Helichrysum plicatum

Optimization of the extraction of bioactive compounds
from Helichrysum plicatum DC
and formulation of phytopoduct with biological activity
Gordana Zdunić & Katarina Šavikin
Institute for Medicinal Plants Research
“Dr Josif Pančić”, Belgrade, Serbia
Greek: “helios” – sun
“chrysos” - gold
The genus Helichrysum (Asteraceae)
comprises approximately 500-600
species. Helichrysum spp. have been
used in folk medicine for at least
2,000 years in Europe and Africa in
the treatment of various medical
conditions as:
- choleretics,
- cholagogues,
- hepatoprotectives,
- for stimulation of the secretion of
gastric juice
Some members of the genus are
known for their
anti-inflammatory, anti-allergic and
antibacterial properties
Flora Europaea:
Section Virginea (DC)
1. H. amorginum Boiss. & Orph n Boiss
2. H. sibthorpii
3. H. doerfleri Rech. fil
4. H. frigidum (Labill.) Wild.
Section Helichrysum
5. H. stoechas
6. H. rupestre (Rafin.) DC.
Flora Serbica - Section Helichrysum
7. H. heldreichii Boiss.
1. H. arenarium (L.) Moench ssp. arenarium
8. H. ambiguum (Pers)
2. H. plicatum DC. ssp. plicatum
9. H. saxatile
10. H. italicum (Roth)
11. H. orientale (L.)
12. H. plicatum DC., Prodr.
13. H. arenarium (L.) Moench.
14. H. graveolens (Bieb.)
Section Xerochlaena (DC)
15. H. foetidum (L.)
16. H. bracteatum (Vent.) Andrews
.
.
Perennial herbaceous plant of height 15–40 cm
that flowers in June-August.
Broadly distributed in Europe, western Siberia,
and central Asia.
It grows in sandy soils, more rarely in open
rocky places, and is found in dry pine forests.
COMPOUNDS
Flavonoids: in particular isosalipurposide (intensive
yellow chalcone glycoside), naringenin-5-glucosylgIucoside, helichrysin A and B (C-2-enantiomeric
narigenin-5-O-glucosides, B-salipurposide)
Phthalides: including 5-methoxy-7-hydroxyphthaIides and their monoglucoside
H. arenarium (L.) Moench
Alpha-pyrone derivatives: arenole, homoarenole
Sesquiterpene bitter principles
Volatile oil (traces)
Caffeic acid derivatives
Activity
The drug has antibacterial principles, and is mildly choleretic
and mildly spasmolytic.
Indications and usage
Approved by Commission E:
• Dyspeptic complaints
Unproven Uses: The drug is used as an adjunct in the
treatment of chronic cholecystitis and gallbladder complaints
with accompanying cramps. In folk medicine, it is used as a
diuretic and for jaundice, gout, rheumatism, kidney
complaints and dropsy.
European herb native to the Mediterranean
countries.
It grows on dry, rocky or sandy ground.
Mostly used for isolation of essential oil
(treatment of burns and wounds, healing of
scars both fresh and old, anti-inflammatory
and spasmolytic).
H. italicum (Roth) G. Don fil.
•Monoterpenes: α-pinene , β-pinene, d-limonene, g-curcumene,
•Sesquiterpene: β -carophyllene
•Alcohols: linalol, geraniol, nerol, furfurol
•Esters: neryl acetate, geranyl acetate
•Aldehydes: isovaleric
•Ketones: diones, italidone, other b-diketones
•Phenol: eugenol
Chemical Features to look for:
Diketone content: italidione
Ester content: specifically neryl acetate
Sesquiterpenes content: b-carophyllene and γ-curcumene
H. plicatum:
In Turkish folk medicine
- diuretic,
- lithagogue,
- for stomach ache
In Serbian and Macedonian folk medicine
- gastric disorders
- hepatic disorders
antidiabetic
antioxidant
antibacterial
diuretic
spasmolytic activities
H. plicatum DC
OH
OH
HO
O
OH
OH
HO
HO
O
O
OH
OH
OH
OH
O
OH
apigenin
O
O
kaempferol
quercetin
These activities are mainly attributed to the presence of phenolic
compounds (phenolic acids, flavonoid aglycons and glycosides)
OH
OH
OH
HO
O
OH
O
luteolin
HO
O
OH
O
naringenin
H. plicatum extracts have been produced using different extraction conditions,
such as degree of communution, ethanol (EtOH) concentration used for
extraction (40, 50 and 60%, v/v) and solvent system used for re-extraction
EtOAc-EtOH (5:5, 9:1 and 100:0)
Extraction conditions, content of total phenolics and yield of different extracts of H. plicatum.
Degree of
Extraction
communution
solvent
(μm)
(% EtOH)
1
710
2
Re-extraction solvent
Total phenolics
Extraction
(EtOAc : EtOH)
(mgGAE/g)
yield (%)
40
9:1
264.6 ± 9.1
4.9
in toto
40
9:1
252.3 ± 8.9
2.5
3
710
60
9:1
266.9 ± 7.2
8.0
4
in toto
60
5:5
102.4 ± 6.1
10.6
5
710
50
5:5
121.6 ± 3.8
11.3
6
in toto
50
5:5
156.1 ± 5.6
9.4
7
2000
60
5:5
163.1 ± 2.6
11.7
8
2000
40
5:5
133.1 ± 2.5
8.3
9
2000
50
5:5
89.6 ± 1.4
13.3
10
710
50
9:1
280.4 ± 8.8
6.2
11
2000
50
9:1
290.8 ± 9.2
6.2
12
710
50
100:0
359.8 ± 3.5
7.0
13
2000
50
100:0
390.1 ± 9.3
6.5
Sample
Content of flavone aglycones (mg/g DW) in selected extracts of H. plicatum
Sample
Naringenin
Apigenin
Kaempferol
6
10.1
4.1
0.7
11
15.6
15.3
2.1
12
17.4
16.2
3.8
mAU
700
5
600
500
10
400
7
300
3
200
2
1
100
8
6
4
9 11
0
5
10
15
20
25
30
35
min
1, chlorogenic acid; 2, naringenin glycoside; 3, naringenin glycoside; 4, quercetin-3-O-glucoside; 5,
kaempferol-3-O-glucoside; 6, apigenin-7-O-glucoside; 7, apigenin glycoside; 8, chalcone derivative; 9,
naringenin; 10, apigenin; 11, kaempferol.
The free radical scavenging activity of selected samples on the
stable 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical.
DPPH radical scavenging activity of different H. plicatum flower extracts (samples 1-13), with Trolox as a positive control (sample 14)
The highest radical scavenging activity was observed in sample 12 with an IC50 value of 37.2 μg/mL,
while the lowest activity was shown by sample 8 with an IC50 value of 88.6 μg/mL.
Based on the total phenolic content and antiradical activity, samples 6, 11
and 12 were selected for investigation of cytotoxic activity on three
human cancer cell lines (PC3, HeLa and K562).
In vitro cytotoxic activity (IC50) of selected H. plicatum extracts (μg/mL) and flavone aglycones (μM)
Sample
HeLa
PC3
K562
6
42.0 ± 3.8
71.5 ± 3.2
72.3 ± 4.2
11
41.9 ± 4.4
42.0 ± 4.6
37.3 ± 2.9
12
42.1 ± 0.05
39.2 ± 1.1
25.9 ± 1.5
Apigenin
6.6 ± 1.7
29.5 ± 4.7
10.8 ± 3.4
Naringenin
23.0 ± 3.7
23.9 ± 1.5
22.0 ± 1.5
Kaempferol
9.0 ± 0.6
18.5 ± 3.
7.1 ± 0.9
Cisplatin*
2.1 ± 0.2
4.8 ± 0.2
7.9 ± 0.2
In conclusion, the results obtained in the present study demonstrate potent
antiradical and cytotoxic properties of several H. plicatum flower extracts, thus
indicated that this plant species could serve as a good source of antioxidants
with potential beneficial effects on human health.
Relaxant Effect of the Ethanol Extract of Helichrysum plicatum on
Isolated Rat Ileum Contractions
The aim of the present study was to evaluate the possible spasmolytic activity of the ethanol
extract of H. plicatum on rat ileum, since there is no data of the physiological effects of the
extract on isolated rat intestine.
H. plicatum extract (HPE) in cumulative concentrations (0.01–1 mg/mL) induced a
concentration-dependent inhibition of the spontaneous contractions of the rat ileum
Inhibitory effect of HPE and papaverine on spontaneous and high K+-induced contractions
in isolated rat ileum.
Comparison of dose-response curves of acetylcholine in absence
and presence of the HPE and atropine in isolated rat ileum
Comparison of dose-response curves of histamine in absence
and presence of HPE in isolated rat ileum
Comparison of dose-response curves of barium ions in
absence and presence of HPE in isolated rat ileum
The results obtained from the present study indicate that the ethanol extract of
H. plicatum flowers showed a relaxant effect on isolated rat intestine.
The extract of H. plicatum can inhibit spontaneous ileum contractions and
contractions induced by acetylcholine, histamine, barium and potassium ions.
The therapeutic effectiveness in the treatment of gastrointestinal disorders and
use in traditional medicine of this plant could be due to its spasmolytic effect.
Extractum H.plicatum siccum
25 %
Capsulac 60
60 %
Amylum Maydis
14.5 %
Magnesium stearas
0.50 %
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