bioavailability of anthocyanins and derivatives

Iva Fernandesa, Ana Fariaa,b,c, Conceicao Calhaub,d, Victor de Freitasa, Nuno Mateusa,*

aChemistry Investigation Centre (CIQ), Department of Chemistry and Biochemistry, Faculty of Sciences, University of Porto,

Rua do Campo Alegre, 4169-007 Porto, PortugbDepartment of Biochemistry (U38-FCT), FacucFaculty of Nutrition and Food Sciences, UnivedCINTESIS Center for Research in Health Te

4200-319 Porto, Portugal

A R T I C L E I N F O

Article history:

Available online xxxx

ural pigments in the plant kingdom, thereby constituting a

part of the world natural heritage. They confer a great diversity

of colours, touching practically all visible spectra, from or-

ange and red through purple and blue hues.

ity has be

understa

different

structures from numerous natural sources have bee

terized and their physicalchemical properties determined

(Deroles, 2009); their biosynthesis pathways have been eluci-

dated, and new plants with a la carte colours created by

J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x x

Avai lab le a t www.sc ienced i rec t .com

w.* Corresponding author. Tel.: +351 220402562.1. Introduction

Anthocyanins are one of the most widespread families of nat-

Over the years, the scientific commun

ing on these amazing molecules trying to

cyanins and their properties. Many1756-4646/$ - see front matter 2013 Elsevier Ltd. All rights reserved.http://dx.doi.org/10.1016/j.jff.2013.05.010

E-mail address: [email protected] (N. Mateus).

Please cite this article in press as: Fernandes, I. et al., Bioavailability of anthocyanins and derivatives, Journal of Functional Foods (201dx.doi.org/10.1016/j.j.2013.05.010en focus-

nd antho-

chemical

n charac-sensitivity of the analytical method, the possible ingestion of pigments (anthocyanin deriv-

atives, especially in the case of red wine) and the influence of the food matrix. Generally,

the bioavailability of anthocyanins is presumed but whether the effect is due to the native

compounds or other forms, which mechanism are involved or which factors have crucial

impact on bioavailability still remain underexplored.

2013 Elsevier Ltd. All rights reserved.membranes, the effecKeywords:

Absorption

Anthocyanins

Bioavailability

Metabolism

Microbiotaal

lty of Medicine, University of Porto, Al. Prof. Hernani Monteiro, 4200-319 Porto, Portugal

rsity of Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal

chnologies and Information Systems, University of Porto, Al. Prof Hernani Monteiro,

A B S T R A C T

Anthocyanins are naturally occurring compounds widespread in plant-derived foodstuffs

and therefore abundant in human diet. There are evidences regarding the positive associ-

ation of their intake with healthy biological effects displayed in vivo. This review aims to

highlight some aspects regarding anthocyanins bioavailability; these include a short intro-

ductory part of anthocyanin chemistry, stability, occurrence and intake. This first part is

followed by a more detailed one concerning the main topic of the review that includes

the bioavailability and metabolism of anthocyanins. Special attention is given to the con-

tribution of the gastric mucosa to anthocyanin absorption as the result of the high content

of intact anthocyanins (2025%) detected is plasma few minutes after intake. The contribu-

tion of intestinal tissue and the microbiota impact in anthocyanin absorption and bioactiv-

ity is also highlighted. Despite the biological activities that have been associated with these

compounds, anthocyanins appear to be rapidly absorbed and eliminated, reaching only low

maximal concentrations in plasma and urine. Some possible critical factors that may con-

tribute to this paradox were also explored including the ability of a compound to cross

t of pH, digestive enzymes, biliary acids and microbiota, the lack ofBioavailability of anthocyanins

journal homepage: wwand derivatives

elsevier .com/ locate / j f f4), http://

genetic engineering (Davies, 2009); their benefits for human

health are being discovered, and the applications of anthocy-

anins as colorants or putative bioactives have been exploited

by food, pharmaceutical and cosmetic industries.

2. Anthocyanins chemistry and stability

Attending to their chemical nature, anthocyanins naturally oc-

cur as glycosides of flavylium (2-phenylbenzopyrylium) salts

and are commonly based on six anthocyanidins: pelargonidin

(Pg3glc), cyanidin (Cy3glc), peonidin (Pn3glc), delphinidin

(Dp3glc), petunidin (Pt3glc) and malvidin (Mv3glc) (Fig. 1).

The sugar moieties vary but are usually glucose, rhamnose,

galactose or arabinose (Francis, 1989). The sugar moiety may

be a mono or disaccharide unit, and it may be acylated with

a phenolic or aliphatic acid (Mazza & Miniati, 1993). These

compounds differ in the methoxyl and hydroxyl substitution

patternof the aromatic B ring. Despite themost commonanth-

ocyanidins being just six, there are 539 anthocyanins reported

to be isolated from plants (Andersen & Jordheim, 2005). The

more widespread anthocyanins in fruits are glycosilated in

the 3-OH position (3-O-monoglycosides) and, in less exten-

sion, in both position 3-OH and 5-OH (3,5-O-diglycosides).

Anthocyanins have characteristic physicochemical proper-

ties that confer them its unique colour and stability. They are

between five species (flavylium cation, carbinol base, chal-

cone, quinonoidal base and anionic quinonoidal base) is crit-

ical and deeply related to the colour displayed by

anthocyanins (Fig. 2) (Brouillard & Delaporte, 1977; Brouillard

& Dubois, 1977; Brouillard & Lang, 1990).

3. Occurrence and intake

Polyphenols arise from the secondary metabolism of plants,

being virtually present in all foods and beverages from plant

origin such as vegetables, fruits, cocoa, tea and wine. How-

ever, the daily intake of polyphenols is difficult to estimate

and depend on several factors. Recently, the construction

and application of a database with polyphenols content in

foods has facilitated this task (Neveu et al., 2010).

In 2000, Scalbert and Williamson reported 1 g/day as the

total dietary intake of polyphenols, which were values way

above the ones described for vitamin C or E which are the

classical dietary antioxidants (Scalbert & Williamson, 2000).

Later, Perez-Jimenez and colleagues confirmed polyphenol

daily intake of about 1 g/day by using a French cohort and a

phenolic content in foods database. Even so, this value can

2 J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x xhighly reactive molecules and thus sensitive to degradation

reactions. Oxygen, temperature, light, enzymes and pH are

among the factors that may affect anthocyanins chemistry

and, consequently, their stability and colour. Anthocyanins

may be degraded through several processes occurring during

their extraction, food processing and storage. The fact that in

aqueous solution they co-exist in pH-dependent equilibrium

OR3

O

OH

OH

R1

R2HO

Anthocyanins R1 R2

Pg3glc H H

Pn3glc OCH3 H

Cy3glc OH H

Mv3glc OCH3 OCH3

Pt3glc OCH3 OH

Dp3glc OH OHFig. 1 Representation of the general structure of

anthocyanins (flavylium form). R3 is a sugar moiety.

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010be higher due to a lack or insufficient data on food contents

for more complex polyphenols (Perez-Jimenez et al., 2011).

Anthocyanins are part of human diet as they can be found

in red wine, some cereals and root vegetables (aubergines,

beans, cabbage, radishes, onions) but mainly due to its pres-

ence in red fruits such as cherries, strawberries, plums, black-

berries, raspberries, grapes, red currants and black currants.

Anthocyanins are mainly found in skin but can also appear

in the flesh, e.g., cherries and strawberries. Usually, the con-

tent in anthocyanins increases during ripening and can reach

values up to 24 g/kg fresh weight in blackcurrants, blackel-

derberry or blackchokeberry (Clifford, 2000; Mazza & Velioglu,

1992; Perez-Jimenez, Neveu, Vos, & Scalbert, 2010). Redwine is

Fig. 2 Schematic representation of the molar fraction ofanthocyanin equilibrium form according to the GI tract pH

adapted from (Nave et al., 2010).

thocyanins and derivatives, Journal of Functional Foods (2014), http://

also a good source of anthocyanins and contains approxi-

mately 200350 mg of anthocyanins per L; in addition, as

the wine ages, these anthocyanins can be converted into

more complex structures such as anthocyanin-pyruvic

acid adducts and vinylpyranoanthocyanin-catechins (Fig. 3)

(Clifford, 2000; Oliveira, de Freitas, Silva, & Mateus, 2007;

Perez-Jimenez et al., 2010; Pissarra et al., 2004; Silberberg

et al., 2006; Sousa et al., 2007).

4. Bioavailability and metabolism ofanthocyanins

Anthocyanins, for consumers that eat berries and drink red

wine on a routine basis, are major dietary components. How-

ever, the key difference compared to the other flavonoid gly-

cosides, is that anthocyanins undergo re-arrangements in

response to pH and temperature (Brouillard & Delaporte,

1977) (Fig. 2). Physiological temperatures are highly suitable

both thermodynamically and kinetically for observing the

chalcone tautomer (Brouillard & Delaporte, 1977). The limited

available experimental evidence indicates that in the acidic

conditions that prevail in the gastric compartment anthocya-

nins are in the positively charged flavylium form, whilst all

the other dietary flavonoids remain neutral.

The difficulty in overcoming those analytical problems

may contribute significantly to the low bioavailability of

anthocyanins, which does not justify all the biological activi-

ties previously associated with the huge consumption of this

flavonoid class.

Extensive knowledge of the bioavailability of anthocyanins

is thus essential if their health effects are to be understood

(Fig. 4).

After ingestion, anthocyanins are readily detected in plas-

ma in their parent forms, possibly as a result of their absorp-

tion through the gastric wall (Cao, Muccitelli, Sanchez-

Moreno, & Prior, 2001; Cao & Prior, 1999; Milbury, Cao, Prior,

& Blumberg, 2002; Mulleder, Murkovic, & Pfannhauser, 2002).

It has been only recently that studies were conducted to

determine tissue concentrations of anthocyanins. The stom-

ach exhibited only native anthocyanins, while in other organs

(jejunum, liver, and kidney) native and methylated anthocya-

nins as well as conjugated anthocyanidins (monoglucuro-

nides) were detected (Talavera et al., 2005).

In another work, pigs were fed with diets supplemented

with blueberries for 4 weeks. Although no anthocyanins were

detected in the plasma or urine of the fasted animals, intact

anthocyanins were detected in the liver, eye, cortex, and cer-

ebellum. The results suggest that anthocyanins can accumu-

J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x x 3Fig. 3 Chemical structures of anthocyanin derivatives: malvidi

glucoside-catechin and (+)-catechin-(4,8)-malvidin-3-glucoside.

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010n-3-glucoside pyruvic acid adduct, vinylpyranomalvidin-3-


late in tissues, including tissues beyond the bloodbrain bar-

rier (Kalt et al., 2008).

In a more recent work, anthocyanin metabolites (methyl-

ated anthocyanins and glucurono-conjugated derivatives)

were identified in various organs (bladder, prostate, testes,

heart and adipose tissue) in rats fed with a blackberry antho-

cyanin-enriched diet for 12 days (Felgines et al., 2009). In that

study, the bladder contained the highest levels of anthocya-

nins, followed by the prostate. Prostate, testes and heart con-

tained native cyanidin 3-glucoside and a small proportion of

cyanidinmonoglucuronide. Cyanidin 3-glucoside andmethyl-

ated derivatives were present in adipose tissue. Moreover, two

recent works reported the capacity of dietary anthocyanins

from grapes and berries to reach the brain (Passamonti,

Vrhovsek, Vanzo, & Mattivi, 2005; Talavera et al., 2005).

4.1. Oral cavity absorption of anthocyanins

Studies involving individual anthocyanins revealed that their

amount in plasma is generally 1% of the consumed quanti-

ties, due to limited intestinal absorption, although additional

factors may contribute to the proposed low anthocyanin bio-

availability such as high rates of cellular uptake, metabolism

and excretion (Fig. 5) (Manach, Williamson, Morand, Scalbert,

& Remesy, 2005).

Upstream to gastrointestinal absorption, a variety of bind-

ing processes can take place, namely interaction with food

proteins or with salivary proteins and digestive enzymes

(Matsui et al., 2001; Walle, Browning, Steed, Reed, & Walle,

2005; Wiese, Gartner, Rawel, Winterhalter, & Kulling, 2009).

In a recent work with healthy volunteers, black raspberry

anthocyanins could be detected as their hydrolyzed aglycone

form in the oral cavity, resulting from the activity of b-glyco-

sidase derived both from bacteria and oral epithelial cells

(Mallery et al., 2011). In the same study, parent anthocyanins

and protocatechuic acid, a cyanidin-3-glucoside microbiota

metabolite, were detected in the saliva. Furthermore, saliva

samples revealed the presence of glucuronidated anthocya-

nin conjugates, consistent with intracellular uptake and

phase II conversion of anthocyanins (Mallery et al., 2011).

Still, whether these oral transformation reactions will be

of importance for local effects in the oral epithelium is diffi-

cult to assess considering the relatively short residence time

of most foods in the oral cavity.

4.2. Gastric absorption of anthocyanins a new approach

Until the beginning of the twenty first century the study of

fast kinetics of plasma appearance of anthocyanins in rats

and humans was a challenging field (Cao & Prior, 1999; Mil-

4 J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x xFig. 4 Hypothetic pathways of anthocyanins absorption, distri

information.

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010bution, metabolism and excretion based on current


O Obury et al., 2002; Murkovic, Mulleder, Adam, & Pfannhauser,

2001; Tsuda, Horio, & Osawa, 1999).

In 2003, Passamonti and co-workers performed an in vivo

experiment in rats that suggested the ability of anthocyanins

to cross the gastric mucosa (Passamonti, Vrhovsek, Vanzo, &

Mattivi, 2003).

Later, Talavera et al., 2003 extended their research to struc-

turally related anthocyanins and demonstrated that anthocy-

anin glycosides were quickly and efficiently absorbed in the

stomach (approximately 25%). However their absorption var-

ied greatly according to the anthocyanin structure and they

were rapidly excreted into bile as intact and metabolized

forms.

In 2005, the same authors investigated anthocyanin

metabolism and distribution in different rat organs (stomach,

jejunum, liver, kidney and brain). The only additional infor-

mation of this work concerning the stomach absorption was

its incapacity of metabolizing anthocyanins, since no metab-

olites were detected in this organ.

Moreover, El Mohsen et al. (2006) have analyzed pelargoni-

Isofla

vone

s

Flava

none

s

Flavo

nols

Flava

nols

Phen

olic a

cids

Antho

cyan

ins0

1

2

3

4

Cm

ax (

M)

Fig. 5 Maximum plasma concentration of anthocyanins

compared to other flavonoid classes adapted from (Manach

et al., 2005).

J O U R N A L O F F U N C T I O N A L Fdin gastric absorption in rats having detected the presence of

p-hydroxybenzoic acid in stomach 2 h after ingestion. This re-

sult is not indicative of anthocyanin transformation in gastric

cavity but rather of the instability and degradation of the

anthocyanidin.

The absorption of red orange anthocyanins was studied in

both rat stomach and intestine using in situ models (Felgines

et al., 2006). A high proportion (about 20%) of red orange

anthocyanins was absorbed from the stomach and again no

anthocyanin metabolite was observed in the stomach after

30 min of incubation.

The earliest reference on anthocyanin absorption in stom-

ach is from 2007. In that work the authors examined the gas-

tric absorption of pelargonidin-3-glucoside using rat models.

Once more, a high proportion of pelargonidin-3-glucoside

was found to be rapidly absorbed from the stomach (23%) (Fel-

gines et al., 2007).

Bearing all the above data, it appears that these amazing

compounds could entirely jump the gastric system and

reach the systemic circulation in their native or metabolized

forms, being available to exert their biological activities

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010approximately 30 min after ingestion. In the meanwhile, all

other flavonoid glycoside compounds are still in their journey

towards the circulation. There are some few references con-

cerning the gastric absorption of flavonoid aglycones. Querce-

tin, but not quercetin 3-O-glucoside nor quercetin 3-O-

rutinoside, was found to be absorbed in rat stomach (Crespy

et al., 2001). Similarly, the isoflavones genistein and daidzein,

but not their glucosides, were also found to be absorbed in the

rat stomach (Piskula, Yamakoshi, & Iwai, 1999). Finally, some

phenolic acids were also found to be absorbed at the gastric

epithelium (Konishi, Zhao, & Shimizu, 2006; Lafay et al.,

2006; Vanzo et al., 2007; Zhao, Egashira, & Sanada, 2004).

The main conclusion common to all the authors is the

high content of anthocyanins in the stomach, but the possible

mechanism of anthocyanin gastric absorption remains

unknown.

On this matter, the bioavailability of cyanidin-3-glucoside

in rats that were fed a red orange extract with or without glu-

cose by gastric intubation was not significantly affected by

simultaneous ingestion of glucose (Felgines et al., 2008). This

fact may suggest that the glucose transporters are not in-

volved in anthocyanin gastric absorption.

Information regarding the kinetic flux of anthocyanins in

the GI is critical for understanding anthocyanin absorption.

After feeding rats with black raspberry extract by stomach

tube, anthocyanin content in the gastric lumen was found

to decrease linearly during 180 min (He, Wallace, Keatley, Fail-

la, & Giusti, 2009). The estimated time to deplete half of the

anthocyanin content in the gastric lumen of the fasted rat

was approximately 120 min, suggesting that minimal

amounts of anthocyanins would still be present in the stom-

ach after 4 h. In the same work, the authors highlighted an-

other important factor that may contribute to the

underestimated anthocyanin quantification. Anthocyanins

appeared to bind to unidentified protein in the stomach tissue

and thus could not be quantified as free anthocyanins by

HPLC. Such binding may be attributed to nonspecific binding

or perhaps specific binding to some protein transporter.

The organic anion carrier bilitranslocase is expressed in

the stomach (Battiston, Macagno, Passamonti, Micali, & Luigi

Sottocasa, 1999; Nicolin, Grill, Micali, Narducci, & Passamonti,

2005). Its, in vitro, normal transport activity is competitively

inhibited by quinoidal forms of dietary anthocyanins (Fig. 6),

suggesting that bilitranslocase could promote the facilitated

diffusion of anthocyanins (Passamonti, Vrhovsek, & Mattivi,

2002). Nevertheless, it should be noted that those in vitro as-

says performed were conducted at pH 8.0, which is far from

the gastric conditions here no quinoidal forms could be de-

tected (Fig. 2). Therefore, bilitranslocase may be involved in

anthocyanin quinoidal forms absorption in the liver (Fig. 6).

On the other hand, the administration of high amounts of

anthocyanins, far from diet levels, could induce saturation of

this transport and contribute to the lower anthocyanin bio-

availability reported in those particular studies (Talavera

et al., 2003).

Other transporter candidates may include GLUT1, OAT2,

SMCT1 and SMCT2, since the expression of these transporters

has already been detected in stomach tissue (Eraly, Bush,

D S x x x ( 2 0 1 4 ) x x x x x x 5Sampogna, Bhatnagar, & Nigam, 2004; Garcia, Brown, Pathak,

& Goldstein, 1995; Yoshikawa et al., 2011).


The stomach has been widely ignored as a metabolizing

organ although it has been identified as a site of absorption

for different compounds (Crespy et al., 2001; Piskula et al.,

1999). The contribution of the gastric mucosa to the metabo-

lism of anthocyanins should not be ruled out because the

stomach possesses conjugative enzyme activities (UDP-glu-

curonosyltransferase, sulphotransferase, and catechol-O-

methyl transferase) (Harris, Picton, Singh, & Waring, 2000;

Karhunen, Tilgmann, Ulmanen, Julkunen, & Panula, 1994;

Strassburg, Nguyen, Manns, & Tukey, 1998; Strassburg, Oldha-

fer, Manns, & Tukey, 1997). Besides, in vitro studies showed

that some flavonoids could be metabolized into glucuronidat-

ed and sulphated metabolites by the gastric wall (Dechelotte,

Varrentrapp, Meyer, & Schwenk, 1993; Piskula et al., 1999).

As it can be easily perceived, there is too much ambiguous

information on the literature concerning anthocyanin gastric

absorption.

manner with no statistically differences in their transport

efficiency according to the pH. Attending to the results ob-

tained, a saturable transport for these compounds was thus

proposed.

Considering the previously reported implication of glucose

transporters in the absorption of anthocyanins at the intesti-

nal level (Faria, Pestana, Azevedo et al., 2009) the effect of

anthocyanins on the uptake of 3H-DG was proposed. Preli-

minary studies on this field indicated that the anthocyanins

tested did not affected glucose uptake in MKN-28 cell line

(Fig. 7).

4.3. Intestinal absorption of anthocyanins

The anthocyanin fraction that is not absorbed in the stomach

reaches the small intestine. Once anthocyanins enter more

basic conditions in the small intestine the carbinol pseudo-

1

6 J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x xWorking with animal models or human volunteers is obvi-

ously an important vehicle for obtaining new insights on

anthocyanin bioavailability. In the meantime, a gastric cell

barrier model would be extremely useful similarly to what

was gained with the studies with caco-2 cell line that mimics

the intestinal barrier.

The methods available to evaluate the absorption of drugs

at the gastric level make use of isolated gastric epithelial cells,

which are both time and labour consuming.

A critical feature of such a model is that it has to work in

the presence of a reduced pH. A recently published work re-

ported the development of a biologically relevant in vitro mod-

el of moderately differentiated adenocarcinoma stomach

cells (MKN-28) to be used as a gastric barrier model (Fernan-

des, de Freitas, Reis, & Mateus, 2012). In that work, the

absorption and metabolism of anthocyanins through gastric

epithelium cells was evaluated over time in the presence of

proton gradient.

By using this model, it was possible to study different pH

conditions that correspond to the fed and unfed stage, pH

of 3.0 or 5.0, respectively (Dressman et al., 1990; Russell

et al., 1993). It was also possible to conclude that anthocya-

nins could cross the gastric epithelium in a time dependentFig. 6 Schematic representation of the in vitro studies conducte

absorption.

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010base is likely to predominate. Unlike flavonoids where glyco-

sides are hydrolyzed, anthocyanin glycosides are rapidly and

efficiently absorbed in the small intestine (Miyazawa, Nakag-

awa, Kudo, Muraishi, & Someya, 1999; Talavera et al., 2004;

Tsuda et al., 1999). Furthermore, anthocyanins are quickly

metabolized and appear in the circulation or are excreted into

bile and urine as both intact and metabolized forms (glucu-

ronidated, sulfated or methylated derivatives) (Fig. 4) (Ichi-

yanagi, Shida, Rahman, Hatano, Matsumoto et al., 2005;

Ichiyanagi, Shida, Rahman, Hatano, et al., 2005; Ichiyanagi

et al., 2004; McGhie, Ainge, Barnett, Cooney, & Jensen, 2003;

Miyazawa et al., 1999; Talavera et al., 2003; Talavera et al.,

2004).

The potential mechanisms of anthocyanin glycosides

absorption in the small intestine may involve a specific glu-

cose transporter, such as SGLT1, as previously suggested for

other flavonoids (Hollman et al., 1999).

A recent work points for the putative involvement of

GLUT2 transporter in anthocyanins absorption at the intesti-

nal level (Faria et al., 2009).

Another possible mechanism may involve the hydrolyza-

tion of anthocyanins by brush border enzymes such as lactase

phloridzin hydrolase, prior to passive diffusion of the agly-

2

1

2d to prove the involvement of bilitranslocase in anthocyanin


Dp

ak

pre

0 lM

O Ocone, as already proved for other flavonoids (Gee et al., 2000;

Hollman et al., 1999).

Unabsorbed anthocyanins reach the colon where they un-

dergo substantial structural modifications. Previous studies

have suggested that this is likely due to the spontaneous deg-

radation under physiological conditions (Woodward, Kroon,

Cassidy, & Kay, 2009) or following microbial metabolism. In

fact, colonic microbiota hydrolyses glycosides into aglycones

and degrades them to simple phenolic acids.

According to Vitaglione and co-workers protocatechuic

acid is the major human metabolite of cyanidin-3-glucoside

in humans (Vitaglione et al., 2007). In particular, proto-

catechuic acid accounts for almost 73% of the ingested antho-

cyanins. This metabolite was detected in plasma 2 h after

orange juice ingestion, indicating that it was possibly formed

through chemical degradation at the physiological conditions

of the systemic circulation or in the intestinal mucosa. This

metabolite is recovered in fecal samples, which suggests that

Contr

ol

Cy3g

lc

Mv3g

lc

Contr

ol0

50

100

150

3 H-2

-deo

xi-D

-glu

cose

upt

ake

(% c

ontr

ol)

Fig. 7 Effect of anthocyanins (100 lM) for 30 min on the upt

incubated at 37 C with 3H-DG, for 1 min. MKN-28 cells were(2 mM) and phloridzin (1 mM) and incubated with 3H-DG (10

Significantly different from the respective control (*p < 0.05).

J O U R N A L O F F U N C T I O N A L Fthe gut extensively metabolizes anthocyanins (Riso et al.,

2005).

The metabolism of berry anthocyanins resulting in pheno-

lic acids in humans was recently studied (Nurmi et al., 2009).

The main anthocyanin metabolites detected were homovani-

lic and vanilic acids.

In another fresh study, blueberry anthocyanin absorption

and metabolism in rats was accomplished and the main

metabolite detected in urine was hippuric acid, which may

be produced in liver through a conjugation of glycine with

aromatic phenolic acids (Del Bo` et al., 2009).

Since anthocyanin phenolic acids can be further absorbed

in colon (Williamson & Clifford, 2010) it is possible that they

are additionally metabolized by hepatic cells (Woodward,

Needs, & Kay, 2011). Health benefits associated with anthocy-

anin rich foods may also be explained by a slow and continu-

ous release of phenolic compounds through the gut into the

bloodstream.

Despite their low apparent bioavailability, plasma concen-

trations of anthocyanins appear sufficient to induce changes

in signal transduction and gene expression in vivo (DeFuria

et al., 2009; Karlsen et al., 2007) in a manner that suggests

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010their putative role in physiological functions and health out-

comes. It is known that flavonoids may directly interact with

membrane lipids altering membrane physical properties, li-

gandreceptor interactions, modulate signal transduction,

transport and enzyme activity (Verstraeten, Fraga, & Oteiza,

2010).

Some of the human studies investigating anthocyanin bio-

availability were recently revised (Faria, Fernandes, Mateus, &

Calhau, 2013).

The overall analysis of the biokinetic parameters of those

studies has facilitated some main assumptions in what

anthocyanin bioavailability is concerned. The most important

one is that although there is a considerable variability in the

values for the biokinetic parameters, anthocyanins appear

to be rapidly absorbed and eliminated, reaching low maximal

concentrations in plasma and urine.

Several factors including variations in the dose, anthocya-

nin chemical composition in the different sources, food or

3glc

Phlor

idzin

1 mM

Phlor

etin 2

mM M

Cytoc

alasin

B 50

*

**

e of 3H-DG (100 lM) by MKN-28 cells. MKN-28 cells were

incubated for 30 min with cytochalasin B (50 lM), phloretin

), for 1 min. Each value represents the mean SEM (n = 6).

D S x x x ( 2 0 1 4 ) x x x x x x 7beverage matrix or processing, age and gender of the individ-

uals and the analytical methodology used can have a huge ef-

fect on the bioavailability and metabolism of anthocyanins.

5. Microbiota impact on anthocyaninsavailability and bioactivity

Microbiota has been considered ametabolizing organ with a

role in human metabolism through endo- and xenobiotic

metabolism, vitamin B12 synthesis, carbohydrate breakdown,

between other important functions.

Besides the obvious role of the gut in normal digestive pro-

cesses, the community of microorganisms in the human GI

tract is now being considered as a microbial organ.

The human gut is composed of a bacterial ecosystem of

around 10131014 bacterial cells, despite not being fully de-

scribed. Microorganisms living inside humans are estimated

to be more than 10 times human cells, and the microbiome

represent more than 100 times the human genome (Cani &

Delzenne, 2009). It has been described a key metabolic role

for microbiota on diabetes and obesity morbilities (Ley et al.,

2005).


mas-Barberan, 2009). Bacteroides could be the genera mostly

O Oinvolved in these reactions as they express these enzymes.

Because of this and, based on substrate specificity, anthocya-

nins may be responsible for the prebiotic benefits associated

with red wine ingestion, in particular for Bacteroides (Que-

ipo-Ortuno et al., 2012). This is a new, but growing, concept,

especially taken into consideration that dysbiosis occurs in

the occidental diet pattern and evidence exists that the Bacte-

roides number needs to grow. Studies comparing microbiota

from lean and obese individuals have shown an increase on

Firmicutes genera and a decrease in Bacteroides (about 50%

reduction) in the obese population (Ley et al., 2005).

The structure of the metabolites produced in colon are not

dependent on sugar moieties but on the structural features of

the polyphenols. As already referred, protocatechuic acid has

been reported as the main metabolite, after anthocyanin con-

sumption (Goldberg et al., 2003; Selma et al., 2009; Slimestad

et al., 2007). A recent work confirmed the degradation of

cyanidin-3-glucoside to protocatechuic acid after incubation

with gut bacteria (Hanske et al., 2013). Curiously, it has been

described that protocatechuic acid improves spatial working

memory (Corona, Vauzour, Hercelin, Williams, & Spencer,

2013).

The individual characteristics of microbiota strongly de-

pend on the dietary habits (Cani & Delzenne, 2011), which

in turn influence anthocyanins bioavailability.

6. Factors affecting anthocyaninsbioavailability

The basic kinetic concepts used to study drug actions or phar-

macokinetics are usually applied to study or predict the

movement of other substances in the organism, such as tox-

ins, environmental pollutants or even phytochemicals.

Biokinetic or disposition is the term used to describe the

path of a xenobiotic in the body and is defined as the compos-

ite actions of its absorption, distribution, biotransformation

and elimination. The bioactivity of a substance is directly

dependent on its concentration, making the disposition of

any compound a major contributor to its potential bioactivity.

Therefore, because the disposition of a chemical determines

its concentration at the site of action, the concerted actionThe chemical forms of anthocyanins ingested in the diet

are not the ones that reach microbiota but instead their

respective metabolites that were excreted in the bile and/or

from the enterohepatic circulation. In colon, anthocyanins

are broadly metabolized by bacteria originating more simple

compounds.

Anthocyanins metabolization includes methylation, sulfa-

tion and conjugation with glucuronic acid but also the break

of glycoside linkages and cleavage of the anthocyanin hetero-

cycle (Goldberg, Yan, & Soleas, 2003; Rechner et al., 2004;

Slimestad, Fossen, & Vagen, 2007). For these reactions to take

place, the involvement of beta-D-glucosidases, beta-D-glucu-

ronidases and alfa-L-rhamnosidases that release aglycones

from their glycoside or glucuronidate forms is necessary

(Aura et al., 2005; Gonthier et al., 2003; Selma, Espin, & To-

8 J O U R N A L O F F U N C T I O N A L Fof absorption, distribution and elimination dictates the po-

tential for biological events to occur.

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010The major route of entry of phytochemicals, polyphenols

and, in particular, flavonoids is by oral ingestion, as they are

consumed as part of a normal diet. The bioavailability of

these xenobiotics can be influenced by several factors, simi-

larly to what happens with other compounds ingested orally.

All the way through the absorption, distribution, biotransfor-

mation and elimination processes, the movement through

biological membrane is implied. Nevertheless, the ability of

a compound to cross membranes can be determined by its

physicochemical characteristics such as size, lipid/water sol-

ubility or pKa. Usually, larger, hydrophilic or ionic charged

molecules cannot freely cross membranes and a transporter

must be implicated.

Regarding the gastrointestinal (GI) tract several other fac-

tors may also influence xenobiotics absorption such as pH,

food, digestive enzymes, biliary acids, microbiota and the

motility and permeability of the GI tract. The passage

through GI is extremely important concerning anthocyanins

absorption since these compounds have a complex chemis-

try responsible for their attractive colours but also for their

instability, which will influence all the biokinetics processes,

and thus, the bioactivity. Its structure is pH-sensitive,

becoming unstable at higher pH (McGhie & Walton, 2007).

The most common methodologies to detect anthocyanins

are centered on their colour in a cationic form and are based

on the total conversion of anthocyanins to this form in

acidic medium. However, the conversion of anthocyanins

to a cation form may not be complete leading to an underes-

timated quantification of these compounds (Fernandes et al.,

2012). In addition, it is usually not considered that anthocy-

anins may be metabolized/biotransformed and, thus, the

conversion back to the cationic form after acidification is

no longer a possibility, contributing to this underestimation.

Further, whether the biological effects attributed to anthocy-

anins are due to their cationic form, their hemiacetal

form or a metabolite of one of these two forms is still

uncertain.

Another important factor affecting anthocyanins bioavail-

ability is their possible ingestion as pigments (anthocyanin

derivatives), especially when considering wine consumption

(Fig. 7). A recent work had already indicated that anthocyanin

pyruvic-acid adducts can rapidly reach rat plasma 15 min

after oral administration of 400 mg/kg bw (21.1 nM kg/lmoland 28.8 nM kg/lmol, of malvidin-3-glucoside-pyruvic acidadduct and malvidin-3-glucoside, respectively) (Faria et al.,

2009). The possible absorption of other forms of anthocyanin

derivatives has also been recently explored in a study that

showed that flavanolanthocyanin pigments presented a

higher absorption efficiency in caco-2 cell model than procy-

anidin B3 (Fernandes, Nave, Goncalves, de Freitas, & Mateus,

2012). This work showed that not only anthocyanins and flav-

anols may cross the caco-2 cell barrier model with similar

efficiency, but also dimeric structures containing both antho-

cyanins and flavanols, although with a lower efficiency than

the respective monomers.

Based on the reported studies, a new field of interest that

is often overlooked arises: the anthocyanin absorption as

anthocyanin-derived pigments.

D S x x x ( 2 0 1 4 ) x x x x x xTherefore, the overall anthocyanin bioavailability should

result from the contribution of the amount that crosses all


O OJ O U R N A L O F F U N C T I O N A L Fphysiological barriers in all their possible forms: native, deg-

radation products, metabolites and anthocyanin derivatives

(Fig. 8).

The availability of phenolic compounds can be also depen-

dent on the food matrix where they are inserted. A more lipo-

philic environment may facilitate flavonoids solubilization

and absorption. Additionally, the presence of ethanol can also

be determinant on the extent of anthocyanins absorption

(Faria et al., 2009), promoting their transport across intestinal

epithelia. Another factor to take into consideration is the

interaction between polyphenolic compounds and the other

compounds present during a meal: it is well known the ability

of these compounds to interact with proteins (Bras et al.,

2010; Goncalves, Mateus, & de Freitas, 2010), modifying or

changing their biological function and limiting and/or inter-

fering with both protein and phenolic absorption; moreover,

because the cell does not have specific mechanisms for phen-

olics entry, they use the cell machinery for other substances,

Fig. 8 Schematic representation of the different anthocyanin f

biological effects of these food components.

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010D S x x x ( 2 0 1 4 ) x x x x x x 9interfering with these molecules absorption, e.g., organic cat-

ions, glucose (Faria, Mateus, de Freitas, & Calhau, 2006; Faria

et al., 2009; Keating, Lemos, Goncalves, & Martel, 2008).

Recently, the frequency of the consumption has also been

a point of discussion since cells long-term exposed to antho-

cyanins demonstrated to be more prone to their transport

(Faria et al., 2009). This point is of specifical importance as it

may be the first step to justify dietary recommendations

and emphasizes the importance of a fruit and vegetable-rich

diet.

Different animal and human studies in the past decade

have related anthocyanin-rich foods with health beneficial ef-

fects (Andres-Lacueva et al., 2005; Krikorian et al., 2010). For

this to happen, bioavailability of these compounds is pre-

sumed but whether the effect is due to the native compounds

or its metabolites, which mechanism are involved or which

factors have crucial impact on bioavailability still remains

underexplored.

orms that could contribute to the net bioavailability and


berries and red wine as main sources, they are gastric and

anins may vary considerably between food sources, environ-

10 J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x xmental conditions, as a direct result of sun exposure,

cultivars, etc. This fact is often neglected in studies, probably

justifying the results variability and the, still existing, gap in

knowledge.

The interest in anthocyanins has been driven primarily by

epidemiological studies that have suggested that diets rich in

these phytochemicals are beneficial to human health.

There is a real possibility that some dietary anthocyanins

or derived pigments contribute positively to health and

well-being. Healthy known effects associated with consump-

tion of anthocyanin-rich foods should be attributed to: (i) di-

rect effects of the absorbed parent compounds (or their

metabolites); (ii) indirect effects mediated by non-absorbed

entities that, probably, induce modifications on microbiota

environment and, consequently, on human metabolism or

could act at the membrane border inducing signal transduc-

tion pathways.

The health benefits associated in epidemiologic studies

with the consumption of anthocyanin-rich foods contradict

the apparent low bioavailability of these compounds.

Nevertheless, the biological activity of absorbed parent

compounds, their metabolites and microbial catabolites and

the potential synergy between them could be the answer to

the anthocyanin paradox bioactivity.

More studies should be carried out in what concerns

anthocyanin or derived pigments transport across biological

membranes. There are studies announcing gastric absorption

and neuroprotective effects of anthocyanin-rich foods, but

there is a gap in the knowledge concerning, for example,

anthocyanin transport across gastric or bloodbrain barrier.

On top of this, it is urgent to know dietary factors able to

modulate anthocyanin bioavailability, helping health profes-

sionals to make dietary recommendations. These recommen-

dations will be relevant for a healthy life but, also, to alert

medical doctors as to possible pharmacological interactions

with anthocyanins.

Acknowledgements

This work was supported by FCT (Fundacao para a Ciencia e

Tecnologia) (POCI, FEDER, POPH, QREN) by studentship grants

(SFRH/BPD/75294/2010 and SFRH/BPD/86173/2012) and one

project grant (PTDC/AGR-TEC/2227/2012).

R E F E R E N C E S

Andersen, O. M., & Jordheim, M. (2005). The anthocyanins. In O. M.Andersen & K. R. Markham (Eds.), Flavonoids: Chemistry,biochemistry and applications (pp. 471552). CRC Press.intestinally absorbed, but they could also reach the colon

and undergo microbiota metabolization. Content of anthocy-7. Conclusion

Overall, anthocyanins are found in diverse food sources, withAndres-Lacueva, C., Shukitt-Hale, B., Galli, R. L., Jauregui, O.,Lamuela-Raventos, R. M., & Joseph, J. A. (2005). Anthocyanins

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010in aged blueberry-fed rats are found centrally and mayenhance memory. Nutritional Neuroscience, 8, 111120.

Aura, A. M., Martin-Lopez, P., OLeary, K. A., Williamson, G.,Oksman-Caldentey, K. M., Poutanen, K., et al. (2005). In vitrometabolism of anthocyanins by human gut microflora.European Journal of Nutrition, 44, 133142.

Battiston, L., Macagno, A., Passamonti, S., Micali, F., & LuigiSottocasa, G. (1999). Specific sequence-directed anti-bilitranslocase antibodies as a tool to detect potentiallybilirubin-binding proteins in different tissues of the rat. FEBSLetters, 453, 351355.

Bras, N. F., Goncalves, R., Mateus, N., Fernandes, P. A., Ramos, M.J., & de Freitas, V. (2010). Inhibition of pancreatic elastase bypolyphenolic compounds. Journal of Agricultural and FoodChemistry, 58, 1066810676.

Brouillard, R., & Delaporte, B. (1977). Chemistry of anthocyaninpigments. 2. Kinetic and thermodynamic study of protontransfer, hydration, and tautomeric reactions of malvidin 3-glucoside. Journal of the American Chemical Society, 99,84618468.

Brouillard, R., & Dubois, J. E. (1977). Mechanism of structuraltransformations of anthocyanins in acidic media. Journal of theAmerican Chemical Society, 99, 13591364.

Brouillard, R., & Lang, J. (1990). The hemiacetal-cis-chalconeequilibrium of malvidin, a natural anthocyanin. CanadianJournal of Chemistry-Revue Canadienne De Chimie, 68, 755761.

Cani, P. D., & Delzenne, N. M. (2009). The role of the gut microbiotain energy metabolism and metabolic disease. CurrentPharmaceutical Design, 15, 15461558.

Cani, P. D., & Delzenne, N. M. (2011). The gut microbiome astherapeutic target. Pharmacology & Therapeutics, 130, 202212.

Cao, G., Muccitelli, H. U., Sanchez-Moreno, C., & Prior, R. L. (2001).Anthocyanins are absorbed in glycated forms in elderlywomen: A pharmacokinetic study. The American Journal ofClinical Nutrition, 73, 920926.

Cao, G., & Prior, R. L. (1999). Anthocyanins are detected in humanplasma after oral administration of an elderberry extract.Clinical Chemistry, 45, 574576.

Clifford, M. N. (2000). Anthocyanins nature, occurrence anddietary burden. Journal of the Science of Food and Agriculture, 80,10631072.

Corona, G., Vauzour, D., Hercelin, J., Williams, C. M., & Spencer, J.P. (2013). Phenolic acid intake, delivered via moderatechampagne wine consumption, improves spatial workingmemory via the modulation of hippocampal and corticalprotein expression/activation. Antioxidants and Redox Signaling[Epub ahead of print].

Crespy, V., Morand, C., Besson, C., Manach, C., Demigne, C., &Remesy, C. (2001). Quercetin, but not its glycosides, is absorbedfrom the rat stomach. Journal of Agricultural and Food Chemistry,50, 618621.

Davies, K. (2009). Modifying anthocyanin production in flowers. InK. Gould, K. Davies, & C. Winefield (Eds.), Anthocyanins Biosynthesis, functions, and applications (pp. 4984). Springer.

Dechelotte, P., Varrentrapp, M., Meyer, H. J., & Schwenk, M. (1993).Conjugation of 1-naphthol in human gastric epithelial cells.Gut, 34, 177180.

DeFuria, J., Bennett, G., Strissel, K. J., Perfield, J. W., Milbury, P. E., &Greenberg, A. S. (2009). Dietary blueberry attenuates whole-body insulin resistance in high fat-fed mice by reducingadipocyte death and its inflammatory sequelae. The Journal ofNutrition, 139, 15101516.

Del Bo`, C., Ciappellano, S., Klimis-Zacas, D., Martini, D., Gardana,C., Riso, P., et al. (2009). Anthocyanin absorption, metabolism,and distribution from a wild blueberry-enriched diet(vaccinium angustifolium) is affected by diet duration in the

SpragueDawley rat. Journal of Agricultural and Food Chemistry,58, 24912497.


J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x x 11Deroles, S. (2009). Anthocyanin biosynthesis in plant cell cultures:A potential source of natural colourants. In K. Gould, K.Davies, & C. Winefield (Eds.), Anthocyanins Biosynthesis,functions, and applications (pp. 107169). Springer.

Dressman, J. B., Berardi, R. R., Dermentzoglou, L. C., Russell, T. L.,Schmaltz, S. P., Barnett, J. L., et al. (1990). Uppergastrointestinal (GI) ph in young, healthy men and women.Pharmaceutical Research, 7, 756761.

El Mohsen, M. A., Marks, J., Kuhnle, G., Moore, K., Debnam, E., Srai,S. K., et al. (2006). Absorption, tissue distribution andexcretion of pelargonidin and its metabolites followingoral administration to rats. British Journal of Nutrition, 95,5158.

Eraly, S. A., Bush, K. T., Sampogna, R. V., Bhatnagar, V., & Nigam, S.K. (2004). The molecular pharmacology of organic aniontransporters: From dna to fda? Molecular Pharmacology, 65,479487.

Faria, A., Fernandes, I., Mateus, N., & Calhau, C. (2013).Bioavailability of anthocyanins. In K. Ramawat & J.-M. Merillon(Eds.). Handbook of natural products (Vol. 2). Germany: Springer.

Faria, A., Mateus, N., de Freitas, V., & Calhau, C. (2006). Modulationof MPP+ uptake by procyanidins in Caco-2 cells: Involvementof oxidation/reduction reactions. FEBS Letters, 580, 155160.

Faria, A., Pestana, D., Azevedo, J., Martel, F., Freitas, V., Azevedo,D., et al. (2009). Absorption of anthocyanins through intestinalepithelial cells - Putative involvement of GLUT2. MolecularNutrition & Food Research, 53, 14301437.

Faria, A., Pestana, D., Monteiro, R., Teixeira, D., Azevedo, J., Freitas,V. D., et al. (2009). Bioavailability of anthocyanin-pyruvic acidadducts in rat. In International conference on polyphenols andhealth (pp. 170171). Yorkshire: Leeds.

Felgines, C., Talavera, S., Texier, O., Besson, C., Fogliano, V.,Lamaison, J. L., et al. (2006). Absorption and metabolism of redorange juice anthocyanins in rats. The British Journal ofNutrition, 95, 898904.

Felgines, C., Texier, O., Besson, C., Lyan, B., Lamaison, J.-L., &Scalbert, A. (2007). Strawberry pelargonidin glycosides areexcreted in urine as intact glycosides and glucuronidatedpelargonidin derivatives in rats. British Journal of Nutrition, 98,11261131.

Felgines, C., Texier, O., Besson, C., Vitaglione, P., Lamaison, J.-L.,Fogliano, V., et al. (2008). Influence of glucose on cyanidin 3-glucoside absorption in rats. Molecular Nutrition & FoodResearch, 52, 959964.

Felgines, C., Texier, O., Garcin, P., Besson, C., Lamaison, J.-L., &Scalbert, A. (2009). Tissue distribution of anthocyanins in ratsfed a blackberry anthocyanin-enriched diet. Molecular Nutrition& Food Research, 53, 10981103.

Fernandes, I., de Freitas, V., Reis, C., & Mateus, N. (2012). A newapproach on the gastric absorption of anthocyanins. Food &Function, 3, 508516.

Fernandes, I., Nave, F., Goncalves, R., de Freitas, V., & Mateus, N.(2012). On the bioavailability of flavanols and anthocyanins:Flavanolanthocyanin dimers. Food Chemistry, 135, 812818.

Francis, F. J. (1989). Food colorants: Anthocyanins. Critical Reviewsin Food Science and Nutrition, 28, 273314.

Garcia, C. K., Brown, M. S., Pathak, R. K., & Goldstein, J. L. (1995).CDNA Cloning of MCT2, a Second MonocarboxylateTransporter Expressed in Different Cells than MCT1. Journal ofBiological Chemistry, 270, 18431849.

Gee, J. M., DuPont, M. S., Day, A. J., Plumb, G. W., Williamson, G., &Johnson, I. T. (2000). Intestinal transport of quercetinglycosides in rats involves both deglycosylation andinteraction with the hexose transport pathway. The Journal ofNutrition, 130, 27652771.

Goldberg, D. M., Yan, J., & Soleas, G. J. (2003). Absorption of three

wine-related polyphenols in three different matrices byhealthy subjects. Clinical Biochemistry, 36, 7987.

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010Goncalves, R., Mateus, N., & de Freitas, V. (2010). Study of theinteraction of pancreatic lipase with procyanidins by opticaland enzymatic methods. Journal of Agricultural and FoodChemistry, 58, 1190111906.

Gonthier, M. P., Cheynier, V., Donovan, J. L., Manach, C., Morand,C., Mila, I., et al. (2003). Microbial aromatic acid metabolitesformed in the gut account for a major fraction of thepolyphenols excreted in urine of rats fed red winepolyphenols. The Journal of Nutrition, 133, 461467.

Hanske, L., Engst, W., Loh, G., Sczesny, S., Blaut, M., & Braune, A.(2013). Contribution of gut bacteria to the metabolism ofcyanidin 3-glucoside in humanmicrobiota-associated rats. TheBritish Journal of Nutrition, 109, 14331441.

Harris, R. M., Picton, R., Singh, S., &Waring, R. H. (2000). Activity ofphenolsulfotransferases in the human gastrointestinal tract.Life Sciences, 67, 20512057.

He, J., Wallace, T. C., Keatley, K. E., Failla, M. L., & Giusti, M. M.(2009). Stability of black raspberry anthocyanins in thedigestive tract lumen and transport efficiency into gastric andsmall intestinal tissues in the rat. Journal of Agricultural andFood Chemistry, 57, 31413148.

Hollman, P. C., Bijsman, M. N., van Gameren, Y., Cnossen, E. P., deVries, J. H., & Katan, M. B. (1999). The sugar moiety is a majordeterminant of the absorption of dietary flavonoid glycosidesin man. Free Radical Research, 31, 569573.

Ichiyanagi, T., Rahman, M. M., Kashiwada, Y., Ikeshiro, Y., Shida,Y., Hatano, Y., et al. (2004). Absorption and metabolism ofdelphinidin 3-O-b-glucopyranoside in rats. Free Radical Biologyand Medicine, 36, 930937.

Ichiyanagi, T., Shida, Y., Rahman, M. M., Hatano, Y., et al. (2005).Extended glucuronidation is another major path of cyanidin 3-O-b-D-glucopyranoside metabolism in rats. Journal ofAgricultural and Food Chemistry, 53, 73127319.

Ichiyanagi, T., Shida, Y., Rahman, M. M., Hatano, Y., Matsumoto,H., Hirayama, M., & Konishi, T. (2005). Metabolic pathway ofcyanidin 3-O-b-D-glucopyranoside in rats. Journal ofAgricultural and Food Chemistry, 53, 145150.

Kalt, W., Blumberg, J. B., McDonald, J. E., Vinqvist-Tymchuk, M. R.,Fillmore, S. A. E., Graf, B. A., et al. (2008). Identification ofanthocyanins in the liver, eye, and brain of blueberry-fed pigs.Journal of Agricultural and Food Chemistry, 56, 705712.

Karhunen, T., Tilgmann, C., Ulmanen, I., Julkunen, I., & Panula, P.(1994). Distribution of catechol-O-methyltransferase enzymein rat tissues. Journal of Histochemistry & Cytochemistry, 42,10791090.

Karlsen, A., Retterstl, L., Laake, P., Paur, I., Kjlsrud-Bhn, S.,Sandvik, L., et al. (2007). Anthocyanins inhibit nuclear factor-kappaB activation in monocytes and reduce plasmaconcentrations of pro-inflammatory mediators in healthyadults. The Journal of Nutrition, 137, 19511954.

Keating, E., Lemos, C., Goncalves, P., & Martel, F. (2008). Acute andchronic effects of some dietary bioactive compounds on folicacid uptake and on the expression of folic acid transporters bythe human trophoblast cell line BeWo. The Journal of NutritionalBiochemistry, 19, 91100.

Konishi, Y., Zhao, Z., & Shimizu, M. (2006). Phenolic acids areabsorbed from the rat stomach with different absorptionrates. Journal of Agricultural and Food Chemistry, 54, 75397543.

Krikorian, R., Shidler, M. D., Nash, T. A., Kalt, W., Vinqvist-Tymchuk, M. R., Shukitt-Hale, B., et al. (2010). Blueberrysupplementation improves memory in older adults. Journal ofAgricultural and Food Chemistry, 58, 39964000.

Lafay, S., Gil-Izquierdo, A., Manach, C., Morand, C., Besson, C., &Scalbert, A. (2006). Chlorogenic acid is absorbed in its intactform in the stomach of rats. The Journal of Nutrition, 136,11921197.Ley, R. E., Backhed, F., Turnbaugh, P., Lozupone, C. A., Knight, R. D.,& Gordon, J. I. (2005). Obesity alters gut microbial ecology.


12 J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x xProceedings of the National Academy of Sciences of the United Statesof America, 102, 1107011075.

Mallery, S. R., Budendorf, D. E., Larsen, M. P., Pei, P., Tong, M.,Holpuch, A. S., et al. (2011). Effects of human oral mucosal tissue,saliva and oral microflora on intraoral metabolism and bioactivationof black raspberry anthocyanins. Cancer Prevention Research.

Manach, C., Williamson, G., Morand, C., Scalbert, A., & Remesy, C.(2005). Bioavailability and bioefficacy of polyphenols inhumans. I. Review of 97 bioavailability studies. The AmericanJournal of Clinical Nutrition, 81, 230242.

Matsui, T., Ueda, T., Oki, T., Sugita, K., Terahara, N., & Matsumoto,K. (2001). a-Glucosidase inhibitory action of natural acylatedanthocyanins. 1. Survey of natural pigments with potentinhibitory activity. Journal of Agricultural and Food Chemistry, 49,19481951.

Mazza, G., & Miniati, E. (1993). Anthocyanins in fruits, vegetables,and grains. In Anthocyanins in fruits, vegetables, and grains. BocaRaton: CRC Press.

Mazza, G., & Velioglu, Y. S. (1992). Anthocyanins and otherphenolic-compounds in fruits of red-flesd apples. FoodChemistry, 43, 113117.

McGhie, T. K., Ainge, G. D., Barnett, L. E., Cooney, J. M., & Jensen, D.J. (2003). Anthocyanin glycosides from berry fruit are absorbedand excreted unmetabolized by both humans and rats. Journalof Agricultural and Food Chemistry, 51, 45394548.

McGhie, T. K., & Walton, M. C. (2007). The bioavailability andabsorption of anthocyanins: Towards a better understanding.Molecular Nutrition & Food Research, 51, 702713.

Milbury, P. E., Cao, G., Prior, R. L., & Blumberg, J. (2002).Bioavailablility of elderberry anthocyanins. Mechanisms ofAgeing and Development, 123, 9971006.

Miyazawa, T., Nakagawa, K., Kudo, M., Muraishi, K., & Someya, K.(1999). Direct intestinal absorption of red fruit anthocyanins,cyanidin-3-glucoside and cyanidin-3,5-diglucoside, into ratsand humans. Journal of Agricultural and Food Chemistry, 47,10831091.

Mulleder, U., Murkovic, M., & Pfannhauser, W. (2002). Urinaryexcretion of cyanidin glycosides. Journal of Biochemical andBiophysical Methods, 53, 6166.

Murkovic, M., Mulleder, U., Adam, U., & Pfannhauser, W. (2001).Detection of anthocyanins from elderberry juice in humanurine. Journal of the Science of Food and Agriculture, 81, 934937.

Nave, F., Petrov, V., Pina, F. R., Teixeira, N., Mateus, N., & de Freitas,V. (2010). Thermodynamic and kinetic properties of a red winepigment: Catechin-(4,8)-malvidin-3-O-glucoside. The Journal ofPhysical Chemistry B, 114, 1348713496.

Neveu, V., Perez-Jimenez, J., Vos, F., Crespy, V., du Chaffaut, L.,Mennen, L., et al. (2010). Phenol-explorer: An onlinecomprehensive database on polyphenol contents in foods. Oxford:Database [bap024].

Nicolin, V., Grill, V., Micali, F., Narducci, P., & Passamonti, S. (2005).Immunolocalisation of bilitranslocase in mucosecretory andparietal cells of the rat gastric mucosa. Journal of MolecularHistology, 36, 4550.

Nurmi, T., Mursu, J., Heinonen, M., Nurmi, A., Hiltunen, R., &Voutilainen, S. (2009). Metabolism of berry anthocyanins tophenolic acids in humans. Journal of Agricultural and FoodChemistry, 57, 22742281.

Oliveira, J., de Freitas, V., Silva, A. M. S., & Mateus, N. (2007).Reaction between hydroxycinnamic acids and anthocyanin-pyruvic acid adducts yielding new portisins. Journal ofAgricultural and Food Chemistry, 55, 63496356.

Passamonti, S., Vrhovsek, U., & Mattivi, F. (2002). The interactionof anthocyanins with bilitranslocase. Biochemical andBiophysical Research Communications, 296, 631636.

Passamonti, S., Vrhovsek, U., Vanzo, A., & Mattivi, F. (2003). The

stomach as a site for anthocyanins absorption from food. FEBSLetters, 544, 210213.

Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010Passamonti, S., Vrhovsek, U., Vanzo, A., & Mattivi, F. (2005). Fastaccess of some grape pigments to the brain. Journal ofAgricultural and Food Chemistry, 53, 70297034.

Perez-Jimenez, J., Fezeu, L., Touvier, M., Arnault, N., Manach, C., &Hercberg, S. (2011). Dietary intake of 337 polyphenols in Frenchadults. The American Journal of Clinical Nutrition, 93, 12201228.

Perez-Jimenez, J., Neveu, V., Vos, F., & Scalbert, A. (2010).Systematic analysis of the content of 502 polyphenols in 452foods and beverages: An application of the phenol-explorerdatabase. Journal of Agricultural and Food Chemistry, 58,49594969.

Piskula, M. K., Yamakoshi, J., & Iwai, Y. (1999). Daidzein andgenistein but not their glucosides are absorbed from the ratstomach. FEBS Letters, 447, 287291.

Pissarra, J., Lourenco, S., Gonzalez-Paramas, A. M., Mateus, N.,Buelga, C. S., Silva, A. M. S., et al. (2004). Structuralcharacterization of new malvidin 3-glucoside-catechin aryl/alkyl-linked pigments. Journal of Agricultural and Food Chemistry,52, 55195526.

Queipo-Ortuno, M. I., Boto-Ordonez, M., Murri, M., Gomez-Zumaquero, J. M., Clemente-Postigo, M., Estruch, R., et al.(2012). Influence of red wine polyphenols and ethanol on thegut microbiota ecology and biochemical biomarkers. TheAmerican Journal of Clinical Nutrition, 95, 13231334.

Rechner, A. R., Smith, M. A., Kuhnle, G., Gibson, G. R., Debnam, E.S., Srai, S. K., et al. (2004). Colonic metabolism of dietarypolyphenols: Influence of structure on microbial fermentationproducts. Free Radical Biology & Medicine, 36, 212225.

Riso, P., Visioli, F., Gardana, C., Grande, S., Brusamolino, A.,Galvano, F., et al. (2005). Effects of blood orange juice intake onantioxidant bioavailability and on different markers related tooxidative stress. Journal of Agricultural and Food Chemistry, 53,941947.

Russell, T. L., Berardi, R. R., Barnett, J. L., Dermentzoglou, L. C.,Jarvenpaa, K. M., Schmaltz, S. P., et al. (1993). Uppergastrointestinal pH in seventy-nine healthy, elderly, NorthAmerican men and women. Pharmaceutical Research, 10,187196.

Scalbert, A., & Williamson, G. (2000). Dietary intake andbioavailability of polyphenols. The Journal of Nutrition, 130,2073S2085S.

Selma, M. V., Espin, J. C., & Tomas-Barberan, F. A. (2009).Interaction between phenolics and gut microbiota: Role inhuman health. Journal of Agricultural and Food Chemistry, 57,64856501.

Silberberg, M., Morand, C., Mathevon, T., Besson, C., Manach, C.,Scalbert, A., et al. (2006). The bioavailability of polyphenols ishighly governed by the capacity of the intestine and of theliver to secrete conjugated metabolites. European Journal ofNutrition, 45, 8896.

Slimestad, R., Fossen, T., & Vagen, I. M. (2007). Onions: A source ofunique dietary flavonoids. Journal of Agricultural and FoodChemistry, 55, 1006710080.

Sousa, C., Mateus, N., Silva, A. M. S., Gonzalez-Paramas, A. M.,Santos-Buelga, C., & de Freitas, V. (2007). Structural andchromatic characterization of a new malvidin 3-glucoside-vanillyl-catechin pigment. Food Chemistry, 102, 13441351.

Strassburg, C. P., Nguyen, N., Manns, M. P., & Tukey, R. H. (1998).Polymorphic expression of the UDP-glucuronosyltransferaseUGT1A gene locus in human gastric epithelium. MolecularPharmacology, 54, 647654.

Strassburg, C. P., Oldhafer, K., Manns, M. P., & Tukey, R. H. (1997).Differential expression of the UGT1A locus in human liver,biliary, and gastric tissue: Identification of UGT1A7 andUGT1A10 transcripts in extrahepatic tissue. MolecularPharmacology, 52, 212220.Talavera, S., Felgines, C., Texier, O., Besson, C., Gil-Izquierdo, A.,Lamaison, J. L., et al. (2005). Anthocyanin metabolism in rats


and their distribution to digestive area, kidney, and brain.Journal of Agricultural and Food Chemistry, 53, 39023908.

Talavera, S., Felgines, C., Texier, O., Besson, C., Lamaison, J. L.,et al. (2003). Anthocyanins are efficiently absorbed from thestomach in anesthetized rats. The Journal of Nutrition, 133,41784182.

Talavera, S., Felgines, C., Texier, O., Besson, C., Manach, C.,Lamaison, J. L., & Remesy, C. (2004). Anthocyanins areefficiently absorbed from the small intestine in rats. TheJournal of Nutrition, 134, 22752279.

Tsuda, T., Horio, F., & Osawa, T. (1999). Absorption andmetabolism of cyanidin 3-O-b-D-glucoside in rats. FEBS Letters,449, 179182.

Vanzo, A., Cecotti, R., Vrhovsek, U., Torres, A. M., Mattivi, F., &Passamonti, S. (2007). The fate of trans-caftaric acidadministered into the rat stomach. Journal of Agricultural andFood Chemistry, 55, 16041611.

Verstraeten, S., Fraga, C., & Oteiza, P. (2010). Flavonoidsmembrane interactions: Consequences for biological actions.In C. Fraga (Ed.), Plant phenolics and human health (pp. 107136).John Wiley & Sons, Inc..

Vitaglione, P., Donnarumma, G., Napolitano, A., Galvano, F., Gallo,A., Scalfi, L., et al. (2007). Protocatechuic acid is the majorhuman metabolite of cyanidin-glucosides. The Journal ofNutrition, 137, 20432048.

Walle, T., Browning, A. M., Steed, L. L., Reed, S. G., & Walle, U. K.(2005). Flavonoid glucosides are hydrolyzed and thus

activated in the oral cavity in humans. The Journal of Nutrition,135, 4852.

Wiese, S., Gartner, S., Rawel, H. M., Winterhalter, P., & Kulling, S. E.(2009). Protein interactions with cyanidin-3-glucoside and itsinfluence on a-amylase activity. Journal of the Science of Food andAgriculture, 89, 3340.

Williamson, G., & Clifford, M. N. (2010). Colonic metabolites ofberry polyphenols: The missing link to biological activity?British Journal of Nutrition, 104, S48S66.

Woodward, G., Kroon, P., Cassidy, A., & Kay, C. (2009). Anthocyaninstability and recovery: Implications for the analysis of clinicaland experimental samples. Journal of Agricultural and FoodChemistry, 57, 52715278.

Woodward, G. M., Needs, P. W., & Kay, C. D. (2011). Anthocyanin-derived phenolic acids form glucuronides following simulatedgastrointestinal digestion and microsomal glucuronidation.Molecular Nutrition & Food Research, 55, 378386.

Yoshikawa, T., Inoue, R., Matsumoto, M., Yajima, T., Ushida, K., &Iwanaga, T. (2011). Comparative expression of hexosetransporters (SGLT1, GLUT1, GLUT2 and GLUT5) throughoutthe mouse gastrointestinal tract. Histochemistry and Cell Biology,135, 183194.

Zhao, Z., Egashira, Y., & Sanada, H. (2004). Ferulic acid is quicklyabsorbed from rat stomach as the free form and thenconjugated mainly in liver. The Journal of Nutrition, 134,30833088.

J O U R N A L O F F U N C T I O N A L F O O D S x x x ( 2 0 1 4 ) x x x x x x 13Please cite this article in press as: Fernandes, I. et al., Bioavailability of andx.doi.org/10.1016/j.j.2013.05.010thocyanins and derivatives, Journal of Functional Foods (2014), http://

Bioavailability of anthocyanins and derivatives1 Introduction2 Anthocyanins chemistry and stability3 Occurrence and intake4 Bioavailability and metabolism of anthocyanins4.1 Oral cavity absorption of anthocyanins4.2 Gastric absorption of anthocyanins a new approach4.3 Intestinal absorption of anthocyanins

5 Microbiota impact on anthocyanins availability and bioactivity6 Factors affecting anthocyanins bioavailability7 ConclusionAcknowledgementsReferences

bioavailability of anthocyanins and derivatives

Documents

x x x x xavai lab

university of porto

f o o d s x x x

anthocyanins bioavailability

portugalrsity of porto

o f f u n c t i o n

portugala r t i c

byj o u r n