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ABSTRACT
An HPLC method has been used to separate and determine quality-related
chemical components in Zhenong-xiangya and Zhenong-cuiliu, two special
green teas fiom Zhejiang, China. Seventeen or eighteen amino acids and five
catechins were detected. Theamine was the major amino acid by far, its
content reaching 37.7% and 54% of the total amino acids in these teas,
respectively. The catechin content was 154.4 mg g - ' in Zhenong-xiangya
and 170.7 mg g-' in Zhenong-cuiliu. Caffeine contents in the teas studied
were all above 75 mg g-' which was much higher than those previously
reported in green teas. The vitamin C contents were all about 2 mg g - Seven
peaks were resolved in the HPLC projiles of the flavonoid extracts of the two
teas and three of them were identijied as rutin, myricetin and quercetin using
reference compounds.
Key words: Camellia sinensis, special green tea, HPLC, amino acid,
catechin, vitamin C, flavonoid, theamine.
INTRODUCTION
Zhenong-xiangya and Zhenongcuiliu are two special green teas in China which
were first manufactured by the Department of Tea Science at Zhejiang Agricultural
University in 1986 and were awarded prizes for 'Excellent Famous Tea' by the
Zhejiang Tea Science Association in 1987 and 1989. The present research sets out to
investigate the chemical composition of these teas.
54 1
542
10 p1
Amino acid analysis column
62C
Solvent A: 0.2 M sodium citrate buffer, pH 3.0; solvent B:
0.2 M sodium borate buffer, pH 9-6
Pure solvent A to solvent A/B 52:48 v by a linear gradient
during 90 min
0-4 ml min Beckman fluorimetric detector model 157, excitation
338 nm, emission 425 nm filters
0.10 aufs
'
10 pl
p-Bondapak C18, 3.9 x 300 mm
30C
Solvent A: acetic acid/methanol/water (1:1:98 v); solvent
B:
acetic
acid/methanol/dimethylamine/water
(1:1:50:48 v)
Gradient:
Flow rate:
Detector:
Sensitivity:
543
2 p1
p-Bondapak fatty acid column, 4 x 300 mm
550 ml litre-' methanol adjusted to pH 3.0 with H 3 P 0 4
1.2 ml min-'
Ultraviolet detector, 254 nm
0.1 aufs
544
Y R Liang, Z S Liu. Y R
XU.
YL
HU
which 30% was awarded for appearance, 25 % for flavour, 10% for liquor colour,
25% for taste and 10% for infused leaf.
Zhenong-xiangya
Zhenong-cuiliu
Aspartic acid
Threonine
Serine
Glutamic acid
Proline
Glycine
Alanine
Theamine
Valine
Methionine
Isoleucine
Leucine
Tyrosine
Phenylalanine
Histidine
Tryptophan
Lysine
Arginine
Total (%)
1.55
0.984
1.13
5.28
Trace
0.467
0.489
18.97
0.023
Trace
0.106
0.24
0.192
0.271
3.1
0.034
0.026
2.29
3.52
1.59
1.18
1.23
5.09
Trace
0.486
0.684
11.79
0.037
0
0.152
0.244
0.188
0.216
3.65
Trace
0.027
4.48
3.13
545
14
45
(b)
'i
A
I
90
Retention t i m e (rnin)
Fig 1. HPLC analysis of amino acids in two special green teas A, Zhenong-xiangya;B, Zhenongcuiliu. 1 ,
Asp; 2, Thr; 3, Ser; 4, Theamine; 5, Glu; 6, Gly; 7, Ala; 8, Val; 9, Met; 10, Ile; 1 1 , Leu; 12, Tyr; 13, Phe;
14, His; 15, Trp; 16, Lys; 17, Arg.
546
25
0
Retention time (min)
25
Fig 2. HPLC analysis of caffeine and catechins in two special green teas. A. Zhenong-xiangya; B,
Zhenongcuiliu. 1, Caffeine; 2, L-EGC; 3, DL-GC;4, L-EC;5, L-EGCG;6, L-ECG.
TABLE 2
Catechin composition of two special green teas (mg g-')
Sample
Zhenong-xiangya
Zhenongcuiliu
L-EGC
DL-GC
L-EC
L-EGCG
L-ECG
Total
18.42
16.43
069
2.58
10.11
17.13
76.20
75.93
48.99
58-66
154.4
170.7
TABLE 3
Contents of caffeine and vitamin C in two special green teas (mg g-')
Sample
Caffeine
Vitamin C
Zhenong-xiangya
Zhenong-cuiliu
77.2
75-6
2.17
2.08
(=)L
547
23.5
10
TABLE 4
Contents of three identified flavonoids in two special green teas
(mgg-')
Sample
Rutin
Myricetin
Quercetin
Zhenong-xiangya
Zhenongcuiliu
2.53
0.181
1-67
0.404
0
0.075
548
TABLE 5
Mean scores of the two samples evaluated by four senior tea tasters
Sample
Appearance
Flavour
Liquor
colour
Taste
Infused
leaf
Total
Zhenong-xiangya
Zhenongcuiliu
29.3
28.7
24.3
24.5
9.7
9.8
24.5
24.3
9.8
9-8
97.6
96.9
REFERENCES
Nakagawa M 1973 Relationship between chemical composition and taste of green tea. Study
of Tea 40 1-9.
Sakamoto Y 1967 Flavones in green tea, Part I. Isolation and structure of flavones occurring
in green tea infusion. Agric Biol Chem 31 1029-1034.
Sakamoto Y 1969 Flavones in green tea, Part 11. Identification of isovitexin and saponin.
Agric Biol Chem 33 959-961.
Sakamoto Y 1970 Flavones in green tea, Part 111. Structure of pigments IIIa and IIIb. Agric
Biol Chem 34 919-925.
Xiao Weiqiang 1963 About chlorophyll in green tea. Tea Communication (4-5) 71-74.
Yuan Yucheng 1982 Chemical composition of Zhejiang tea. Tea in Zhejiang (4) 27-32.
Yuan Yucheng et a1 1962 Chemical and biochemical studies on tea catechins. Ann Rep Tea
Res Inst Chinese Acad Agric Sci pp 217-222.