
Commitment in Distribution Channels. Journal of Marketing Research:
Vol 29 (1): 18─34.
Anderson, J.C. & Narus, J. A. (1990). A Model of Distributor Firm and
Manufacturing Firm Working Relationships. Journal of Marketing: Vol
54 (1): 42─58.
Dwyer, F. R., Schurr, P. H. & Oh, S. (1987). Developing Buyer-Seller
Relationships, Journal Of Marketing: Vol. 51 (3): 11─27.
346
Technology
Ford, D. (1980), The Development Of Long-Term Orientation in Buyer-Seller
Relationships. Journal of Marketing: Vol. 58 (April): 1─19.
Kalwani, M. U. and Narayandas, N. (1995). Long-Term Manufacturer-Supplier
Relationships: Do They Pay Off for Supplier Firms? Journal of
Marketing: Vol 59 (January): 1─16.
Morgan, R. M. and Hunt, S. D. (1994). The Commitment-Trust Theory of
Relationship Marketing. Journal Of Marketing: Vol 58 (3): 20-38.
Han, S. L., Wilson, D.T. and Dant, S. P. (1993) Buyer-Seller Relationships Today,
Industrial Marketing Management. Vol. 22: 331─338.
Subramaniam, C., Chandrasekaran, M. & Govind, D. S. (2010). Analyzing The
Buyer Supplier Relationship Factors: An Integrated Modeling Approach.
International Journal of Management Science and Engineering
Management: 5(4), 292─301.
Wilson, D. T. (1995). An Integrated Model of Buyer Seller Relationships. Journal
of the Academy of Marketing Science: Vol 23 (4): 335─345.
Claycomb, C., & Frankwick, G. L. (2010). Buyers Perspectives of Buyer-Seller
Relationship Development. Industrial Marketing Management: 39(2),
252─263.
Cannon, J. P., Doney, P. M., Mullen, M. R., & Petersen, K. J. (2010). Building
Long-Term Orientation in Buyer-Supplier Relationships: The
Moderating Role of Culture. Journal of Operations Management: 28(6),
506─521.
Tan, K. C. (2002). Supply Chain Management: Practices, Concerns, and
Performance Issues. Journal of Supply Chain Management: 38(1),
42─53.
Louis, D., & Lombart, C. (2010). Impact of Brand Personality on Three Major
Relational Consequences (Trust, Attachment, And Commitment to The
Brand). Journal of Product & Brand Management: 19(2), 114─130.
Jayachandran, S., Sharma, S., Kaufman, P., & Raman, P. (2005). The Role of
Relational Information Processes and Technology Use in Customer
Relationship Management. Journal of marketing: 69(4), 177─192.
347

Prosiding Persidangan Industri Herba 2015

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