program

Transcription

program

Welcome
On behalf of the organising committee, we would like to welcome you
to the second COST 926 international conference entitled 'Improving
the health value of plant foods; phytochemical optimisation' here in
Egmond aan Zee.
The two-day main event will provide a combination of keynote
lectures, on different aspects of improving the health value of plant
foods, including the effects of pre- and post-harvest treatments, food
processing and the bioavailability of phytochemicals. The programme
also includes short oral and poster presentations on relevant topics
and two open debates.
The Working and Focus Group meetings on the third day are open to
interested participants. It would, however, be helpful if you could let
the registration desk know if you wish to attend one of these
meetings. A management committee meeting will also be held on
14th October.
COST 926 is a networking partnership funded by the EU through the
European Science Foundation. The aim is to stimulate research
collaboration and cooperation within the research community,
optimise expertise and resources and facilitate the more rapid
understanding of how bioactive plant compounds can benefit human
health. The project has a strong genomic emphasis with the primary
objective of ensuring maximum benefit is gained in Europe from the
application of new technologies in cellular and molecular biology in
the study of bioactive components in fruits and vegetables, in relation
to disease prevention.
We hope that you will enjoy the conference and your stay in The
Netherlands.
Jenny Gee
Chairman of COST926
Matthijs Dekker, Ruud Verkerk and Wim van Dokkum
Local organisers
Organisation
Local Organising Committee
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Dr Matthijs Dekker (Wageningen University, The Netherlands)
Dr Ruud Verkerk (Wageningen University, The Netherlands)
Prof Wim van Dokkum (TNO, The Netherlands)
Scientific Committee
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Dr Jenny Gee (Institute of Food Research, Norwich, UK)
Dr Augustin Scalbert (INRA, Clermont Ferrand, France)
Prof. Dr. Andrew Chesson (University of Aberdeen, UK)
Dr Matthijs Dekker (Wageningen University, The Netherlands)
Dr Ruud Verkerk (Wageningen University, The Netherlands)
Prof Wim van Dokkum (TNO, The Netherlands)
Organisation Support
•
2
Drs. Ben Post, BureauPost, Doorwerth, The Netherlands
Conference venues
The conference is being held in
Hotel Zuiderduin
Zeeweg 52
1931 VL Egmond aan Zee
Lectures will be held in the Beatrix hall,
Coffee breaks and poster presentations are in the nearby hall.
Conference dinner will be held at the nearby:
Grand Café Restaurant Het Lido
Westeinde 1
1931AB Egmond aan Zee
Lido
Hotel Zuiderduin
3
Programme
Wednesday 12th October
9:00 - 10:00
Registration and coffee
10:00 - 10:10
Welcome and Introduction
Dr Jenny Gee, Chairman COST 926
Session 1
‘Selection and manipulation of the growing plant’
Chairmen: Professor Andrew Chesson and
Professor Jaroslava Ovesna
Plenary lectures (Approx 45 mins + 10 minutes questions)
10:10 - 11:05
"Genetic engineering vs traditional breeding for
manufacturing plant medicines and healthier food".
Professor Vincenzo De Luca (Brock University,
Ontario, Canada)
11:05 - 12:00
‘Agronomy as a manipulative tool’
Dr Kirsten Brandt (University of Newcastle, UK;
formerly of the Danish Inst. of Agricultural Sciences)
12:00 - 13:15
Lunch break
4
Oostveen-van Casteren ...........59
Ott ............................................31
Ovesna.....................................53
Pérez-Sacristán........................33
Periago......................... 40, 48, 52
Perkins-Veazie ...................57, 58
Poutanen..................................27
1
Poutanen ................................47
Pukalskas.................................61
Pulkkinen..................................47
Romero ....................................39
Ros............................... 39, 48, 52
Rowland ...................................20
Salopuro...................................47
Sanogo.....................................22
Sanoner....................................22
Schieber ...................................31
Schijlen.....................................25
Schirrmacher............................54
Schnitzler ..................... 41, 42, 54
Schöner, J ................................26
Schöner, R ...............................26
Schwartz ............................55, 56
Schwarz, K ...............................43
Sgaragli ....................................29
Siddiq .................................57, 58
Socaciu ....................................36
Solhaug ....................................26
Sontag................................ 55, 56
Stavělíková...............................53
Stöckmann ...............................43
Struijs .......................................59
Thielen .....................................31
Törrönen...................................27
Valoti ........................................29
van Beek ..................................61
Van Camp .......................... 35, 36
van den Broeck ........................25
Van Hoed .................................35
van Tunen ................................25
Venskutonis..............................61
Verhagen..................................18
Verhe........................................36
Verhé........................................35
Verkerk............. 17, 28, 45, 51, 60
Vidal-Guevara ..........................40
Vincken ....................................59
Voragen....................................59
Wilfried .....................................42
Woitke ................................ 41, 42
Wolffram............................. 19, 30
Zhao .........................................51
65
Ammann...................................15
Andjelkovic .........................35, 36
Bacekova .................................32
Barré ........................................22
Bengtsson ................................26
Bennett.....................................29
Berge........................................26
Bernal.......................................52
Bilger ..................................26, 44
Bitsch, I ..............................31, 37
Bitsch, R.............................31, 37
Blanco ......................................33
Blázquez ..................................33
Bobis ........................................36
Bonnema............................45, 51
Borge........................................26
Bovy .........................................25
Brandt.......................................13
Buchert.....................................27
Bunea.......................................36
Butelli .......................................38
Carle.........................................31
Carlé.........................................37
Cassidy ....................................23
Cermak.....................................30
Che...........................................46
Chen.........................................46
Collins ......................................57
Dapkevicius..............................61
De Luca....................................12
de Vos ......................................25
Dekker.......................... 28, 45, 60
Dietrich ...............................31, 49
Dolan........................................57
Dragoni.....................................29
Frank ........................................31
Frontela ....................................39
García-Alonso ....................40, 48
Gee ..........................................29
Granado ...................................33
Graßmann .................... 41, 42, 54
Gruppen ...................................59
Haffner .....................................26
Hagen.......................................26
Harbaum ..................................43
Haro .........................................39
Harte .................................. 57, 58
He…. ............................ 45, 46, 51
Herrero .....................................33
Hoffmann-Wülfing ....................44
Hubbermann ............................43
Hudecova .................................32
Jonker ......................................25
Kadrabova................................32
Kajaba ......................................32
Kallio ........................................47
Kammerer ................................31
Kaufmann.................................48
Khan................................... 57, 58
Kolehmainen ............................47
Koponen...................................27
Králová .....................................53
Krauß ................................. 41, 42
Kreck ........................................49
Kroon........................................21
Kroyer.......................................50
Kučera......................................53
Kürbel.......................................49
Leišová.....................................53
Lesser ......................................30
Li ....... ......................................46
Lombardo .................................26
López-Iniesta............................52
Lou ..................................... 45, 51
Ludwig......................................49
Madaric ....................................32
Martin .......................................38
Martínez ............................. 39, 52
Masson.....................................22
Miliauskas ................................61
Mislanova .................................32
Molthoff ....................................25
Mustranta .................................27
Nagyova ...................................32
Neacşu .....................................36
Nemeth.....................................32
Netzel, G ..................................37
,
Netzel, G .................................31
Netzel, M ............................ 31, 37
Nicolle ......................................22
Olivares ....................................52
Olmedilla ..................................33
Short communications (15 minutes plus 5 minutes discussion)
Chairmen: Dr Matthijs Dekker and Dr Fulgencio Saura Calixto
13:15 - 13:35
‘Production of health related flavonoids in tomato
fruit’
Elio Schijlen, Ric de Vos, Harry Jonker, Hetty van
den Broeck, Jos Molthoff, Arjen van Tunen & Arnaud
Bovy.
Session 2
‘Pre- and post harvest treatments and food processing’
Short communications (15 minutes plus 5 minutes discussion)
Chairmen: Dr Matthijs Dekker and Dr Fulgencio Saura Calixto
13:35 - 13:55
‘Postharvest radiation treatment to increase healthrelated quality of fruit and vegetables’.
Gunnar Bengtsson, Sidsel Fiskaa Hagen, Knut
Asbjørn Solhaug, Roman Schöner, Emanuele
Lombardo, Jennifer Schöner, Grethe Iren Borge,
Arvid Berge, Karin Haffner & Wolfgang Bilger.
13:55 - 14:15
‘Effect of enzymatic processing on anthocyanins in
bilberry and black currant juices’.
Jani M. Koponen, Riitta Törrönen, Annikka
Mustranta, Kaisa Poutanen & Johanna Buchert.
14:15 – 14:35 ‘Modelling the Consequences of Variability in Food
Production Chains on Human Health’.
Matthijs Dekker & Ruud Verkerk
14:35 - 15:25
Tea break and poster session
5
Session 3a
Open debate ‘GM technologies – the viable option?’
Author Index
15:25 – 16:40 Introduced and chaired by: Professor Klaus Ammann
(University of Bern, Switzerland) and Dr Ruud
Verkerk (University of Wageningen, The
Netherlands).
Commences with short presentations by the two
chairmen raising the issue of genetic manipulation
versus other artificial means to ‘optimise’ the intake
of bioactive plant compounds. (20-25 minute
presentations, followed by an open debate).
Session 3b
‘Pros and cons of enhancing dietary intake’
Chairmen: Dr Augustin Scalbert and Dr Ulrich Schlemmer
16:45 - 17:40
‘Potential advantages of supplementation and
supplements?’
Dr Hans Verhagen (RIVM, Bilthoven, The
Netherlands).
17:40 – 17:45
Concluding remarks
Approx 19:00
Conference dinner
Restaurant Lido (see venue information)
6
63
-3-β-galactopyranoside. Catechin and ellagic acid were found to be the most
active radical scavengers.
The activity of extracts, their fractions and purified compounds was also
tested by using various antioxidant power measurement methods, both in
model systems and edible oils. It is concluded that after some additional
testing and assessment of technological and economical feasibility of their
production some of the extracts and/or purified fractions are expected to be
promising natural products for commercial applications in health promoting
functional foods and nutraceuticals.
Thursday 13th October
8:45 - 8:50
Welcome and introduction
Session 4a
‘Bioavailability of bioactive plant compounds’
Chairmen: Dr María-Jésus Periago-Castón and Mr Mike Saltmarsh
8:50 - 9:45
‘Factors affecting bioavailabilty’
Professor Siegfried Wolffram (University of Kiel,
Germany)
9:45 - 10:40
‘Prebiotic effects of phytochemicals’
Professor Ian Rowland (University of Ulster, UK)
10:40 - 11:20
Coffee break and poster session
Short communications (15 minutes plus 5 minutes discussion).
Chairmen: Dr Mariusz Piskula and Dr Riitta Puupponen-Pimiä
62
11:20 - 11:40
‘Red wine alcohol promotes quercetin absorption and
directs its metabolism towards isorhamnetin and
tamarixetin in rat intestine in vitro’.
Stefania Dragoni, Jennifer Gee, Richard Bennett,
Massimo Valoti & Giampietro Sgaragli
11:40 - 12:00
‘Bioavailability of the flavonol quercetin is influenced
by the dietary fatty acid pattern’.
Stephanie Lesser, Rainer Cermak & Siegfried
Wolffram
12:00 - 12:20
‘Urinary excretion of antioxidants and phenolics
following fruit juice intake: influence of storing’
7
Michael Netzel, Gabriele Netzel, Dietmar-Rolf
Kammerer, Andreas Schieber, Reinhold Carle,
Undine Ott & Irmgard Bitsch.
12:20 - 13:30
Lunch break
Session 4b
‘Bioavailability of bioactive plant compounds’
Chairmen: Dr Siegfried Knassmüller and Dr Pia Knuthsen
Plenary lecture (Approx 45 mins + 10 minutes questions)
13:30 - 14:25
‘Do polymorphisms affect bioavailability?’
Dr Paul Kroon, IFR, Norwich, UK
Short communications (15 minutes plus 5 minutes discussion).
14:25 - 14:45
‘Health aspects of supplementation with a functional
food based on cereals and onion’.
Kitti Nemeth, Jana Kadrabova, Alexander Madaric,
Igo Kajaba, Anna Nagyova, Csilla Mislanova, Zuzana
Hudecova & Martina Bacekova.
14:45 - 15:05
‘Bioaccesibility of carotenoids and tocopherols from
vegetables: Effect of minimal processing (modified
atmospheres)’.
B Olmedilla, F Granado, C Herrero, B PérezSacristán, I Blanco, & S Blázquez.
15:05 - 15:35
Tea break
8
Identification of Antioxidant Components in Plants and
Assessment of their Radical Scavenging Capacity by the
Hyphenated On-Line Methods
1
2
1,2
1,2
P.R. Venskutonis , T.A. van Beek , A. Pukalskas , G. Miliauskas and A.
1,2 1
Dapkevicius ; Department of Food Technology, Kaunas University of
2
Technology, Radvilenu pl. 19. Kaunas, Lithuania; Laboratory of Organic
Chemistry, Phytochemical Section, Wageningen University, Dreijenplein 8,
6703 HB Wageningen, The Netherlands
Natural radical scavengers have attracted an increasing attention in the
scientific society during last two decades. A great number of natural
compounds present in aromatic and medicinal plants have been isolated,
purified and identified. This study summarizes the results of the search for
new antioxidants in some plants growing in the Northern Europe. Along with
well known and widely used thyme (Thymus vulgaris), less investigated
plants, such as sweet grass (Hierochloё odorata), geranium (Geranium
macrorrhizum) and bush cinquefoil (Potentilla fruticosa) were in the focus of
the present study.
Extraction and fractionation followed by structure elucidation confirmed the
presence of earlier reported and new antioxidant compounds. For instance,
in T. vulgaris together with 7 previously identified compounds two other
known radical scavenging compounds taxifolin and luteolin 7-glucuronide
were identified for the first time in this herb. A new phenylpropanoid trimer 3'O-(8''-Z-caffeoyl)-rosmarinic acid was isolated from the aqueous layer, which
remained after the hexane/water partitioning of a crude methanolic extract.
This compound demonstrated higher DPPH• scavenging ctivity than
rosmarinic acid.
Extracts from sweet grass were powerful DPPH• and ABTS•+ free radical
scavengers. Using HPLC with on-line radical scavenging detection the active
compounds, were detected in extract fractions. After multi-step fractionation
of the extract two new natural products possessing extremely strong radical
scavenging activity were isolated and their structures elucidated by NMR and
MS techniques. They were identified as 5,8-dihydroxy benzopyranone, and
5-hydroxy-8-O-β-glucopyranosyl benzopyranone.
Seven compounds were isolated and identified in the various fractions of
geranium: gallic acid, ellagic acid, 4-galloyl quinic acid, the flavonoid
quercetin and three of its glycosides, quercetin-3-β-glucopyranoside,
quercetin-3-β-galactopyranoside
and
quercetin-4’-β-glucopyranoside.
Quercetin-3-glucopyranoside and quercetin-3-galactopyranoside were the
most powerful radical scavengers.
The activity of different fractions from cinquefoil was monitored by off-line
and on-line RP-HPLC-DPPH• and ABTS•+ scavenging methods. Twelve
compounds were isolated and identified, namely ellagic acid, catechin,
quercetin-3-β-glucopyranoside, quercetin-3-β-galactopyranoside, quercetin3-β-rutinoside, quercetin-3-β-glucuronopyranoside, quercetin-3-α-arabinofuranoside,
kaempferol-3-β-rutinoside,
kaempferol-3-O-β-(6''-O-(E)-pcoumaro-yl) glucopyranoside, rhamnetin-3-β-glucopyranoside and rhamnetin
61
Thermal Degradation of Glucosinolates in Brassica vegetables
Ruud Verkerk and Matthijs Dekker; Wageningen University, Product Design
and Quality Management Group, The Netherlands; [email protected]
Glucosinolates are a group of plant secondary metabolites found exclusively
in dicotyledonous plants. The highest concentrations are found in the
Brassicaceae family comprising many commonly consumed vegetables,
condiments, forages and oil containing plants, such as cabbage, broccoli,
cauliflower, Brussels sprouts and rape. Glucosinolates and their breakdown
products are of particular interest in food research because of their alleged
anticarcinogenic properties. There are clear indications that they block
tumour initiation by modulating the activities of Phase I and Phase II
biotransformation enzymes and suppress tumours by apoptosis.
Many steps in the food production chain, such as cultivation, storage,
processing and preparation of vegetables, may have an impact on levels and
thus intake of phytochemicals such as glucosinolates (Dekker & Verkerk,
2003). Brassica vegetables are, prior to consumption, subjected to different
ways of processing, domestic as well as industrial. Domestic treatments of
the Brassica vegetables such as chopping, cooking, steaming and
microwaving have been shown to affect the glucosinolate content
considerably (Verkerk, Dekker, & Jongen, 2001; Verkerk & Dekker, 2004),
while the effects of industrial processes as freezing, fermenting and canning
are less studied.
During thermal processing of Brassica vegetables, glucosinolate levels can
be reduced because of several mechanisms: enzymatic breakdown, thermal
breakdown and leaching into the heating medium. In most studies on the
effect of thermal processing these mechanisms are not investigated
separately.
The aim of this study was to investigate the thermal degradation of individual
glucosinolates within the plant matrix. To study the degradation kinetics,
Brassica samples (broccoli, red cabbage and Brussels sprouts) underwent
different temperature treatments for various times. With the results of these
experiments, kinetic parameters have been estimated for the degradation. In
order to study only the thermal degradation, myrosinase was inactivated to
rule out the influence of enzymatic breakdown. Based on the proposed
degradation kinetics and the estimated parameters, the degree of thermal
degradation of all individual glucosinolates by standard heating conditions
(blanching, cooking, canning) was simulated.
Session 5
Open debate: ‘How do you market phytochemically rich
products? – public health needs vs. consumer desires
15:35 – 16.50 Introduced and chaired by Dr Philippe Sanoner,
(Vale de Vire Bioactives, Normandy, France) and
Professor Aedin Cassidy (University of East Anglia,
Norwich, UK).
Commences with short presentations by the
chairmen on food industry and public health
perspectives. (20-25 minute presentations, followed
by an open debate).
16:50 - 17:00
Concluding remarks and departure
M. Dekker and R. Verkerk (2003) Dealing with Variability in Food Production Chains: A
Tool to Enhance the Sensitivity of Epidemiological Studies on Phytochemicals,
Eur. J. Nutri.,42, 67-72.
R. Verkerk and M. Dekker (2004) Glucosinolates and myrosinase activity in red
cabbage (Brassica oleracea L. var. Capitata f. rubra DC.) after various microwave
treatments. Journal of Agricultural and Food Chemistry, 52, 7318-7323.
R. Verkerk, M. Dekker & W.M.F. Jongen (2001) Post-harvest increase of indolyl
glucosinolates in response to chopping and storage of Brassica vegetables.
Journal of the science of Food and Agriculture, 81, 953-958.
60
9
Friday 14th October
Working Groups, Focus Group and Management
Committee Meetings
08:45 –08:55
Overview of the day
08:55 –10:25
Working Group/Focus Group meetings
10:25 - 10:45
Tea /Coffee
10:45 - 12:00
12:00 - 13:00
Lunch
13:00 - 15:00
Management Committee Meeting
15:00 – 15:20
Tea /Coffee
15:20 – 17:00
(Continued, as necessary)
Flavonoids as an integral, minor constituent of the lignan
complex from flaxseed hulls
Karin Struijs, Jean-Paul Vincken, Harry Gruppen, Willemiek H.M. van
Oostveen-van Casteren, Alphons G.J. Voragen; Wageningen University,
Laboratory of Food Chemistry, Bomenweg 2, 6703 HD, Wageningen, The
Netherlands, e-mail: [email protected]
Lignans are a group of plant compounds with heath-promoting effects and
are, therefore, of great interest to the food and pharmaceutical industry.
Lignans are characterized by two propylbenzene units coupled through an 88′-bridge. They are found in both aglyconic and glucosylated forms (Ford et
al., 2001).
One of the richest sources of lignans is flaxseed (Wang, 2002). Flax is the
only known plant species in which lignans are part of a polymeric structure.
Hydroxy-methyl-glutaric acid (HMGA) serves as a linker between the main
constituent, secoisolariciresinol diglucoside (SDG) (Kamal-Eldin et al., 2001).
Johnsson et al. (2002) showed that in addition to SDG, also coumaric acid
glucoside (CouAG) and ferulic acid glucoside (FeAG) were liberated from the
lignan complex after alkali treatment. The aim of this study is to generate
oligomeric fragments of the lignan complex from flaxseed hulls by partial
saponification, in order to investigate how the constituents are
interconnected.
To obtain the lignan complex, defatted flaxseed hulls were extracted with
63% aqueous ethanol. After various purification steps, the lignan complex
was (partially) saponified and the constituents were separated and purified
by reversed phase HPLC. The phenolic compounds will be identified based
on MS and NMR data.
After ful saponification of the lignan complex, SDG was found to be the most
abundant lignan, but also other phenolic compounds were present in
reasonable amounts: CouAG, FeAG and herbacetin diglucoside (HDG), the
identity of which was confirmed by mass spectroscopy and NMR. Fragments
of SDG+HMGA were found in relatively high amounts in the partially
saponified samples showing that SDG is the major lignan in the complex as
described by Kamal-Eldin et al. (2001). Fragments corresponding to
HDG+HMGA indicate that also the flavonol HDG, which was reported to be
present in flaxseed (Qiu et al., 1999), is part of the lignan complex.
In conclusion, this research showed for the first time that, besides
lignans (SDG) and other phenolic compounds (CouAG and FeAG), also
flavonoids (HDG) can be an integral part of the lignan complex from flax
seed hulls.
10
59
Impact of extraction methods on the physico-chemical and
phytochemical quality of blueberry juice.
M. Siddiq1, A.A. Khan2, J. B. Harte1, K. D. Dolan1, J. K. Collins3, and P.
3
Perkins-Veazie ; (1) Dept. of Food Science and Human Nutrition, Michigan
State University, East Lansing, MI 48824 (USA); (2) Punjab Food
Department, Lahore (Pakistan); (3) USDA-ARS, South Central Agricultural
Research Laboratory, Lane, OK 74555 (USA).
The blueberry (genus Vaccinium) is a native American species high in
phytochemicals. The U.S., with nearly 80% share, is the leading blueberry
producer in the world. Michigan is the leading state with about 40% market
share. Blueberries are rich in phenolic contents—about four times greater
than in strawberries and raspberries. High phenolic content of fruit have
been strongly correlated with high antioxidant capacity. Several researchers
have suggested the significance of fruit and vegetable phenolics as dietary
antioxidants. At present, frozen blueberries are the only major processed
form being marketed. Therefore, there is a need to develop additional
processed products from this fruit. Our objective was to study the effect of
different extraction treatments on the physico-chemical, phytochemical and
sensory quality of blueberry juice.
Blueberries were washed under spray water and then mashed in Waring
blender with 2x amount of water (w/w). Juice was expressed using a rack
and cloth press from the blended/diluted mash by one of the 4 processes:
heated macerate—50 ºC, 1 hour (P-I); P-I with added Cellubrix (P-II),
Pectinex (P-III) or Crystalzyme (P-IV).
These enzymes, which were
commercial formulations of pectinases or cellulases, were added @ 0.015%
(v/v). The raw and batch-pasteurized (82.2 ºC/180 ºF, 15 sec) juice was
analyzed for TSS, pH, titratable acidity (TA), total sugars (by HPLC), clarity,
Hunter color values, total anthocyanins, total phenolics and antioxidant
capacity. A small panel of researchers evaluated the sensory quality of
pasteurized juice.
The juice yield, on whole fruit basis, ranged from 79.4% (P-I) to 87.3% (P-III).
The method of juice extraction had little or no effect on TSS, pH , TA, Hunter
color “L” or “b” values. The method of juice extraction had significant effect
on Hunter color “a” values, which were in the range of 1.0 to 3.1. The juice
clarity, expressed as % transmittance, was highly process-dependent,
ranging from 30.5 to 53.7 in P-I and P-IV juice, respectively. The highest
total sugars, anthocyanins and phenolic contents were observed in P-I juice,
followed by that in juice from P-IV and P-II. The juice extraction method or
pasteurization had minimal effect on antioxidant capacity, determined as
ferric reducing absorbance power (FRAP), with juice from both P-I and P-II
exhibiting highest FRAP values. Juice from P-I, followed by that from P-III
received highest sensory scores for flavor and overall acceptability. The
pomace, a byproduct of juice extraction, owing to its 6-times more total
phenolics and antioxidant capacity than in juice, also has a great potential to
be processed into an ingredient for use in different foods.
58
Abstracts
Plenary Lectures
In chronological order
11
Genetic engineering vs traditional breeding for manufacturing
plant medicines and healthier food
Vincenzo De Luca, Brock University, Biology Department, 500 Glenridge
Avenue, St Catharines, Ontario, L2S 3A
OH
N
C2H5
N
H
H3CO2C
Madagascar
Periwinkle
N
H3CO
N
H3C
H
OAc
H
HO CO2CH3
Vinblastine
Antineoplastic agent
Plants make an incredible array of natural products that have an unlimited
variety of biological effects on the organisms that interact with them. Our
research has focused on the biosynthesis of various natural products such
as the highly toxic and anticancer monoterpenoid indole alkaloids of the
Madagascar periwinkle and the floral sweet aroma of methyl anthranilate in
Concord grape. The exotic periwinkle and grape chemistries appear to be
produced as a result of specific tissue-, cell-, environment- and
developmentally regulated controls that define the timing and patterns of
accumulation of each class of natural product. Our interest in characterizing
these controls have prompted the development of laser capture microdissection technology together with differential cell-extraction techniques for
understanding how such controls permit the precise accumulation of these
secondary metabolites within the developmental program of each plant
species. The results will illustrate how such technology can be applied to the
understanding of cellular and biochemical specialization that leads to the
manufacture and accumulation of natural products within multicellular
organisms.
In the broader context, this new knowledge about biochemical specialization
should be applicable to using plants for manufacturing natural products by
transferring whole biochemical pathways into specialized plant cells.
Alternatively, the biochemical and molecular information gained from
pathway characterization can be used in molecular breeding efforts to obtain
high yielding lines. Both these approaches are yielding valuable information
about the flexibility of these metabolic pathways for enhancing the production
and accumulation of desired secondary metabolites. For example it is
conceivable that pathways for making useful drugs that cure or that prevents
disease will be transferred to food crops. However, in order to make a
connection between the food we eat and its effect on human health,
significant new efforts need to be made by the medical establishment to
identify how natural products in food may prevent disease.
12
Development of lycopene-rich food products with added
watermelon juice or juice concentrate
1
1
2
1
3
M. Siddiq , J. B. Harte , A.A. Khan , K. D. Dolan , J. K. Collins , and P.
Perkins-Veazie3. (1) Dept. of Food Science and Human Nutrition, Michigan
State University, East Lansing, MI 48824 (USA), (2) Punjab Food
Department, Lahore (Pakistan), (3) USDA-ARS, South Central Agricultural
Research Laboratory, Lane, OK 74555 (USA).
Watermelon is rich in lycopene, a carotenoid reported to have a protective
effect against certain cancers and cardiovascular diseases. At present, this
fruit is primarily consumed in its whole or fresh-cut form; even its uses in
foodservice rarely go beyond these forms. There is a need to develop
processing methods for watermelon juice and concentrate that have minimal
effect on lycopene degradation and sensory attributes. The objectives of the
present study were to: (1) standardize watermelon juice and concentrate
processing method(s) that optimized lycopene, and (2) develop acceptable
quality lycopene-rich food products using watermelon juice and/or juice
concentrate as an ingredient.
Whole seedless watermelons were washed, sanitized (5-min dip in 160 ppm
chlorine), and manually peeled. The peeled fruit was mashed and juice
extracted from: cold macerate (P-I), heated macerate—50 ºC for 30 min (PII), and P-II treated with 0.01% commercial pectinase (P-III). Juice from P-I
was concentrated to: 42 ºBrix and pasteurized (85 ºC, 15 seconds)—for
blends with apple, white grape, cherry, pineapple, and mango juices; or 65
ºBrix—for use as an ingredient in yogurt, muffins, brownies, and bread.
Juice and juice concentrate was evaluated for lycopene contents, TSS, pH,
titratable acidity, clarity (juice only) and Hunter color values. The quality of
juice blends and other products was evaluated for sensory attributes of color,
aroma, flavor, and overall acceptability.
Method of juice extraction had little or no effect on pH, titratable acidity and
Hunter color “L” or “b” values. However, juice from P-I and P-III had highest
Hunter “a” values and % transmittance values, respectively. Heating of
macerate resulted in 18% lower lycopene contents in the juice.
Pasteurization of juice did not affect the lycopene content significantly. On a
scale of 1-9 (9-like extremely), juice blend containing 25% pineapple and
75% watermelon juice had the highest score of 7.25 for overall acceptability
by a 110-member consumer panel; all other juice blends scored over 5.
Yogurt, muffins, brownies, and bread made with 3% addition of 65 ºBrix
concentrate showed better acceptability than those made with 5% level, as
evaluated by a small trained panel. Lycopene was fairly stable during baking
or yogurt making process. The addition of watermelon juice concentrate
resulted in an attractive light red-color in these products. Based on these
results, it is concluded that the addition of watermelon juice or concentrate
presents an excellent opportunity to manufacture lycopene-rich juice blends
and dairy and bakery products.
57
Isoflavone Pattern in Soy- and Red Clover Based Nutritional
Supplements
Heidi Schwartz and Gerhard Sontag; Department of Analytical and Food
Chemistry, Währinger Straße 38, University of Vienna, Austria
Advertized as multifunctional natural remedies isoflavones have conquered
the market in the form of soy- and red clover based nutritional supplements.
However, although the biologically active compound is the aglucone, and
bioavailability, pharmacokinetics and metabolism depend on the isoflavone
pattern and the conjugation status of the individual isoflavones [1]
manufacturers neglect to state the content of the individual isoflavones and
whether the claimed total isoflavone contents are expressed in aglucone
equivalents or as a sum of free aglucones and glucosides.
The objective of this work was to determine the isoflavone pattern of different
soy- and red clover based nutritional supplements and to compare the
experimentally obtained contents of both total aglucones and free aglucones
plus glucosides with the manufacturer’s claim. The total aglucone content
was determined by acid hydrolysis of a 20 mg aliquot of 4-6 pooled
homogenized tablets or capsule contents in 9 ml of ethanol and 1 ml of 10 M
hydrochloric acid for 90 min followed by pH-adjustment to 3-4, stepwise
dilution to 250 or 500 ml and RP-HPLC analysis of a 10 µl aliquot in gradient
elution mode. Detection was performed coulometrically using 7 channels in
the potential range between +300 and +700 mV. The proportion of free
aglycones was determined by ultrasound assisted extraction of a 20 mg
aliquot with 70% aqueous ethanol, dilution and HPLC analysis.
Red clover based products contained mainly formononetin and biochanin A
whereas soy preparations were rich in daidzein, genistein, and, to a lesser
degree, glycitein. Soy- and red clover containing supplements showed
variable proportions of the individual isoflavones reflecting, in part, the ratio
of the original soybean- and red clover extracts used to manufacture the
product. Contrary to red clover in which isoflavones occur naturally as
malonyl- and acetyl-ß-glucosides or as ß-glucosides [2] red clover based
supplements contained almost exclusively isoflavone aglucones. Conversely,
preparations containing soy showed variable proportions of conjugated
isoflavones. The only purely soy based product contained 96% of its
isoflavones in conjugated form.
The experimentally determined isoflavone aglucone content was in
agreement with or higher than the manufacturer’s claim in 5 out of 9 cases.
Expression of the isoflavone content as a sum of free aglucones and
glucosides raises the number of matched claims to 7. As isoflavone
aglucones are the biologically active form and in order to facilitate the
comparison of different commercially available isoflavone supplements
uniform regulations about labelling are required.
K.D. Setchell, N.M. Brown, P. Desai, L. Zimmer-Nechemias, B.E. Wolfe, W.T.
Brashear, A.S. Kirschner, A. Cassidy, J.E. Heubi, J. Nutr. 131 (2001) 1362S.
Q. Wu, M. Wang, J.E. Simon, J. Chromatogr. A 1016 (2003) 195
56
Agronomy as a manipulative tool
Kirsten Brandt, University of Newcastle upon Tyne
The general ecological mechanisms that determine the interactions between
availability of resources (nutrients, water) and synthesis of defence related
secondary metabolites in the plants will be presented. In short, under normal
conditions in nature, where growth is limited by a relatively constant
availability of nitrogen at a moderate level, plants strike a balance between
defence and growth. They use a fraction of resources such as carbohydrate,
N and S to reach a genetically determined normal level of defence
mechanisms, which will prevent or quickly overcome most types of
infections. If the local availability of nitrogen happens to increase (e.g. from
decomposition of animal faeces), the growth rate increases, and the balance
shifts. The concentration of defence compounds in the plant decreases, and
the plant now relies more on compensatory growth and less on resistance as
response to infections. (Stamp 2003 Q. Rev. Biol. 78, 23-55).
While most of this information comes from investigations of wild plants in
nature, mainly in the context of understanding the consequences of pollution
with nutrients, the same mechanisms appear to operate in agricultural
settings. Most of these data are about antioxidants, where high
concentrations are required to induce a biological response in human cells.
Compounds such as polyacetylenes, alkaloids, furanocoumarins etc. have
higher biological impact per molecule, and are more important for the plant
defence. In cases where the bioavailability of these defence compounds is
similar to or higher than for the antioxidants, a given concentration is likely to
result in a stronger impact on human health (Brandt et al. 2004, TIFS 15,
384-393).
If a farmer wants to maximise the concentration of a bioactive compound in
the crop (or to optimise it, if too high levels are detrimental for food quality or
human health), is it therefore important to reduce the fertilisation intensity, in
particular to avoid periods with high excess of nutrients such as N. Other
ways of manipulating the balance is by partial drying and other methods that
impose controlled levels and types of stress on the plants. Existing models
relate to bioactive compounds that defend the plant against diseases and
pests, including glucosinolates and tannins. Compounds with other
physiological roles, such as sunscreens or involved in photosynthesis or
signalling (colour, scent), will act differently, so their highest levels may
correspond to a different range of resource availability than for the defence
compounds (Brandt & Mølgaard 2001, J.Sci. Food Agric. 81, 924-931). This
would be the case for most flavonols and carotenoids. Compounds that store
minerals, such as phytate, will increase with the input of the mineral up to
quite high levels,
This implies that existing defined types of agronomic practice, such as those
used in organic farming, will have consistent and predictable consequences
for concentrations of bioactive compounds in the crop (when the variation
due to genotype and climate is taken into consideration). Even though
neither system requires the use of a particular amount or timing of nutrients,
experienced farmers cultivating plants with or without pesticides will learn
how much fertiliser to use and how for their system, creating consistently
higher levels of defence compounds in organic fresh plant foods (typically
10-50% more than corresponding conventional) (B & M 2001).
13
To predict the effect of novel practices or new technologies, the most
important consideration is therefore how they will affect the resources
available to the plant. Based on epidemiological data, the increase in life
expectancy by a doubling of the vegetable intake has been estimated to 1-2
years (van’t Veer et al. 2000 Pub. Health Nutr. 3, 103–107). So if the
bioactive compounds are responsible for this effect, an increase of 10-50%
will increase the life span by 1-12 months. The decrease in yield and thus
increase in the cost of raw material is around 30%. If multiplied with the
number of people potentially affected, this benefit/cost ratio is much better
than for some if the existing food safety measures, e.g. against BSE and
trichinosis.
Determination of the Isoflavone Content of Soy- and Red Clover
Based Nutritional Supplements by High Performance Liquid
Chromatography Coupled with Coulometric Electrode Array
Detection
H. Schwartz, G. Sontag*; Department of Analytical and Food Chemistry,
Faculty of Chemistry, University of Vienna, Waehringer Strasse 38, A- 10 90
Vienna, Austria; [email protected]
Promoted as alternative to traditional hormone replacement therapy
isoflavone nutritional supplements have conquered the market. However,
there is little regulation about labelling and product data neither state the
content of individual isoflavones nor do they allow conclusions whether the
manufacturer’s claim is expressed in aglycone equivalents or as a sum of
free aglycones and glycosides. In the present study 11 soy- and / or red
clover based supplements were analyzed for their content of total isoflavone
aglycones and free aglycones plus glycosides using high performance liquid
chromatography in gradient elution mode coupled with coulometric electrode
array detection after acid hydrolysis or direct extraction with 70% aqueous
ethanol. The ratio of the individual isoflavones and the conjugation pattern
turned out to be highly variable in the investigated products. Whereas red
clover based supplements contained isoflavones exclusively in free form soy
containing preparations showed variable proportions of conjugated
isoflavones. The experimentally determined isoflavone aglycone content was
in agreement with or higher than the manufacturer’s claim in 6 of the 11
investigated products. Expression of the isoflavone content as a sum of free
aglycones and glycosides raised the number of matched claims to 8.
Following the intake recommendations given by the manufacturers between
12.0 and 90.8 mg isoflavone aglycones would be consumed every day.
Keywords: Isoflavones; Soy; Red
Coulometric electrode array detection
14
clover;
Supplements;
RP-HPLC;
55
Influence of UV-light on the antioxidative capacity of extracts
from Spinacia oleracea L. and Gynura bicolor (Willd.) DC.
G. Schirrmacher, W. H. Schnitzler and J. Graßmann; Technische Universität
München, Chair of Vegetable Science, Duernast II, 85350 FreisingWeihenstephan
When UV-light, especially UV-B, reaches the leaves of plants, this can lead
to an augmentation of secondary plant metabolites (SPM). These
compounds are, for example, phenols (phenolic acids or flavonoids) or
carotenoids. Since these compounds are valuable for human nutrition, we
investigated if the antioxidative capacity (AC) is altering when plants (spinach
and gynura) are grown under a controlled environment and are radiated with
UV-B. We investigated the AC of phenolic and carotenoid extracts of the
treated plants in two biochemical testsystems: Xanthin/Xanthinoxidase- and
ABTS-system. Collateral determination of Total Phenolics by means of FolinCiocalteu and HPLC-analyses helped to explain the results.
We found that the content of SPM was altered by a treatment with UV-B. The
treatment lead to an increase in total phenolics, whereas carotenoids reacted
with a slight decrease. The AC of phenolic extracts from Gynura was
increased in both in vitro testsystems. Spinach extracts only showed positive
results in the ABTS-System. The AC of carotenoid extracts of spinach was
not effected by a light-treatment. For Gynura a slight rise could be observed.
HPLC-Analyses confirmed that the compostion of phenolic SPM isn´t
changed by random, but to protect the plant from UV-B.
Biofortification of crops, the need and the questions
Klaus Ammann, University of Bern, Switzerland
The Golden Rice
It is clear that with SGR2 (second generation of Golden Rice) replacing white
rice in the diet, a very significant decrease in the morbidity and mortality from
Vitamin A Deficiency is to be expected.
Within the Golden Rice network (see Dubock 2003 for more detail) in Asia
the several Syngenta Golden Rice transformation events, and others, will be
investigated in parallel. Informed event selection will then be made, in the
light of a variety of studies being conducted in parallel with rice breeding
work. At that stage the intent remains to select the most suitable
transformation event (and destroy the rest) and use the selected event to
introgress into locally adapted rice varieties important for each area where
VAD is a problem. The purpose of this ‘single transformation event’ strategy,
developed by the Humanitarian Board, (see www.goldenrice.org), is
that regulatory costs should be saved, and speed of product delivery
increased.
The potential economic impact of the introduction of Golden Rice in Asia has
been estimated, (incidentally without any knowledge of the high expressing
Syngenta transformation events), in a World Bank Policy Research Working
Paper as in excess of $15.0 billion annually – due to raised agricultural
productivity – with a negligible impact on exports, as most Asian rice is
grown for local consumption. (Anderson, Jackson and Nielsen, 2004).
Dubock AC, 2003. Learnings From Public: Private Partnerships for GM Crops. 7th
ICABR International Conference on Public Goods and Public Policy for Agricultural
Biotechnology Ravello (Italy) June 29 to July 3, 2003. 8 pp
Potrykus, I. (2001) Golden rice and beyond. Plant Physiology, 125, 3, pp 1157-1161
Potrykus, I. (2001) The 'Golden Rice' Tale. In Vitro Cellular & Developmental BiologyPlant, 37, 2, pp 93-100
Potrykus, I. (2003) Nutritionally enhanced rice to combat malnutrition disorders of the
poor. Nutrition Reviews, 61, 6, pp S101-S104. http://www.botanischergarten.ch/
Rice/Potrykus-Nutrition-2003.pdf
Negative factors:
There appear to be three major negative influences on rapid progression of
Golden Rice to an approved product which can be used. The first of these is
an intergovernmental agreement: the Convention on Biodiversity, and
particularly its interpretation.
The second is principally a European
governmental philosophy: the ‘precautionary principle’, and, again,
particularly its interpretation. And the third is the vociferous opposition to GM
crops, of (some) NGO’s, and their impact on ‘political will’, important in
developing countries for GM crop acceptance.
An important project in Africa, which has just started,
on the biofortification of Sorghum.
Africa Harvest’s plan is to implement needs-driven programs and provide
practical solutions to challenges facing resource-poor smallholder farmers in
rural communities. The Foundation believes in undertaking projects that
54
15
focus on the Whole Value Chain (WVC) Strategy, which it has developed
and refined, over the years. The uniqueness of the WVC Strategy is that it
looks at agricultural products and projects through an inverted pyramid; this
means that an end user or customer is identified before focusing on issues
such as agricultural production (inputs such as seedlings, fertilizers and
water), product distribution and marketing. This way, project beneficiaries are
able to make money from the sales of whatever is produced in excess of
home consumption requirements.
Sorghum (Sorghum bicolor (L) Moench) originated in Africa, probably in
several centres on the continent over a period of years. Africa grows over
50% of the world’s Sorghum. The 10 biggest Sorghum producers in Africa
are: Nigeria, Sudan, Ethiopia, Burkina Faso, Niger, Tanzania, Mali, Chad,
Egypt and Uganda. These countries also contain some of the centres of
diversity for Sorghum.
It is the dietary staple for more than half billion people around the world and
is the sixth most planted crop in the world, grown on more than 100 million
acres each year worldwide and currently produces about 60 million metric
tons of grain each year;
Low in protein quality due to its low content of essential amino acids, such as
lysine. The reliance on Sorghum as an important food in regions of Africa
and Asia can result in problems associated with malnutrition, especially in
children;
Closely related to corn/maize. It is the most related of all cereals, so genetic
properties of each are very closely related;
This crop’s importance is underlined by the millions of smallholder farming
communities in developing countries of Africa and Asia. Conventional
breeding methods continue to be used to improve and increase Sorghum
productivity. In view of its importance in meeting nutritional needs of millions
of people and limitations of crop breeding to develop nutritionally fortified
varieties using only conventional means, modern genetic engineering
techniques have to be utilized to develop fortified Sorghum cultivars that are
high yielding, rich in essential amino acids and acceptable to farmers and
consumers.
All those complex questions can best be discussed within a Systems
Approach, a new way of sorting out 'wicked problems' with social
implications.
Ammann K, Papazova Ammann B. 2004. Factors Influencing Public Policy
Development in Agricultural Biotechnology. In RISK ASSESSMENT OF
TRANSGENIC CROPS., ed. S Shantaram, pp. 1552. Hoboken, NJ, USA: Wiley
and Sons. http://www.botanischergarten.ch/Wiley/Factors-Discourse-Wiley.pdf
Evaluation of garlic (Allium sativum L.) genetic resources by
AFLP and sequencing
Jaroslava Ovesna, Leona Leišová, Helena Stavělíková, Jana Králová,
Ladislav Kučera; Research Institute of Crop Production, Praha – Ruzyněm
Czech Republic, e-mail: [email protected]
Institute of Chemical Technology Prague– Dejvice, Czech Republic
Genetic resources are considered a valuable heritage of mankind. Genetic
resources are collected, maintained and evaluated for their further use for
improving the potential of crop species. Collections of genetic resources
usually contain thousands of accessions of various origin. In the Genebank
operated by Research Institute of Crop Production collections od vegetable
genetics resources are available, garlic (Allium sativum L.) is represented by
hundreds of accessions from different parts of the world.
Garlic is among the oldest known horticultural crops. In the Old World,
Egyptian and Indian cultures referred to garlic 5000 years ago. Garlic grows
wild only in Central Asia (centered in Kyrgyzstan, Tajikistan, Turkmenistan,
and Uzbekistan) today. Earlier in history garlic grew wild over a much larger
region and, in fact, wild garlic may have occurred in an area from China to
India to Egypt to the Ukraine. Garlic is used as a flavoring in cooking and
pickling, sometimes in the form of whole or grated cloves and sometimes in
the form of a cooked extract, as in sauces and dressings. In medicine, garlic
is used as a digestive stimulant, diuretic, and antispasmodic. Other possible
mild medicinal uses are under investigation
Selected accessions representing germplasm from different parts of the
world were characterized with (1) respect to the content of biologically active
compounds using HPLC, (2) overall variability DNA level using AFLP and (3)
variability of the alliinase underlying DNA sequence from genotypes differing
in the content of biologically active compounds.
Within the work, suitable set of selective primer combinations for AFLP was
chosen. Variability was sufficient to develop dendrogram showing
association among genotypes. Correlation between alliin content and DNA
variability will be discussed, as well as variability within the alliinase coding
sequnce.
Acknowledgement: The work was supported by project COST926, poejct of the Czech
Ministry of Agriculture 0002700602 and Ministry of Education and Youth and Sport
1P05OC5054
go to the website of the new project: http://www.ahbfi.org/
16
53
Potential advantages of supplementation with folic acid or folate
Olivares AB, Bernal MJ, Ros G, Martínez C, López-Iniesta MD and Periago
MJ; University of Murcia. Dpt. Of Food Technology, Food Science and
Nutrition, Spain; [email protected]
OBJECTIVE
In Spain is not obligatory to fortify food products with folic acid, but the
number of enriched foods with this vitamin is increasing every day. The aim
of the present study was to evaluate the evolution in the presence in the
market of these products, to analyze the folate form used for fortification and
to verify the folate content comparing with the content expressed in the
product label.
METHODOLOGY
•
Periodic revision of enriched products in markets and supermarkets
from Murcia as evolution index of use of folic acid (or other folate
forms) in food fortification.
•
Quantification of folate content in these food products by a validated
HPLC Method (1, 2).
•
Comparison between results from HPLC Method and label
information.
RESULTS
The present study developed between 2000 and 2005 about enriched
products in the market (dairy products, breakfasts cereals, snacks,
vegetables soups or juices) have evidenced an important enhancement in
the presence of these folate enriched products. In this way it has been
showed that mostly of these products provide between 10 to 50% per service
of the RDA for folic acid. The lowest percentages were found in infant
products while the highest were found in adult products, like enriched milk or
enriched breakfast cereals. The form detected was the synthetic form of folic
acid named pteroylpolyglutamic acid or PGA.
CONCLUSIONS
Due to the relevant importance of this vitamin and its massive presence in
the diet it is necessary evaluate the real content of folic acid in these
products, their contribution in the total folate intake, as well as it is also
necessary to revise the current Food Composition Tables (FCT) in order to
consider the addition of these products in the FCT.
New research of potential strategies to fortify with naturally occurring folate
forms (L-methylfolate) or biofortification are needed.
References:
1. Vahteristo et al., 1996. J. Agric. Food Chem. 44 (2): 477-482
2. Olivares AB et al., 2004. An. Vet. (Murcia). 20: 59-73.
Acknowledgements:
European Proyect: “Folate: From food to
functionality and optimal health” (QLK1-1999-00576). Government of Spain:
AGL2000-2482-CE and AGL2003-03598 proyects.
52
Health-protective efficacy of food derived from plants enriched
in bioactive compounds; a chain perspective
Ruud Verkerk, Wageningen University, Product Design and Quality
Management Group, The Netherlands. e-mail: [email protected]
There is a strong interest of consumers for food products with a supposed
effect on their health and well being. This has led to a growing number of
product introductions by food industry containing elevated concentrations of
certain bioactive compounds. These bioactive compounds are generally
defined as compounds in foods that deliver a health benefit beyond basic
nutrition. A process has been proposed for determining the efficacy of
consumption of a bioactive component or a food enriched in a bioactive
component for the reduction of a certain disease (Finley, 2005).
First, the compounds must be chemically identifiable and the proposed
health benefit must have measurable endpoints. Second, the efficacy of the
compound has to be established in human, in-vitro and animal studies
(mechanistic hypothesis). Third, the bioactivity and thus a protective effect of
a compound has to be investigated when consumed in the form of an actual
food product.
Usually, initial indications of health-protective efficacy of foods often are
based on epidemiologic studies. However, epidemiology suffers often from
confounding factors as uncertainty and variation, both in the product as well
as in the consumers, causing difficulties in the prediction of the alleged
protection of the actual compounds in the foods (Dekker & Verkerk, 2003).
Accurate assessment of dietary intake of bioactive compounds is of
importance to investigate associations between vegetable/fruit consumption
and derived products and disease incidence, based on a dose-response
relation of the actual compound intake versus disease risk. It is essential to
realize that the ultimate protective effect of a plant or food, enhanced in a
bioactive compound and delivered to the consumer, depends on the action of
all the actors involved in the food chain. For example, it is demonstrated that
many steps in the food production chain, like cultivation, storage, processing
and preparation of the vegetables can have large impact on the levels and
thus intake of the bioactive glucosinolates (Dekker et al., 2000). Therefore,
adequate information needs to be available to i) prove the compound is in
the food in the amount and form claimed; ii) the compound is accessible from
the food; and iii) the compound is bioavailable from the food.
An integrated approach of health/nutrition science and plant/food science is
crucial for optimizing the whole production chain in order to deliver food
products to enhance consumer health and well being in an effective and
reliable way.
J.W. Finley (2005) Proposed criteria for assessing the efficacy of cancer reduction by
plant foods enriched in carotenoids, glucosinolates, polyphenols and selenocompounds. Annals of Botany 95, 1075-1096.
M. Dekker, R. Verkerk and W.M.F. Jongen (2000) Predictive modelling of health
aspects in the food production chain; a case study on glucosinolates in cabbage.
Trends in Food Science 11(4/5), 174-181.
M. Dekker and R. Verkerk (2003) Dealing with variability in food production chains: A
tool to enhance the sensitivity of epidemiological studies on phytochemicals, Eur.
J. Nutri.42, 67-72.
17
Potential advantages of supplementation and supplements?
Exploitation of existing genetic variation in B. rapa germplasm
for glucosinolate composition using molecular genetic tools.
Dr. Hans Verhagen, Head, Centre for Nutrition and Health (PB 84), National
Institute for Public Health and the Environment (RIVM), Bilthoven, The
Netherlands. E-mail: [email protected]
Lou P, He H, Zhao J, Verkerk R and Bonnema A.B.; Laboratory of Plant
Breeding, Wageningen University; Product Design & Quality Management
Group (PDQ, Dpt Agrotechnology and Food Sciences
1.
Nutrition and health are important for consumers, scientists, authorities
and industry. Nowadays there is much interest in potential health
benefits of food ingredients, foods, food supplements and diet. The
beneficial as well as the adverse effects of all these need to be critically
assessed: toxicological risk assessment and assessment of efficacy.
Moreover, the combination assessment of beneficial and adverse effects
of one ingredient, food (supplement) or diet needs to be assessed: riskbenefit analysis.
2.
The well-established risk analysis paradigm intails three parts:
•
risk assessment (hazard identificaton & characterization, exposure
assessment, risk characterization)
•
risk management
•
risk communication
3.
Efficacy assessment can be done along a similar line. A very important
step is the establishment of scientific criteria for the substantiation of
health claims on foods: the EU-Passclaim project has published its final
consensus report in June 2005.
4.
The combined assesment of risk and benefits combines the
assessments of health risks and efficacy into one. For foods and
supplements this comprises a.o.:
•
post-launch monitoring of functional foods, supplements and
ingredients
•
fortification guidelines for vitamins and minerals, incl. max levels for
safe addition to foods and food supplements
•
a common currency (e.g. using DALY’s) to weigh risk and benefits
of functional foods, supplements and ingredients in one assessment
•
a ranking of risks and benefits of foods and ingredients, thereby
discriminating between major and minor determinants of nutrition
and health (e.g. as recently done in the report “Measuring Dutch
Diets - healthy diet and safe food in the Netherlands”)
5.
A new paradigm is applicable for risk-benefit analysis, the former of
which is the challenging realm of scientists:
•
risk-benefit assessment
•
risk-benefit management
•
risk-benefit communication
The genetic diversity of a collection of 161 Brassica rapa accessions
representing different morphotypes among the cultivated B. rapa, including
traditional and modern cultivars, and breeding materials from geographical
locations all over the world was investigated. Most interesting information
revealed by the phenetic trees is that different morphotypes are often more
related to other morphotypes from the same region (East Asia versus
Europe) than similar morphotypes from different regions, suggesting either
independent origin and or a long and separate domestication and breeding
history in both regions. F2 and Doubled Haploid populations have been
established from crosses between phylogenetic diverse accessions with
extreme phytate and mineral concentrations. DH populations are obtained
from a cross between Yellow Sarson and Pak Choi, Yellow Sarson and
Vegetable Turnip and between Yellow Sarson and Mizuna. Both the Double
Haploid (DH) populations and the collection of accessions are the basis for
the Brassica rapa research program. A genetic map of the DH populations of
AFLP and microsatellite markers will be presented.
Focus of the genetic analyses will be on the genetic dissection of health
promoting glucosinolates that will be monitored throughout the production
chain. Glucosinolates are sulfur containing plant metabolites that have anticarcinogenic properties and are largely responsible for the typical flavor and
odor of Brassica species. Glucosinolate levels in leaf tissue of parental lines
used to construct the DH populations are qualitatively and quantitatively very
different. Vegetable turnip has an extreme high concentration compared to
Pak Choi, Yellow Sarson and Mizuna. The aliphatic glucosinolates
gluconapin (NAP), gluobrassicanapin (GBN) and the aromatic gluconastutiin
(NAS) are the most dominant glucosinolates in the Brassica rapa group.
QTLs for glucosinolate composition and a number of agronomic traits will be
presented.
http://www.rivm.nl/en/aboutrivm/organization/vgc/cvg/index.jsp
18
51
Evaluation of Polyphenols and Antioxidant Activity of Grape
Seed Meal and Grape Seed Oil
Gerhard Kroyer; Division of Natural Products and Food Chemistry, Institute
of Chemical Engineering, Vienna University of Technology, Getreidemarkt 9,
A-1060 Vienna, Austria
There is a growing interest in the food industry and in preventive health care
in the development and evaluation of natural antioxidants from plant
materials. Grape seeds contain considerable amounts of polyphenol
substances which have been reported to have antioxidant and radical
scavenging activity. Furthermore, grape seeds are extracted to yield grape
seed oil which is known for its beneficial nutritional value owing to its high
content of linoleic acid.
The objective of this study was to determine the content of total polyphenols
in grape seed oil and the remaining grape seed meal as residue of the grape
seed oil extraction, and to evaluate their appropriate antioxidant and free
radical scavenging properties.
The dry grape seeds were finely crushed. Grape seed oil was extracted with
n-hexane and analysed together with the remaining defatted grape seed
meal for their content of total polyphenols according to the Folin-Ciocalteu
method. The radical scavenging capacity was evaluated by the DPPH radical
scavenging method in terms of their antiradical power ARP.
Defatted grape seed meal contains significant high amounts of polyphenols
(47,3 mg/g) which also could be detected in grape seed oil in remarkable
quantities (8,9 mg/g). Correlating with the total polyphenol content the radical
scavenging capacity of defatted grape seed meal extract was found to be
significant high (ARP: 56), whereas grape seed oil still shows effective
radical scavenging properties (ARP: 3).
The results indicate that grape seed meal as byproduct from grape seed oil
extraction could be regarded as potent natural antioxidant additive for food
and as functional dietary food supplement. The radical scavenging properties
of grape seed oil may contribute to its high nutritional value.
50
Factors influencing the bioavailability of flavonoids
Prof. Dr. Siegfried Wolffram, Inst. of Animal Nutrition and Physiology,
Christian-Albrechts-University of Kiel, D-24098 Kiel, Gemany
For a proper understanding of any health-promoting or other biological
activities of dietary flavonoids, a profound knowledge about their
bioavailability is of crucial importance. In a more classical pharmacological
sense, oral bioavailability is principally defined as the amount of a
pharmacologically active substance reaching body tissues after oral
application. This is normally investigated by measuring concentration-time
profiles in blood samples of the mother compound and/or known metabolites
and subsequent mathematical evaluations (pharmacokinetics). This,
however, implies, that the original substance and/or known metabolites are
well characterized with regard to their pharmacological/biological effects. A
more extended view of bioavailability which appears more adequate
regarding nutritional concepts, not only considers tissue concentrations of
the substance under investigation including all active metabolites but also
addresses their biological effects. With respect to the bioavailability of
flavonoids, however, the two major problems are the lack of suitable and
reproducible biomarkers (as indicators for biological effects) as well as
incomplete knowledge about those in vivo metabolites, responsible for the
observed effects.
Flavonoids are subjected to intensive metabolic transformation after oral
intake, including phase I (activation) and phase II (conjugation) reactions.
With only few exceptions, flavonoid aglyca are extensively conjugated with
glucuronic acid and/or sulphate already within the intestinal mucosa and only
these conjugated metabolites are found in the circulation. Whereas our
knowledge regarding the gastrointestinal absorption of flavonoids including
first pass metabolism within the intestinal mucosa and liver has considerably
improved during the past years almost nothing is known about postabsorptive tissue uptake and further conversion to the final effective
metabolites on the cellular level.
After a general description of factors influencing the bioavailability of a
substance, the talk will concentrate on the intestinal absorption of flavonoids
with special emphasis on factors influencing absorption and in turn
bioavailability. As far as available, this will include the chemical forms as
present in natural and processed food, enzymatic transformation within the
GIT by endogenous as well as microbial enzymes, resecretion into the
intestinal lumen by mucosal mechanisms as well as biliary secretion, and the
effects of the food matrix and dietary components, e. g., dietary fat and fibre.
With respect to the flavonoids considered the presentation will mainly deal
with flavanols, flavonols and isoflavones.
19
Prebiotic effects of phyrtochemicals
Ian Rowland, Northern Ireland Centre for Food and Health, University of
Ulster, Coleraine BT52 1SA UK
In the area of functional foods, there is currently considerable interest in
products that may confer health benefits to the consumer by increasing the
number of ‘beneficial’ organisms (usually lactobacilli and bifidobacteria –
lactic acid producing bacteria [LAB]) in the gastrointestinal tract. Food
manufacturers have used two strategies to achieve this. The direct approach
is to supply live preparations of the organisms (‘probiotics’) usually in the
form of fermented dairy products such as yoghurt. One of the perceived
difficulties with probiotics is that after ingestion, a substantial proportion of
them are killed by adverse conditions in the upper gut before they reach the
colon where the main population of bacteria resides. Thus a second, indirect,
approach has been taken in which non-digestible carbohydrate food
supplements are given, which support and stimulate the growth of LAB in the
colonic microflora. These food components have been termed ‘prebiotics’
and include certain polysaccharides such as certain forms of resistant starch
and gum Arabic. The major group of plant derived prebiotics however fall
into the category of low molecular weight carbohydrates called non-digestible
oligosaccharides
(NDO)
e.g.
fructo-oligosaccharides,
galactooligosaccharides and inulin, which are derived from a wide range of plants
including chicory, artichoke, asparagus, onion, leek, soy beans and agave.
Prebiotics have been studied in human trials and in animal models for their
potential beneficial effects on human health. The studies fall into 4 main
areas:
1. Effects on colonic microflora
Prebiotics appear to increase bacterial biomass, which results in a
modest increase in faecal bulk. More importantly, consumption of
prebiotics is often associated with an increase in proportion of LAB in
faeces. The significance of this for human health has not been
established, but in some studies there was a corresponding decrease in
potentially pathogenic genera such as clostridia.
2. Effects on lipid metabolism
Initial studies in rats indicated that inulin feeding was associated with a
hypolipidaemic effect and a lowering of serum triacylglycerols and in
some cases a decrease in total cholesterol. Subsequent studies in
humans however have yielded inconsistent results.
3. Effects on mineral absorption
A number of rat studies have reported that feeding prebiotics improves
the bioavailability of calcium and magnesium. Of the three human
studies conducted, two have provided evidence of an increase in
calcium absorption after feeding NDO.
4. Effects on carcinogenesis
Although there are few human data available, animal studies indicate
that carcinogen-induced tumours and preneoplastic damage to the
colonic mucosa are suppressed by feeding NDO and resistant starch.
Two studies suggest that feeding a combination of probiotic and
prebiotic has a more potent inhibitory effect on preneoplastic lesions
than either product alone.
20
Influence of variety and processing technology on carotenoid
content in pumpkin cultivars
Mirjam Kreck, Research Institute Geisenheim, Petra Kürbel, Michael Ludwig,
Helmut Dietrich; Dep. of Wine Analysis and Beverage Research, Research
Institute Geisenheim; e-mail: [email protected]
Carotenoids are known for a wide range of important and well-documented
biological activities. They act as potent antioxidants, as well as free radical
scavengers, modulate the pathogenesis of cancers as well as coronary heart
disease (1). Various carotenoids, including -carotene, -carotene, and cryptoxanthin, possess provitamin A activity by being transformed into retinal
by mammals. Due to the ability of capturing free oxygen and blue light in the
retina, the xanthophylls lutein and zeaxanthin are also known to provide
protection against macular degeneration connected with aging.
New products are being produced in this line with juice mixtures that provide
increase quality (nutritive value, colour, etc.), such as high carotenoid
content.
Therefore, the carotenoid pigments of different cultivars of Curcubita maxima
(pumpkin) were investigated in pulp, peel and genuine juice by means of
HPLC. In dependence of the variety different concentrations and distributions
of -carotin, -carotin, violaxanthin, neoxanthin, all-trans-lutein, zeaxanthin,
lutheoxanthin and the isomers 9-cis- -carotin and 13-cis- -carotin were
obtained. The total carotenoid content of the pumpkin peel depends on the
variety with 1.2 mg/100g for Butternut up to 175.1 mg/100g for Rouge,
whereas total carotenoid contents in the pulp differed from 1.7 mg/100g for
Baby Bear to 68.3 mg/100g for Rouge. The variety Hokkaido showed high
total carotenoid contents in pulp (21.8 mg/100g) and peel (104.8 mg/100g),
whereas for Butternut minor concentrations in pulp (1.2 mg/100g) and peel
(4.4 mg/100g) were detected.
Additionally, vitamin A values were investigated for pumpkin varieties in
dependence of -carotene (100% activity; 0.6 g -carotene is equivalent to
0.1 RE), -carotene (50% activity) and -cryptoxanthin (50% activity) (3).
Hokkaido appeared to be the best source of provitamin A with average
concentrations of 425 RE/100g in the pulp, 580 RE/100g in the peel and 331
RE/100g relating to the whole vegetable. For the variety Muscat also high RE
values can be calculated with 359 RE/100 g in pulp, 152 RE/100g in peel
and 315 RE/100g calculated for the whole vegetable. The varieties
Bischofsmütze, Butternut and Neon were low in vitamin A value, presenting
averages of 93, 65 and 88 RE per 100 g, respectively.
With regard to pumpkin juice processing, three varieties (Muscat, Rouge,
Neon) were processed in a half technical scale to genuine juices. Additionally
to the carotenoid analysis, inner parameters were investigated in the genuine
juices and pomace extracts.
The red and orange coloured Curcubita maxima varieties are good and
valuable sources of carotenoids among vegetables or vegetable juices.
Kritchevsky, S. B. -carotene, carotenoids and the prevention of coronary heart
disease. J. Nutr. 1999, 129, 5-8.
Murcovic, M.; Mülleder, U.; Neunteufl. Carotenoid Content in Different Varieties of
Pumpkins. J. of Food Comp. And Anal. 2002, 15, 633-638.
Hidaka, T.; Anno T.; Nakatsu, S. The composition and vitamin A value of the
carotenoids of pumpkins different colors. J. Food Biochem. 1987, 11, 59-68.
49
Effect of tomato processing on antioxidant compounds and
antioxidant activity
Do polymorphisms affect bioavailability of plant bioactive
compounds?
Kaufmann K., Ros G., García-Alonso, FJ and Periago MJ
University of Murcia, Dpto. of Food Technology, Food Science and Nutrition;
Murcia, Spain 30071; [email protected]
Paul A Kroon, Institute of Food Research, Colney Lane, Norwich NR4 7UA,
UK. paul.kroon@ bbsrc.ac.uk.
OBJECTIVE
The aim of the present study was to ascertain the effect of thermal
processing on the antioxidant contents (total phenolics, flavonoids,
hydroxycinnamics acids, vitamin C and lycopene) and on the antioxidant
activity of tomato.
METHODOLOGY
Tomato fruits (Lycopersicum esculentum) extra category, size III and bright
red stage, were purchased in the local supermarket. Tomatoes were
homogenised and processing in the Pilot Plant of the Murcia University.
Thermal processing was applied to the raw tomatoes (RT), obtaining two
tomato concentrates: with 10º (TC1) and 15º Brix (TC2). Tomato samples
were placed in air-tight plastic bottles and frozen immediately. Total
phenolics were quantified with Folin-Ciocalteau reagent using gallic acid as
standard, and total flavonoids were measured following the colorimetric
assay described by Zhishen et al. (1999) using quercetin as standard.
Individual flavonoids (rutine, naringenin, quercetin and kaempferol) and
hydroxycinnamic acids (chlorogenic, caffeic and p-coumaric acids) were
determined, after acid hydrolysis, using a HPLC (Crozier et al., 1997;
Martinez-Valverde et al., 2002). Vitamin C content was also detected by
HPLC and lycopene content was determined spectrophotometrically
following the procedure described by Periago et al. (2004). Hydrophilic and
lipophilic antioxidant activity was determined using ABTS method.
RESULTS
The antioxidant content in tomatoes increased significantly after thermal
processing, with the exception of vitamin C. Total phenolics increased from
10.54 to 51 mg/100 g, flavonoids from 0.54 to 4.20 mg/100 and lycopene
from 2.42 to 11.5 mg/100g, for RT and TC2, respectively. These increases
were related to the thermal processing, which reduces the water content and
leads to the rupture of cell structure. Hydroxycinnamic acids also increased,
but no significant effect was observed in flavonoids, with the exception of
rutine content. In addition, the antioxidant activities of both concentrates
were significantly greater than the antioxidant activity of raw samples, and
were correlated with the content of the antioxidant compounds.
Plants contain an array of secondary metabolites that are consumed as part
of human diets. There has been considerable interest in a number of classes
of plant secondary metabolites (e.g. flavonoids, simple phenolics
glucosinolate breakdown products, carotenoids, stilbenes and terpenes)
because they exhibit responses in animal and cellular models that are in
keeping with the protection afforded by diets rich in fruits and vegetables.
The importance of bioavailability with regard to these ‘plant bioactives’ has
become apparent over the last 8-10 years. In particular, it has become clear
that a number of active processes are involved in the absorption and
associated metabolism of some plant bioactives. For example, flavonoids are
present in plants and derived foods and beverages as a mixture of
glycosides, which are hydrolysed during absorption, and appear in plasma
and urine as a number of glucuronide and sulphate conjugates of the parent
aglycone or methylated derivatives. This process requires the involvement of
intestinal enzymes (β-glucosidases, UDP-glucuronosyl-, sulfo- and catecholO-methyl-transferases) and may involve membrane transporters such as the
sodium-dependent glucose-transporter-1 (SGLT-1) and multi-drug resistance
proteins (MRPs). Deglycosylation, transport and conjugation (metabolism)
are also important in the absorption of other plant bioactives including
glucosinolates and other phenolics. A number of these proteins are
polymorphic in humans.
O
OH
O
HO
O
HO
OH
OH
O
OH
Quercetin
OH
O
OH
4'
O
HO 7
Quercetin-3'-sulfate
HO
5
OH
O
O
OH
OH
OH
O
S
O
3
Quercetin-4'-glucoside
OH
O
OH
3'
OH
O
This paper will examine whether polymorphisms affect the bioavailability of
plant bioactives. Two groups of plant bioactives, flavonoids and
glucosinolates, will be used as case studies. Current understanding of the
pathways of absorption and metabolism of these bioactives will be
presented, and the evidence that polymorphisms in metabolic enzymes and
transporters affect bioavailability discussed.
CONCLUSIONS
Thermal processing of tomatoes produces a significant increase of the
antioxidants, with the exception of vitamin C. An improvement of beneficial
effects for human health derived from the tomato consumption can be
achieved during the food processing.
Acknowledgements: “Fundación Séneca/Fondo Social Europeo” of Murcia
Government.
48
21
Apple polyphenols: nutrifunctionnal ingredient and innovative
incorporation possibilities.
Nutrigenomics approach to study the effects of whole grain
products on glucose metabolism and insulin action
Philippe Sanoner, Sabrina Nicolle, Hélène Masson, Tibogo Sanogo, JeanMarie Barré, Val-de-Vire Bioactives, Rte de St Romphaire BP2, 50890
Condé-sur-Vire, Fr. [email protected]
Author: P.Kallio; 1 Food and Health Research Centre, University of Kuopio;
1
2
2
and K.
Co-Authors: M. Kolehmainen , T. Salopuro , L. Pulkkinen
Poutanen1; 1 Food and Health Research Centre, University of Kuopio; 2
Department of Clinical Nutrition, University of Kuopio.
Are governmental programs such as “Eat five a day” or “Programme National
Nutrition Santé” modern equivalents to the old English saying “An apple a
day keeps the doctor away”?
There is abundant evidence, both
1
epidemiological and experimental , that apple polyphenolic antioxidant in the
normal diet can exert protective effects against major metabolic diseases2.
To enrich our diets with apple phytonutriments Val de Vire proposes to
enhance antioxidant capacity of formulations with concentrated apple
polyphenols (AP): Pomactiv®. In the heart of Normandy, the Val de Vire
Company is traditionally brewing cider apples, transforming fruits of a
specific orchard into apple juice and hard cider.
Most of AP are monomeric and polymeric flavan-3-ols, essentially
procyanidins with an epicatechin as constitutive units, followed by
hydroxycinnamic acids (chlorogenic acid mainly), dihydrochalcones
(phloridzin mostly), and flavonols (quercetin glycosides). Nutritional studies
show that AP takes part to the cholesterol lowering capacity of a diet
containing at least 15% apple. AP are able to lower concentrations of
sanguine cholesterol (LDL), decreasing the risk of coronary heart diseases,
as LDL oxidation can lead to arterom plaque formation and vessels
occlusion. This antioxidant effect of AP is also effective as technological
ingredient, for the meat conservation preventing formation of oxidised
cholesterol forms. AP can also be employed for their UV filtering capacities
(UV radiation leading to oxidative damages) for cosmetic applications. In
particular the quercetin, is one major component of the apple UV-screen. AP
exerts also some anti-inflammatory properties, as inflammation involves
oxidative mechanisms. Naturally occurring AP has also physiological effects
on plasma glucose concentrations and circulating gastrointestinal hormone
profiles. The phloridzin, which is currently used in nutritional studies since
1890, contains a glucosyl group in its structure and is a well-known
competitive inhibitor of glucose/Na+ intestinal transporter. Those properties
may be exploited for specifically adapted foods, slowing down glucose
assimilation. AP tanning properties in particular procyanidin conferee to
those compounds large non-specific inhibitory properties, bacteriostatic
activities, which can be helpful for food conservation. For example, apple
procyanidins are able to inhibit the water-insoluble glucans synthesis
responsible for the dental plaque formation and bacterial adherence.
Incorporating AP can bring coloration to the formulated food. But principal
limitation of the integration of polyphenols in formulated foods is the
bitterness brought. Fortunately polyphenols incorporation rates to obtain
nutraceuticals effects are usually low enough to avoid off-taste
developments. Furthermore the AP bitterness can also participate to a new
equilibrated mouthfeel.
Type 2 diabetes (T2DM) and its complications are a significant cause of
morbidity and mortality worldwide. Both genetic and environmental factors
are involved in pathogenesis of T2DM. Whole grain products rich in
polyphenols have been shown to be beneficial on glucose metabolism.
The aim was to study the effects of long-term (12 weeks) use of two wholegrain products (rye bread and oat-wheat bread) as a part of daily diet on
glucose metabolism and gene expression in adipose tissue (AT). Altogether
47 (male/female: 24/23) subjects [age: 55 ± 6y; BMI: 32.1 ± 3.8 kg/m2 ; 2-h pglu: 8.0 ± 2.3 mean ± SD] who had impaired glucose tolerance were
randomized to consume one of the two diets containing either whole grain
rye or whole grain wheat/oat breads (25% of total daily energy) for 12 weeks.
AT biopsies for gene expression analyses were taken before and after the
intervention. Ten subjects from both test groups were chosen for studies
using Affymetrix HG U133 plus 2.0 microarrays, confirmed by Real-Time
PCR.
First phase insulin response improved along the intervention in rye group.
This was shown as an improvement of insulinogenic index (p=0.012) and 30
minutes serum insulin concentration (p=0.036) in oral glucose tolerance test.
According to preliminary data several genes showed changes along the
intervention. Moreover, gene expression was different between the study
groups. Nutrigenomic studies are expected to provide new information of the
mechanisms via which cereals might effect on insulin action in persons with
slightly impaired glucose tolerance and metabolic syndrome.
Williamson G., Manach C. Bioavailability and bioefficacy of polyphenols in humans. II.
Review of 93 intervention studies. Am. J. Clinical Nutrition. 2005, 81, 1, 243S255S.
Boyer J. and Liu RH. Apple phytochemicals and their health benefits.Nutrition Journal
2004, 3:5
22
47
Studies on anticomplementary activity of ethanol extracts from
pumpkin
How do we market Phytochemically rich products- public health
versus consumer needs
He Hongju1, Che Ruixiang2, Chen Guilin2, Li Haizhen1; 1National Engineering
2
Research Center for Vegetable, Beijing 100089, China; Horticulture College,
Hebei Agriculture University, Baoding 071001, China; e-mail:
[email protected]
Professor Aedin Cassidy School of Medicine, Health Policy & Practice
University of East Anglia, Norwich NR4 7TJ, UK
The complement system is known to play a major role in inflammation
activation. Complement is a series of proteins found in blood serum that
attack bacterial cells or other foreign cells, causing lysis of cellular
constituents as a result of damage to the cell membrane (Kuby 1997). These
proteins play a critical role in aiding phagocytosis of immune complexes and
are critical in the regulation of cellular immune responsiveness (Kimball
1990). Thus, the human complement system plays an important role in the
host defense system, and activation of the system contributes to pathologic
reactions in a variety of inflammatory and degenerative diseases (Walpott
1996).
Cucurbita spp. is an edible annual plant and divided into 3 groups: Cucurbita
moschata Duch, Cucurbita maximum Duch, and Cucurbita pepo L. Among
them, Cucurbita moschata Duch (Pumpkin) is dominant in China. Pumpkin
has plenty of nutritional ingredients and physiologically active substances
and is widely used as a traditional medicine against various diseases, as well
as a popular food source. Although it has pharmacological effects as well as
a nutritional value, little knowledge is available on the active compounds
having medicinal effect. Therefore, to elucidate the pharmacological effects
of Cucurbita moschata Duch, anticomplementary activities were examined.
The pumpkin fruit of twenty cultivars was studied for anticomplementary
activity of ethanol extracts and total soluble sugar assessed by the CH50
method. The results showed that different pumpkin cultivars have different
anticomplementary activities of ethanol extracts ranged from 67.9% (cv.
Balenggua) to 80.6% (cv. Jingyinli). The anticomplementary activity of
ethanol extracts for the same pumpkin cultivar was higher than the
anticomplementary activity of total soluble sugar. But there is no correction
between the activity and sugar content.
Although there is convincing evidence that plant based foods prevent the risk
of chronic disease, teasing out the relative contribution that specific
phytochemicals make to this reduction in disease risk is currently unclear.
For some Phytochemicals there is significant evidence that they are
biologically active in vitro, but for human health it is important to define the
bioavailability of these compounds, determine levels attained in target
tissues following consumption and elucidate biological effects in humans.
Lessons learned from antioxidant vitamin research suggest we should be
cautious about removing the perceived ‘active’ component from plant based
foods and promoting consumption as isolated supplements without clear
scientific evidence from human studies that this preparation is quality
assured and proven to be as effect as a food based diet.
Kuby J. 1997. Immunology. 3rd ed. New York: Freeman. P 371-393
Kimball JW. 1990. Introduction to immunology. New York: McMillan. P 331-350
Walpott M. 1996. Complement. In: Roitt I., Broscoff J, Male, D, editors. Imunology. 4th
ed. London: Mosby. p 13.1-13.12.
46
23
Glucosinolates composition and variation in Brassica rapa
Hongju He1,2,
Lou Ping2, Ruud Verkerk2, Matthijs Dekker2, Guusje
2 1
Bonnema ; National Engineering Research Center for Vegetables, Beijing
2
Wageningen University, 6700 AJ Wageningen, The
100089, China;
Netherlands; e-mail: [email protected]
Abstracts
Short Communications
In chronological order
Glucosinolates (GSs) are a diverse class of S- and N-containing secondary
metabolites that are mainly found in members of the Brassicaceae.
Glucosinolate breakdown products are proposed to act as allelochemicals
and to play a role in plant defenses and human health. (Verkerk and Dekker,
2004; Dekker et al., 2000). Glucosinolate distribution and content of most
widely consumed Brassica vegetables such as cabbage, cauliflower, brussel
sprouts and broccoli were reported (Mithen et al., 2000). While many reports
exist on the occurrence of glucosinolates in Brassica oleracea, less is known
on their occurrence in Brassica rapa.
Brassica rapa (2n=20) contains various cultivars specialized for oil-seed
production. In addition, it is also used as a vegetable, for either leaves or
stems are harvested. The main focus of the KNAW-MOST Brassica
nutrigenomics program”Genomics of micro-nutrition traits in Brassica rapa for
improved human health” is the exploitation of existing variation in B. rapa
germplasm using molecular genetic tools. The primary focus will be
genomics of glucosinolates in Brassica rapa for improved human health.
Eight different kinds of glucosinolates were identified in the Brassica rapa
group (progoitrin, gluconapin, glucobrassicanapin, gluconasturtiin, 4OH
glucobrassicin,
Glucobrassicin,
4ME
glucobrassicin
and
NEO
glucobrassicin). Gluconapin, gluobrassicanapin and gluconasturtiin are the
most dominant GSs in Brassica rapa group, but there are much more
variations among species and cultivars.
Total glucosinolate contents in Brassica rapa group varied substantially
between the different species. The highest amount of GSs (1521 µmol/ 100g
FW) was observed in vegetable turnip leaves, followed by yellow sarson and
rapid cycling, respectively 379 and 253 µmol/ 100g FW. Whereas the lowest
GSs content was found in pak choi (72 µmol/ 100g FW). Relatively high
amounts of gluconapin (840 µmol/ 100g FW) and glucobrassicanapin (634
µmol/ 100g FW) were observed in the leaves of vegetable turnip.
Verkerk R. and M. Dekker (2004) Glucosinolates and myrosinase activity in red
cabbage (Brassica oleracea L. var. Capitata f. rubra DC.) after various microwave
treatments. Journal of Agricultural and Food Chemistry, 52, 7318-7323
Dekker M., R. Verkerk and W.M.F. Jongen (2000) Predictive modelling of health
Trends in Food Science, 11(4/5), 174-181
Mithen R.F.; Dekker M.; Verkerk R.; Rabot S.; Johnson I.T.; (2000). Review: The
nutritional significance, biosynthesis and bioavaiability of glucosinolates in human
foods. Journal of the Science of Food and Agriculture, 80, 967-984
24
45
Induction of flavonoid biosynthesis in the model plant
Arabidopsis thaliana by various pre-harvest conditions
Karena Hoffmann-Wülfing, Wolfgang Bilger; Plant Ecophysiology, Botanical
Institute, University of Kiel, Am Botanischen Garten 1-9, 24118 Kiel,
Germany
Flavonoids are a group of phenolic compounds with potential beneficial
impact on human health due to their antioxidative, anticancerogenic and
antimicrobial properties in vitro. Among this diverse group quercetin is the
most promising phenylpropanoid as it showed the highest antioxidative
capacity. Hence, it would be desirable to produce plant food with an
increased quercetin content.
UV-B has been reported to induce mainly quercetin biosynthesis. It has also
been shown that low temperature induces flavonoid biosynthesis in general.
Therefore, we asked which conditions optimise quercetin formation.
Arabidopsis thaliana is a model plant for species of the mustard family which
comprises food plants like cabbage (Brassica sp.), mustard (Sinapis sp.),
rocket (Eruca sp.) and others. Since the Arabidopsis genome is fully
sequenced and a large assortment of mutants and transgenic plants
available, this species is ideal to investigate the regulation of flavonoid
biosynthesis by growth conditions.
Flavonoids accumulate predominantly in epidermal vacuoles where they
screen the mesopyll of leaves against harmful UV-B and UV-A radiation.
These characteristics allow us to estimate flavonoid content by measuring
epidermal UV-A transmission in green leaves using a newly developed
technique based on chlorophyll fluorescence (Bilger, Johnsen & Schreiber,
2001). This method has the advantage of being rapid and non-invasive. We
also applied HPLC analysis in order to investigate the flavonoid composition
under different pre-harvest conditions.
Preliminary results indicate that under moderate UV-B, high light or low
temperature not only quercetin derivatives are accumulated but also
kaempferol derivatives. However, experiments with different light qualities
strongly suggest quercetin formation being exclusively under blue or UV light
control.
Accordingly, growth at moderately low temperature and moderate blue light
intensities for several days can enhance quercetin content in leaf tissue as
good as UV-B irradiation.
Production of health related flavonoids in tomato fruit
Elio Schijlen, Ric de Vos, Harry Jonker, Hetty van den Broeck, Jos Molthoff,
Arjen van Tunen and Arnaud Bovy, Plant Research International,
Wageningen, The Netherlands. [email protected]
Flavonoids comprise a large and diverse group of polyphenolic plant
secondary metabolites. In plants, flavonoids play important roles in many
biological processes such as pigmentation of flowers, fruits and vegetables,
plant-pathogen interactions, fertility and protection against UV light.
Being natural plant compounds, flavonoids are an integral part of the human
diet. Nowadays, there is increasing evidence that dietary polyphenols are
likely candidates for the observed beneficial effects of a diet rich in fruits and
vegetables on the prevention of chronic diseases.
To date, many structural genes as well as transcription factor genes involved
in the flavonoid biosynthetic pathway of different plant species have been
isolated and characterized. We have used some of these genes to
genetically engineer the flavonoid pathway of tomato in order to explore the
possibilities for developing food products with increased nutritional value.
We used three different approaches to engineer the flavonoid pathway in
tomato (Figure 1). I) Increasing endogenous tomato flavonoids. Highly
increased levels of peel flavonols were obtained by overexpressing the gene
encoding CHI, the rate-limiting enzyme of the flavonoid biosynthesis pathway
in tomato. Furthermore, flavonoid accumulation in the fruit flesh resulted from
the overexpression of the transciption factor genes Lc and C1. II) Blocking
specific steps in the flavonoid biosynthesis by RNA interference. III)
Production of novel tomato flavonoids by engineering the biosynthetic
pathway at three different steps: 1) Introduction of a stilbene synthase
encoding gene resulted in the accumulation of stilbenes. 2) Deoxychalcones,
normally restricted to legume crops, were produced by the combined
overexpression of chalcone synthase and chalcone reductase. 3) Finally,
tomatoes with high levels of flavonols and flavones were obtained after
enhancing the endogenous pathway combined with the introduction of a new
branch leading to flavones.
Using an on-line HPLC antioxidant detection system, we demonstrated that
the novel flavonoids present in the transgenic tomato fruits displayed good
antioxidant activities. The results of our genetic engineering work
demonstrate the possibilities to change the levels and composition of health
related flavonoids in tomato fruits and provide more insight in the genetic and
biochemical regulation of the flavonoid pathway. In addition, the engineered
tomatoes are excellent material to study the potential health effects of dietary
flavonoids present in a common food matrix.
Flavonols
Transcription factors Lc C1
FLS
CHS
Malonyl-CoA
F3H
CHI
Chalcones
Flavanones
Dihydroflavonols
Phenylpropanoids
CHS+CHR
FNS
DFR
ANS
STS
Stilbenes
Deoxychalcones
Flavones
Anthocyanins
44
Figure 1. Schematic of flavonoid pathway engineering in tomato
Solid black arrows represent the active pathway in tomato fruit peel. Anthocyanin production is occasionally found in
vegetative tissues (dotted arrow). Increased flavonol synthesis was obtained by enhancing the pathway by overexpressing Chi
or by introducing the transcription factors Lc and C1 (blue arrows). RNAi approach was used to obtain fruits with (partially)
blocked flavonoid synthesis (red crosses). New introduced flavonoid branches are represented by green arrows
25
Postharvest radiation treatment to increase health-related
quality of fruit and vegetables
a
Gunnar Bengtsson
, Sidsel Fiskaa
Hagenab, Knut Asbjørn
Solhaugb, Roman
a
a
a
Schöner , Emanuele Lombardo , Jennifer Schöner
, Grethe Iren Borgea,
c a
Arvid Bergeb, Karin Haffnerb & Wolfgang Bilger
;
MATFORSK
– Norwegian
Food Research Institute, Aas, Norway, b Norwegian University of Life
Sciences, Department of Ecology and Natural Resource Management, and
Department of Plant and Environmental Sciences, Aas, Norway; ,c University
of Kiel, Kiel, Germany
Visible light is necessary for chlorophyll production, photosynthesis and a
normal development of green plants. Ultraviolet (UV) radiation from the sun
can damage living cells, and therefore, there is a defence mechanism in
plants where UV radiation induces the formation of UV absorbing flavonoids
in the epidermis. In addition to colourless flavonoids such as flavones and
flavonols, anthocyanins with a red colour are also synthesised in response to
the sun. It has been earlier shown that flavonoids and chlorogenic acid can
be induced postharvest in apple skin. However, the effect of postharvest
radiation on product quality in general has been investigated in fresh fruit and
vegetables to a very limited extent. As examples we have studied the effects
of post-harvest radiation treatments on chemical parameters of apples and
broccoli related to human health.
Norwegian grown broccoli (cv. Marathon) or apples (cv. Aroma) were treated
part of the day for up to 12 days with various combinations of visible light and
UV-A and UV-B radiation in controlled climate rooms. Ascorbic acid,
chlorogenic acid and flavonoids were quantitated by HPLC, and total
phenols, soluble solids and titratable acidity by other methods. Total
antioxidant capacity was measured in methanol extracts by the ORAC
(Oxygen Radical Absorbance Capacity) method.
The results show that post-harvest treatment with UV-B and visual light
increased the content of UV absorbing flavonoids, anthocyanins, chlorogenic
acid and ascorbic acid in apples. The effect was strongest in suboptimal
(green) apples harvested from the inner tree canopy. The light treatments
had no influence on the content of soluble solids or titratable acidity in any of
the apples.
In broccoli, the levels of flavonoids and total antioxidant capacity were much
higher in flower buds than in stalks of the inflorescence or in the stem.
Flavonoid levels in flower buds tended to increase after treatment with visual
light + UV-A + UV-B, but the increase was not statistically significant.
Radiation treatment did not change the total antioxidant capacity. The natural
variation between broccoli plants was very large: for flavonoid content tenfold
and for the ORAC value more than twofold in dark green flower buds. This
variation was not reduced by the post-harvest radiation treatment.
The present study shows that it is possible to increase not only flavonoids
but also other health-related constituents in fruit and vegetables by postharvest radiation treatment. This was demonstrated for apples. The broccoli
experiment was probably carried out at sub-optimal conditions and the
number of constituents analysed was too small for us to draw any final
conclusion. The fact that controlled radiation treatment post-harvest can
change the health-related quality is an indication that the incident light during
distribution of fruit and vegetables also could have an effect, which might be
positive or negative depending upon the conditions.
26
Identification of Phenolic Compounds in Leaves of Chinese
Brassica Vegetables (Brassica campestris L. ssp. chinensis var.
communis) by HPLC-ESI/MS
B. Harbaum, E. Hubbermann, H. Stöckmann, K. Schwarz; Institute of Human
Nutrition and Food Science, University of Kiel, Germany
Brassica vegetables are known to play an important role in human nutrition
due to their content of phytochemicals such as vitamins, minerals and
phenolic compounds. Several studies have investigated the phenolic
composition of members of the Brassica family like cabbage. However,
information about the phenolic composition of Chinese Brassica vegetables
is scarce. Shakakibara et al. (2003) found that kaempferol is the main
flavonoid in Chinese Brassica vegetables like Pak-Choi. However, cabbages
and broccoli also contain luteolin, quercetin and apigenin derivatives
(Bahorun et al., 2004).
Pak Choi (Brassica campestris L. ssp. chinensis var. communis) was
cultivated in a green house in summer 2005. Eight week old plants were
harvested and the leaves were freeze dried. Extraction was facilitated by
ultrasonic treatment using 50 % methanolic solution. The extract was diluted
with water and analysed by reversed phase HPLC (C18 column;
water/acetonitrile/formic acid, gradient elution). The detection and
identification was carried out by UV (330nm) and mass spectrometry
including MSn fragmentation.
The main flavonoids identified in Pak Choi were kaempferol derivatives.
Kaempferol was found to be esterified with two, three or four hexoses and
acylated with different hydroxycinnamic acids. Furthermore, different
hydroxycinnamic acid derivatives were detected. In addition to the identified
quinic acid derivatives such as monocaffeoyl- and coumarylquinic acids other
hydroxycinnamic esters were characterized which are presumed to be esters
of malic acid.
Bahorun, T., Luximon-Ramma, A., Crozier, A., Aruoma, O. (2004) Total phenol,
flavonoid, proanthocyanidin and vitamin C levels and antioxidant activities of
Mauritian vegetables, Journal of the Science of Food and Agriculture 84: 15531561
Sakakibara, H., Honda, Y., Nakagawa, S., Ashida, H., Kanazawa, K. (2003)
Simultaneous Determination of all Polyphenols in Vegetables, Fruits, and Teas,
Journal of Agriculture and Food Chemistry 51: 571-581
43
Content and antioxidative capacity of carotenoids, -tocopherol
and polyphenolics in compartiments from tomatoes grown
under saline conditions
Graßmann, J., Wilfried H. Schnitzler, Markus Woitke, Sandra Krauß; Institute
of Vegetable Science, Life Science Center Weihenstephan, TUM, Dürnast 2,
85350 Freising
Saline growing conditions influence the inner quality of tomato fruit. As part
of it the content of carotenoids and their antioxidative capacity is significantly
enhanced by rising salinity levels. Our aim was to investigate to what extent
this increase can be attributed to different compartments of the fruit
quantitatively, i.e. to the peel (epidermal structure / exocarp) and the pulpae
(endo – and mesocarp).
Tomatoes of the cultivar ‘Durinta’ were grown in a recirculating hydroponic
system under three salinity levels (EC 3, 6.5 and 10), whereas the basic
nutrient solution comprised EC 3 (control) with addition of NaCl to reach EC
6.5 and 10. Tomatoes were harvested in fully ripe stage, peel and pulpae
were separated immediately after harvest, frozen, lyophilized and ground.
Content of carotenoids,
-tocopherol and polyphenolics and resulting
antioxidative capacity was analyzed by HPLC, Folin-test and the ABTSsystem, respectively.
We could show that the antioxidative capacity is significantly higher in the
peel compared to the flesh and that rising salinity lead to increasing AC in
the peel both for carotenoids and polyphenolics. Regarding the pulpae, only
the highest salinity level improved AC. Likewise, analysis of the most
important lipid-soluble antioxidants, i.e.
-carotene, lycopene and
tocopherol revealed increasing concentrations. This increase however was
much more pronounced in the peel compared to the fruit. Examination of the
polphenolic content by means of Folin-Ciocalteu revealed only slight
increases in the flesh/pulp with rising salinity, in the peel the polyphenolic
content increased stronger.
The found higher AC on a fresh weight basis can be attributed to a
concentration effect in terms of higher dry matter at higher salinity levels. The
higher content of antioxidants in the epidermal structure in comparison to the
endo- and mesocarp marks their importance as a protective boundary layer
towards environmental impacts. An increased synthesis of carotenoids and
other protective compounds may account for a higher sensitivity of the plants
to salinity stress.
Effect of enzymatic processing on anthocyanins in bilberry and
black currant juice
Jani Koponen1, Riitta Törrönen1, Annikka Mustranta2, Kaisa Poutanen1,2 and
2 1
Johanna Buchert , Food and Health Research Centre, University of Kuopio,
Kuopio, 2VTT Biotechnology, VTT, Finland
Black currants (Ribes nigrum) and wild bilberries (Vaccinium myrtillus) are
commonly used in juice manufacture in Scandinavian countries. Traditional
processing methods may result in low juice yield and incomplete extraction of
potential bioactive components such as anthocyanins. Therefore, enzymes
are currently used to facilitate the processability of berries.
Anthocyanins are the major and important group of phenolic compounds
present in black currants and bilberries, and are mostly responsible for the
red and blue colours of these berries. Moreover anthocyanins are expected
to have health promoting characteristics, because of their high antioxidant
activity. However, the effects of enzymatic processing on these bioactive
compounds are poorly known.
In this study, the effect of enzyme-aided processing on anthocyanins in
bilberry and black currant juices were studied using three different doses of
commercial cell wall degrading enzyme preparations. One cellulase
(Econase CE) and three pectinase (Biopectinase CCM, Pectinex Smash XXL
and Pectinex BE 3-L) preparations were dosed based on their
polygalacturonase activity. The juice yields were measured by weighing, and
contents of total anthocyanidins, including aglycons and anthocyanins, were
determined by high performance liquid chromatography (HPLC-DAD). The
yield of total anthocyanidins from bilberries and black currants was
calculated using the measured values.
Enzyme-aided processing increased the juice yield by 13 – 25 % and 8 – 36
% in bilberries and black currants, respectively. The content of total
anthocyanidins in enzyme-treated bilberry and black currant juices were
2970 – 5140 mg/kg and 2870 – 3330 mg/kg, respectively. The highest
concentration of total anthocyanidins was achieved with Pectinex Smash
XXL, which increased the content by 46 % and 19 % in bilberry and black
currant juices, respectively. In addition the yield of total anthocyanidins was
increased by up to 83 % and 58 % in enzyme-treated bilberry and black
currant juices, respectively, as compared to non-enzymatically processed
juices. This increase was clearly dependent on enzyme dose.
These results show that enzyme preparations used in the production of berry
juices improve the juice yield and the extraction of anthocyanins into the
juice. Thus, enzyme-aided processing offers a powerful tool for improving the
health value of berry juices.
This study has been carried out with the financial support from the Commission of the
European Communities, specific RTD programme “Quality of Life and Management of
Living Resources”, proposal number QLK1-CT-2002-02364 “Novel enzyme-aided
extraction technologies for maximized yield and functionality of bioactive components
in consumer products and ingredients from by-products”, acronym Maxfun. It does not
reflect its views and in no way anticipates the Commission’s future policy in this area.
42
27
Modelling the consequences of variability in food production
chains on human health
Fruit quality of tomatoes grown in a simplified recirculating
soilless system under saline conditions
Matthijs Dekker and Ruud Verkerk, Wageningen University, Product Design
and
Quality
Management
Group,
The
Netherlands.
e-mail:
[email protected]
Graßmann, J., Wilfried H. Schnitzler, Markus Woitke, Sandra Krauß; Institute
of Vegetable Science, Life Science Center Weihenstephan, TUM, Dürnast 2,
85350 Freising
Many steps in the food production chain of fruit and vegetable products can
have a large impact on the final human intake of health protective
components. The concept of predictive modelling of health aspects in the
production chain of fruit and vegetable products has been proposed to deal
with this variability (Dekker et al., 2000). In epidemiological studies this
variation in the content of phytochemicals in similar products is usually not
taken into account. In this presentation the effects of various steps in the
production chain on the level of glucosinolates will be presented. This
information is used in a case study to determine the consequences of this
variation on the predictive power of epidemiological studies. Monte Carlo
simulations are used as a tool to predict the effects of the variability in food
production chains on the outcome of epidemiological studies (Dekker and
Verkerk, 2003).
Irrigation with saline water affects tomato fruit quality. While total fruit yield
decreases with salinity, inner quality characterized by taste and health
promoting compounds can be improved.
The variation of glucosinolate levels throughout the chain has been
determined in a quantitative way. Effects of cultivar, storage, industrial
(thermal) processing and domestic cooking have been studied. With this
information it was predicted that large health protecting effects of the
phytochemicals would not be reproducibly picked up by epidemiological
studies as they are currently designed (Dekker and Verkerk, 2005). The
effects of other study designs can easily be simulated by the approach
presented here. The development of predictive models to predict the effect of
the conditions during a thermal treatment (blanching, canning or cooking) of
the product on the level of phytochemicals will be presented. Using product
and consumer information in these predictive models to calculate the effect
on the level of the phytochemicals can lead to a more efficient design of
epidemiological studies, given the practical and financial limitations that will
always be present.
For a detailed description of this relationship the influence of three different
salt levels (EC 3, 6.5 and 10) in hydroponically grown tomatoes was
investigated. Rising salinity levels in the nutrient solution significantly
increased vitamin C, lycopene, β-carotene and flavonoids in fresh fruits up to
35%. The phenol concentration was tendentiously enhanced and the
antioxidative capacity of phenols, flavonoids and carotenoids increased on
fresh weight basis. Additionally, the higher EC values caused an increase of
total soluble solids and organic acids, parameters determining the taste of
tomatoes. Total fruit yield, single fruit weight and firmness significantly
decreased with rising EC levels.
As all desirable characteristics in the fresh produced tomato increased when
exposed to salinity, salinity itself constitutes an alternative method of quality
improvement. Moreover it can compensate for the loss of yield by the higher
inner quality due to changing demands by the market and the consumer.
Other important areas that can benefit of this approach are product and
process design and consumer advice with regard to the preparation of foods
containing certain health-promoting phytochemicals.
M. Dekker, R. Verkerk and W.M.F. Jongen (2000) Predictive modelling of health
Trends in Food Science, 11(4/5), 174-181.
M. Dekker and R. Verkerk (2003) Dealing with Variability in Food Production Chains: A
Tool to Enhance the Sensitivity of Epidemiological Studies on Phytochemicals,
Eur. J. Nutri.,42, 67-72.
M. Dekker and R. Verkerk (2005) Re: Fruit and Vegetable Intake and Risk of Major
Chronic Disease, Journal of the National Cancer Institute, 97(8), 607-608.
28
41
Cytoprotective activity of a phenolic-rich juice in Hepg2 cells
1
1
2
1 1
García-Alonso FJ , Periago MJ , Vidal-Guevara ML , G Ros ; University of
Murcia. Dpt. of Food Technology, Food Science and Nutrition, Spain; 2Hero
España, S.A., Department of Research and Development, Spain.
[email protected]
BACKGROUND AND AIMS
In addition to vitamins and provitamins, phenolics are suggested to be the
major bioactive compounds for the health benefits provided by fruits and
vegetables. A variety of fruit and vegetable extracts have been found to
protect against oxidant-induced damage, in cell-culture models. The aim of
the present study was to evaluate the ability of a functional juice rich in
polyphenols (3 g/L) to protect HepG2 cells (human hepatocyte carcinoma)
from tert-butylhydroperoxide (tB-OOH) and hydrogen peroxide (H2O2)induced oxidative stress.
METHODOLOGY
For the in vitro cell-line experiments, cells were preincubated with juice
extracts for 3 hours and later exposed to tB-OOH or H2O2. After the
treatments, cells were analyzed for cytotoxicity and lipid peroxidation. Cells
exposed to tB-OOH were also analyzed for glutathione peroxidase (GPx)
and glutathione-s-transferase (GST) activities. The radical scavenger N,N’diphenyl-1,4-phenylene-diamine (DPPD) and the intracellular iron chelator ophenanthroline (o-phe), were also assayed in order to elucidate the
mechanisms of action of juice phenolics.
RESULTS
Cells preincubated with juice extracts showed increased resistance to
oxidative challenge, as revealed by an increase in cell survival and a
decrease in lipid peroxidation after exposure to both H2O2 and tB-OOH. Both
DPPD and o-phe protected from lipid peroxidation caused by either H2O2 or
tB-OOH. Detoxification of tB-OOH by cells caused an increase in both GPx
and GST activity. Both juice and o-phe decreased GPx activity, but not that
of GST. The tB-OOH-induced GST activation was only suppressed by
DPPD.
Red wine alcohol promotes quercetin absorption and directs its
metabolism towards isorhamnetin and tamarixetin in rat
intestine in vitro
a
Stefania
a
Dragoni, bJennifer
Gee, bRichard Bennett, aMassimo Valoti and
a
Giampietro Sgaragli; Dipartimento di Scienze Biomediche, Università di
Siena, Via Aldo Moro 2, 53100 Siena, Italy. bInstitute of Food Research,
Norwich Research Park, Colney, Norwich, NR4 7UA, UK
The aim of this study was to investigate the effects of alcohol on the
intestinal absorption of quercetin and its 3-O-glucoside present in red wine
using the rat cannulated everted jejunal sac technique. Tissue preparations
were incubated in whole or dealcoholised red wine, diluted 1:1 with Krebs
buffer for twenty minutes at 37°C, after which the mucosa was removed and
processed for HPLC analysis. Tissues exposed to whole red wine solution
showed significantly higher amounts of both quercetin (x3) and quercetin-3O-glucoside (x1.5), as compared to tissues incubated with dealcoholised red
wine solution. In addition, the tamarixetin and isorhamnetin content of
mucosal tissues incubated with whole red wine solution were approximately
220% and 170% greater than those found after incubation with the
dealcoholised wine solution. When everted rat jejunal sacs were incubated
with Krebs buffer containing a mixture of pure quercetin andquercetin-3-Oglucoside with or without alcohol, tissues incubated in the presence of
alcohol had a significantly higher quercetin content and a trend towards
higher levels of quercetin-3-O-glucuronide and isorhamnetin. In conclusion,
the present findings demonstrate for the first time the facilitating action of
ethyl alcohol in the gut absorption of quercetin. It would appear that the
moderate alcohol content in red wine contributes to its beneficial health
effects in humans by both increasing the absorption of quercetin and
quercetin-3-O-glucoside and by stimulating the formation of O-methylated
metabolites of quercetin, isorhamnetin and tamarixetin, , which are believed
to play a role in protection against cancer and cardiovascular diseases.
CONCLUSIONS
After a short-term preincubation period, phenolics in the functional juice
protected cells from oxidant-induced oxidative stress. This therefore
indicates that phenolics were rapidly absorbed by cells and contributed to the
cellular antioxidant defences. Data also suggest that the protective activity of
the phenolics in the functional juice may partly reside in its iron-chelating
ability.
Acknowledgements:
Government of Murcia.
40
Fundación
Séneca/Fondo
Social
Europeo.
29
Bioavailability of the flavonol quercetin is influenced by the
dietary fatty acid pattern
Iron, zinc and calcium availability from infant cereals with or
without dephytinization.
Stephanie Lesser, Rainer Cermak, Siegfried Wolffram; Institute of Animal
Nutrition and Physiology, Christian-Albrechts-University Kiel, D-24098 Kiel,
Germany. [email protected]
Frontela, C.; Haro, JF.; Romero, F; Ros, G.; Martínez, C.
Bioavailability of flavonols was found to be influenced by the composition of
the diet.
In a previous study, we investigated the influence of dietary fat (lard) on the
oral bioavailability of quercetin in the pig. We observed that an increase in
dietary fat content from 3 to 17% (wt/wt) significantly enhanced quercetin
bioavailability with no further effect on flavonol bioavailability when the fat
content was increased to 32% (Lesser et al., 2004, J Nutr 134). Transferred
to human nutrition, 17% crude fat content (i.e. 34 energy%) is in the range
recommended by the German Nutrition Society for dietary fat intake. We
speculated whether dietary medium-chain triglycerides (MCT), that do not
require chylomicron formation during intestinal absorption, would exert the
same enhancing effect on flavonol bioavailability as long-chain triglycerides
(LCT) used in the above mentioned study.
Six growing male castrated pigs (~35 kg body weight) were surgically
equipped with permanent catheters in the jugular vein. They were fed a
commercial pig diet (designated as standard diet) based on wheat, defatted
soy bean meal and barley with a crude fat content of 2%. Quercetin (30
µmol/kg body weight) was administered in test meals either consisting of 200
g of the standard diet or of isoenergetic portions of standard diet enriched
with 15 g fat/100 g diet (either MCT oil or lard as a source for LCT). After
ingestion of the test meals, blood samples were repeatedly drawn over a
period of 24 hours. Plasma flavonol content was analysed by HPLC after
treatment of samples with β-glucuronidase/sulfatase. In addition, the
influence of the different diets on gastric emptying was investigated in rats
with a body weight of ~250 g. Rats (n=6 per diet group) were trained to
consume 5 g of the standard, LCT or MCT diet within 15 min. After 60 min,
the animals were killed and the dry matter content of the stomach was
determined.
Quercetin bioavailability was enhanced by 38 (P <0.05) and 12 % with the
MCT and LCT diets, respectively, compared to the standard diet. Maximal
plasma levels of quercetin were reached significantly later with the MCT diet
(tmax = ~170 min) than after intake with LCT or standard diet (tmax = ~65 and
95 min, respectively). There was no difference in gastric dry matter content
between the three diets in rats.
In summary, administration of the flavonol quercetin together with a diet
containing MCT fat enhances the bioavailability of the flavonol. Interestingly,
absorption of quercetin was significantly delayed as indicated by the late
maximal plasma levels. However, this was not due to a slower gastric
emptying of the MCT diet. The reason for this finding has to be clarified in
future studies.
This study was financially supported by the German Research Foundation
(DFG). SL is a scholar of the Research Training Group 820 of the DFG.
30
Cereals, incorporated as flours, are usually the first solid foods given to infant
to supplement breast milk and infant formula, coinciding with a period of
rapid growth and development when an adequate diet is crucial to meet
nutrients needs (COMA, 1995). The main minerals supplied by cereals are
calcium, iron and zinc; however, cereals are also a source of antinutrients
like phytic acid, which can negatively affect mineral bioavailability due to its
strong ability to chelate multivalent metal ions (Davidsson et al, 1997). The
influence of phytic acid on iron and zinc absorption is of great public health
importance (Hurrell, 2004).
Our aim was to study the effect of a microbial phytase on mineral
bioavailability of different infant cereals (8 cereals-honey, multicereals, gluten
free, rice cream and wheat) by in vitro methods based on simulation of
gastrointestinal digestion and estimation of the proportion of the ingested
mineral convertible to an absorbable form by measuring the fraction that
dialyses through a membrane. The dialysability of a mineral gives an
estimation of its availability for absorption (Sebastiá et al, 2001). We have
also estimated the influence of water or infant formula addition to infant
cereals with or without dephytinization, on mineral availability. Mineral
dialisability was determined according to Miller at al, (1981).
For that, infant cereals were homogenized in water or infant formula following
manufacturers instructions, and later, treated or not with phytase (10 mg/100
g of homogenate) in order to remove completely the phytic acid content. The
homogenates were exposed to pepsin at pH 2. NaHCO3 was used to adjust
the pH to intestinal levels and digestion was continued after the addition of
pancreatin and bile salts. Minerals from the digestion mixture which diffused
across a semipermeable membrane inside a dialysis bag were used as an
indicator of available mineral. The dialysis bag content was removed and its
mineral
concentration
was
measured
by
atomic
absorption
spectrophotometry.
Mineral dialysability was calculated from the amount of each dialysed
mineral, expressed as a percentage of the total amount present (Miller,
1981).in the homogenates (infant cereal + infant formula or water).
Iron and zinc availability were statistically higher (p<0,05), in infant cereals
treated with phytase while calcium dialysability showed no changes due to
the enzymatic treatment. Complete degradation of phytate increased iron
availability in a range of 0,25% (multicereals) to 2,7% (8 cereals honey), and
from 0,39% (rice cream) to 4,52% (multicereals) in zinc availability.
Although infant formula added to infant cereals increased mineral
concentration of homogenates, the percentages of dialisated iron, zinc and
calcium were similar (p<0,05) to infant cereals mixed with water.
39
Flora: Flavonoids and related phenolics for healthy living using
orally recommended antioxidant
E. Butelli and C. Martin
There is growing evidence that bioactives in the diet play an important role in
promoting health. Flavonoids and related phenolics are examples of
bioactives from plants that have beneficial influences on a number of
important risk factors associated with cardiovascular disease (the leading
cause of death in Europe), cancer and age-related degenerative diseases.
The aim of the FLORA project is to establish Europe at the forefront in
understanding the link between diet and health, particularly the beneficial
effects of flavonoids and related phenolics on cardiovascular disease, stroke
and cancer.
This will be done by fostering a unique interdisciplinary approach between
plant geneticists, plant biochemists, chemists, food technologists, medical
pharmacologists and clinical epidemiologists in this project.
By providing clear recommendations of the efficacy of particular flavonoids
and related phenolics to confer protection against target diseases and
reliable estimates of active phenolic levels in a directory of foods, this
programme will result in a recipe for promoting health through diet that will
appeal to the diversity of tastes of European consumers.
38
Urinary excretion of antioxidants and phenolics following fruit
juice intake - influence of storing.
1
1
1
2
Roland Bitsch , Michael Netzel , Gabriele Netzel ,Christine Thielen ,Helmut
2
3
3
Dietmar-Rolf
Kammerer ,
Andreas
Schieber ,Reinhold
Dietrich ,
Carle3,Thomas Frank4,Undine Ott5,Irmgard Bitsch6; 1Institute of Nutritional
2
Sciences, Jena University,D-07743Jena, Institute of Enology and Beverage
Research, Research Institute Geisenheim, D-65366Geisenheim, 3Institue of
Food Technology Section Plant Foods, Hohenheim University, D-70599
Stuttgart, 4Solvay Pharmaceuticals GmbH, D-30173 Hannover, 5Clinic of
6
Internal MedicineIII, Jena University, D-07743 Jena, Institute of Nutritional
Sciences, Giessen University, D-35392 Giessen
In a randomised cross-over design with 12 male and female volunteers the
antioxidant capacity as well as the content of phenolics and hippuric acid in
the 24 hours urinary excretion was assessed after ingestion of apple juice
(cultivar Bohnapfel) or a composite berry juice (composed of red grape,
blackberry, sour cherry, and blackcurrant juice) either. The juices were tested
freshly produced and after a 4 months storing period. The antioxiant activity
was measured by using the TEAC- and the PCL-assay, total phenolics with
the aid of Folin assay and hippuric acid by LC-MS analysis.
After intake of both juices in equimolar doses in terms of the antioxidant
activity, the TEAC- and PCL-values as well as the total phenolics increased
significantly in the urinary excretion compared to controls after water intake.
The simultaneously enhanced hippuric acid excretion in the test group
revealed that the bioavailable phenolic compounds were partly excreted as
metabolites. The excretion of phenolics and their metabolites of the apple
juice exceeded weakly those of the berry juice, though no differences
appeared in the bioactivity.
In comparison to freshly produced the ingestion of the stored juices led to a
lowered antioxidant activity and a reduced content of phenolics and hippuric
acid in the urinary excretion, indicating a diminished availability of phenolics
after storage.
The study showed that a remarkable part of phenolics from fruit juices is
bioavailable and the absorbed compounds and/or their metabolites are in
vivo bioactive in affecting the antioxidative potential of body fluids.
31
Health aspects of supplementation with a functional food based
on cereals and onion
Comparing antioxidant properties of fruit juices with in
and in vivo assays
Kitti Nemeth1 – Jana Kadrabova2 - Alexander Madaric2 - Igo Kajaba2 - Anna
2
2
2
2
Nagyova – Csilla Mislanova –Zuzana Hudecova – Martina Bacekova
1
Food Research Institute, Priemyselna 4, 824 75 Bratislava, Slovakia
2
Slovak Medical University, Department of bioactive compounds and
nutritional screening, Limbova 12, 833 03 Bratislava 37, Slovakia
I.Bitsch, M. Netzel, G. Netzel, E. Carlé, R. Bitsch
In a randomised intervention study, volunteers were offerred cereal onion
biscuit along with green tea (quercetin intake of 49.3 mg/day) for two months.
From the initial 36 volunteers, 32 accomplished the study (aged 62 ± 7
years). Blood samples were analysed for biochemical, antioxidant and
immunological parameters before and 2 months after the supplementation as
well as after a two-month washout period.
Preliminary analysis of biochemical parameters points to a beneficial effect of
the supplementation on lipid metabolism resulting in decreasing trend in
atherogenic index. Blood samples were assayed for 16 biochemical and 9
heametological parameters.
Changes in antioxidant enzymes were as follows: glutathion peroxidase
activity decreased slightly but significantly (p<0.04), that of superoxide
dismutase did not change significantly, and the levels of endogeneous
antioxidant glutathion increased significantly in both plasma and red blood
cells after combined cereal biscuit and green tea consumption.
Supplementation did not affect total antioxidant capacity, whereas FRAP
values decreased. This decrease correlated with the lowered plasma uric
acid level. The supplementation significantly decreased malondialdehyde
concentration, the terminal product of lipid oxidation in plasma, and this fact
points to a protective antioxidant activity. Increased level of homocysteine is
an independent risk factor for the development of cardiovascular diseases.
After intervention, the levels of this parameter dropped significantly (by 18%,
p<0.000) whereas levels of methionine, which plays a role in homocysteine
metabolism, increased (by 12%, p<0.000). The levels of vitamin B12 and
folic acid, which are also important for keeping optimum homocysteine level,
increased, in case of the former significantly (p<0.008). Supplementation with
cereal onion biscuits and green tea resulted in statistically highly significant
increase of quercetin level in blood, 6 times the basal level on average.
Conclusions
The cereal biscuit supplementation was most efficient in decreasing
malondialdehyde levels, which is one of the factors of oxidative stress, and
increasing endogeneous glutathion. The fact that the supplementation
decreased MDA level may point to antioxidative effect of the supplemented
diet.
Another interesting result was the decrease of homocysteine and increase in
vitamin B12 level in blood and this may positively affect the risk of
cardiovascular disease development.
32
vitro
Introduction - Several fruit juices are rich in polyphenols with high
antioxidative activity and it has been demonstrated that after juice
consumption some unchanged compounds and several of their
conjugates and metabolites are present in human plasma and are
excreted in the urine (Netzel et al. 2002, Bitsch R. et al. 2004 Bitsch
I. et al 2004).
Because of rather poor bioavailability and intensive metabolism of
plant polyphenols in the human body there are great chemical
differences between the complex of antioxidant compounds in the
juices and that in the plasma of humans after consuming those
drinks. That means that plasma and tissues of the human body are
not exposed to the original antioxidant compounds but to a lot of
derivatives. Up to now it is largely unknown to what extent the in vitro
tests of bioactivity reflect the conditions in vivo. Therefore we tested
the question if there are correlations between the antioxidant capacity
of juices from apples, elderberry and blackcurrant and the resulting
increase in the antioxidant capacity of plasma after drinking them.
Study design - After an overnight fasting for 12 hours 4 females and
2 males (aged 24-34 years) ingested a single portion of apple juice
(700 ml) or elderberry juice (400 ml) or blackcurrant juice (200 ml) in
a 28-day interval. Plasma samples were collected in EDTA coated
tubes before (baseline) and over a period of 4 hours after juice
consumption. The plasmatic antioxidant capacity was assessed by
using the Trolox Equivalent Antioxidant Capacity (TEAC)-test
according to Miller et al (1993) with slight modifications, measuring
the prolongation of the lag-phases. Pharmacokinetic parameters
were determined from concentration-time curves of the plasma
TEAC-test, using a two-compartment open model which was best
fitted with B-values between 0.955 – 0.997 (apple juices) and 0.935 –
0.951 ( red coloured juices). The corresponding areas under the
curves (AUC), the mean residence time (MRT), the maximum
measured concentration Cmax and the corresponding time point
tmax were calculated with the pharmacokinetic programm TOPFIT
2.0.
Conclusion - Considering the areas under the plasma TEAC
concentration-time curves and the area-to-dose ratios after juice
consumption it becomes evident, that drinking “Brettacher” apple
juice is most suitable concerning the bioavailability of its antioxidative
compounds, particularly in contrast to blackcurrant juice.
These results demonstrate, that the simple in vitro estimation of the
antioxidative capacity of fruit juices and possible other foods does not
reflect the complex interactions in the organism which affect
bioavailability and metabolism of their antioxidative compounds. The
most significant method for the determination of their bioactivity
therefore remains the human study.
37
Analysis of Some Phytochemicals from Brussels sprouts and
Spinach
Bioaccesibility of carotenoids and tocopherols from vegetables:
Effect of minimal processing (modified atmospheres)
Mirjana Andjelkovic, Carmen Socaciu, Andrea Bunea, Madalina Neacşu,
Otilia Bobis, Roland Verhe, John Van Camp; 1) Department of Food Safety
and Food Quality, Ghent University, B-9000 Ghent, Belgium; 2) Department
3)
of Organic Chemistry, Ghent University, B-9000 Ghent, Belgium;
Department of Biochemistry, University of Agricultural Sciences and
Veterinary medicine, RO-400372 Cluj-Napoca, Romania
Olmedilla, B; Granado, F; Herrero, C; Pérez-Sacristán, B; Blanco, I;
Blázquez, S., Unidad de Vitaminas. Hospital Universitario Puerta de Hierro.
28035-Madrid. [email protected]
It is well known that vegetables are very rich sources of phytochemicals with
antioxidant capacity. Carotenoids and phenolic acids are an important class
of antioxidant components, of which few data are reported about their
composition in Brussels sporuts (Brassica oleracea) and in spinach (Spinacia
olearace). In addition, the influence of processing and post-harvest storage
on these components needs further investigation.
Several factors may alter the bioavailability of bioactive compounds which
can be associated with the food matrix as well as with the subjects.
Bioaccesibility usually refers to the proportion of a food component available
for absorption and the use of “in vitro” models allow to study individually both
digestive stages and food-related factors affecting bioavailability.
Our study focused on the optimization of carotenoid and phenolic acid
separations from Brussels sprouts and spinach. Qualitative and quantitative
composition of these vegetables in phenolic acids and carotenoids was
assessed with HPLC and LC-MS. The composition was determined during
storage and before and after processing, namely blanching, boiling, and
microwave treatment.
To evaluate cualitative and quantitative changes in carotenoids and
tocopherols contained in fruits and vegetables and to assess the influence of
minimal processing on the final bioaccesibility of these phytochemicals, fresh
and minimally processed (modified atmospheres*) orange and brocoli (plus
cooking) were assayed by using an “in vitro” model (1) and adapted to
evaluate isomerization, hydrolisis and estability of carotenoids and
tocopherols from foods. Homogeneizacion, salivar, gastric and duodenal
phases, and incorporation into aqueous-micellar phase were studied and
carotenoids and tocopherols assayed by HPLC in crude and saponified
extracts (2, 3).
Results showed the presence of several phenolic acids in Brussels sprouts
and spinach. All of them are from the hydroxycinnamic group of phenolic
acid: caffeic acid, p-coumaric acid, ferulic acid, and sinapic acid, in Brussels
sprouts, compared to p-coumaric acid, o-coumaric acid, and ferulic acid in
spinach. The processing of vegetables imposed a decrease of phenolic acids
in Brussels sprouts as well as in spinach. Moreover the storage conditions at
4°C and time of storage contributed to the 15-25% reduction of phenolic
acids in spinach. In addition to the individual quantification by HPLC, the total
phenol content was calculated by means of the colorimetric method of FolinCiocalteu.
Hydrolisis of lutein, zeaxanthin and b-cryptoxanthin present in orange is
incompletely and to a similar extent regardless the use of minimallyprocessed techniques. The amount of free lutein, zeaxanthin and bcryptoxanthin ready for absorption do not change after 9 days of storing
minimally-processed vegetables at 4º C. Isomerization of lutein and bcarotene present in (cooked) broccoli and estability of lutein, b-caroteno, gand a-tocopherols did not change because of the minimally processing nor
after 9 days of storage. In conclusion, compared with non-processed foods,
bioaccesibility of major carotenoids and tocopherols is not altered in
minimally-processed orange and broccloli.
Concerning carotenoids, alterations of carotenoid profile due to the storage
and processing of these vegetables were observed, mainly due to the
decrease of neoxanthin, violaxanthin, β-carotene, and lutein.
1.- Oomen et al, (2003). Arch. Envirom. Contam. Toxicol., 44: 281-287.
2.- Granado et al (1992). J. Agric. Food Chem 40, 2135-2140, 1992.
3.- Olmedilla et al, (1997). Clin. Chem., 43 (6): 1066-1071.
* Prepared at Instituto del Frío (CSIC), Madrid, Spain
Keywords: Carotenoids, tocopherols, bioaccesibility, minimally processed.
36
33
Evaluation of Phenolic Profile Of Olive Oils
Countries
Abstracts
From European
M. Andjelkovic1,2, J. Van Camp2, V. Van Hoed1, R. Verhé1; 1Department of
Organic Chemistry, Faculty of Bioscience Engineering, University of Ghent;
2
Department of Food Safety and Food Quality, Faculty of Bioscience
Engineering, University of Ghent
The main objective of this work was to obtain a chemical identification of the
polyphenols present in different olive oils of monovarietal olive cultivars and
to associate their presence with the determining factors such as cultivar,
climate of the production region and olive oil production steps.
Phenolics from olive oils from France, Italy, Portugal, Spain, and some other
Mediterranean countries were extracted using solid phase extraction and
afterwards analyzed by LC-MS which was developed and validated within
this study. All analyses were done in triplicates. Subsequently, principal
component analysis, PCA was carried out to characterize the oil and
evaluate the possible correlation among the origin of olive oil and phenolic
compounds present. Additionally, determination and quantification of
tocopherols and sterols in these oils was done. Oxidative stability of oils was
assessed with the Rancimat method.
Poster Presentations
In alphabetical order (1st author)
In all analyzed samples hydroxytyrosol and tyrosol were major components,
while luteolin and apigenin were the main flavonoids. Beside these some
other components were identified: vanillic acid, p-coumaric acid, elenolic
acid, dialdehydic form of decarboxymethyl oleuropein aglycon, dialdehydic
form of decarboxymethyl ligstroside aglycon, aldehydic form of oleuropein
aglycon, aldehydic form of ligstroside aglycon, ligstroside aglycon.
Tyrosol is present in higher amount than hydroxytyrosol, and the other
compounds are present in noticeable lower amounts. Samples collected from
Italian localities contained the highest amount of phenols compared to other
olive oils. PCA gave an overall observation about the correlation of 70 oil
samples according to their phenolic profile. The variance is explained in the
first two principal components.
Different phenolic profiles of olive oils from different regions were
established. PCA has shown to be of significant value for the correlation of
cultivar, origin, and some other factors.
34
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program

Transcription

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