Você está na página 1de 13

Journal of the Science of Food and Agriculture

J Sci Food Agric 80:10811093 (2000)

Review
Flavonols, flavones and flavanols nature,
occurrence and dietary burden
Peter CH Hollman1* and Ilja CW Arts1,2
1

State Institute for Quality Control of Agricultural Products (RIKILT), Bornsesteeg 45, NL-6708 PD Wageningen, The Netherlands
National Institute of Public Health and the Environment, PO Box 1, NL-3720 BA Bilthoven, The Netherlands

Abstract: Total avonol and avone contents of foods have been determined with validated state-ofthe-art methods. Quercetin dominates, and avonol levels found in vegetables and fruits are below
10 mg kg1. However, high concentrations are found in onions (300 mg kg1), kale (450 mg kg1),
broccoli (100 mg kg1), beans (50 mg kg1), apples (50 mg kg1), blackcurrants (40 mg kg1), and tea
(30 mg l1). The dietary intake of avonols varies 10-fold between countries (660 mg day1). Flavones
are of minor importance in the diet. Tea, wine and fruits are the most important sources of avanols,
but there are gaps in our knowledge on avanol levels of many foods. The absorption of dietary
quercetin glycosides in humans ranges from 20 to 50%. The sugar moiety is an important determinant
of the bioavailability of avonols. The presence of a glucose moiety signicantly enhances absorption.
The extent of absorption of avanols in humans seems similar to that of avonols but has been little
studied. Flavonols and avanols are extensively metabolised, as only 12% of them are excreted with an
intact avonoid backbone. Hepatic biotransformations include glucuronidation and sulphatation of
the phenolic hydroxyls and O-methylation of catechol groups. Bacteria of the colon cleave the C-ring of
the avonoid nucleus to phenolic acids which are subsequently absorbed. Apart from conjugates,
virtually no metabolites have been characterised in humans. Absorption of avanols is rather fast, with
times to reach peak values between 0.5 and 4 h. Flavanols are rapidly excreted, with elimination halflives of 16 h. Quercetin glycosides show rapid to slow absorption; peak values are reached between
< 0.5 and 9 h. The type of glycoside determines the rate of absorption. Excretion of quercetin glycosides
is slow: elimination half-lives are 24 h, independent of the type of glycoside. Analytical data for
avanols in foods are needed. Tea, as an important dietary source, has to be studied. Research on the
bioavailability of avonols and avanols has to be expanded. Attention is needed for the identication
and quantication of their metabolites in body uids.
# 2000 Society of Chemical Industry

Keywords: avonols; avones; avanols; catechins; glycosides; food contents; dietary intake; bioavailability;
absorption metabolism; phenolic acids

INTRODUCTION

Flavonols, avones and avanols or catechins constitute three of the major subclasses (Fig 1) of avonoids.
Flavonols and avones have a similar C-ring structure
with a double bond at the 23 position. Flavones, as
opposed to avonols, lack a hydroxyl group at the 3position. Major avonols are quercetin (3,5,7,3',4'pentahydroxyavone), kaempferol (3,5,7,4'-tetrahydroxyavone), myricetin (3,5,7,3',4',5'-hexahydroxyavone) and isorhamnetin (3,5,7,4'-tetrahydroxy-3'methoxyavone). The most abundant avones in
plants are luteolin (5,7,3',4'-tetrahydroxyavone) and
apigenin (5,7,4'-trihydroxyavone).
The discriminating structural feature of avanols,
which they have in common with anthocyanidins only,
is the lack of an oxygen group at the 4-position of the

heterocyclic C-ring (Fig 1). The lack of a double bond


at the 23 position and the presence of a 3-hydroxyl
group create two centres of asymmetry (carbons at
positions 2 and 3). So far, only avanols with a 2R
conguration have been found in nature. The predominating avanols are ()-catechin (2R,3S3,5,7,3',4'-pentahydroxyavan),
()-epicatechin
(2R,3R-3,5,7,3',4'-pentahydroxyavan), ()-gallocatechin (GC) (2R,3S-3,5,7,3',4',5'-hexahydroxyavan)
and
()-epigallocatechin
(EGC)
(2R,3R3,5,7,3',4',5'-hexahydroxyavan) and the following
gallic acid esters: ()-epicatechin gallate (ECg) and
()-epigallocatechin gallate (EGCg) (Fig 2). In the
past there has been confusion regarding nomenclature:
avanols are also referred to as avan-3-ols or
catechins. Strictly, the monomers should not be

* Correspondence to: Peter CH Hollman, State Institute for Quality Control of Agricultural Products (RIKILT), Bornsesteeg 45, NL-6708 PD
Wageningen, The Netherlands
(Received 29 October 1999; accepted 10 November 1999)

# 2000 Society of Chemical Industry. J Sci Food Agric 00225142/2000/$17.50

1081

PCH Hollman, ICW Arts

Figure 1. Subclasses of flavonoids. Classification is based on variations in


the heterocyclic C-ring.

described as proanthocyanidins or tannins. The latter


is an ancient name, referring to a complex mixture of
compounds that was used to bind proteins in the
tanning of animal hides. Both proanthocyanidins and
tannins include oligo- and polymeric forms of the
monomeric avanols. Polymerisation of monomeric
avanols can occur as a result of autoxidation, but
more often it is catalysed by polyphenoloxidase (PPO),
an enzyme which is present in most plant tissues.1 In
intact tissues, polyphenoloxidase is separated from its

phenolic substrates. Upon crushing or fermentation,


as in the preparation of tea and cacao, polymerisation
of avanols gives the characteristic brown pigments.
Oxidation processes which are induced to produce
black tea reduce its avanol content substantially. The
nature of these transformation products is discussed in
Ref 2.
Flavonols and avones are usually found in plants
bound to sugars as O-glycosides. Flavones may also
occur as C-glycosides. The glycosidic form is a general
feature of avonoids, with the exception of avanols
where glycosides are rare. Free avonoids, ie avonoids without their attached sugars, are called
aglycones. Aglycones of avonols and avones are
not present in fresh plants but may be present as a
result of food processing. Sugars are predominantly
bound to the avonoid nucleus via a b-glycosidic
bond. Sugar molecules can bind to various positions in
the parent avonoid, although there is a preference for
the 3-position. More than 80 different sugars have
been found bound to avonoids in plants. Monosaccharides (10 species), disaccharides (39), trisaccharides (30) and even tetrasaccharides (1) occur in
these glycosides. As a result, 179 different glycosides of
quercetin alone have been described in nature.3 A
multitude of glycosides also occur in common foods
such as tea4 and apples.5,6

OCCURRENCE

Early interest in avonoids focused on their unequivocal identication to study evolutionary and taxonomic relationships.7 Thin layer chromatography
(TLC) methods with spectrophotometric measurements were applied.8 However, in order to be able to
evaluate the biological effects of avonoids, reliable
data on avonoid contents of common vegetables and
fruits are needed. Over the last 20 years analytical
methods have evolved and nowadays selective and
sensitive HPLC methods are used.9 Tables 1 and 2
summarise the contents of avonols, avones and
avanols of edible parts of foods. For avonols and
avones only data obtained by HPLC methods with
conrmed peak identity (photodiode array detection
or LCMS) have been selected.
Total flavonols and flavones

Figure 2. Structures of flavanols: ()-catechin (I), ()-epicatechin (II, R 


H), ()-epigallocatechin (II, R  OH), ()-epigallocatechin (II, R  OH),
()-epicatechin-3-gallate (III, R  H) and ()-epigallocatechin-3-gallate (III
R  OH).

1082

The total avonol and avone contents of fresh and


processed vegetables, fruits and beverages commonly
consumed in the Netherlands have been determined.10,11 Fresh foods were purchased in a supermarket, a grocery and a street market. Brands of
processed foods were selected based on market share.
In addition, seasonal variation in contents was
determined. Flavonols and avones were determined
after hydrolysis of their glycosides, and peak identity
and purity were conrmed with photodiode array
detection.12 Crozier et al 13 and Justesen et al 14
followed the same hydrolysis procedure. In addition,
on-line LCMS was applied for peak identication.14
J Sci Food Agric 80:10811093 (2000)

Flavonols, avones and avanols

Table 1. Content of flavonols and flavones


(mg kg1 fresh weight or mg l1) in foods
determined with HPLC methods after hydrolysis of
their glycosides. Only edible portions are
considered

Food

Quercetin

Kaempferol

Myricetin

Luteolin

Apigenin

Apple
Apple juice
Apricot
Bean, French
French processed
Green
Slicing
Broad
Broad processed
Broccoli
Brussels sprouts
Cabbage, green
Red
Red processed
White
Cauliower
Celery, leaf
Stalk
Cherry
Processed
Cucumber
Currant, black
Red
Endive
Grape, black
White
Grape juice
Grapefruit juice (fresh)
Kale
Leek
Lettuce
Mushroom
Onion
Orange juice
Pea
Processed
Peach
Pear
Plum
Spinach
Strawberry
Sweet pepper, red
Tea, black
Tomato
Cherry tomato
Tomato juice
Wine, red

2036
2.5
2526
39
17
16
29
20
5.5
3037
06

4.6
2.8

1015

37
813
< 1.3
1537
212
4.4
4.9
110120

1479

340347
5.7

3.4
915

68.6

1417
214
63
13
8.3

< 12
< 3.8

<7
6072
7.49

46

211470
3031

512

1416

26

4.5
4.5
6.2

3.0

7.9

200
520

511

750
1661

, below limit of detection


Source: Hertog et al,10,11 Crozier et al 13 and Justesen et al. 14 Average values reported in these papers
are given. Whenever average values of two or more papers were available, the lowest and highest
averages are given.

Quercetin levels in vegetables were generally below


10 mg kg1, except for onions (280490 mg kg1), kale
(110 mg kg1), broccoli (30 mg kg1) and beans (45
60 mg kg1) (Table 1). Kaempferol could only be
detected in kale (210470 mg kg1), endive (15
90 mg kg1), broccoli (60 mg kg1) and leek (10
60 mg kg1). In most fruits the quercetin content
averaged 15 mg kg1, except for apples (20
70 mg kg1), apricot (25 mg kg1) and blackcurrants
J Sci Food Agric 80:10811093 (2000)

(37 mg kg1). The content of myricetin, luteolin and


apigenin generally was below the limit of detection
(about 1 mg kg1), except for beans (30 mg kg1
myricetin), sweet red pepper (1530 mg kg1 luteolin)
and celery stalks (520 mg kg1 luteolin, 15
60 mg kg1 apigenin). Quercetin levels in red wine
were between 5 and 15 mg l1, whereas in fruit juices
they were generally below 5 mg l1. Various black tea
infusions (n = 17) contained 1025 mg l1 quercetin,
1083

PCH Hollman, ICW Arts


Table 2. Content of flavanols (mg kg1 fresh weight or mg l1) in foods determined with HPLC methods. Only edible portions are considered

Food

()-Catechin

()-Epicatechin

Remarks

Reference

Apple
Apple without skin
Apple/peach juice
Apple juice
Apricot
Beer
Grape juice
Peach
Peach/grape juice
Peach nectar
Pear
Plum
Prune juice
Red grapes
Sour cherry
Sweet cherry
Tea, black

Trace17
Trace55
3
Trace2.3
2657
5
Trace2.03
50129
Trace3
Trace4.64
ND10
536
841
< 10
ND15
512
No data available

2101
10140
4
ND0.74
67171
No data available
Trace1
315
1
Trace3.60
159
016
ND
1821
4152
1449
3179

31
58
48
48, 63
31
68
48
31
48
48
31, 49
31
47
46
31
31
38

Tea, green

No data available

1094

Wine, red
Wine, white
Wine, port

ND208
716
115

1588
No data available
Trace10

16 varieties
8 varieties
Commercial juices
Commercial juices and concentrated juices
3 varieties
1 type
Commercial juices
4 varieties
Commercial juices
Commercial juices
Over 10 varieties
8 varieties
Various processing methods
Without seeds, 2 varieties
6 varieties
5 varieties
EGC: < 591
EGCg: 18229
ECg: 8110
EGC: 20287
EGCg: 60408
Over 800 types
2 types
7 types

37, 38
30, 68, 113
68
114

ND, below limit of detection.


EGC, ()-epigallocatechin; EGCg, ()-epigallocatechin gallate; ECg, ()-epicatechin gallate.

717 mg l1 kaempferol and 25 mg l1 myricetin. No


luteolin or apigenin was detected in any of the
beverages
Seasonal variation was large in leafy vegetables such
as lettuce (230 mg kg1 quercetin), endive (15
95 mg kg1 kaempferol) and leek (1156 mg kg1
kaempferol). Flavonol contents of these vegetables
were three to ve times higher in summer than in other
seasons.11 As the synthesis of avonoids in plants is
light-dependent, this is to be expected.
Crozier et al 13 studied the effect of cooking on the
quercetin content of onions and tomatoes. With both
foods, boiling reduced the quercetin content by 80%,
microwave cooking by 65% and frying by 30%.
Only limited data on the effect of processing are
available. In general, avonol levels in processed foods
were lower than in fresh products.11 However, varietal
differences could have played a role. Flavonol contents
of green and black tea were similar.10
Citrus peel and peel oils contain a series of
polymethylated avones that occur as aglycones.
Although concentrations may reach some 1.9
6.5 g l1 in peel oil,15 dietary burden is very low, since
levels in citrus juices only reach some 7 mg l1.16
Flavonol and flavone glycosides

Herrman17 reviewed the occurrence of avonol and


avone glycosides in vegetables. At that time only
qualitative data were available. The preferred bonding
site of the sugar is the 3-position, less frequently the 7position, and only in rare cases the 4'-, 3'- and 51084

positions. Flavones occur mainly as 7-O-glycososides,


but C-glycosides (the sugar is directly attached to an
aromatic carbon atom) are also known. Comparatively
few data are available for the avone-C-glycosides,
although they occur in teas18,19 and cereals.2022 DGlucose is the most frequent sugar; other sugar
residues found are D-galactose, L-rhamnose, L-arabinose, D-xylose and D-apiose, but also D-glucuronic
acid. In general, the sugars of the D-conguration
occur as b-glycosides, whereas those of the L-conguration occur as a-glycosides. In onions the major
glycosides are quercetin-4'-glucoside and quercetin3,4'-diglucoside.23,24 Green beans contain mainly
quercetin-3-O-glucuronide (414 mg kg1).25 Commercial processing did not result in chemical breakdown of the conjugates. The two main glycosides of
broccoli
were
quercetin-3-O-sophoroside
and
kaempferol-3-O-sophoroside
(65 mg kg1)
(166 mg kg1).26
Dick et al 5 identied the following glycosides of
quercetin in apple peel: 3-O-a-L-arabinoside, 3-O-b-Dgalactoside, 3-O-b-D-glucoside, a-L-rhamnoside and
3-O-b-D-xyloside. They used HPLC after acid hydrolysis as a separation tool for the aglycones. The identity
of the anomeric conguration of the glycosides was
determined with NMR. The extraction procedure
used was not suitable for quercetin di- or oligosaccharides. All sugars were in the pyranose conguration,
except the arabinoside which was a furanose. Lister et
al 6 compared two apple cultivars and found a similar
composition and concentration of avonol glycosides
J Sci Food Agric 80:10811093 (2000)

Flavonols, avones and avanols

in their peels. In addition to the aforementioned


glycosides, they identied quercetin-3-O-arabinoside
in the pyranose form, and quercetin-3-O-rhamnosylglucoside (rutin), and conrmed that the rhamnoside
is also attached to the 3-position. The peel contained
about 900 mg g1 quercetin glycosides, of which the
galactoside, the rhamnoside and the aribinoside
(furanoside) each contributed about 25%, the xyloside
15% and the glucoside 10%. Glycosides decreased
during ripening.
Price et al 27 determined avonol glycosides in tea
infusions brewed from commercially available black
teas in the UK. On average, the dominant glycosides
were quercetin-3-O-rhamnosylglucoside (25%) and
quercetin-3-O-glucoside (25%), followed by quercetin-3-O-galactoside (20%) and quercetin-3-O-glucosylrhamnosylglucoside
(20%).
The
dominant
kaempferol glycoside was kaempferol-3-O-glucoside
(33%), followed by kaempferol-3-O-rhamnosylglucoside (25%) and kaempferol-3-O-glucosylrhamnosylglucoside (25%). However, the avonol glycoside
prole varied in the different tea varieties.
Flavanols

Flavanols are widespread in plant foods; they have


been reported in tea,2830 fruits31,32 and legumes.33,34
Many of the older data on avanol levels in foods were
obtained with spectrophotometric detection methods
using vanillinHCl or FolinCiocalteu reagents.9
These reagents also react with components other than
avanols and are therefore unreliable if not combined
with proper separation methods.35,36 HPLC is the
method of choice, but for completeness, data obtained
with spectrophotometric measurements combined
with thin layer chromotography (TLC) will be
reported whenever no other data are available. Table
2 summarises avanol contents measured with HPLC.
Tea is probably the most important avanol source
in many countries. It combines a high level of
consumption with a relatively high avanol content.
Furthermore, it is the only plant food for human
consumption found so far which contains ()-epigallocatechin gallate (EGCg). The most abundant
monomeric avanols of black tea are ()-epicatechin
gallate (ECg), ()-epigallocatechin (EGC) and
EGCg. Only a small part of the avanol content of
teas is constituted by ()-catechin, ()-epicatechin
and ()-gallocatechin.28 Flavanols are sometimes
conveniently called tea avonoids, and tea has been
tested extensively for its biological actions in in vitro
and in vivo animal experiments. However, surprisingly
few data on avanol contents of tea brews are
available. Studies on the chemistry of tea typically
report avanol contents of fresh tea leaves on a dry
weight basis after exhaustive extraction with organic
solvents. Such data do not take into account infusion
rates into ordinary hot water and variations in brewing
methods among countries. Table 2 presents data from
two studies where tea brews were analysed.37,38 More
data for a better interpretation of epidemiological
J Sci Food Agric 80:10811093 (2000)

studies on the health effects of tea consumption are


needed.
Relatively high levels of ()-catechin and ()-epicatechin have been reported in red wine, and much
lower levels in white wine.3942 Comparison of data
reported in the literature is difcult, since grape
cultivar has such a pronounced impact on the avanol
concentration of a certain wine. Goldberg et al 30
reported data on the avanol content of 836 red wines
from a large number of wine-making regions of the
world. They noted the exceptionally high avanol
content of wine made from Pinot Noir grapes (up to
287 mg l1). Most of the wine from Burgundy, France
is made from this grape. Next to grape cultivar, which
is the most important determinant of the avanol
content of wine, climatic conditions appear to play an
important role. Within cultivars the highest avanol
levels were found in wines grown under damp cool
conditions, whereas dry sunny climates yielded lower
avanol concentrations.30 Even within France an
association could be observed between the average
temperature in a region and the avanol content of the
wine produced in that region.39 However, winemaking methods differ between regions as well and
may explain some of the observed variation attributed
to climate.
In contrast with the abundance of data on avanols
in wine, data on grapes are limited. Qualitative studies
show the presence of ()-catechin, ()-epicatechin
and ECg in black and white grape seeds and skins.4345
However, ECg was not present in the skins of all
varieties that were tested.44 The one quantitative study
on avanols in black grapes reports low levels.46 The
authors admit that these levels are extremely low, even
compared with white grapes grown in the same region.
Flavanols have been determined in apricot, pear,
cherry, peach and plum (Table 1).31,4750 Herrmann
and co-workers33,51,52 reported the presence of avanols in strawberries, black, white and red currants,
gooseberries, blueberries and rhubarb. These authors
used colorimetric detection after TLC separation. Van
Gorsel et al 47 could not conrm the TLC data on
()-epicatechin in juice made from strawberries.
Rommel and Wrolstad53 detected ()-catechin and
()-epicatechin in raspberry. Low levels of ()-epicatechin were reported in avocado.54 Only qualitative
data are available for legumes34,55 and cacao and
chocolate.56,57 These foods are probably high in
avanols and should be examined. Part of the variation
in reported avanol contents may have been caused by
variations in the developmental stage of the fruits. For
apples and pears it has been shown that during growth
there was a rapid decrease in avanol levels. During
maturation the uctuations were low.49,58 However,
information about developmental changes in fruits
other than apples and pears is not available. Storage of
apples, harvested at commercial maturity, under
normal cold storage conditions for over 6 months gave
only very small changes in ()-catechin and ()-epicatechin content.58,59
1085

PCH Hollman, ICW Arts

Crushing and fermentation are major processing


steps in the preparation of tea and cacao. As a result,
oxidation and polymerisation of avanols can reduce
the content of monomeric avanols by as much as
90%.60 The avanol content of black tea is much lower
than that of green tea, where oxidation is almost
completely prevented.28 Processing is not an important issue for avanols in fruits, since they are usually
consumed raw. However, in fruit juices, processing
may seriously affect avanol contents. Home-made
juice from apples used for commercial apple juice
manufacture61 contained the expected levels of ()catechin and ()-epicatechin. Surprisingly, only traces
or very low concentrations of avanols were found in
commercial apple juice (up to 2.3 mg l1 ()-catechin
and up to 0.7 mg l1 ()-epicatechin.48,6264 Spanos et
al 62 showed that the complex process of commercial
apple juice preparation decreased its avanol content
in a stepwise manner. In particular, crushing and
pressing, storage of the concentrated juice at room
temperature and decolorisation by treatment with
activated carbon destroyed the avanols entirely. The
same applies to grape juice: relatively high levels in
home-made juice,65 whereas commercial juice, depending on the manufacturing method used, does not
contain any avanols.66 Low levels of ()-catechin
(5 mg l1) and ()-epicatechin (1 mg l1) have
been reported in lager beers.6769 Polyvinylpolypyrrolidone (PVPP) is an adsorbent for phenolics which is
used by breweries to prolong the stability of beers
against haze formation. PVPP was shown to reduce the
avanol content of lager beer.69,70
Seto et al 71 showed that heat treatment may induce
epimers of individual avanols. This could have
important implications for their pharmacodynamics.
If avanols exert their effect via modulation of enzyme
systems, structural epimers may not have the same
effects. It was shown that epimerisation hardly
proceeds at 80 C (pH 5), whereas at 100 C some
effect could be detected only after 30 min. Moreover,
epimerisation was very limited at pH < 5 (at 120 C).
To summarise, epimerisation of avanols only occurs
when a product is exposed to high temperatures at
pH > 5 for a prolonged period. These conditions are
exceptional in food processing; even tea is probably
not boiled for 30 min.
Dietary intake of flavonols, flavones and flavanols

The average intake of avonols and avones in the


Netherlands was 23 mg day1, of which the avonol
quercetin contributed 16 mg day1, kaempferol
3.9 mg day1 and myricetin 1.4 mg day1. Thus avones contributed only a minor fraction, about 7%.
Tea turned out to be the major source in this
population (48% of total intake), followed by onions
(29%) and apples (7%).72 This is in contrast with data
of Kuhnau,73 who estimated that the total intake of
avonoids in the USA was 1 g day1 (expressed as
glycosides). This would be equivalent to about
115 mg day1 of avonol and avone aglycones, as
1086

opposed to 23 mg day1 in our studies. Most likely the


avonoid contents used by Kuhnau73 were too high,
not only because they were obtained with methods
now considered obsolete, but also because sometimes
avonoid contents of non-edible parts were included.
Finally it can be argued that the food disappearance
values used by Kuhnau73 tend to overestimate food
intake.
The intakes of avonols in Japan, the Netherlands,
the former Yugoslavia, the United States, Finland,
Italy and Greece were determined by quantifying the
avonols in equivalent food composites representing
their average diet around 1960.74 The main sources of
avonols were tea in Japan and the Netherlands, red
wine in Italy and onions in the USA, the former
Yugoslavia and Greece. In Finland, berries are an
important source of avonols. Flavonol intake was
highest in Japan (64 mg day1) and lowest in Finland
(6 mg day1). Average intakes of avonols in a number
of epidemiological prospective cohort studies published so far are 20 mg day1 in middle-aged to older
American males,75 26 mg day1 in elderly Dutch
men,76 4 mg day1 in Finnish middle-aged men and
women77 and 26 mg day1 in Welsh middle-aged
men.78
Tea, wine and fruits are the most important sources
of avanols, but there are gaps in the knowledge on
avanol levels in many foods. Surprisingly, only very
limited data on avanol contents of tea brews are
available. As a result, it is at present not possible to
estimate the intake of avanols in a human population.

ABSORPTION, BIOAVAILABILITY AND


METABOLISM

Knowledge on the absorption, bioavailability and


metabolism of dietary avonoids is important to fully
evaluate their potential benecial role in human
health. We reviewed animal as well as human data
on these topics.79,80 This paper updates those reviews
for avonols, avones and avanols and focuses only
on human data.
Absorption and bioavailability of flavonols and
flavones

Absorption of avonols from the diet was long


considered to be negligible, because avonols are
present in plants bound to sugars as b-glycosides. Only
aglycones were considered absorbable, whereas glycosides were considered non-absorbable. Studies with
germ-free rats indeed showed that large amounts of
unchanged glycosides were excreted with faeces,
whereas only small amounts of glycosides were found
in faeces of rats with a normal microora.81 Apparently, enzymes capable of splitting these b-glycosidic
bonds are not secreted into the gut or present on the
intestinal wall. Bacteria in the colon are able to
hydrolyse avonoid glycosides,8284 but at the same
time they degrade the liberated avonoid aglycones. In
addition, the absorption capacity of the colon is far less
J Sci Food Agric 80:10811093 (2000)

Flavonols, avones and avanols

than that of the small intestine. As a result, only


marginal absorption of glycosides was expected.73
The absorption of quercetin from regular foods in
man was studied in ileostomy subjects who lack a
colon with the bacterial ora.85 These subjects were
chosen to circumvent the problem of microbial
degradation. Surprisingly, absorption of the quercetin
glucosides from onions (52%) was far better than that
of the pure aglycone (24%). Absorption of pure rutin
(quercetin-3-O-rhamnosylglucoside), a major glycoside in tea, was 17%. Thus glycosides can be absorbed
in man as such without prior hydrolysis by microorganisms.
Dietary avonol glycosides showed very rapid to
very slow absorption in man;8689 times to reach peak
concentrations (Tmax) were between < 0.5 and 9 h
(Table 3). When the absorption of quercetin from
onions, apples and rutin in healthy subjects was
compared, distinct differences in rates of absorption
were found.87,88 The bioavailability of quercetin
glucosides from onions was superior to that of various
quercetin glycosides from apples (30%) and of pure
quercetin rutinoside (30%). Onions contain mainly
quercetin-b-glucosides, whereas apples contain a
mixture of quercetin-b-galactosides and b-xylosides,
and quercetin is bound to a disaccharide in rutin.
These data suggested that the sugar moiety of
quercetin glycosides is an important determinant of
their absorption and bioavailability, but left open the
possibility of matrix effects of the foods. The important role of the sugar moiety in the absorption of
quercetin was also found in the aforementioned study
with ileostomy subjects.85 Subsequently we investigated whether it is indeed the sugar moiety that
determines quercetin absorption in humans, by

comparing the time course of the quercetin concentration in plasma after administration of pure quercetinb-glucoside or pure quercetin-b-rutinoside to healthy
human volunteers.89 The peak concentration of
quercetin (Cmax) in plasma was 20 times higher and
reached (Tmax) more than 10 times faster after intake
of the glucoside than after the rutinoside (Table 3).
The bioavailability of the rutinoside was only 20% of
that of the glucoside. These pharmacokinetic data
suggest that quercetin glucoside was absorbed from
the small intestine, whereas quercetin rutinoside was
absorbed from the colon after deglycosylation.
If indeed the glucoside is absorbed from the small
intestine, this implies that the intact quercetin glucoside has to pass across the endothelial membrane for
the following reasons. One reason is that these bglycosides are resistant to hydrolysis by HCl in the
stomach.12 Another reason is that b-glycosidases are
not secreted into the small intestine.73 A third reason is
that the broad-specicity b-glucosidases needed to
hydrolyse quercetin glucoside are not bound to the
brush border membrane.90 Therefore avonoids conjugated with glucose might be carried into the smallgut enterocyte via active transport, for instance via the
intestinal Na/glucose cotransporter. The presence of
quercetin-4'-glucoside in human plasma91 also suggests that the intact glucoside may be absorbed. Model
studies showed that naphthol glucosides were transported by the active Na/glucose transporter across
the intestinal wall of rats.92 So far, only two in vitro
studies on absorption mechanisms of quercetin glucosides have been published. In everted sacs of rat
jejunum, quercetin glucosides were capable of binding
to the Na/glucose cotransporter in a sodium-dependent way.93 However, no evidence for active transport

Table 3. Pharmacokinetic parameters of flavonols and flavones in humans

Subjects

Oral dose (mg)

Cmax (ng ml1)

Quercetin aglycone

4000

< 100

Flavonol glycosides
from Gingko biloba
extract
Quercetin glucosides
from onions

28140

22.5

2
9
5

64a
68a
102 (quercetin-4'-glucoside)

200*
225*
45**

2.9
0.7
1.3

17
28

98a

90

2.5

23

Quercetin-4'-glucoside

9
9
9

100a
94a
94a

90
54
1057

9.3
6.0
< 0.5

28
22

Luteolin

50

Luteolin and
luteolin
monoglucuronide
present

Food, avonol

Quercetin glycosides
from apples
Quercetin-3-rutinoside

Tmax(h) T1/2(h)

Method

Reference

Plasma, aglycone
uorimetric
Plasma, total
avonols HPLC

115

*Plasma, total
quercetin HPLC
**Plasma,
quercetin-4'-glucoside
Plasma, total
quercetin HPLC
Plasma, total
quercetin HPLC
Plasma, total
quercetin HPLC
Serum, HPLC

87

86

88, 91
88
88, 89
88
98

Expressed as aglycone.
, data not provided.

J Sci Food Agric 80:10811093 (2000)

1087

PCH Hollman, ICW Arts

of quercetin glucosides was found in human intestinal


epithelial Caco-2 cells.94 Thus evidence for a role of
this transporter is still inconclusive.
Although nutritionally of minor importance, it was
shown in an intervention study that the aglycone of
quercetin was absorbed in humans.95 Thirteen subjects consumed 1000 mg quercetin aglycone and
200 mg quercetin rutinoside for 28 days; 14 subjects
consumed a rice our placebo for these 28 days. The
concentration of quercetin in the fasting plasma of the
quercetin-supplemented group after 28 days was 23fold higher than that of the placebo group.
The elimination of quercetin from plasma was slow,
taking about 24 h (Table 3), which implies that
quercetin may accumulate in plasma throughout the
day with repeated dietary intake. Accumulation of
quercetin in plasma was demonstrated in a study
where volunteers drank eight cups of tea every 2 h for a
period of 3 days (Hollman PCH et al, unpublished). In
a study with volunteers who ingested a quercetin-rich

meal, it was shown that the quercetin concentration in


plasma returned to baseline after 20 h.96 This would
imply a much shorter elimination half-life than shown
in Table 3. However, in this study the period of the
quercetin-free diet prior to the supplementation was
much too short. Consequently, baseline values of
plasma quercetin were too high.
Reduced absorption of tea polyphenols was invoked
to explain why consumption of black tea raised the
plasma antioxidant capacity in volunteers while tea
with milk did not.97 In a study with nine volunteers it
was found that addition of milk to black tea did not
change the area under the curve of the plasma
concentrationtime curves of quercetin or kaempferol
(Hollman PCH et al, unpublished). Thus bioavailability of avonols was independent of the addition of
milk.
Only one preliminary study on absorption of
avones, namely luteolin, has been published, which
showed that luteolin was absorbed.98

Table 4. Pharmacokinetic parameters of flavanols in humans

Food, catechin Subjects

Cmax (ng ml1) Tmax (h)

Oral dose

()-Catechin

tablet

3000 mg

15000

()-Catechin

Granulate
suspension

500; 1000;
1500 mg

Green tea

3 g lyophilised

green tea (6 cups)

570;
1113;
2276
685

Green tea

12

3 g lyophilised
green tea solids

Black tea

12

Green tea

3 g lyophilised
black tea solids
1.2 g lyophilised
decaffeinated
green tea

Green tea

Green tea

Green tea

18

Black chocolate 8

 930 mg total
avanols

160

 49 mg total
49
avanols
EGCg 88 mg
46268
EGC 82 mg
82206
ECg 33 mg
<1
EC 32 mg
4880
300 ml green tea EGCg level in
2360
brew
tea not provided
(6 g tea, 3 min)
Extract
EGCg 225; 375; 300; 1970;
(Sunphenon
525 mg
2020
DCF-1)
EGC 7.5; 12.5;
10; 44;
7.5 mg
78
1.5; 3.0; 4.5 g
EGCg 110; 220; 120; 325;
lyophilised
330 mg
320
decaffeinated
EGC 100; 200;
150; 510;
green tea
300 mg
550
EC 38; 75;
55; 190;
110 mg
190
40; 80 g black
EC 82; 164 mg
103; 196
chocolate

12

T1/2 (h)

Method

1.3

Serum, photometric,
free catechin only
1.6; 1.4; 1.3; 1.1; Serum, HPLC, free
2.0
0.9 catechin only
1

2.3

4.8

2.2

6.9

4
1

1
0.5

1.5
1.5

1.6; 2.4;
2.7
1.4; 1.8;
1.3
1.4; 1.8;
1.8
2.0; 2.6

Reference
100
99

Colorimetric (DMCA)
total avanols, free
avanols only?, no
hydrolysis described
Colorimetric (DMCA)
total avanols, free
avanols only?, no
hydrolysis described

103

Plasma, HPLC
Coulochem
electrode array, after
enzymatic hydrolysis
Plasma, HPLC photodiode
array, after acid hydrolysis

106

Plasma, HPLC with


chemiluminesence, free
avanols only

5.5; 5.0; Plasma, HPLC


4.9 Coulochem electrode
2.7; 2.8; array, after enzymatic
2.5 hydrolysis
5.7; 3.4;
3.2
1.9; 2.3 Plasma, HPLC
uorescence,
after enzymatic
hydrolysis

102

104

105

101

107

, data not provided.

1088

J Sci Food Agric 80:10811093 (2000)

Flavonols, avones and avanols

Absorption and bioavailability of flavanols

After oral administration of radioactively labelled ()catechin to rodents, monkeys and man, about 50% of
the total administered radioactivity was excreted into
urine.80 This indicates that ()-catechin and its
microbial degradation products are well absorbed.
By the 1970s the pharmacokinetics of ()-catechin
as a pure compound had been studied in a limited
number of volunteers (Table 4).99101 Peak levels of
free ()-catechin in serum were reached within 2 h,
and the elimination from plasma was rather rapid with
half-lives of only 1 h (Table 4). However, the high ()catechin concentrations found by Giles and Gumma100 are probably caused by the use of a photometric
method without prior chromatographic separation.
The pharmacokinetics of EGCg, EGC, ECg and
()-epicatechin was determined in volunteers who
ingested green tea and black tea extracts, the principal
dietary sources of these avanols (Table 4).101106
EGCg showed rapid to slow absorption: the time to
reach a peak value was between 0.5 and 4 h. The peak
values obtained by Lee et al 106 and Yang et al 101 after a
dose of 100 mg EGCg were similar; however, Nakagawa and Miyazawa105 found extreme high concentrations after doses of 375 and 525 mg EGCg.
Surprisingly, Nakagawa and Miyazama105 determined
only free EGCg in plasma, whereas the other authors
used an enzymatic hydrolysis of the conjugates. No
explanation is apparent for this discrepancy. Only
Yang et al 101 determined the elimination half-life of
EGCg, which was about 5 h. Time to reach peak
values after EGC was between 1 and 2 h; the
elimination half-life was 2.53 h. The pharmacokinetics of ()-epicatechin was determined after ingestion of tea101,106 and chocolate.107 Times to reach
peak values of plasma ()-epicatechin were similar,
between 1.4 and 2.6 h. Elimination half-lives of
()-epicatechin from plasma were between 2 and 6 h.
In summary, absorption and elimination of EGCg,
EGC, ECg and ()-epicatechin are rather fast. The
present studies do not allow the pharmacokinetics of
these various avanols to be distinguished.
The potentially reduced absorption of tea avanols
from tea with milk was studied in volunteers. As for
avonols, no effect of the addition of milk to tea could
be measured.102,108
Metabolism of flavonols, flavones and flavanols

In the metabolism of avonoids, two compartments


are important. The rst consists of tissues in the body,
where biotransformation enzymes located mainly in
the liver act upon absorbed avonoids and their
absorbed colonic metabolites. The kidney and the
small intestine might also contain enzymes capable of
biotransformation of avonoids. Conjugation of the
polar hydroxyl groups with glucuronic acid, sulphate
or glycine has been reported for avonoids and their
colonic metabolites.79 In addition, O-methylation by
the enzyme catechol-O-methyltransferase plays an
important role in the inactivation of the catechol
J Sci Food Agric 80:10811093 (2000)

moiety, ie the two adjacent (ortho) aromatic hydroxyl


groups, of avonoids and their colonic metabolites.
Indirect evidence for the presence of quercetin
conjugates in humans was obtained by Manach et
al,96 who could not detect quercetin agycone in
plasma. However, the quercetin concentration rose
to submicromolar concentrations after b-glucuronidase/sulphatase hydrolysis of the plasma. The presence of a monoglucuronide of luteolin was
demonstrated in human serum.98 The methylated
derivative of quercetin, isorhamnetin, was found in the
plasma of three out of 10 subjects who consumed a
quercetin-rich diet.96 However, this does not prove
that methylation had occured, because isorhamnetin
was probably present in the diet.
Flavanols appear to be present in plasma as free
avanols and as sulphates and glucuronides. The
prole depends on the type of avanol. Lee et al 106
determined avanol conjugates in plasma after ingestion of green tea. EGCg was mainly present as a
sulphate conjugate (65%), followed by the free form
(20%) and the glucoronide (15%). EGC was mainly
present as glucuronide (60%), followed by sulphate
(30%); about 10% was present as unconjugated EGC
aglycone. Only conjugated ()-epicatechin was found
with two-thirds sulphate and one-third glucuronide.
The second metabolically active compartment is the
colon, where micro-organisms degrade unabsorbed
avonoids and avonoids that are absorbed and then
secreted with bile. Colonic bacteria produce glycosidases, glucuronidases and sulphatases that can strip
avonoid conjugates of their sugar moieties, glucuronic acids and sulphates.109 Human intestinal bacteria
are able to hydrolyse O-glycosides109 as well as Cglycosides.110 In addition, degradation by microorganisms involves splitting of the heterocyclic oxygen-containing ring. The subsequent degradation
products can evidently be absorbed, because they are
found in urine.79 These include a variety of phenyl
carboxylic acids which, depending on their hydroxylation pattern, are antioxidants themselves111 and thus
may contribute to the biological effects of dietary
avonoids.
The type of ring ssion depends on the type of
avonoid: three schemes have been described.79
Flavonols are degraded to phenylacetic acids and
phenylpropionic acids. Ring ssion of avanols produces valerolactones (a benzene ring with a side chain
of ve Carbon atoms) and phenylpropionic acids.
Flavones and avanones follow a scheme producing
phenylpropionic acids. These phenylcarboxylic acids
are subject to further bacterial degradation and to
enzymatic transformations in body tissues. As a result,
the phenylpropionic acids will be oxidised to benzoic
acids.
Flavonols are extensively metabolised in humans.
Only 0.11.4% of the ingested dietary quercetin was
excreted as unchanged quercetin or its conjugates in
urine.85,88,89,91 Aziz et al 91 determined individual 4'glucosides in onions and urine and found that the
1089

PCH Hollman, ICW Arts

excretion in urine as a proportion of intake was 17%


for isorhamnetin-4'-b-glucoside and 0.2% for quercetin-4'-b-glucoside.
Extensive metabolism of avanols in humans has
been reported.80 Only 0.22% of the administered
avanols were found in plasma.112

CONCLUSIONS

Total avonol and avone contents of foods have been


determined with validated state-of-the-art methods.
The dietary intake of avonols varies 10-fold between
countries (660 mg day1). Flavones are of minor
importance in the diet. Flavonols and avones only
occur as glycosides in foods, which seriously affects
their bioavailability. However, only limited data on
contents of individual glycosides are available. This
limitation in our knowledge hampers a thorough
understanding of the dietary impact of avonols in
the body. Tea, wine and fruits are the most important
sources of avanols, but owing to gaps in information
on avanol levels of many foods and the lack of data on
tea brews, it is at present not possible to estimate the
intake of avanols by a human population.
The absorption of dietary quercetin glycosides in
humans ranges from 20 to 50%. The sugar moiety is
an important determinant of the bioavailability of
avonols. Flavonol glycosides can be absorbed as
intact molecules, and the presence of a glucose moiety
signicantly enhances absorption. The extent of
absorption of dietary avanols in humans has been
little studied. Flavonols and avanols are metabolised
predominantly in the liver and colon. Hepatic biotransformations include glucuronidation and sulphatation of the phenolic hydroxyls and O-methylation of
catechol groups. Unabsorbed avonols and avanols
and their conjugates secreted with bile into the small
intestine are degraded by bacteria in the colon. These
bacteria hydrolyse conjugates and cleave the C-ring of
the avonoid nucleus to phenolic acids which are
subsequently absorbed. Metabolism is extensive, as
only a 12% of the avonols and avanols are excreted
with an intact avonoid backbone. However, apart
from conjugates, virtually no metabolites have been
characterised in humans.
Absorption of avanols is rather fast, with times to
reach peak values between 0.5 and 4 h. Flavanols are
rapidly excreted, with elemination half-lives of 16 h.
The present studies do not allow the pharmacokinetics
of individual avanols to be distinguished. Quercetin
glycosides show rapid to slow absorption; peak values
are reached between < 0.5 and 9 h. The type of
glycoside determines the rate of absorption. Excretion
of quercetin glycosides is slow: elimination half-lives
are 24 h, independent of the type of glycoside.

FUTURE RESEARCH REQUIREMENTS

More extensive analytical data for avanols in foods


are needed. Tea as an important dietary source for
1090

avanols in many countries has to be studied.


Emphasis is needed on variations in tea brewing habits
and the effects of tea variety.
Research on the bioavailability of avonols, avones
and avanols has to be expanded. Attention must be
given to the identication and quantication of their
metabolites in body uids and tissues. Sensitive and
selective analytical methods will have to be developed.
With these tools, additional human studies will have to
be carried out to fully understand the pharmacodynamics of avonols, avones and avanols.

REFERENCES
1 Mayer AM and Harel E, Polyphenol oxidases in plants.
Phytochemistry 18:193215 (1979).
2 Harbowy ME and Balentine DA, Tea chemistry. Crit Rev Plant
Sci 16:415480 (1997).
3 Williams CA and Harborne JB, Flavone and avonol glycosides,
in The Flavonoids: Advances in Research Since 1986, Ed by
Harborne JB. Chapman and Hall, London, pp 337385
(1994).
4 Finger A, Engelhardt UH and Wray V, Flavonol glycosides in
teakaempferol and quercetin rhamnodiglucosides. J Sci
Food Agric 55:313321 (1991).
5 Dick AJ, Redden PR, DeMarco AC, Lidster PD and Grindley
TB, Flavonoid glycosides of Spartan apple peel. J Agric Food
Chem 35:529531 (1987).
6 Lister CE, Lancaster JE, Sutton KH and Walker JRL,
Developmental changes in the concentration and composition of avonoids in skin of a red and a green apple cultivar. J
Sci Food Agric 64:155161 (1994).
7 Swain T, Flavonoids as chemotaxonomic markers in plants, in
Pigments in Plants, Ed by Czygan FC. Gustav Fischer,
Stuttgart, pp 224236 (1980).
8 Mabry TJ, Markham KR and Thomas MB, The Systematic
Identication of Flavonoids. Springer, Berlin (1970).
9 Robards K and Antolovich M, Analytical chemistry of fruit
bioavonoids: a review. Analyst 122:R11R34 (1997).
10 Hertog MGL, Hollman PCH and van de Putte B, Content of
potentially anticarcinogenic avonoids of tea infusions, wines,
and fruit juices. J Agric Food Chem 41:12421246 (1993).
11 Hertog MGL, Hollman PCH and Katan MB, Content of
potentially anticarcinogenic avonoids of 28 vegetables and 9
fruits commonly consumed in the Netherlands. J Agric Food
Chem 40:23792383 (1992).
12 Hertog MGL, Hollman PCH and Venema DP, Optimization
of a quantitative HPLC determination of potentially anticarcinogenic avonoids in vegetables and fruits. J Agric Food
Chem 40:15911598 (1992).
13 Crozier A, Lean MEJ, McDonald MS and Black C, Quantitative analysis of the avonoid content of commercial
tomatoes, onions, lettuce, and celery. J Agric Food Chem
45:590595 (1997).
14 Justesen U, Knuthsen P and Leth T, Quantitative analysis of
avonols, avones, and avanones in fruits, vegetables and
beverages by high-performance liquid chromatography with
photo-diode array and mass spectrometric detection. J
Chromatogr A 799:101110 (1998).
15 Gaydou EM, Bianchini JP and Ramananarivo RP, Orange and
mandarin oils differentiation using polymethoxylated avone
composition. J Agric Food Chem 35:525529 (1987).
16 Veldhuis MK, Swift LJ and Scott WC, Fully methoxylated
avones in Florida orange juices. J Agric Food Chem 18:590
592 (1970).
17 Herrmann K, On the occurrence of avonol and avone
glycosides in vegetables. Z Lebensm Untersuch Forsch 186:1
5 (1988).
18 Kuhr S, Herzig B and Engelhardt UH, Studies of polymer

J Sci Food Agric 80:10811093 (2000)

Flavonols, avones and avanols

19

20
21
22

23
24

25

26

27

28
29

30

31

32
33

34

35
36

37

38

39

polyphenols in tea. Z Lebensm Untersuch Forsch 199:1316


(1994).
Kiehne A and Engelhardt UH, ThermosprayLCMS analysis
of various groups of polyphenols in tea. I. Catechins, avonolO-glycosides and avone-C-glycosides. Z Lebensm Untersuch
Forsch 202:4854 (1996).
Feng Y, McDonald CE and Vick BA, C-Glycosylavones from
hard red spring wheat bran. Cereal Chem 65:452456 (1988).
Feng Y and McDonald CE, Comparison of avonoids in bran
of four classes of wheat. Cereal Chem 66:516518 (1989).
Ramarathnam N, Osawa T, Namiki M and Kawakishi S,
Chemical studies on novel rice hull antioxidants. II.
Identication of isovitexin, a C-glycosyl avonoid. J Agric
Food Chem 37:316319 (1989).
Fossen T, Pedersen AT and Andersen M, Flavonoids from
red onion (Allium cepa). Phytochemistry 47:281285 (1998).
Price KR and Rhodes MJC, Analysis of the major avonol
glycosides present in four varieties of onion (Allium cepa) and
changes in composition resulting from autolysis. J Sci Food
Agric 74:331339 (1997).
Price KR, Colquhoun IJ, Barnes KA and Rhodes MC,
Composition and content of avonol glycosides in green
beans and their fate during processing. J Agric Food Chem
46:48984903 (1998).
Price KR, Casuscelli F, Colquhoun IJ and Rhodes MJC,
Composition and content of avonol glycosides in broccoli
orets (Brassica olearacea) and their fate during cooking. J Sci
Food Agric 77:468472 (1998).
Price KR, Rhodes MJC and Barnes KA, Flavonol glycoside
content and composition of tea infusions made from
commercially available teas and tea products. J Agric Food
Chem 46:25172522 (1998).
Graham HN, Green tea composition, consumption, and
polyphenol chemistry. Prevent Med 21:334350 (1992).
Khokhar S, Venema DP, Hollman PCH, Dekker M and Jongen
WMF, A RP-HPLC method for the determination of tea
catechins. Cancer Lett 114:171172 (1997).
Goldberg DM, Karumanchiri A, Tsang E and Soleas GJ,
Catechin and epicatechin concentrations of red wines:
regional and cultivar-related differences. Am J Enol Vitic
49:2334 (1998).
Risch B and Herrmann K, Die Gehalte an HydroxyzimtsaureVerbindungen und Catechinen in Kern- und Steinobst
[Contents of hydroxycinnamic acid derivatives and catechins
in pome and stone fruit]. Z Lebensm Untersuch Forsch
186:225230 (1988).
Macheix JJ, Fleuriet A and Billot J, Fruit Phenolics. CRC Press,
Boca Raton, FL (1990).
ber das Vorkommen von
Hanefeld M and Herrmann K, U
Proanthocyanidinen, Leukoanthocyanidinen und Catechinen
im Gemuse [On the occurrence of proanthocyanidins,
leucoanthocyanidins and catechins in vegetables]. Z Lebensm
Untersuch Forsch 161:243248 (1976).
Helsper JPFG, Kolodziej H, Hoogendijk JM and Vannorel A,
Characterization and trypsin inhibitor activity of proanthocyanidins from vicia-faba. Phytochemistry 34:12551260
(1993).
Sarkar SK and Howarth RE, Specicity of the vanillin test for
avanols. J Agric Food Chem 24:317320 (1976).
Solich P, Opletal L and Sovova M, Comparison of different
methods for the spectrophotometric determination of
()-epicatechin. Pharmazie 51:954956 (1996).
Khokhar S, Venema D, Hollman PCH, Dekker M and Jongen
W, A RP-HPLC method for the determination of tea
catechins. Cancer Lett 114:171172 (1997).
Bronner WE and Beecher GR, Method for determining the
content of catechins in tea infusions by high-performance
liquid chromatography. J Chromatogr A 805:137142 (1998).
Etievant P, Schlich P, Bertrand A, Symonds P and Bouvier JC,
Varietal and geographic classication of French red wines in

J Sci Food Agric 80:10811093 (2000)

40
41

42

43

44

45

46

47

48

49

50

51

52

53

54
55

56

57

terms of pigments and avonoid compounds. J Sci Food Agric


42:3954 (1988).
Archier P, Coen S and Roggero JP, Phenolic contents of single
variety wines. Sci Alim 12:453466 (1992).
Soleas GJ, Dam J, Carey M and Goldberg DM, Toward the
ngerprinting of wines: cultivar-related patterns of polyphenolic constituents in Ontario wines. J Agric Food Chem
45:38713880 (1997).
Goldberg DM, Tsang E, Karumanchiri A, Diamandis EP,
Soleas G and Ng E, Method to assay the concentrations of
phenolic constituents of biological interest in wines. Anal
Chem 68:16881694 (1996).
Oszmianski J and Sapis JC, Fractionation and identication of
some low molecular weight grape seed phenolics. J Agric Food
Chem 37:12931297 (1989).
Escribano-Bailon MT, Guerra MT, Rivas-Gonzalo JC and
Santos-Buelga C, Proanthocyanidins in skins from different
grape varieties. Z Lebensm Untersuch Forsch 200:221224
(1995).
Santos-Buelga C, Francia-Aricha EM and Escribano-Bailon
MT, Comparative avan-3-ol composition of seeds from
different grape varieties. Food Chem 53:197201 (1995).
Oszmianski J and Lee CY, Isolation and HPLC determination
of phenolic compounds in red grapes. Am J Enol Vitic 41:204
206 (1990).
Van Gorsel H, Li C, Kerbel EL, Smits M and Kader AA,
Compositional characterization of prune juice. J Agric Food
Chem 40:784789 (1992).
Fernandez de Simon B, Perez-Ilzarbe J, Hernandez T, GomezCordoves C and Estrella I, Importance of phenolic compounds for the characterization of fruit juices. J Agric Food
Chem 40:15311535 (1992).
Amiot MJ, Tacchini M, Aubert SY and Oleszek W, Inuence of
cultivar, maturity stage, and storage conditions on phenolic
composition and enzymatic browning of pear fruits. J Agric
Food Chem 43:11321137 (1995).
Donovan JL, Meyer AS and Waterhouse AL, Phenolic
composition and antioxidant activity of prunes and prune
juice (Prunus domestica). J Agric Food Chem 46:12471252
(1998).
Stohr H and Herrmann K, Die phenolischen Inhaltsstoffe des
Obstes. V. Die phenolischen Inhaltsstoffe der Erdbeeren und
deren Veranderungen wahrend Wachstum und Reife der
Fruchte [The phenolics of fruits. V. The phenolics of
strawberries and their changes during development and
ripeness of the fruits]. Z Lebensm Untersuch Forsch 158:341
348 (1975).
Stohr H and Herrmann K, Die phenolischen Inhaltsstoffe des
Obstes. VI. Die phenolischen Inhaltsstoffe der Johannisbeeren, Stachelbeeren und Kulturheidelbeeren. Veranderungen
der Phenolsauren und Catechine wahrend Wachstum und
Reife von Schwarzen Johannisbeeren [The phenolics of fruits.
VI. The phenolics of currants, gooseberries and blueberries.
Changes in phenolic acids and catechins during development
of black currants]. Z Lebensm Untersuch Forsch 159:3137
(1975).
Rommel A and Wrolstad RE, Inuence of acid and base
hydrolysis on the phenolic composition of red raspberry juice.
J Agric Food Chem 41:12371241 (1993).
Ramrez-Martnez JR and Luh BS, Phenolic compounds in
frozen avocados. J Sci Food Agric 24:219225 (1973).
Deshpande SS and Cheryan M, Determination of phenolic
compounds of dry beans using vanillin, redox and precipitation assays. J Food Sci 52:332334 (1987).
Sanbongi C, Osakabe N, Natsume M, Takizawa T, Gomi S and
Osawa T, Antioxidative polyphenols isolated from Theobroma
cacao. J Agric Food Chem 46:454457 (1998).
Hammerstone JF, Lazarus SA, Mitchell AE, Rucker R and
Schmitz HH, Identication of procyanidins in cocoa (Theobroma cacao) and chocolate using high-performance liquid

1091

PCH Hollman, ICW Arts

58
59
60
61

62
63

64

65
66

67
68

69

70
71

72

73
74

75

76

chromatography mass spectrometry. J Agric Food Chem


47:490496 (1999).
Burda S, Oleszek W and Lee CY, Phenolic compounds and
their changes in apples during maturation and cold storage. J
Agric Food Chem 38:945948 (1990).
Mosel HD and Herrmann K, Changes in catechins and
hydroxycinnamic acid derivatives during development of
apples and pears. J Sci Food Agric 25:251256 (1974).
Kim H and Keeney PG, ()-Epicatechin content in fermented
and unfermented cocoa beans. J Food Sci 49:10901092
(1984).
Suarez Valles B, Santamara Victorero J, Mangas Alonso JJ and
Blanco Gomis D, High-performance liquid chromatography
of the neutral phenolic compounds of low molecular weight in
apple juice. J Agric Food Chem 42:27322736 (1994).
Spanos GA, Wrolstad RE and Heatherbell DA, Inuence of
processing and storage on the phenolic composition of apple
juice. J Agric Food Chem 38:15721579 (1990).
Kermasha S, Goetghebeur M, Dumont J and Couture R,
Analyses of phenolic and furfural compounds in concentrated
and non-concentrated apple juices. Food Res Int 28:245252
(1995).
Bengoechea ML, Sancho AI, Bartolome B, Estrella I, Gomezcordoves C and Hernandez MT, Phenolic composition of
industrially manufactured purees and concentrates from
peach and apple fruits. J Agric Food Chem 45:40714075
(1997).
Lee CY and Jaworski A, Phenolic compounds in white grapes
grown in New York. Am J Enol Vitic 38:277281 (1987).
Spanos GA and Wrolstad RE, Inuence of processing and
storage on the phenolic composition of Thompson Seedless
grape juice. J Agric Food Chem 38:15651571 (1990).
Kaneda H, Kano Y, Osawa T, Kawakishi S and Kamimura M,
Effect of free radicals on haze formation in beer. J Agric Food
Chem 38:19091912 (1990).
Achilli G, Cellerino GP, Gamache PH and Melzi d'Eril GV,
Identication and determination of phenolic constituents in
natural beverages and plant extracts by means of a coulometric electrode array system. J Chromatogr 632:111117
(1993).
Madigan D, Mcmurrough I and Smyth MR, Determination of
proanthocyanidins and catechins in beer and barley by highperformance liquid chromatography with dual-electrode
electrochemical detection. Analyst 119:863868 (1994).
Mcmurrough I, Madigan D and Smyth MR, Adsorption by
polyvinylpolypyrrolidone of catechins and proanthocyanidins
from beer. J Agric Food Chem 43:26872691 (1995).
Seto R, Nakamura H, Nanjo F and Hara Y, Preparation of
epimers of tea catechins by heat treatment. Biosci Biotechnol
Biochem 61:14341439 (1997).
Hertog MGL, Hollman PCH, Katan MB and Kromhout D,
Intake of potentially anticarcinogenic avonoids and their
determinants in adults in the Netherlands. Nutr Cancer
20:2129 (1993).
Kuhnau J, The avonoids. A class of semi-essential food
components: their role in human nutrition. World Rev Nutr
Diet 24:117191 (1976).
Hertog MGL, Kromhout D, Aravanis C, Blackburn H, Buzina
R, Fidanza F, Giampaoli S, Jansen A, Menotti A, Nedeljkovic
S, Pekkarinen M, Simic BS, Toshima H, Feskens EJM,
Hollman PCH and Katan MB, Flavonoid intake and longterm risk of coronary heart disease and cancer in the Seven
Countries Study. Arch Intern Med 155:381386 (1995).
Rimm EB, Katan MB, Ascherio A, Stampfer MJ and Willett
WC, Relation between intake of avonoids and risk for
coronary heart disease in male health professionals. Ann
Intern Med 125:384389 (1996).
Hertog MGL, Feskens EJM, Hollman PCH, Katan MB and
Kromhout D, Dietary antioxidant avonoids and risk of
coronary heart disease: the Zutphen Elderly Study. Lancet
342:10071011 (1993).

1092

77 Knekt P, Jarvinen R, Reunanen A and Maatela J, Flavonoid


intake and coronary mortality in Finland: a cohort study. Br
Med J 312:478481 (1996).
78 Hertog MGL, Sweetnam PM, Fehily AM, Elwood PC and
Kromhout D, Antioxidant avonols and ischemic heart
disease in a Welsh population of men: the Caerphilly Study.
Am J Clin Nutr 65:14891494 (1997).
79 Hollman PCH and Katan MB, Absorption, metabolism, and
bioavailability of avonoids, in Flavonoids in Health and
Disease, Ed by Rice-Evans C and Packer L Marcel Dekker,
New York, pp 483522 (1997).
80 Hollman PCH, Tijburg LBM and Yang CS, Bioavailability of
avonoids from tea. Crit Rev Food Sci Nutr 37:719738
(1997).
81 Grifths LA and Barrow A, Metabolism of avonoid compounds in germ-free rats. Biochem J 130:11611162 (1972).
82 Grifths LA and Smith GE, Metabolism of myricetin and
related compounds in the rat. Metabolite formation in vivo
and by the intestinal microora in vitro. Biochem J 130:141
151 (1972).
83 Scheline RR, The metabolism of drugs and other organic
compounds by the intestinal microora. Acta Pharmacol
Toxicol 26:332342 (1968).
84 Grifths LA and Smith GE, Metabolism of apigenin and related
compounds in the rat. Metabolite formation in vivo by the
intestinal microora in vitro. Biochem J 128:901911 (1972).
85 Hollman PCH, de Vries JHM, van Leeuwen SD, Mengelers
MJB and Katan MB, Absorption of dietary quercetin
glycosides and quercetin in healthy ileostomy volunteers.
Am J Clin Nutr 62:12761282 (1995).
86 Nieder M, Pharmakokinetik der Ginkgo-Flavonole im Plasma.
Munch Med Wochenschr 133 (Suppl 1):S61S62 (1991).
87 Hollman PCH, van der Gaag MS, Mengelers MJB, van Trijp
JMP, de Vries JHM and Katan MB, Absorption and
disposition kinetics of the dietary antioxidant quercetin in
man. Free Radic Biol Med 21:703707 (1996).
88 Hollman PCH, van Trijp JMP, Buysman MNCP, van der
Gaag MS, Mengelers MJB, de Vries JHM and Katan MB,
Relative bioavailability of the antioxidant quercetin from
various foods in man. FEBS Lett 418:152156 (1997).
89 Hollman PCH, Buysman MNCP, van Gameren Y, Cnossen
PJ, de Vries JHM and Katan MB, The sugar moiety is a
major determinant of the absorption of dietary avonoid
glycosides in man. Free Radic Res 31:569573 (1999).
90 Day AJ, DuPont MS, Ridley S, Rhodes M, Rhodes MJC,
Morgan MRA and Williamson G, Deglycosylation of
avonoid and isoavonoid glycosides by human small
intestine and liver b-glucosidase activity. FEBS Lett 436:71
75 (1998).
91 Aziz AA, Edwards CA, Lean MEJ and Crozier A, Absorption
and excretion of conjugated avonols, including quercetin-4'O-b-glucoside and isorhamnetin-4'-O-b-glucoside by human
volunteers after the consumption of onions. Free Radic Res
29:257269 (1998).
92 Mizuma T, Ohta K and Awazu S, The b-anomeric and glucose
preferences of glucose transport carrier for intestinal active
absorption of monosaccharide conjugates. Biochim Biophys
Acta 1200:117122 (1994).
93 Gee JM, DuPont MS, Rhodes MJC and Johnson IT, Quercetin
glucosides interact with the intestinal glucose transport
pathway. Free Radic Biol Med 25:1925 (1998).
94 Walgren RA, Walle UK and Walle T, Transport of quercetin
and its glucosides across human intestinal epithelial Caco-2
cells. Biochem Pharmacol 55:17211727 (1998).
95 Conquer JA, Maiani G, Azzini E, Raguzzini A and Holub BJ,
Supplementation with quercetin markedly increases plasma
quercetin concentration without effect on selected risk factors
for heart disease in healthy subjects. J Nutr 128:593597
(1998).
96 Manach C, Morand C, Crespy V, Demigne C, Texier O,
Regerat F and Remesy C, Quercetin is recovered in human

J Sci Food Agric 80:10811093 (2000)

Flavonols, avones and avanols


plasma as conjugated derivatives which retain antioxidant
properties. FEBS Lett 426:331336 (1998).
97 Serani M, Ghiselli A and Ferro-Luzzi A, In vivo antioxidant
effect of green and black tea in man. Eur J Clin Nutr 50:2832
(1996).
98 Shimoi K, Okada H, Furugori M, Goda T, Takase S, Suzuki M,
Hara Y, Yamamoto H and Kinae N, Intestinal absorption of
luteolin and luteolin 7-O-beta-glucoside in rats and humans.
FEBS Lett 438:220224 (1998).
99 Balant L, Burki B, Wermeille M and Golden G, Comparison of
some pharmacokinetic parameters of ()-cyanidanol-3 obtained with specic and non-specic analytical methods.
Arzneim Forsch 29:17581762 (1979).
100 Giles AR and Gumma A, Biopharmaceutical evaluation of
cyanidanol tablets using pharmacokinetic techniques. Arzneim Forsch 23:98100 (1973).
101 Yang CS, Chen L, Lee M-J, Balentine D, Kuo MC and Schantz
SP, Blood and urine levels of tea catechins after ingestion of
different amounts of green tea by human volunteers. Cancer
Epidemiol Biomark Prevent 7:351354 (1998).
102 Van het Hof KH, Kivits GAA, Weststrate JA and Tijburg
LBM, Bioavailability of catechins from tea: the effect of milk.
Eur J Clin Nutr 52:356359 (1998).
103 Kivits GAA, van der Sman FJP and Tijburg LBM, Analysis of
catechins from green and black tea in humans: a specic and
sensitive colorimetric assay of total catechins in biological
uids. Int J Food Sci Nutr 48:387392 (1997).
104 Maiani G, Serani M, Salucci M, Azzini E and Ferro-Luzzi A,
Application of a new high performance liquid chromatographic method for measuring selected polyphenols in human
plasma. J Chromatogr B 692:311317 (1997).
105 Nakagawa K and Miyazawa T, Chemiluminescence high
performance liquid chromatographic determination of tea
catechin, ()-epigallocatechin-3-gallate, at picomole levels in
rat and human plasma. Anal Biochem 248:4149 (1997).

J Sci Food Agric 80:10811093 (2000)

106 Lee M-J, Wang Z-Y, Li H, Chen L, Sun Y, Gobbo S, Balentine


DA and Yang CS, Analysis of plasma and urinary tea
polyphenols in human subjects. Cancer Epidemiol Biomark
Prevent 4:393399 (1995).
107 Richelle M, Tavazzi I, Enslen M and Offord EA, Plasma
kinetics in man of epicatechin from black chocolate. Eur J
Clin Nutr 53:2226 (1999).
108 Van het Hof KH, Wiseman SA, Yang CS and Tijburg LBM,
Plasma and lipoprotein levels of tea catechins following
repeated tea consumption. Proc Soc Exp Biol Med 220:203
209 (1999).
109 Scheline RR, Metabolism of foreign compounds by gastrointestinal microorganisms. Pharmacol Rev 25:451523
(1973).
110 Hattori M, Shu YZ, El-Sedawy Al, Namba T, Kobashi K and
Tomimori T, Metabolism of homoorientin by human
intestinal bacteria. J Nat Prod 51:874878 (1988).
111 Rice-Evans CA, Miller NJ and Paganga G, Structureantioxidant activity relationships of avonoids and phenolic acids.
Free Radic Biol Med 20:933956 (1996).
112 Nakagawa K, Okuda S and Miyazawa T, Dose-dependent
incorporation of tea catechins, ()-epigallocatechin-3-gallate
and ()-epigallocatechin, into human plasma. Biosci Biotechnol Biochem 61:19811985 (1997).
113 Ricardo da Silva JM, Rosec JP, Bourzeix M and Heredia N,
Separation and quantitative determination of grape and wine
procyanidins by high performance reversed phase liquid
chromatography. J Sci Food Agric 53:8592 (1990).
114 Andrade P, Seabra R, Ferreira M, Ferreres F and Garciaviguera
C, Analysis of non-coloured phenolics in port wines by
capillary zone electrophoresisinuence of grape variety and
ageing. Z Lebensm Untersuch Forsch 206:161164 (1998).
115 Gugler R, Leschik M and Dengler HJ, Disposition of quercetin
in man after single oral and intravenous doses. Eur J Clin
Pharmacol 9:229234 (1975).

1093