Proceedings of the Third International Scientific Symposium

on Tea and Human Health: Role of Flavonoids in the Diet

Metabolism of Tea Flavonoids in the Gastrointestinal Tract1,2

Jeremy P. E. Spencer3
Wolfson Centre for Age-Related Disease, GKT School of Biomedical Science, King’s College London,
London, SE1 9RT, UK

ABSTRACT There is considerable interest in the bioavailability of flavan-3-ols such as tea catechins and their
bioactivity in vivo. Although flavanols such as catechin and epicatechin have long been characterized as powerful
antioxidants in vitro, evidence suggests that these compounds undergo significant metabolism and conjugation
during absorption in the small intestine and in the colon. In the small intestine these modifications lead primarily to
the formation of glucuronide conjugates that are more polar than the parent flavanol and are marked for renal
excretion. Other phase II processes lead to the production of O-methylated forms that have reduced antioxidant
potential via the methylation of the B-ring catechol. Significant modification of flavanols also occurs in the colon
where the resident microflora degrade them to smaller phenolic acids, some of which may be absorbed. Cell,

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animal and human studies have confirmed such metabolism by the detection of flavanol metabolites in the
circulation and tissues. This review will highlight the major sites of flavanol metabolism in the gastrointestinal tract
and the processes that give rise to potential bioactive forms of flavan-3-ols in vivo. J. Nutr. 133: 3255S-3261S,
2003.

KEY WORDS: ● flavonoid ● metabolism ● catechin ● tea ● GI tract

Flavonoids have been the center of huge research interest
over the last decade (1,2). They are the most abundant poly-
phenols in the human diet and are divided into six main
classes based on the degree of oxidation of the C-ring, the
hydroxylation pattern of the ring-structure and the substitu-
tion in the 3-position: flavanols (e.g., epicatechin), flavonols
(e.g., quercetin), flavones (e.g., luteolin), flavanones (e.g.,
naringenin), isoflavones (e.g., genistein) and anthocyanidins
(e.g., cyanidin) (3). In both black and green tea the major
class of flavonoids are the flavanols, which include catechin,
epicatechin, epigallocatechin (EGC),4 epicatechin gallate
(ECG) and epigallocatechin gallate (EGCG). A large number
of in vitro studies has characterized flavanols as powerful
antioxidants capable of efficient scavenging of both reactive
oxygen and reactive nitrogen species (3– 8). The mechanism
1
Presented as part of “The Third International Scientific Symposium on Tea
and Human Health: Role of Flavonoids in the Diet,” given at the United States
Department of Agriculture, September 23, 2002. This conference was sponsored
by the American Cancer Society, American College of Nutrition, American Health
Foundation, American Society for Nutritional Sciences, Food and Agriculture
Organization, and the Linus Pauling Institute at Oregon State University and was
supported by a grant from the Tea Council of the U.S.A. Guest editor for this
symposium was Jeffrey Blumberg, PhD, Jean Mayer USDA Human Nutrition
Research Center on Aging, Tufts University, Boston, MA 02111.
2
This research was supported by the Biotechnology and Biological Sciences
Research Council, Swindon, UK (Grant no. 18/D14751).
3
To whom correspondence should be addressed.
E-mail: jeremy.spencer@kcl.ac.uk.
4
Abbreviations used: COMT, catechol-O-methyl-transferase; ECG, epicat-
echin gallate; EGC, epigallocatechin; EGCG, epigallocatechin gallate; GI, gastro-
intestinal; MRP-2, multidrug resistance-associated protein 2; UGT, UDP-glucu-
rosyltransferase.
of their action as radical scavengers involves the donation of
a hydrogen atom and/or electron to stabilize the radical species
(3). Until recently, the ability of flavanols, and indeed other
flavonoids, to act as classical H-donating antioxidants was
believed to underlie many of their reported health effects
(9 –15). However, it is now clear that their ultimate antioxi-
dant potential, and indeed their resulting potential bioactivity,
in vivo is dependent on the absorption, metabolism, distribu-
tion and excretion of these compounds within the body after
ingestion and the reducing properties of the resulting metab-
olites. An understanding of the processes involved in the
absorption and distribution of tea flavonoids is essential for
determining their potential bioactivities in vivo and their
overall significance in disease prevention. Much data now
exists to support the biotransformation of flavanols and other
flavonoids in the small intestine and colon of the gastrointes-
tinal (GI) tract (10,16 –22), as well as hepatic metabolism
(23–26). This review will attempt to highlight the main sites
of biotransformation of tea flavanols within the GI tract, the
major metabolites generated in the small and large intestine
and the implications of this modification in determining how
flavonoids may act in vivo.
Modification of flavanols in the upper GI tract
Effects of saliva. Few studies exist on the ability of saliva
and gastric juice to alter the flavonoid structure. Saliva has
been found to have little effect on the stability of green tea
catechins (27). For example, when mouth rinsing was per-
formed with an aqueous solution of green tea extract (5.0 g/L)

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3255S
3256S SUPPLEMENT
containing eight catechins, it was observed that each catechin
was retained at ␮g/ml levels in saliva for up to 60 min.
However, degalloylation of flavanol gallate esters in human
saliva has been observed, such as the conversion of EGCG to
EGC in the oral cavity with both subsequently being absorbed
through the oral mucosa (28). A catechin esterase activity
that converts EGCG to EGC has been found in the saliva and
is likely of human origin. However, a common human esterase
inhibitor did not inhibit the activity. Similar incubation of
procyanidin oligomers (dimer-hexamer) in human saliva for
up to 30 min does not result in modification of the compounds
or their decomposition into smaller oligomeric units (22)
suggesting that these compounds remain intact in the mouth
and esophagus before entering the stomach.
Another consideration in the upper GI tract is the possible
interaction of flavanols and procyanidins with salivary pro-
teins. (⫹)-Catechin has been observed to have a higher affin-
ity for salivary proline-rich proteins than (⫺)-epicatechin,
which highlights the importance of the stereochemistry of
flavan-3-ols on their interaction with proteins (29). Further-
more, procyanidin dimers linked through a C(4)-C(8) inter-
flavanoid bond had a greater affinity for proline-rich proteins
than those linked C(4)-C(6). In addition, esterification of a
galloyl group to the C(3) hydroxyl function of (⫺)-epicat-
echin or to the epicatechin moiety of procyanidin dimer B2
had the effect of increasing the binding affinity to proline-rich
proteins (29). It is believed that these specific polyphenol-
protein interactions in the form of adsorption with high-
molecular-weight salivary proteins, bacterial cells and mucous
materials may be one explanation for the observed decrease in
quercetin mutagenicity after incubation with saliva (30).
Modification in the gastric lumen. Although it seems that
monomeric flavan-3-ols are stable in the gastric lumen, flava-
nol oligomers ranging from a dimer to decamer, such as those
isolated from Theobroma cacao, have been observed to be
unstable under conditions of low pH similar to that present in
the gastric juice of the stomach (31). Upon incubation of the
procyanidins with simulated gastric juice (0.1–3 h), oligomers
rapidly decompose essentially to epicatechin monomeric and
dimeric units but also to other oligomeric units at later time
points (31). In contrast to this, a recent investigation in
humans has suggested that there is no cleavage of procyanidins
in the stomach after ingestion of a cocoa beverage (32).
However, chemically the decomposition of procyanidins in
mild acidic environments is well characterized (33) and con-
sequently, if conditions of pH in the stomach are suitably low,
there will be a rapid cleavage of procyanidins to yield smaller
flavanol units that will enter the small intestine.
Metabolism and conjugation in the small intestine
There are many factors that influence the extent and rate of
absorption of ingested compounds by the small intestine (34).
These include physiochemical factors such as molecular size,
lipophilicity, solubility, pKa and biological factors including
gastric and intestinal transit time, lumen pH, membrane per-
meability and first pass metabolism (35,36). Generally fla-
vonoids are present in plants conjugated to sugars and it is
these glycosides that are ingested in the diet and enter the GI
tract. Flavan-3-ols are the exception to this rule and are almost
always present in the diet in the nonglycosylated form (3).
Consequently, unlike other dietary flavonoids, there is no
initial requirement for ␤-glucosidase action prior to absorp-
tion.
Effects of intestinal juice. Upon transfer from the stomach
to the jejunum (the top two-fifths of the small intestine) the
pH rises from about 2 to 7. It is well known that polyphenolic
compounds such as those with catechol structures oxidize in
neutral and alkaline pH environments. EGCG has been ob-
served to rapidly oxidize in authentic intestinal fluid (mea-
sured pH of 8.5) with the amount of EGCG decreasing 81.6%
in only 5 min (37), whereas a similar incubation in murine
plasma (pH 7.4) resulted in only a 29.3% decrease in amount.
However, the oxidation of EGCG resulted in the formation of
dimerized products, which were observed to possess greater
superoxide radical scavenging activity than EGCG itself and
had powerful iron chelating properties (37). Another factor to
consider may be the relative abilities of these polyphenols to
bind to proteins in the food matrices in question. In complex
food matrices, the pH is likely to be buffered for long periods
of time and therefore, oxidation of the flavanols may only
occur to a limited extent during intestinal transit. In addition,
it has been suggested that ascorbate significantly increases the
stability of flavanols incubated in intestinal fluid (38) and
therefore the presence of ascorbate in vivo may stabilize the
polyphenols in the neutral or alkaline environment of the
small intestine.
Jejunal and ileal transfer and metabolism. Many studies
have indicated that significant transfer of ingested flavonoids
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occurs from the lumen of the small intestine to the mesenteric
circulation and that extensive metabolism and conjugation of
the flavonoid occurs during this transfer (2,39 – 46). Although
both Caco-2 cell models and isolated preparations of rat small
intestine (described below and in Fig. 1) (47) have been
utilized to study absorption and metabolism of tea flavanols in
the small intestine, only the latter has provided in-depth
information on events occurring in both the jejunum and
ileum (40,42,46,48 –50). This model allows study of the intes-
tinal transfer of dietary polyphenols (and their glycosides) and
can be used to assess the rate of absorption from the lumen.
The solute under study appears on the serosal surface in the
same form as if it were transferred to the mesenteric circulation
and therefore enterocyte metabolism of flavanols and procya-
nidins, as well as their rate of transfer across specific gut
regions, can be studied (42). Tissue viability is assessed by
measurement of glucose transfer (47) and viability is confirmed
by the finding that fluid transfer continues at a constant rate
for the 90-min collection period and that glucose concentra-
FIGURE 1 Diagramatic representation of the isolated rat small
intestine perfusion model. Buffers and compound are perfused for up to
90 min at a rate of 8 mL/min at 37°C using a peristaltic pump. Serosal
fluid is collected from the bottom of the paraffin chamber and analyzed
by HPLC and mass spectrometry.
 METABOLISM OF TEA FLAVONOIDS IN THE GI TRACT
tion in the absorbed fluid is over double that initially present
in the perfused buffer (42).
Isolated small intestine model. Absorption studies utilizing
this model have been performed with a wide range of fla-
vonoids, their glycosides and hydroxycinnamates. These show
that there is in almost all cases extensive enterocytic metab-
olism of the polyphenol in the enterocyte during transfer from
the luminal to the serosal side (42). The extent of glucu-
ronidation in these experiments was dependent on the fla-
vonoid structure, in that the flavonoids with a substituted
hydroxyl group on the B-ring (i.e., hesperetin) were less pre-
disposed to glucuronidation, whereas the flavonoids contain-
ing a 3⬘,4⬘-ortho-dihydroxy (or catechol) B-ring were trans-
ferred predominantly as glucuronides (42,51). In addition,
monophenolic B-ring flavonoids were also extensively glucu-
ronidated, in particular naringenin, which was only detected
in serosal fluid as naringenin-7-glucuronide (42). Similar pat-
terns of metabolism have been observed in the ileum, although
in general, glucuronidation occurs to a lesser extent most
probably due to lower levels of phase I and II enzymes present
in the ileum compared with the jejunum. Interestingly, two
UDP-glucuronosyltransferase (UGT) isoforms, UGT1A8 and
UGT1A10, of human intestinal mucosa, which are absent in
liver, have been identified by RT-PCR (39).
Although the major metabolites observed on the serosal
side after perfusion of the jejunum with catechin or epicat-
echin were always glucuronidated (at the 5- and 7- positions
on the A-ring), there were also high levels of both O-meth-
ylated and O-methylated-glucuronide forms (Fig. 2) (22,49).
3⬘-O- and 4⬘-O-methylated derivatives of the flavanols were
detected at high levels in the serosal fluid (⬃30% of total
transferred) and together with O-methyl-glucuronidated forms
were the predominant metabolites detected in the serosal fluid
(⬃50%) suggesting that these are the most bioavailable forms
in the small intestine (Fig. 2). As with the other flavonoids
tested in this model, there was a lower level of metabolism
occurring in the ileum although the total amounts of unme-
tabolised catechin and epicatechin absorbed were significantly
FIGURE 2 The structures of the main small intestinal metabolites
of epicatechin produced in the isolated rat small intestine perfusion
model. (A) 3⬘-O-methyl epicatechin; (B) 4⬘-O-methyl epicatechin; (C)
3⬘-O-methyl epicatechin-5-glucuronide; (D) epicatechin-5-glucuronide;
(E) epicatechin-7-glucuronide.
3257S
higher than in the jejunum (42). The greater susceptibility of
flavanols over other flavonoids to methylation in the jejunum
may reside in the specificity of catechol-O-methyl-transferase
(COMT) for these compounds (52). These data have been
supported in a similar model, in which the rat jejunum and
ileum were perfused with catechin (41). In this study, catechin
was absorbed into intestinal cells and metabolized extensively
to a point where no native catechin could be detected in
plasma from the mesenteric vein. Mesenteric plasma con-
tained glucuronide conjugates of catechin and 3⬘-O-methyl
catechin, indicating the intestinal origin of these conjugates
and the large role the small intestine plays in the biotransfor-
mation of flavanols during absorption (41,49). Although many
studies have identified flavanol metabolites as being the main
forms found entering the hepatic portal vein after absorption
from the small intestine, the native flavanols are detected in
small amounts (42,49). For example, oral administration of the
tea catechins, epicatechin, EGC, ECG and EGCG, to rats led
to the detection of all four flavanols in the portal blood (53)
clearly indicating that these flavonoids may be absorbed intact
to a small degree.
Studies have also shown that catechin and tannic acid may
interact with the small intestine but only catechin appears
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able to traverse the gut (46). This may be due to the binding
of tannic acid and catechin by endogenous proteins in the
intestinal lumen limiting their absorption in the small intes-
tine. The binding of flavanols to proteins in the small intestine
provides one explanation for their relatively low absorption.
However, it should also be noted that the protein-binding
properties of catechins may be linked to their bactericidal
abilities and may play several roles in the digestive tract. In the
small intestine, catechins inhibit alpha-amylase activity but do
not affect lactic acid bacteria (54). The inclusion of tea
catechins in the diet for several weeks had the effect of
decreasing putrefactive products and at the same time in-
creased organic acids by lowering pH (54). The high-molec-
ular-weight procyanidins have a high affinity for proteins and
their absorption through the gut barrier is most likely limited
to lower oligomeric forms and to the metabolites formed by the
colonic microflora. Recently, perfusion of isolated small intes-
tine with the procyanidin dimers B2 and B5 extracted from
cocoa indicated that both forms of dimer are transferred to the
serosal side of enterocytes but only to a very small extent
(⬍1% of the total transferred flavanol-like compounds) (55).
Perfusion of dimer mainly resulted in large amounts of unme-
tabolised/unconjugated epicatechin monomer being detected
on the serosal side (⬃95.8%). Low levels of O-methylated
dimer were also detected (⬃3.2%), but no conjugates and
metabolites of epicatechin indicating that metabolism of
monomer and dimer is limited during dimer cleavage/translo-
cation.
Caco-2 cell model. Another approach to obtaining a better
understanding of the bioavailability of flavonoids and their
absorption and metabolism in the small intestine has been in
the use of cultured human Caco-2 cells. In recent years studies
with the human Caco-2 cell line have helped to unravel the
complex processes involved in the absorption and metabolism
of dietary polyphenols in humans. These cells were originally
derived from human colonic adenocarcinoma cells. However,
despite their origin, after culture in DMEM supplemented with
fetal calf serum (10%) and glutamate (20 mmol/L) for
⬃14 –21 d (37°C; 5% CO2) these cells more closely resemble
enterocytes than colonocytes both morphologically and bio-
chemically. Fully differentiated Caco-2 monolayers also resem-
ble enterocytes in that they express transport systems for
sugars, amino acids, bile acids and dipeptides and express many
3258S SUPPLEMENT
brush border membrane enzymes, including aminopeptidase,
alkaline phosphatase, sucrase and ␥-glutamyltranspeptidase.
Even so, it is important to consider the full enzymatic profile
of the Caco-2 cell model to be used because they may not be
identical to normal human small intestinal cells in terms of
metabolizing enzymes such as cytochrome P450s.
Absorption studies with flavanols and procyanidins using
Caco-2 cells are few (56 –58). Interestingly, apical to basolat-
eral transfer of epicatechin across the Caco-2 monolayer has
not been detected thus far (56). It is believed that this is due
to the action of multidrug resistance-associated protein-2
(MRP2) which acts to rapidly efflux epicatechin and epicat-
echin metabolites from the cell interior back out to the apical
side. Other experiments with radiolabeled procyanidins have
shown that flavanol dimers and trimers were transferred to the
same extent as epicatechin monomer, whereas oligomers with
an average degree of polymerization of 7 were not (23,59). In
contrast, we observed very little transfer of the dimer across
Caco-2 cells, which was in agreement with our studies in the
isolated rat small intestine model (55).
The model has mainly been utilized for the study of the
absorption and metabolism of other flavonoids in the small
intestine and the data from this work has helped support the
general concept of the extensive metabolism of flavonoids in
the small intestine. For example, Galijatovic et al. (60,61)
observed that chrysin and quercetin are glucuronidated on
transfer and also induce UGT in Caco-2 cells. Exposure of the
cells to chrysin (50 ␮mol/L; 2 h) resulted in a 3.8-fold increase
in chrysin glucuronidation and a 38% decrease in sulfation in
intact cells. Induction of phase II enzymes, such as UGT, may
be important for the bioavailability of carcinogens and other
toxic chemicals by increasing the ease and rate at which they
are excreted from the body. In addition, Wahlgren et al.
(62,63) studied the absorption of the predominant dietary
form of quercetin, quercetin-4⬘-␤-glucoside. Although querce-
tin itself was not transferred, they found that the transport of
the glucoside involved apical MRP2 suggesting that the trans-
fer of glycosides may occur in the small intestine.
Colonic metabolism
Studies have suggested that the extent of absorption of
dietary polyphenols in the small intestine is relatively small
(10 –20%) (42,48,49). The implications of this low absorption
in the small intestine is that the majority of ingested polyphe-
nols, including those absorbed and conjugated in the entero-
cytes and/or the liver before transport back out into the lumen
either directly or via the bile (40), will reach the large intes-
tine where they encounter the colonic microflora. The colon
contains ⬃1012 microorganisms/cm3, which have an enormous
catalytic and hydrolytic potential, and this enzymatic degra-
dation of flavonoids by the colonic microflora results in a huge
array of new metabolites. For example, bacterial enzymes may
catalyze many reactions including hydrolysis, dehydroxylation,
demethylation, ring cleavage and decarboxylation as well as
rapid deconjugation (24). Unlike human enzymes, the micro-
flora catalyze the breakdown of the flavonoid backbone itself
to simpler molecules such as phenolic acids. Specific metabo-
lites have been observed in urine after consumption of a
variety of phenolics. For example, the glycine conjugate of
benzoic acid, hippuric acid, is primarily derived from plant
phenolics and aromatic amino acids through the action of
intestinal bacteria and, consequently, the level of hippuric
acid would be expected to increase in the urine of individuals
consuming diets rich in flavanols or polyphenols in general. It
must be noted, however, that hippuric acid may possibly derive
from other sources such as quinic acid or, in quantitative
terms, more importantly from the aromatic amino acids tryp-
tophan, tyrosine and phenylalanine, as well as from the use of
benzoic acid as a food preservative. One investigation in
humans suggests an association between black tea consump-
tion and excreted amounts of hippuric acid (64). The forma-
tion of hippuric acid and hydroxyhippuric acids seems to be a
possible central metabolic pathway for dietary flavonoids, in
which the colon microflora and the liver are active metabolic
sites (24). Other hydroxybenzoic acid glycine derivatives such
as 4-hydroxyhippuric acid, vanilloylglycine, isovanilloylgly-
cine might also appear in reasonable amounts in urine after
polyphenol consumption in general.
The 5,7,3,3⬘,4⬘-hydroxylation pattern of flavan-3-ols is be-
lieved to enhance ring opening after hydrolysis (22,24) and
metabolism of flavanols by enzymes of the microflora of the
large intestine results in many metabolites: 3,4-dihydrophenyl-
acetic acid, 3-hydroxyphenylacetic acid, homovanillic acid
and their conjugates derived from the B-ring (24) and phe-
nolic acids from the C-ring. Flavanols, because of their struc-
tures (no C-4 carbonyl group), can also degrade to the specific
metabolites phenylvalerolactones. Phenylpropionic acids
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(which may undergo further metabolism to benzoic acids) may
also be the products of flavanol metabolism in animal studies,
which demonstrates fission of the A-ring (24). Only 3.1% of
the ingested catechin in rats was extractable from feces, indi-
cating that major absorption and/or degradation of catechin
had occurred in the GI tract (65). Such metabolites of fla-
vanols have been detected in human plasma and urine after a
single ingestion of green tea (66) which suggests that there
may be significant metabolism by gut microflora in the colon.
The two metabolites, (⫺)-5-(3⬘,4⬘,5⬘-trihydroxyphenyl)-␥-
valerolactone and (⫺)-5-(3⬘,4⬘-dihydroxyphenyl)-␥-valero-
lactone were identified in urine by both LC-MS/MS and
NMR, appearing 7.5–13.5 h after ingestion (after a 3 h lag
time), whereas epicatechin and EGC peaked at 2 h. In addi-
tion to their late excretion profiles, the amounts of metabolite
excreted were 8 –25-fold greater than that of epicatechin and
EGC and accounted for 6 –39% of the epicatechin and EGC
ingested. The late excretion and high levels of these metabo-
lites would suggest that they are generated from the precursors
epicatechin and EGC by the colonic microorganisms. Previous
to this human study, similar observations were made in rats fed
with labeled catechin (67). Here catechin glucuronides were
observed in the bile after dosing rats with the catechin and m-
and p-hydroxyphenylproprionic acid, ␦-(3-hydroxyphenyl)-␥-
valerolactone and ␦-(3,4-dihydroxyphenyl)-␥-valerolactone
were identified as metabolites arising due to the action of the
colonic microflora (67).
The metabolism of procyanidins by human colonic micro-
flora has been studied in vitro under anoxic conditions using
nonlabeled and (14)C-labeled purified proanthocyanidin poly-
mers (68). Interestingly, the oligomers were almost totally
degraded after 48 h of incubation and meta- or para-monohy-
droxylated-phenylacetic, phenylpropionic and phenylvaleric
acids were identified as metabolites, providing the first evi-
dence that dietary flavan-3-ol polymers can be degraded to
low-molecular-weight aromatic compounds in the body (68).
Action of flavanol metabolites in cell systems
The action of flavonoid metabolites, in particular the O-
methylated flavonoids deriving from small intestinal absorp-
tion, are now of great current interest. For example, the ability
 METABOLISM OF TEA FLAVONOIDS IN THE GI TRACT
of 3⬘-O-methyl epicatechin and epicatechin glucuronides to
protect against apoptotic cell death induced by hydrogen per-
oxide or oxidized LDL has been investigated (69 –71). A
mixture of epicatechin-5-O-␤-D- and epicatechin-7-O-␤-D-
glucuronides exhibited no significant protection against per-
oxide-induced loss in neuronal or fibroblast viability and also
failed to prevent peroxide-induced caspase-3 activation in
both cell models (71). This lack of activity may be based on
the increased polarity of the glucuronide and its reduced
ability to partition, which would limit its access to cells. This
is consistent with the observation that uptake of epicatechin
glucuronide into both cortical neurons and dermal fibroblasts
was not detectable (71). Another possibility is that the A-ring
is the structurally important feature for cell recognition or
biological action within cells and in vivo and presence of a
bulky glucuronide on the A-ring limits its activity. In contrast,
epicatechin and 3⬘-O-methyl epicatechin, have been shown to
elicit strong cytoprotective effects in fibroblasts and neurons
associated with both cell types (69,71,72). Interestingly, the
3⬘-O-methyl-epicatechin metabolite was as effective as epicat-
echin at preventing damage and was also equally effective in
protecting neurons against oxLDL-induced activation of c-jun
N-terminal kinase, c-jun, and procaspase-3 (70). The ability of
the O-methylated metabolite to exert protection similar to the
native epicatechin is interesting because the H-donating po-
tential of the two compounds is very different, with the O-
methylated compound having a reduced capacity to donate
electrons from its B-ring. This evidence would suggest that
flavonoids might function to protect cells against death in-
duced by oxidants by a mechanism independent of its classical
antioxidant properties, a concept which is beginning to re-
ceive much attention. However, it is also possible that in vivo
demethylation may occur intracellularly by the action of cy-
tochrome P450 enzymes cleaving the O-methylated metabolite
to epicatechin.
Additional studies have identified the 5-O-␤-D-glucuronide
of catechin and epicatechin excreted in the urine of rats
postingestion. The metabolism to a glucuronide does not in-
terfere with their antioxidant properties as assessed by their
ability to scavenge superoxide (26,73) suggesting that in
plasma they may still act as antioxidants. Although glucu-
ronides do not enter cells readily, it is also possible that they
might be cleaved by the action of ␤-glucuronidases located in
human tissues such as the liver (74) or by neutrophils that
release ␤-glucuronidases when activated (51,75). For example,
various quercetin glucuronides have been shown to be decon-
jugated by liver cell-free extracts and by pure recombinant
human ␤-glucuronidase indicating that the cleavage of glucu-
ronides to free aglycones may occur in vivo (74). In terms of
other possible modes of action, studies suggest that the me-
tabolism of flavonoids by phase I and II enzymes in the small
intestine may aid detoxification of potential carcinogens. For
example, alphanapthoflavone increases the activity of
P450/3A in human jejunal and ileal microsomes (76) and
chrysin causes an induction of intestinal UGT in Caco-2 cells
(60,77).
There is also a need to assess the role of the colonic
microflora in the overall bioavailability and potential bioac-
tivity of dietary flavonoids. The extent of absorption of colonic
metabolites is unclear at this time and there is a growing
interest in the potential effects of the phenolic acids and their
derivatives as potentially beneficial agents. For example, the
human intestinal bacteria metabolites of rutin and quercetin,
3,4-dihydroxyphenylacetic acid and 4-hydroxyl-phenylacetic
acid have been shown to possess a more effective antiplatelet
3259S
aggregation activity than rutin and quercetin (78). Further-
more, 2,4,6-trihydroxybenzaldehyde and quercetin were more
effective than rutin on the cytotoxicity against tumor cell
lines. In addition, the effects the phenolics themselves have on
the microflora is an emerging field and it is possible that a
flavanol-induced change in the rich colonic bacterial popula-
tion may have an influence on the overall health of an
individual, the so called prebiotic concept.
Conclusions
Over the recent years we have gained a greater knowledge
of the bioavailable metabolites of dietary flavonoids and it is
now essential to fully evaluate the role of these conjugates and
metabolites in disease prevention (Fig. 3). It is clear that the
GI tract plays a very significant role in the metabolism and
conjugation of these polyphenols before the liver is reached. In
the jejunum and ileum of the small intestine there is efficient
glucuronidation of flavanols by the action of UGT enzymes
and extensive O-methylation by the action of COMT. Unab-
sorbed flavanols, and those taken up, metabolized in the small
intestine and liver and transported back into the intestinal
lumen, will reach the large intestine where they are further
Downloaded from jn.nutrition.org by on October 11, 2010
metabolized by the gut microflora into smaller phenolic acids
and valerolactones. The extent to which these phenolic acids
are absorbed in the colon is presently unclear. However, they
are detected in plasma and are often further conjugated and
metabolized in the liver. Remaining compounds derived from
flavonoid intake pass out in the feces. It is now important to
assess whether the observed metabolism aids entry into cells
and/or renders them better or worse at providing protection
against different stresses, such as oxidative or nitrative stress.
New data in the field is already beginning to suggest that
flavonoids may act to protect cells by more complex mecha-
nisms than was once thought (70). Eventually it is hoped that
FIGURE 3 Summary of the formation of metabolites and conju-
gates of flavonoids in humans. Cleavage of procyanidins may occur in
the stomach in environments of low pH. All classes of flavonoids
undergo extensive metabolism in the jejunum and ileum of the small
intestine and the resulting metabolites enter the portal vein and un-
dergo further metabolism in the liver. Colonic microflora degrade fla-
vonoids into smaller phenolic acids, which may also be absorbed. The
fate of most of these metabolites is renal excretion, however, the extent
to which these compounds enter cells and tissues is unknown.
3260S SUPPLEMENT
these studies will enable specific dietary recommendations to
be made which will increase general health in the population.
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