15 June 2011, Volume 31 Issue 3

    

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  Protein Extraction Method Suitable for Tea Plant

  LI Qin, HUANG Jian-an, LIU Shuo-qian, LI Juan, LIU Zhong-hua

  Journal of Tea Science. 2011, 31(3): 173-178. https://doi.org/10.13305/j.cnki.jts.2011.03.014

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  In order to develop an efficient protein extraction method suitable for two-dimensional electrophoresis from tea plant, three methods (TCA/acetone precipitation, improved Tris-HCl extraction and phenol-methanol/ammonium acetate precipitation) for total protein extraction in plant were compared. The result showed that the improved Tris-HCl extraction was the best method suitable for tea leaf protein sample preparation in these three techniques and the extracts were separated by two-dimensional electrophoresis. Consequently, 1117±9.89 protein spots were found and their isoelectric points were found to lie between pI5~7. The relative molecular weight of proteins was between 15.00~95.00kD. The improved Tris-HCl extraction could get the highest resolution of two dimensional electrophoresis and high yield of protein, and it could meet the requirement for the research on tea proteomecis.
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  Research on Fast Measurement of Green Tea Polyphenols Content Based on the Electrochemistry Method

  ZHU Dan, XU Hai-rong, Martial Geiser, Mario Bernabei, Wilfried Andlauer

  Journal of Tea Science. 2011, 31(3): 179-186. https://doi.org/10.13305/j.cnki.jts.2011.03.004

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  The possibility of electrochemistry-cyclic voltammetry (CV) method applied on the fast measurement of green tea polyphenols content was investigated. Three kinds of working electrodes, silver electrode, carbon paste electrode and glassy carbon electrode, were designed and self-made for that purpose. The results showed that the silver electrode was not suitable for the content determination. A linear relationship between the anodic current and the polyphenols concentration was obtained using the carbon paste electrode or the glassy carbon electrode. Test results of green tea samples indicated that the repeatability and accuracy of the glassy carbon electrode were better than the carbon paste electrode. Compared with the national standard method, the electrochemistry method costs much less time. This research confirmed the feasibility of this method and gave a new idea to develop a fast, simple and low-cost way to detect polyphenols in tea.
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  Preliminary Study on Fractions and Correlative Properties of Theabrownin from Pu-erh Tea

  YANG Xin-he, WANG Li-li, HUANG Jian-an, WU Wen-liang, LIU Zhong-hua

  Journal of Tea Science. 2011, 31(3): 187-194. https://doi.org/10.13305/j.cnki.jts.2011.03.006

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  The water extract of pu-erh tea was extracted with chloroform, ethyl acetate, n-butanol in turn. The remaining water pigment-theabrownin obtained was installed in hydroxypropyl glucan gel (sephadex LH-20) column with acetone-aqueous solution for elution and the fractions of theabrownin were collected respectively and their physico-chemical and spectral properties were studied preliminarily. The results indicated that the 40% acetone-aqueous solution was the optimum eluent and flow was 0.05BV/h and classification of theabrownin was best in presenting 6 obvious strips in column, with difference of their chromatism parameter values, total reducing power, pH value, conventional component, UV and IR spectroscopy scanning, and stability etc. The study further demonstrated complexity of compositions and properties of theabrownin. At the same time, this investigation laid a foundation for the further research of bioactivities and chemical essence of theabrownin.
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  Production of Xylose from Tea Seed Shellswith Acid Hydrolysis

  ZHENG Sheng-hong, LI Da-xiang, FANG Shi-hui, TANG Zhi-jing

  Journal of Tea Science. 2011, 31(3): 195-200. https://doi.org/10.13305/j.cnki.jts.2011.03.002

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  In order to produce xylose from tea seed shells, The method of hydrolysis of tea seed shells under preeure was used, and the dilute hydrochloric acid as a catalyst, and the ultrasonic waveas the assisted method. The xylose was monitored by 3,5-dinitrosalicylic acid (DNS) colorimetric method. The hydrolytic factors including pre-extraction time, hydrochloric acid concentration, hydrolysis time were investigated by means of orthogonal test. Results have demonstrated that the importance of various factors were arranged in following order: time of hydrolysis>concentration of acid>particlesize>time of ultrasonic wave pre-extration. The optimum factors production were as follows: 45min, ultra-sonic wave pre-extration, ultrasonic frequency of 50kHz at 25~30℃, hydrochloric acid concentration at 0.6mol/L, particle size of seed shell at 40 mesh hydrolysis time for 3h. Under the above mentioned conditions, the highest productivity of xylose was obtained, the yield was up to 28.44%. This study provided an simple method on the comprehensive utilization of tea seed shells.
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  Comparsion of Varicol and Partial-discard Process with Traditional process to Purify ECG and EGCG from Green Tea Polyphenols Using Simulated Moving Bed Chromatography

  HUANG Yong-dong, JIANG He-yuan, JIANG Yong-wen, ZHANG Jian-yong, WANG Bin, BAI Yan, WEN Zhi-hao, WANG Wei-wei

  Journal of Tea Science. 2011, 31(3): 201-210. https://doi.org/10.13305/j.cnki.jts.2011.03.007

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  With C₁₈ bonded silica as stationary phase, ethanol and water (20︰80, V/V) as mobile phase, tea polyphenols were separated by traditional Simulated Moving Bed, Varicol process and Partial-discard process. Column configuration, switching time, feed flow rate, elution flow rate were optimized respectively in order to obtain epicatechin gallate (ECG) and epigallocatechin gallate(EGCG)with high purity and high recovery. Varicol process could be employed to separate 98% green tea polyphenols (TP98) to obtain ECG with 91.33% purity and 91.41% recovery. Partial-discard process could be adopted to separate TP98 to obtain EGCG with 90.12% purity and 97.83% recovery. Compared with traditional process, Varicol process has the advantages of higher purity and recovery of target product, while Partial-discard process shows the advantages of increasing the concentration of raffinate and the purity of target product.
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  RP-HPLC Determination of Seventeen Free Amino Acids in Tea with O-phthalaldehyde Precolumn Derivation

  YANG Wei, XIAN Shu, LI Da-xiang, WAN Xiao-chun

  Journal of Tea Science. 2011, 31(3): 211-217. https://doi.org/10.13305/j.cnki.jts.2011.03.008

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  In this paper, a reversed-phase high performance liquid chromatography combined with precolumn derivatization with O-phthalaldehyde and ?uorescence detection was set up for the qualitatively and quantitatively analysis of seventeen kinds of free amino acids in tea. The RSD of the retention time and peak area of the 17 kinds of free amino acids were 0.02%~0.70% and 0.11%~1.15%, respectively. With validation, this method was proved to be accurate, stable and reproducible for the analysis of free amino acids in tea.
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  Study on Extraction and SeparationProcess of EGCG3˝Me

  LUO Zheng-fei, JIN Xiao-fang, GONG Zheng-li, LIANG Min-zhi, WANG Yi, WANG Li-bo, CHEN Ji-wei

  Journal of Tea Science. 2011, 31(3): 218-224. https://doi.org/10.13305/j.cnki.jts.2011.03.003

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  ：The extraction and separation process of EGCG3˝Me was studied by HPLC, polyamide resin and HPTLC methods. According to the orthogonal experimental design, the optimum extracting conditions of EGCG3˝Me were as follows: alcohol concentration 70%, reflux time 30 min, reflux temperature 80℃ and solid-liquid ratio 1:10. And the yield of EGCG3˝Me was 0.89%. Large amount of EGCG3˝Me was found in the elution of alcohol concentration 40% and 60%. With elution of methanol: chloroform (7:2), the high purity EGCG3˝Me was obtained by further separation and purification of 100~120 mesh polyamide resin.
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  Studies on the Changes of Components of GreenTea by Mixed Liquid Fermentation

  LIU Wen-yao, CHEN Zhong-zheng, LI Bin, FANG Xiang

  Journal of Tea Science. 2011, 31(3): 225-229. https://doi.org/10.13305/j.cnki.jts.2011.03.009

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  In this study, Aspergillus niger H-15 and yeast stain P1,the dominant microorganisms, isolated from Puer tea were used to ferment green tea in liquid. Results showed that the changes of active components during liquid fermentation were different from those of solid-state fermentation. During liquid fermentation, tea polyphenols and caffeine showed a small dynamic change. Content of amino-acid decreased from 0.58mg/mL to 0.14mg/mL, soluble sugar decreased from 13.38mg/mL to 11.34mg/mL, and pH droped from 4.93 to 3.25. Ester catechins were degraded and could not be detected by HPLC after 40h, and the non-galloylated catechins, such as EGC、DL-C、EC and GA increased significantly, their increase rate up to 589.9%、26.5%、139.9% and 219.4% respectively.
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  Effect of Ca²⁺ on the Chemical Components and Sensory Quality of Extracted Green Tea Infusion

  XU Yong-quan, CHEN Gen-sheng, ZHONG Xiao-yu, WANG Fang, YANG Yu-zhou, YUAN Hai-bo, YIN Jun-feng

  Journal of Tea Science. 2011, 31(3): 230-236. https://doi.org/10.13305/j.cnki.jts.2011.03.010

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  The effect of different Ca²⁺ concentration（0~100mg/L）on the chemical components and sensory quality of extracted green tea infusion was investigated. The results showed that, with the increasing of Ca²⁺ concentration, the turbidity of tea infusion increased, pH decreased; the contents of total solids and free amino acids increased, the contents of tea polyphenols and protein decreased, but there was no significant difference in the contents of caffeine and flavones; the contents of EGC increased, the contents of C, EGCG, GCG, ECG and CG decreased, but there was no significant difference in the contents of EC; the contents of Al³⁺, Cu²⁺, Na⁺, Zn²⁺ decreased, and the contents of K⁺, Mg²⁺, Mn²⁺ increased; tea infusion turned turbid, ripe and sulks odour enhanced, and bitter and astringency strengthened. The effect of Ca²⁺ on tea infusion quality maybe arisen by participating in tea cream formation, inducing epimerisation of catechins and appearing decomposition reaction.
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  Uptake and Accumulation Characters of Lead in Tea Plants (Camellia sinensis) Grown in Hydroponics

  ZHAO Xian-min, WANG Yan-xia, DU Xiao, LI Pin-wu

  Journal of Tea Science. 2011, 31(3): 237-246. https://doi.org/10.13305/j.cnki.jts.2011.03.016

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  Lead (Pb) concentration was investigated in tea plants (Camellia sinensis) grown in hydroponics by the FAAS method to understand the Pb uptake, accumulation and distribution characters in tea plants. The results showed that Pb ion can be uptaken and accumulated by roots in a large quantity, transported through stems along with concentration gradient, and redistributed. Cultured in 2500mg/L Pb solution or sprayed the same solution on leaves (cultured in distilled water), Pb concentration in roots were 8.00×10⁴mg/kg and 8.38×10³mg/kg, respectively, in leaves were 551.79mg/kg and 1.03×10⁴mg/kg respectively. Even such high accumulation the tea plants were still alived. At the range of 800~2000mg/L Pb solution, tea plants were gradually damaged; in the early time, absorbing roots became transparent, and then turned brown, and leaves were defoliated in the latter period. Tea plant cultured under the 300~800mg/L Pb solution for 35d, these plants grew well, and no damaged symptoms can be observed. Results indicated that tea plants possessed a strong ability of Pb uptake, accumulation and tolerance.
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  Effects of Fu Brick Tea and Its Formula on the Content of TG in L-02 Steatotic Hepatocytes

  XU Xiao-jiang, HUANG Jian-an, XIAO Li-zheng, YUAN Yong, XIAO Wen-jun

  Journal of Tea Science. 2011, 31(3): 247-254. https://doi.org/10.13305/j.cnki.jts.2011.03.011

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  Using 1mL/L of 20% medical fat emulsion to process L-02 liver cells for 48h in order to replicate non-alcoholic fatty degeneration of liver cells, and then adding different concentrations of extracts and its formulas to process separately for 24h and 48h, automatic biochemical analyzer detects the level of triglycerides (TG) in liver cells, investigate the effects of Fu Brick Tea extraction and the two formulas on the content of TG in L-02 Steatotic Hepatocytes. Results showed that Fu brick tea extraction and formula 1 can significantly lower TG levels in fatty liver cells(P<0.05), while comparing with the Xue Zhi Kang group, those two groups presented no significant difference (P>0.05). Accordingly, Fu brick tea and formula 1 reflect a wide prospect of development and application.
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  The Antagonistic Effects of Green Tea Polyphenols on the Malformations of the Fertilized Eggs of Japanese Medaka (Oryzias latipes) Induced by Benzo[a]pyrene

  SONG Chuan-kui, XIAO Bin, WANG Yan-li, YU You-ben, ZHOU Tian-shan

  Journal of Tea Science. 2011, 31(3): 255-263. https://doi.org/10.13305/j.cnki.jts.2011.03.001

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  The effects of green tea polyphenols on the malformations of the fertilized eggs of Japanese medaka (Oryzias latipes) induced by benz[a]pyrene was investigated by using the biological model of spawning Japanese medaka (Oryzias latipes) with the method of intraperitoneal injection, and the real-time PCR was used to detection the mRNA expression of CYP enzymes (CYP1A1, CYP1B, CYP2A, CYP2C, CYP2D, CYP3A) and the GST gene (mGST, GST-a) in the liver medaka. The results showed that the malformation rate of the fertilized eggs of Japanese medaka in the treatment of injection of benzo[a]pyrene which was significantly higher than the DMSO control. The malformation rate in the treatments of tea polyphenols was decreased, the treatments except the lowest dose of tea polyphenols (20μg/g BaP +2μg/g TP) were significantly different compared to the BaP treatment. Real-time quantitative PCR results showed that the benzo[a]pyrene model group slightly induces the CYP1A1, CYP1BCYP2A, CYP2C, CYP2D mRNA expression and significantly inhibits the mGST and GST-a mRNA expression of the female medaka, while tea polyphenols can significantly inhibit the expression CYP1A1 mRNA gene and glutathione-S transferase (GST) gene mGST and GST-a. The antagonistic mechanism of green tea polyphenols on the malformation of the fertilized eggs of Japanese medaka induced by benzo[a]pyrene might be correlated with the inhibition of CYP1A1 mRNA gene expression and the acceleration of the excretion of metabolic product of benzopyrene.
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  Organization of Chinese Tea Industry:Performance and Policy

  SU Zhu-cheng

  Journal of Tea Science. 2011, 31(3): 264-272. https://doi.org/10.13305/j.cnki.jts.2011.03.012

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  The organization of Chinese tea industry, its performance and policy, were discussed in this paper. The present organizational structure of Chinese tea industry showed some features such as small householder of tea garden, no-monopoly competition, market division between primary and refining production, processing for exportation according to giving samples and so on. From perspective of other major tea-production countries, the organizational structure was characterized by vertical proportional share-distribution in Sri Lanka, cooperation institution of farmers in Japan and exportation of black tea by auction market, which had some significant implication for us. It was suggested that,targeting the market entry of small householder of tea garden and integration of primary and refining production, industry polices should focused on innovation of institution and technology. Besides, transformation of mode of exportation transaction should also be explored.
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  Analysis on Development Scale ofChineseTea Industry in 2020

  JIANG Yong-wen, CHEN Xiao-xiong, ZHU Jian-miao, YANG Shuang-xu

  Journal of Tea Science. 2011, 31(3): 273-282. https://doi.org/10.13305/j.cnki.jts.2011.03.013

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  This paper analyzed achievements and developing factors of Chinese tea industry since the 21^th century and made predictive parsing to the external environment, supplying ability and demand of tea in five to ten years. It was considered, though the development of Chinese tea industry was restricted by market demand and production factors, that it would have a relative broad space for development. Chinese tea industry could develop steadily in future on the conditions to solve its present issues, grasp market chance, improve competitive ability and quicken the pace of upgrade. It was predicted that the total tea output of China would be 1785000 tons in 2015 and 2200000 tons in 2020.