FOOD & NUTRITION RESEARCH, 2017

VOL. 61, 1361779

https://doi.org/10.1080/16546628.2017.1361779

REVIEW ARTICLE

Anthocyanidins and anthocyanins: colored pigments as food, pharmaceutical

ingredients, and the potential health benefits
a,b a,b a c
Hock Eng Khoo , Azrina Azlan , Sou Teng Tang and See Meng Lim
a
Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Selangor Darul Ehsan, Malaysia;
b
Research Centre of Excellence for Nutrition and Non-communicable Diseases, Faculty of Medicine and Health Sciences, Universiti Putra
Malaysia, Selangor Darul Ehsan, Malaysia; cNutritional Sciences Program, School of Healthcare Science, Faculty of Health Sciences, Universiti
Kebangsaan Malaysia, Kuala Lumpur, Malaysia

ABSTRACT ARTICLE HISTORY

Anthocyanins are colored water-soluble pigments belonging to the phenolic group. The pig- Received 10 April 2017
ments are in glycosylated forms. Anthocyanins responsible for the colors, red, purple, and blue, Accepted 23 July 2017
are in fruits and vegetables. Berries, currants, grapes, and some tropical fruits have high antho- KEYWORDS
cyanins content. Red to purplish blue-colored leafy vegetables, grains, roots, and tubers are the Anthocyanin; colorant;
edible vegetables that contain a high level of anthocyanins. Among the anthocyanin pigments, disease; health benefit;
cyanidin-3-glucoside is the major anthocyanin found in most of the plants. The colored antho- pigment
cyanin pigments have been traditionally used as a natural food colorant. The color and stability of
these pigments are influenced by pH, light, temperature, and structure. In acidic condition,
anthocyanins appear as red but turn blue when the pH increases. Chromatography has been
largely applied in extraction, separation, and quantification of anthocyanins. Besides the use of
anthocyanidins and anthocyanins as natural dyes, these colored pigments are potential pharma-
ceutical ingredients that give various beneficial health effects. Scientific studies, such as cell
culture studies, animal models, and human clinical trials, show that anthocyanidins and antho-
cyanins possess antioxidative and antimicrobial activities, improve visual and neurological health,
and protect against various non-communicable diseases. These studies confer the health effects
of anthocyanidins and anthocyanins, which are due to their potent antioxidant properties.
Different mechanisms and pathways are involved in the protective effects, including free-radical
scavenging pathway, cyclooxygenase pathway, mitogen-activated protein kinase pathway, and
inflammatory cytokines signaling. Therefore, this review focuses on the role of anthocyanidins
and anthocyanins as natural food colorants and their nutraceutical properties for health.
Abbreviations: CVD: Cardiovascular disease VEGF: Vascular endothelial growth factor

Introduction flowers are red hibiscus, red rose, red pineapple sage,
red clover, and pink blossom. These red flowers are
Anthocyanins are blue, red, or purple pigments found in
edible. Blue (cornflower, blue chicory, and blue rosem-
plants, especially flowers, fruits, and tubers. In acidic
ary) and purple (purple mint, purple passion flower,
condition, anthocyanin appears as red pigment while
purple sage, common violet, and lavender) flowers are
blue pigment anthocyanin exists in alkaline conditions.
the common edible flowers. Some of these flowers have
Anthocyanin is considered as one of the flavonoids
been traditionally used as folk medicine, as colorants,
although it has a positive charge at the oxygen atom of
and as food. In addition to traditional usage, red, purple,
the C-ring of basic flavonoid structure. It is also called
and blue-colored fruits are commonly consumed for
the flavylium (2-phenylchromenylium) ion. The general
their beneficial effects. The colored pigments of antho-
molecular structure of anthocyanin is shown in Figure 1.
cyanin from berries, blackcurrants, and other types of
The stability of anthocyanin is dependent on pH, light,
red to blue-colored fruits are strong antioxidants.
temperature, and its structure [1].
Moreover, anthocyanin-rich black carrot, red cabbage,
Anthocyanins are commonly found in flowers and
and purple potato are potential functional foods that
the fruits of many plants. Most of the red, purple, and
have been consumed for prevention of diseases.
blue-colored flowers contained anthocyanins. Red

CONTACT Azrina Azlan azrinaaz@upm.edu.my Department of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, Universiti Putra
Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia.
All authors contributed equally to this review.
© 2017 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted
use, distribution, and reproduction in any medium, provided the original work is properly cited.
2 H. E. KHOO ET AL.

Figure 2. Two-dimensional structure of flavylium ion.

Figure 1. Basic anthocyanin structure.

anthocyanidins. The conjugated bonds of anthocyanins
result in red, blue, and purple-colored plants.
Cyanidin, delphinidin, pelargonidin, peonidin, mal-
Anthocyanins found in plants have a wide range of
vidin, and petunidin are the most common anthocya-
usage. Blue, red, and purple colored pigments extracted
nidins distributed in the plants. The distribution of
from flowers, fruits, and vegetables are traditionally
these anthocyanidins in fruits and vegetables is 50%,
used as dye and food colorant. Besides being used as
12%, 12%, 12%, 7%, and 7%, respectively [3]. Their
natural colorants, some of the anthocyanin-rich flowers
molecular structures are shown in Figure 3. In nature,
and fruits have been traditionally used as medicine to
cyanidin is a reddish-purple (magenta) pigment. It is
treat various diseases. On the other hand, plant antho-
the major pigment in berries [4] and other red-colored
cyanins have been widely studied for their medicinal
vegetables such as red sweet potato and purple corn
values. Anthocyanins possess antidiabetic, anticancer,
[5]. Delphinidin has a chemical characteristic similar to
anti-inflammatory, antimicrobial, and anti-obesity
most of the anthocyanidins. It appears as a blue-red-
effects, as well as prevention of cardiovascular diseases
dish or purple pigment in the plant. The blue hue of
(CVDs) [2]. Therefore, anthocyanins extracted from
flowers is due to the delphinidin pigment [6].
edible plants are potential pharmaceutical ingredients.
Pelargonidin differs from most of the anthocyanidins.
In nature, it appears as red-colored pigment [7].
Types of anthocyanin in plants Pelargonidin gives an orange hue to flowers [8] and
red to some of the fruits and berries [9].
Anthocyanin is one of the subclasses of phenolic phy-
Methylated anthocyanidin such as peonidin is
tochemicals. Anthocyanin is in the form of glycoside
another type of anthocyanidin abundantly found in
while anthocyanidin is known as the aglycone.
plants. It has the visible color magenta [7]. Peonidin
Anthocyanidins are grouped into 3-hydroxyanthocya-
is abundantly found in berries, grapes, and red wines.
nidins, 3-deoxyanthocyanidins, and O-methylated
Malvidin is another O-methylated anthocyanidin. It
anthocyanidins, while anthocyanins are in the forms
has a purple visible color, and is abundant in blue-
of anthocyanidin glycosides and acylated anthocyanins.
colored flowers, especially Summer Wave Blue [10].
The most common types of anthocyanidins are cyani-
Malvidin is also the major red pigment in red wine
din, delphinidin, pelargonidin, peonidin, petunidin,
[11]. It appears as darker dusty red in matured red
and malvidin. Acylated anthocyanins are also detected
wines [12]. Petunidin is a methylated anthocyanidin.
in plants besides the typical anthocyanins. Acylated
It is a dark red or purple pigment that is soluble in
anthocyanin is further divided into acrylated anthocya-
water [7]. Petunidin has been detected in blackcurrants
nin, coumaroylated anthocyanin, caffeoylated antho-
[13] and purple petals of flower [14].
cyanin, and malonylated anthocyanin.
Anthocyanin is derived from flavonol, and it has the
basic structure of flavylium ion, that is a lack of a ketone Color and stability of anthocyanin pigments
oxygen at the 4-position (Figure 2). The empirical for-
Stability of anthocyanin color based on pH
mula for flavylium ion of anthocyanin is C15H11O+ with
a molecular weight of 207.24724 g/mol. On the other Stability of anthocyanins is dependent on the type of
hand, anthocyanins are the glycosylated form of anthocyanin pigment, copigments, light, temperature,
 FOOD & NUTRITION RESEARCH 3

Cyanidin Delphinidin

Pelargonidin Peonidin

Malvidin Petunidin

Figure 3. Major anthocyanidins in plants.

pH, metal ions, enzymes, oxygen, and antioxidants This is due to the kinetic and thermodynamic competi-
[15]. Anthocyanidinʼs stability is also influenced by tion between the hydration reaction of flavylium ion
the B-ring in the anthocyanidin structure and the pre- [18]. This blue quinonoidal species is unstable at lower
sence of hydroxyl or methoxyl groups [3]. These pH. At pH 4–5, an anthocyanin solution has very little
groups are known to decrease anthocyanidin stability hue due to the small amount of flavylium cation and
in a solution. In this section, we discuss the stability of quinonoidal anion [19]. At neutral pH, resonance-sta-
anthocyanin and its color changes in different pH bilized quinonoid anions (color purple of anthocya-
conditions. nins) are formed from further deprotonation of the
The color of anthocyanins is depending on the pH quinonoidal species.
of the solution. This is because of the molecular struc- The predicted pKa of a flavylium cation in a buffered
ture of anthocyanins having an ionic nature [15]. In solution is 1–3, pKa of quinonoidal base and chalcone
acidic condition, some of the anthocyanins appear red. is 4–5, and the pKa of quinonoidal and chalcone mono-
Anthocyanins have a purple hue in neutral pH while anions is 7.5–8.0 [20]. The flavylium cation and qui-
the color changes to blue in an increasing pH condi- nonoidal base appear red, while quinonoidal
tion. The red-colored pigments of anthocyanins are monoanion and dianion can be seen as purple and
predominantly in the form of flavylium cations [7]. blue in aqueous solution. At a lower pH solution (pH
These anthocyanins are more stable at a lower pH <3), cyanidin appears to be red, violet at pH 7–8; it is
solution. At lower pH, the flavylium cation formed blue in color at a very high pH (pH >11) [21]. Peonidin
enables the anthocyanin to be highly soluble in water. (3-O-methylated anthocyanin) has cherry red hue at
The decrease in water concentration increases the rate low pH, but its color changes to deep blue at pH 8. It is
of deprotonation of the flavylium cation, thus reducing different from most of the anthocyanidins because it
color stability [16]. Apart from the pH, anthocyanin- has higher stability at high pH than cyanidin, delphi-
tannin polymerization could also increase the color nidin, and pelargonidin.
stability at a lower pH [17]. Peonidin is also stable at high pH, therefore, the
At increasing pH conditions, colorless carbinol blue hue of flowers is from peonidin [22]. On the
pseudobase and chalcone structures are formed, fol- contrary, peonidin is found in red-purplish fruits at
lowed by formation of anionic quinonoidal species. lower pH conditions [23]. Petanin (petunidin-3-[6-O-
4 H. E. KHOO ET AL.

(4-O-E-p-coumaroyl-O-α-l-rhamnopyranosyl)-β-d- as blanching, in the food processing industry can pre-

glucopyranoside]-5-O-β-d-glucopyranoside), is also vent oxidation of anthocyanins by polyphenol oxidase.
one of the acylated anthocyanins which are stable at
higher pH [18]. Acylation of anthocyanin increases the
Anthocyanin pigments as food colorants and
proportion of flavylium cations and helps to maintain
additives
the redness of anthocyanin pigment even at increasing
pH [19]. The use of natural colorant and additives in processed
Most of the anthocyanin pigments have a high sta- foods and beverages is important for increasing con-
bility in acidic conditions compared with bases, and sumer acceptability of these products. Anthocyanins
degradation occurs at higher pH. Cyanidin and delphi- are some of the natural colored pigments extracted
nidin are the examples of anthocyanidin which is stable from plants, which have an attractive hue.
in acidic condition. However, there has some excep- Anthocyanins extracted from plants are red, blue, and
tion. Although delphinidin is a purple-colored pig- purple pigments. These pigments are the natural color-
ment, the blue hue of flower could be due to the ants with low to no toxicity. Natural colorants are
delphinidin pigment in higher pH conditions [6]. At somehow safe to be consumed even at higher doses
higher pH or alkaline conditions, for example, petanin compared to synthetic colorants. Anthocyanins, as nat-
is resistant to degradation even at pH 8 [18]. Due to the ural colorants, have value-added properties [29]. These
high stability of petanin at high pH, it is suitable for use properties are antioxidants, as nutraceutical and many
as a colorant for herbal beverages. health benefits, such as an antimicrobial effect and
prevention of chronic diseases.
Effect of copigmentation and temperature on color
change Solubility of anthocyanin pigments
Copigmentation and temperature are known to influ- Anthocyanins occur as flavylium ions in grapes and
ence the color change of anthocyanins in a solution in wines. During digestion, the flavylium ion is converting
addition to different pH conditions. Copigmentation of to carbinol pseudo-base, quinoidal-base, or the chal-
anthocyanin aglycone is referred to as a phenomenon cone at increasing pH before being absorbed into the
where anthocyanidins are reinforced by metallic ions or blood system [30]. As mentioned earlier, anthocyanin
flavonoids. Copigmentation helps to stabilize the color pigment appears to be red in acidic conditions and blue
of the leaves, flowers, and fruits of the plant [24]. The to purple in alkaline solutions. These colored pigments
addition of a metal ion helps stabilization of carbinol are commonly extracted from flowers, berries, black-
pseudo-base structures in the equilibrium mixture of currant, and purple-colored fruits and vegetables. Also,
delphinidin at appropriate pH values and provides a water is the typical extraction medium for isolation of
blue hue [25]. Also, color changes of flower anthocya- anthocyanin pigments. Moreover, some food proces-
nins are due to the copigmentation of anthocyanidins sing factories use alcoholic solutions to extract antho-
with flavonoids, which increases the color intensity of cyanin pigments. It is because anthocyanins are soluble
the flower [3]. In addition, glycosylation and acylation in both water and most of the organic solvents. On the
increase the color strength of anthocyanin. contrary, anthocyanin is not soluble in the apolar
Anthocyanins are less stable at higher solution tem- organic solvent. It is also not stable in alkaline or
peratures. A previous study reports that heat treatment neutral solutions.
at a maximum of 35°C reduced the total anthocyanin At a lower pH or in an acidic condition, anthocya-
content in the common grape to less than half the nidins such as cyanidin is highly soluble in water due
amount in control berries at 25°C [26]. At up to 40° to the formation of flavylium cation which appears as
C, the color of anthocyanin changes from red to orange red [19]. Acidic conditions maintain the stability of
although the pH of the solution was low [27]. In con- flavylium ion and increase the intensity of the red
trast, heat treatment of an anthocyanin-rich extract hue of the anthocyanin pigment. This characteristic
solution may not cause a degradation of anthocyanin makes it a good candidate as a colorant, where the
pigments. This is because the extract commonly con- red anthocyanin pigment is highly stable in acidic
tains phenolic compounds that are enzymatically aqueous solutions. It is best used as a colorant for
degraded by polyphenol oxidase. In addition, mild red-colored beverages. Deprotonation of the flavylium
heat treatment of the extract to up to 50°C has been ion occurs at increasing pH, where the quinoidal bases
shown to inactivate the enzymatic reaction [28]. favor alcoholic solution [16]. For example, the colors of
Therefore, mild heat treatment of raw materials, such red wine can be visualized as purplish red at lower pH
 FOOD & NUTRITION RESEARCH 5

or bluish-violet at higher pH. The typical range of pH Extraction and identification of anthocyanins
in red wine is 2.5–4.0 [17]. In an alcoholic solution, the
The use of organic solvents such as methanol and
red flavylium cation substantially converts to blue qui-
ethanol to extract anthocyanin pigments causes a toxi-
nonoidal species which makes the red wine looks blu-
city issue. Although ethanol is considered as a generally
ish-violet.
safe extraction medium, isolation of anthocyanins
Anthocyanin aglycone has higher solubility in alco-
using water-based extraction is consider a greener
hol than its glucoside, whereas glycosylated anthocya-
way. Subcritical water-based extraction is one of the
nin is highly soluble in water [31]. The polyphenolic
methods that have been tested for the extraction of
structure of anthocyanin adds a hydrophobic charac-
anthocyanins from berries. This extraction technique
teristic to it, and makes it soluble in organic solvents,
uses acidified water (0.01% HCl, pH ~2.3) that is sub-
such as ethanol and methanol. Besides the flavylium
jected to high temperatures between 110–160°C under
cation, the solubility of anthocyanidin in water could
a constant pressure of 40 bars [38]. It is a highly
be due to the 3-hydroxyl group in the C-ring of antho-
efficient technique for extraction of anthocyanins
cyanidin that always linked to sugar(s), which forms a
from fruit. Anthocyanin pigments also can be extracted
stable anthocyanin in water. Among the anthocyani-
by the addition of sulfur dioxide to water for stabilizing
dins, delphinidin is the most soluble in methanol,
the anthocyanin structure with an enhanced diffusion
followed by water, ethanol, and acetone [32]. As com-
coefficient of anthocyanin molecules through the solid
pared to delphinidin, cyanidin has lower solubility in
[39]. This increases the solubility of anthocyanins from
methanol. It is because a low yield of cyanidin has been
the plant during extraction with water.
extracted from grape skins using methanol [33]. Also,
Anthocyanins are extracted from plants as a crude
delphinidin has higher solubility in water compared to
mixture. For that reason, separation or isolation of
malvidin because malvidin has lower polarity than
specific type of anthocyanin is needed for a specific
delphinidin [34].
purpose. Separation and identification of anthocya-
Malvidin is highly soluble in water compared with
nins can be done by various chromatographic meth-
methanol and ethanol due to the static dipole moment
ods. These include thin layer chromatography, high
of malvidin in water being higher than in methanol
speed countercurrent chromatography, high-perfor-
and ethanol [35]. Its solubility in water reduces with
mance liquid chromatography, cellulose column
increasing degrees of acylation [12]. At a high pH
chromatography, and reversed-phase ion-pair chro-
condition (pH >7), syringic acid is released from the
matography, as well as gas chromatography. In the
breakdown of acylated malvidin. The diffusion coeffi-
early days, cellulose column chromatography is used
cients of both malvidin-3-glucoside and peonidin-3-
for the separation of anthocyanin mixtures, but diffi-
glucoside in water at room temperature (25°C) are
culties may be encountered in applying this techni-
similar, even at increasing temperature [36]. Although
que in the presence of large amounts of other
no study compares the solubility of these six anthocya-
flavonoid materials. Purification of anthocyanin car-
nins in water, a decreasing polarity of anthocyanidins
ried out by countercurrent chromatography is also
has been reported in the order of delphinidin, cyanidin,
too expensive to be popular [40].
petunidin, pelargonidin, peonidin, and malvidin [37].
Lately, macroporous adsorption resin is used in the
In actual fact, malvidin, peonidin, and petunidin are
purification of phenolic pigments because AB-8 resin is
less soluble in water compared to cyanidin, delphini-
a kind of macroporous resin specially invented for
din, and pelargonidin. It could be due to these antho-
purification of flavonoid [41]. In addition to separation
cyanidins have one or more hydrophobic methoxy
and identification of anthocyanins, quantification of
groups at positions 3ʹ, 5ʹ, and 3ʹ and 5ʹ of the B-ring.
these compounds is commonly done by various chro-
The solubility of anthocyanins in water increases at
matographic methods. High-performance liquid chro-
lower pH values where strong protonation occurs [16].
matography is the most used method in quantification
The addition of HCl to alcohol increases the solubility
of anthocyanins. However, gas chromatography has
of anthocyanins [35]. Malonylation of anthocyanidin
been applied in the quantification of anthocyanins
also enhances its solubility in water and stabilizes its
[42]. Although gas chromatography is invented speci-
structure [31]. Therefore, malonylation of anthocyanin
fically for identification and quantification of hydro-
aglycone preserves its pigment color for the use as food
carbon, the use of mass spectrometry enables the
colorant. Nevertheless, the color of anthocyanin is
determination of anthocyanins using gas
greatly dependent on the number of hydroxyl groups
chromatography.
attached to the B-ring.
6 H. E. KHOO ET AL.

Anthocyanins and anthocyanidin in plants The use of anthocyanin-based colorants in yogurt

drink and some mixed fruit juice is becoming more
Anthocyanins are found abundant in plants, including
popular. Some companies did use synthetic dyes in
red-purplish or red to blue-colored fruits, leaves, flow-
their products. However, these synthetic dyes may be
ers, roots, and grains. Types of anthocyanin and antho-
toxic if overconsumed. Recently, acylated anthocyanins
cyanidin have been determined in fruits and vegetables.
are food colorants used in the food industry due to
As shown in Table 1, cyanidin-3-glucoside is the most
their high stability over nonacylated anthocyanins [64].
abundant anthocyanin determined in fruits and vege-
A high level of nonacylated anthocyanins are produced
tables. In plants, cyanidin-3-glucoside is formed as the
from certain fruits, such as elderberry and barberry, at
consequence of low pH [54]. All berries that contain
relatively low cost. These commodities have potential
glycosides of cyanidin probably do so due to the acidic
as colorants for use in the food industry.
nature of the berries. Malvidin, peonidin, and petuni-
din are not commonly found in berries. These antho-
cyanidins are in methylated forms, therefore, the Nutraceutical and pharmaceutical effects of
pigments are not commonly found in red berries. A anthocyanins
possible reason is that methylated anthocyanidin has
Anthocyanin is one of the bioactive components as
lesser reddening effect than the non-methylated struc-
nutraceutical and traditional medicine. It has been
ture. Moreover, these anthocyanidins are typically
traditionally used as a phytopharmaceutical, appetite
detected in blue-colored fruit.
stimulant, choleretic agent, and for treatment of many
Petunidin is the anthocyanidin formed in most
other diseases. These colored pigments are potent
fruits. Other than the fruits and tomato as fruit vege-
nutraceutical or pharmaceutical ingredients. As a
table, as well as flowers, petunidin is not commonly
nutraceutical, the bioavailability of anthocyanin is the
determined in purple-colored leaves, roots, and grains.
key factor for maintaining good health and for preven-
Although most of these purple-colored vegetables con-
tion of diseases.
tain petunidin and its glycosides, these pigments are
The low bioavailability of anthocyanins causes low
not well-known for the potential health benefits.
absorption of these compounds into the blood circulat-
Similar to petunidin, malvidin is also one of the less
ing system and a high excretion rate of anthocyanins in
popular anthocyanidins. Nevertheless, malvidin is a
urine and feces, thus reducing the efficacy of anthocya-
potent antioxidant with high bioavailability [62].
nins in scavenging free radical. Anthocyanin with a
high bioavailability efficiently reduces cellular lipid
peroxidation, hence reducing the risk of many diseases.
Potential uses of anthocyanin pigments
Until now, limited reports on the bioavailability of
Anthocyanins extracted from plants have been used as major anthocyanins are available for comparison. In
food additives. Food additive, E163, is one of the com- this review, bioavailability of selected anthocyanidin
mercial additives derived from fruit anthocyanin such and anthocyanin will be discussed.
as grape skin. It is a purple food additive for use in Among the major anthocyanins, the bioavailability
producing purple-colored jam, confectionaries, and of cyanidin-3-glucoside and malvidin-3-glucoside is
beverages. Recently, synthetic food dyes attracted pub- the most reported. The relative bioavailability of red
lic concern regarding safety and the adverse effect on wine anthocyanins has been reported previously, where
human health, particularly neurological functions and the glycosides of peonidin had the highest relative
behavioral effects. A clinical trial involving a total of bioavailability, followed by the glycosides of cyanidin,
153 three-year-old and 144 eight to nine-year-old chil- malvidin, delphinidin, and petunidin [65]. Although
dren suggested that artificial colorants that contained a most red wine anthocyanins have low bioavailability,
mixture of sunset yellow (E110), carmoisine (E122), increased consumption of these anthocyanins may help
tartrazine (E102), ponceau 4R (E124), quinoline yellow to increase the efficacy. However, the side effects of
(E104), and Allura red AC (E129) when combined in overconsumption of anthocyanin remain unknown.
the diet with sodium benzoate (E211), resulted in a A study has determined the absorption of malvidin-
significant increase of hyperactivity in normal children 3-glucoside after ingestion of red wine and red grape
and aggravated the condition as well at least up to juice [62], where malvidin-3-glucoside was found in
middle childhood [63]. This finding has drawn great plasma and urine within 3 and 6 h of consumption of
interest in exploring natural food colorants such as these beverages. The result also demonstrates that the
anthocyanin as a promising alternative to synthetic bioavailability of malvidin-3-glucoside was about two
food dyes. times higher after consumption of red grape juice
 FOOD & NUTRITION RESEARCH 7

Table 1. Anthocyanins and anthocyanidins in fruit, vegetables, and grains.

Types of anthocyanin and anthocyanidin in fruit
Acai berry (Euterpe oleracea Martius) – whole fruit [43]
cya-3-glu, cyan-3-rut, del-3-gal, del-3-glu, del-3-rut, peo-3-glu
Berry (Berberis lycium Royle) – whole fruit [44]
cya-3,5-dihex, cya-3-gal, cya-3-glu, cya-3-lat, cya-3-rut, del-3-glu, mal-3,5-dihex, pel-3,5-diglu, pel-3-pentoxilhex, pel-3-rut, pel-hex, peo-3-rut
Bilberry (Vaccinium myrtillus L.) – whole fruit [45]
cy-3-ara, cya-3-gal, cya-3-glu, del-3-ara, del-3-glu, del-3-gal, mal-3-ara, mal-3-gal, mal-3-glu, peo-3-ara, peo-3-gal, peo-3-glu, pet-3-ara, pet-3-gal, pet-3-glu
Blackberry (Rubus fruticosus L.) – whole fruit [46,47]
cya-3-glu, cya-3-rutl del, mal, pel, pel-3-glu, peo
Blackcurrant (Ribes nigrum L.) – whole fruit [46]
cya-3-glu, cya-3-rut, del-3-glu, del-3-rut
Blueberry (V. corymbosum L.) – whole fruit [46]
cya-3-ara, cya-3-gal, cya-3-glu, del-3-ara, del-3-gal, del-3-glu, mal-3-ara, mal-3-gal, mal-3-glu, peo-3-gal, peo-3-glu, pet-3-ara, pet-3-gal, pet-3-glu
Cranberry (V. oxycoccos L.) – whole fruit [46]
cya-3-ara, cya-3-gal, peo-3-ara, peo-3-gal
Dabai (Canarium odonthophyllum Miq.) – skin [48]
cya-3-glu, cya-3-gal, cya-3-ara, cya-3-sop, cya-3-rut, del-3-glu, del-3-gal, mal-3,5-diglu
Maqui berry [Aristotelia chilensis (Mol.) Stuntz] – whole fruit [49]
cya-3-glu, cya-3-sam, cya-diglu, cya-sam-glu, del-3-glu, del-3,5-diglu, del-3-sam, del-3-sam-5-glu
Nitratia (Nitraria tangutorun Bobr.) – seed [50]
cya-3-O-(caffeoyl)-diglu, cya-3-O-(cis-p-coumaroyl)-diglu, cya-3-O-(trans-p-coumaroyl)-diglu, cya-3-diglu, del-3-O-(p-coumaroyl)-hexose, del-3-O-(caffeoyl)-
diglu, pel-3-O-(p-coumaroyl)-diglu, pel-3-O-diglu
Oregon grape (Mahonia aquifolium (Pursh) Nutt) – whole fruit [51]
cya-3-glu, cya-3-rut, del-3-glu, del-3-rut, mal-3-glu, pel-3-glu, peo-3-glu
Pomegranate (Punica granatum cv. Mollar de Elche) – edible flesh [52]
cya-3,5-diglu, cya-3-glu, cya-pen, del-3,5-di-glu, del-3-glu, pel-3,5-di-glu, pel-3-glu
Raspberry (Rubus idaeus L.) – whole fruit [46]
cya-3-glu, cya-3-rut, cya-3-sop
Red grape (Vitis vinifera L.) from different cultivars – whole fruit [53]
cya-3-O-glu, del-3-O-glu, mal-3-O-acetylglu, mal-3-O-glu, mal-3-p-coumarylglu, peo-3-O-acetylglu, peo-3-O-glu, peo-3-p-coumarylglu, pet-3-O-glu
Types of anthocyanin and anthocyanidin in vegetables and grains
Black carrots (Daucus carota ssp. sativus var. atrorubens Alef.) [55]
cya-3-xylosyl-glucosyl-gal, cya-3-xylosyl-gal, cya-3-xylosyl-glucosyl-gal-coumaric acid, cya-3-xylosyl-glucosyl-gal-ferulic acid, cya-3-xylosyl-glucosyl-gal-
sinapic acid
Black soybean [Glycine max (L.) Merrill] [56]
cya-3-gal, cya-3-glu, del-3-glu, peo-3-glu.
Purple corn (Zea mays L.) [57]
cya-3-(6″-malonylglu), cya-3,5-diglu, cya-3-dimalonylglu, cya-3-glu, cya-3-malonylglu, cya-3-malonylglu, cya-3-succinylglu, pel-3-(6″-maolonylglu), pel-3-
dimalonylglu, pel-3-glu, peo-3-(6″-malonylglu), peo-3-dimalonylglu, peo-3-glu
Purple sweet potato (Ipomoea batatas L.) [58]
cya-3-(caffeoyl sop)-5-glu, cya-3-(caffeoyl-feruloyl sop)-5-glu, cya-3-(caffeoyl-p-hydroxybenzoylsop)-5-glu, cya-3-(dicaffeoylsop)-5-glu, cya-3-(feruloyl sop)-
5-glu, cya-3-(p-hydroxybenzoylsop)-5-glu, cya-3-sop-5-glu, pel-3-(caffeoyl-feruloyl sop)-5-glu, peo-3-(caffeoyl sop)-5-glu, peo-3-(caffeoyl-feruloyl sop)-5-
glu, peo-3-(caffeoyl-p-coumaroyl sop)-5-glu, peo-3-(caffeoyl-p-hydroxybenzoylsop)-5-glu, peo-3-(dicaffeoylsop)-5-glu, peo-3-(feruloyl sop)-5-glu, peo-3-
(feruloyl-p-coumaroyl sop)-5-glu, peo-3-(feruloyl-p-hydroxybenzoylsop)-5-glu, peo-3-(p-hydroxybenzoylsop)-5-glu, peo-3-sop-5-glu
Red cabbage (Brassica oleracea L. var. capitata L.) [59]
cya-3-glu, cya-3-rut, del-3-glu, del-3-rut, cya-3-diglu-5-glu, cya-3-(caffeoyl)(p-coumaroyl) diglu-5-glu, cya-3-(sinapoyl) diglu-5-glu, cya-3-(caffeoyl-sinapoyl)
diglu-5-glu, cya-3-(p-coumaroyl)(sinapoyl) triglu-5-glu, cya-3-(feruloyl)(sinapoyl) triglu-5-glu, cya-3-(p-coumaroyl) diglu-5-glu, cya-3-(sinapoyl) diglu-5-
glu, cya-3-(p-coumaroyl)(sinapoyl) diglu-5-glu, cya-3-(feruloyl)(sinapoyl) diglu-5-glu, cya-3-(sinapoyl)(sinapoyl) diglu-5-glu, cya-3-(sinapoyl) diglu-5-
(sinapoyl) glu
Rice (Oryza sativa L. cv. Heugjinju) [60]
cya, cya-3-glu, peo-3-glu
Transgenic purple tomato (Solanum lycopersicum L. cv. Del/Ros1) [61]
del-3-(caffeoyl)-rut-5-glu, del-3-(feruloyl)-rut-5-glu, del-3-(trans-coumaroyl)-rut-5-glu, mal-3-(feruloyl)-rut-5-glu, mal-3-(p-coumaroyl)-rut-5-glu, pet-3-
(feruloyl)-rut-5-glu, pet-3-(trans-coumaroyl)-rut-5-glu
*ara: arabinoside, cya: cyanidin, del: delphinidin, mal: malvidin, pel: pelargonidin, pen: pentoside, peo: peonidin, pet: petunidin, gal: galatoside, glu:
glucoside, hex: hexoside, lat: lathyroside, rut: rutinoside, sam: sambubioside, sop: sophoroside

compared to red wines. Another study also found that In fact, anthocyanins are more bioavailable than
the urine anthocyanin level in the urine of six healthy previously assumed. Literature shows that the relative
volunteers who drank 300 ml of red wine (218 mg bioavailability of cyanidin-3-glucoside was
anthocyanins) reached a peak within 6 h of wine con- 12.38 ± 1.38%, where 5.37 ± 0.67% was excreted in
sumption [66]. Another study reports on drinking of urine and 6.91 ± 1.59% in the breath within 48 h after
11 g of elderberry concentrate containing 17.2% w/w of oral ingestion [68]. The reported metabolites of cyani-
anthocyanins (mainly containing cyanidin-3-glucoside din-3-glucoside after digestion by the human body are
and cyanidin-3-sambubioside) on an empty stomach phenolic acid, and phenolic conjugates, hippuric, phe-
[67]. Based on the results obtained, low bioavailability nylacetic, and phenylpropenoic acids. On the other
of anthocyanins was suggested by the researchers. hand, delphinidin-3-rutinoside, cyanidin-3-rutinoside,
8 H. E. KHOO ET AL.

delphinidin-3-glucoside, and cyanidin-3-glucoside hydroxylation and methoxylation substantially

from blackcurrant have been found directly absorbed increase the antioxidant activity [106].
into the human blood circulating system and their In fact, anthocyanidin has a higher ORAC value
glycosylated forms are excreted into urine [69]. This than anthocyanin. One of the possible reasons is antho-
observation is also supported by a previous study that cyanin aglycone is very unstable and highly reactive
after 30 minutes of consumption of anthocyanin mix- [107]. Anthocyanin, with the addition of an extra
ture from red fruit, the absorbed anthocyanins were sugar at position C-3 in the heterocyclic C-ring, has
not metabolized into the aglycones or any other meta- lower antioxidant activity than the anthocyanidin with
bolite forms in the human body [70]. a single sugar molecule [108]. Acylation of anthocyanin
The potential health benefits of anthocyanins are sum- with phenolic acid has a significant increase in antiox-
marized in Tables 2 and 3. These health benefits are idant activity [109]. Diacylation of the anthocyanin
antioxidative effects, antiangiogenesis, prevention of markedly increases the antioxidant activity but 5-gly-
CVD, anticancer, antidiabetes, improved visual health, cosylation leads to a reduction in the activity [107].
anti-obesity, antimicrobial, and nueroprotection. The A previous study reports the antioxidant activity of
mechanisms of the action of anthocyanidin and antho- malvidin-3-glucoside that was determined by metal-
cyanin in disease prevention are discussed in the next catalyzed lipid peroxidation models in comparison
section. with other antioxidants [30]. The result shows that
the quinoidal-base and pseudo-base of malvidin-3-glu-
coside significantly inhibited peroxidation of linoleate
by myoglobin compared with catechin. In the presence
Antioxidants
of hydrogen peroxide-activated myoglobin, malvidin-
The health and therapeutic effects of anthocyanin 3-glucoside had the highest antioxidant activity, fol-
are mainly contributed by its antioxidative activ- lowed by catechin, malvidin, and resveratrol. In term
ities. As reported in the literature [105], anthocya- of glycosylated anthocyanin, addition of an extra glu-
nin chalcones and quinoidal bases with a double cose to cyanidin-3-xylosyl-galactoside forms cyanidin-
bond conjugated to the keto group are efficient 3-xylosyl-glucosyl-galactoside with an ORAC value
antioxidants in scavenging free radicals. Also, the lower than the anthocyanin without addition of an
glycosylated B-ring structure of anthocyanin contri- extra sugar [107]. Acylation of malvidin-3-glucoside
butes to the high antioxidant activity, where ortho- with p-coumaric acid has antioxidant activity assessed

Table 2. Prevention of chronic diseases using plant anthocyanins.

Health benefits of anthocyanins References
Cardiovascular disease
Inhibited platelet aggregation (in vitro antithrombotic properties) [71]
Possessed vasorelaxation properties in isolated coronary artery rings of mature female pigs [72]
Decreased susceptibility to ischemia-reperfusion injury and infarct size with increased myocardial antioxidant enzyme [73]
Improved lipid profile and platelet function in healthy volunteers [74]
Anticancer effect
Suppressed cell proliferation, inflammation, and angiogenesis and induced apoptosis in esophageal tissue of rats [75]
Demonstrated significant anti-invasive potential in breast cancer cell lines (MDA-MB-231 and MCF7) [76]
Demonstrated anticancer effect on BALB/c nude mice bearing MDA-MB-453 cell xenografts and breast cancer cell lines (MCF-7, MDA-MB-231, [77]
and MDA-MB-453) by inducing apoptosis and suppressing angiogenesis
Inhibited cell migration and invasion, suppressed activation of rapidly accelerated fibrosarcoma (RAF), mitogen-activated protein kinase [78]
kinase (MEK) and c-Jun N-terminal kinase (JNK), and downregulated secretion of matrix metalloproteinase 2 (MMP2) and MMP9 of MDA-
MB-453 cells (HER2+)
Inhibited growth of human HT-29 colon cancer cells, increased expression of tumor suppression genes (p21WAF1 and p27KIP1) and [79]
decreased cyclooxygenase-2 gene expression
Reduced colonic aberrant crypt foci, colonic cellular proliferation and COX-2 mRNA expression in rats. [80]
Suppressed formation of aberrant crypt foci in colons of CF-1 mice [81]
Promoted apoptosis in benign prostatic hyperplasia rats [82]
Possessed anti-invasive effect on human hepatoma Hep3B cells and inhibited matrix metalloproteinase MMP-2 and MMP-9 gene expression [83]
Inhibited Akt-mTOR signalling thereby inducing maturation of acute myeloid leukaemia cells, besides inducing apoptotic players such as [84]
TRAIL in cancer systems
Diabetes
Ameliorated hyperglycemia and insulin sensitivity via activation of AMP-activated protein kinase in diabetic mice [85]
Improved dyslipidemia, enhanced antioxidant capacity, and prevented insulin resistance in human subjects with type 2 diabetes [86]
Alleviated glomerular angiogenesis of diabetic kidneys by attenuating the induction of VEGF and HIF-1α in studied mice [87]
Ameliorated renal apoptosis in diabetic nephropathy mice via activation of AMP-activated protein kinase (AMPK) which eventually inhibit [88]
oxidative stress and lipotoxicity.
Activated adipose tissue-derived adiponectin to defend against diabetes-related endothelial dysfunction in mice [89]
 FOOD & NUTRITION RESEARCH 9

Table 3. Other health benefits of plant anthocyanins.

Health benefits of anthocyanins References
Visual health
Improved visual function in patients with normal tension glaucoma [90]
Prevented impairment of photoreceptor cell function during retinal inflammation [91]
Decreased lens opacity together with the decreased MDA level [92]
Suppressed cell death of HLE-B3 (lens epithelial cell line) under H2O2-induced oxidative stress [93]
Prevented retinal degeneration induced by N-methyl-N-nitrosourea [94]
Increased ocular blood flows but no significant changes on intraocular pressure [95]
Anti-obesity
Improved weight gain and lipid profile on obese rats. [96]
Suppressed body weight gain and improved blood lipid profile in high-fat diet induced rats [97]
Ameliorated obesity in high-fat-fed C57BL/6 mice [98]
Up-regulate adipocytokine secretion and gene expression in isolated rat adipocytes [99]
Suppressed weight gain, fat tissue gain and other metabolic disorders [100]
Antimicrobial
Possessed antimicrobial activity through damaging and destroying the cell wall, membrane and intercellular matrix [101]
Showed antibacterial activity with the highest sensitivity to Aeromonas hydrophilia and Listeria innocua [102]
Possessed antibacterial effects towards Enterococcus faecium resistant to vancomycin, Pseudomonas aeruginosa, Staphylococcus aureus and [103]
Escherichia coli
Inhibited gram-negative bacteria but not on gram-positive bacteria [104]

by linoleic acid oxidation higher than the non-acylated Angiogenesis and development of diseases
counterpart [109].
Endothelial cells are the main cells involved in the
Anthocyanins have many other therapeutic effects
angiogenesis process. Disturbances in physiologic
in addition to their antioxidant activities. As an
angiogenesis can contribute to various human diseases,
active pharmaceutical ingredient, anthocyanin pig-
including CVDs, cancer, and diabetic complications
ment, such as delphinidin, has been patented for
such as diabetic retinopathy and nephropathy [115].
several therapeutic effects. Delphinidin is well-
Normal angiogenesis depends on the intricate balance
known for combating melanoma cells [110], as well
between angiogenic (VEGF, FGF2-fibroblast growth
as antimicrobial effects, such as curing
factor, TGF-β-transforming growth factor, and angio-
Staphylococcus aureus infection [111]. It has also
poietin) and antiangiogenic (angiostatin, endostatin,
been used as the source of antiphlogistic or immu-
and thrombospondins) factors [116].
nosuppressive active ingredients [112].
The antiangiogenic effect of anthocyanins has been
Literature shows that anthocyanins extracted from
reported by several studies. Anthocyanin-rich extracts of
plants have antioxidative properties. Pelargonidin-3-
several berries (wild blueberry, bilberry, cranberry, elder-
glucoside, cyanidin-3-glucoside, and delphinidin-3-
berry, and strawberry) significantly suppress hydrogen
glucoside isolated from Phaseolus vulgaris L. (black
peroxide and TNF-α-induced vascular endothelial growth
bean) seed coat, as well as their standard aglycones,
factor (VEGF) expression in HaCaT cells (human kerati-
have strong antioxidative activity in a liposomal sys-
nocytes) [117]. Bilberry anthocyanidins (delphinidin, cya-
tem and reduced formation of malondialdehyde by
nidin, and malvidin) are also reported to inhibit VEGF-
UVB irradiation [113]. The study also indicates that
induced tube formation in a co-culture of human umbilical
delphinidin and delphinidin-3-glucoside had the
vein endothelial cells and fibroblasts [118].
highest inhibitory effect on lipid peroxidation and
Anthocyanin-rich purple corn extract attenuates
O2•− scavenging activity. On the contrary, pelargo-
endothelial expression of VEGF and hypoxia inducible
nidin had the highest inhibitory effect on hydroxyl
factor (HIF)-1α, as well as to induce endothelial marker of
radical scavenging activity.
platelet endothelial cell adhesion molecule-1 and integrin
On the other hand, a study demonstrates that cya-
β3 induced by high glucose condition in human renal
nidin and cyanidin-3-glucoside have the highest inhi-
mesangial and endothelial cells [87]. Also, glomerular
bitory effect on copper (II)-induced low-density
angiogenesis in the diabetic kidneys of db/db mice is
lipoprotein (LDL) oxidation compared with the other
disrupted by weakening the induction of VEGF and
phenolic acids, anthocyanins, and anthocyanin agly-
HIF-1α in vivo. The purple corn extract also diminishes
cones, whereas delphinidin has intermediate efficacy
the mesangial and endothelial induction of angiopoietin
[114]. Comparing the result from both studies, the
proteins under hyperglycemic conditions. These findings
second study does not determine the efficacy of pelar-
suggest that anthocyanin-rich purple corn extract antag-
gonidin to inhibit lipid peroxidation.
onizes glomerular angiogenesis in high blood glucose
10 H. E. KHOO ET AL.

condition through disturbing the Angpt-Tie-2 ligand- and prostate cancers. The evidence from a previous
receptor system linked to the renal VEGF receptor-2 study shows that 5% whole freeze-dried black raspber-
signaling pathway. ries and the anthocyanin-rich fraction supplemented to
N-nitrosomethylbenzylamine-induced F344 rats have
chemopreventive potential, where the treatment groups
Cardiovascular health
inhibit cell proliferation, inflammation, angiogenesis,
Epidemiological studies show the relationships between and induce apoptosis in both preneoplastic and papil-
anthocyanin-rich foods and CVDs, as well as the rela- lomatous esophageal tissues [75]. Thus anthocyanins
tionship between total anthocyanin intake and risk of have chemoprophylaxis potential.
developing these cardiovascular-related diseases. Blueberry anthocyanins and anthocyanin-pyruvic
Anthocyanins also demonstrate in vitro anti-thrombo- acid adduct extracts (250 μg/ml) demonstrated anti-
tic effect [71]. The anti-thrombotic effect is supported invasive potential in both breast cancer cell lines,
by another study that anthocyanin-containing maize MDA-MB-231 and MCF7 [76]. The extracts inhib-
seed (20% seed in the diet) fed rats for eight weeks ited proliferation of cancer cell by acting as chemoin-
are less susceptible to ischemia-reperfusion injury and hibitors. The anthocyanin-pyruvic acid adduct
reduction of infarct size with increased myocardial extract has a better effect in MDA-MB-231, suggest-
antioxidant enzyme [73]. Also, Bell and Gochenaur ing an effect independent of estrogen receptors. In
[72] reveal that anthocyanin-rich extracts of choke- addition to blueberry anthocyanins, anthocyanin-rich
berry and bilberry, but not elderberry, possess vasor- extracts (50 µg monomeric anthocyanin/ml) from
elaxation properties. Moreover, there is also no chokeberry result in 60% growth inhibition of
alteration of coronary response to nitric oxide which human HT-29 colon cancer cells within 24 h expo-
is a potent vasodilator agent. sure, increase expression of tumor suppression genes
In a clinical trial, the researchers suggest that con- (p21WAF1 and p27KIP1) and a 35% decrease in the
sumption of anthocyanin-rich strawberries for one cyclooxygenase-2 gene expression. As expected, the
month improves lipid profile and platelet function in extracts have no obvious growth inhibition on nor-
healthy volunteers [74]. Nonetheless, the effects may be mal colonic cell.
attributed to the presence of non-anthocyanin com- In another study, supplementation of anthocyanin-
pounds in strawberries, such as vitamin C and phenolic rich extracts of bilberry, chokeberry, and grape (con-
compounds. Moreover, the study should have control taining 3.85 g anthocyanins per kg diet) for 14 weeks
groups for comparison. However, Curtis et al. [119] significantly reduced azoxymethane-induced aberrant
indicate the consumption of 500 mg/day of elderberry crypt foci by 26–29% in 3–4 week-old male-specific
extract for 12 weeks is ineffective in reducing the risk pathogen-free F344 rats [80]. This reduction is asso-
of CVD in healthy postmenopausal women. There is ciated with reduced cell proliferation and decreased
also no change in metabolic processing following expression of the COX-2 gene. The result also shows
12 weeks of elderberry intake compared with acute that the urinary 8-OHdG levels were similar among
intake [120]. rats fed with different diets.
Dietary anthocyanin-enriched purple-fleshed sweet
potato clone P40 significantly suppresses formation of
Anticancer
aberrant crypt foci in the colons of female CF-1 mice
Anthocyanins have been extensively studied for their coincided with a greater expression of apoptotic cas-
anticancer properties, as well as antiangiogenesis, based pase-3 in the colon mucosal epithelial cells [81]. The
on in vitro and cell culture studies, and animal models. observation suggests that anthocyanin-enriched sweet
Angiogenesis is the key for cancer development, where it potato P40 has a protective effect against colorectal
is an important step in the transition of tumors from a cancer by inducing cell-cycle arrest, anti-proliferative,
benign state to a malignant one. In cancer prevention, and through apoptotic mechanisms. Another study also
antiangiogenesis is the process that prevents formation proves that anthocyanin extract (2 and 5 mg/ml) of
of new blood vessels that supply oxygen to the tumor purple potato induces maturation of acute myeloid
cells. Several phytochemicals, including flavonoids and leukemia cells via TNF-related apoptosis-inducing
anthocyanins, are potential antiangiogenic agents. ligand [84]. Moreover, the less common anthocyanin
Anthocyanins have been extracted and isolated from source from vine was reported having anti-invasive
different plant sources for investigating their anticancer property in human hepatoma Hep3B cells in a cancer
ability on esophagus, colon, breast, liver, hematological, study [83].
 FOOD & NUTRITION RESEARCH 11

Similarly, anthocyanin-enriched black rice extract The ability of the anthocyanins to induce insulin secre-
has an anticancer effect on breast cancer cells. The tion is in the increasing order of pelargonidin-3-galacto-
extract inhibited growth of breast cancer cell lines side, cyanidin-3-glucoside, and delphinidin-3-glucoside.
MCF-7 (ER+, HER2/neu−), MDA-MB-231 (ER−, This finding demonstrates that the number of hydroxyl
HER2/neu−), and MDA-MB-453 (ER−, HER2/neu+) groups on the B-ring of anthocyanins plays a crucial role
and induces apoptosis in MDA-MB-453 cells by depo- in their ability to secrete insulin. Nevertheless, cyanidin,
larizing mitochondrial membrane potential and releas- delphinidin, pelargonidin, malvidin, and petunidin do
ing cytochrome C into the cytosol, and thus triggered not potentiate significant insulin secretion.
programmed cell death through apoptosis [77]. Oral In a clinical trial of 24 weeks involving 58 diabetic
administration of the same extract (100 mg/kg/day) to patients, the subjects in the anthocyanin group con-
BALB/c nude mice bearing MDA-MB-453 cell xeno- sumed two anthocyanin capsules (160 mg anthocya-
grafts significantly reduced tumor growth and sup- nins) twice daily purified from bilberry and
presses angiogenesis by lowering the expression of blackcurrant [86]. The results show that anthocyanin
angiogenesis factors matrix metallopeptidase-9, matrix group had a significantly lower fasting plasma glucose
metallopeptidase-2, and urokinase plasminogen activa- and insulin resistance index, as well as significantly
tor in the tumor tissue. The results from both in vitro elevated serum adiponectin and β-hydroxybutyrate
and in vivo studies suggest that anthocyanin-enriched concentrations compared to the placebo supplementa-
black rice extract exhibits anticancer capability against tion. However, the authors did not elucidate the
human breast cancer cells by inducing cell apoptosis mechanism involved for the prevention of insulin resis-
and suppressing angiogenesis. tance in the diabetic patients.
In another study, black rice anthocyanins suppress Bilberry anthocyanin has been reported to amelio-
metastasis in breast cancer cells by targeting the mito- rate hyperglycemia and insulin sensitivity via activation
gen-activated protein kinase pathway [78]. The antho- of adenosine monophosphate-activated protein kinase
cyanins inhibited migration and invasion of MDA-MB- (AMPK) in type 2 diabetic mice at skeletal muscle,
453 cells (HER2+), suppressed activation of rapidly liver, and white adipose tissue [85]. The activation of
accelerated fibrosarcoma, mitogen-activated protein AMPK causes upregulation of glucose transporter 4 in
kinase (MEK), and c-Jun N-terminal kinase (JNK), as the skeletal muscle and white adipose tissue while
well as downregulated secretion of matrix metallopro- inhibiting glucose production in the liver. AMPK acti-
teinase 2 (MMP2) and MMP9. The study suggests that vation in the liver also results in a significant reduction
black rice anthocyanins suppress metastasis in breast in liver and serum lipid content via the phosphoryla-
cancer cells by targeting the RAS/RAF/MAPK (retro- tion of acetyl-CoA carboxylase (ACC), upregulation of
virus-associated DNA sequences/rapidly accelerated peroxisome proliferator-activated receptor alpha
fibrosarcoma/mitogen-activated protein kinase) path- (PPARα), acyl-CoA oxidase, and carnitine palmitoyl-
way. Thus, it may be useful to treat patients at an transferase-1A gene expressions.
advanced cancer stage. It has been reported that a reduction in AMPK
activity leads to diabetic nephropathy, which is asso-
ciated with increased oxidative stress and lipid accu-
Antidiabetes
mulation. Supplementation of anthocyanin-rich
The antidiabetic effect of anthocyanins from plants has Seoritae extract restores AMPK activity, activates target
been widely studied. Anthocyanin-rich Cornus fruits molecules such as ACC, sterol regulatory element-
have been used in traditional Chinese prescription binding protein 1, and PPAR, and suppresses intrare-
medicines to treat diabetes [121]. Primary bioactive nal lipid accumulation in kidney tissue [88]. However,
components reported in Cornus fruits are the glyco- the authors did not examine specific contributions of
sides of cyanidin, delphinidin, and pelargonidin [98]. the bioactive compounds in the Seoritae extract to the
Jayaprakasam et al. [98] report that cyanidin-3-gluco- observed effects and the amounts of these compounds
side and delphinidin-3-glucoside effectively aided insu- incorporated into the kidney. It is unsure if the target
lin secretion from rodent pancreatic β-cells (INS-1 832/ of anthocyanins is only AMPK or adiponectin. During
13) in vitro compared with the other anthocyanins and the onsets of diabetic microangiopathic, microvascular
anthocyanidins studied. permeability and the number of leucocytes sticking to
Another study demonstrates that pelargonidin and the venular endothelium are increased [123]. In db/db
pelargonidin-3-galactoside caused a 1.4-fold increase in mice, cyanidin-3-glucoside (2 g/kg diet) increases adi-
insulin secretion at 4 mM glucose concentration repre- ponectin secretion from adipose tissue, thus it protects
sentative of the normal glucose level in human [122]. the mice against diabetes-related endothelial
12 H. E. KHOO ET AL.

dysfunction [89]. The study also shows that cyanidin- regulatory element binding protein-1 mRNA level in
3-glucoside supplementation for eight weeks resulted the white adipose tissue. These downregulations may
in a noticeable improvement in endothelium-depen- contribute to a low triacylglycerol accumulation in
dent relaxation of aorta of the mice. white adipose tissue.
Obesity is strongly associated with adipocyte dys-
function. Therefore, regulation of protein secretion
Visual health
from adipocyte or the adipocyte-specific gene expres-
Anthocyanin pigments are important nutraceuticals in sion is one of the most important targets for preven-
maintaining good vision. Anthocyanin-rich berries are tion of obesity. Tsuda and his research team further
traditionally known for the goodness to eyes and are investigated the potency of anthocyanins, particularly
often associated with night vision. Most of the berries cyanidin and cyanidin-3-glucoside on isolated rat adi-
have high anthocyanins content. Oral administration of pocytes for anti-obesity effect. He demonstrates that
bilberry extract (contained about 39% anthocyanins) to the adipocytes treated with anthocyanins have
six weeks old C57BL/6 mice has been shown to prevent increased adiponectin and leptin secretions and upre-
impairment of photoreceptor cell function during ret- gulated adipocyte-specific gene expression without
inal inflammation [91]. In another study, 132 patients activation of PPARγ in the isolated rat adipocytes
with normal tension glaucoma were supplemented with [99]. Gene expression of adiponectin is also upregu-
two anthocyanins capsules (60.0 mg anthocyanin in lated in white adipose tissue of the anthocyanin-treated
each capsule) from bilberry daily and have improved mice. The increased phosphorylation of AMPK may be
visual function, based on the Humphrey visual field associated with these changes, and the monopho-
test and minimal angle of resolution best-corrected sphate/adenosine triphosphate ratio is significantly
visual acuity assessment [90]. decreased by the administration of anthocyanins.
Some other berries demonstrate a protective effect As previously reported by Tsuda et al., adipocyte
for eyesight. Blackcurrant anthocyanin supplementa- gene expression is not thoroughly studied. A further
tion (50 mg/day) for 24 months increased ocular examination of gene expression profile in isolated rat
blood flow in 19 patients with open-angle glaucoma, adipocytes treated with anthocyanins (100 nM cyani-
however, there were no significant changes in the din-3-glucoside or cyanidin) has been performed in
intraocular pressure [95]. Supplementation of antho- vitro [125]. Within 24 h, a total of 633 genes and 427
cyanins (50 mg/kg body weight) from the seed coat of genes were upregulated (1.5-fold) by the treatment of
black soybean to N-methyl-N-nitrosourea-induced ret- adipocytes with cyanidin-3-glucoside and cyanidin,
inal degenerative rats also prevents retinal degeneration respectively. The upregulated genes include lipid meta-
[94], and also suppresses human lens epithelial cell bolism and signal transduction-related genes. However,
death under hydrogen peroxide-induced oxidative the altered genes are partially different when compar-
stress by 50–200 μg/ml of the extract [93]. ing the cyanidin-3-glucoside and cyanidin treated
Anthocyanin also predominates around 70% in purple groups. They also report that treatment of adipocytes
corn seed [124], where purple corn seed extract with cyanidin-3-glucoside and cyanidin upregulated
decreases lens opacity together with lower malonalde- hormone sensitive lipase and enhanced lipolytic activ-
hyde levels [92]. ity based on the microarray data. Even though the
findings have identified new responsive genes with
potentially important functions in adipocytes related
Anti-obesity effect
to obesity, additional investigation is needed. In vivo
Anthocyanidin and anthocyanin pigments possess anti- adipocytes are not likely to be exposed to the antho-
obesity properties. Based on a previous study, obese cyanidin due to its instability in the culture.
mice fed a diet rich in cyanidin-3-glucoside from pur- Another study found that the ameliorated obese
ple corn for 12 weeks have reduced body weight, as mice (C57BL/6) fed with Cornelian cherries (Cornus
well as decreases in white and brown adipose tissue mas) containing anthocyanins (1 g/kg of high fat diet)
weights [100]. The study demonstrates that hypergly- for eight weeks had a 24% decrease in weight gain and
cemia, hyperinsulinemia, hyperleptinemia, and an decreased lipid accumulation in the liver, as well as a
increase in the tumor necrosis factor (TNF-a) mRNA significant decrease in liver triacylglycerol concentra-
level occurred in the obese rats are normalized when tion, independent of food intake [126]. The diet con-
treated with purple corn diet. The purple corn also taining a mixture of delphinidin, cyanidin, and
suppresses mRNA levels of enzymes involved in fatty pelargonidin-3-O-galactosides. On the contrary, con-
acid and triacylglycerol synthesis and lowered sterol sumption of whole blueberry powder and isolated
 FOOD & NUTRITION RESEARCH 13

anthocyanins from blueberry and strawberry yields a hydrophilic compounds [131]. These antimicrobial
mixed result. In addition, high-fat diet mice fed with activities of anthocyanin-containing extracts are pos-
whole blueberry powder have increased body weight sibly due to the multiple mechanisms and synergistic
and adiposity relative to high-fat-fed controls [127]. effects of various phytochemicals in the extracts,
Inversely, the study shows that the obese mice fed including anthocyanins, weak organic acids, phenolic
with isolated anthocyanins from the fruits reduced acids, and their mixtures of different chemical forms
weight gain and body fat, but the differences were not [132]. Thus, the antimicrobial effect of chemically
always statistically significant. The authors also tested complex compounds instead of solely anthocyanins
the purified anthocyanins and blueberry juice for the should be extensively analyzed. Also, anthocyanins in
ability to prevent obesity by preparing a dose of purple, red, and blue-colored fruits and vegetables are
0.2 mg/ml anthocyanin in drinking water (0.49 mg/ the main bioactives in preventing microbial infection
mouse/day). The finding shows that consumption of by several mechanisms.
the purified anthocyanins suppressed the rate of fat
deposition. Also, consumption of blueberry juice
Neuroprotective effect of anthocyanins
(2.8 ml/mouse/day; 5.3 mg of anthocyanin/mouse/
day) was not as effective as the purified anthocyanins The term, ‘neuroprotection’, has been defined as the
in preventing deposition of fat in the body. Moreover, protection of nerve cells from oxidative injury and
lower serum leptin concentrations had been consis- neurotoxicity, which interferes with the ischemic cas-
tently observed in the purified blueberry anthocyanins cade. A neuroprotective agent is a drug or natural
(1.0 mg/ml) fed to obese mice for 72 days, which compound that prevents the nervous system from sec-
reduces the development of obesity [128]. ondary injuries. The neuroprotective effects of antho-
cyanins have been evaluated based on the in vitro and
in vivo studies. Most of the in vitro studies are per-
Antimicrobial
formed by applying cell cultures, whereas in vivo stu-
Polyphenolic compounds including anthocyanins pos- dies are carried out based on animal models.
sess antimicrobial activity against a wide range of The neuroprotective findings from selected studies
microorganisms, especially in inhibiting the growth of are reported in this review. An in vitro study [133]
food-borne pathogens [129]. Anthocyanins exhibit shows the neuroprotective effect of cyanidin-3-gluco-
antimicrobial activity through several mechanisms, side and its aglycone against hydrogen peroxide-
such as induced cell damage by destroying the cell induced oxidative stress in human neuronal cells (SH-
wall, membrane, and intercellular matrix [101]. SY5Y). The results demonstrate that SH-SY5Y cells
Based on a previous study, maqui berry extracts had pretreated with 100 µM cyanidin and cyanidin-3-glu-
antibacterial activity with the highest sensitivity to coside significantly increased total antioxidant activity
Aeromonas hydrophilia and Listeria innocua [102]. of membrane and cytosolic fraction from the cells;
These bacteria are commonly associated with refriger- cyanidin also significantly increased the percentages
ated foods as indicators of pathogenic microorganisms of mitochondrial functioning and inhibited DNA frag-
or as spoilage microorganisms [130]. Côté et al. [103] mentation induced by hydrogen peroxide.
report that cranberry extract had antibacterial activity Based on a previous study, cyanidin-3-glucoside
towards Enterococcus faecium resistant to vancomycin, (2 mg/kg body weight) isolated from Prunus cerasus
Pseudomonas aeruginosa, Staphylococcus aureus, and fruit inhibited apoptosis-inducing factor from mito-
Escherichia coli. The antibacterial activity of cranberry chondria under oxidative stress but did not block the
extract is not based on its low pH, but it is believed due release of cytochrome c against permanent middle cer-
to the other specific bioactive components, such as ebral artery occlusion in the cortical neurons isolated
anthocyanins and flavonols in cranberry extracts after from adult mice brain [134]. Besides the reported find-
pH adjusted to 7. ings, the cyanidin-3-glucoside treated mice had brain
Anthocyanin-rich extracts, such as blueberry, rasp- superoxide level lower than the control (0.9% normal
berry, blackcurrant, and strawberry extracts, inhibit saline), as well as with better neurological test scores.
Gram-negative bacteria but not Gram-positive bac- In another study, male Sprague-Dawley rats with
teria [104]. This variation may be due to the different traumatic spinal cord injury that received 400 mg/kg
structures of cell wall between Gram-negative and body weight of cyanidin-3-glucoside had a significantly
Gram-positive bacteria, in which the outer membrane improved blood-brain barrier score by 16.7%, platform
of Gram-negative bacteria acts as a preventive barrier hang by 40.0%, and hind foot bar grab by 30.8% com-
against hydrophobic compounds but not on pared to vehicle treated control, as well as significant
14 H. E. KHOO ET AL.

reductions in superoxide level of the spinal cords and risk of several diseases that can be shown by direct and
lesion volume in the lesion periphery, and a significant indirect pathways. Direct pathway is that the colored
increased in motor neuron cell numbers of the anterior compounds directly reduce the risk of several chronic
horn in lesion periphery [135]. The data from a mouse diseases through scavenging free radicals and thus redu-
model of late pregnancy reveals that intraperitoneal cing oxidative stress. The indirect pathways involve
injection of cyanidin-3-glucoside blocks ethanol- downregulation of cell proliferation and apoptosis
mediated glycogen synthase kinase-3β by inducing through reduction of oxidative stress and lipid peroxida-
phosphorylation at serine 9 and reduces the phosphor- tion. It is commonly known that anthocyanins are the
ylation at tyrosine 216 [136]. The compound also ame- strong antioxidants that effectively scavenge free radicals.
liorates ethanol-induced oxidative stress by inhibiting Anthocyanins reduce the risk of CVD through improv-
expression of malondialdehyde (MDA). The study con- ing blood lipid profile and biomarkers. A reduction in
cludes that cyanidin-3-glucoside prevents neurotoxicity certain blood biomarkers is known to prevent CVDs.
of ethanol. Similar to many other phenolic compounds, anthocya-
As reported in the literature, cyanidin-3-O-ß-d-gluco- nins inhibit cancer cell proliferation via several pathways.
pyranoside isolated from mulberry extract had a neuro- One of the well-known mechanisms of action is the
protective effect on PC12 pheochromocytoma cells downregulation of cyclooxygenase (COX) enzyme activ-
through inhibiting cerebral ischemic damage induced ity. These enzymes catalyze the formation of leuko-
by oxygen-glucose deprivation when the cells were trienes, prostacyclins, prostaglandins (PGs), and
exposed to hydrogen peroxide (150 µM) for 24 h [137]. thromboxanes [139]. Downregulation of COX enzymes,
The researchers also developed a mouse-brain-injury including COX-1 and COX-2, reverses cell proliferation
model of transient middle-cerebral artery occlusion, and thus reduces the risk of cancer. Anthocyanins also
where the mice were supplemented with cyanidin-3-O- inhibited tumor growth by blocking activation of the
β-d-glucopyranoside and mulberry fruit extract. The mitogen-activated protein kinase pathway. Moreover,
result demonstrates a reduction in the infarct volume of the most commonly known pathways are cytokine sig-
the brain by 18% and 26%, respectively, and both supple- naling pathways. The analysis of structure-activity rela-
mented groups had a lesser number of myeloperoxidase- tionships among flavonoids suggests that 4-
positive cells than the ischemic control group in the hydroxylations at positions 5, 7, 31, and 41, together
striatum and cortex of the injured brain. with a bond at C2–C3, and the B-ring attaching at the
Based on the previous findings, most of the stu- C2 position, seem necessary for the highest expression of
dies report the neuroprotective effects of cyanidin monocyte chemoattractant protein 1 (MCP-1) [140].
and its glycosides. Limited study has been done to
determine the neuroprotective benefits of other
Free-radical scavenging pathway
anthocyanidins and anthocyanins. Kim et al. [138]
show the neuroprotective effect of three major Free radicals are generated during oxidative stress in
anthocyanins (a mixture of cyanidin-3-glucoside, the cellular system. Reactive oxygen species (ROS) and
delphinidin-3-glucoside, and petunidin-3-glucoside) reactive nitrogen species (RNS) are the typical free
isolated from black soybean, against hydrogen per- radicals that are produced during oxidative stress.
oxide-induced cell death. They conclude that the ROS and RNS are generated in several cellular systems
human brain neuroblastoma SK-N-SH cells treated in the human body. ROS is typically produced in
with the purified anthocyanin mixture (1–25 μg/ml) cytosol, mitochondrial, peroxisomes, endoplasmic reti-
had a significant reduction in intracellular ROS level culum, plasma membrane, and lysosomes, whereas
in a dose-dependent manner. The anthocyanins also RNS is produced from amino acid metabolism [141].
inhibited ROS-dependent activation of apoptosis sig- The most commonly known ROS signaling is
nal-regulating kinase 1 (ASK1)–JNK/p38 pathways, through the respiratory chain. It involves an electron-
stimulated expression of heme oxygenase 1, and transfer reaction pathway, where superoxide dismutase
upregulated sialidase 1 (also known as Neu1) gene (SOD) enzyme produces hydrogen peroxide (H2O2) in
expression. Based on this evidence, anthocyanins mitochondrial. At the Complexes I and III in mitochon-
obtained from plants have a neuroprotective effect. drial, hydroxyl radical (•OH) is generated via a Fenton
reaction (H2O2 + Fe2+→ •OH + OH− + Fe3+). There are
other pathways involved in the production of ROS
Mechanisms of action in disease prevention
(O2•−), such as through a α-ketoglutarate dehydrogenase
Anthocyanins are the good antioxidants for preventing complex and by several oxidoreductases in the mito-
or reducing the risk of disease. Anthocyanins reduce the chondrial [141]. A similar mechanism of action also
 FOOD & NUTRITION RESEARCH 15

occurs in peroxisomes and lysosomes, which involves antioxidant biomarkers. Oxidative free radicals also
metabolism of H2O2. initiate inflammatory responses of vascular endothelial
Another pathway for ROS generation is the xeno- cells and upregulate cell adhesion molecules and
biotic metabolism in the endoplasmic reticulum. In chemokines.
the reticulum, oxygen (O2) initiates a reaction with The results obtained from a double-blind clinical
lipophilic substrates (such as fatty acids, FH) in the trial of 150 hypercholesterolemic subjects aged
presence of a reducing agent-RH2 (FH + O2 + RH2 40–65 years old show that consumption of anthocya-
→ AOH + R + H2O). The reaction continues and nins (total intake of 320 mg anthocyanins per day) for
thus produced ROS in the microsomes. In both the 24 weeks significantly reduced the serum high sensitiv-
endoplasmic reticulum and plasma membrane, lipid ity C-reactive protein (−21.6%), soluble vascular cell
peroxidation occurs via NADPH (nicotinamide ade- adhesion molecule-1 (−12.3%), and plasma IL-1β
nine dinucleotide phosphate) formation pathways. (−12.8%) compared to the placebo (−2.5%, 0.4%, and
When NADPH oxidase is activated, a large amount −1.3%, respectively) [146]. Another double-blind, ran-
of O2•− and H2O2 are generated. Therefore, the ROS domized, placebo-controlled trial also shows consump-
produced is known to cause cell proliferation in a tion of anthocyanins (160 mg anthocyanins twice daily)
cellular system. for 12 weeks significantly increased the serum HDL-
Anthocyanins, as the well-known antioxidants and cholesterol level (13.7%) and decreased the LDL-cho-
free radical scavengers, are able to act as reducing agents lesterol level (13.6%) compared to the placebo group
in the electron-transfer reaction pathway. The antioxida- (2.8% and −0.6%, respectively) [147]. In addition, a
tive compounds are able to donate electrons to the free meta-analysis concludes that anthocyanin supplemen-
radicals with unpaired electrons [142]. Anthocyanins also tation to patients with dyslipidemia gave a significant
scavenge free radicals through two pathways that have reduction in serum TC, triglyceride, and LDL-choles-
been hypothesized in the past decades. The first pathway terol levels, as well as a significant increased in the
is the attack of hydroxyl group(s) of the B-ring of HDL-cholesterol level [148].
theanthocyanin structure and the second is the attack of
oxonium ion on the C-ring. Anthocyanins are some of
Cyclooxygenase (COX) pathway
the strongest antioxidants due to the free radical scaven-
ging abilities via both pathways. COX-1 is essential for formation of thromboxane in
Literature supports the fact that the number of blood platelets and maintaining integrity of gastroin-
hydroxyl groups on the B-ring of the anthocyanin testinal epithelium. It is expressed in most of the tis-
structure affects the scavenging activity of the antho- sues. It is known for involvment in cell signaling,
cyanin molecules [143]. The number of hydroxyl regulating angiogenesis in endothelial cells, and main-
groups is positively associated with the scavenging taining tissue homeostasis. COX-2, also known as pros-
activity. Nonetheless, no study reports on the mechan- taglandin-endoperoxide synthase 2, is recognized to be
ism of the positive charge at the oxygen atom of the the essential enzyme that is involved in the conversion
C-ring of the anthocyanin structure for scavenging of of arachidonic acid (ARA) to PGH2 (ARA → PGG2 →
free radicals. It has also been hypothesized that the PGH2). The biosynthetic pathway of PGs has been
superoxide O2o- radical favors the oxonium ion of reported by Ricciotti and FitzGerald [149] in their
anthocyanin [144]. review article.
COX-1 and COX-2 initiate the conversion of ARA
into PGH2, where it serves as a substrate for producing
Biomarkers and mechanism for cardiovascular
a series of specific isomerases. As reported in the lit-
diseases
erature, PGE2, PGI2, PGD2, and PGF2α are the four
Oxidative damage to a cardiovascular system is typi- principal bioactive prostaglandins studied. PGE2 and
cally caused by ROS and RNS. During the development PGI2 are typically expressed in the vascular smooth
of CVD, oxidative stress causes vascular inflammation. muscle cells, platelets, as well as brain and kidney
Vascular inflammation alters the levels of cellular total cells. PGI2 is the key prostanoid that regulates cardio-
cholesterol (TC), LDL, high-density lipoprotein (HDL), vascular homeostasis [149]. Oxidative stress increases
and very low-density lipoprotein (VLDL), as well as expression of PGI2 in the vascular cells. PGI2, together
plasma malonaldehyde and other plasma enzymes with PGE2, mediates vasodilation of vascular system.
(SOD, catalase, and glutathione peroxidase) levels COX-2 is overexpressed in benign polyps and adeno-
[145]. These molecules are the prognostic markers for carcinomas [150]. The role and mechanism of COX-2
CVD, also called the plasma lipid profile and in cell proliferation and cell death have been clearly
16 H. E. KHOO ET AL.

explained in a review article [151]. The authors of this cytokine IL-1 also activates p38 MAPKs during the
review article explain the roles of COX-2 in the preven- inflammatory process [154].
tion of cancer, which involve cell signaling and regula- An in vivo study shows that lipopolysaccharide-
tion of cell proliferation and apoptosis. They also discuss induced mice fed with 10% blueberries have reduced
the role of PGE2 in inhibiting apoptosis in several in expressions of protein and mRNA of TNF-α and IL-6
vitro models. in the serum compared to the control [158]. A rando-
mized control trial also shows that hypercholesterole-
mic subjects who consumed a purified anthocyanin
Mitogen-activated protein kinase pathway mixture (320 mg/day) for 24 weeks had a significantly
Mitogen-activated protein kinases (MAPKs) are the lower level of plasma IL-1β than the placebo [146]. On
protein kinases involved in cell survival, such as cell the contrary, the control trial reveals no significant
proliferation, differentiation, migration, and apoptosis reduction in TNF-α level between the treatment and
[152]. The types of known MAPKs are extracellular placebo at the end of the study (week-24). Another
signal-regulated kinase 1 and 2 (ERK 1/2), ERK 5, study also indicates that treatment of human monocy-
JNK 1–3, and p38 protein isoforms (p38α, β, γ, and tic THP-1 cells with 10 mg/ml anthocyanin-containing
δ) [153]. Among the MAPKs, p38 MAPKs regulate the bilberry extract (25% anthocyanin content) signifi-
expression of many cytokines [154]. cantly lowered IFN-γ-induced (100 ng/ml) expressions
Among the six typical anthocyanins, peonidin-3- of MCP-1, IL-6, TNF-α, intercellular adhesion mole-
glucoside is reported to downregulate ERK 1/2 expres- cule 1 (ICAM-1), and T-cell-specific T-box transcrip-
sion in the H1299 cell line [155]. Anthocyanin-rich tion factor (T-bet) [159].
pomegranate extract also had a positive effect on UV-
B-mediated phosphorylation of MAPKs pathway in
normal human epidermal keratinocytes. The study Conclusions
reveals that the pomegranate extract (20 µg/mL) inhib-
ited UV-B–mediated phosphorylation of MAPK (ERK Anthocyanins are colored pigments in plants that possess
l/2, p38 protein, and JNK 1/2) in a time-dependent several health benefits. These colored pigments appear
manner [156]. Anthocyanin extract obtained from red in acidic condition and show a blue hue in alkaline
Vitis coignetiae Pulliat has anti-tumor activity. The solution. Acylated and copigmentated anthocyanidins
extract (≤60 µg/ml), which contains delphinidin-3,5- have higher heat stability, thus maintain the structure
diglucoside, cyanidin-3,5-diglucoside, petunidin-3,5- even in different pH conditions. Anthocyanins are the
diglucoside, delphinidin-3-glucoside, malvidin-3,5- value-added colorants that can be used for preventing
diglucoside, peonidin-3,5-diglucoside, cyanidin-3-glu- several diseases, including CVDs, cancers, diabetes,
coside, petunidin-3-glucoside, peonidin-3-glucoside, some metabolic diseases, and microbial infection. These
and malvidin-3-glucoside, induces apoptosis of HCT- compounds also improve visual ability and have neuro-
116 cells through increasing phosphorylation of p38- protective effect. Several mechanisms of action are
MAPK and ERK, as well as suppressing phosphoryla- reported for the anthocyanidins and anthocyanins in
tion of Akt (protein kinase B) and JNK [157]. prevention of these diseases. In a nutshell, free-radical
scavenging, changes in blood biomarkers, COX and
MAPKs pathways, as well as inflammatory cytokines
Inflammatory cytokines signaling signaling are the typical mechanisms of action of these
Chronic inflammation is linked to progression of a dis- colored pigments in prevention of diseases.
ease that is characterized by excessive production of cyto-
kines, changes in the pattern of cellular signaling, and
infiltration of inflammatory cells. Similar to most of the Disclosure statement
flavonoids, anthocyanins reduce inflammation through
No potential conflict of interest was reported by the authors.
several mechanisms to attenuate and to prevent inflam-
matory responses. The inflammatory cells produce sev-
eral cytokines, including IL-1, IL-6, and TNF-α, that ORCID
change the size and number of the cells. During the
Hock Eng Khoo http://orcid.org/0000-0002-7686-6092
inflammation process, these cytokines activate hypotha- Azrina Azlan http://orcid.org/0000-0002-5373-0985
lamic-pituitary-adrenal that acts on the release of gluco- Sou Teng Tang http://orcid.org/0000-0002-1733-4352
corticoids from the adrenal cortex. The inflammatory See Meng Lim http://orcid.org/0000-0002-8983-9245
 FOOD & NUTRITION RESEARCH 17

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