JOURNAL OF FUNCTIONAL FOODS x x x ( 2 0 1 3 ) x x x –x x x

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Green tea extract: Chemistry, antioxidant properties

and food applications – A review

S.P.J. Namal Senanayake*

DuPont Nutrition & Health, Danisco USA Inc., Four New Century Parkway, New Century, Kansas 66031, United States

A R T I C L E I N F O A B S T R A C T

Article history: Green tea is one of the most popular and extensively used dietary supplement in the United
Received 20 May 2013 States. Diverse health claims have made for green tea as a trendy ingredient in the growing
Received in revised form market for nutraceuticals and functional foods. Green tea extract contains several polyphe-
16 August 2013 nolic components with antioxidant properties, but the predominant active components are
Accepted 22 August 2013 the flavanol monomers known as catechins, where epigallocatechin-3-gallate and epicate-
Available online xxxx chin-3-gallate are the most effective antioxidant compounds. Additional active compo-
nents of green tea extract include the other catechins such as epicatechin and
Keywords: epigallocatechin. Among these, epigallocatechin-3-gallate is the most bioactive and the
Green tea extract most scrutinized one. Green tea polyphenols are also responsible for distinctive aroma,
Antioxidant color and taste. Green tea extract can also be used in lipid-bearing foods to delay lipid oxi-
Catechins
dation and to enhance the shelf-life of various food products. This review outlines the
Flavour constituents
chemistry, flavour components, antioxidant mechanism, regulatory status, food applica-
Lipid oxidation
tions, and stability of green tea extract in food.
Food application
Ó 2013 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
2. Chemistry. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
3. Flavour constituents. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
4. Antioxidant mechanism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
5. Regulatory status . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
6. Method of incorporation into food . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
7. Application in food products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
8. Stability tests . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
8.1. Peroxide value (PV) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
8.2. Conjugated dienes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
8.3. Thiobarbituric acid (TBA) value. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
8.4. Oil Stability Index (OSI) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
8.5. Oxipresä . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
8.6. Schaal oven storage test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00

* Tel.: +1 800 255 6837; fax: +1 913 764 0854.

E-mail address: namal.senanayake@dupont.com.
1756-4646/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jff.2013.08.011

Please cite this article in press as: Namal Senanayake, S.P.J., Green tea extract: Chemistry, antioxidant properties and food applications – A re-
view, Journal of Functional Foods (2013), http://dx.doi.org/10.1016/j.jff.2013.08.011
2 JOURNAL OF FUNCTIONAL FOODS
8.7. Sensory evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
9. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 00
1. Introduction
Tea, derived from Camellia sinensis L., is one of the most widely
consumed beverages in the world. Tea can be categorized into
three main types, depending on the level of oxidation, as
green tea, oolong tea and black tea (Chan, Soh, Tie, & Law,
2011; Velayutham, Babu, & Liu, 2008). Green tea is an ever-
green plant that grows primarily in tropical and temperate re-
gions of Asia which mainly include China, India, Sri Lanka
and Japan. It is also cultivated in several African and South-
American countries. Two primary varieties of C. sinensis are
Camellia sinensis sinensis and Camellia sinensis assamica. The
sinensis plant strain is originated from China. This strain pro-
duces green, white, black and oolong teas. On the contrary,
the assamica plant strain primarily is inhabitant to the Assam
region in Northern India. Due to enormous yields of this spe-
cific strain, it is the favored plant grown in India, Sri Lanka
and some African countries. The leaves of assamica strain
are typically used for producing black, oolong and pu’erh teas.
Green tea is a small shrub that can expand up to 30 feet high,
but is customarily trimmed to 2–5 feet when cultivated for its
leaves. The leaves are naturally murky green and glossy with
notched edges and are 2–5 cm broad and 4–15 cm long. The
flowers are white and contain bright yellow stamens. These
blossoms appear individually or as clusters. The fruits have
hard green shells with round, brown-colored seeds. These
seeds can be used to produce tea oil. Typically, flowering is
prevented during cultivation by harvesting the leaves. The
immature, light-green leaves are preferably harvested for
tea production. Mature leaves are deeper green in color than
the young leaves. Different leaf ages produce varying tea
qualities as their chemical compositions are different. Typi-
cally, the buds (tips) and the first two to three leaves are har-
vested by hand picking for processing. This process is
generally repeated every one to two weeks. Green, white,
black and oolong teas all originate from the leaves of C. sinen-
sis plant. However, the different types of tea are classified
according to the degree of fermentation that takes place dur-
ing processing: both white tea and green tea being unfer-
mented; oolong tea semi-fermented and black tea fully
fermented (Chan et al., 2011). These basic types of tea have
different quality characteristics, including appearance, aro-
ma, taste, and color. The manufacturing process of tea is
designed to either preclude, or permit tea polyphenolic com-
pounds to be oxidized by naturally occurring polyphenol
oxidase in the tea leaves. Green tea is produced by inactivat-
ing the heat-labile enzyme polyphenol oxidase in the fresh
leaves by either applying heat or steam, which prevents the
enzymatic oxidation of catechins, the most abundant
flavonoid compounds present in green tea extracts (Velayu-
tham et al., 2008). The tea leaves are then rolled, dried and
packaged.
Please cite this article in press as: Namal Senanayake, S.P.J., Green tea extract: Chemistry, antioxidant properties and food applications – A re-
view, Journal of Functional Foods (2013), http://dx.doi.org/10.1016/j.jff.2013.08.011
x x x ( 2 0 1 3 ) x x x –x x x
2. Chemistry
The chemical composition of tea leaves has been well docu-
mented. The main constituents of tea leaves are polyphenols
(Balentine, Wiseman, & Bouwens, 1997). The fresh tea leaves
contain caffeine (approximately 3.5% of the total dry weight),
theobromine (0.15–0.2%), theophylline (0.02–0.04%) and other
methylxanthines, lignin (6.5%), organic acids (1.5%), chloro-
phyll (0.5%) and other pigments, theanine (4%) and free amino
acids (1–5.5%), and numerous flavour compounds (Graham,
1992). In addition, a wide variety of other components exists,
including, flavones, phenolic acids and depsides, carbohy-
drates, alkaloids, minerals, vitamins and enzymes (Chaturve-
dula & Prakash, 2011). Tea also contains flavonols, mainly
quercetin, kaempferol, myricetin, and their glycosides. The
most favorable effects of green tea are accredited to the green
tea polyphenols, predominantly the catechins, which make
up, 25–35% of the dry weight of green tea leaves (Abdel-Rah-
man et al., 2011; Balentine et al., 1997; Graham, 1992; Zaveri,
2006). The tea catechins belong to the family of flavonoids
(Yilmaz, 2006) and possess two benzene rings referred to as
the A- and B-rings. In addition, the catechin molecules con-
tain a dihydropyran heterocycle (the C-ring) that has a hydro-
xyl group on carbon 3. Moreover, the A-ring is similar to a
resorcinol moiety whereas the B-ring is similar to a catechol
moiety. The catechin molecule has two chiral centers on car-
bons 2 and 3. Hence, it has four diastereoisomers with two of
the isomers are in trans configuration, and the other two are
in cis configuration. The trans and cis isomers are referred to
as the catechin and epicatechin, respectively. These chemical
structures appear to be important for the antioxidant activi-
ties of tea polyphenols, including an ortho-3 0 4 0 -dihydroxyl
group or 3 0 4 0 5 0 -trihydroxyl group in the B-ring, a gallate group
located at the 3 position of the C-ring, and hydroxyl groups at
the 5 and 7 positions of the A-ring (Rice-Evans, Miller, & Pa-
ganga, 1996). Many structure–activity relationship investiga-
tions have been performed on the antioxidant activity of
flavonoids, including tea catechins (Farkas, Jakus, & Héberger,
2004; Guo et al., 1999; Harborne & Williams, 2000; Heim,
Tagliaferro, & Bobilya, 2002; Rice-Evans et al., 1996). According
to these studies, the antioxidant activity of flavonoids de-
pends substantially on the number and position of hydroxyl
groups in the molecule (Farkas et al., 2004). In addition, sev-
eral structural elements such as o-dihydroxyl catechol struc-
ture in the B-ring, the presence of unsaturation and 4-oxo
group in the C-ring are also presumed to increase the antiox-
idant activity of flavonoids. The 2,3-double bond in the C-ring
along with 4-oxo function in the C-ring facilitates electron
delocalization from the B-ring. Moreover, hydroxyl groups at
positions 3 and 5 providing hydrogen bonding to the 4-oxo
group in the C-ring is another structural feature attributed
to the antioxidant activity of flavonoids.
 JOURNAL OF FUNCTIONAL FOODS
Fig. 1 – Chemical structures of major catechins found in
green tea extract.
Extracts of immature tea leaves are rich in flavanols and
their gallic acid derivatives, namely, (+)-catechin, ()-epicate-
chin, (+)-gallocatechin, ()-epicatechin gallate, ()-epigalloca-
techin, and ()-epigallocatechin gallate (Wanasundara &
Shahidi, 2005). In addition, they contain a range of natural
green tea flavour components such as terpenes, oxygenated
terpenes, sesquiterpenes, and organic acids. As exemplified
in Fig. 1, catechins are characterized by various hydroxyl
groups on the A- and B-rings (Velayutham et al., 2008). Epicat-
echin has an ortho-dihydroxyl group in the B-ring at carbons
3 0 and 4 0 and a hydroxyl group at carbon 3 on the C-ring
(Fig. 1). Epigallocatechin differs from epicatechin in that it
has a trihydroxyl group at carbons 3 0 , 4 0 , and 5 0 on the B-ring
(Yilmaz, 2006). Epicatechin gallate differs from epicatechin in
its gallate moiety positioned at carbon 3 of the C-ring (Yilmaz,
2006). However, epigallocatechin gallate is composed of tri-
hydroxyl groups on the B-ring and a gallate moiety located
at carbon 3 of the C-ring (Yilmaz, 2006). In epigallocatechin
gallate, these trihydroxyl groups are located at carbons 3 0 , 4 0
and 5 0 on the B-ring. Yilmaz (2006) has reviewed the chemis-
try and application perspectives of green tea, especially in
relation to using their catechin components. The relative con-
tent of green tea catechins depends on how the leaves are
processed prior to drying. Fermentation and heating of tea
leaves during the manufacturing process may cause polymer-
ization of monopolyphenolic catechins, leading to conforma-
tional changes and thus modifying their properties (Cabrera,
Artacho, & Giménez, 2006).
Green tea contains significantly higher amounts of poly-
phenols as compared to black or oolong tea, owing to differ-
ences in the processing of tea leaves following harvest. For
green tea, freshly harvested tea leaves are steamed at high
temperatures and dried to deactivate the polyphenol oxidases
and, as a result, preventing the oxidation of catechins and
maintaining the polyphenols in their monomeric forms. In
addition to preventing polyphenol oxidation, the steaming
process also protects against enzymatic degradation of vita-
mins. Through separation, purification, concentration and
the drying processes, highly concentrated green tea catechin
extract with low flavour intensity can be produced. Black tea,
Please cite this article in press as: Namal Senanayake, S.P.J., Green tea extract: Chemistry, antioxidant properties and food applications – A re-
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x x x ( 2 0 1 3 ) x x x –x x x 3
on the other hand, is produced by extended fermentation of
tea leaves, which results in, the development of polymeric
compounds including thearubigins and theaflavins (Zaveri,
2006). Black tea contains predominantly gallates of epicate-
chin. Oolong tea, a partially fermented product, contains a
mixture of the monomeric polyphenols and high-molecular-
weight theaflavins (Graham, 1992; Zaveri, 2006). All three vari-
eties of tea contain significant quantities of caffeine (3–6%)
which is unaffected by different processing conditions (Chu,
1997; Zaveri, 2006). Furthermore, all three varieties of tea con-
tain ()-epicatechin, ()-epigallocatechin, ()-epicatechin gal-
late and ()-epigallocatechin gallate, with the exception of
catechin (Khokhar, Venema, Hollman, Dekker, & Jonge,
1997). Epicatechin gallate and epigallocatechin gallate are
considered to be the main catechins found in black tea (Oban-
da, Owuor, & Mang’oka, 2001; Yilmaz, 2006). Epigallocatechin
gallate is the most copious catechin component present in
the leaves of green, black and oolong teas (Graham, 1992). In
general, the contents of epigallocatechin gallate in green
and oolong teas range from 22 to 53 mg/g of commercial teas
(Zuo, Chen, & Deng, 2002). However, epigallocatechin gallate
content in black teas is about 4.0 mg/g of commercial teas
(Zuo et al., 2002).
The stability of green tea catechins depends on the pH and
temperature. In acidic solutions (pH < 4), green tea catechins
exhibit exceptional stability; in alkaline solutions (pH > 8),
however, they are extremely unstable (Zhu, Zhang, Tsang,
Huang, & Chen, 1997). The study conducted by Zhu et al.
(1997) demonstrated that ()-epigallocatechin gallate and
()-epigallocatechin were more unstable than ()-epicatechin
and ()-epicatechin gallate in a basic solution, giving a plau-
sible explanation to the fact that partial absorption of green
tea catechins in red blood cells of mice is attributed to the
instability of ()-epigallocatechin gallate and ()-epigallocate-
chin in the intestine where the pH is neutral or alkaline. In a
high temperature environment, green tea catechins are not
very stable. Heating may cause the conversion of green tea
catechins to their corresponding isomers, a process known
as epimerization. For example, as epimerization can occur
at high temperature, epigallocatechin gallate in green tea ex-
tract may convert to its epimer component gallocatechin gal-
late. Heat treatments decrease the antioxidant activity of
green tea catechins due to oxidation, thermal degradation,
epimerization and polymerization (Ananingsih, Sharma, &
Zhou, 2013).
3. Flavour constituents
The taste and flavour of tea is governed by key chemical com-
ponents which are polyphenols, caffeine, organic acids and
volatile terpenes (Borse, Rao, Nagalakshmi, & Krishnamurthy,
2002). The characteristic taste of green tea extract is made up
of a mixture of bitterness, astringency, meaty (‘‘umami’’) taste,
sweetness and slight sourness. The compounds, which con-
tribute to, characteristic tea taste, are polyphenolic com-
pounds, amino acids and caffeine (Yamanishi, 1995). Volatile
compounds such as terpenoids, alcohols and carbonyl com-
pounds contribute to the aroma of tea. Volatile fractions of var-
ious green teas reported having numerous active aroma
compounds which are responsible for nutty, floral, fruity,
4 JOURNAL OF FUNCTIONAL FOODS
meaty, popcorn-like, metallic, potato, green, cucumber-like,
and hay-like characteristics (Kumazawa & Masuda, 2002). In
a study conducted by Pripdeevech and Machan (2011), the vol-
atile flavour components of different teas growing in Thailand
were evaluated. These authors reported that the utmost vola-
tile flavour components in green tea were found to be hotrie-
nol, geraniol and linalool. In addition, green Assam tea was
reported to contain linalool, geraniol and a-terpineol as the
key flavour constituents (Pripdeevech & Machan, 2011).
In a classic study to understand the taste characteristics of
different types of catechins, Nakagawa (1970) found that (+)-
catechin, ()-epicatechin, and ()-epigallocatechin were bit-
ter with a sweet aftertaste while ()-epicatechin gallate and
()-epigallocatechin gallate were bitter and astringent. In a
follow up study to determine the chemical components that
affect the taste attributes of green tea in Japan, Nakagawa
(1975) discovered that teas containing higher content of cate-
chins and amino acids scored much higher in taste tests by
consumers. His results indicated that the astringency and bit-
terness of green tea was primarily determined by the content
of catechins and other phenolic compounds. The taste of oo-
long tea appears to be unique. According to Chaturvedula and
Prakash (2011), the mellowness and sweetness of oolong tea
are attributed to non-oxidized catechins, thearubigins, some
secondary polyphenolic compounds, caffeine, free amino
acids and related sugars. The astringency of oolong tea is low-
er, and the sweet taste is stronger than those of green tea. The
distinctive taste of black tea is due to the combination of sev-
eral attributes such as bitterness, astringency, sweetness,
malty taste, green/grassy taste, ‘caramel-like’ and ‘hay-like’
characteristics (Alasalvar et al., 2012). Yang and Landau
(2000) reported that the theaflavins are crucial to the charac-
teristic color and taste of black tea. The theaflavins contain
numerous compounds including, theaflavin, theaflavin-3-gal-
late, theaflavin-3 0 -gallate and theaflavin-3,3 0 -digallate (Chan
et al., 2011). The key astringent taste compounds in black
tea are catechins, theaflavins, and flavonol glycosides (Schar-
bert, Holzmann, & Hofmann, 2004). Although compounds in
all three categories are detected as astringent, flavonol glyco-
sides are the most abundant contributors of astringency in
black tea (Scharbert et al., 2004).
Amino acids are the components in green tea that contrib-
ute full-bodied umami flavour and sweetness. Of these amino
acids, more than 50% are theanine, which is unique to tea
(Chaturvedula & Prakash, 2011). The amino acid theanine
has a chemical structure somewhat similar to that of gluta-
mine, with its distinct attribute being a refined, rich flavour
and sweetness. Amino acids other than theanine present in
tea leaves are glutamine, alanine, asparagine, arginine and
serine. The umami taste of green tea is shown to be due to
theanine and serine. It is believed that 70% of the umami
taste in Japanese green tea is due to theanine (Kaneko,
Kumazawa, Masuda, Henze, & Hofmann, 2006). By conducting
molecular and sensory studies, Kaneko et al. (2006) concluded
that theanine, gallic acid, theogallin and succinic acid are the
main compounds responsible for umami taste of green tea.
Besides catechins and caffeine, some amino acids, in particu-
lar, arginine and alanine also contribute to the bitterness of
green tea (Chaturvedula & Prakash, 2011).
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view, Journal of Functional Foods (2013), http://dx.doi.org/10.1016/j.jff.2013.08.011
x x x ( 2 0 1 3 ) x x x –x x x
4. Antioxidant mechanism
Lipid oxidation and development of rancidity is a major chal-
lenge for food manufacturers, reducing shelf-life and altering
the quality and nutritional value of their products. Autoxida-
tion, the most common process leading to oxidative deterio-
ration of food lipids, is a free radical chain reaction that
proceeds through three distinct stages of initiation, propaga-
tion and termination. The generation of primary free radicals
is facilitated by the presence of oxidation initiators such as
light, heat, ionizing radiation, transition metals, metallopro-
teins, oxidants, various homolysis-prone substances and en-
zymes. Lipid hydroperoxides have been identified as
primary products of autoxidation. Typically, lipid hydroperox-
ides are tasteless and odorless. The decomposition of hydro-
peroxides yields aldehydes, ketones, alcohols, hydrocarbons
and acids, which are known as secondary oxidation products
of lipids. In many cases, these compounds are responsible for
off-flavours and off-odors in food. Antioxidants are of interest
to the food industry because they can delay development of
oxidative rancidity in food. An antioxidant is a substance that
retards lipid oxidation either by inhibiting initial free radical
formation or by preventing them from producing more free
radicals which can disseminate the oxidation reaction. These
substances help preserve foods by delaying development of
rancidity, deterioration and discoloration due to lipid oxida-
tion. There are two main categories of antioxidants in relation
to their mechanism of action: primary antioxidants and sec-
ondary or preventive antioxidants. Primary antioxidants dis-
rupt the oxidative free radical chain reaction by donating
electrons or hydrogen atoms from the phenolic hydroxyl
groups and, therefore, stabilize lipid free radicals, as a result,
inhibit or slow down the initiation phase and disrupt the
propagation stage of autoxidation. Secondary antioxidants
deactivate singlet oxygen, chelate metal ions (i.e., iron, cop-
per), absorb ultraviolet radiation, scavenge oxygen and help
regenerate primary antioxidants. For better effectiveness, pri-
mary antioxidants are often used in combination with sec-
ondary antioxidants.
Tea polyphenols, mainly flavonoids, are well-known for
their antioxidant properties. The antioxidant activity of green
tea polyphenols is primarily attributed to the combination of
aromatic rings and hydroxyl groups that assemble their
chemical structure and consequently binding and neutraliza-
tion of lipid free radicals by these hydroxyl groups. Numerous
studies have demonstrated that polyphenols and tea cate-
chins are exceptional electron donors and effective scaveng-
ers of physiologically relevant reactive oxygen species
in vitro, including superoxide anions (Guo et al., 1999; Micha-
lak, 2006; Nakagawa & Yokozawa, 2002; Nanjo et al., 1993;
Velayutham et al., 2008), peroxyl radicals, and singlet oxygen
(Guo et al., 1999; Michalak, 2006). Catechins also exhibit anti-
oxidant activity via chelating redox active transition-metal
ions. Polyphenolic compounds, possess hydroxyl and car-
boxyl groups, are able to bind particularly iron and copper
(Michalak, 2006). There is another mechanism underlying
the antioxidant ability of plant polyphenols, including tea cat-
echins. Transition metal ions typically have the ability to ini-
tiate free radical chain oxidations by decomposing lipid
 JOURNAL OF FUNCTIONAL FOODS
hydroperoxides (LOOH) by the hemolytic cleavage of the O–O
bond and producing lipid alkoxyl radicals. Phenolic antioxi-
dants, including tea catechins, inhibit lipid peroxidation by
binding these lipid alkoxyl radicals. This activity depends on
the structure of the molecules, and the number and position
of the hydroxyl group in the molecules (Milić, Djilas, & Cana-
danovic-Brunet, 1998). Green tea catechins also exhibit anti-
oxidant activity through inhibiting pro-oxidant enzymes
and inducing antioxidant enzymes (Velayutham et al., 2008).
The antioxidant activity of these catechins and the deriv-
atives showed a marked difference depending on the sub-
strate used for evaluation (Wanasundara & Shahidi, 2005).
Green tea catechins, which performed like other hydrophilic
antioxidants such as Trolox (a water-soluble analog of a-
tocopherol) and ascorbic acid, have been shown to be active
antioxidants in bulk oils, but were prooxidants in the corre-
sponding oil-in-water emulsions (Frankel, Huang, & Aesc-
bach, 1997; Frankel, Huang, Kanner, & German, 1994). In
corn oil triacylglycerols system that was oxidized at 50 °C
the antioxidant activities of epigallocatechin, epigallocate-
chin gallate and epicatechin gallate were superior to those
of epicatechin or catechin (Huang & Frankel, 1997). These cat-
echins have also been highly successful in delaying oxidation
of polyunsaturated fatty acids-rich marine and vegetable oils
(Wanasundara & Shahidi, 1998). In the lecithin-containing lip-
osomes system, epigallocatechin gallate was superior to other
compounds such as epicatechin, epigallocatechin, epicate-
chin gallate, and catechin (Huang & Frankel, 1997). In the
oil-in-water emulsions, however, all catechin compounds
demonstrated pro-oxidant activity (Huang & Frankel, 1997).
The improved antioxidant activity observed for tea catechins
in liposomes compared to emulsions has been explained by
the greater affinity of the polar catechins toward the polar
surface of the lecithin bilayers, thus affording better protec-
tion (Huang & Frankel, 1997). In a study conducted by Zhong
and Shahidi (2011), epigallocatechin gallate was structurally
modified to improve its lipophilicity via esterification of epi-
gallocatechin gallate with selected fatty acids, such as stearic
acid, eicosapentaenoic acid (EPA) and docosahexaenoic acid
(DHA). The lipophilized derivatives so produced exhibited
greater antioxidant activity than the original epigallocatechin
gallate molecule.
Several investigators have speculated the mechanism of
antioxidant action of (+)-catechin using some oxidation mod-
el studies (Hirose, Yamamoto, & Nakayama, 1990; Koketsu,
1997; Zhu et al., 2000). As indicated by the proposed mecha-
nism, (+)-catechin molecule can scavenge four lipid free rad-
icals per molecule (Hirose et al., 1990; Koketsu, 1997). The
antioxidant activity of individual tea polyphenols in different
model systems showed a proportional relationship to the
number of hydrogen radical donors of catechins. A synergistic
effect has been observed between green tea catechins and
ascorbic acid and a-tocopherols (Murakami, Yamaguchi,
Takamura, & Matoba, 2003). Catechin, a monomeric flavanol,
is reported to have hydroxyl, peroxyl, superoxide and 1,1-di-
phenyl-2-picrylhydrazyl (DPPH) radical scavenging activities
(Bors & Michel, 1999; Fukumoto & Mazza, 2000). Moreover,
tea catechins have the ability to chelate iron in food model
systems (Tang, Kerry, Sheehan, & Buckley, 2002). Nakao, Taki-
o, and Ono (1998) reported that the peroxyl radical scavenging
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activities of catechin, epicatechin and epicatechin gallate
were ten times higher than those of L-ascorbate and beta-car-
otene. In another study, Nanjo et al. (1996) documented that
DPPH radical scavenging activities of catechin and epicate-
chin are less than epigallocatechin, epicatechin gallate, and
epigallocatechin gallate. As expected, epicatechin is another
monomeric flavanol (Yilmaz, 2006) came across in green tea.
Reports have shown that epicatechin is capable of scavenging
hydroxyl radicals, peroxyl radicals, superoxide radicals, and
DPPH radicals (Bors & Michel, 1999; Fukumoto & Mazza,
2000; Liu, Ma, Zhou, Yang, & Liu, 2000; Yilmaz, 2006). Yashin,
Yashin, and Nemzer (2011) examined the antioxidant activity
of different types of teas that include green, oolong, black and
pu-erh teas. The antioxidant activity of teas descends in the
following order: green > oolong > black > pu’erh tea. Chan
et al. (2011) evaluated the role of non-polymeric phenolic
(NP) and polymeric tannin (PT) constituents in the antioxi-
dant and antibacterial properties of six brands of green, black,
and herbal teas. Extraction yields of the products ranged from
12% to 23%. Much higher fractionation yields more NP con-
stituents (70–81%) than PT constituents (1–11%), which sug-
gested that, the former are the principal tea components. In
general, the antioxidant properties of green tea extracts were
stronger than those of black and herbal teas (Chan et al.,
2011). For all six brands tested, antioxidant properties of PT
fractions were significantly higher than crude extracts and
NP fractions. Although PT constituents have stronger antiox-
idant and antibacterial properties, they constitute only a min-
or component of the teas (Chan et al., 2011).
Green tea polyphenols have been shown to be effective
against beta-carotene oxidation. For example, tea catechins
have been able to demonstrate an anti-discoloring effect on
beverages and margarine containing beta-carotene (Koketsu,
1997; Unten, Koketsu, & Kim, 1997). It was suggested that
the discoloration of beta-carotene and the oxidation of unsat-
urated fatty acids progressed by the same mechanism. In par-
ticular, carbon–carbon double bonds in beta-carotene and
unsaturated fatty acids are attacked by radicals, becoming
hydroperoxides. Green tea catechins have the ability to sup-
press the degradation of double bonds as radical scavengers.
It was also suggested that the hydroxyl group at the 5 0 -posi-
tion of the B-ring of the catechin structure mostly contributed
to the anti-discoloring effect. Hence, as described above, it
was speculated that green tea polyphenols delayed the degra-
dation of beta-carotene by acting as antioxidants following
the same mechanism. Among the individual green tea poly-
phenols examined, gallocatechin gallate, epigallocatechin
gallate, epigallocatechin, and gallocatechin showed strong
anti-discoloring effect while epicatechin and catechin
showed almost no activity, and gallic acid had moderate
activity (Unten et al., 1997).
5. Regulatory status
In the United States, green tea extract may be used as flavour
agent with antioxidative properties in various fats, oils and
foods containing fats and oils. Green tea has a history of safe
use as food flavouring and is known for its antioxidative func-
tionality as a secondary effect. Green tea extract is well
known in Asian food. According to the United States Code
6 JOURNAL OF FUNCTIONAL FOODS
of Federal Regulations (21 CFR 101.22, and 21 CFR 182.10), the
term ‘natural flavour’ or ‘natural flavouring’ means the essen-
tial oil, oleoresin, essence or extractive, protein hydrolysate,
distillate, or any product of roasting, heating or enzymolysis,
which contains the flavouring constituents derived from a
spice, fruit or fruit juice, vegetable or vegetable juice, edible
yeast, herb, bark, bud, root, leaf or similar plant material,
meat, seafood, poultry, eggs, dairy products or fermentation
products thereof, whose significant function in food is fla-
vouring rather than nutritional (U.S. Food and Drug Adminis-
tration, 2012). Natural flavours include the natural essence or
extractives obtained from plants listed in sections CFR 182.10,
182.20, 182.40, and 182.50 and part 184 of this chapter, and the
substances listed in section 172.510 of this chapter (U.S. Food
and Drug Administration, 2012). Thus, in the United States,
green tea extract may be labeled as natural flavouring. In
European Union (EU), the definition of natural flavouring sub-
stances, as stated in Council Directive 88/388/EEC, applies to
green tea extracts when used as flavourings, and they can
be used in all food applications, unless regulations restrict
the use of flavourings in specific foods (Official Journal of
the European Union, 2008). Certain green tea extracts are also
regarded as traditional foods in the EU, however, market
clearance of green tea extracts depends on the intended use
as well as the product profile. If the primary function high-
lighted is the antioxidant properties, the green tea extract is
then regulated as an antioxidant and must be specifically ap-
proved. Results of various investigations have confirmed that
green tea polyphenolic compounds are largely responsible for
distinctive aroma, color and taste (Borse et al., 2002; Chat-
urvedula & Prakash, 2011; Pripdeevech & Machan, 2011;
Yamanishi, 1995). Hence, various compounds of green tea ex-
tract that contribute to the antioxidant effects also contribute
to flavour properties. The combination of these functional-
ities may allow innovative solutions such as designing spe-
cific green tea extract products to target certain food
applications. As green tea extract contributes to the flavour
profile of food matrix and helps retard lipid oxidation, it can
be successfully used in a wide array of oils, fats, and foods
containing fats and oils.
6. Method of incorporation into food
Proper incorporation of green tea extract into food products is
imperative to ensure that antioxidant components are thor-
oughly dissolved and/or homogeneously dispersed in the
food matrix. As only a small amount of green tea extract
may be required for adequate shelf-life extension in food,
the method of application may determine the achievement
of the antioxidant effectiveness. The selection of application
methods may depend on the type of food products, process-
ing methods, and the availability of the equipment used for
food processing. Commercial extracts of green tea are mainly
available in a fine-grained powder form. Green tea extract can
be solubilized in water prior to addition into foods. Water-sol-
uble green tea extract has a low viscosity for easy spraying
and homogeneous distribution. Moreover, remarkably little
energy is required to produce the dispersion in water. On
the other hand, green tea extract in powder form is often pre-
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x x x ( 2 0 1 3 ) x x x –x x x
ferred in certain formulations containing dry ingredients as
this enables easier blending. The powder carrier aids the
incorporation and homogeneous distribution of the green
tea extract. Where dry ingredients are part of the formulation,
a powdered green tea extract is often preferred due to easier
blending. Conversely, Green tea extract can be dissolved in a
food-grade solvent to produce an oil soluble or dispersible li-
quid product. Liquid forms of green tea extracts may be added
to food products by direct addition into oils and fats. The fat
or oil may be heated to an elevated temperature (i.e., 40–
60 °C) while stirring. Liquid green tea extract should then be
added slowly into melted fat or oil. After addition is com-
pleted, agitation should be continued for an additional period
of time for complete and uniform distribution of green tea ex-
tract in fats and oils.
7. Application in food products
There is growing consumer interest in the green tea extract-
supplemented products in recent years. Green tea extract
has been used in a variety of food products including bread
(Wang & Zhou, 2004), biscuits, dehydrated apple (Lavelli, Van-
taggi, Corey, & Kerr, 2010) and various meat products (Mit-
sumoto, O’Grady, Kerry, & Buckley, 2005; O’Sullivan, Lynch,
Lynch, Buckley, & Kerry, 2004; Tang, Kerry, Sheehan, & Buck-
ley, 2001). The main application areas of green tea extract
are summarized in Table 1. In internal tests with fat spreads,
green tea extract showed comparable antioxidant perfor-
mance to conventional synthetic antioxidant tert-butylhydro-
quinone (TBHQ) and may be more cost-effective than other
natural sources of antioxidants. This makes it ideal for both
full-fat margarines as well as oxidation-sensitive products
that are low in fat, free of trans fatty acids or have a high con-
tent of polyunsaturated fatty acids. In this study, samples of
40% low-fat spread were produced with green tea extract con-
taining 60 ppm total catechins or 100 ppm TBHQ and stored at
5 °C for 13 weeks. A control sample contained no antioxidant
solution. In an accelerated shelf-life test, a comparison of
green tea extract and TBHQ revealed similar performance.
Lipid oxidation is one of the primary causes of quality
deterioration in meat and poultry products. Red meats and
poultry are typically highly susceptible to lipid oxidation
due to their high fat content (Yilmaz, 2006) and a high propor-
tion of polyunsaturated fatty acids. These highly unsaturated
phospholipids present in meat and poultry products are
responsible for the development of off-flavours and off-odors
after cooking and subsequent refrigerated storage. This off-
flavour development, also referred to as warmed-over-flavour
(WOF), frequently causes consumers to reject freshly cooked
meat and poultry products. Therefore, these products should
be protected by natural sources of antioxidants such as green
tea extract to protect against WOF.
In a study conducted by DuPont Nutrition & Health labora-
tories, both control and green tea extract-stabilized turkey
burgers were made using standard industry practice. Green
tea extract was added, at a dosage level of 50–100 ppm. Sam-
ples were evaluated for thiobarbituric acid reactive sub-
stances (TBARS) formation, reported as malondialdehyde (a
secondary product of lipid oxidation) equivalents. Results
 JOURNAL OF FUNCTIONAL FOODS x x x ( 2 0 1 3 ) x x x –x x x 7

Table 1 – Prospective applications of green tea extract.

Application Specific oxidation issues

Raw meats Oxidation of red meat pigments, resulting in

an undesirable brown color
Cooked meat, poultry and Susceptible to oxidation, resulting in a
seafood products warmed-over flavor, discoloration and protein degeneration
Ready-to-eat meals Reheating of meat promotes the oxidation process
Cereals, bakery products, confectioneries, Products are susceptible to oxidation due to long shelf-life requirements
snack foods, nuts and nut products
Oil-in-water emulsions Large oil–water interface and complex food matrix increase susceptibility
(mayonnaise, salad dressings, soups and sauces) to lipid oxidation
Water-in-oil emulsions Large water–oil interface and complex food matrix increase susceptibility
(margarine and fat spreads) to lipid oxidation
Vegetable oils, marine oils, Low oxidative stability, due to trans fatty acid regulations, increases the need for
frying oils, and shortenings enhanced antioxidant protection
Beverages (carbonated and non-carbonated beverages, Products are susceptible to oxidation due to long shelf-life
energy drinks, soft drinks, and juices) requirements

Fig. 2 – TBARS of roasted turkey burgers during storage at Fig. 3 – Effect of green tea extract and BHA on the inhibition
4 °C. of TBARS of roasted turkey burgers during storage at 4 °C.

showed that oxidation in roasted turkey burgers was signifi-

cantly retarded using green tea extract, at levels of 50 to
100 ppm. Samples supplemented with green tea extract had
lower levels of TBARS during storage as compared to control
with nothing added (Fig. 2). In another study conducted by
DuPont Nutrition & Health, the efficacy of green tea extract
was compared with equivalent dosages of butylated hydroxy-
anisole (BHA) and revealed that a dosage of 200 ppm of green
tea extract simply provided the same antioxidant protection
as 40 ppm BHA in chilled (4 °C) roasted turkey burgers during
storage (Fig. 3). The sensory evaluation of the products further
confirmed the advantage of using green tea extract compared
to BHA. In a different study conducted by DuPont Nutrition &
Fig. 4 – Hexanal contents of roasted beef burgers during
Health, the addition of 150 ppm green tea extract was ade-
storage at 4 °C.
quate to inhibit hexanal development (a secondary product
of lipid oxidation) during 13 days of storage in roasted beef
burgers (Fig. 4). This study compared the antioxidant capacity The inhibitory effect of green tea extract on the oxidation
obtained from 150 ppm green tea extract with that obtained of meat lipids was also shown by other investigators. Studies
from 375 ppm rosemary extract. Green tea and rosemary conducted by Tang et al. (2001) and Mitsumoto, O’Grady,
extract-supplemented samples had low levels of hexanal Kerry, and Buckley (2005) showed a high antioxidant activity
throughout storage when compared with the control contain- of green tea catechins in beef and chicken meat. In another
ing nothing added. Furthermore, green tea extract-supple- study, O’Sullivan, Lynch, Lynch, Buckley, and Kerry (2004)
mented sample had lower levels of hexanal as compared to evaluated the effect of several natural extracts on oxidative
the rosemary extract-treated sample throughout storage stability of chicken nuggets during refrigerated (4 °C) storage.
(Fig. 4). Nuggets were supplemented with rosemary extract (at

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8 JOURNAL OF FUNCTIONAL FOODS
Fig. 5 – Color of raw ground beef patties treated with green tea extract and rosemary extract compared to an unprotected
control sample.
1000 ppm), sage extract (at 1000 ppm), or tea catechins (at
100 ppm). Their results indicated that the addition of natural
extracts, including tea catechins, reduced lipid oxidation in
chicken nuggets both in the presence and absence of salt.
However, when sodium tripolyphosphate (STPP) was incorpo-
rated into the same product system, the efficacy of natural ex-
tracts was significantly reduced.
In addition to providing antioxidant activity and natural
flavor attributes, green tea extract can also help improve the
color stability of fresh meat products. Fresh meat color de-
pends on myoglobin that stores oxygen for aerobic metabo-
lism in the muscle. Iron is a pivotal player in meat color.
One of the defining factors of meat color is the chemical state
of iron. Oxymyoglobin gives meats the red color. The brown-
ish color is due to metmyoglobin formed by oxidation of oxy-
myoglobin. However, how green tea extract influences the
color cycle in meat products is not well understood. Studies
conducted by DuPont Nutrition & Health have shown that
the red color of raw ground beef patties (20% fat) was signifi-
cantly improved by the addition of rosemary extract
(1000 ppm) and green tea extract (250 ppm) compared to the
development of browning in control samples (Fig. 5). All sam-
ples were stored at 0–2 °C for 22 days and packaged under
modified atmosphere (80% oxygen and 20% carbon dioxide)
packaging conditions. Determination of redness of the meat
(‘a’ value using Minolta Colorimeter) corresponded with the
development as the visible red color. The meat had lost
almost if not all visible red color when the ‘a’ value reached
approximately 12. The control reached this point around
day 9, and the treated samples around day 16. This shows
that 1000 ppm rosemary extract and 250 ppm green tea
extract exhibited good protection of color for several days be-
yond the control sample, and there was a minimal difference
between the two treatments (Fig. 5). Sensory evaluation of the
burgers showed that green tea extract had no unfavorable fla-
vour impact on the finished burgers where slight rosemary
flavour could be detected in the samples containing rosemary
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extract. In addition, green tea extract contributed to ‘‘meaty’’
(umami) taste in finished burgers (data not shown). In flavour
sensitive meat systems, green tea extract would be a more
viable option than the rosemary extract due to similar effec-
tiveness, but lower flavour impact. A study by Moawad, Abo-
zeid, and Nadir (2012) has shown that the use of tea catechins
in combination with nitrite was more effective in keeping
quality of dry-cured sausages during refrigerated storage as
indicated by attractive red color (a* value) as compared to
samples treated with nitrite. In another study conducted by
DuPont Nutrition & Health laboratories, cured ham slices con-
taining 500 ppm sodium ascorbate and 300 ppm green tea ex-
tract enhanced color stability (from 14 days to 25 days) by
maintaining the redness remarkably efficiently as compared
to a standard ham with 500 ppm sodium ascorbate. The
ham slices were packaged under MAP conditions (80% oxygen
and 20% carbon dioxide) and stored at 5 °C for a period of
38 days. This was shown visually and measured by a Videom-
eterLab 2 – High Performance Multispectral Imaging by
recording a-values. The a-value of 17 was defined as the work-
ing limit for redness acceptability of cured ham.
Results of numerous investigations have also confirmed the
antioxidant properties of green tea catechins in various lipid
and food model systems (Amarowicz & Shahidi, 1995; Chen,
Chan, Ma, Fung, & Wang, 1996; Frankel et al., 1997; Huang &
Frankel, 1997; Koketsu & Satoh, 1997; Matsuzaki & Hara,
1985; Mildner-Szkudlarz, Zawirska-Wojtasiak, Obuchowski, &
Goslinski, 2009; Roedig-Penman & Gordon, 1997; Wanasundara
& Shahidi, 1998). When green tea catechins were added to noo-
dles and to the edible oils (soybean, fish, lard) they were able to
improve the oxidative stability of the fried product and the oil
used for frying (Frankel et al., 1997; Koketsu & Satoh, 1997). In a
study with marine oils such as seal blubber oil and menhaden
oil, both antioxidant and pro-oxidant effects of green tea ex-
tracts have been reported (Wanasundara & Shahidi, 1998).
Green tea extracts demonstrated a pro-oxidant effect in both
oils evaluated, possibly due to the presence of their chlorophyll
 JOURNAL OF FUNCTIONAL FOODS
components. Consequently, in follow-up studies, a column
chromatography was utilized to eliminate chlorophyll com-
pounds from green tea extracts. The dechlorophyllized green
tea extract so produced was incorporated into both seal blub-
ber and menhaden oils at various levels. The antioxidant activ-
ity of dechlorophyllized green tea extract was compared with
that of the butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), TBHQ and a-tocopherol. At the levels
of P200 ppm, dechlorophyllized green tea extract showed
exceptional antioxidant activity in both oils examined. More-
over, the efficacy of dechlorophyllized green tea extract was
superior to that of the BHA (at 200 ppm), BHT (at 200 ppm)
and a-tocopherol (at 500 ppm), but inferior to that of TBHQ
(at 200 ppm). In several other studies, antioxidant activity of
tea polyphenols in vegetable oils and animal fats was exam-
ined (Chen et al., 1996; Koketsu, 1997; Koketsu & Satoh,
1997). Tea polyphenols evidently demonstrated an inhibitory
effect on oxidation of lard, and this effect was dose-depen-
dent. Furthermore, there was a distinct suppressive effect of
tea polyphenols on oxidative rancidity of soybean oil (Koketsu,
1997). When green tea extract was compared with BHA and
tocopherols, green tea catechins showed better antioxidant
activity in fish oil (Koketsu & Satoh, 1997). The ethanol extracts
of green tea catechins strongly inhibited the oxidation of cano-
la oil at 100 °C as shown by measurement of oxygen consump-
tion, whereas extracts of black tea showed little to no
antioxidant activity (Chen et al., 1996). Crude green tea extract
powders were more effective than a-tocopherol and BHA in
lard under conditions of the active oxygen method (AOM) at
97.8 °C (Matsuzaki & Hara, 1985). In a b-carotene-linoleate
model system, ()-epicatechin-3-gallate exhibited the stron-
gest antioxidative activity whereas ()-epigallocatechin had
the weakest performance (Amarowicz & Shahidi, 1995). Fur-
thermore, the antioxidative efficacy of ()-epicatechin and
()-epigallocatechin-3-gallate was similar and in between
those of ()-epicatechin-3-gallate and ()-epigallocatechin.
In another study, Mildner-Szkudlarz et al. (2009) examined
the feasibility of using green tea extract to improve the oxida-
tive stability of biscuits during storage. The antioxidant activ-
ity of green tea extract was compared with commonly used
synthetic antioxidant BHA. Phenolic compounds of green tea
extract had significantly higher DPPH radical scavenging activ-
ity than BHA. However, both green tea extract and BHA were
effective inhibitors of the decomposition of hydroperoxides.
Moreover, biscuits treated with green tea extract had better
sensory profile during storage as compared to samples con-
taining BHA.
8. Stability tests
Oxidative stability is a decisive factor in the processing and
marketing of oils, fats, and lipid-bearing foods. Methods for
determining oxidative stability are, therefore, essential partic-
ularly when green tea extract is being evaluated for its effec-
tiveness in delaying lipid oxidation in these products.
Consequently, many protocols have been developed in at-
tempts to assess the effectiveness of green tea extract in food.
Oxidative stability of green tea extract-supplemented food
products may be determined by storing samples at normal
use conditions and examining them periodically for organo-
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leptic (odor and flavour) changes or by testing them chemically
for oxidative rancidity. Although normal use conditions are
ideal, this protocol may be too slow to be of practical value.
Current methods for determining the effectiveness of green
tea extract against lipid oxidation in food are discussed in this
section.
8.1. Peroxide value (PV)
The peroxide value (PV), an indicator of primary products of
lipid oxidation, has been the most commonly employed
chemical assay for evaluating oxidative stability of fats and
oils. In this test, the melted fat or oil sample is titrated with
a mixture of glacial acetic acid–chloroform (3:2, v/v) and sat-
urated potassium iodide (KI) solution. As hydroperoxides
present in the sample oxidize iodide to iodine, the liberated
iodine is then titrated with standardized solution of sodium
thiosulfate to an end-point. This method is highly empirical,
and any discrepancy in the test procedure may result in a var-
iation of results. The PV is calculated by determining all sub-
stances which oxidize KI, in terms of milli-equivalents of
peroxides per kilogram of sample (Shahidi & Zhong, 2005).
These substances are generally assumed to be peroxides or
other similar products of oil oxidation. The content of hydro-
peroxides can be calculated directly by titration of iodine so
produced. In order to avoid the use of chloroform, the Amer-
ican Oil Chemists Society (AOCS, 2003) has developed an
alternative method, which uses isooctane as solvent (AOCS,
2003; Official Method Cd 8b-90), although the method is lim-
ited to PV of less than 70 meq/kg as described in the AOCS
guidelines. The results of this test are linked to flavor precur-
sors such as hydroperoxides, but not to the actual flavour
compounds. In an internal study, samples of 40% low-fat
spreads were produced with green tea extract containing
60 ppm total catechins or 100 ppm TBHQ and stored at 5 °C
for 13 weeks. A control sample contained no antioxidant solu-
tion. While the hydroperoxides content has significantly in-
creased in the control sample, the samples containing green
tea extract or TBHQ had significantly fewer hydroperoxides
after 15 weeks of storage. In another study, Mustafa (2013)
demonstrated that green tea extract inhibited lipid hydroper-
oxides formation in ground beef during refrigerated storage
(Mustafa, 2013).
8.2. Conjugated dienes
Oxidation of highly unsaturated fatty acids is associated with
an increase in the ultra violet (UV) absorption of the product.
During lipid oxidation, double bonds in unsaturated fatty
acids are changed from non-conjugated to conjugated double
bonds. This process takes place almost immediately after
hydroperoxides have been formed (Gunstone & Norris,
1983). These conjugated diene hydroperoxides have a UV
maximum at 230–240 nm, with a strong characteristic absorp-
tion at 232 nm. Formation of conjugated dienes is an indicator
of detection of primary changes in lipid oxidation. Compared
to peroxide value determination, the measurement of conju-
gated dienes is much simpler and faster and does not rely on
chemical reactions (Shahidi & Zhong, 2005) or formation of
color development. The inhibition of formation of conjugated
10 JOURNAL OF FUNCTIONAL FOODS
dienes by green tea extract or other sources of antioxidants
can be used as means of assessing antioxidant performance
in food. Formation of conjugated dienes is reported to corre-
late well with peroxide values during the early stages of lipid
oxidation (Wanasundara, Shahidi, & Jablonski, 1995). During
Internal testing on 40% fat spreads, the addition of 300 ppm
green tea extract was adequate to inhibit conjugated dienes
formation during 13 weeks of storage at 5 °C. In another inter-
nal study, samples of 40% low-fat spreads were prepared with
green tea extracts containing 40 ppm catechins, and 60 ppm
catechins or 100 ppm TBHQ solution and stored at 5 °C for
15 weeks. The addition of green tea extract had a dose-depen-
dent effect on the inhibition of conjugated dienes. Moreover,
the addition of green tea extract with 60 ppm catechins and
100 ppm TBHQ inhibited the development of conjugated
dienes most effectively throughout the storage period.
8.3. Thiobarbituric acid (TBA) value
TBA is one of the most widely used methods of assessing the
extent of lipid oxidation in foods. The TBA value is typically
expresses in milligrams of malondialdehyde (MDA) equiva-
lents per kilogram of sample, as determined by the methods
described. This assay is derived from the color reaction be-
tween TBA reagent and oxidation products of lipids. Although
most food products naturally contain some level of malondi-
aldehyde as a secondary product of lipid oxidation, especially
affected foods are fried foods, cheese, cooked meats and
dehydrated foods. This assay utilizes the reaction of MDA
with TBA to produce a pink MDA–TBA adduct, which has an
absorbance maximum at 530–540 (532) nm and molar extinc-
tion coefficient at 155 000 M1 cm1 (Kinter, 1995). The forma-
tion of this adduct requires high temperature (90–100 °C) and
acidic conditions. The MDA-TBA adduct so formed can be
measured spectrophotometrically. The TBA assay has been
widely applied in numerous research studies. Close examina-
tion of this method, however, began to divulge that a number
of compounds other than MDA (i.e., alkenals, alkadienals,
browning reaction products, proteins, sugar degradation
products) also react with TBA to form chromogen products,
which absorb at approximately 532 nm. Hence, the results
of this test can be misleading due to lack of specificity. Thus,
the term ‘‘thiobarbituric acid reactive substances’’ (TBARS) is
currently used instead of TBA (Shahidi & Zhong, 2005). Never-
theless, this is one of the frequently used protocols to detect
oxidative deterioration in lipid-bearing foods. The inhibitory
effect of green tea extract on the oxidation of meat lipids
was reported by He˛ś, Korczak, and Gramza (2007), who inves-
tigated the effect of this additive on lipid oxidation in minced
pork meat under frozen storage conditions. The authors
determined the content of TBARS in the control sample and
in samples containing 500 ppm tea extract, 500 ppm rose-
mary extract and 200 ppm BHT. After 6-month storage of
samples, the content of TBARS in samples containing rose-
mary and tea extracts were significantly lower than those
found in BHT and control samples. In another experiment
with a meat model system, antioxidant activity of green tea
catechins as evaluated by TBA values was epigallocatechin
gallate  epicatechin gallate > epigallocatechin > epicatechin.
The inhibitory effect on TBA values of catechins was concen-
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x x x ( 2 0 1 3 ) x x x –x x x
tration dependent, being highest at 200 ppm (Shahidi & Alex-
ander, 1998).
8.4. Oil Stability Index (OSI)
The Oil Stability Index (OSI) is an American Oil Chemists’
Society approved method (AOCS, 1997; Official Method Cd
12b-92) that determines the relative resistance of fats and oils
to oxidation. Oxidative stability instrument (Omnion Inc.,
Rockland, MA, USA) or Rancimat (Brinkmann Instruments,
Inc., Westbury, NY, USA) can be employed to determine OSI
of oil samples. Both Rancimat and Oxidative Stability Instru-
ment are commercial pieces of equipment for the automated
determination of oxidative resistance and are well-known by
the fats and oils industry. The same basic principle lies be-
hind both the Rancimat and Oxidative Stability Instrument
(Shahidi & Zhong, 2005). The two instruments differ only
slightly in their design and operating conditions. In this
method, a sample of oil or melted fat is weighed into a glass
test tube. The test tube is then placed in a heating block at a
temperature of typically around 110 °C. However, the OSI may
be run at temperatures of 80–140 °C. A stream of purified air is
passed through the sample, and the effluent stream of air is
bubbled through a collection tube containing deionized water.
An electrode is placed in deionized water, and the conductiv-
ity of the water is continually monitored. As the oil oxidizes,
volatile organic acids are produced and trapped in the collec-
tion tube, which increase the conductivity of the water. This
change in conductivity is monitored by computer software.
The Oil Stability Index (OSI) is defined as the point of maxi-
mum change of the rate of oxidation. In a study conducted
by DuPont Nutrition & Health laboratories, the OSI values of
canola oil containing liquid green tea extract were deter-
mined at 110 °C. The canola oil sample with nothing added
had an OSI value of 19.8 h. When liquid green tea extract
(1300 ppm) was added to the canola oil, the OSI value in-
creased up to 31.4 h. Hence, canola oil with green tea extract
found to be more stable as compared to the control sample
with nothing added.
8.5. Oxipresä
The Oxipresä (Mikrolab Aarhus A/S, Hojbjerg, Denmark), a
method of examining accelerated stability of heterogeneous
food and feed products, is a modification of the traditional
ASTM oxygen bomb method. In this method, the food or feed
samples are placed in a closed container with an oxygen head-
space. Oxidation is accelerated by heating (to 90–140 °C) and by
the use of oxygen under pressure (initial oxygen pressure of 5
bars). As the product oxidizes, the oxygen content of the head-
space gas decreases which results in a decline of pressure
within the closed container. This reduction in pressure is mea-
sured electronically. Induction period can be determined by
plotting the pressure over time. This induction period (IP) is as-
sumed to be a measure of the resistance of the lipids toward
oxidation. For ease of interpretation, the amount of oxygen
consumed by the sample can be calculated and plotted against
time. Products have a large and fast oxygen consumption (as
indicated by short IP) will be more prone to oxidation, and pre-
sumably have a shorter shelf-life. The Oxipresä is normally
 JOURNAL OF FUNCTIONAL FOODS
equipped with three thermostatic aluminum heating blocks,
each with two holes allowing six oxygen bombs to be operated
simultaneously at a maximum of three different tempera-
tures. The apparatus may also be equipped with only two oxy-
gen bombs. It is suitable for testing the expected shelf-life of
products with multi-phase systems and systems containing
water (i.e., margarines, fat spreads, mayonnaise, salad dress-
ings, snacks, peanuts, fish meal, chicken meal, meat products,
etc.), which cannot be tested using the OSI. In a study con-
ducted by DuPont Nutrition & Health laboratories, both control
and green tea extract-stabilized fat spreads were made using
standard industry practice. Green tea extract was added at a
dosage level of 200 ppm. Samples were evaluated for Oxipresä
induction times at 100 °C. Results showed that oxidative stabil-
ity of fat spreads was significantly improved using the green
tea extract. Samples supplemented with green tea extract
had higher induction periods as compared to control with
nothing added.
8.6. Schaal oven storage test
The Schaal oven test, also referred to as oven storage test, is
often used for evaluating oils, fats, and foods containing fats
and oils. The AOCS (1997) official method Cg 5-97 describes
the protocol for an accelerated test to measure the stability
of oil by aging in an oven at 60 °C. Methods such as peroxide
value, conjugated dienes, TBARS, sensory evaluation, gas
chromatographic volatile com pounds are then employed to
measure oxidation levels and endpoints in the oils. Because
this test uses relatively low temperatures, the samples are ex-
posed to mild oxidative stress. The results of the Schaal oven
test normally correlate exceptionally well with actual shelf-
life predictions because the end-point represents a lower de-
gree of oxidation (Frankel, 2005). Under Schaal oven test con-
dition at 65 °C over a period of 144-h, seal blubber and
menhaden oils treated with tea catechins showed excellent
oxidative stability as compared to samples containing BHA,
BHT, TBHQ, and a-tocopherol. The antioxidant capacity of
catechins in prevention of marine oils oxidation was in the
order of epicatechin gallate > epigallocatechin gallate > epi-
gallocatechin > epicatechin. Moreover, the epigallocatechin
was slightly more effective than TBHQ in the system used
(Wanasundara & Shahidi, 1996).
8.7. Sensory evaluation
Sensory evaluation is a scientific method that is used to mea-
sure, analyze, recall, and interpret the reactions to those char-
acteristics of foods as they are perceived through the human
senses of sight, smell, taste, touch and hearing. Sensory test-
ing requires panels of human participants, on whom the
products are evaluated, and then record the responses made
by them. The sensory quality of a food product is one of the
most crucial aspects influencing its success in the market-
place. All sensory testing protocols are typically categorized
into three broad groups: hedonistic, discriminative and
descriptive testing. Among these, hedonistic tests are always
used within the scope of consumer evaluations and serve to
characterize the consumer behavior. On the other hand, both
discriminative and descriptive sensory tests may only be
Please cite this article in press as: Namal Senanayake, S.P.J., Green tea extract: Chemistry, antioxidant properties and food applications – A re-
view, Journal of Functional Foods (2013), http://dx.doi.org/10.1016/j.jff.2013.08.011
x x x ( 2 0 1 3 ) x x x –x x x 11
carried out by trained/experienced panelists and can give
exceptionally detailed information about individual product
parameters. Hence, the selection of a proper sensory method
is crucial. Sensory evaluation methods are typically expensive
and may require a relatively large number of samples. None-
theless, sensory analyses are typically conducted by the food
industry to evaluate the performance of antioxidants during
shelf-life of food products. Internal studies conducted by Du-
Pont Nutrition & Health have demonstrated that sensory
quality of green tea extract was superior to some commonly
used synthetic antioxidants in chilled (4 °C), roasted turkey
burgers. The WOF in the control sample within 24 h of storage
was unusually high. The WOF of samples treated with green
tea extract was not detected even after 13-days of storage at
4 °C. Moreover, the sensory attributes of green tea extract
were superior to those of the synthetic antioxidant BHA. In
another study, Mildner-Szkudlarz et al. (2009) showed that
the sensory profile of green tea extract-supplemented biscuits
was superior to samples containing BHA.
9. Conclusions
During the past few decades, the use of natural antioxidants
and plant-derived extracts has received increased interest
due to concerns over possible adverse health effects caused
by the use of synthetic antioxidants. Green tea extract, a nat-
ural source of antioxidant, has been successfully used not
only to enhance flavour but also to extend the shelf-life of
various food products. Green tea extract is particularly suit-
able for products with high susceptibility to lipid oxidation,
including trans-free and low-fat products and foods with a
high content of polyunsaturated fatty acids. The primary
application areas of green tea extract include meat, poultry,
seafood, mayonnaise, salad dressings, soups, sauces, marga-
rines, fat spreads, shortening, frying oils, bakery products,
pizza toppings, cereals and snack foods, among many others.
Research reports on measuring the effectiveness of green tea
extract in food against lipid oxidation are scarce in the litera-
ture. The typical parameters measured in determining the
effectiveness of green tea extract in food include peroxide val-
ues, conjugated dienes, thiobarbituric acid reactive sub-
stances (TBARS), Oil Stability Index (OSI), Oxipres induction
period, and sensory evaluation, among others.
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