普洱茶多酚氧化产物的提取分离、成分分析及活性评价
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摘要
普洱茶属于后发酵茶，是我国特有的一类茶，有很大的市场发展潜力。本实验首次以云南野生大茶树原料制作的普洱茶为材料，对普洱茶多酚氧化产物的提取分离工艺进行了研究，并探索其降血脂、抗氧化的功能，旨在系统、全面地研究和科学地评价普洱茶，为普洱茶深加工技术改进和产业化发展提供理论支持，为普洱茶功能食品的开发提供科学依据。
1普洱茶多酚氧化产物浸提工艺的研究及其提取物主要品质成分分析
以普洱茶为原料，研究溶剂pH值、浸提温度、时间、料液比对茶褐素(TBS)提取率的影响。结果表明，溶剂pH值、浸提温度、时间、料液比等都显著影响TBS的浸提效果，其中料液比、浸提温度、浸提时间达到了极显著水平，其影响效果的重要性依次为：料液比＞浸提温度＞浸提时间＞溶剂pH值。同时得到茶褐素浸提的优化条件为：温度100℃，溶剂pH=7，料液比1：30，时间20min。
发酵是影响普洱茶品质成分变化的主要原因。普洱茶所含的化学成分与其原料相比差异较大，普洱茶中茶多酚、没食子酸、氨基酸(尤其是茶氨酸)含量远低于晒青绿茶，茶黄素、茶红素、茶褐素含量以及可溶性糖含量则高于晒青绿茶。茶多酚在普洱茶中主要以简单儿茶素的形式存在，而在晒青绿茶中则主要以酯型儿茶素形式存在。主要品质成分没食子酸、茶氨酸和咖啡碱在普洱茶多酚氧化产物中的分布差异较大。没食子酸主要存在于TFS，茶氨酸主要存在于TRS和TBS，咖啡碱主要存在于TFS和TRS。
2普洱茶及其多酚氧化产物的降血脂功能研究
以晒青绿茶提取物(GE)为对照，分别以高、中、低不同剂量的普洱茶水提物(PE)、茶黄素(TFS)、茶红素(TRS)和茶褐素(TBS)灌胃高脂模型小鼠，40天后眼眶取血、分离肝组织，测血脂指标。结果表明：
(1)普洱茶及其多酚氧化产物能有效调节预防性高脂小鼠血清血脂水平。TBS中高剂量、PE各剂量对高脂小鼠血清TC含量有极显著的降低作用。TRS、TBS、PE各剂量组都能极显著的降低小鼠血清TG含量，普洱茶效果好于其多酚氧化产物。除PE中低剂量组外其他供试品各剂量组均能极显著的增加HDL-C含量，TFS、TRS组效果最好。
(2)普洱茶及其多酚氧化产物对预防型高脂小鼠肝匀浆血脂调节作用主要体现于TG、TC含量的降低。TRS各剂量组，PE各剂量组能有效预防肝匀浆中TC升高，除TRS高剂量外其余各组都显著降低肝匀浆中TG含量，且茶汤提取物效果较其他单独多酚氧化产物效果显著。小鼠肝脏切片显示，灌胃组能有效防止高脂肝细胞正常组织结构消失程度。
(3)普洱茶原料晒青绿茶降低血脂的效果优于普洱茶。表现在晒青绿茶组TC、TG含量最低，并且比普洱茶及其多酚氧化产物明显降低血清TC含量。
3普洱茶及其多酚氧化产物的抗氧化活性研究
按上述同样的方法分析普洱茶及其多酚氧化产物的体内外抗氧化作用。结果表明：
(1)普洱茶及其多酚氧化产物能显著增强预防型高脂小鼠血清及肝脏的抗氧化功能。PE组，TBS高剂量组、TFS中剂量组能极显著提高血清SOD活力，各供试组均能极显著地提高肝脏SOD活力。各供试组都能极显著的降低血清MDA含量，其中以TRS对小鼠血清MDA生成影响最显著，但在肝脏中仅PE高剂量组能有效降低MDA含量。
(2)普洱茶及其多酚氧化产物在体外对O_2·~-自由基有较强的清除活性。其清除O_2·~-自由基的成份主要集中在TFS部分，清除能力大小依次为TFS＞TRS＞TBS＞PE。
(3)绿茶的抗氧化活性优于普洱茶。主要表现在体内能显著提高预防性高脂小鼠SOD活力，降低MDA含量，体外对O_2·~-自由基有较强的清除能力。
Pu'er tea is post fermented tea, a special tea with much market-developingpotential in China. The method of extraction and isolation of polyphenol Oxides fromPu'er tea was studied in my project; also we studied its effect on rat's blood lipids andantioxidant capacity. This is the first experiment on the material of Yunnan wild tearaw. In order to study it comprehensively, evaluate it scientifically, and provideintensive processing technology and industrial development theories. Also we mayguide the development of functional foods, the consumers' choice and the teaconsumer market.
1. Study on extraction technology of polyphenol oxides and main qualitycomponent from Pu'er tea
The influence of pH value of solvent, temperature, heating time and ratio of teato solvent on extraction rate of TBS was studied. The results showed that allparameters affected the extraction rate noticeably, and the effects of last threeparameters was extremely remarkable. The order of importance was: ratio of tea tosolvent＞temperature＞heating time＞pH value of solvent. The optimized parameterswere: using pH=7 water solution as extract solvent, the ratio of tea to solvent was 1:30,and extracting at 100℃for 20min.
Fermentation was the main influence factor of Pu'er tea quality. Pu'er teachemical contents had much difference with its material raw. Tea polyphenol, gallicacid, amino acids (in particular theanine) was much lower in Pu'er tea, while TFS,TRS, TBS and soluble sugar content was higher than the drying green tea. The maincatechins of Pu'er tea were simple catechins, while the drying green tea catechinswere complex catechins. The distribution of gallic acid, theanine and caffeine in Pu'ertea polyphenol oxides was quit difference. Gallic acid was found primarily in the TFS,while theanine in the TRS and TBS, and Caffeine in the TFS and TRS.
2. Study on Pu'er tea and its polyphenol oxides on reducing blood lipids
Using the drying green tea extraction (GE) as a control, hyperlipidemia modelmice were intragastric Infusion with different dose of TFS, TRS, TBS, PE and GE for40 days. After last administration, he mice were fasted at least 10 hours, and then theywere sacrificed by cervical dislocation, serum was prepared from blood obtained ineye vein cluster and TC,TG,HDL-C,MDA and SOD were assayed; The liver washomogenized in physiology saline and TC, TG, MDA and SOD were assayed. Theresults were showed:
(1) Pu'er tea and its polyphenol oxides can be effectively regulated preventivehigh fat mice serum lipid level. TBS with medi-dose, high-dose and each dose of PE could remarkably decrease the content of TC; TRS, TBS and PE at all doses couldsignificantly lower the content of TG, the effect of Pu'er tea was better than itspolyphenol oxides; All the tasted samples could increase HDL-C contents of serumsignificantly except PE medi-dose and low dose, and the effect of TFS, TRS wasbetter than others.
(2)The regulative effect on high-fat mice liver of Pu'er tea and its polyphenoloxides was mainly reflected in the decreased of TG, TC content; Each dose of TRSand PE could prevent the increase of TC content effectually on the liver of high-fatmice, and all doses of tasted samples except high dose of TRS could significantlyreduced liver triglyceride content of high-fat diet mice, and the effect of PE was betterthan its polyphenol oxides. The liver biopsy of high-fat group showed that: theexperimental groups can be effective in preventing the disappear extent of high-cellsof normal liver structure.
(3)The Green tea had more effect on reducing blood lipids than Pu'er tea. As theGE group had the lowest content of TC and TG, and GE can reduce the TC contentmuch more significantly than other samples.
3. The effect of Pu'er tea and its polypheuol oxides on antioxidant ability
The antioxidant activity of TFS, TRS, TBS, TA and green tea extracts wasstudied. The results showed:
(1)Pu'er tea and its polyphenol oxides can significantly enhance the preventivehigh-fat mouse serum and liver antioxidant function. PE group, TBS medi-dose,high-dose groups, TFS medi-dose group could significantly increase the SOD activityof serum, and all the samples could significantly raise the SOD activity of liver. Allthe test groups can significantly reduce the content of MDA in serum, and TRS hadthe most significant impact, but only high-dose of PE could effectively reduce thecontent of MDA significantly in the liver.
(2)Pu'er tea and its polyphenol had strong activity in vitro oxides of O_2·-radicalscavenging. The removal of oxygen-free radicals ingredients mainly concentrated inthe TFS, the order of removing ability was: TFS＞TRS＞TBS＞PE.
(3) The antioxidant activity of Green tea was better than the Pu'er tea. Mainlybecause of that it can significantly improve the SOD activity and lower MDA contentof preventive high fat mice in the body, and its strong scavenging capacity of O_2·~-.
1普洱茶多酚氧化产物浸提工艺的研究及其提取物主要品质成分分析
以普洱茶为原料，研究溶剂pH值、浸提温度、时间、料液比对茶褐素(TBS)提取率的影响。结果表明，溶剂pH值、浸提温度、时间、料液比等都显著影响TBS的浸提效果，其中料液比、浸提温度、浸提时间达到了极显著水平，其影响效果的重要性依次为：料液比＞浸提温度＞浸提时间＞溶剂pH值。同时得到茶褐素浸提的优化条件为：温度100℃，溶剂pH=7，料液比1：30，时间20min。
发酵是影响普洱茶品质成分变化的主要原因。普洱茶所含的化学成分与其原料相比差异较大，普洱茶中茶多酚、没食子酸、氨基酸(尤其是茶氨酸)含量远低于晒青绿茶，茶黄素、茶红素、茶褐素含量以及可溶性糖含量则高于晒青绿茶。茶多酚在普洱茶中主要以简单儿茶素的形式存在，而在晒青绿茶中则主要以酯型儿茶素形式存在。主要品质成分没食子酸、茶氨酸和咖啡碱在普洱茶多酚氧化产物中的分布差异较大。没食子酸主要存在于TFS，茶氨酸主要存在于TRS和TBS，咖啡碱主要存在于TFS和TRS。
2普洱茶及其多酚氧化产物的降血脂功能研究
以晒青绿茶提取物(GE)为对照，分别以高、中、低不同剂量的普洱茶水提物(PE)、茶黄素(TFS)、茶红素(TRS)和茶褐素(TBS)灌胃高脂模型小鼠，40天后眼眶取血、分离肝组织，测血脂指标。结果表明：
(1)普洱茶及其多酚氧化产物能有效调节预防性高脂小鼠血清血脂水平。TBS中高剂量、PE各剂量对高脂小鼠血清TC含量有极显著的降低作用。TRS、TBS、PE各剂量组都能极显著的降低小鼠血清TG含量，普洱茶效果好于其多酚氧化产物。除PE中低剂量组外其他供试品各剂量组均能极显著的增加HDL-C含量，TFS、TRS组效果最好。
(2)普洱茶及其多酚氧化产物对预防型高脂小鼠肝匀浆血脂调节作用主要体现于TG、TC含量的降低。TRS各剂量组，PE各剂量组能有效预防肝匀浆中TC升高，除TRS高剂量外其余各组都显著降低肝匀浆中TG含量，且茶汤提取物效果较其他单独多酚氧化产物效果显著。小鼠肝脏切片显示，灌胃组能有效防止高脂肝细胞正常组织结构消失程度。
(3)普洱茶原料晒青绿茶降低血脂的效果优于普洱茶。表现在晒青绿茶组TC、TG含量最低，并且比普洱茶及其多酚氧化产物明显降低血清TC含量。
3普洱茶及其多酚氧化产物的抗氧化活性研究
按上述同样的方法分析普洱茶及其多酚氧化产物的体内外抗氧化作用。结果表明：
(1)普洱茶及其多酚氧化产物能显著增强预防型高脂小鼠血清及肝脏的抗氧化功能。PE组，TBS高剂量组、TFS中剂量组能极显著提高血清SOD活力，各供试组均能极显著地提高肝脏SOD活力。各供试组都能极显著的降低血清MDA含量，其中以TRS对小鼠血清MDA生成影响最显著，但在肝脏中仅PE高剂量组能有效降低MDA含量。
(2)普洱茶及其多酚氧化产物在体外对O_2·~-自由基有较强的清除活性。其清除O_2·~-自由基的成份主要集中在TFS部分，清除能力大小依次为TFS＞TRS＞TBS＞PE。
(3)绿茶的抗氧化活性优于普洱茶。主要表现在体内能显著提高预防性高脂小鼠SOD活力，降低MDA含量，体外对O_2·~-自由基有较强的清除能力。
Pu'er tea is post fermented tea, a special tea with much market-developingpotential in China. The method of extraction and isolation of polyphenol Oxides fromPu'er tea was studied in my project; also we studied its effect on rat's blood lipids andantioxidant capacity. This is the first experiment on the material of Yunnan wild tearaw. In order to study it comprehensively, evaluate it scientifically, and provideintensive processing technology and industrial development theories. Also we mayguide the development of functional foods, the consumers' choice and the teaconsumer market.
1. Study on extraction technology of polyphenol oxides and main qualitycomponent from Pu'er tea
The influence of pH value of solvent, temperature, heating time and ratio of teato solvent on extraction rate of TBS was studied. The results showed that allparameters affected the extraction rate noticeably, and the effects of last threeparameters was extremely remarkable. The order of importance was: ratio of tea tosolvent＞temperature＞heating time＞pH value of solvent. The optimized parameterswere: using pH=7 water solution as extract solvent, the ratio of tea to solvent was 1:30,and extracting at 100℃for 20min.
Fermentation was the main influence factor of Pu'er tea quality. Pu'er teachemical contents had much difference with its material raw. Tea polyphenol, gallicacid, amino acids (in particular theanine) was much lower in Pu'er tea, while TFS,TRS, TBS and soluble sugar content was higher than the drying green tea. The maincatechins of Pu'er tea were simple catechins, while the drying green tea catechinswere complex catechins. The distribution of gallic acid, theanine and caffeine in Pu'ertea polyphenol oxides was quit difference. Gallic acid was found primarily in the TFS,while theanine in the TRS and TBS, and Caffeine in the TFS and TRS.
2. Study on Pu'er tea and its polyphenol oxides on reducing blood lipids
Using the drying green tea extraction (GE) as a control, hyperlipidemia modelmice were intragastric Infusion with different dose of TFS, TRS, TBS, PE and GE for40 days. After last administration, he mice were fasted at least 10 hours, and then theywere sacrificed by cervical dislocation, serum was prepared from blood obtained ineye vein cluster and TC,TG,HDL-C,MDA and SOD were assayed; The liver washomogenized in physiology saline and TC, TG, MDA and SOD were assayed. Theresults were showed:
(1) Pu'er tea and its polyphenol oxides can be effectively regulated preventivehigh fat mice serum lipid level. TBS with medi-dose, high-dose and each dose of PE could remarkably decrease the content of TC; TRS, TBS and PE at all doses couldsignificantly lower the content of TG, the effect of Pu'er tea was better than itspolyphenol oxides; All the tasted samples could increase HDL-C contents of serumsignificantly except PE medi-dose and low dose, and the effect of TFS, TRS wasbetter than others.
(2)The regulative effect on high-fat mice liver of Pu'er tea and its polyphenoloxides was mainly reflected in the decreased of TG, TC content; Each dose of TRSand PE could prevent the increase of TC content effectually on the liver of high-fatmice, and all doses of tasted samples except high dose of TRS could significantlyreduced liver triglyceride content of high-fat diet mice, and the effect of PE was betterthan its polyphenol oxides. The liver biopsy of high-fat group showed that: theexperimental groups can be effective in preventing the disappear extent of high-cellsof normal liver structure.
(3)The Green tea had more effect on reducing blood lipids than Pu'er tea. As theGE group had the lowest content of TC and TG, and GE can reduce the TC contentmuch more significantly than other samples.
3. The effect of Pu'er tea and its polypheuol oxides on antioxidant ability
The antioxidant activity of TFS, TRS, TBS, TA and green tea extracts wasstudied. The results showed:
(1)Pu'er tea and its polyphenol oxides can significantly enhance the preventivehigh-fat mouse serum and liver antioxidant function. PE group, TBS medi-dose,high-dose groups, TFS medi-dose group could significantly increase the SOD activityof serum, and all the samples could significantly raise the SOD activity of liver. Allthe test groups can significantly reduce the content of MDA in serum, and TRS hadthe most significant impact, but only high-dose of PE could effectively reduce thecontent of MDA significantly in the liver.
(2)Pu'er tea and its polyphenol had strong activity in vitro oxides of O_2·-radicalscavenging. The removal of oxygen-free radicals ingredients mainly concentrated inthe TFS, the order of removing ability was: TFS＞TRS＞TBS＞PE.
(3) The antioxidant activity of Green tea was better than the Pu'er tea. Mainlybecause of that it can significantly improve the SOD activity and lower MDA contentof preventive high fat mice in the body, and its strong scavenging capacity of O_2·~-.
引文
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25．杨贤强，叶立扬，贾之慎．茶叶天然抗氧化剂的提取．浙江农业大学学报，1992，18(10)：19-22
26．周红杰．普洱茶的品质特点及健身作用．农牧产品开发，1997(2)：38
27．周红杰，秘鸣，韩俊，李家华，艾田．普洱茶的功效及品质形成机理研究进展．茶叶，2003，29(2)：75-77
28．朱建中，邵建利．统计应用软件EXCEL和SAS．上海：上海财经大学出版社，2002
29．钟萝．茶叶品质理化分析．上海：上海科技出版社，1989
30．良边文久．A Simple Preparation Chromatographic Separation of Green tea Catechins. Nippon Kaishi Nigeikagaku, 1989, 63 (4): 845-847.
31．龚加顺，周红杰．云南晒青绿毛茶的微生物固态发酵及成分变化研究．茶叶科学，2005，25(4)：300～306
32．吴文华．晒青毛茶普洱茶降血脂功能比较．中国茶叶·试验研究2005．1
33．周斌星，高林瑞．普洱茶茶色素的研究进展．茶叶科学技术，2005(3)
34．张冬英，施兆鹏，刘亚林．普洱茶药理作用研究进展．福建茶业．饮茶与健康，2005(4)43-44
35．李春美，谢笔钧．儿茶素氧化聚合产物药理作用研究概况．茶叶2001，27(1)：28-3
36．王坤波，刘仲华，黄建安．茶黄素的提取分离与纯化研究进展．湖南农业大学学报(自然科学版)2002，18(4)：355-35
37．杜琪珍，江和源．茶色素的药理及功效．中国茶叶，1997，19(5)：42-43
38．楼福庆．茶色素与动脉粥样硬化症．中国茶叶，1987，(2)：28-30
39．江和源，程启坤，杜琪珍．红茶中的茶黄素．中国茶叶，，1998，20(3)：18-20
40．程启坤．红茶中的茶黄素．国外农学—茶叶，1983(1)：1-14
41．江和源，程启坤，杜琪珍．高速逆流色谱在茶黄素分离上的应用．茶叶科学，2000，20(1)：40-44
42．竹尾忠一．茶黄素的分离与定量．茶叶技术研究，1973，45：46
43．楼福庆，杨祖才，袁伟龙．茶色素对家兔实验性动脉粥样硬化和人纤维蛋白原增多症的作用．中华医学杂志，1983，63(10)：632
44．曹进，赵燕，刘箭卫．红茶及茶色素药理作用研究的动态．中国药学杂志，1999，634(6)：361-363
45．陈红伟．传统普洱茶与现代普洱茶．茶与饮品，茶苑，2004(1)：21-22
46．吴桂兰．复方茶色素的药理研究．中成药，46-47
47．周树红．成品普洱茶陈化机理及提质技术研究．浙江大学，2005
48．杨家林，窦薇．普洱红茶和普洱绿茶降低老年人血脂的对比观察．中国老年学杂志1997，6(17)：174
49．谢一辉，罗永明．茶色素胶囊中总多酚的紫外分光光度法测定．中国医药工业杂志，2001，32(5)：227-230
50．夏勇，徐彩菊．茶色素对实验性高脂血症大鼠血脂水平的影响浙江预防医学．2003，15 (7)：76-77
51．邵翔，杨浩，俞国华．茶提取物在心血管疾病中的应用．现代中西医结合杂志．2005，14(14)：1917-1919
52．李大祥，宛晓春．茶色素中茶黄素类的HPLC定量，茶叶科学2004，24(2)：124—128
53．郑青山，丁学庭，宋寅．茶多酚对大鼠实验性高血脂的影响．安徽医药．1997 71(2)
54．王汉生．茶多酚降血脂及抗氧化作用动物实验．华南农业大学
55．龚志华，仇灿红，肖文军．茶多酚降血脂研究进展．福建茶叶2002(4)：39-40
56．宋小鸽．茶多酚提取工艺及降脂作用的概述．黑龙江中医药，1998(6)：51-53
57．林智，吕海鹏，崔文锐．普洱茶的抗氧化酚类化学成分的研究．茶叶科学2006，26(2)：112～116
58．杨慧琴，赵斌，王江民．茶色素治疗高脂血症附150例疗效观察．邯郸医学高等专科学校学报，2000，13(6)：443-444
59．王路宏，钟永香．茶色素调节血脂代谢紊乱90例．中国新药与临床杂志，1999，11，18(6)：409—410
60. Helaine M. Alessio, Ann E. Hagerman. Consumption of green tea protects rats from exerciseinduced oxidative stress in kidney and liver. Nutrition Research 22 (2002): 1177-1188
61. J.J. Choo. Green tea reduces body fat accretion caused by high-fat diet in rats through β-adrenoceptor activation of thermogenesis in brown adipose tissue. Journal of Nutritional Biochemistry 11 (2003): 671-676
62. Ming-Hua Yang, Cheng-Hsin Wang. Green, oolong and black tea extracts modulate lipid mePEbolism in hyrlipidemia rats fed high-sucrose diet. Journal of Nutritional Biochemistry 12 (2001) 14-20
63. M. LODOVICI, C, CASALINI. Inhibition of 1, 2-dimethylhydrazine-induced oxidative DNA damage in rat colon mucosa by black tea complex polyphenols Food and Chemical Toxicology 38 (2000): 1085-1088
64. Rajesh Krishnan, Girish B. Maru Isolation and analyses of polymeric polypbenol fractions from black tea Food Chemistry 94 (2006) 331-340
65. Vasyl M. Sava, Swen-Ming Yang. Isolation and characterization of melanic pigments derived from tea and tea polypbenols.Food Chemistry 73(2001) 177-184
66. S. De Vos, R. De Schrijver. Lipid mePEbolism, intestinal fermenPEtion and mineral absorption in rats consuming black tea. Nutrition Research 23 (2003) 527-537
67. E. Skrzydlewska, J. Ostrowska, R. Farbiszewski. Protective effect of green tea against lipid roxidation in the rat liver, blood serum and the brain. Phytomedicine 9: 232-238, 2002
68. Nuria Martin-Carron, BSc, Isabel Goni, Ph.D. Reduction in serum toPEl and LDL cholesterol concentrations by a diePEry fiber and polyphenol-rich gra product in hyrcholesterolemic rats. Nutrition Research, Vol, 19, No.9, pp. 1371-1381, 1999
69. Scavenging Effects of Green Tea Polypbenols on the Oxygen Free Radicals Generated From Isolated Iscbemic Rerfused Rat Myocardium
70. Inoue M, Suzuki R, Koide T. Antioxidant, gallic acid. induces apoptosis in HL-60RG cells. Biocbem Biophys Res Commun, 1994, 204 (2): 898-904
71. Inouc M, Suzuki R, Sakaguchi N. Selective induction of cell detch in cancer cells by gallic acid. Biol Pharm Bull, 1995, 18 (11): 1526-1530
72. Inoue M, Sakaguchi N, lsuzugawa K. Role of reactive oxygen scies in gallic acid. induced apoptosis. Biol Pharm Bull, 2000, 23 (10): 1153-1157
73. Mukberjee G, Banerjee R. Biosynthesis of PEnnase and gallic acid from PEnnin rich substrates by Rhizopus oryzae and Asrgillus foetidus. Basic Microbio, 2004, 44(1): 42-8
74. Olmo T, lnoue M, Ogihara Y. Cytotoxic activity of gallic acid against liver mePEsPEsis of mastocytoma cells P815. Anticancer Res, 2001, 21 (6A): 3875-3880
75. YEN Gow-chin, CHEN Hui-yin. Antioxidant activity of tea extracts in relation to their antimuPEgenicity. J Agric Food chem., 1997, 45:30-34
76. Zhou ZH (周志宏), Yang CR (杨崇仁). Chemical constituents of crude green tea: the material of Pu'er tea in Yunnan. AcPE Bot Yunnan (云南植物研究), 2000, 22(3): 343-350
2．黄华涛．云南沱茶——高脂血症克星．茶叶，1990，1：47
3．陈海霞，谢笔钧．茶叶中有效成分的提取制备研究进展．食品研究与开发，1999，20(5)：14-16
4．邓时海．论普洱茶越陈越香．中国普洱茶文化研究．云南科技出版社，1994年第1版：147-156
5．葛蕾．苹果渣多酚浸提物及其抗氧化研．[硕士学位论文]．西北农林科技大学图书馆，2005
6．何国藩，林月蝉．普洱茶对肠段的舒缩推进运动和胃蛋白酶分泌的影响．中国茶叶，1988(2)：6-8
7．何国藩，林月蝉，徐福祥．普洱茶对人血压心率及脑血流图的影响．中国茶叶，1995(3)：27-28
8．何金柱，杨普，袁自春．温度、时间、pH值对红茶有效成分浸提率的影响．安徽农业科学．2005，33(3)：467-468
9．何晓燕．速溶普洱茶的研制．茶业机械杂志，1996(3)：13-14
10．黄桂枢．中国普洱茶文化研究．昆明：云南人民出版社，1994
11．揭国良，何普明，丁仁凤．普洱茶抗氧化特性的初步研究．茶叶，2005，31(3)：162-165
12．蒋建平．天然抗氧化剂茶多酚的提取和其在食品保鲜中的应用．[硕士学位论文]．长沙：湖南大学图书馆，2003
13．李春美，谢笔均．茶多酚及其氧化产物清除不同体系产生活性氧自由基的分光光度法研究．精细化工，2000，17(4)：241-244
14．吕海鹏．普洱茶的化学成分分析及其抗氧化活性研究．[硕士学位论文]．中国农业科学院，2005
15．赖建辉．低档茶叶及茶渣的利用．中国茶叶加工，1995，(1)：21-24．
16．罗龙新，吴小崇，傅尚文．不同产地普洱茶品质风格的比较．中国茶叶，1995(5)：8-10
17．梁名志．漫谈古今神奇普洱茶药效．云南茶叶，1996，1(2)：7-10
18．苏芳华．弘扬普洱茶文化让更多的人喜欢普洱茶．2002年中国普洱茶国际学术研讨会论文集．云南人民出版社，2002年6月第1版：48-58
19．折改梅，张香兰，陈可可，张颖君，杨崇仁．茶氨酸和没食子酸在普洱茶中的含量变化．云南植物研究，2005，27(5)：572-576
20．孙璐西，刘张惠泉．普洱茶之抗动脉硬化作用，2002年中国普洱茶国际学术研讨会论文集．云南人民出版社，2002年6月第1版：309-317
21．王树文，苏芳华．茶的故乡．云南．中国土蓄产进出口公司，云南茶叶分公司(内部资料)[C]1990
22．吴文华．普洱茶调节血脂功能评价及其生化机理的研究．[硕士学位论文]．重庆：西南农业大学图书馆，2005
23．谢笔钧，石煌，胡慰望．绿茶、乌龙茶、红茶中主要组分和酚类化合物抑制脂肪氧合酶活性和抗油脂自动氧化特性的研究．天然产物研究与开发，1994 6(4)：20—23
24．谢笑天．茶灰中提取抗氧化剂工艺研究．云南师范大学学报，1997，17(3)：50-52
25．杨贤强，叶立扬，贾之慎．茶叶天然抗氧化剂的提取．浙江农业大学学报，1992，18(10)：19-22
26．周红杰．普洱茶的品质特点及健身作用．农牧产品开发，1997(2)：38
27．周红杰，秘鸣，韩俊，李家华，艾田．普洱茶的功效及品质形成机理研究进展．茶叶，2003，29(2)：75-77
28．朱建中，邵建利．统计应用软件EXCEL和SAS．上海：上海财经大学出版社，2002
29．钟萝．茶叶品质理化分析．上海：上海科技出版社，1989
30．良边文久．A Simple Preparation Chromatographic Separation of Green tea Catechins. Nippon Kaishi Nigeikagaku, 1989, 63 (4): 845-847.
31．龚加顺，周红杰．云南晒青绿毛茶的微生物固态发酵及成分变化研究．茶叶科学，2005，25(4)：300～306
32．吴文华．晒青毛茶普洱茶降血脂功能比较．中国茶叶·试验研究2005．1
33．周斌星，高林瑞．普洱茶茶色素的研究进展．茶叶科学技术，2005(3)
34．张冬英，施兆鹏，刘亚林．普洱茶药理作用研究进展．福建茶业．饮茶与健康，2005(4)43-44
35．李春美，谢笔钧．儿茶素氧化聚合产物药理作用研究概况．茶叶2001，27(1)：28-3
36．王坤波，刘仲华，黄建安．茶黄素的提取分离与纯化研究进展．湖南农业大学学报(自然科学版)2002，18(4)：355-35
37．杜琪珍，江和源．茶色素的药理及功效．中国茶叶，1997，19(5)：42-43
38．楼福庆．茶色素与动脉粥样硬化症．中国茶叶，1987，(2)：28-30
39．江和源，程启坤，杜琪珍．红茶中的茶黄素．中国茶叶，，1998，20(3)：18-20
40．程启坤．红茶中的茶黄素．国外农学—茶叶，1983(1)：1-14
41．江和源，程启坤，杜琪珍．高速逆流色谱在茶黄素分离上的应用．茶叶科学，2000，20(1)：40-44
42．竹尾忠一．茶黄素的分离与定量．茶叶技术研究，1973，45：46
43．楼福庆，杨祖才，袁伟龙．茶色素对家兔实验性动脉粥样硬化和人纤维蛋白原增多症的作用．中华医学杂志，1983，63(10)：632
44．曹进，赵燕，刘箭卫．红茶及茶色素药理作用研究的动态．中国药学杂志，1999，634(6)：361-363
45．陈红伟．传统普洱茶与现代普洱茶．茶与饮品，茶苑，2004(1)：21-22
46．吴桂兰．复方茶色素的药理研究．中成药，46-47
47．周树红．成品普洱茶陈化机理及提质技术研究．浙江大学，2005
48．杨家林，窦薇．普洱红茶和普洱绿茶降低老年人血脂的对比观察．中国老年学杂志1997，6(17)：174
49．谢一辉，罗永明．茶色素胶囊中总多酚的紫外分光光度法测定．中国医药工业杂志，2001，32(5)：227-230
50．夏勇，徐彩菊．茶色素对实验性高脂血症大鼠血脂水平的影响浙江预防医学．2003，15 (7)：76-77
51．邵翔，杨浩，俞国华．茶提取物在心血管疾病中的应用．现代中西医结合杂志．2005，14(14)：1917-1919
52．李大祥，宛晓春．茶色素中茶黄素类的HPLC定量，茶叶科学2004，24(2)：124—128
53．郑青山，丁学庭，宋寅．茶多酚对大鼠实验性高血脂的影响．安徽医药．1997 71(2)
54．王汉生．茶多酚降血脂及抗氧化作用动物实验．华南农业大学
55．龚志华，仇灿红，肖文军．茶多酚降血脂研究进展．福建茶叶2002(4)：39-40
56．宋小鸽．茶多酚提取工艺及降脂作用的概述．黑龙江中医药，1998(6)：51-53
57．林智，吕海鹏，崔文锐．普洱茶的抗氧化酚类化学成分的研究．茶叶科学2006，26(2)：112～116
58．杨慧琴，赵斌，王江民．茶色素治疗高脂血症附150例疗效观察．邯郸医学高等专科学校学报，2000，13(6)：443-444
59．王路宏，钟永香．茶色素调节血脂代谢紊乱90例．中国新药与临床杂志，1999，11，18(6)：409—410
60. Helaine M. Alessio, Ann E. Hagerman. Consumption of green tea protects rats from exerciseinduced oxidative stress in kidney and liver. Nutrition Research 22 (2002): 1177-1188
61. J.J. Choo. Green tea reduces body fat accretion caused by high-fat diet in rats through β-adrenoceptor activation of thermogenesis in brown adipose tissue. Journal of Nutritional Biochemistry 11 (2003): 671-676
62. Ming-Hua Yang, Cheng-Hsin Wang. Green, oolong and black tea extracts modulate lipid mePEbolism in hyrlipidemia rats fed high-sucrose diet. Journal of Nutritional Biochemistry 12 (2001) 14-20
63. M. LODOVICI, C, CASALINI. Inhibition of 1, 2-dimethylhydrazine-induced oxidative DNA damage in rat colon mucosa by black tea complex polyphenols Food and Chemical Toxicology 38 (2000): 1085-1088
64. Rajesh Krishnan, Girish B. Maru Isolation and analyses of polymeric polypbenol fractions from black tea Food Chemistry 94 (2006) 331-340
65. Vasyl M. Sava, Swen-Ming Yang. Isolation and characterization of melanic pigments derived from tea and tea polypbenols.Food Chemistry 73(2001) 177-184
66. S. De Vos, R. De Schrijver. Lipid mePEbolism, intestinal fermenPEtion and mineral absorption in rats consuming black tea. Nutrition Research 23 (2003) 527-537
67. E. Skrzydlewska, J. Ostrowska, R. Farbiszewski. Protective effect of green tea against lipid roxidation in the rat liver, blood serum and the brain. Phytomedicine 9: 232-238, 2002
68. Nuria Martin-Carron, BSc, Isabel Goni, Ph.D. Reduction in serum toPEl and LDL cholesterol concentrations by a diePEry fiber and polyphenol-rich gra product in hyrcholesterolemic rats. Nutrition Research, Vol, 19, No.9, pp. 1371-1381, 1999
69. Scavenging Effects of Green Tea Polypbenols on the Oxygen Free Radicals Generated From Isolated Iscbemic Rerfused Rat Myocardium
70. Inoue M, Suzuki R, Koide T. Antioxidant, gallic acid. induces apoptosis in HL-60RG cells. Biocbem Biophys Res Commun, 1994, 204 (2): 898-904
71. Inouc M, Suzuki R, Sakaguchi N. Selective induction of cell detch in cancer cells by gallic acid. Biol Pharm Bull, 1995, 18 (11): 1526-1530
72. Inoue M, Sakaguchi N, lsuzugawa K. Role of reactive oxygen scies in gallic acid. induced apoptosis. Biol Pharm Bull, 2000, 23 (10): 1153-1157
73. Mukberjee G, Banerjee R. Biosynthesis of PEnnase and gallic acid from PEnnin rich substrates by Rhizopus oryzae and Asrgillus foetidus. Basic Microbio, 2004, 44(1): 42-8
74. Olmo T, lnoue M, Ogihara Y. Cytotoxic activity of gallic acid against liver mePEsPEsis of mastocytoma cells P815. Anticancer Res, 2001, 21 (6A): 3875-3880
75. YEN Gow-chin, CHEN Hui-yin. Antioxidant activity of tea extracts in relation to their antimuPEgenicity. J Agric Food chem., 1997, 45:30-34
76. Zhou ZH (周志宏), Yang CR (杨崇仁). Chemical constituents of crude green tea: the material of Pu'er tea in Yunnan. AcPE Bot Yunnan (云南植物研究), 2000, 22(3): 343-350