E. T. Contis et al.

(Editors)
Food Flavors: Formation, Analysis and Packaging Influences
© 1998 Elsevier Science B.V. All rights reserved 423

Characterization of f l a v o r of t e a p r o d u c e d d i f f e r e n t tea area

Miyuki K A T O ' a n d M a s a s h i OMORl'

^Faculty of Education KAGAWA University. T a k a m a t s u KAGAWA 760 J a p a n .

^Department of Food Science Otsuma Wumen's University. 12 Sanban-cho

Chiyoda-ku Tokyo 102 J a p a n

Ab 8 t r a c t
India, Sri Lanka and China are major, well known tea producing a r e a s . In
recent years, the production of Kenian and I n d o n e s i a n tea also have been
increased. In this study, a u t h o r s were comparatively found out about flavor
components of these t e a s produced different tea a r e a . (l)It was showed high
contents of polyphenol in Kenian tea, high contents of catechins in A u s t r i a n and
Kenian t e a . (2)The correlations among the scores (axes 1 and axes 2) obtained
by the quantification system III and p e a k s in the gaschromatograms obtained
by analyzing the flavor components were determined. The correlation coefficients
of the Axes 1 and 2 showed t h a t possible to do group with the flavor components
in each black tea s a m p l e .

1.INTRODUCTION

Well-known black tea-producing a r e a s include Darjeeling and Assam in

I n d i a , K e e m u n in China, Uva,Dimbula and Nuwara Eliya in Sri L a n k a . A
number of studies have been performed on the aroma components and coloring
components of these black tea products. Black tea products grown in Indonesia
and Kenya have been frequently brought to the m a r k e t in recent years. There
h a s been an increase in the production and exportation of black tea in these
a r e a s and Southeast Asia. Gianturco et al. and Y a m a n i s h i et al. identified a
number of aroma components [1-3]. On the black tea while Takino et al.[4]
determined the structure of theaflavin (TF) as a coloring component. Many
workers have studied the structure of t h e a r u b i g i n (TR), however no r e m a r k a b l e
finding has been e s t a b l i s h e d since Roberts et al. used the name "TR" to refer to
a mixture of compounds[5,6].
Since information of black tea aroma compounds formation is lacking, we
studied the formation mechanism of black tea aroma components by using tea
species grown in J a p a n . We have reported t h a t /3 -carotene plays an i m p o r t a n t
role in the formation of the a r o m a [ 7 , 8 ] . In this study, we have p r e s e n t clarify
information to the flavor composition of black tea products other t h a n those
obtained in the m a i n producing a r e a s . We have performed m u l t i v a r i a t e
analysis(MVA) on aroma components, the MVA was able to d e m o n s t r a t e d
differences in aroma components t h a t were dependent the producing a r e a s .
424

2.MATERIAL AND M E T H O D S

2.1.Samples
The s a m p l e s utilized were black t e a s produced in Kenya, Turkey,
Bangladesh, Myanmar, Viet Nam and A u s t r a l i a .

2.2. Sensory test

Sample were t e s t e d for quality using the tea evaluation method[9]. Two
hundred mL of boiling w a t e r was added to 3g of each sample in a cup, and
allowed to s t a n d for 3 min. The extract was filtered and then e v a l u a t e d . The
sensory t e s t s were carried out and discussed by skilled p a n e l i s t s .

2 . 3 . D e t e r m i n a t i o n a n d a n a l y s i s of b l a c k t e a e x t r a c t color
Black tea extracts were prepared in the same m a n n e r as follows. One
hundred mL of each extract was re-extracted with the same amount of methyl
isobutyl ketone (MIBK). The MIBK layer was s e p a r a t e d from the aqueous layer.
The aqueous layer was further extracted with the same amount of butanol
(BuOH), and the BuOH layer was s e p a r a t e d . Absorption of each extract at
380nm was recorded.
The polyphenol content was determined by the method of I w a s a et al [10].
The content of theaflavin was determined by the method of Anan et al [11]. For
theaflavin, 1 g of black tea leaves was extracted with 50 mL of 20% acetone
containing 0.6 mg of puropurogallin for 1 h; the volume of the acetone layer was
brought to 100 mL, High performance liquid chromatography (HPLC) was
performed with Innersil ODS-2 column (10 /xm, Gasukuro kogyo,4.6X250mm) for
a n a l y s i s of theaflavin as described below.

2 . 4 . A n a l y s i s of a m i n o a c i d s
Amino acids were analyzed according to the method of T s u s h i d a et al [12].,
in which 1 g of sample was extracted with 50 mL of boiling w a t e r for 60 min.
The resulting m a t e r i a l was made up to 100 mL with distilled water. The
extract was p a s s e d through a 0.45 /x m millipore m e m b r a n e filter, and
analyzed with a Shimadzu HPLC model LC-6A amino acid analysis system,
using the o-phthalaldehyde method.

2 . 5 . A n a l y s i s of c a t e c h i n s
Cathechins were analyzed according to the method of Ikegaya et al [13]. In
this method,100 mg of sample was extracted with about 30 mL of 20% acetone
for 60 min and filtered. The filtrate was extracted with about 100 mL of ethyl
a c e t a t e before condensing. The remaing m a t e r i a l was made up to 10 mL with
25% acetonitrile. The extract was p a s s e d through a 0.45 M m millipore
m e m b r a n e filter and t h e n injected into the Hitachi L-6200 i n s t r u m e n t .
 425

2 . 6 . A n a l y s i s of a r o m a c o m p o n e n t s
Black tea s a m p l e s were analyzed in accordance with the method of Horita
et ai[14]. Analysis was preformed by a s i m u l t a n e o u s distillation and
extraction (SDE) method, where distilled w a t e r was employed of a level 20
times t h a t of each s a m p l e . The d i s t i l l a t i o n was continued for 60 min.
Gas-chromatography (GC) was performed from 50-180°C a t a heating rate of 2V>
/min. A silica capillary column (0.35 mm i.d. X 50 m) coated with PEG-20M was
used. Flame ionization detection (FID) was used as a detector. The content of
components was expressed relative to the area of ethyl decanoate which was
used as an i n t e r n a l s t a n d a r d and identified using a Shimadzu GC-MS QP
lOOOA.

2.7. Multivariate a n a l y s i s method

Multivariate a n a l y s i s was performed by determining the area r a t i o of
the aroma components of each sample with the use of "Multi Tokei System"
m a r k e t e d by S h a k a i Joho Service. By using the area ratio of the aroma
components, the characteristic vector value of each factor was first calculated
and then the contribution ratio of the factor was calculated by using the vector
value. Thus, axes 1 and 2, were determined and served as the main
components for analyzing the vector p a t t e r n . For each tea sample, the factor
load was calculated and the vector p a t t e r n plotted by referring the ordinate
and the abscissa respectively as the first and second components.

3 . R E S U L T A AND D I S C U S S I O N

3.1. Morphological characteristic

The s a m p l e s of Kenya, Bangladesh, A u s t r a l i a , Turkey, Viet Nam and
Myanmar all had a round shape owing to the Crush Tear Curl(CTC) production
process.

3.2. C h a n g e s in c o l o r i n g c o m p o n e n t s of b l a c k t e a e x t r a c t s
Table 1 shows the composition of black tea coloring components, the
solvent fractionation method and the theaflavin contents. The black tea s a m p l e
of Kenya showed the g r e a t e s t content of the coloring components followed by
those of Bangladesh, A u s t r a l i a , Turkey, Viet Nam and Myanmar. The s a m p l e
of Sri L a n k a (Uva), showed the middle content of coloring compounds. It was
considered t h a t the coloring components should be easily eluted in the case of
the Kenya black tea which is t a k e n as milk tea in general. As the result of the
solvent fractionation method, the Kenya black tea showed the largest content of
the MIBK layer containing theaflavin (TF), while the samples of Turkey,
Myanmar and Viet Nam showed small contents. Compared with the Uva black
tea sample, the Kenya black tea sample showed a large content of the BuGH
426

layer containing thearubigin (TR) while the samples of other a r e a s showed

small contents thereof. All of the s a m p l e s showed small polyphenol contents,
though those of Kenya, Myanmar and Australia showed relatively large
polyphenol contents. When examined by HPLC, the Kenya black tea sample
showed a high TF content while those of Myanmar and Viet Nam showed s m a l l
ones, similar to the d a t a of the MIBK layer obtained by the solvent
fractionation method. The Kenya black tea sample showed a large TF content.

Table 1
Color comparison of black tea infusion (mg/lOOg dry matter)
Kenya Turkey Bangla- Myanmar Viet nam Aus- Sri
desh trallia Lanka
conponents
Infusion* 6.40 4.19 5.26 3.84 3.97 5.26 4.94
MIBK layer* 2.47 0.23 1.39 0.48 0.52 1.39 1.45
BuOH layer* 3.00 1.77 1.92 2.00 1.46 1.92 2.48
Polyphenol contents** 13.0 5.9 9.7 10.0 7.2 12.2 15.0
Contents of theaflavins***
Theaflavin 676.8 372.6 301.4 67.0 122.2 193.3 404.0
Theaflavin-3-monogallate 240.3 207.5 231.7 55.0 56.8 113.3 316.0
Theaflavin-3'-monogallate 221.0 213.5 182.8 56.2 69.4 127.1 237.0
Theaflavin-3-3'digallate 113.8 257.8 267.1 72.6 65.6 154.1 242.0
Black tea leaf was extracted with 200ml of boiling water for 3min.
*0D at 380nm, **%

3.3. C h a n g e s in a m i n o a c i d c o n t e n t s
Table 2 shows the amino acid contents of the black tea grown in various
region. The samples of each area were rich in t h e a n i n e , an amino acid
contained in a large amount in t e a . The samples of Bangladesh, Myanmar,
Australia and Kenya showed large total amino acid contents, while those of Viet
Nam and Turkey showed smaller total amino acids content. The s a m p l e s of
each area was rich in a s p a r t i c acid, serine and glutamic acid which are amino
acids contained in a large amount in tea Kato et al[15].

3.4. C h a n g e s in c a t e c h i n c o n t e n t s
Table 3 shows the results of catechin content determination. Compared
with the s a m p l e s of the major producing a r e a s such as Uva and Assam, the
black tea s a m p l e s employed in this study contained catechins in small a m o u n t s .
In particular, the samples of Turkey, Bangladesh and Viet Nam showed small
catechin contents. With respect individual catechin, these s a m p l e s were
characterized by being rich in (-)-epigarocatechin gallate [(-)-EGCg] and
(-)-epicatechin gallate [(-)-ECg].
 427

Table 2
Contents of amino acids in black tea (mg/lOOg dry matter)
Kenya Turkey Bangla- Myanmar Viet nam Aust Sri
desh trallia Lanka
Asp artic acid 284.8 147.7 170.5 146.5 101.9 160.5 24.8
Theronine 33.4 26.3 26,8 26.5 19.9 60.4 10.6
Theanine 824.0 622.1 1487.4 1043.2 960.4 1084.4 284.9
Glutamic acid 200.1 228.7 162.2 176.9 165.8 171.6 44.7
Isoleucine 10.5 17.2 16.6 24.9 16.9 17.1 3.2
Leucine 18.4 19.5 13.6 71.1 27.9 18.5 14.8
Tyrosine 54.4 40.5 11.5 43.9 25.7 31.9 20.8
Phenylalnine 26.4 176.1 50.0 66.2 54.4 26.6 34.4
Histidine trace 6.4 trace 6.7 5.1 trace 2.1
Lysine 77.5 14.1 0.2 16.8 4.9 trace 8.9
Arginine 61.0 49.5 17.2 40.9 30.0 41.9 2.6
HPLC instrument:Shimadzu LC-6A. 0 -phthalaldehyde method

Table 3
Contents of catechins in black tea (g/lOOg dry matter)
Kenya Turkey Bangra- Myanmar Vietnam Aust- Sri
desh rallia Lanka
(-)-EGC 0.722 trace 0.157 0.319 trace 0.282 0.193
(-)-EC 1.092 trace 0.020 trace 0.062 0.139 0.348
(-)-EGCg 1.900 0.076 0.331 1.118 0.451 0.852 5.123
(-)-ECg 1.994 0.075 0.312 1.199 0.444 1.135 3.342
Total 5.707 0.150 0.820 2.636 0.957 2.408 9.006
(-)-EGC:(-)-Epigallocatechin,(-)-EC:(-)-Epicatechin,(-)-EGCg:(-)-Epigallocatechingallate
(-)-ECg:(-)-Epicatechin gallate
HPLC instrument:Hitachi L-6200; coliumn:Hibar Lichrosorb RP-18(5Mm),
(^4.0mm X 250mm; column temp.:30°C;mobile phase;acetonitrile:acetic acid
:methanol:H2O(113:5:20:862);detector:280nm;flow rate:1.0ml

3.5. C h a n g e s in aroma components

T a b l e 4 shows the aroma volatile composition relative to ethyl d e c a n o a t e
as internal standard. Data indicated t h a t the black tea s a m p l e s in these
a r e a s contained less aroma components in general. Compared with t h e s a m p l e s
of the major producing a r e a s , t h e s e black tea s a m p l e s showed lower levels of
t h e t e r p e n e compounds c h a r a c t e r i s t i c to black t e a . Uva and Darjeeling black tea
s a m p l e s were rich in geraniol, benzyl alcohol and 2-phenyl e t h a n o l . However,
t h e black tea s a m p l e s employed in t h i s s t u d y contained t h e s e compounds only
in small amounts. These samples contained somewhat larger amounts of
components observed in green t e a .
428

Table 4
Composition of volatile flavor compound '
tR
(min) Compound Kenya Turkey Bangla- Viet n a m Aus- Sri
desh trallia Lanka
10?7 Hexanol L16 0.69 0.47 0^8 0.67 0.56
12.1 (Z)-3-Hexanol 1.25 0.61 0.27 0.66 0.45 2.51
15.9 Linalool oxide 0.24 0.46 0.62 0.35 0.39 0.39
(cis-furanoid)
18.4 Linalool oxide 0.50 0.59 0.90 0.15 0.39 1.89
(trance-furanoid)
21.0 Linalool 4.71 10.04 1.30 2.05 0.59 3.95
27.4 Eo-terpineol 0.60 A} trace A} 0.10 0.97
31.2 Linalool oxide pyranoid 0.60 0.78 0.28 0.57 0.17 0.26
32.1 M e t h y l salicylate 0.31 trace 0.52 A} 0.25 2.23
32.8 1-phenyl e t h a n o l 0.21 0.32 trace 0.14 0.03 0.45
37.1 Geraniol 1.26 1.26 0.70 0.36 0.53 2.36
38.4 Benzyl alcohol 0.13 1.77 0.87 0.41 0.16 3.34
42.0 2-phenyl e t h a n o l 18.63 0.06 5.60 1.73 6.78 4.06
49.0 Nerolidol trace 1.12 trace 0.02 trace 0.95
* Numbers refer volatile composition relative to the internal standard (ethyl decanoate).

3.6. R e s u l t s of m u l t i v a r i a t e a n a l y s i s on a r o m a c o m p o n e n t s
Figure 1 shows the principal component loading
r e s u l t s of the m u l t i v a r i a t e a n a l y s i s of black tea aroma components. Figure 2
shows the results of the m u l t i v a r i a t e analysis of the aroma components. Each
sample approache green tea with an increase in the second major component
(i.e., positive value) axis 2 and the characteristics of black tea became evident
as the second major component became negative. The samples from Viet Nam
and Turkey were somewhat different in aroma from the Uva and Assam t e a s .
 429

1
0.9
O.S
5«
0.7
0.6
^ 0.5
m

S 0..4
0.3-|
0.2-] 12*

0.1-1
0- 0 . -0.6 -0.4 -0.2 0.2 0.4
axis 2

Figure 1. Scattergram of tea on axis 1 and axis 2 in principal components analysis

l,2,3:green tea, 4,5,6:oolong tea, 7: vie tnam black tea,8:turkey black tea,
9:darjiling tea,10:keemun black tea, ll:assam black tea, 12:sri lanka uva black tea.

axx
5-1

2H
^'
11
• •
— J — — I
-1.5 -1 -0.5 0.5 1-5
axis 2
-1-*

Figure 2. Scattergram of flavor componets on axis 1 and axis 2 in principal components

analysis
430

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