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Polyphenolic composition and antioxidant activity of Kenyan Tea cultivars


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Polyphenolic fractions in tea are potent bioactive molecules. In this study, the polyphenolic composition of 25 different types of Kenyan tea cultivars was determined using the HPLC and the Folins Ciocalteus spectrophotometric methods. Total Polyphenols, Total Catechins, individual catechins and Antioxidant Activity were significantly (P<0.05) different among tea varieties, with green tea had the highest levels of Total Polyphenols ranging from (19.70-26.12%), TC (8.51%-17.60%), individual catechins, and AA (86.65-94.50%). In vitro bioassay carried out using 2, 2'-diphenyl picrylhydrazyl radical showed epigallocatechin gallate was the most potent catechin and the most potent in antioxidant activity (r=0.968***). Epigallocatechin (r=0.659***, p<0.001), Epicatechigallate (r=0.454*, p<0.001) and Epicatechin (EC) (r=0.780***, p<0.001) showed significant (p<0.05) antioxidant activity. Black tea contained high levels of Theaflavins and Thearubigins (2.072% to 17.12%), respectively) which accounted for its antioxidant activity (r=0.803*** and r=0.859***, respectively). Gallic acid also showed significant (r=0.530*) contribution to the antioxidant activity in black tea. Data obtained from this study reveals that different Kenyan tea cultivars have different polyphenolic composition which imparts on their unique biochemical qualities. Green and white tea products are rich in catechins, black tea products are rich in TFs and TRs while purple teas are rich in anthocyanins.
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Journal of Pharmacognosy and Phytochemistry 2014; 3(4): 105-116
E-ISSN: 2278-4136
P-ISSN: 2349-8196
JPP 2014; 3(4): 105-116
Received: 24-09-2014
Accepted: 09-10-2014
Karori S.M
Department of Biochemistry and
Molecular Biology, Egerton
University. P.O Box 536- 20115,
Egerton, Kenya.
Wachira F.N
Association for Strengthening
Agricultural Research in East and
Central Africa, P.O Box 765,
Entebbe, Uganda.
Ngure R.M
Department of Veterinary Clinical
Studies, Egerton University. P.O
Box 536-20115, Egerton, Kenya.
Mireji P.O
Department of Epidemiology of
Microbial Diseases, Yale School of
Public Health, 607 Laboratory of
Epidemiology and Public Health
,60 College St .New Haven, CT
06510, USA.
Karori S.M
Department of Biochemistry and
Molecular Biology, Egerton
University. P.O Box 536- 20115,
Egerton, Kenya.
Polyphenolic composition and antioxidant activity of
Kenyan Tea cultivars
Karori S.M, Wachira F.N, Ngure R.M and Mireji P.O
Polyphenolic fractions in tea are potent bioactive molecules. In this study, the polyphenolic composition
of 25 different types of Kenyan tea cultivars was determined using the HPLC and the Folins Ciocalteus
spectrophotometric methods. Total Polyphenols, Total Catechins, individual catechins and Antioxidant
Activity were significantly (P<0.05) different among tea varieties, with green tea had the highest levels
of Total Polyphenols ranging from (19.70-26.12%), TC (8.51%-17.60%), individual catechins, and AA
(86.65-94.50%). In vitro bioassay carried out using 2, 2’-diphenyl picrylhydrazyl radical showed
epigallocatechin gallate was the most potent catechin and the most potent in antioxidant activity
(r=0.968***). Epigallocatechin (r=0.659***, p<0.001), Epicatechigallate (r=0.454*, p<0.001) and
Epicatechin (EC) (r=0.780***, p<0.001) showed significant (p<0.05) antioxidant activity. Black tea
contained high levels of Theaflavins and Thearubigins (2.072% to 17.12%), respectively) which
accounted for its antioxidant activity (r=0.803*** and r=0.859***, respectively). Gallic acid also showed
significant (r=0.530*) contribution to the antioxidant activity in black tea. Data obtained from this study
reveals that different Kenyan tea cultivars have different polyphenolic composition which imparts on
their unique biochemical qualities. Green and white tea products are rich in catechins, black tea products
are rich in TFs and TRs while purple teas are rich in anthocyanins.
Keywords: Catechins, EGCG, Theaflavins, Thearubigens, Anthocyanins, Antioxidant Activity.
1. Introduction
Tea, from Camellia sinensis L.O Kuntze is one of the most widely consumed beverages in the
world and it was first introduced in Kenya in1904 by the British settlers. The crop has
expanded to cover an area of around 157,720 ha in the highlands East and West of the Great
Rift Valley in Kenya [1]. The tea plant is an evergreen bush that grows to 15 m high in the wild,
and 60–100 cm under cultivation. Tea in cultivation forms a table from which the young
leaves are harvested through and a cyclic pruning is carried out after every 3 to 4 years and
commercial harvesting is carried out either by hand or machine [2].
Young leaves of tea are processed into different types of products, the predominant ones being
black, green, white and oolong tea. Green tea is mainly consumed in China, Japan and the
Middle East, while black tea is mostly consumed in India, Sri-Lanka, European countries and
regions of Africa. Popularity of tea is due to its aroma, pleasant taste and medicinal benefit [3].
Tea from Kenya tea is better perceived since it is grown free of agrochemicals in an ideal
environment that naturally deters to pests and attack by plant diseases. This pleasant natural
condition guarantees the consumer the safest and most refreshing sought after health drink in
the world. Phytochemicals and functional components in tea are receiving a lot of attention
due to their potential benefits in health when consumed as part of a varied diet on a regular
basis and at effective levels. Many nutraceuticals, functional foods and naturally occurring
polyphenols have physiological and pharmacological activities including their well
characterized antioxidant properties [4, 5]. Since the scientific community and food industry
communities share a common goal of extending the quality of human life, through
development of viable options for the management of chronic diseases through the use of
nutraceuticals, functional foods have become a potential starting point. This is because
functional foods are fairly affordable, readily available and extremely active, have profound
effect on cell metabolism and often demonstrate few side effects [6]. It is evident that
nutraceuticals offer a selective advantage over synthetic drugs necessitating need to investigate
their usefulness to human health.
Journal of Pharmacognosy and Phytochemistry
Despite the use of tea in food and drinks, it has increasingly
been put to many uses. For example, numerous
environmentally friendly industrial cleaning agents,
deodorizers and anti-microbial agents have been formulated
using tea [7, 8]. However, data to support the view that tea is
pharmacologically active has been generated particularly
using green tea, which is widely consumed in Asia [9, 10]. As a
result, green tea has been widely marketed as a health
product since the chemical composition is well characterized.
However, the much information on the chemical composition
of black aerated or fermented tea, the principle type of tea
product consumed in Kenya is based broadly on the
theaflavins and thearubigens but none of the theaflavins
fractions. In addition, information on the chemical
composition of purple tea, a novel product released recently
by the Tea Research Institute (TRI) is lacking. There is need
therefore to conduct systematic research on the
phytochemical composition of Kenyan tea cultivars
especially black tea and purple teas in order to generate
requisite data which can be used to understand the
pharmacological activity of tea.
2.1 Tea Products and Determination of Flavonoid Levels
2.1.1 Tea samples
The tea samples were sourced from Tea Research
Institute (TRI), Timbilil Estate, Kericho (latitude 0˚ 22’S,
longitude 35˚ 21’E, altitude 2180 m above mean sea level
and processed at the TRFK miniature factory as described by
Karori et al., [5].
2.1.2 Biochemical profiling of the tea extracts based on
A modified method of Zuo et al., [11] which is based on high
performance liquid chromatography was used to assay for the
tea catechins of as described by Kerio et al., [12].
2.1.3 Determination of total polyphenols in the tea
The Folin-Ciocalteu phenol reagent method was used to
determine total polyphenols in the tea extracts according to
ISO (BS ISO 14502-1: 2005(E)).
2.1.4 Analysis of the content of total theaflavins and
individual theaflavin fractions content in the tea samples
Black, green, purple and white teas were also assayed for
total theaflavins (TF) using the flavognost method of Hilton
and Palmer Jones [13] whereas individual theaflavins fraction
ratio were determined as described by Kilel et al., [14].
2.1.5 Spectrophotometric determination of total
thearubigins in the tea samples
Total thearubigins (TRs) were determined in the tea samples
using the method of Roberts and Smith [15].
2.1.6 Determination of total monomeric anthocyanin
The total monomeric anthocyanin content in the processed
aerated, unaerated purple tea samples was determined in
duplicate using the pH differential method Kerio et al., [12].
2.1.7 Determination of antioxidant activity of tea
The stable DPPH (2,2-diphenyl-1-picrylhydrazyl) radical
was used for determination of free radical scavenging of tea
extracts using a modified method of Brand-Williams et al.,
2.1.8 Statistical analysis
All statistical analysis was carried out using MSTAT-C
statistical software. ANOVA was used to determine the
means, coefficient of variation and any differences between
the samples. Least Significance Difference (LSD) was used
separate means. The probability limit was set at p0.05
significant level. Results of the parameter determined were
expressed as a mean of the triplicate determination.
3. Results and Discussion
3.1 Chromatographic and Spectrophotometric Analysis
3.1.1 Tea polyphenols
This study compared the total polyphenols levels in tea
samples processed from Kenyan germplasm using the Folins-
Ciocalteus method. The green (unaerated), black (aerated),
white and purple teas samples analyzed differed significantly
in the levels of total polyphenols (p0.05). Kenyan green teas
were rich in total polyphenols with their levels ranging from
the highest amount of 26.1% for cultivar Ejulu-L to the
lowest of 19.7% for cultivar TRFK 430/12 as shown in Table
1. Black teas had a lower total polyphenol concentration
compared to green teas with cultivar Ejulu-L having 21.2%
and cultivar TRFK 301/4 the lowest value of 19.7%. It was
however evident, that some black teas from Kenya had a
higher polyphenolic concentration than green teas. Cultivar
TRFK 6/8, a high quality Kenyan genotype used in this study
as an internal standard for black tea quality, recorded a total
polyphenol content of 25.13% and 20.72% for unaerated and
aerated tea respectively, which was not significantly different
(p0.05) from tea cultivar Ejulu-L. Total polyphenol content
of aerated and anaerated teas processed from purple coloured
leaf cultivars was 20.03% and 21.90% respectively while
white teas processed from plucked shoots of cultivar AHP
S15/10 was 22.43% (Table 1).
Polyphenols which are constituents of secondary metabolism
in plants play a role in plant defense mechanism against
insects, birds and animals. This study revealed that these
chemicals are retained almost intact in unaerated processed
teas. Unaerated Green tea is made without enzymatic auto-
oxidation of polyphenols, since the enzyme polyphenol
oxidase is inactivated by heat during the early stages of
processing [17]. This process ensures that polyphenols present
in green tea are nearly the same as those found in fresh tea
leaves. In a broad sense, green tea polyphenols consist of
simple and complex compounds, the large majority of which
are the flavonoid monomers catechins, catechin gallates and
flavonols [18, 19].
Polyphenols occurring in black tea usually consist of residual
green tea polyphenols such as catechins flavonols and
oxidation products of green tea polyphenols such as
theaflavins and thearubigins [19]. Some catechins and catechin
gallates may be epimerized or degallated during the
processing of black tea. Most of the catechins and their
gallates undergo known enzymatic oxidation to form more
polymeric polyphenols that are characteristic of black tea,
namely theaflavins and thearubigins [19, 20]. Therefore, the
amount of polyphenols in green tea is higher than that of
black teas since the auto-oxidation process results in a
significant conversion of green tea polyphenols to highly
polymerized substances such as theaflavins which contribute
Journal of Pharmacognosy and Phytochemistry
to the characteristic bright orange color of black tea and
thearubigens which are more chemically heterogeneous and
tend to be brownish-red [20, 21]. However, despite this
observation, the exact contribution of inter-flavonoid
linkages and the general structure of thearubigins to the
above quality parameters remains ambiguous and their
structures remain speculative. There is need therefore to
elucidate the thearubigins structure to help in optimizing the
tea processing parameters which might contribute to the
customer’s needs.
Table 1: Total polyphenols (TP) (%) and Total Catechins (TC) (%) levels of different tea products used in this study
Clone Green Tea (TP) Black Tea (TP) Green Tea (TC) Black Tea (TC)
TRFK 301/4 22.64de
14.96h 16.14cde
TRFK 301/5 22.07defg 15.91gh 16.42bcde 3.515hijkl
TRFK 303/216 21.42def 16.41fgh 15.78fgh 2.635l
TRFK 303/231 25.61ab 18.85cde 15.41ghi 3.680hijkl
TRFK 303/577 23.21def 23.21def 16.60bc 7.215ab
TRFK 303/745 20.85ghi 18.35de 11.26
TRFK 337/138 20.85ghi 19.76abcd 17.60a 6.335bc
TRFK 371/3 24.75ab 20.38abc 14.93i 4.25efghi
TRFK 430/3 22.74de 18.22de
14.20j 3.820ghij
TRFK 430/4 19.98hi 17.56efg 9.52n 3.225ijkl
TRFK 430/12 19.70i 16.15gh 11.90l 3.900fghij
TRFK 430/63 21.26fgh 15.98gh 15.31hi 3.700ghijk
TRFK 430/90 23.25cd 19.65abcd 15.89efg 8.115a
TRFK 524/170 21.84defg 17.56efg 11.90l 3.82ghij
TRFK 524/48 22.70de 17.52efg 16.39bcde 3.665hijkl
BBK35 24.55
c 21.03a 16.04de
EPKC12 22.03defg 20.38abc 12.14l 5.215de
EPK D 99/10 21.59efg 17.39efg 14.15j 3.150jkl
EJULU-L 26.12a 21.22a 16.53
AHPS 15/10 22.43def 18.94bcde 15.99ef 5.000de
EPK TN 14/3 21.92defg 17.94efg 10.82
TRFK 6/8 25.13ab 20.72ab 16.85b 6.775bc
TRFK 31/8 21.94defg 20.73ab 14.31j 4.755efg
TRFK 31/11 22.85de 20.68abc 12.77
TRFK 100/5 23.07d 19.09bcde 15.91efg 3.555hijkl
TRFK 306/1
Means within a column followed by the same letter are not significantly different at p0.05 according to DMRT. Purple tea; TRFK 306/1;
White tea; AHPS 15/10
3.1.2 Total catechin content
Results of the total catechins levels in green, black and white
tea products processed from the 25 tea cultivars assayed in
this study are presented in Table 1. The tea cultivars
produced tea products that differed significantly (p0.05) in
total catechin content. Non-aerated (green) teas contained
significantly (p0.05) higher amounts of total catechins of
17.60% to 8.51% than aerated (black) teas which ranged
from 8.115% to 3.150%. These results demonstrated clearly
that the degree of auto-oxidation ‘’fermentation’’ during the
manufacturing process had an influence on the catechin
content of the final product. During this processing, the
polyphenol oxidase enzyme catalyzes the oxidation of
catechins into quinones by molecular oxygen [20, 22]. The
quinones generated from the oxidation of the B-ring in the
dihydroxylated catechins condense with the quinones from
the B-ring of the trihydroxy related catechins to form
theaflavins. Since there is a difference in the reduction
potential, quinones will also take part in the redox
equilibrium at fermentation and this explains the depletion of
catechins at different rates [19, 20]. White tea comes from the
same plant Camellia sinensis as the case of green and black
teas. It is processed from the young buds and/or young leaves
and the descriptive term “white” stems from the high
proportion of silvery buds harvested from the plants to
produce the tea. The buds to manufacture this type of tea are
picked then rapidly steamed and dried, without fermentation,
rolling or roasting. Minimal processing not only protects the
delicate, light and slightly sweet flavor of white tea, but also
enables the retention of high levels of phytochemicals [18, 19].
This explains why white tea had high levels of total catechins
despite being processed only the bud. The composition of tea
leaves varies significantly with shoot maturity and season.
Young leaves are composed of polyphenols in the following
order EGCG > EGC > ECG > EC. As for mature leaves, it is
EGC > EGCG > ECG > EC while old leaves composition
was EGC > EGCG > EC > ECG. It is therefore important to
use young leaves and obey plucking standards to achieve
optimum quality especially for black tea. However, since
mature and old tea leaves possess high amounts of EGC, it
would be advisable to research more on ways they can
utilized for further applications.
Journal of Pharmacognosy and Phytochemistry
Purple leaf coloured cultivar TRFK 306/1 recorded a very
low concentration of total catechins of only 8.51% for
processed green tea. In order to establish why the purple teas
recorded low levels of catechins, an analysis of the total
monomeric anthocyanin content and a fractionation of
anthocyanin by HPLC were carried out. Results from this
experiment revealed that purple tea was instead rich in
anthocyanins which are important phytochemicals found in
the novel purple-pigmented cultivars and not catechins as
earlier thought. Anthocyanins are a sub-class of flavonoids
synthesized via the phenylpropanoid pathway and are wide
spread in the animal kingdom where they present diverse
biological and biochemical interests [23]. The anthocyanidins
fractions in the processed unaerated and aerated tea products
from the purple coloured tea cultivar 306/1 were identified
and quantified by HPLC using pure anthocyanidin
/anthocyanin standards. The order of elution of the
anthocyanidin/anthocyanin was cyanidin-3-O-galactoside <
cyanidin-3-O-glucoside < delphinidin < cyanidin <
pelargonidin < peonidin < malvidin with malvidin being the
most predominant anthocyanin as shown in a representative
HPLC chromatogram of aerated tea from a purple tea cultivar
(Figure 1). The results obtained from this study collaborated
with those of Kerio et al., [12].
Retention time, min
Fig 1: A representative HPLC chromatogram of processed black tea sample from purple colored cultivar TRFK 306/1
KEY: X-axis = Retention time (min) Y-axis = Peak area
However, it was noted that anthocyanidins levels were
significantly higher (p 0.05) in unaerated than in aerated
tea. This can be attributed to the tea processing procedures
where conversion of fresh tea leaf to the aerated tea
decreases the total monomeric anthocyanins. Kerio et al., [12]
observed similar results while characterizing anthocyanins in
Kenyan teas and attributed this to anthocyanin degradation
during the manufacture of black teas. Although no
mechanism has been developed so far on the anthocyanin
degradation, studies in other plants such as strawberry have
been attempted [24]. In their work Liu et al., [24], found out
that anthocyanins were rapidly degraded by PPO in the
presence of other polyphenol compounds such as catechins.
For example, cyaniding-3-O-rutinoside is degraded by PPO
in the presence of (-)-epicatechin and this is responsible for
the formation of dehydroepicatechin A [12, 24]. The coupled
oxidation reactions can be used to explain the sudden
reduction of anthocaynins in the aerated teas. However, this
is not the case in the unaerated teas since PPO is deactivated
by steaming freshly plucked tea leaf and therefore the
formation of the reactive O-quinones is stopped and
subsequently, anthocyanins are not degraded. This is a
hypothesis which needs further studies to establish the exact
cause of degradation.
3.1.3 Catechin fractions
The catechins identified in tea cultivars were EC, EGC, ECG
and EGCG. The two main gallated catechins present were
EGCG and ECG while the others were non-gallated. Beside
the main peaks identified, several minor peaks were also
detected, which indicated that other unidentified catechin
compounds existed in the tea extracts (Figure 2). There was
however great similarity in the HPLC chromatographic
pattern which indicated the close similarity in catechin
profiles in the teas studied.
Catechin fractions assayed in this study were statistically
different (p<0.05) as shown in Table 2. The results obtained
revealed that black (aerated) teas had lower catechin levels
than the green and white (non-aerated) teas (Table 2 and 3).
Individual catechins varied significantly (p<0.05) among the
teas with EGCG, GC and EGC levels being the highest and
+C, ECG and EC being less abundant. These results are
similar to those of Karori et al., [5]. The reduction in the
Journal of Pharmacognosy and Phytochemistry
catechin content in black teas compared to the green tea due
to the monomeric flavan-3-ols undergoing polyphenol
oxidase-dependent polymerization. This results in the
formation of theaflavins, thearubigins, bisflavanols and other
complex oligomers [25, 26, 27].
Retention time, min
Fig 2: A representative high performance liquid chromatogram of green tea cultivar
Table 2: Individual catechin (%) levels of different green tea products analyzed
Individual Catechins
Clone EGCG% EGC% EC% ECG% C%
TRFK 301/4 3.800
2.725j 2.745
4.585ab 2.690a
TRFK 301/5 3.105n 4.000fg 3.280a 4.180c 1.850
TRFK 303/216 4.870hi 5.770
c 1.960cde 2.035ijkl 1.075de
TRFK 303/231 5.555de
1.485fghij 2.465e
TRFK 303/577 5.185fgh 3.695gh 1.775cdefg 7.215ab 1.240c
TRFK 303/745 3.830l
3.370hi 1.355ghij
1.455n 1.250c
TRFK 337/138 5.725cde 4.040fg 1.440fghij
2.275fghi 1.280c
TRFK 371/3 5.525de
4.585e 1.770cdefg 1.990kl 0.9600e
TRFK 430/3 4.990ghi 5.130
1.725cdefg 3.820ghij 0.3200j
TRFK 430/4 3.000n 2.600j 1.275hijkl 1.665mn 1.295c
TRFK 430/12 3.920kl
3.225i 1.460fghij 2.650e 0.6400ghi
TRFK 430/63 5.040ghi 5.790
c 1.695defg 2.275ffghi 0.5100hij
TRFK 430/90 5.305fg 3.215i 1.870cde
TRFK 524/170 3.660
3.470hi 1.600efghi 2.380fg 0.785fg
TRFK 524/48 6.030
c 5.715
c 1.935cde 2.295fgh 0.4200ij
BBK35 5.825c
6.075ab 1.835cde
EPKC12 4.235kl 3.250i 1.555efghij 2.385fg 0.7100fgh
EPK D 99/10 4.305j
1.475fghij 1.992l 0.9150e
EJULU-L 6.625a 5.800
c 2.145c 4.775a 1.310c
AHPS 15/10 5.855c
1.635efghi 2.380fg 0.5800ghij
EPK TN 14/3 3.755
3.560hi 1.225ijkl 1.915l 0.3650j
TRFK 6/8 6.420ab 6.255a 2.120c
c 1.1495c
TRFK 31/8 5.350efg 5.710
c 1.025kl 1.865l
TRFK 31/11 4.885hi 4.005fg 1.135jkl 2.350fg 0.4250ij
TRFK 100/5 4.695ij 5.405c
2.025jkl 0.4900hij
TRFK 306/1
Means within a column followed by the same letter are not significantly different at p0.05 according to DMRT.
Journal of Pharmacognosy and Phytochemistry
White tea which is predominantly manufactured from the young
apical hairy bud only, showed high levels of EGCG and ECG that
are present in higher amounts in fresh young leaves. This result
corroborates the findings of Saijo et al., [28] who determined the
chemical constituents of young tea leaves and the change occurring
during leaf development. The decrease in the gallic acid esters of
catechin such as EGCG and ECG during leaf development means
that there is a slow biosynthesis of gallic acid moiety in each
catechin gallate compared with dry matter production. Since
catechin biosynthesis is slower than dry matter production from
young leaves to the less young leaves, it is apparent that there is no
weight increase in the less young and mature leaves and as a result
catechin moves to other young leaves or are metabolized to other
products. This accounts for the change in catechin levels in various
leaf developmental stages and hence the levels of residual catechins
in tea manufactured from different leaf ages as exemplified in the
differences in catechin levels between white tea and the other types
of teas in this study. Consequently, because of the different rates of
growth among different cultivars, clones will accumulate varying
amounts of catechins in their leaves [4]. Cultivar AHP S15/10, from
which white tea is processed, is a fast growing clone compared to
cultivar TRFK 6/8 from which green and black teas were produced
[29]. The lower total polyphenol content in the white tea than in the
green teas in our assay can be ascribed to the fast growth of cultivar
AHP S15/10 compared to other cultivar such as TRFK 6/8.
Table 3: Individual catechin (%) levels of different black tea products analyzed
Individual Catechins
Clone EGCG% EGC% EC% ECG% C%
TRFK 301/4 0.2950j 0.5050j 0.4250
cdefg 0.8900defgh 0.0650
TRFK 301/5 0.6050gh ij 0.9450hij 0.6050
0.9700defg 0.3900c
TRFK 303/216 0.8600efgh 0.6700ij 0.2600fg 0.6850fghij 0.1650c
TRFK 303/231 0.8100efgh 1.590efghi 0.1750g 0.9650defg 0.2400c
TRFK 303/577 1.330ab 4.065a 0.5050
1.095cde 0.4150c
TRFK 303/745 0.9000defgh 2.530cde 0.2efgh 0.4950ij 0.1150
TRFK 337/138 1.010
cde 1.020de
TRFK 371/3 1.010
1.260ghij 0.3400cdefg 1.100cde 4.25efghi
TRFK 430/3 0.9150cdefgh 1.515fghi 0.4200
cdefg 0.8350efghi 0.1350
TRFK 430/4 0.5550hij 1.655efgh 0.3200defg 0.4250j 0.2500c
TRFK 430/12 0.8450efgh 1.695efgh 0.5800
0.6900fghij 0.2200c
TRFK 430/63 0.7050fghi 1.955efg 0.4350
cdefg 0.4350j 0.3250c
TRFK 430/90 1.295abc
3.805ab 1.300a 1.210
TRFK 524/170 0.3450ij 1.150ghi j 0.5200
0.9850defg 0.7300
TRFK 524/48 0.7250fghi 1.450fghij 0.5100
0.6900fghij 0.8900abc
BBK35 0.5700hij 1.975efg 0.2950efg 0.8750defg 1.045abc
EPKC12 1.010
2.055defg 0.4850
1.105cde 5.215de
EPK D 99/10 0.7950efgh 1.130ghij 0.4850
0.6900fghij 0.7450
EJULU-L 1.315ab 3.190abc 0.5900
c 1.810a 1.545abc
AHPS 15/10 1.000
cdefg 2.065defg 0.2750fg 0.5350hij 1.415abc
EPK TN 14/3 0.7900efgh 1.550fghi 0.3200defg 0.6300ghij 0.9000abc
TRFK 6/8 1.315ab 3.830ab 0.6050
c 1.900ab
TRFK 31/8 1.485a 4.755efg 0.3400cdefg 0.8250efghi 1.450abc
TRFK 31/11 1.170abcde 2.375cde
0.8750defgh 1.255abc
TRFK 100/5 0.8150efgh 1.730efgh 0.4700
0.6900fghij 0.8900abc
TRFK 306/1
Means within a column followed by the same letter are not significantly different at p0.05 according to DMRT.
3.1.4 Gallic acid
The results on gallic acid (3, 4, 5-trihydroxybenzoic acid) are
presented in Table 4. The results show significant (p0.05)
differences in gallic acid content between the black (aerated),
green (non-aerated) and white (non-aerated) teas. This can be
attributed to the fermentation reaction where a considerable
quantity of EGC, EC, EGCG and ECG are oxidized to form
theaflavins and their gallates and gallic acid is an important
molecule in this reaction. Despite the formation of free gallic
acid during fermentation through the process of degallation
of EGCG, the enhanced utilization of gallic acid in the
formation of TFs and TRs contributes to the decline of gallic
acid though Muthumani and Kumar [30] in their study argued
that the decline is not significant. The formation pathways of
gallic acid have been shown to include the hydration of
epigallocatechin gallate and degradation from the dimer of
epigallocatechin gallate [31]. However, since these catechins
are vital in black tea formation, the hydration that leads to the
formation of gallic acid is therefore suppressed at the
expense of theaflavins and thearubigin formation. The levels
of gallic acid and individual catechins in the black tea have
been shown to decrease with an increase in fermentation
temperature and time for different clones [19, 20, 27].
Journal of Pharmacognosy and Phytochemistry
Table 4: Gallic acid (GA) (%) levels of different tea products
Clone Green Tea Black Tea
TRFK 301/4 1.075a 0.1350n
TRFK 301/5 0.4500jkl 0.1550mn
TRFK 303/216 0.5300ghi 0.2350jlk
TRFK 303/231 0.5250ghi 0.2900hij
TRFK 303/577 0.5900efg 0.3450fgh
TRFK 303/745 0.4650ijkl 0.3050ghi
TRFK 337/138 0.6250def 0.4250de
TRFK 371/3 0.4850hijk 0.2750ijk
TRFK 430/3 0.4900hij 0.2750ijk
TRFK 430/4 0.4100l 0.1700lmn
TRFK 430/12 0.4500jkl 0.1850lmn
TRFK 430/63 0.8650b 0.5350bc
TRFK 430/90 0.4800hijk 0.4600d
TRFK 524/170 0.7600c 0.4800cd
TRFK 524/48 0.7750c 0.3600efg
BBK35 0.8000bc 0.6050a
EPKC12 0.6650d 0.6250a
EPK D 99/10 0.4250jkl 0.1650mn
EJULU-L 0.8200bc 0.4750cd
AHPS 15/10 0.6750d 0.4600d
EPK TN 14/3 0.5650fg 0.210klm
TRFK 6/8 0.7800c 0.5950ab
TRFK 31/8 0.5450gh 0.3800ef
TRFK 31/11 0.5700fg 0.2800hijk
TRFK 100/5 0.6400de 0.3150fghi
TRFK 306/1
Means within a column followed by the same letter are not
significantly different at p0.05 according to DMRT.
3.1.5 Total theaflavins and total thearubigens levels of
black, green and white tea products
There was a significant difference in the total TFs and TRs
levels for Kenyan teas. Black tea had the highest levels of
total TFs and total TRs which ranged from 2.072% to
17.12%, respectively (Table 5). However, results from the
present study clearly showed that TRs were present in green
tea products and black and white (unaerated) tea products
from the purple coloured leaf tea clones. Further observations
revealed that in green and white teas, TRs were formed in the
presence of low levels of TFs unlike in black tea where the
TFs levels were slightly higher. Black tea therefore has high
levels of TFs and TRs that are the main fermentation
products as evident in table 5 below.
The variation in the polyphenolic composition of the
different tea products resulted from the leaf maceration
during manufacturing. The rolling and cutting of the tea
shoots in non-orthodox manufacture causes a release of
polyphenol oxidase which interacts with phenolic
compounds, one simple catechin and one gallocatechin, to
produce, theaflavins and thearubigins that possess a
benzotropolone skeleton [32, 33]. Owuor and Obanda [34]
investigated the use of green tea flavan-3-ols in predicting
black tea quality potential and revealed that a correct balance
of the trihydroxylated flavan-3-ols and dihydroxylated
flavan-3-ols was necessary to ensure maximum formation of
the theaflavins. The trihydroxylflavan-3-ols are oxidized
faster during the fermentation phase of black tea processing
explaining the high levels of EGCG and EGC in green tea
and the subsequent reduction in black tea. Theaflavins are
further oxidized to form thearubigins that are heterogeneous
in nature and contribute significantly towards taste, color and
body of tea [13, 19 & 35].
Results from the present study however clearly showed that
TRs were present in green tea. Further observation revealed
that in green tea, TRs were formed in the presence of low
levels of TFs unlike in black tea where the levels were almost
similar. This may suggest that theaflavins are not the only
source of thearubigins. Wilson and Clifford [36] explained the
factors affecting the formation and degradation of theaflavins
and thearubigins in black tea and observed that maximum
synthesis of theaflavins occurs when oxygen is in excess to
support benzotropolone ring formation. However, under
limiting oxygen concentration, polyphenol oxidase, which
has a high affinity for the substrate, has a preferential
demand for oxygen and theaflavins formation is suppressed
at the expense of catechin quinone formation. This
competition for oxygen is particularly noticeable during the
early stages of fermentation when the concentration of the
catechins is at its highest and enzyme turnover is unimpeded
by substrate availability. This occurs during green tea
manufacture since the enzyme is active before deactivation
through steaming. For this reason, high enzyme activity in an
already low oxygen concentration creates almost total
anaerobiosis, which suppresses benzotropolone ring
formation. Consequently as a result of this, thearubigens are
formed, mainly from gallocatechins since the simple
catechins are unable to react in benzotropolone ring
formation. Moreover, it might be possible to minimize
thearubigins formation by deactivating the enzyme
immediately after plucking through a steaming procedure
although this is hardly achievable during commercial tea
processing. Further research is desirable to explain in details
the existence of these thearubigins in green tea and the
importance of steaming during tea processing [14, 19, 26, 37, 39, 40].
Journal of Pharmacognosy and Phytochemistry
Table 5: Total theaflavins and total thearubigins (%) levels of black, green, purple and white tea products processed from different cultivars
Clone Green Tea Black Tea
TRFK 301/4 0.8500cde 9.705de
TRFK 301/5 0.6750defghij 10.02cde 1.555jkl
TRFK 303/216 0.5950efghij 9.775cde
TRFK 303/231 0.6500efghij 8.700hi 1.665defg 13.680hijkl
TRFK 303/577 1.025
c 11.48
1.525klmn 15.16
TRFK 303/745 0.8000cdefgh 9.835cde
1.655efgh 13.41fghij
TRFK 337/138 0.6800defghij 8.880hi 1.800
TRFK 371/3 0.5750fghij 12.15a 1.685cde
TRFK 430/3 0.6550defghij 6.935
TRFK 430/4 1.685a 9.74de
c 16.02ab
TRFK 430/12 0.8450cde 8.865ij 1.395
TRFK 430/63 0.5500ghij 8.68hi 1.475no 12.74ij
TRFK 430/90 0.6850defghij 9.280fgh 1.605ghij 13.22ghij
TRFK 524/170 0.4800ij 9.800cde
TRFK 524/48 0.5350hij 10.14cde 1.415op 14.00efghi
BBK35 0.6200efghij 8.980ghi 1.590hij
EPKC12 0.8100cdefg 10.18c
EPK D 99/10 0.9200
12.00ab 1.695cde 17.12a
EJULU-L 1.135
10.37c 1.735
c 16.03ab
AHPS 15/10 0.7200defghi 10.17c
1.505mn 13.40fghij
EPK TN 14/3 0.8250cde
1.515lmn 12.72ij
TRFK 6/8 1.150
12.28a 2.072a 16.13ab
TRFK 31/8 0.6750defghij 9.600de
1.470no 14.28defgh
TRFK 31/11 0.7350defghi 9.535efg 1.625fghi 13.90efghi
TRFK 100/5 0.4450j 8.05j 1.580ijkl 13.86efghi
TRFK 306/1
Means within a column followed by the same letter are not significantly different at p0.05 according to DMRT.
3.1.6 Theaflavin fractions
Theaflavins present in the assayed samples were fractionated
and found to contain the following fractions; theaflavin-3-
monogallate, theaflavin-3’-monogallate b, theaflavin-3, 3’-
digallate and theaflavin-3, 3- digallate. These fractions were
significantly different (p< 0.0001) in all tea cultivars. The
fractions differ in structure and previous studies have
elucidated them as shown in Figure 3.
Fig 3: Molecular structure of theaflavins. theaflavin (TF): R’= R’’=
OH, theaflavin-3-gallate (TF-3-G): R’=H, R’’= galloyl; theaflavin-
3’-gallate (TF-3’-G): R’ galloyl, R’’ H; theaflavin-3, 3’-digallate
(TF-dG): R’=R’’=galloyl.
Green tea catechins are oxidized and dimerized during the
manufacture of black tea to form orange red pigments
namely theaflavins (TF), which is a mixture of theaflavins
(TF1), theaflavin-3-gallate (TF2A), theaflavin-3’-gallate
(TF2B) and theaflavin-3,3’-digallate (TF3). These molecules
have recently aroused a lot of interest since they are thought
to have stronger biological properties than free theaflavins
due to the presence of gallic acid residues. In this study, the
theaflavin fractions correlated significantly well with each
other (Table 6) and this might explain why the activity of
black teas observed could not be entirely attributed to the
presence of theaflavins alone. The strong correlation of the
digillate fractions is a clear indication that they synergize
with other polyphenols to enhance the bioactivity. Obanda et
al., [20] observed that theaflavin gallate is the most astringent
and has been estimated to be 6.4 times more astringent than
simple theaflavin, and 2.88 times more astringent than either
theaflavin-3-monogallate or theaflavin-3’-monogallate. The
formation of a single theaflavin molecule requires a
dihydroxy and a trihydroxy flavan-3-ol, as follows:-
Epicatechin (EC) + Epigallocatechin (EGC) = simple
theaflavin (TF); EC + Epigallocatechin gallate (EGCg) =
Theaflavin-3-gallate (TF-3-g); Epicatechin gallate (ECG) +
EGC = Theaflavin-3’-gallate (TF-3’- g);ECG + EGCg =
Theaflavin-3, 3’-digallate (TF dg) [14]. Since theaflavin
fractions are proving to be essential biologically active
molecules, it is important to understand how the correct ratio
Journal of Pharmacognosy and Phytochemistry
of dihydroxyl flavan-3-ols to trihydroxyl flavan-3-ols can be
utilized to produce value added teas with enhanced quality
and biological use.
Table 6: Correlation coefficient matrix analyses between various
theaflavin fractions
1.000 0.962*** 0.873*** 0.961*** TF3MG
1.000 0.823** 0.908*** TF3’MG
1.000 0.937*** TF33DG
1.000 TF33’DG
** - Correlation significant at the p< 0.01 level
*** - Correlation significant at the p< 0.001 level
3.1.7 Antioxidant activity of green, black, purple and
white tea products
The polyphenolic composition of tea and especially its
catechins has aroused interest in their potential as radical
scavenging compounds. Data on antioxidant capacity is
presented in Table 7. Overall, green and white teas’ had
significantly (p< 0.05) higher antioxidant activity compared
to black tea. However, some black teas from cultivars such as
Ejulu-L, TRFK 6/8 and TRFK 306/1 had a higher antioxidant
activity compared to some unaerated green teas.
Table 7: Percent antioxidant capacity (AA) of green, black and
white tea products analyzed
Clone Green Tea Black Tea
TRFK 301/4 89.80defgh 87.55de
TRFK 301/5 88.30h 84.10
TRFK 303/216 90.55cdefgh 88.90c
TRFK 303/231 90.75cdefgh 89.15
TRFK 303/577 90.85cdefg 89.65abc
TRFK 303/745 90.55cdefgh 88.75c
TRFK 337/138 91.25cde
TRFK 371/3 91.35cde
TRFK 430/3 91.70
cde 88.45c
TRFK 430/4 89.10fgh 88.70c
TRFK 430/12 92.05abcde 88.80c
TRFK 430/63 88.65gh 86.00e
TRFK 430/90 89.60efgh 89.10
TRFK 524/170 91.90abcde 89.55abc
TRFK 524/48 91.15cdefg 89.60abc
BBK35 91.70
cde 89.15
EPKC12 91.90abcde 88.90c
EPK D 99/10 91.90abcde 88.55c
EJULU-L 94.05ab 91.10a
AHPS 15/10 92.50abc 89.80abc
EPK TN 14/3 89.00fgh 89.60abc
TRFK 6/8 94.30a 91.05a
TRFK 31/8 92.20abc
TRFK 31/11 92.45abc 89.30
TRFK 100/5 91.40cde
TRFK 306/1
+Data has been arcsine transformed.
Means within a column followed by the same letter are not
significantly different at p0.05 according to DMRT.
Table 8 presents data on the correlation between tea
polyphenols contents and the antioxidant activity of different
types of tea products. Total catechins significantly (p<0.001)
correlated with antioxidant activity (r=0.909***). EGCG was
identified as the most potent antioxidant (r=0.968***,
p<0.001). EGC (r=0.659***), EC (r=0.780***), ECG
(r=0.454*) and GA (r=0.530*) contents also showed
significant influence on the antioxidant activity. Therefore,
the antioxidant activity was higher in tea extracts containing
high levels of EGCG, EGC, and ECG. These results are
similar to those of Gramza et al., [41] and Karori et al., [5].
This antioxidative effect of polyphenols has been attributed
to the phenolic hydroxyl groups in their structures that make
them potent free radical scavengers [42]. The antioxidative
properties of catechins are marked particularly by their
ability to inhibit free radical generation, scavenging free
radicals and chelate transition metal ions mainly, iron and
copper [43]. Nakagawa and Yokozawa [44] showed in their
study that green tea extracts significantly impaired nitrogen
oxide production in a concentration dependent manner and
showed a direct scavenging activity against super oxide
anion. On the basis of these results, it appears that the most
effective radical scavengers are catechins with a 3’, 4’ and
5’-trihydroxylated substitution pattern on the B ring and/or
hydroxyl group at C-3 position of the catechin structure
(Figure 3). This hydroxylation confers a higher degree of
stability on the catechin phenoxyl radical by participating in
electron delocalisation that is an important feature of the
antiradical potential. A study using electron spin resonance
showed that the presence of 3’, 4’, and 5’-trihydroxyl groups
attached to the B-ring of the flavan skeleton enhanced the
radical scavenging efficiency displayed by catechins,
compared to those with 3’, 4’-dihydroxyl groups. At the
same time the insertion of a galloyl moiety into three
positions of the C-ring exerted a synergistic impact on
superoxide anion scavenging activity [43, 45]. This explains
why radical scavenging is high in the gallocatechins namely
EGCG and EGC that are potent antioxidants [42, 46, 47].
To support this observation that EGCG and ECG were potent
antioxidants, the findings correlated well with a study
conducted in the Republic of Korea that showed that EGC
has the highest specific total oxy-radical scavenging capacity
against peroxyl radicals, hydroxyl radicals and peroxynitrite,
while ECG was the least effective among other catechins [48].
Raza and John [49] reported that tea catechins prevent
molecular degradation in oxidative stress conditions by
directly altering the subcellular ROS production, glutathione
metabolism and cytochrome P450 2E1 activity. These results
are promising for the chemotherapeutic use of tea catechins
in oxidative stress-related diseases.
Black teas analyzed in this study exhibited some antioxidant
activity with a high DPPH radical scavenging activity though
less than that of green, white and purple teas. During black
tea manufacture, the gallocatechins are first to be oxidized
and dimerised to TFs and TRs because of their high
oxidation potential and high concentration in the leaves.
These major phenolic compounds in black tea also
contributed significantly to the radical scavenging activity
namely TFs (r=0.803***, p<0.001), TRs (0.859***,
p<0.001) and GA (r=0.530*, p<0.05). Interestingly, TFs,
which are the major phenolic products in black tea, had a
higher radical scavenging activity compared to some of its
precursors ECG, EGC and EC (Table 8). This confirms that
Journal of Pharmacognosy and Phytochemistry
conversion of catechins to TFs during black tea process did
not affect the radical scavenging potency. These observations
are consistent with those of Leung et al. [21]; Karori et al., [5]
and Wachira et al., [12] who showed that black tea posses
more or less the same antioxidant potency as catechins
present in green tea. EGCG and EGC contribute significantly
to the formation of TFs. These are B ring trihydroxylated
catechins, which are oxidized at a much faster rate than the B
ring dihydroxylated catechins including EC, ECG and +C
due to their lower oxidation potential [24]. TFs formed from
this reaction have hydroxyl groups (OH) considered
necessary for free radical scavenging activity. These
additional groups increase the total number of phenyl
hydroxyl groups and make the gallate containing catechins
and TFs more able to donate protons due to resonance
delocalization thereby expressing the observed antioxidant
activity of black tea. Similarly, gallic acid contributed
significantly to the radical scavenging activity in black tea
because it is a potent hydrogen donator to DPPH.
Table 8: Correlation coefficient matrix analyses between various tea chemical parameters
1.00 0.818*** 0.663*** 0.718*** 0.803*** 0.715** 0.520** 0.626** 0.895*** 0.830*** 0.681*** TP
1.00 0.791*** 0.732*** 0.852*** 0.632* 0.452* 0.584* 0.803*** 0.808*** 0.689*** TFs
1.00 0.686** 0.843*** 0.619** 0.393* 0.378* 0.859*** 0.826*** 0.733*** TRs
1.00 0.847*** 0.505** 0.291* 0.552** 0.659*** 0.856*** 0.688*** EGC
1.00 0.719** 0.444*** 0.656** 0.968*** 0.956*** 0.777*** EGCG
1.00 0.744* 0.637*** 0.454* 0.8300*** 0.814*** ECG
1.00 0.472 0.232* 0.608*** 0.690*** +C
1.00 0.530* 0.685** 0.628** GA
1.00 0.909*** 0.780*** AA
1.00 0.895*** TC
1.00 EC
Additionally, the present study provided evidence of the
contribution of TRs towards the antioxidant activity of black
tea (r=0.803***, p<0.001). The antioxidant activity of TRs
can be explained by the presence of 3-OH groups, which are
more or less esterified by gallic acid in the TRs structure.
However, this is a highly speculative hypothesis since to date
there is no definite data on TRs structures [21, 50].
Thearubigins are assumed to be formed by the tea plant as a
defense mechanism [35]. Plants are thought to utilize the
strategy of plant browning as a defense tool. Therefore, the
action of a polyphenol oxidase enzyme on phenolic
secondary metabolites to produce a brownish coloration is
aimed at deterring pest organisms. The tea plant uses a
similar process to oxidize flavan-3-ols to thearubigins using
tea polyphenol oxidase to gain an evolutionary advantage by
deterring pest organisms [51]. Thearubigins account for
around 60% to70% of the dry weight of a typical black tea
infusion, and any attempt to understand the numerous
beneficial health effects of this beverage must take this class
of compounds into consideration. However, only a few
studies on the biological effects of thearubigins are available.
The reason for this lack of knowledge is obvious because
TRs have been mysterious for decades, with no clear
structural picture and only vague and sometimes
contradictory knowledge available. Therefore, there are no
defined compounds from the TR fractions that can be used
for biological testing, hence no standardized method for
obtaining extracts for biological testing. Despite these
limitations, TRs are useful molecules that need a detailed
study to establish their role in disease prevention. An attempt
so far made in this field has established that TRs extracts
lowered the expression of superoxide dismutase, a free
radical scavenger, in contrast to theaflavins, TR extracts were
able to inhibit DNA synthesis in U-937 leukemia cell lines,
giving a rationale for the anti-cancer activity of TRs [52]. Lin
et al., [3] showed that TR extracts were able to block nitric
oxide synthase in macrophages and therefore suppress the
anti-inflammatory response and multiple stages of
carcinogenesis. Data obtained from this study reveals that
different tea cultivars have different polyphenolic
composition which imparts on their unique biochemical
4. Acknowledgement
We acknowledge the Tea Research Institute (TRI) and the
Division Research and Extension-Egerton University.
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... Phytochemicals and functional components in tea are receiving a lot of attention due to their potential benefits in health when consumed as part of a varied diet on a regular basis and at effective levels. Many nutraceuticals, functional foods and naturally occurring polyphenols have physiological and pharmacological activities including their well characterized antioxidant properties [1][2][3]. The scientific and food industry communities share a common goal of extending the quality of human life by utilizing phytochemicals. ...
... Tea is rich in polyphenols associated with numerous pharmacological properties such as; antimicrobial [5], anti-inflammatory [6], anti-aging [7,8] antimalarial [9] and antioxidant activity [1,3,10]. Cancer, accelerated aging and other chronic disease associated with free radical induced oxidative damage [4] can be ameliorated by phytochemicals in tea since antioxidants in tea have been shown to protect DNA from damage induced by free radicals and slow or halt initiation and progression of cancerous tumor growth [12]. ...
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Background Tea (Camellia sinensis) infusions are widely consumed beverages with numerous health benefits. However, physiological and molecular responses mediating these activities are poorly understood. Method Three replicates of 4TI cancer cell suspension (2.0 × 105 cells/ml) were challenged in vitro with various concentrations of green, black and purple tea infusions to asseses their cytoxicity and associated differentially expressed genes in the cells. Inhibitory activity was tested by using serial dilutions of respective tea infusions in a 96 well ELISA plate. ResultsGreen tea had the highest inhibition on 4TI cells proliferation at a concentration of IC50 = 13.12 μg/ml. Further analysis of the 4TI cancer cell line treated with tea using 454 pyrosequencing generated 425,696 reads with an input mean length of 286.54. Trimmed sequences were imported on a CLC genomic workbench v7.03 and annotated on a reference mouse genome (Mus musculus strain C57BL/6 J). Results revealed a differential expression of apoptosis related genes in the transcriptome. Casp8, Casp9, Casp3, Casp6, Casp8AP2, Aifm1, Aifm2 and Apopt1 genes were significantly upregulated indicating the process of apoptosis was initiated and executed. Conclusion These findings on caspases offer valuable information on the mechanism of tea as an anticancer agent and will contribute to further research in future novel treatments.
... Green tea catechins, black tea theaflavins, and caffeine are important biochemical traits that determine tea quality given that tea price at the auction is based on tea quality. The two principal gallated catechins present were EGCG and EGC which corroborate with results reported by (Karori et al. 2014). The two gallated catechins (EGCG and EGC) have been shown to contribute significantly to the formation of theaflavins. ...
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Tea (Camellia sinensis) contains polyphenols and caffeine which have been found to be of popular interest in tea quality. Tea production relies on well-distributed rainfall which influence tea quality. Phenotypic data for two segregating tea populations TRFK St 504 and TRFK St 524 were collected and used to identify the quantitative trait loci (QTL) influencing tea biochemical and drought stress traits based on a consensus genetic map constructed using the DArTseq platform. The populations comprised 261 F1 clonal progeny. The map consisted of 15 linkage groups which corresponds to chromosome haploid number of tea plant (2n = 2× = 30) and spanned 1260.1 cM with a mean interval of 1.1 cM between markers. A total of 16 phenotypic traits were assessed in the two populations. Both interval and multiple QTL mapping revealed a total of 47 putative QTL in the 15 LGs associated with tea quality and percent relative water content at a significant genome-wide threshold of 5%. In total, six caffeine QTL, 25 catechins QTL, three theaflavins QTL, nine QTL for tea taster score, and three QTL for percent relative water contents were detected. Out of these 47 QTL, 19 QTL were identified for ten traits in three main regions on LG01, LG02, LG04, LG12, LG13, and LG14. The QTL associated with caffeine, individual catechins, and theaflavins were clustered mostly in LG02 and LG04 but in different regions on the map. The explained variance by each QTL in the population ranged from 5.5 to 56.6%, with an average of 9.9%. Identification of QTL that are tightly linked to markers associated with black tea quality coupled with UPLC assay may greatly accelerate development of novel tea cultivars owing to its amenability at seedling stage. In addition, validated molecular markers will contribute greatly to adoption of marker-assisted selection (MAS) for drought tolerance and tea quality improvement.
... Thearubigins, the orange brown compounds, are responsible for the colour and taste of tea. Thearubigins contribute approximately 35% of total colour, and also play a significant role in the brown colour of tea, as well as in the strength and mouth feel of the tea liquor (Kumar et al. 2011, Karori et al. 2014. ...
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Tea (Camellia sinensis) is an important beverage crop cultivated in the tropics and subtropics under acid soil conditions. Increased awareness of the health-promoting properties of the tea beverage has led to an increase in its level of consumption over the last decades. Tea production contributes significantly to the economy of several tea-cultivating countries in Asia and Africa. Environmental constrains, particularly water deficiency due to inadequate and/or poorly distributed rainfall, seriously limit tea production in the majority of tea-producing countries. It is also predicted that global climate change will have a considerable adverse impact on tea production in the near future. Application of fertilizers for higher production and increased quality and quantity of tea is a common agricultural practice, but due to its environmental consequences, such as groundwater pollution, the rate of fertilizer application needs to be reconsidered. Cultivation of tea under humid conditions renders it highly susceptible to pathogens and pest attacks. Application of pesticides and fungicides adversely affects the quality of tea and increases health risks of the tea beverage. Organic cultivation as an agricultural practice without using synthetic fertilizers and other chemical additives such as pesticides and fungicides is a sustainable and eco-friendly approach to producing healthy tea. A growing number of tea-producing countries are joining organic tea cultivation programmes in order to improve the quality and to maintain the health benefits of the tea produced.
... In this study, the sampled cultivars contained EGCG fraction in largest amounts followed by EGC and ECG, while +C and EC were present in significantly lower amounts. Similar results were obtained in studies done by Karori et al 42,43 and Ender et al 44 . Correlation analysis showed a significant ( contribution of the individual catechin fractions to the TC content with EGC contributing the most (r=0.600***) ...
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Tea industry in Kenya contributes to the economy by being the largest agribusiness and a major foreign exchange earner. However, significant revenue is lost when tea is sold in undiversified form. This has created a need to characterize the available clones for suitability in high value tea product development. Samples were obtained from 204 tea accessions conserved in Kericho and Kangaita Centres of the Tea Research Institute (TRI) and assayed for total polyphenols, catechins, caffeine, anthocyanins, chlorogenic acid and theanine. There were significant cultivar differences (P≤0.05) in total polyphenols (16.4%-30.9%) and total catechins (11.03%-25.42%). Sixteen (16) new clones recorded significantly (P≤0.05) higher polyphenol contents (mean value of 28.11%) than the standard reference clone, TRFK 6/8 (27.4%) indicating their suitability in the development of high quality black teas. Fifteen clones were suitable for the manufacture of theaflavin-3, 3'-digallate rich black tea based on their high ECG and EGCG levels. Clones TRFK 301/5 and TRFK 301/4 had a high EGC/EC and low EGCG/ECG ratios and were found suitable for manufacture of less astringent green orthodox teas, while clones TRFK 687/1 and 73/7 had the least caffeine contents at 1.96% and 2.04%, respectively, implying their amenability for manufacture of low-caffeine tea beverages. Clones assayed for chlorogenic acid and theanine showed that AHP SC 31/37 and TRFK 6/8 had the highest contents at 0.13% and 1.7%, respectively, and are suitable for chlorogenic and theanine rich teas. The observed chemical and therefore quality differences based on clones and regions show that these Kenyan tea cultivars have high diversity in biochemical attributes and maybe suitable for development of diversified tea products with geographical indications.
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Tea (Camellia sinensis L.) is the most widely-consumed beverage in the world. The biochemical components of tea leave include polyphenols (catechins and flavonoides), alkaloids (caffeine, theobromine, theophylline, etc.), volatile compounds, polysaccharides, amino acids, lipids and vitamins show a variety of bioactivities. Prolong cross-pollination nature of tea plants have produced considerable heritable variation, resulting in a high level of genetic diversity. The collection and conservation of the cultivars, landraces and wild relatives of the tea plant provides breeders with fundamental materials from which new cultivars are to be developed. The major role of tea breeding is to improve productivity, enhance tolerant to biotic and abiotic stress, and increase tea flavor and quality. Dissection of the genetic basis of these traits provides the potential for accelerating the breeding process by developing new tools such as marker-assisted selection. Therefore present review provides an overview of the biochemical and metabolite diversity of the global tea germplasm and its characterization and utilization.
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In the present study, twelve different types of commercial tea samples were assayed to determine their phenolic composition and antioxidant activity. Reverse phase high performance liquid chromatography using a binary gradient system was used for the identification and quantification of individual catechins. Subsequently, total phenolic content was determined spectrophotometrically according to the Folin-ciocalteus method. Total theaflavins and thearubigins were also determined. The radical scavenging behavior of the polyphenols on 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) was also studied spectrophotometrically. The results showed that total polyphenols, total catechins and antioxidant activity were significantly (P<0.05) different in the commercial tea samples. Green tea had the highest levels of catechins, total polyphenols and total antioxidant activity. White tea (silvery tip) a rare specialty type of tea was not significantly different from green tea. Statistical analysis showed an essential catechin content influence of the tea extracts on antioxidant activity. Epigallocatechin gallate (EGCG) was the most potent catechin and the most potent in antioxidant activity (r = 0.989***). Epigallocatechin (EGC) (r = 0.787, P<0.001), epicatechin (EC) + catechin (+C) and epicatechigallate (ECG) also showed significant (P<0.05) antioxidant activity. Black tea contained high levels of theaflavins and thearubigins, which accounted for most of the antioxidant potential in this type of tea product (r = 0.930*** and r = 0.930*** respectively). These results suggest that conversion of catechins during black tea processing did not affect the free-radical potency of black tea. Gallic acid (GA) also showed significant(r = 0.530*) contribution to the antioxidant activity in black tea. Green, black and white tea products processed from Kenyan tea cultivars originally selected for black tea had significantly (P<0.05) higher antioxidant activity than green tea processed from tea cultivars from Japan and China. These results seem to suggest that the cultivar type is critical in determining the antioxidant potency of tea product and that black teas processed from suitable cultivars could be potent in antioxidant activity when compared to green teas.
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The relationship between the structural characteristics of 29 flavonoids and their antiradical activity was studied. The obtained results suggest that the free radical scavenger potential of these polyphenolic compounds closely depends on the particular substitution pattern of free hydroxyl groups on the flavonoid skeleton. The possible mechanism of action of flavonoids lacking B ring OHs as free radical scavengers has been proposed.
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Green tea is sourced from Camellia sinensis (L.) O. Kuntze (Theaceae), the same species from which white, oolong, black and pu-erh teas are derived. The various tea types are classified on the basis of their processing, and the associated oxidation and fermentation levels which influence taste and aroma profiles. Green tea is a minimally oxidized and non-fermented tea. Catechin polyphenols are the primary compounds responsible for the claimed health benefits of green tea, including its antioxidant and anti-inflammatory properties. Caffeine contributes to green tea's stimulant properties, while the amino acid theanine contributes to its relaxing properties. Significant variation of catechin and caffeine content occurs between green tea types depending on environmental growth conditions and processing. Most often, green teas are sourced from the sinensis variety of the tea plant while black teas are sourced from the assamica variety. China is the center of origin of green tea production. Today, green tea is produced in over 20 countries in tropical, sub-tropical and temperate regions. It is the most widely consumed beverage after water, due to its health, sensory, stimulant, relaxing and cultural properties. The quality of green tea is impacted by numerous factors involved in cultivation, harvest, processing, storage and preparation that influence chemistry, taste, aroma, morphology and bioactivity of tea leaves. s0015 THE PLANTS: BOTANICAL CLASSIFICATION AND DISTRIBUTION s0020 Botanical Identification p0015 The tea plant was first described taxonomically in 1753 by Carl Linnaeus in Species Plantarum. He referred to the tea plant as Thea and later refined the species into black tea (Thea bohea) and green tea (Thea viridis). By the early 1900s, taxonomists recognized that green and black tea were both from the same species, Camellia sinensis (L.) O. Kuntze. p0020 Camellia sinensis is an evergreen tree or shrub. It has yellow-white flowers and long, serrated leaves. Flowers are axillary, solitary, or up to three in a cluster. They are 2.5e3.5 cm in diameter and have six to eight petals. The outer petals are sepaloid and the inner petals are obovate to broadly obovate. There are numerous stamens 0.8e1.3 cm in length. Young leaves have short 19
The black tea manufacturing process, as described in Chapter 14, is instrumental in disrupting the cellular integrity of green tea shoots and, in so doing, facilitates the mixing of polyphenols held in the cell vacuole with cytoplasmic polyphenol oxidase. The biochemical and chemical reactions which then follow, result in the uptake of atmospheric oxygen and the formation of the pigmented, hot water-soluble, polyphenolic compounds, characteristic of black teas.
White tea is a rare tea that is derived from the same plant ( Camellia sinensis ) as green, oolong and black tea. Originating from and predominantly produced in southern China, it was virtually unknown to the western world until the late 1800s. Primarily consisting of the silvery buds and young leaves of the plant, it is hand-picked, steamed and dried without further, elaborate processing, which accounts for the whitish appearance. It is the least processed of all tea. The minimal processing is believed to result in the preservation of high amounts of phytochemicals that confer health benefits and its advantage over other types of tea. This chapter provides a general overview of the origin, the major varieties, processing, manufacturing and production of white tea. A summary of published studies that support the potential health promoting benefits of white tea, including its anticancer, antiobesity, antioxidant and antiaging properties is also provided.
Clonal differences and fermentation conditions have an impact on black tea in terms of unoxidized catechins and antioxidant activity. During processing, black teas from different clones differ in their levels of gallic acid, individual catechins and antioxidant activity. Dependant on the processing conditions of the black tea clones, the levels of gallic acid, epigallocatechin gallate (EGCG), total polyphenols and consequently antioxidant activity vary significantly. Gallic acid, (+)-catechin, EGCG and total polyphenols are the major contributors to black tea antioxidant activity. The black teas processed show more than 70% inhibition of α,α-DPPH radical solution. Optimal clone-specific processing conditions can therefore be utilized to add value to black tea with regard to antioxidant activity.
Tea has been considered a palatable beverage for over 4,000 years before the scientific community advocated its therapeutic potential. It is a complex mixture containing a range of compounds from simple phenolics to complex tannins, many of which have well-recognized biological properties. Epidemiological and experimental investigations have provided evidence that tea, in all its three major forms (green, black and oolong), may exert a retarding effect on the etiology and development of diabetes mellitus, cancer, cardiovascular diseases and a number of degenerative diseases. The prophylactic effects of tea have been mainly attributed to their polyphenolic compounds, which have been comprehensively characterized. In addition, several schools of thought have linked tea health claims to its antioxidant effectiveness, as assessed using a wide range of assays. However, tea variety, weight of tea or teabag, brewing techniques, and solvent systems have all been shown to affect greatly extraction efficiencies of tea polyphenols. It remains unclear which system is more effective for optimum extraction and evaluation of antioxidant activity. This chapter reviews some of the literature data on the phytophenolic contents and antioxidant functions of aqueous and organic tea extracts.