Handbook of Anthocyanins. Anthocyanins: Food sources and benefits to consumer´s health

Abstract

Anthocyanins are one of the most abundant flavonoid compounds. These pigments, naturally present in fruits and vegetables, provide color and promote health benefits to consumers due to their antioxidant capacity. To date, more than 600 anthocyanins have been identified in nature, all coming from six anthocyanidin aglycones derived from flavylium backbone with different glycosylations and acylations. The different anthocyanin conjugates absorb light at about 500 nm and are responsible for the red, blue and purple color of fruits and vegetables. Many studies in cell lines, animal models and human clinical trials suggest that anthocyanins have anti- carcinogenic and anti- inflammatory activities, provides cardiovascular disease prevention, promote obesity and diabetes control benefits, and also improve visual and brain functions. Those health benefits are mainly associated with their antioxidant effects, which clearly are influenced by the molecular mechanism related to the expression and modulation of key genes. The bioavailability of anthocyanins in functional foods is one of the questions to solve regarding their putative health-promoting effects... Is there any correlation between food sources with more anthocyanins content and their higher health benefits? What is the best source of anthocyanins to obtain the highest health-promoting properties? These are questions covered in the present article.

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Chapter
ANTHOCYANINS: FOOD SOURCES AND BENEFITS
TO CONSUMER’S HEALTH
Patricio Ramos, Raul Herrera and María Alejandra Moya-León
*
Laboratorio de Fisiología Vegetal y Genética Molecular, Instituto de Ciencias Biológicas,
Universidad de Talca, 2 Norte 685, Talca, Chile.
A
BSTRACT
Anthocyanins are one of the most abundant flavonoid compounds. These pigments,
naturally present in fruits and vegetables, provide color and promote health benefits to
consumers due to their antioxidant capacity. To date, more than 600 anthocyanins have
been identified in nature, all coming from six anthocyanidin aglycones derived from
flavylium backbone with different glycosylations and acylations. The different
anthocyanin conjugates absorb light at about 500 nm and are responsible for the red, blue
and purple color of fruits and vegetables. Many studies in cell lines, animal models and
human clinical trials suggest that anthocyanins have anti- carcinogenic and anti-
inflammatory activities, provides cardiovascular disease prevention, promote obesity and
diabetes control benefits, and also improve visual and brain functions. Those health
benefits are mainly associated with their antioxidant effects, which clearly are influenced
by the molecular mechanism related to the expression and modulation of key genes. The
bioavailability of anthocyanins in functional foods is one of the questions to solve
regarding their putative health-promoting effects... Is there any correlation between food
sources with more anthocyanins content and their higher health benefits? What is the best
source of anthocyanins to obtain the highest health-promoting properties? These are
questions covered in the present article.
I
NTRODUCTION
During the last years increasing consideration has been placed in plants and foods which
can contain antioxidant properties. This particular attention in fruits and vegetables is due to
multiple reports indicating their effectiveness in the reduction of risks of chronic diseases in
*
Corresponding author: Email: alemoya@utalca.cl.

Patricio Ramos, Raul Herrera and María Alejandra Moya-León
2
humans (Steinberg, 1995; Olsson et al., 2004; Yi et al., 2006; Hassan and Abdel-Aziz, 2010).
The chemical compounds present in vegetables and fruits that are related to health-promoting
benefits considered several antioxidants, as vitamins C and E, carotenoids and flavonoids; the
last ones are the most diverse group with over 9,000 structural variants known (Williams and
Grayer, 2004). The chemical variety, molecular weight, three-dimensional conformation,
biochemical and physical properties of these molecules allow them to interact with different
targets in many live organisms (Taylor and Grotewold, 2005; Peer and Murphy, 2007).
Pigmented flavonoids, mainly anthocyanins, are considered the most important group of
flavonoids in plants having more than 600 compounds identified in nature (Andersen and
Jordheim, 2008). Anthocyanins are water-soluble compounds that provide color to plant
tissues (leaves, stems, roots, flowers and fruits) ranging from red, purple to blue according to
the environmental pH and their structural composition (Fossen et al., 1996). In fact, the word
anthocyanin is derived from the Greek words 'anthos' meaning flower and 'kyanos' dark blue.
Regarding to human consumption, the high intake of foods rich in anthocyanins offers
potential health beneficial effects on various disorders associated with cancer, aging diseases,
obesity, neurological diseases, inflammation, diabetes as well as bacterial infections (Hudson
et al., 2000; Olsson et al., 2004; Yi et al., 2006; De Pascual-Teresa and Sanchez-Ballesta,
2008; Hassan and Abdel-Aziz, 2010; Sancho and Pastore, 2012). Here, we review the
information available concerning the structure, composition and abundance of anthocyanins
in fruits, its bioavailability and biological activity related to health promoting effects, with
special emphasis in strawberry fruit.
C
HEMICAL
S
TRUCTURE
Anthocyanins belong to the widespread class of polyphenolic compounds named
flavonoids that are synthesized by plants as part of their secondary metabolism. Based on
their chemical characteristics, flavonoids are divided into different subclasses: flavonols,
flavanols, anthocyanidins, flavanones, flavones and isoflavones. They share a common
carbon skeleton denominated flavylium ion backbone, consisting in C-6 (A ring)-C-3 (C
ring)-C-6 (B ring) (Figure 1) (Harborne, 1998). The differences between the subclasses are
due to the existence of double bonds and carbonyl groups in the C ring.
Anthocyanins are glycosylated anthocyanidins with associated in their structure (Figure
1), which are the most common type of pigments found in plants. In nature, it is possible to
find hundreds of different anthocyanins due to complex glycosylation patterns over a few
aglycones (Table 1). In fact, there are around 25 different aglycones identified in nature to
date. Nevertheless, only six anthocyanidins (cyanidin, delphinidin, malvidin, pelargonidin,
peonidin, and petunidin) are widely distributed in nature, accounting for over 90% of the
anthocyanins identified (Eder, 2000; Kong et al., 2003; Andersen and Jordheim, 2006).
Anthocyanins are intensely colored at low pH conditions due to the presence of eight
conjugated double bonds giving a positive charge to the molecule. Color variation is due to
the differential substitutions in B ring of the backbone, being blue with hydroxyl substitutions
and red with methoxyl groups (Heredia et al., 1998; Delgado-Vargas and Paredes-Lopez,
2003). These substitutions affect the physical and chemical properties of anthocyanins
modifying their molecular size and polarity. Glycosylation increases water solubility, whereas

Anthocyanins: Food Sources and Benefits to Consumer’s Health 3
acylation decreases it. Sugar addition to the aglycones improves anthocyanin stability
favoring the formation of intramolecular H-bonding network within the molecule (Borkowski
et al., 2005). Glucose (glu) and rhamnose (rha) are the most frequent sugars attached to the
aglycone, but galactose (gal), arabinose (ara), xylose (xyl), rutinose (rut) are also added. In
addition to glycosylations, it is possible to find acylations by aliphatic and cinnamic acids. All
these substitutions in the anthocyanin backbones provide a wide range of molecules in nature.
D
ISTRIBUTION AND
A
BUNDANCE IN
P
LANTS
Anthocyanins are widely distributed in nature and present in different plant organs,
including flowers, fruits, roots, leaves and stems (Brouillard, 1982; Delgado-Vargas and
Paredes-Lopez, 2003; Shahidi and Naczk, 2004). Due to their chemical structure
anthocyanins are soluble in water solutions and frequently found in the vacuoles of epidermal
cells, and in some species, they are bound to membrane of the main cell-vacuole, called
anthocyanoplasts (Pecket and Small, 1980).
The flavylium ion backbone, consisting in C-6 (A ring)-C-3 (C ring)-C-6 (B ring) is the common
carbon skeleton from anthocyanidin aglycones. The most abundant anthocyanidins in nature are
cyanidin (Cy), delphinidin (Dp), malvidin (Mv), pelargonidin (Pg), peonidin (Pn), and petunidin
(Pt), and they share the following substitution patterns in the common skeleton backbone: OH in
R1, OH in R2, H in R3, OH in R4, OH in R6. Nevertheless, differential substitutions are found in
R5 (OH: Cy, Dp; OMe: Mv, Pn, Pt; H, Pg) and R7 (OH: Dp, Pt; OMe: Mv; H: Cy, Pg, Pn).
Figure 1. General structure of anthocyanidin aglycones.
Table 1. Structural information of the most abundant anthocyanidin aglycones found in
nature. Substitution patterns in the common skeleton backbone described in Figure 1
(Adapted from Casteñeda-Ovando et al., 2009)
Substitution pattern
Name Abbreviations R1 R2 R3 R4 R5 R6 R7 Colour
Cyanidin Cy OH OH H OH OH OH H Orange-red
Delphinidin Dp OH OH H OH OH OH OH Blue-red
Malvidin Mv OH OH H OH OMe OH OMe Blue-red
Pelargonidin Pg OH OH H OH H OH H Orange-red
Peonidin Pn OH OH H OH OMe OH H Orange-red
Petunidin Pt OH OH H OH OMe OH OH Blue-red

Patricio Ramos, Raul Herrera and María Alejandra Moya-León
4
Table 2. Concentration of anthocyanins in fruits and fruit sub-products.
(Adapted from De Pascual-Teresa and Sanchez-Ballesta, 2008)
Fruit/sub-product Anthocyanin (mg/100g)
Apple, Red Delicious 1.7
Bilberry 300-698
Black currant 130-476
Black olives 42-228
Blackberry 82.5-325.9
Blueberry 25-495
Cherry 2-450
Chokeberry 410-1480
Cranberry 67-140
Crowberry 360
Elderberry 200-1816
Gooseberry 2-43.3
Peach 4.2
Pomegranate (juice) 600-765
Port wine 14-110
Purple corn 1642
Raspberry 20-687
Red apple 1.3-12
Red grape 30-750
Red wine 16.4 - 35
Saskatoon berry 234
Strawberry 19-55
The relative abundance of anthocyanins may vary according to the species or from fruit
to fruit of the same species depending on external and internal factors. Genetic factors and
agronomic practices, intensity and type of light, temperature, processing and storage
conditions influence the level of anthocyanins.
Extensive research has been carried out in order to identify the major plant food sources
of anthocyanins (Wu et al., 2006; Ogawa et al., 2008; Li et al., 2012). Table 2 listed the main
sources of anthocyanin from fruit species and sub-products (De Pascual and Sanchez-
Ballesta, 2008).
Among the most common anthocyanidins in higher plants, the glycosides of the three
non-methylated anthocyanidins (cyanidin, delphinidin and pelargonidin) are the most
abundant in nature, representing 80% of leaf pigments, 69% of fruits and 50% of flowers. The
distribution of the six most common anthocyanidins in the edible parts of plants is cyanidin
(50%), pelargonidin (12%), peonidin (12%), delphinidin (12%), petunidin (7%) and malvidin
(7%). The most widespread anthocyanin in fruits is cyanidin-3-glucoside (Kong et al., 2003).
Red and blue highly pigmented fruits, mainly berries such as blueberry, blackberry,
blackcurrant, cherry, cranberry, raspberry and strawberry fruits, have been comprehensively
analyzed suggesting that anthocyanins contribute significantly to the antioxidant activity
(Yang et al., 2008; Juroszek et al., 2009). Recently, anthocyanin composition and its
antioxidant capacity were determined for a list of highly pigmented edible vegetables (Li et
al., 2012). Consistently, fruits show the highest level of anthocyanins, which is indicative of

Anthocyanins: Food Sources and Benefits to Consumer’s Health 5
the best natural source of these compounds. The vegetables with higher cyanidin content and
with superior total anthocyanin level show better antioxidant activities.
Berries are widely consumed fruits, not only in fresh and frozen forms, but also as
processed and derived products, including dried and canned fruits, yogurts, beverages, jams,
and jellies (Seeram, 2006). In the specific case of strawberry fruit, anthocyanins are the most
studied phenolic compounds and the principal in terms of quantity (Clifford, 2000; Lopes da
Silva et al., 2002) (Table 3). Many studies have determined total anthocyanin content,
reporting values ranging from 150 to 800 mg/kg of fresh weight (Garcia-Viguera et al., 1998;
Clifford, 2000; Castro et al., 2002). Pelargonidin 3-glucoside is the predominant anthocyanin
in several varieties of strawberries, followed by pelargonidin 3-rutinoside and cyanidin 3-
glucoside (Gil et al., 1997; Kosar et al., 2004; Tulipani et al., 2008). These three compounds
represent more than 95% of the total content of anthocyanins in the commercial strawberry
(Lopes da Silva et al., 2007).
Table 3. Flavonoids composition reported in strawberries.
(Adapted from Giampieri et al. 2012)
Group Compound
Anthocyanins Cyanidin-3-glucoside
Cyanidin-3-rutinoside
Cyanidin-3-malonylglucoside
Cyanidin-3-malonylglucosyl-5-glucoside
Pelargonidin-3-galactoside
Pelargonidin-3- glucoside
Pelargonidin-3- rutinoside
Pelargonidin-3- arabinoside
Pelargonidin-3,5- diglucoside
Pelargonidin-3- malylglucoside
Pelargonidin-3- malonylglucoside
Pelargonidin-3- acetylglucoside
Pelargonidin-disacaride (hexose + pentose) acylated with acetic acid
5-pyranopelargonidin-3-glucoside
Flavonols Quercetin-3- glucuronide
Quercetin -3- malonylglucoside
Quercetin - rutinose
Quercetin - glucoside
Quercetin - glucoronide
Kaempferol -3- glucoside
Kaempferol -3- malonylglucoside
Kaempferol - coumaroyl - glucoside
Kaempferol - glucoronide
Flavanols Proanthocyanidin B1(EC-4,8-C)
Proanthocyanidin trimer (EC-4,8-EC-4,8-C)
Proanthocyanidin B3 (C-4,8-C)
(+) - Catechin

Patricio Ramos, Raul Herrera and María Alejandra Moya-León
6
Commercial strawberry (Fragaria x ananassa Duch. ex Rozier) was originated from an
accidental cross of the white-fruited Chilean strawberry (F. chiloensis (L.) Mill. ssp.
chiloensis f. chiloensis) and the meadow strawberry (F. virginiana Mill. ssp. virginiana) that
occurred in a Royal Botanical Garden in France (Darrow, 1966; Staudt, 1999). F. chiloensis is
characterized by an intense and particular aroma (González et al., 2009b), large fruit size
(compared with all other wild species), and remarkable tolerance to the fungus Botrytis
infection (González et al., 2009a). These characteristics make it an important germplasm
source for the development of new cultivars of commercial strawberry (F. x ananassa) with
better organoleptic and nutritional traits. In this regard, some efforts have been done in order
to identify better commercial strawberry genetic populations to recover the genetic resources
with high levels of anthocyanins, phenolic compounds and healthy properties (Aaby et al.,
2012; Diamanti et al., 2012). A chemical study reported six aromatic compounds present in
the fruit, which include ellagic acid, E-cinnamic acid glycosides, an amino acid, and an
anthocyanin (Cheel et al., 2005). Moreover, when chemical composition was compare
between these two strawberry forms and the commercial strawberry (cv. Chandler)
proanthocyanidins, hydrolysable tannins, anthocyanins and flavonol glycosides were found
(Simirgiotis et al., 2009). Quercetin 3-O-glucuronide was the main flavonol glycoside found
in both native strawberry, and both cyanidin malonyl glucoside and pelargonidin malonyl
glucoside were identified at low level. On contrary, highest content of anthocyanin was found
in the commercial strawberry and ellagic acid was the main phenolic found in the native
strawberry (Simirgiotis et al., 2009).
H
EALTH
B
ENEFITS
A
SSOCIATED TO
A
NTHOCYANINS
C
ONSUMPTION
The interest in the study of polyphenols rich diet which includes anthocyanins, has been
intensified after the identification of their potential health benefits (Scalbert and Williamson,
2000). These health-promoting benefits have been deduced, in part, from their antioxidant
properties (Kong et al., 2003). Recent studies have been conducted to elucidate additional
molecular mechanisms involved in polyphenols health benefits. For example, the French
paradox relates the low incidence of coronary heart disease in French people despite a high-
fat diet with the regular consumption of red wine (Renaud and de Lorgeril, 1992). These
additional molecular functions to the antioxidant effects of anthocyanins are produced by
chemical properties different to the antioxidant capacity. Finally, the health’s benefits
promoted by the consumption of fruits and vegetables are not explained by the action of a
single compound or molecule. The phytochemical mixture contained in plants could work
simultaneously to exert health benefits in a collaborative way (Seeram et al., 2004; Zhang et
al., 2008). In the next section we will discuss research advances in the health-benefits effects
of food anthocyanins in diabetes, obesity, cardio-vascular disease and cancer prevention,
neuronal and visual improvement.

Anthocyanins: Food Sources and Benefits to Consumer’s Health 7
Table 4. Summary of anthocyanin effects from fruit extracts on oxidative stress and
glucose metabolism. (Adapted from Sancho and Pastore, 2012)
Anthocyanin source Model Effect
CY-3-glucoside Male Wistar rats/ liver
ischemia-reperfusion
injury
Protective effect against oxidative stress
Plants crude ANT extracts In vitro ↓ α-glucosidase activity
Freeze-dried wild blueberry
powder
Healthy adults/ high-fat
meal
↑ serum antioxidant status
Gamazumi (Viburnum
dilatatum Thunb.) crude
extract
Male Sprague–Dawley
rats/STZ–T2D
↓ oxidative stress markers (TBARS)
↓ plasma glucose
↓ Hb A1c
CY-3-glucoside-rich purple
corn color
Male C57BL/6J
mice/high
fat diet
↓ plasma insulin
↓ plasma glucose
↓ insulin resistance (↑ adipose tissue
TNF-α expression)
CY-3-glucoside, delphinidin-
3-glucoside,
CY-3-galactoside,
pelargonidin-3-galactoside
and CY, delphinidin,
pelargonidin, malvidin,
petunidin
INS-1 832/13 cells ↑ insulin secretion
Vaccinium angustifolium
fruit ethanolic extracts
β TC-tet cells Proliferation of replicating β cells
Cornelian cherry ANT
(delphidin-3-galactoside,
CY-3-galactoside,
pelargonidin 3-galactoside)
Male C57BL/6
mice/high
fat diet
↑ glucose tolerance
↑ glucose-stimulated insulin secretion
ANT-rich black rice extract
Male Sprague–Dawley
rats/high fructose diet
↓ insulin resistance
↓ oxidative stress markers (TBARS and
GSSG)
CY-3-glucoside
Male KK-A
y
mice ↓ plasma glucose
↓ insulin resistance (adipose tissue
upregulated GLUT4 and
downregulated RBP4)
Tart cherries Male Dahl–SS rats ↓ fasting plasma glucose
↓ plasma insulin
↑ antioxidant capacity
Sour cherry juice Diabetic women ↓Hb A1c
Black chokeberry fruit extract
(CY-3-galactoside, CY-3-
arabinoside, CY-3-xyloside,
CY-3-glucoside and others
polyphenols)
Male Wistar rats/low
dose of STZ+diet
fructose and saturated
fat
↓ plasma glucose Jurgoński,
Juśkiewicz, and
Zduńczyk (2008)
↓ plasma oxidative stress markers
(TBARS)
↑ antioxidant enzymes activities (SOD)

Patricio Ramos, Raul Herrera and María Alejandra Moya-León
8
Table 4. (Continued)
Anthocyanin source Model Effect
Pelargonidin Male Wistar rats/STZ–
T2D
↓ insulin resistance
↑ insulin secretion
↓ plasma glucose
↓ plasma oxidative stress markers
(MDA and fructosamine) and
↑antioxidant enzymes activities (SOD
and CAT) ↓ Hb A1c
Strawberry freeze-dried
powder
Women with metabolic
syndrome
↓ serum oxidative stress markers
(MDA, ox-LDL, 4-hydroxynonenal)
no significant differences in fasting
glucose level
Whole blueberry freeze-dried
powder
Male C57BL/6
mice/high
fat diet
↓ insulin resistance (↓ adipose tissue
TNF-α, MCP-1 and IL-6
expression)
↑ adipose tissue antioxidants enzymes
gene expression (GPX)
no effect on modest hyperinsulinemia
Vaccinium arctostaphylos
fruit extract
Male Wistar
rats/aloxan–
T2D
↓ plasma glucose
↑ insulin and GLUT4 expression
↓ α-glucosidase activity
↑ red blood cells antioxidant enzymes
activities
(SOD, CAT and GPX)
Blueberry extract, blueberry
juice, whole blueberry
freeze-dried powder
Male C57BL/6
mice/high
fat diet
↓ fasting plasma glucose
↑ function of β cells
ANT-rich bilberry extract Male KK-A
y
mice ↓ plasma glucose
↓ insulin resistance (activation of
AMPK → ↑ adipose tissue and
skeletal muscle GLUT4 expression, ↓
liver glucose production; ↓
mesenteric adipose tissue RBP4
expression)
Chinese bayberry extract
INS-1 cells and
primary
islets from ICR mice
Inhibition of the H2O2-induced
apoptosis of β cells
Protection of β cells against ROS
Diabetes Control
Type-2 diabetes is a metabolic disorder mainly associated with peripheral insulin
resistance. Insulin, secreted from specialized β-cells of the pancreas, is the molecule in charge
to stimulate the blood glucose transport into cells (Ghosh and Konishi, 2007). Failure in the
insulin’s regulation drives to chronic hyperglycemia, increased oxidative stress and higher
levels of glycosylated proteins including hemoglobin (Hb A1c) (Bonina et al., 2002; Tourrel
et al., 2002). All of these alterations finally lead to the lost of normal function of several
organs.

Anthocyanins: Food Sources and Benefits to Consumer’s Health 9
A large list of experiments have revealed the anti-diabetic properties of the anthocyanins
present in food, which are due to multiple and simultaneous effects of these bioactive
compounds, including: the reduction of blood glucose content, Hb A1c and glucosuria;
preventing free radical production; increasing pancreatic insulin production; and improving
insulin resistance (Guo et al., 2007; Ataie-Jafari et al., 2008; Roy et al., 2008; Grace et al.,
2009; Takikawa et al., 2010). Interestingly, studies in healthy non-diabetic humans
anthocyanin did not showed effects in blood glucose or plasma insulin concentrations, which
suggest that require the pre-existence of a diabetic condition to have an action (Chambers and
Camire, 2003; Kelley et al., 2006; Basu et al., 2009). Properties of anthocyanin-related
glucose metabolism and some associated mechanisms to type 2 diabetes disease are
summarized in Table 4.
A study with cyanidin 3-glucoside (Cy3-g), a purified dietary compound, showed a
reduction in the blood glucose level and improves insulin sensitivity in type 2 diabetes mice
(Sasaki et al., 2007). The results indicate that Cy3-g has anti-diabetic effects due to the
increase of glucose transporter 4 (Glut4) expression and decrease of retinol binding protein 4
(RBP4) in white adipose tissue.
The incorporation of billberry fruit extract (BBE), which contents high levels of
anthocyanins, to the diet decreases the expression of RBP4 in adipose tissue of diabetic mice.
Anthocyanin-rich diet induces the activation of AMP-activated protein kinase (AMPK) in
adipose tissue and skeletal muscle. This activation induces the expression of GLUT4, which
stimulates the uptake of glucose by these tissues through insulin independent mechanisms. In
the liver, the activation of AMPK reduces gluconeogenesis, which is increased in the type 2
diabetes, resulting in a decline of glucose production by the liver (Takikawa et al., 2010).
Additionally, AMPK activation results in significantly reduced liver and serum lipid content
via upregulation of Peroxisome Proliferators-Activated Receptor alpha (PPARα and acyl-
CoA oxidase. All these mechanisms finally increase insulin sensitivity that explains the
anthocyanin-rich diet benefits (Takikawa et al., 2010).
Obesity Prevention
Obesity is defined as an excessive accumulation of adipose tissue caused by the
imbalance of energy intake and expenditure. This condition is associated with various
metabolic disorders representing a strong risk factor for hypertension, heart disease,
hyperlipidemia and type 2 diabetes (Lew and Garfinkel, 1979; Olefsky et al., 1982;
Kopelman, 2000). Studies suggest that consumption of anthocyanins improve the function of
adipocytes and prevent the metabolic syndrome and obesity. In C57BL/6J mice, a Cy3-g -
containing diet prepared from a purple corn extract (2 g/kg) was found to significantly reduce
body fat accumulation induced by high- fat food (60% of energy), when compared with
controls (Tsuda et al., 2003; Tsuda, 2008). An additional study to investigate the
anthocyanin’s effect on gene expression was performed in vitro using isolated adipocytes
(Tsuda et al., 2004). Total RNA isolated was analyzed using GeneChip microarray. After the
treatment of adipocytes with Cy3-glu or cyanidin (Cy), 633 and 427 genes were upregulated,
respectively. Gene expression profile analysis showed the upregulation of hormone-sensitive

Patricio Ramos, Raul Herrera and María Alejandra Moya-León
10
lipase and enhancement of the lipolytic activity. In agreement with this, mice fed with a
semipurified anthocyanin extract from anthocyanin-rich grapes showed improvement in
obesity and triglyceride (TG) metabolism by increasing lipoprotein lipase activity in skeletal
muscle and reducing it in visceral adipose tissue (Lefevre et al., 2008).
Recently, a group of rats showed a decrease in body weight, adipose tissue size and food
intake compared to control when anthocyanins present in black soybean were administered to
rats via intra-gastric (Badshah et al., 2013). The analysis of anthocyanins treated rats brain
proteins showed a significant reduction in the expression of neuropeptide Y (NPY) and
increased c-amino butyric acid receptor (GABA
B1
R) in hypothalamus, which is related to the
reduction of the appetite.
In order to explain the mechanism by which anthocyanins exert their anti-obesity effect,
molecules obtained from grape skin were applied during adipocyte differentiation in 3T3-L1
cells and lipogenesis pathways were studied (Lee et al., 2014). Anthocyanin treated rats
reduced triglyceride accumulation during adipocyte differentiation. This accumulation of TG
was reversed by anthocyanin removal. The mechanism involved was associated to decrease in
gene expression of some transcription factors of lipid metabolism in adipocytes such as liver
X receptor α, sterol regulatory element- binding protein-1c, peroxisome proliferators-
activated receptor-γ, and CCAAT enhancer-binding protein-α. Additionally, the target gene of
these transcription factors such as fatty acid synthase, stearoyl-CoA desaturase-1, and acetyl-
CoA carboxylase α were significantly suppressed by anthocyanins (Lee et al., 2014).
Cardiovascular Disease Prevention
Cardiovascular diseases (CVD) are the 'biggest killers' of the world and now are
considered as the major cause of death among non-transmittable diseases (Butler, 2011). Diet
has been recognized as an important factor of cardiovascular risk. Epidemiological studies
indicate that the consumption of a diet rich in fruits and vegetables is associated with a
decrease in the risk of CVD (Joshipura et al., 2001; Dauchet et al., 2006; He et al., 2007).
The “French paradox” has been proposed as a concept to exemplify the contrast between
low rates of CVD in France, in spite of a high consumption of saturated fats (Renaud et al.,
1992). The frequent consumption of red wine, which contained high levels of polyphenols, is
an important part of the French diet. In addition to the ethanol in wine, there is an important
portion of polyphenols such as flavonoids, and particularly anthocyanins, flavonols and
stilbenes (Flamini et al., 2013).
Epidemiological studies suggest that the daily intake of flavonoids from berries, precisely
anthocyanins, may have cardioprotective properties in humans (Mink et al., 2007; Cassidy et
al., 2011; Cassidy et al., 2013). An association between consumption of strawberry, red wine,
apple, pear among other anthocyanin-rich foods and mortality reduction due to CVD was
found in postmenopausal women (Mink et al., 2007). Another study conducted in
approximately 100,000 women (25−45 years of age) found an association between higher
anthocyanin intake and a 32% decrease in the risk of myocardial infarction after a follow up
period of 18 years (Cassidy et al., 2013). Recently, a study with anthocyanin-rich strawberry
supplementation in humans was performed (Alvarez-Suarez et al., 2014). Healthy human
volunteers were supplemented daily with 500g of fresh strawberries for 1 month. The
strawberry consumption influenced the lipid profile reducing total cholesterol, low-density

Anthocyanins: Food Sources and Benefits to Consumer’s Health 11
lipoprotein cholesterol and TG levels. Additionally, biomarkers of antihemolytic defenses and
platelet function such as spontaneous and oxidative hemolysis and the number of platelet was
significant reduced.
On the other hand, the inhibitory effects on carotid atherosclerosis of a diet with fruits,
berries and vegetables containing anthocyanins were reported in elderly men (Ellingsen et al.,
2008). An inverse association was established between the intima-media thickness of the
carotid artery and dietary intake of vegetables, fruit and berries in elderly men with a high
risk of CVD.
Another report indicates that consumption of anthocyanins inhibits TNFα-induced
inflammation through a reduction of monocyte chemoattractant protein-1 level in human
endothelium (García-Alonso et al., 2009).
Regarding the anthocyanins contained in the red wine, the anthocyanidin malvidin is the
most abundant component in red grape skins and red wines (Watzl et al., 2002). Quintieri et
al. (2013) found that increasing doses of malvidin-rich red grape skin extract induced positive
inotropic and negative lusitropic effects and coronary dilation ( Intracellular malvidin effects
involve phosphatidylinositol 3-kinase (PI3K)/ nitric oxide (NO)/ cyclic guanylate
monophosphate (cGMP)/ protein kinase G (PKG) pathway and this activation could be
associated to Akt, ERK1/2, and GSK3β phosphorylation signal transduction (Quintieri et al.,
2013), in agreement with the cardiac function proposed for these kinases (Clerk et al., 2004).
Anticarcinogenic Activity
Anticarcinogenic properties of anthocyanins have been proposed base on in vitro
evidences and laboratory animal studies. Rats fed with purple corn, which contains higher
anthocyanin levels, decreases the incidence of a multiplicity of colorectal adenomas and
carcinomas induced by 1,2-dimethylhydrazine (DMH) and 2-amino-1-methyl-6-
phenylimidazol (4,5-b) pyridine (PhIP) (Hagiwara et al., 2001). Anthocyanin-rich fraction of
red wines suppresses proliferation of HCT-15 human colon cancer cells and gastric
adenocarcinoma (AGS) cells (Kamei et al., 1998; Shih et al., 2005). Purified cyanidin-3-O-
beta-glucopyranoside induces apoptosis in different human leukemia cell lines (Fimognari et
al., 2004).
Recently, a Chinese group identified and purified two anthocyanins, peonidin-3-
glucoside and cyanidin-3-glucoside, from black rice (Liu et al., 2013). They showed the
ability to inhibit phospho-HER2 and phospho-AKT, confirmed to induce HER2-positive
breast cancer cells apoptosis both in vitro and in vivo. Additionally, both anthocyanin
treatments reduced the tumor size and volume in vivo.
Neuronal Improvement
A study has shown that anthocyanin-rich fruits could have beneficial effects in reversing
the course of neuronal and behavioral aging (Joseph et al., 1999). In addition, the same
authors indicate a beneficial effect of blueberry extracts on the outcome of a
neurodegenerative illness, using transgenic mice as a model for Alzheimer’s disease (AD)

Patricio Ramos, Raul Herrera and María Alejandra Moya-León
12
(Joseph et al., 2003). In the specific case anthocyanins, assays in animal models of AD and
cellular cultures indicate that these molecules can attenuate the neurotoxicity that is induced
by hydrogen peroxide, Aβ peptide and ischemia (Tarozzi, 2007; Tarozzi, 2008; Sul et al.
2009; Tarozzi, 2010; Min et al., 2011).
A protective role was attribute to Cy3-g against Aβ-induced impairment of learning and
memory in rats brain-injected with Aβ peptide through the inhibition of activity for both
GSK-3β and phospho-tau (Qin et al., 2013). Additionally, anthocyanin extracted from
billberries and blackcurrants were assessed in human SH-SY5Y neuroblastoma cells and the
production of reactive oxygen species induced by menadione was significantly decreased
upon treatment. In the same study, the effects of anthocyanin were also investigated in
transgenic AD mice (APdE9) overexpressing mutated human APP (APPswe), a precursor of
Aβ peptide, and presenilin-1 (PS1dE9) proteins, which processes the precursor to produce
Aβ. Mice fed with anthocyanin-rich extracts showed decreased APP processed levels in the
cerebral cortex as compared to APdE9 mice on the control diet. Berry diet alleviated the
spatial memory deficit of aged APdE9 mice as compared to mice with control diet
(Vepsäläinen et al., 2013).
Recently, in cellular models of Parkinson’s disease, treatment with extracts rich in
anthocyanins and proanthocyanidins display a neuroprotective activity against rotenone
neuronal-damage (Strathearn et al. 2014).
Visual Improvement
The consumption of anthocyanins concentrates has a positive effect on night vision, as
healthy volunteers lowered the threshold values to dark adaptation, which indicates an
improvement in eye function under dark condition (Nakaishi et al., 2000). It has been
proposed that the improvement to low-light adaptation performance due to anthocyanins
occurs by promoting the regeneration of rhodopsin (Matsumoto et al., 2003).
On the other hand, studies in animal models suggest that long-term supplementation with
billberry extracts is effective in the prevention of macular degeneration and cataract (Fursova
et al., 2005).
Regarding to the effects on myopia, the administration of anthocyanins for 4 weeks
improved myopia symptoms and contrast sensitivity (Lee et al., 2005). Recently,
anthocyanins from black currant were assessed in two different chick myopia models (Iida et
al., 2013). Cy3-g, cyanidin-3-rutinose and delphinidin-3-glucoside showed positive effects on
the myopia parameters analyzed (Iida et al., 2013)
I
NTAKE
,
B
IOAVAILABILITY AND
M
ETABOLISM OF
A
NTHOCYANINS
The intake of anthocyanins was estimated around 180– 215 mg/day in the United States
people (Kuhnau, 1976). Later, other study calculated the average daily intake based on 24 h
diet and the USDA database for the ‘‘Flavonoid content of Selected Foods’’ (Agricultural
Research Service 2003), and concluded that consumption is much lower and is close to 3
mg/day (Chun et al., 2007). Wu et al. (2006) also estimated the intake for the US population

Anthocyanins: Food Sources and Benefits to Consumer’s Health 13
of 12 mg/day. However, since anthocyanins are habitually restricted to berries, some red
fruits and wine, it should be assumed that not all individuals intake these anthocyanin-rich
products at the same quantity. In Finland, where the consumption of berries is widely
extended, initially the intake was estimated around 47 mg/day in a population study of 2700
Finnish adults (Ovaskainen et al., 2008). However, according to the average consumption of
berries (15 kg/year) in Finland, the daily intake of anthocyanins has been estimated to be 83
mg/day (Heinonen, 2007). A recent study was performed in ten European countries
participating in the European Prospective Investigation into Cancer and Nutrition (EPIC)
study, which considered region, sex, food source and lifestyle, among the other factors
(Zamora-Ros et al., 2011). Results indicated that mean anthocyanidin intake ranged from
18.73 mg/day (Granada, Spain) to 64.88 mg/day (Turin, Italy). A detailed report of
anthocyanin composition of each group diet and class of fruit and vegetables consumption are
also summarized in the work of Zamora-Ros et al. (2011).
Despite information related to health-promoting properties of anthocyanins, the
evaluation of their real bioactivity is necessary to understand their metabolism after
consumption in human. Bioavailability studies have also demonstrated that anthocyanins are
absorbed in stomach (Passamonti et al., 2003; Talavera et al., 2003) and in the small intestine
(Miyazawa et al., 1999), being detected in plasma and urine as their parental form or
methylated, glucuronidated or sulphated compounds (Kay et al., 2005).
Anthocyanins after absorption, reach a maximum concentration in plasma (Cmax)
oscillating from 1.4 to 200 nM for doses of 10 and 720 mg of anthocyanins, respectively. The
Cmax is reached after 45 min to 4 h after ingestion of a meal with anthocyanins, depending
on the conditions of the trial. When the ingested considered anthocyanin alone and after
overnight fasting, the Cmax is reached after only 1 h (Matsumoto et al., 2001). However,
when anthocyanins are ingested together with other food, 1.5 h is required for absorbing
(Nielsen et al., 2003), and 4 h in the case of anthocyanins ingest in meals with a very high fat
content (Mazza et al., 2002).
A second via of anthocyanin metabolism has also been described. In vitro studies
indicated that phenolic acids, such as protocatechuic, syringic, vanillic, and p-hydroxybenzoic
acids, are the main metabolites of anthocyanins degradation by gut microflora (Aura et al,.
2005; Keppler and Humpf, 2005; Fleschhut et al., 2006; Forester and Waterhouse, 2008).
Recently, measures of anthocyanin metabolites reveal that are abundant in human urine after
5 days post-consumption (Kalt et al., 2014). After intake of blueberry juice, the anthocyanins
parent represent 4%, while anthocyanin metabolites were 96% of total urinary anthocyanin
after 24 h.
C
ONCLUSION
In conclusion, several evidences which strongly suggest a beneficial role of anthocyanins
on health were reported in this review. These molecules coming from plants are ingested by
the consumption of several fruits, being mostly abundant in berries. The potential benefits and
health-promoting effects are broad, ranging from antioxidant activity to neuronal function
improving among the many others reviewed. The beneficial effects have been observed and
verified in clinical trials. All these evidences indicate that regular consumption of colored

Patricio Ramos, Raul Herrera and María Alejandra Moya-León
14
edible vegetables, and specifically berry fruits, promotes improvement in health and quality
of life of human beings.
A
CKNOWLEDGEMENT
This work is in the frame of project Anillo ACT 1110.
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