Food Bioprocess Technol (2013) 6:2177–2181

DOI 10.1007/s11947-011-0739-8

COMMUNICATION

Inhibitory Effects of Green Tea Polyphenols

on the Retrogradation of Starches from Different Botanical
Sources
Huaxi Xiao & Qinlu Lin & Gao-Qiang Liu & Yue Wu &
Wei Wu & Xiangjin Fu

Received: 24 July 2011 / Accepted: 14 November 2011 / Published online: 4 December 2011
# Springer Science+Business Media, LLC 2011

Abstract This paper aims at investigating the effects of green rice and maize starch gels with 10% and 15% GTPs had
tea polyphenols (GTPs) on the retrogradation of potato, rice, almost no recrystallization of the retrogradation. It is
and maize starches. Starch/GTPs mixing ratios were 2/0, 2/ concluded that the addition of GTPs considerably inhibits
0.1, 2/0.2, and 2/0.3 (w/w) that equated with starch containing the retrogradation of rice, maize, and potato starches.
0%, 5%, 10%, and 15% GTPs (based on starch weight),
respectively. The analytic samples had a water–starch/GTPs Keywords Green tea polyphenols . Rice starch . Maize
ratio of 2:1. The effects of GTPs on the retrogradation of starch . Potato starch . Retrogradation . Differential scanning
these starches were analyzed by differential scanning calorimetry . X-ray diffraction
calorimetry (DSC) and X-ray diffraction (XRD). The
analysis of DSC indicates that the temperature and enthalpy
of gelatinization of all the tested starches decreased as the Introduction
GTPs level increased. After storage at 4 °C, retrogradation
enthalpy (ΔHret) for potato starches with 5%, 10%, and 15% Starches from various plant sources, such as corn, potato,
GTPs appeared after 10 days of storage, however ΔHret for and rice, have received extensive attention in relation to
rice and maize starches with 10% and 15% GTPs did not structural and physicochemical properties (Kuakpetoon and
appear until storage of 20 days, and both the ΔHret and Wang 2001; Yu et al. 2010; Li et al. 2011; Xiao et al. 2011).
degree of retrogradation of the three starches significantly Potato starch (PS) has a higher phosphate monoester
decreased with the increased level of GTPs. After 10 days of content, however rice and maize starches (RS, MS) are
storage at 4 °C, the analysis of XRD shows that the intensity rich in free fatty acids content which result in higher
of X-ray diffraction peaks of potato starch gradually transition temperatures due to amylose–lipid complex
decreased with the concentration of GTPs increasing, and formation. The uses of starches are limited due to their
high retrogradation tendency during storage that shortens
H. Xiao : Q. Lin (*) : G.-Q. Liu (*) : Y. Wu : W. Wu : X. Fu their shelf lives.
National Engineering Laboratory for Rice Industrially, starch is subjected to various chemical or
and By-products Further Processing,
Central South University of Forestry & Technology,
physical modifications to prevent starch retrogradation.
Changsha 410004, People’s Republic of China Retrogradation of starch has been reported to be sup-
e-mail: gaoliuedu@yahoo.com.cn pressed by cross-linking using sodium trimetaphosphate
and sodium tripolyphosphate mixture (Chung et al.
H. Xiao : Q. Lin : Y. Wu : W. Wu : X. Fu
Department of Food Science and Technology,
2004). Both normal and waxy oxidized starches showed
Central South University of Forestry & Technology, lower values of retrogradation compared to their native
Changsha 410004, People’s Republic of China counterparts (Kuakpetoon and Wang 2001). However,
many chemical modification used in food products are
G.-Q. Liu
College of Life Science and Technology,
not enough safe for health due to questionable chemicals.
Central South University of Forestry & Technology, In addition, physical modification can prevent the retro-
Changsha 410004, People’s Republic of China gradation of rice flour but not rice starch (Wu et al. 2010).
2178 Food Bioprocess Technol (2013) 6:2177–2181

Tea extracts have drawn increased attention in recent tography with a DAD detector using a C18 reversed phase
years, not only as flavoring and food preservatives due to column (250×4.6 mm/5 μm, Waters). Mobile phases
their antioxidant activity and antimicrobial effect but also consisted of 0.1% methanoic acid in methanol (eluent A)
for preventing firmness of foods. Zhou et al. (2008) and methanoic acid in water (eluent B). A gradient system
reported that polysaccharides of tea extract could retard was adopted from Wang and Zhou (2004) with a slight
the retrogradation of wheat starch. In green tea the main modification: 0–10 min, 10% B; 10–30 min, linear gradient
compounds are polyphenols. This group of polyphenols is from 10% to 30% B; 30–36 min, linear gradient from 30%
called tea catechins (Wu et al. 2009). to 45% B; 36–46 min, linear gradient from 45% to 60% B;
Previous studies have mostly focused on excellent antibac- 46–50 min, 60% B; and 50–55 min, linear gradient from
terial and antioxidative activities of polyphenols (Zduńczyk et 60% to 10% B. Postrun time was 5 min. Standard curves of
al. 2002; Feillet-Coudray et al. 2009; Périno-Issartier et al. peak area, as a function of the concentrations of EGCG,
2011). There are a few studies on the relationship between ECG, EGC, and EC, were prepared for quantitative
starch and tea polyphenols (TPs). Zhu et al. (2009) studies analysis.
the effects of extract of phytochemicals from green tea on the
pasting, thermal, and gelling properties of wheat starch. Our Differential Scanning Calorimetry
previous study investigated the effect of TPs on the
retrogradation of a high-amylose rice starch (Wu et al. Gelatinization and retrogradation thermal properties of the
2009). However, the interaction and association between TPs samples were measured using 61 DSC-Pyris Diamond
and retrogradation of rice starch, maize, and potato starches (Perkin-Elmer Corp., Norwalk, CT, USA). Powdered starch
are not well understood. In this work, green tea polyphenols and GTPs were mixed at the ratios of starch/GTP for 2/0, 2/
(GTPs) were used and their effects on the retrogradation of 0.1, 2/0.2, and 2/0.3 (w/w) that equated with starch
starches from different botanical sources were investigated containing 0%, 5%, 10%, and 15% GTPs (based on starch
using differential scanning calorimetry (DSC) and X-ray weight), respectively. The calorimeter was calibrated with
diffraction (XRD). an indium standard. Samples of the mixtures of starch and
GTPs (about 2 mg) were accurately weighed into aluminum
DSC pans, and deionized water was added by micropipette
Materials and Methods at a water sample ratio of 2:1. The sample pans were sealed
and equilibrated at room temperature for 24 h before
Materials analysis. For all DSC runs, a sealed empty aluminum pan
was used as reference. The samples were heated. The
RS, MS, and PS were purchased from Puer Biological scanning temperature range and the heating rate were 25–
Technology Co. Ltd. (Yun Nan, China). Powdered GTPs 100 and 10 °C/min, respectively. The onset temperature To,
were obtained from Novanat Bioresources Co. Ltd. peak temperature Tp, and conclusion temperature Tc were
(Shanghai, China) with total polyphenols of 99.97%. determined from the first run heating DSC curves, the range
Tea catechins, (−)-epigallocatechin gallate (EGCG), (−)- of gelatinization temperature (R=Tc–To) was calculated.
epigallocatechin (EGC), (−)-epicatechin gallate (ECG), Enthalpy of gelatinization (ΔHgel) was evaluated based on
and (−)-epicatechin (EC) were purchased from Sigma- the area of the main endothermic peak. Then the gelatinized
Aldrich®, USA. samples were stored at 4 °C for 5, 10, and 20 days and
rescanned under the same conditions to determine the
Methods temperature and enthalpy changes due to retrogradation.
The retrogradation temperature (To, Tp, and Tc) and
Determination of Amylose and Moisture Contents enthalpy (ΔHret) were determined from the second run
heating. In addition, the degree of retrogradation (DR%)
The amylose contents of rice, maize, and potato starches was calculated as the ratio of the retrogradation enthalpy to
were determined using the colorimetric method described the gelatinization enthalpy in run heating (Rodríguez-
by McGrance et al. (1998). The moisture contents of these Sandovala et al. 2008). Analyses were performed in
starch samples were determined by heating at 105 °C for triplicate.
5 h.
X-ray Diffraction
Analysis of Polyphenols of GTPs
The samples as same as DSC analysis were rice, maize, and
The contents of polyphenols, EGCG, ECG, EGC, and EC potato starches containing 0%, 5%, 10%, and 15% GTPs
of GTPs were analyzed by high-pressure liquid chroma- (based on starch weight). Double deionized water was
Food Bioprocess Technol (2013) 6:2177–2181 2179

added to the samples and gelatinized by steam heating for sample contained of 46.8% EGCG, 24% ECG, 12.8%
20 min in a closed thermostatic water bath. These samples EGC, and 8% EC.
were cooled to room temperature then stored at 4 °C for
10 days. The freeze-dried samples were ground and powder Thermal Properties
passed through a 100-mesh sieve before testing. The
recrystallization analysis of samples was carried out using The gelatinization transition temperatures and gelatinization
a Rigaku D-Max-2500 X-ray diffractometer (Rigaku, enthalpy of samples with added GTPs at various ratios are
Tokyo, Japan) equipped with a copper tube operating at shown in Table 1. The gelatinization temperatures (To, Tp,
40 kV and 250 mA. Diffractograms were obtained, the and Tc) and the enthalpy value of gelatinization (ΔHgel)
scanning region of the diffraction angle (2θ) was from 4° to significantly decreased with the content of the GTPs
50° at a rate of 4°/min, a step size of 0.02°, 1°/1° increasing. The range of gelatinization (R) of rice and
divergence slit/scattering slit, and 0.30-mm receiving slit. maize starches significantly increased as the GTPs level
MDI Jade 5.0 was used to analyze the diffractograms. increased. The increase in R implied that after adding GTPs
to starches, the crystallinity of starches became heteroge-
Statistical Analysis neous. The effect of GTPs on the gelatinization enthalpy
was to disrupt the crystallites. It may be that the hydrophilic
The data reported in the tables were average of triplicate character of GTPs OH groups interacted with side chains of
observations. Data obtained were analyzed by analysis of amylopectin and bind to the amorphous region of starch
variance using SPSS for windows version 13.0 following granules to various degrees and thus change the coupling
the procedure described by Miller and Miller (1993). forces between the crystallites and the amorphous matrix
Confidence interval of sample means was reported at the (Wu et al. 2009). This results in an easy hydration of starch
95% confidence probability. Comparisons of means were granules, requiring less thermal energy for gelatinization
made using least significant difference (LSD) and shortest (Zhu et al. 2009).
significant ranges at 5% significance level (p<0.05). The retrogradation enthalpy and retrogradation ratio of
gelatinized starch with added GTPs at various ratios are
shown in Table 2. Native maize starch shows the highest
Results and Discussion ΔHret. The ΔHret and DR% of gelatinized starches
significantly increased with storage time increasing. How-
Chemical Compositions of Starch and GTPs Samples ever, when GTPs was added to rice, maize, and potato
starches, ΔHret of all samples with GTPs could not be
The amylose contents in rice, maize, and potato starches detected after 5 days of storage. The ΔHret of rice and
were 28.3%, 27.6%, and 20.3%, respectively, while the maize starches with only 5% GTPs were detected after
moisture contents were 11.2%, 11.6%, and 10.8%, respec- 10 days of storage and were 2.33 and 3.13 J/g, respectively.
tively. All starch samples contained crude protein less than However, the ΔHret of potato starches with 5%, 10%, and
0.44% and ash less than 0.15% (data from Puer Co.). GTPs 15% GTPs were detected after 10 days of storage. Only

Table 1 Gelatinization thermal

properties of the mixtures of Samples To (°C) Tp (°C) Tc (°C) ΔHgel (J/g) R (°C)
starches from different botanical
sources and green tea polyphe- RS + 0% GTP 66.39±0.48a 71.40±0.57a 76.12±0.42a 9.68±0.21e 9.73h
nols at various ratios RS + 5% GTP 63.56±0.35b 69.78±0.39b 75.46±0.21b 9.66±0.17e 11.90d
RS + 10% GTP 61.33±0.43c 67.39±0.23c 73.31±0.38d 5.86±0.12h 11.98d
RS + 15% GTP 59.26±037d 66.04±0.19d 71.73±0.22e 3.25±0.10j 12.47a
The values are average ± stan-
dard deviation, n=3. Compari- MS + 0% GTP 66.55±0.51a 71.16±0.68a 75.02±0.49c 10.62±0.22b 8.47i
sons of the values were made MS + 5% GTP 62.98±0.41b 68.94±0.45b 73.23±0.39d 9.04±0.21f 10.25g
using LSD, values followed by MS + 10% GTP 59.48±024d 66.37±051d 71.19±0.28f 6.23±0.13g 11.71e
the same letter in the same
MS + 15% GTP 57.81±0.23e 64.66±034e 69.97±0.15g 4.18±0.09i 12.16b
column are not significantly
different (p<0.05) PS + 0% GTP 57.52±0.19e 62.40±026f 69.03±0.21h 12.16±0.31a 11.51f
R the range of gelatinization PS + 5% GTP 55.32±0.28f 60.73±0.35g 67.05±0.12i 10.41±0.21c 11.73e
(Tc–To), RS rice starch, MS PS + 10% GTP 52.68±0.31g 58.00±0.47h 64.66±0.23j 10.17±0.32d 11.98d
maize starch, PS potato starch, PS + 15% GTP 51.78±0.25h 57.16±0.36i 63.79±0.16k 9.84±0.18e 12.01c
GTP green tea polyphenols
2180 Food Bioprocess Technol (2013) 6:2177–2181

Table 2 The enthalpy and de-

gree of retrogradation of mix- Samples 5 days 10 days 20 days
tures of starches from different
botanical sources and green tea ΔHret (J/g) DR% ΔHret (J/g) DR% ΔHret (J/g) DR%
polyphenols (GTP) at various
ratios after 4 °C storage RS + 0% GTP 2.18±0.11c 22.5b 4.98±0.39b 51.4b 7.65±0.52b 79.0a
RS + 5% GTP NDa –b 2.33±019f 24.1d 4.43±0.28 45.9f
RS + 10% GTP ND – ND – 1.77±0.12i 30.2g
RS + 15% GTP ND – ND – 0.38±0.08 11.7i
The values are average ± stan- MS + 0% GTP 3.14±0.21a 29.6a 5.68±0.43a 53.5a 8.01±0.58a 75.4b
dard deviation, n=3. Values fol-
MS + 5% GTP ND – 3.13±0.26e 34.6c 5.52±0.39f 61.1c
lowed by the same letter in the
same column are not signifi- MS + 10% GTP ND – ND – 3.49±0.29 56.1e
cantly different (p<0.05) MS + 15% GTP ND – ND – 1.14±0.10j 27.3h
RS rice starch, MS maize starch, PS + 0% GTP 2.68±0.14b 22.0b 4.13±0.38c 34.0c 7.48±0.49c 61.5c
PS potato starch, GTP green tea PS + 5% GTP ND – 3.52±0.27d 33.8c 6.08±0.42d 58.4d
polyphenols
a
PS + 10% GTP ND – 2.13±0.22f 20.9e 5.71±0.37e 56.1e
Not detectable
b
PS + 15% GTP ND – 1.01±0.11g 10.3f 4.43±031g 45.0f
Not calculated

after 20 days of storage, the rice and maize starches with scattering angle, at which the diffraction intensities ob-
10% and 15% GTPs began to show retrogradation served, was 2θ. The diffraction pattern of retrograded starch
endotherm on the DSC. Twenty days after the DSC test, it is clearly distinct from that of native starch. In native rice,
was also clearly shown that ΔHret and DR% significantly maize, and potato starches, rice and maize starches showed
decreased with the concentration of GTPs increasing, but it typical A-type XRD patterns with strong peaks at 2θ close
was observed that the decreases in ΔHret and DR% of to 15.4, 17.3, 18.5, and 23.5, and potato starch showed
potato starch were lower compared to those of rice and typical B-type XRD patterns. The retrograded starch (B-
maize starches. These results suggest that the GTPs could type crystal) is characterized by a well-defined peak at 17°
significantly inhibit the retrogradation of rice, maize, and (2θ, Kim et al. 1997).
potato starches, and the inhibitory effects for rice and maize Figure 1 shows that retrograded potato starches with 0%,
starches were more marked than that for potato starch. 5%, 10%, and 15% GTPs gave a B-type X-ray diffraction
The molecular interaction (hydrogen bonding between
starch chains) after cooling of the gelatinized starch paste
Native RS
has been called retrogradation (Hoover 2001). Rice, maize,
RS+15% GTP
and potato starches all contained a small quantity of
RS+10% GTP
proteins that ease the retrogradation. Shimamura and Hara RS+5% GTP
(1992) reported that in TP combined with protein, the RS+0% GTP
"B" type "V" type
combining may inhibit occurring of retrogradation. It is the
other most important reason that GTPs contain highly Native MS
reactive hydroxyl (OH) groups (Wu et al. 2009), the OH MS+15% GTP
groups of GTPs interact with the OH groups of rice, maize, MS+10% GTP
and potato starches to form a hydrogen bond that interferes MS+5% GTP
MS+0% GTP
with the alignment of starch polymer chains (Beta and "B" type "V" type
Corke 2004). Phosphate monoester in potato starch inter-
acts with the OH groups of GTPs, which decreases the Native PS
effect of GTPs on preventing the retrogradation of potato PS+15% GTP
starch. PS+10% GTP
PS+5% GTP
PS+0% GTP
Recrystallization Analysis "B" type

The information on the starch granule final recrystallization 5 10 15 20 25 30 35

was investigated using XRD to further prove the effect of Diffraction angle (2θ)
GTPs on the retrogradation. The results are shown in Fig. 1.
Fig. 1 X-ray diffraction of native starch and gelatinized starch gels
Based on the XRD pattern, the crystalline region of starches containing different concentrations of GTPs after 10 days of storage at
granules added to GTPs was significantly affected. The 4 °C. RS rice starch, MS maize starch, PS potato starch
Food Bioprocess Technol (2013) 6:2177–2181 2181

pattern. However, retrograded rice and maize starches with Kim, J. O., Kim, W. S., & Shin, M. S. (1997). A comparative of rice
starch and α-amylase study on retrogradation gels by DSC, X-ray
0%, 5%, 10%, and 15% GTPs does not only gave a B-type
methods. Starch, 49, 71–75.
XRD pattern at 17° but also V structures (peak at 20°). Kuakpetoon, D., & Wang, Y. J. (2001). Characterization of different
starches oxidized by hypochlorite. Starch, 53, 211–218.
Li, W., Bai, Y., Mousaa, S. A. S., Zhang, Q., & Shen, Q. (2011).
Effect of high hydrostatic pressure on physicochemical and
Conclusions
structural properties of rice starch. Food and Bioprocess
Technology. doi:10.1007/s11947-011-0542-6.
It was shown that adding GTPs could significantly inhibit McGrance, S. J., Cornell, H. J., & Rix, C. J. (1998). A simple and
the retrogradation of rice, maize, and potato starches. rapid colorimetric method for the determination of amylose in
starch products. Starch, 50, 158–163.
Considering GTPs’ ubiquity, abundance, and low cost, it Miller, J. C., & Miller, J. N. (1993). Statistics for analytical chemistry.
will be attractive to use them as an additive to add to rice, Chichester: Ellis Horwood Ltd.
maize, and potato products to simultaneously enhance Périno-Issartier, S., Human, Z., Abert-Vian, M., & Chemat, F. (2011).
quality and nutrition. In addition, the mechanism that Solvent free microwave-assisted extraction of antioxidants from
sea buckthorn (Hippophae rhamnoides) food by-products. Food
prevents retrogradation of these starches must be under-
and Bioprocess Technology, 2011(4), 1020–1028.
stood at the molecular level, and it must be confirmed Rodríguez-Sandovala, E., Fernández-Quinterob, A., Cuvelierc, G.,
whether this is due to the highly reactive hydroxyl radical Relkinc, P., & Bello-Pérezd, L. A. (2008). Starch retrogradation
of GTPs acting with starches to form a hydrogen bond that in cassava flour from cooked parenchyma. Starch, 60, 174–180.
Shimamura, T. & Hara, Y. (1992). Method of inhibiting and treating
prevents the reassociation of starch chains.
infection caused by influenza virus. US Patent 5, 137
Wang, R., & Zhou, W. (2004). Stability of tea catechins in the bread
Acknowledgments This work was supported by the National making process. Journal of Agricultural and Food Chemistry, 52,
Natural Science Foundation (Grant No. 31050012) and the Special 8224–8229.
Fund for Agro-scientific Research in the Public Interest of China Wu, Y., Chen, Z. X., Li, X. X., & Li, M. (2009). Effect of tea
(Grant No. 200903043–2). polyphenols on the retrogradation of rice starch. Food Research
International, 42, 221–225.
Wu, Y., Chen, Z. X., Li, X. X., & Wang, Z. J. (2010). Retrogradation
properties of high amylose rice flour and rice starch by physical
modification. LWT—Food Science and Technology, 43, 492–497.
References
Xiao, H., Lin, Q., Liu, G. Q., Liu, Z., Li, L., & Yu, F. (2011). Production
of maltose syrup by enzymatic conversion of rice starch. Food and
Beta, T., & Corke, H. (2004). Effect of ferulic acid and catechin on Bioprocess Technology. doi:10.1007/s11947-011-0681-9.
sorghum and maize starch pasting properties. Cereal Chemistry, Yu, S., Ma, Y., Menager, L., & Sun, D. W. (2010). Physicochemical
81, 418–422. properties of starch and flour from different rice cultivars. Food
Chung, H. J., Woo, K. S., & Lim, S. T. (2004). Glass transition and and Bioprocess Technology. doi:10.1007/s11947-010-0330-8.
enthalpy relaxation of cross-linked corn starches. Carbohydrate Zduńczyk, Z., Frejnagel, S., Wróblewska, M., Juśkiewicz, J.,
Polymers, 55, 9–15. Oszmiański, J., & Estrella, I. (2002). Biological activity of
Feillet-Coudray, C., Sutra, T., Fouret, G., Ramos, J., Wrutniak-Cabello, polyphenol extracts from different plant sources. Food Research
C., Cabello, G., et al. (2009). Oxidative stress in rats fed a high-fat International, 35, 183–186.
high-sucrose diet and preventive effect of polyphenols: Involvement Zhou, Y. B., Wang, D. F., Zhang, L., Du, C. F., & Zhou, X. L. (2008).
of mitochondrial and MAD(P)H oxidase systems. Free Radical Effect of polysaccharides on gelatinization and retrogradation of
Biology & Medicine, 46, 624–632. wheat starch. Food Hydrocolloids, 22, 505–512.
Hoover, R. (2001). Composition, molecular structure, and physico- Zhu, F., Cai, Y. Z., Sun, M., & Corke, H. (2009). Effect of
chemical properties of tuber and root starches: A review. phytochemical extracts on the pasting, thermal, and gelling
Carbohydrate Polymers, 45, 253–267. properties of wheat starch. Food Chemistry, 112, 919–923.