International Journal of Food Properties

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Stability of tea polyphenols solution with different

pH at different temperatures

Liang Zeng, Mengjun Ma, Chen Li & Liyong Luo

To cite this article: Liang Zeng, Mengjun Ma, Chen Li & Liyong Luo (2017) Stability of tea
polyphenols solution with different pH at different temperatures, International Journal of Food
Properties, 20:1, 1-18, DOI: 10.1080/10942912.2014.983605

To link to this article: https://doi.org/10.1080/10942912.2014.983605

Published online: 17 Nov 2016.

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INTERNATIONAL JOURNAL OF FOOD PROPERTIES
2017, VOL. 20, NO. 1, 1–18
http://dx.doi.org/10.1080/10942912.2014.983605

Stability of tea polyphenols solution with different pH at different

temperatures
Liang Zenga,b, Mengjun Maa, Chen Lia, and Liyong Luoa,b
a
College of Food Science, Southwest University, Chongqing, China; bTea Research Institute, Southwest University,
Chongqing, China

ABSTRACT ARTICLE HISTORY

Although tea polyphenols possess a variety of biological activities, the oxida- Received 28 May 2014
tive stability of tea polyphenols limits its application in the diet as preventive Accepted 30 October 2014
medicine. To enlarge biological activity of tea polyphenols, we investigated
KEYWORDS
changes of tea polyphenols with different pH (3, 4, 5, 6, and 7) at different
Tea polyphenols; Stability;
temperatures (4, 25, and 100°C). Changes in transmittance, deterioration, color Tea drinks; Temperature; pH
values, and contents of catechins were evaluated. The results showed that tea
polyphenols with a pH of 3–6 remained stable at 4 and 25°C. With increase of
temperature, the tea polyphenol solutions became darker and less green, but
deeper yellow in color. When the heating temperature was 100°C, a significant
reduction in both total catechins and transmittance was observed. Individual
catechins undergo epimerization in this process. Not only temperature and pH,
but also heating time influenced the epimerization. Total contents of catechins
incubated in pH 3, 4, 5, 6, and 7 citrate buffer solutions for 24 h declined by 15,
24, 41, 57, and 96% at 100°C, respectively. Therefore, tea polyphenols were pH-
sensitive: the lower the pH, the more stable the tea polyphenols during
storage. Storing at low temperature and acidic pH conditions did not signifi-
cantly affect the characteristics of tea polyphenols.

Introduction
Tea is one of the most highly consumed beverages in the world.[1–3] Tea polyphenols (TP) are the major
nutraceutical components in tea and account for 30–42% of the dry weight of the solids in brewed green
tea.[4,5] Tea catechins account for 70% of total TP. The major tea catechins are (−)-epigallocatechin-3-
gallate (EGCG), (−)-epicatechin gallate (ECG), (−)-epigallocatechin (EGC), and (−)-epicatechin (EC).
These epicatechins can change to their epimers those are non-epicatechins, i.e., (−)-gallocatechingallate
(GCG), (−)-catechin gallate (CG), (−)-gallocatechin (GC), and (−)-catechin (C). The polyphenolic con-
stituents have drawn wide attention to some extent due to their natural abundance and biological activities,
which include antioxidant,[6,7] anti-mutagenic,[8] anti-cancer,[9] anti-allergic,[10] and anti-ultraviolet (UV)
activities.[11] Therefore, it is important to minimize changes of TP in tea beverages, tea foods, instant tea
powder, and cosmetics during production, distribution, and storage. Figure 1b showed that the transmit-
tance of TP solutions with a pH of 3–6 had almost no significant change and maintained a stable state in 24
h at 25°C. Also, the transmittance of TP solutions had a significant decline at the first hour at 25°C and
declined gradually in the following hours. The transmittance of TP solutions with pH 7 was bigger at 4°C
than that at 25°C after 24 h. Figure 2b showed that the absorbance values of TP solutions with a pH of 3–6
had almost no significant change and maintained a stable state in 24 h at 25°C. Also, the absorbance values
of TP solutions increased gradually with time at 25°C. These results were consistent with the result reported
by Wang, Kim, and Lee.[21] The absorbance of TP solutions had great difference at different pH. The

CONTACT Liang Zeng [email protected] College of Food Science, Southwest University, Tiansheng Road 2, Beibei
Dstrict, Chongqing 400715, China.
© 2017 Taylor & Francis Group, LLC
2 L. ZENG ET AL.

absorbance values of TP solutions with pH 7 was bigger at 25°C than that at 4°C after 24 h, suggesting that
the absorbance values of TP solutions had great difference at different temperatures.
The stability of TP is dependent on both temperature and pH. It was noted that TP degraded
faster with the increase of either pH, oxygen concentration, or temperature.[12] Hong[13] reported
that several factors, including pH, concentration of proteins, antioxidant levels, and the presence of
metal ions, could affect the stability of EGCG, of which pH is probably the most critical. TP solutions
were very unstable in neutral and alkaline solutions and decomposed in a few minutes, whereas, they
were relatively stable under acidic conditions.[14] Tea catechins were found to be unstable in sodium
phosphate buffer (pH 7.4), conditions under which 80% of them were lost in only 3 h;[15] and 40% of
them were lost in 3 h in boiling water.[15] It was also shown that tea catechins degraded by at least
50% during the first month of storage in commercial soft drinks and even in acidic ones.[15] TP
solution degraded irreversibly to a yellowish-brown solution of the deterioration products, mainly
through oxidation and dimer formation.[16] This browning is undesired, e.g., in bottled green tea
beverages and also declines TP biological activities.
To date, in spite of the numerous potential health benefits revealed, TP are scarcely added to food
or drink products, and in the few cases it is the amounts added, and moreover, the residual amounts
left by the time of consumption, are apparently insufficient to produce beneficial effects. It is
important to understand the stability of TP in foods or drinks during processing and storage in
order to gain the optimum health benefits from them. Previous studies evaluated the stability of TP
only from change of the content of catechins, and do not comprehensively demonstrate the changing
trends of TP from sensory evaluation, including transmittance, deterioration, color value, and
nutritional quality. Hence, the objective of this work was to characterize the degradation of TP
under diverse conditions, and to provide improved ways for storing TP and TP products such as tea
beverages, tea foods, instant tea powder, and cosmetics.

Materials and methods

Chemicals
TP (caffeine < 0.5%) were purchased from Changsha Huacheng Biotech, Inc. (Changsha, China), which
were extracted from tea. Acetic acid (high-performance liquid chromatography [HPLC] grade) and
methanol (HPLC grade) were purchased from Chengdu Changzheng Huabo Co., Ltd. (Chengdu,
China). Chemical standards for EGC, EGCG, EC, ECG, C, and GCG were purchased from Sigma
Aldrich Chemical CO., Ltd. (USA). All other chemicals used in this study were analytical grade.

Sample preparation
To examine the stability of TP with different pH (3, 4, 5, 6, and 7) at different temperatures (4, 25,
and 100°C), 50 mg of TP were maintained in 100 mL of 100 mM citrate buffer solutions at different
temperatures for 24 h. The appropriate volume of water was periodically added to compensate for
loss due to evaporation.

Transmittance and deterioration studies by UV spectra measurement

Samples in citrate buffer solutions (pH 3–7) were stored at 4, 25, and 100°C. Aliquots of 2.5 mL of
TP solution were transferred to 1 cm path length spectrophotometer cuvettes and covered with
parafilm to prevent evaporation. The transmittance at 640 nm was observed for 24 h using an
Ultrospec2450 spectrophotometer (Shimadzu Japan).[17,18] The absorbance value at 425 nm was
observed for 24 h using an Ultrospec2450 spectrophotometer (Shimadzu Japan) to study the
deterioration of TP solutions.[16]
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 3

Color of TP in diverse solutions

Samples were placed as a uniform layer (0.5 cm thick) on a 5 cm diameter Petri dish and a
Tristimulus reflectance colorimeter (HunterLab, model D25) calibrated with a white standard tile
(X = 82.45; Y = 84.46; Z = 101.44) were used. Color was recorded using the CIE-L* a* b* uniform
color space (CIE-Lab), where L* indicates lightness, a* indicates hue on a green (–) to red (+) axis,
and b* indicates hue on a blue (–) to yellow (+) axis.[19]

HPLC analysis of tea catechins

The solution filtrated at 0.45 μm was done before injection. Briefly, an HPLC-20 (Shimadzu Japan),
equipped with an auto injector, a C18 reversed-phase column (250 × 4.6 mm /5 um, Thermo) were
used for the analysis of tea catechins. Mobile phases consisted of 2% glacial acetic acid in water
(eluent A) and methanol (eluent B). A gradient system was adopted as follows: 0–25 min, linear
gradient from 18 to 25% B; 25–30 min, linear gradient from 25 to 35% B; 30–32 min, linear gradient
from 35 to 18% B; 32–37 min, 18% B. The sample injection volume was 10 μL. The flow rate was 0.9
mL/min. Tea catechins were detected at 278 nm.

Statistical analysis
All measurements were carried out in triplicate. Data were subjected to one-way analysis of variance
(ANOVA) using the Compare Means Procedure of SPSS Statistics (19.0; SPSS Inc., Chicago, IL). The
least significant difference (LSD) procedure was used to test for differences between means (differences
were considered to be significant when p < 0.05).

Results and discussion

Transmittance change of TP solution with pH at different temperatures
The transmittance at 640 nm can describe the turbidity of the solution.[17] The transmittance of TP
solutions with different pH at different temperatures were monitored for 24 h (Fig. 1a–c).Figure 1a:
Transmittance in the spectra at 640 nm change of TP solutions with a pH of 3–7 at 4°C; B: at 25°C;
and C: at 100°C. Data are expressed as means ± standard deviation (SD) of n = 3 samples. The
different letter in the same temperature indicated that the difference between the treatments is
significantly through LSD test (p < 0.05)
Figure 1a showed that the transmittance of TP solutions with a pH of 3–6 had almost no
significant change and maintained a stable state in 24 h at 4°C. The transmittance of TP solutions
with pH 7 had a significant decline at the first 1 h and maintained a stable state in 10 h, and then had
a significant decline after 24 h.
As shown in Fig. 1c, there were significant differences between pH 3 to pH 7 at 100°C. From
Fig. 1c, we could see that the transmittances of TP solutions were bigger at low pH values. After
boiling for 24 h, the transmittances of TP solutions with a pH of 3–7 were 85.87, 91.37, 84.70, 74.77,
and 50.13, respectively, resulted in about 2, 7, 14, 24, and 49% reduction in transmittance. The
transmittances of TP solutions at the same pH had great differences at the three different tempera-
tures. The higher the temperature and pH was, the greater decline the transmittance. Perhaps
catechins in liquid and solid environments might undergo degradation, oxidation, epimerization,
and polymerization reactions forming new compounds,[20] which resulted in the increase of the
turbidity and the decline of the transmittance of the solution. The higher the temperature was, the
more compounds would be formed.
4 L. ZENG ET AL.

B
100
A a
a a 100 a a a
a a a a
98 a 98 a
Transmittance (%)

Transmittance (%)
b ab b b
96 96
c b bc
94 94 c
92 92 pH = 3
pH = 3 pH = 4 d
90 pH = 4 90
pH = 5 pH = 5
88 pH = 6 88 pH = 6
pH = 7 pH = 7
86 86
e
84 84
0 1 3 5 10 24 0 1 3 5 10 24
Time (h) Time (h)

C
a a a ab ab
100 a a ab
ab b b ab c b
b c
Transmittance (%)

90 bc c
b d
80 c d
pH = 3
70 pH = 4 d e
pH = 5 e
60 pH = 6
pH = 7
50
f
40
0 1 3 5 10 24
Time (h)

Figure 1. Transmittance in the spectra at 640 nm change of TP solutions with pH 3–7 at A: 4°C; B: at 25°C; C: at 100°C. Data are
expressed as means ± SD of n = 3 samples. The different letter in the same temperature indicated that the difference between the
treatments is significantly through LSD test (p < 0.05).

Deterioration of TP solution with different pH at different temperatures

As the appearance of yellow color correlates with oxidation of TP,[13,16] the absorbance values at 425 nm of
solutions can be used to demonstrated the deterioration of TP solution. The absorbance values with
different pH at different temperatures were observed for 24 h (Fig. 2a–c). Figure 2a: Absorbance in the
spectra at 425 nm change of TP with a pH of 3–7 at 4°C; B: at 25°C; and C: at 100°C. Data are expressed as
means ± SD of n = 3 samples. The different letter in the same temperature indicated that the difference
between the treatments is significantly through LSD test (p < 0.05)
Figure 2a showed that the absorbance values of TP solutions with a pH of 3–6 had almost no
significant change and maintained a stable state in 24 h at 4°C. The absorbance values of TP
solutions with pH 7 had a significant increase at the first 1 h and maintained a stable state in 5 h,
and then had a significant increase.
As shown in Fig. 2c, when the temperature was 100°C, the absorbance values had significant
differences between pH 3 to pH 7. From Fig. 2c, we could see that the absorbance values of TP solutions
were bigger at high pH values. The absorbance values increased gradually with time, as expected. There
were great differences in the absorbance values of TP solutions at the same pH at the three temperatures.
The higher the temperature was, the greater increase in the absorbance value.

Color change of TP solution with different pH at different temperatures

Color measurements enabled us to distinguish the influence of the temperature and pH on some
quality characteristics of TP. Changes in color of TP solutions during heating and storage were
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 5

A B
0.25 0.8 pH = 3
pH = 3
pH = 4 pH = 4
0.20 c 0.6 pH = 5 f
pH = 5
Absorbance

Absorbance
pH = 6 pH = 6
b
pH = 7 b b pH = 7 e
0.15 0.4

c d
0.10 0.2 b
a a a a
a a a
a a a a a
0.05 0.0
0 1 3 5 10 24 0 1 3 5 10 24
Time (h) Time (h)

C
1.8 f
pH = 3
1.5 pH = 4
pH = 5 e
Absorbance

1.2 pH = 6
pH = 7 d f
0.9
c e
0.6 b f
d
c e
0.3 bb c d c c
a b ab b
b bcb bc c
0.0
0 1 3 5 10 24
Time (h)

Figure 2. Absorbance in the spectra at 425 nm change of TP with pH 3–7 at A: 4°C; B: at 25°C; C: at 100°C. Data are expressed as
means ± SD of n = 3 samples. The different letter in the same temperature indicated that the difference between the treatments is
significantly through LSD test (p < 0.05).

expressed as L*, a*, and b* values. Table 1a–c showed that the color difference indicator L* values
decreased while indicators a* and b* values increased with the elevation of temperature and pH,
indicating the development of a brown color. The TP solutions, which contained anthocyanins, a*
and b* values were positive, indicating a red and yellow color. The L* value is the indicator of
lightness–darkness, and the higher it is, the lighter is the liquor.[22]
Table 1a showed that the L* and b* values had no significant change at 4°C in 24 h at a pH of 3–6.
When the pH was 7, the L* values decreased and b* values increased with time in 24 h, and resulted
in about 3.74% reduction and 17.6% increase. The a* values had no significant change in 24 h at a
pH of 3–4 and a significant increase at a pH of 5–7.
Table 1b showed that the L* values and b* values had no significant change at 25°C in 24 h at a
pH of 3–6. When the pH was 7, the L* values decreased and b* values increased with time in 24 h,
and resulted in about 13% reduction and 160% increase. L* values of the 25°C samples were
significantly lower than for the 4°C reference ones, indicating a lower lightness during 25°C storage.
The a* values had no significant change in 24 h at a pH of 3–4 and a significant increase at a pH of
5–7. The a* and b* values increased with pH, suggesting that the TP solution became deeper yellow.
There were no statistically significant differences in the three indicators between 4 and 25°C
treatments at a pH of 3–4.
As shown in Table 1c, when the temperature was 100°C, the L* values had a significant decline
after 1 h and declined gradually over time, and the a* and b* values increased with time. However,
there was a large difference in yellowness between 100 and 25°C. Storing at 100°C showed an
increase in b* value from 11 to 47 in 24 h, whereas storing at 25°C increased its b* value from 11 to
only 29. TP solutions stored at 100°C were more prone to color changes. These results suggested that
the TP solution darkened and became deeper yellow as the temperature was raised. What is more,
 6

Table 1a. Color change of TP solution with pH 3–7 at 4°C.

Color change pH 0h 1h 3h 5h 10 h 24 h
L. ZENG ET AL.

L* 3 92.00 ± 1.0 92.13 ± 0.5 92.10 ± 0.4 92.01 ± 0.4 92.31 ± 0.4 91.93 ± 0.2
0a 1a 7a 1a 9a 2a
4 92.00 ± 1.0 91.74 ± 0.4 91.77 ± 0.3 91.80 ± 0.5 92.11 ± 0.7 91.83 ± 0.3
0a 8a 5a 8a 3a 2a
5 92.00 ± 1.0 91.56 ± 0.1 91.67 ± 0.4 91.51 ± 0.5 91.90 ± 0.3 91.49 ± 0.1
0a 7a 5a 1a 3a 2a
6 92.00 ± 1.0 90.98 ± 0.1 91.21 ± 0.3 91.31 ± 0.4 91.50 ± 0.3 91.23 ± 0.1
0a 1b 2ab 3ab 8ab 0ab
7 92.00 ± 1.0 89.18 ± 0.1 88.77 ± 0.7 88.61 ± 0.5 89.02 ± 0.3 88.56 ± 0.5
0a 5b 0b 4b 2b 1b
a* 3 1.73 ± 0.15 1.89 ± 0.14 1.93 ± 0.10 1.95 ± 0.09 1.93 ± 0.16 1.78 ± 0.27
a a a a a a
4 1.73 ± 0.15 2.13 ± 0.40 2.04 ± 0.21 2.10 ± 0.27 2.10 ± 0.31 1.95 ± 0.23
a a a a a a
5 1.73 ± 0.15 2.31 ± 0.09 2.22 ± 0.10 2.18 ± 0.12 2.19 ± 0.09 1.97 ± 0.25
b a ab ab ab b
6 1.73 ± 0.15 2.82 ± 0.16 2.66 ± 0.14 2.53 ± 0.12 2.55 ± 0.08 2.27 ± 0.29
c a a ab ab b
7 1.73 ± 0.15 4.12 ± 0.21 4.16 ± 0.06 4.24 ± 0.05 4.41 ± 0.16 4.11 ± 0.18
c b ab ab a b
b* 3 11.17 ± 1.2 12.24 ± 0.2 12.08 ± 0.1 12.09 ± 0.2 11.91 ± 0.4 11.81 ± 0.2
6b 9a 9ab 9ab 2ab 1ab
4 11.17 ± 1.2 11.73 ± 0.7 11.82 ± 1.0 11.57 ± 0.2 11.26 ± 0.2 11.39 ± 0.0
6a 1a 5a 0a 0a 9a
5 11.17 ± 1.2 11.66 ± 0.7 11.61 ± 0.6 11.82 ± 0.7 11.18 ± 0.7 11.44 ± 0.4
6a 5a 7a 2a 9a 7a
6 11.17 ± 1.2 10.78 ± 0.6 10.78 ± 0.6 10.71 ± 0.2 10.64 ± 0.3 11.02 ± 0.1
6a 5a 5a 7a 2a 0a
7 11.17 ± 1.2 11.19 ± 0.3 11.19 ± 0.3 11.93 ± 0.0 12.81 ± 0.1 13.14 ± 0.3
6b 2b 2b 7b 5ab 9a
Table 1b. Color change of TP solution with pH 3–7 at 25°C.
Color change pH 0h 1h 3h 5h 10 h 24 h
L* 3 92.00 ± 1.0 92.23 ± 0.5 92.19 ± 0.4 92.11 ± 0.5 92.28 ± 0.3 91.88 ± 0.2
0a 2a 6a 4a 5a 8a
4 92.00 ± 1.0 91.91 ± 0.6 91.86 ± 0.5 91.85 ± 0.4 92.12 ± 0.5 91.78 ± 0.3
0a 8a 0a 8a 5a 0a
5 92.00 ± 1.0 91.79 ± 0.3 91.66 ± 0.4 91.77 ± 0.3 91.93 ± 0.2 91.58 ± 0.1
0a 5a 0a 5a 7a 7a
6 92.00 ± 1.0 91.30 ± 0.4 91.33 ± 0.3 91.14 ± 0.2 91.47 ± 0.3 91.14 ± 0.2
0a 0a 9a 3a 6a 5a
7 92.00 ± 1.0 89.16 ± 0.5 88.63 ± 0.6 87.92 ± 0.7 84.41 ± 0.6 79.97 ± 1.2
0a 8b 0b 0b 6c 6d
a* 3 1.73 ± 0.15 1.68 ± 0.12 1.77 ± 0.07 1.88 ± 0.17 1.79 ± 0.10 1.82 ± 0.23
a a a a a a
4 1.73 ± 0.15 1.97 ± 0.38 1.96 ± 0.13 1.88 ± 0.13 1.76 ± 0.10 1.71 ± 0.18
a a a a a a
5 1.73 ± 0.15 2.06 ± 0.05 1.99 ± 0.07 1.95 ± 0.13 1.86 ± 0.16 1.73 ± 0.22
b a a ab ab b
6 1.73 ± 0.15 2.61 ± 0.14 2.42 ± 0.09 2.35 ± 0.09 2.14 ± 0.08 1.98 ± 0.15
d a ab b bc c
7 1.73 ± 0.15 4.51 ± 0.22 4.49 ± 0.15 4.55 ± 0.23 6.00 ± 0.48 8.20 ± 0.87
d c c c b a
b* 3 11.17 ± 1.2 12.10 ± 0.3 12.09 ± 0.1 11.99 ± 0.3 12.17 ± 0.2 12.11 ± 0.5
6a 2a 7a 3a 0a 6a
4 11.17 ± 1.2 11.40 ± 0.2 11.39 ± 0.1 11.61 ± 0.1 11.74 ± 0.3 11.86 ± 0.5
6a 4a 9a 5a 9a 0a
5 11.17 ± 1.2 11.24 ± 0.8 11.33 ± 0.6 11.44 ± 0.7 11.64 ± 0.7 11.92 ± 0.5
6a 2a 3a 6a 2a 7a
6 11.17 ± 1.2 10.43 ± 0.2 10.73 ± 0.1 11.26 ± 0.5 11.74 ± 0.5 12.34 ± 0.7
6ab 7b 7b 5ab 7ab 3a
7 11.17 ± 1.2 11.82 ± 0.3 13.25 ± 0.3 15.19 ± 0.9 21.68 ± 1.4 28.99 ± 2.4
6d 0d 6cd 7c 2b 2a
INTERNATIONAL JOURNAL OF FOOD PROPERTIES
7
 8

Table 1c. Color change of TP solution with pH 3–7 at 100°C.

Color change pH 0h 1h 3h 5h 10 h 24 h
L* 3 92.00 ± 1. 91.23 ± 0.2 90.77 ± 0.2 90.35 ± 0.1 90.43 ± 1.0 88.43 ± 0.1
L. ZENG ET AL.

00a 0a 5b 1b 2b 9c
4 92.00 ± 1. 91.22 ± 0.2 90.61 ± 0.1 90.23 ± 0.2 89.96 ± 0.6 86.56 ± 0.1
00a 5ab 9b 8b 8b 1c
5 92.00 ± 1. 89.56 ± 0.3 88.43 ± 0.6 87.80 ± 0.5 88.19 ± 1.4 84.47 ± 1.5
00a 5b 9b 0b 1b 4c
6 92.00 ± 1. 88.22 ± 0.5 86.71 ± 1.1 85.92 ± 0.7 85.30 ± 2.1 77.48 ± 0.7
00a 3b 4bc 6c 8c 9d
7 92.00 ± 1. 85.30 ± 0.2 81.49 ± 0.4 73.22 ± 1.2 59.54 ± 0.8 43.11 ± 6.5
00a 8b 7b 1c 6d 5e
a* 3 1.73 ± 0.1 1.93 ± 0.19 1.91 ± 0.08 2.18 ± 0.09 2.38 ± 0.32 3.39 ± 0.18
5c c c bc b a
4 1.73 ± 0.1 1.76 ± 0.11 1.59 ± 0.07 1.78 ± 0.07 2.02 ± 0.35 3.82 ± 0.12
5bc bc c bc b a
5 1.73 ± 0.1 2.29 ± 0.14 2.46 ± 0.16 2.80 ± 0.19 2.76 ± 0.64 4.66 ± 0.89
5c bc b b b a
6 1.73 ± 0.1 2.91 ± 0.51 3.54 ± 0.50 4.18 ± 0.32 4.91 ± 1.38 10.18 ± 0.6
5d cd c bc b 1a
7 1.73 ± 0.1 5.34 ± 0.52 10.33 ± 0.3 21.07 ± 1.0 36.40 ± 0.8 37.56 ± 2.2
5d d 2c 5b 8a 7a
b* 3 11.17 ± 1. 14.42 ± 0.1 15.95 ± 0.5 17.83 ± 0.4 19.49 ± 2.1 26.28 ± 0.9
26d 9c 1c 2bc 0b 1a
4 11.17 ± 1. 14.32 ± 0.4 16.28 ± 0.0 17.38 ± 0.4 18.69 ± 1.2 25.57 ± 0.6
26e 9d 8c 7bc 4b 2a
5 11.17 ± 1. 17.77 ± 0.4 19.99 ± 1.0 21.38 ± 1.0 21.20 ± 1.7 26.50 ± 2.7
26d 0c 1bc 2b 4b 1a
6 11.17 ± 1. 19.81 ± 1.0 22.35 ± 1.6 24.19 ± 1.2 25.41 ± 3.6 35.59 ± 1.0
26d 2c 3bc 6b 2b 6a
7 11.17 ± 1. 23.37 ± 0.4 25.36 ± 0.3 31.19 ± 1.5 42.17 ± 1.6 46.83 ± 2.7
26e 7d 8d 5c 8b 7a
Values are expressed as means ± standard deviations of triplicate measurements.
Different superscript lowercase letters (a, b, c, d) in same row indicate that the difference between the treatments is significantly through LSD test (p < 0.05).
T = temperatures.
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 9

the higher the pH was, the greater decline in the L* values. The results suggested that the solution
darkened as the pH raised. These results might be related to the oxidation and degradation of TP
under hot conditions.

Change in contents of catechins of TP solution with different pH at different temperatures

The effect of pH on stability of catechins was best illustrated when the pH value of citrate buffer
solutions were set between 3 and 7. Thermal stability of tea catechins with different pH was
examined at different temperatures (4, 25, and 100°C) for 24 h in the present study (Table 2a–c).
The HPLC analysis showed that the yield of total catechins in TP solution was 388 mg/L (78% of
TP). Among six catechins of TP, EGCG was most abundant (>70% of total catechins), followed by
ECG (>15%), GCG, EGC, EC, and C. Table 2a–c visually illustrated the decreases in the values of
both individual and total catechins levels of the solution at different temperatures.
As shown in Table 2a, total contents of catechins incubated in pH 3, 4, 5, and 6 citrate buffer
solutions for 24 h had almost no significant change and maintained a stable state at 4°C. Catechins
degraded readily at pH 7, 21% of total catechins were lost after 24 h. Moreover, when TP with pH 7
were stored at 4°C for 1 h, a 12% loss of catechins were observed. Storing for an additional 9 h led to
only an additional 3% loss of catechins. Maybe the initial oxygen concentration was higher in the TP
solution so that oxidation of catechins was extensive.[12] Additional storing would decrease the
oxygen concentration in the solution and reduce the reaction between TP and oxygen.
During 4°C storage, all of the six catechins with pH 3–6 had no significant change. When the pH
was 7, the contents of C and EC had no significant change at 4°C. The content of EGC, ECG, GCG,
and EGCG decreased by 32, 9, 12, and 25% in the pH 7 citrate buffer, respectively. EGCG was the
most unstable among all the kinds of catechins when it was dissolved in the pH 7 citrate buffer for 24
h. The ionization state of EGCG, an indication of its proton-donating ability, maybe a factor
contributing to degradation of EGCG.[23]
Table 2b showed that total contents of catechins incubated in pH 3, 4, 5, and 6 citrate buffer
solutions for 24 h had almost no significant change and maintained a stable state at 25°C. When TP
were dissolved in pH 7 buffer, degradation of the catechins was extensive: up to more than 67%. But,
only 21% of total catechins were lost at 4°C after 24 h. So, it had been generally considered that green
tea catechins were stable under low temperature and acidic pH conditions.[24]
During 25°C storage, all of the six catechin with a pH of 3–6 had no significant change. When the
pH was 7, the contents of C had no significant change and EC had a significant decline. The order of
stability of cis-catechins was EC > ECG > EGC > EGCG, and the content decreased by 20, 21, 46, and
83%, respectively, in the pH 7 citrate buffer after 24 h. This was thought to be that EGCG and EGC
have three hydroxyl groups on their B rings, whereas ECG and EC only have two hydroxyl
groups.[21] During the oxidation process, catechins lose one hydrogen radical and form a semiqui-
none radical with an unpaired electron on the oxygen atom. It was suggested that the radical can
form more freely on a ring possessing three hydroxyl groups than on a ring possessing two groups.
This might explain why EGCG and EGC oxidized more rapidly than ECG and EC. No epimerization
had been observed in the samples of strong infusions stored at 5 and 25°C.[25] In neutral pH, EGCG
is easily auto-oxidized. And, the two major oxidative products with dimeric structures formed in a
time-dependent manner and were more hydrophobic or have higher molecular weights.[13] EC and
EGC are the precursors of theaflavin, one of the important oxidation derived components in black
tea. So, the decline of EC and EGC thought to be due to oxidation to form dimers, or the
intermediary products of theaflavin.[26]
As shown in Table 2c, surprisingly, catechins with pH 3 exhibited a remarkable stability at 100°C.
Boiling for 24 h resulted in about 15% reduction in total catechins. Total contents of catechins
incubated in pH 4, 5, 6, and 7 citrate buffer solutions for 24 h declined by 24, 41, 57, and 96%,
respectively. These observations were similar to that previously reported by Chen, Zhu, Tsang, and
Huang,[27] namely, that GTC was relatively stable at pH 3 and 4, but it degraded readily at pH 5 and
 10

Table 2a. Change in contents of catechins of TP solution with pH 3–7 at 4°C.

Compounds pH 0h 1h 3h 5h 10 h 24 h
C 3 3.06 ± 0.58 2.97 ± 0.86 2.57 ± 0.53
2.97 ± 0.55a a a a 2.75 ± 0.51a 2.24 ± 0.66a
4 2.72 ± 0.57 3.16 ± 0.12 3.13 ± 0.18
2.97 ± 0.55a a a a 3.24 ± 0.26a 3.01 ± 0.33a
5 3.39 ± 0.21 3.15 ± 0.22 3.33 ± 0.18
2.97 ± 0.55a a a ab 3.29 ± 0.06ab 3.06 ± 0.07b
L. ZENG ET AL.

6 3.14 ± 0.11 2.96 ± 0.39 3.13 ± 0.11

2.97 ± 0.55a a a a 3.04 ± 0.21a 3.10 ± 0.15a
7 3.14 ± 0.47 3.21 ± 0.52 3.32 ± 0.34
2.97 ± 0.55a a a a 3.15 ± 0.88a 2.99 ± 0.80a
EC 3 10.17 ± 1.04 10.64 ± 0.1 9.93 ± 0.97 10.34 ± 0.7 10.39 ± 0.39
a 7a a 7a 10.39 ± 0.51a a
4 10.17 ± 1.04 9.12 ± 2.04 10.26 ± 0.4 10.12 ± 0.4
a a 4a 6a 9.70 ± 0.65a 9.98 ± 0.92a
5 10.17 ± 1.04 9.90 ± 1.01 9.72 ± 0.93 9.78 ± 0.73
a a a a 8.87 ± 1.36a 8.91 ± 1.19a
6 10.17 ± 1.04 10.71 ± 0.9 10.50 ± 0.9 10.37 ± 0.9 10.39 ± 0.82
a 6a 3a 2a 10.44 ± 0.75a a
7 10.17 ± 1.04 9.74 ± 0.45 9.79 ± 0.46 9.61 ± 0.36
a a a a 9.82 ± 0.65a 9.06 ± 0.64a
EGC 3 17.00 ± 1.00 18.10 ± 0.9 18.20 ± 0.6 17.10 ± 0.7 16.71 ± 1.32
a 6a 0a 2a 17.40 ± 0.69a a
4 17.00 ± 1.00 17.10 ± 0.4 16.83 ± 1.4 16.92 ± 1.4 17.03 ± 0.79
a 5a 6a 1a 17.20 ± 1.88a a
5 17.00 ± 1.00 17.87 ± 0.5 17.58 ± 0.5 18.47 ± 0.3 18.69 ± 1.07
b 9ab 9ab 8a 18.89 ± 0.91a a
6 17.00 ± 1.00 17.31 ± 1.0 17.25 ± 1.8 17.75 ± 0.9 18.10 ± 0.94
a 3a 0a 0a 17.70 ± 0.69a a
7 17.00 ± 1.00 14.19 ± 0.3 14.28 ± 0.4 13.06 ± 0.4 12.43 ± 0.71c 11.56 ± 0.42
a 2b 4b 0c d d
ECG 3 60.07 ± 4.90 59.71 ± 4.7 59.92 ± 4.1 60.62 ± 4.3 61.61 ± 4.75
a 5a 0a 6a 60.50 ± 4.05a a
4 60.07 ± 4.90 58.96 ± 5.0 59.45 ± 4.3 59.80 ± 4.3 61.29 ± 2.59
a 2a 4a 7a 61.48 ± 3.78a a
5 60.07 ± 4.90 61.42 ± 4.4 60.29 ± 4.7 57.90 ± 1.9 59.31 ± 1.73
a 5a 8a 7a 57.84 ± 1.69a a
6 60.07 ± 4.90 57.63 ± 6.3 56.44 ± 3.6 56.37 ± 3.8 59.52 ± 0.94
a 6a 1a 6a 59.25 ± 1.34a a
7 60.07 ± 4.90 53.52 ± 1.3 54.44 ± 1.9 54.22 ± 1.4 54.85 ± 3.34
a 2b 5b 0b 53.44 ± 1.86b b
GCG 3 18.57 ± 1.50 19.37 ± 1.6 18.08 ± 1.2 19.48 ± 1.2 19.65 ± 0.81
(Continued )
Table 2a. (Continued).
Compounds pH 0h 1h 3h 5h 10 h 24 h
a 4a 3a 3a 18.80 ± 1.33a a
4 18.57 ± 1.50 17.85 ± 2.3 20.49 ± 2.1 20.44 ± 1.8 19.42 ± 2.25
a 7a 9a 9a 20.87 ± 1.40a a
5 18.57 ± 1.50 18.98 ± 1.0 18.60 ± 0.9 19.06 ± 1.1 20.21 ± 0.88
a 3a 3a 4a 19.36 ± 1.44a a
6 18.57 ± 1.50 18.62 ± 1.7 18.68 ± 1.9 18.74 ± 1.8 19.82 ± 0.62
a 8a 2a 9a 19.48 ± 0.71a a
7 18.57 ± 1.50 17.25 ± 0.9 17.40 ± 0.7 17.02 ± 0.8 17.65 ± 0.61a 16.27 ± 1.30
a 3ab 1ab 9ab b b
EGCG 3 279.67 ± 10.50a 284.57 ± 7.50a 282.34 ± 9.22a 280.16 ± 7.27a 275.71 ± 6.14a 281.50 ± 2.96a

4 279.67 ± 10. 281.08 ± 4.22a 280.81 ± 4.09a 281.95 ± 3. 286.08 ± 10.57a 274.66 ± 11.63a
50a 43a

6
5 279.67 ± 10.50a 275.20 ± 2.96ab 268.61 ± 3.99b 273.29 ± 2.65ab 276.69 ± 0.92ab 277.27 ± 1.74ab

6 279.67 ± 10.50a 270.58 ± 15.87a 276.17 ± 6.56a 277.28 ± 8.96a 275.84 ± 8.32a 275.96 ± 7.99a
7 279.67 ± 10. 242.69 ± 11 242.45 ± 10 234.30 ± 13 230.32 ± 15.4 210.49 ± 9.8
50a .54b .57b .54b 2bc 4c
Catechins 3 388.43 ± 16.85a 395.45 ± 12.36a 391.43 ± 10.42a 390.27 ± 14.12a 385.54 ± 9.91a 392.10 ± 5.93a

4 388.43 ± 16.85a 386.84 ± 2.44a 391.02 ± 5.51a 392.37 ± 3.88a 398.57 ± 13.49a 385.39 ± 13.24a
388.43 ± 16. 386.77 ± 1. 377.96 ± 8. 381.83 ± 5. 384.93 ± 3.06 387.46 ± 3.3
85a 45a 90a 60a a 8a
5
6 388.43 ± 16. 377.98 ± 25 382.01 ± 14 383.62 ± 16 385.75 ± 10.1 386.89 ± 8.0
85a .51a .78a .55a 9a 0a
7 388.43 ± 16. 340.53 ± 12 341.57 ± 10 331.53 ± 14 326.82 ± 15.9 305.21 ± 9.9
85a .84b .77b .49b 8bc 9c
INTERNATIONAL JOURNAL OF FOOD PROPERTIES
11
 12

Table 2b. Change in contents of catechins of TP solution with pH 3–7 at 25°C.

Compounds pH 0h 1h 3h 5h 10 h 24 h
C 3 2.97 ± 0.55 2.82 ± 0.42 2.67 ± 0.62 2.57 ± 0.47 2.99 ± 1.08
a a a a a 2.66 ± 0.49a
4 2.97 ± 0.55 2.61 ± 0.53 3.13 ± 0.09 3.08 ± 0.17 3.12 ± 0.21
a a a a a 2.51 ± 0.78a
5 2.97 ± 0.55 3.09 ± 0.35 2.81 ± 0.48 3.25 ± 0.15 3.16 ± 0.31
a a a a a 2.98 ± 0.11a
L. ZENG ET AL.

6 2.97 ± 0.55 3.22 ± 0.10 2.69 ± 0.99 3.10 ± 0.29 2.84 ± 0.08
a a a a a 2.77 ± 0.37a
7 2.97 ± 0.55 4.23 ± 0.86 3.83 ± 0.62 4.74 ± 0.27 4.53 ± 0.12
a a a a ab 5.71 ± 0.51a
EC 3 10.17 ± 1.0 10.61 ± 0.0 10.00 ± 0.9 10.28 ± 0.7 9.70 ± 0.50 10.01 ± 0.75
4a 9a 6a 0a a a
4 10.17 ± 1.0 8.84 ± 1.93 10.11 ± 0.4 9.52 ± 0.06 9.17 ± 0.67
4a a 5a a a 9.48 ± 1.34a
5 10.17 ± 1.0 9.80 ± 0.96 9.00 ± 0.12 9.32 ± 0.64 8.32 ± 0.80
4a ab ab ab b 8.42 ± 0.76b
6 10.17 ± 1.0 10.27 ± 1.0 9.85 ± 0.74 9.24 ± 0.92 9.35 ± 0.31
4a 1a a b b 8.35 ± 0.75b
7 10.17 ± 1.0 9.78 ± 0.50 9.22 ± 0.28 8.95 ± 0.28 8.99 ± 0.45
4a ab ab b b 8.08 ± 0.84b
EGC 3 17.00 ± 1.0 17.45 ± 0.7 17.38 ± 0.9 17.09 ± 0.6 17.71 ± 2.0 17.63 ± 1.95
0a 1a 9a 2a 7a a
4 17.00 ± 1.0 16.87 ± 1.2 16.92 ± 1.5 16.83 ± 1.4 16.90 ± 1.6 17.05 ± 1.16
0a 3a 5a 0a 8a a
5 17.00 ± 1.0 17.11 ± 0.4 16.73 ± 1.2 18.27 ± 0.2 18.38 ± 1.2 18.20 ± 1.67
0a 6a 6a 2a 6a a
6 17.00 ± 1.0 16.97 ± 1.0 16.95 ± 1.2 17.43 ± 1.1 16.56 ± 0.0 15.84 ± 2.00
0a 5a 8a 7a 3a a
7 17.00 ± 1.0 14.12 ± 0.8 10.25 ± 0.7 15.63 ± 0.5 13.80 ± 1.5
0a 4b 9c 9ab 1b 9.22 ± 0.87c
ECG 3 60.07 ± 4.9 61.16 ± 4.6 61.07 ± 4.3 60.22 ± 4.3 61.67 ± 4.7 61.00 ± 4.24
0a 2a 7a 0a 0a a
4 60.07 ± 4.9 59.38 ± 4.5 59.36 ± 4.4 59.98 ± 4.2 60.37 ± 3.4 59.34 ± 3.38
0a 8a 4a 3a 5a a
5 60.07 ± 4.9 61.47 ± 5.0 60.00 ± 3.0 58.48 ± 1.3 58.23 ± 1.1 61.00 ± 2.56
0a 3a 9a 4a 2a a
6 60.07 ± 4.9 56.77 ± 3.6 56.20 ± 3.6 56.50 ± 3.0 59.93 ± 3.5 60.57 ± 1.27
0a 0a 1a 9a 2a a
7 60.07 ± 4.9 58.56 ± 4.1 53.20 ± 1.2 52.66 ± 1.8 48.34 ± 2.4 47.49 ± 2.88
0a 4ab 9b 7b 1b c
GCG 3 18.57 ± 1.5 19.32 ± 1.6 17.95 ± 1.1 19.67 ± 1.0 19.19 ± 0.4 19.89 ± 0.69
(Continued )
Table 2b. (Continued).
Compounds pH 0h 1h 3h 5h 10 h 24 h
0a 5a 6a 4a 4a a
4 18.57 ± 1.5 17.41 ± 2.0 20.43 ± 2.2 20.69 ± 1.6 20.18 ± 1.6 20.07 ± 1.57
0ab 7b 4ab 8a 4ab ab
5 18.57 ± 1.5 19.07 ± 1.0 18.73 ± 0.4 19.37 ± 1.0 19.45 ± 1.3 20.49 ± 0.71
0b 7ab 8ab 0ab 2ab a
6 18.57 ± 1.5 20.09 ± 2.2 18.58 ± 1.9 19.55 ± 2.3 20.15 ± 1.9 19.51 ± 0.83
0a 4a 6a 8a 6a a
7 18.57 ± 1.5 17.29 ± 0.6 16.03 ± 0.1 15.63 ± 1.2 14.32 ± 1.2 9.00 ± 0.63c
0a 1ab 1b 6b 3b
EGCG 3 279.67 ± 10.50a 283.32 ± 8.56a 283.05 ± 7.51a 280.60 ± 6.38a 275.65 ± 4.05a 276.89 ± 5.79a
4 279.67 ± 10 278.83 ± 1. 280.24 ± 2. 282.41 ± 2. 283.38 ± 4. 280.17 ± 3.6
.50a 47a 65a 69a 31a 7a

5 279.67 ± 10 282.18 ± 3. 273.47 ± 6. 282.18 ± 7. 282.02 ± 1. 279.58 ± 2.7

.50a 48a 78a 18a 85a 1a

6 279.67 ± 10.50a 267.28 ± 6.83a 268.12 ± 7.33a 266.11 ± 9.31a 266.88 ± 7.23a 266.51 ± 10.87a
7 279.67 ± 10 240.09 ± 10 196.91 ± 13 166.75 ± 15 146.60 ± 10 47.34 ± 2.25
.50a .48b .50c .85d .32e f
INTERNATIONAL JOURNAL OF FOOD PROPERTIES
13
 14

Table 2c. Change in contents of catechins of TP solution with pH 3–7 at 100°C.

Compounds pH 0h 1h 3h 5h 10 h 24 h
C 3 2.97 ± 0.55 2.77 ± 0.12 6.23 ± 0.30 7.14 ± 0.60
c c 3.01 ± 0.24c 5.85 ± 0.60b b a
4 2.97 ± 0.55 2.84 ± 0.14 5.98 ± 1.48 6.71 ± 0.23
c c 4.29 ± 0.43b 5.31 ± 0.50b ab a
5 2.97 ± 0.55 3.31 ± 0.06 6.61 ± 1.12 7.53 ± 0.52
d cd 4.33 ± 0.67c 5.87 ± 0.37b ab a
L. ZENG ET AL.

6 2.97 ± 0.55 5.31 ± 0.53 8.34 ± 1.05 9.44 ± 2.00

c b 7.76 ± 0.32a 8.71 ± 0.94a a a
7 2.97 ± 0.55 10.54 ± 0.5 9.49 ± 2.18a 5.48 ± 0.96 3.43 ± 0.97
b 1a b 12.07 ± 5.32a b b
EC 3 10.17 ± 1.0 11.98 ± 0.8 12.20 ± 0.29 11.46 ± 0.32a 10.18 ± 1.1 9.32 ± 1.52
4b 2a a b 8b b
4 10.17 ± 1.0 9.36 ± 0.12 9.85 ± 0.72 9.24 ± 0.55
4a a 9.45 ± 0.08a 9.90 ± 0.29a a a
5 10.17 ± 1.0 8.67 ± 0.27 8.23 ± 0.48 5.70 ± 1.05
4a b 8.54 ± 0.26b 8.63 ± 0.32b b c
6 10.17 ± 1.0 8.78 ± 0.82 4.51 ± 0.84 5.20 ± 2.45
4a a 6.06 ± 1.36b 5.55 ± 1.05b b b
7 10.17 ± 1.0 5.33 ± 0.52 4.45 ± 0.50b 4.12 ± 0.17 1.81 ± 0.18
4a b c 4.28 ± 0.67bc c d
EGC 3 17.00 ± 1.0 17.63 ± 0.7 16.33 ± 6.60 14.93 ± 2.5 14.46 ± 2.5
0a 5a a 14.69 ± 1.06a 2a 0a
4 17.00 ± 1.0 16.43 ± 1.0 14.06 ± 0.58 14.46 ± 0.30 11.85 ± 0.6 10.75 ± 0.1
0a 1a b b 0c 7c
5 17.00 ± 1.0 14.72 ± 0.4 14.88 ± 0.54 14.67 ± 0.91 10.86 ± 0.9 5.84 ± 0.54
0a 8b b b 8c d
6 17.00 ± 1.0 10.02 ± 1.1 6.10 ± 0.78 5.00 ± 0.76
0a 5b 8.86 ± 0.13b 8.66 ± 0.33b c c
7 17.00 ± 1.0 6.38 ± 0.70 6.05 ± 0.51c 5.21 ± 0.37 2.95 ± 0.49
0a c d 6.99 ± 0.52b d e
ECG 3 60.07 ± 4.9 57.18 ± 5.6 56.61 ± 5.05 54.62 ± 4.56a 51.87 ± 3.4 47.32 ± 1.4
0a 0ab ab b 5b 3b
4 60.07 ± 4.9 59.48 ± 3.4 55.36 ± 0.67 53.56 ± 0.72 53.63 ± 3.4 49.14 ± 2.8
0a 0a ab b 3b 7b
5 60.07 ± 4.9 54.74 ± 2.0 50.11 ± 1.90 41.62 ± 1.6 30.44 ± 2.8
0a 0b bc 48.80 ± 2.38c 5d 0e
6 60.07 ± 4.9 46.25 ± 0.7 27.87 ± 1.24 22.55 ± 1.5 22.45 ± 1.2
0a 8b c 28.23 ± 2.11c 9d 8d
7 60.07 ± 4.9 24.13 ± 1.9 18.63 ± 2.87 8.62 ± 0.14 2.35 ± 1.44
0a 2b c 16.57 ± 2.77c d e
GCG 3 18.57 ± 1.5 17.54 ± 1.2 20.25 ± 1.75 20.52 ± 2.60 23.16 ± 1.2 29.99 ± 2.0
(Continued )
Table 2c. (Continued).
Compounds pH 0h 1h 3h 5h 10 h 24 h
0c 1c bc bc 7b 1a
4 18.57 ± 1.5 20.95 ± 1.0 24.65 ± 0.93 29.78 ± 1.06 31.13 ± 0.4 37.36 ± 1.7
0e 9d c b 8b 7a
5 18.57 ± 1.5 32.80 ± 2.1 62.92 ± 3.95 71.46 ± 19.0 106.80 ± 8. 84.54 ± 2.3
0d 5d c 7bc 23a 3b
6 18.57 ± 1.5 80.57 ± 5.0 130.02 ± 3.2 128.32 ± 6.3 115.08 ± 3. 64.15 ± 4.3
0e 8c 7a 2a 02b 9d
7 18.57 ± 1.5 99.16 ± 0.9 51.75 ± 2.34 36.66 ± 1.75c 4.21 ± 0.13
0d 1a b e 0.46 ± 0.10f
EGCG 3 279.67 ± 10.50a 258.42 ± 6.81b 253.93 ± 4.98bc 241.35 ± 11.64c 234.89 ± 6.57c 220.07 ± 5.91d
4 279.67 ± 10 275.76 ± 6. 254.69 ± 6.4 252.71 ± 7.7 231.87 ± 10 181.99 ± 9.
.50a 30a 0b 5b .96c 46d

5 279.67 ± 10 251.99 ± 2. 212.43 ± 1.6 183.97 ± 3.9 147.68 ± 8. 94.91 ± 9.0

.50a 27b 3c 0d 10e 4f
6
279.67 ± 10.50a 186.71 ± 8.11b 117.63 ± 9.57c 109.81 ± 7.07c 91.94 ± 5.44d 59.19 ± 4.85e
7 279.67 ± 10 81.64 ± 2.6 55.72 ± 4.39 40.22 ± 3.34 8.71 ± 0.94 4.57 ± 0.70
.50a 0b c d e e
Catechin s 3 388.43 ± 16 365.52 ± 14 362.33 ± 13. 348.49 ± 14. 341.26 ± 1. 328.30 ± 8.
.85a .34b 86bc 32bc 00c 78c
388.43 ± 16.85a 384.81 ± 8.96ab 362.50 ± 6.47bc 365.71 ± 6.92b 344.30 ± 12.90c 295.20 ± 9.45d
4
5 388.43 ± 16 366.23 ± 4. 353.21 ± 5.7 333.40 ± 16. 321.79 ± 2. 228.96 ± 10
.85a 74b 5b 59c 06c .75d
6 388.43 ± 16 337.64 ± 12 298.19 ± 7.6 289.28 ± 10. 248.51 ± 7. 165.42 ± 9.
.85a .06b 2c 43c 38d 82e
7 388.43 ± 16 227.18 ± 5. 146.08 ± 6.3 116.80 ± 4.7 36.36 ± 2.3 15.58 ± 1.6
.85a 68b 2c 9d 6e 6f
Values are expressed as means ± standard deviations of triplicate measurements.
Different superscript lowercase letters (a, b, c, d) in same row indicate that the difference between the treatments is significantly through LSD test (p < 0.05).
INTERNATIONAL JOURNAL OF FOOD PROPERTIES
15
16 L. ZENG ET AL.

6. The higher the pH value of the medium, the greater the percentage of catechins that degraded.
Furthermore, there was a declining trend in total catechins with increase of temperature, which
suggested that some of the catechins were oxidized (besides the epimerization) and they might be
responsible for the changes in the liquor color.[22]
The four cis-configured catechins (EGCG, EGC, ECG, and EC) degraded steadily over time,
whereas the two trans-configured catechins (GCG and C) increased during the first heating time and
then decreased with time. These results suggested that epimerizations of catechins took place under
the heating conditions. Courbat[28] pointed out that a rapid epimerization of catechins occurred in
alkaline solution. Nakagawa[29] reported that the isomerization of catechins in the roasting process of
green tea was not racemization but epimerization. In addition to oxidation products, EGCG could be
degraded to GCG or EGCG dimers.[30] We observed increase in GCG concurrent with EGCG
reduction at the first 1 h. Therefore, it was concluded that the high temperature led to the
epimerization. It had been recognized that catechins undergo epimerization at the C-2-position in
hot aqueous solution and this epimerization could change the epi-structured catechins to non-epi-
structured catechins.[20] The relatively high amount of GCG found in some tea drinks was most
likely the epimerization of EGCG during autoclaving.[27] We also observed increase in C concurrent
with EC. The degradation of EC and EGCG was obviously dependent on the pH of the solution. The
reaction scarcely proceeded in slightly acid media at pH < 5; however, it was accelerated at pH > 6.
So, production of trans-catechins (GCG, C) increased with increasing pH levels, suggesting that
isomerization was more favored with increases in pH levels. Inferred from their stereochemical
configuration, catechins with their “2,3-trans” structure are thermodynamically more stable than
epicatechins, which are “2,3-cis,” so catechins isomerized at a lower rate than epicatechins.[21]
Increased reaction products, GCG and C, finally decreased with prolonged heating process, which
indicated that after the catechins reached the maximum level of epimerization, the predominant
change become the degradation or oxidation of the catechins.[26] The decline did not happen at a pH
of 3–4 in 24 h. And, it happened after 1, 3, and 10 h at a pH of 7, 6 and 5, respectively. Furthermore,
it was thought that not only temperature and pH, but also heating time influenced the epimerization.
These results were similar to that previously reported by Komatsu,[25] namely, that a dominant
reaction of EC in slightly acidic media was isomerization as far as the reaction fitted an apparent first
order kinetics, and once other reactions, such as oxidation and/or polymerization, proceeded, the
first order kinetics was disturbed. In this process, there are several competing reactions occurring,
including epimerization, oxidation, and degradation making prediction of the reaction of catechins
more complex.

Conclusion
The results of this study highlighted the importance of temperature and pH in impacting the stability
of TP. According to the stability testing, the present results suggested that TP were stable under low
temperature and acidic pH conditions. The stability of TP were pH-dependent: the higher the pH,
the more unstable the TP. Transmittance, deterioration, color, and contents of catechins were
significantly affected at a pH of 7 or 100°C. These results were the initial step in understanding
the behavior of TP in liquid systems, and the stability of TP should also be studied in solid systems,
which could help us use TP in foods or drinks optimally. The results of the present study for the
stability of TP suggest that the optimal pH tea drink should be below pH 7 and stored at 4 or 25°C
against browning. We can protect TP against degradation and oxidization by adjusting pH and being
stored at low temperature when tea drinking was obtained. Moreover, our laboratory will establish
models of TP-caffeine, TP-tea protein, caffeine-protein, and TP-caffeine-protein, and comprehen-
sively analyze the formation mechanism of green tea cream.
 INTERNATIONAL JOURNAL OF FOOD PROPERTIES 17

Funding
This work was supported by the National Natural Science Foundation of China (Grant no. 31100500), the
Fundamental Research Funds for the Central Universities (XDJK2013B036).

References
1. Razic, S.; Kuntic, V. Diverse Elements in Herbal Tea Products Consumed in Serbia Using Inductively Coupled
Plasma Mass Spectrometry. International Journal of Food Properties 2013, 16, 1–8.
2. Altunkaya, A. Partial Purification and Characterization of Polyphenoloxidase from Turkish Tea Leaf (Camellia
Sinensis L.). International Journal of Food Properties 2014, 17, 1490–1497.
3. Singh Arora, D.; Jeet Kaur, G.; Kaur, H. Antibacterial Activity of Tea and Coffee: Their Extracts and
Preparations. International Journal of Food Properties 2009, 12, 286–294.
4. Khan, N.; Mukhtar, H. Tea Polyphenols for Health Promotion. Life Sciences 2007, 81, 519–533.
5. Yang, C.S.; Lambert, J.D.; Sang, S. Antioxidative and Anti-Carcinogenic Activities of Tea Polyphenols. Archives
of Toxicology 2009, 83, 11–21.
6. Pekal, A.; Drozdz, P.; Pyrzynska, K. Comparison of the Antioxidant Properties of Commonly Consumed
Commercial Teas. International Journal of Food Properties 2012, 15, 1101–1109.
7. Lin, C.C.; Li, C.W.; Shih, Y.T.; Chuang, L.T. Antioxidant and Anti-Inflammatory Properties of Lower-
Polymerized Polyphenols in Oolong Tea. International Journal of Food Properties 2014, 17, 752–764.
8. Zhao, X.; Wang, Q.; Li, G.; Chen, F.; Qian, Y.; Wang, R. In Vivo Antioxidant, Anti-Mutagenic, Anti-Cancer and
Anti-Angiogenic Effects of Chinese Bowl Tea. Journal of Functional Foods 2014, 7, 590–598.
9. Butt, M.S.; Ahmad, R.S.; Sultan, M.T.; Nasir Qayyum, M.M.; Naz, A. Green Tea and Anticancer Perspectives:
Updates from Last Decade. Critical Reviews in Food Science and Nutrition 2015, 55, 792–805.
10. Maeda-Yamamoto, M.; Tachibana, H. Anti-Allergic Action of 0-Methylated EGCG in Green Tea Cultivar
Benifuuki. Journal of Food and Drug Analysis 2012, 20, 313–317.
11. Matsuta, T.; Sakagami, H.; Kitajima, M.; Oizumi, H.; Oizumi, T. Anti-UV Activity of Alkaline Extract of the
Leaves of Sasa Senanensis Rehder. In Vivo 2011, 25, 751–755.
12. Zimeri, J.; Tong, C. Degradation Kinetics of (−)-Epigallocatechin Gallate as a Function of pH and Dissolved
Oxygen in a Liquid Model System. Journal of Food Science 1999, 64, 753–758.
13. Hong, J.; Lu, H.; Meng, X.; Ryu, J.H.; Hara, Y.; Yang, C.S. Stability, Cellular Uptake, Biotransformation, and
Efflux of Tea Polyphenol (−)-Epigallocatechin-3-Gallate in HT-29 Human Colon Adenocarcinoma Cells.
Cancer Research 2002, 62, 7241–7246.
14. Zhu, Q.Y.; Zhang, A.; Tsang, D.; Huang, Y.; Chen, Z.Y. Stability of Green Tea Catechins. Journal of Agricultural
and Food Chemistry 1997, 45, 4624–4628.
15. Lun Su, Y.; Leung, L.K.; Huang, Y.; Chen, Z.Y. Stability of Tea Theaflavins and Catechins. Food Chemistry
2003, 83, 189–195.
16. Shpigelman, A.; Israeli, G.; Livney, Y.D. Thermally-Induced Protein-Polyphenol Co-Assemblies: Beta
Lactoglobulin-Based Nanocomplexes as Protective Nanovehicles for EGCG. Food Hydrocolloids 2010, 24,
735–743.
17. Yan, S.H.; Jin, D. Agglomeration and Re-Dissolution of Polyphenol-Alkaloids in Tea Extract. Tea 1981, 2, 27–
30 (in Chinese).
18. Chen, J.; Liu, Z.H.; Yang, D.X.; Guo, X.C.; Li, H.; Ni, X.W. Low-Temperature Extraction of Green Tea and Its
Effect on Cream Down. Food Science 2012, 33, 47–51 (in Chinese).
19. Larrauri, J.A.; Rupérez, P.; Saura-Calixto, F. Effect of Drying Temperature on the Stability of Polyphenols and
Antioxidant Activity of Red Grape Pomace Peels. Journal of Agricultural and Food Chemistry 1997, 45, 1390–
1393.
20. Ananingsih, V.K.; Sharma, A.; Zhou, W. Green Tea Catechins During Food Processing and Storage: A Review
on Stability and Detection. Food Research International 2013, 50, 469–479.
21. Wang, L.F.; Kim, D.M.; Lee, C.Y. Effects of Heat Processing and Storage on Flavanols and Sensory Qualities of
Green Tea Beverage. Journal of Agricultural and Food Chemistry 2000, 48, 4227–4232.
22. Kim, E.; Liang, Y.; Jin, J.; Sun, Q.; Lu, J.; Du, Y.; Lin, C. Impact of Heating on Chemical Compositions of Green
Tea Liquor. Food Chemistry 2007, 103, 1263–1267.
23. Proniuk, S.; Liederer, B.M.; Blanchard, J. Preformulation Study of Epigallocatechin Gallate, a Promising
Antioxidant for Topical Skin Cancer Prevention. Journal of Pharmaceutical Sciences 2002, 91, 111–116.
24. Roginsky, V.; Alegria, A.E. Oxidation of Tea Extracts and Tea Catechins by Molecular Oxygen. Journal of
Agricultural and Food Chemistry 2005, 53, 4529–4535.
25. Komatsu, Y.; Suematsu, S.; Hisanobu, Y.; Saigo, H.; Matsuda, R.; Hara, K. Effects of pH and Temperature on
Reaction Kinetics of Catechins in Green Tea Infusion. Biosci Biotech Biochem 1993, 57, 907–910.
26. Wang, H.; Helliwell, K. Epimerisation of Catechins in Green Tea Infusions. Food Chemistry 2000, 70, 337–344.
18 L. ZENG ET AL.

27. Chen, Z.Y.; Zhu, Q.Y.; Tsang, D.; Huang, Y. Degradation of Green Tea Catechins in Tea Drinks. Journal of
Agricultural and Food Chemistry 2001, 49, 477–482.
28. Courbat, P.; Weith, A.; Albert, A.; Pelter, A. Contribution to Study of Behavior of Catechin in Alkaline-
Medium. Helvetica Chimica Acta 1977, 60, 1665–1675.
29. Nakagawa, M. The Nature and the Origin of Polyphenols in Hoji-Cha (Roasted Green Tea). Agricultural and
Biological Chemistry 1967, 31, 1283–1287.
30. Sang, S.; Lee, M.J.; Hou, Z.; Ho, C.T.; Yang, C.S. Stability of Tea Polyphenol (-)-Epigallocatechin-3-Gallate and
Formation of Dimers and Epimers Under Common Experimental Conditions. Journal of Agricultural and Food
Chemistry 2005, 53, 9478–9484.