Factors Influencing the Effects of Theabrownin in Pu-erh Tea on Scavenging DPPH Radicals

[Objective] This study aimed to explore factors influencing the effects of theabrownin in Pu-erh tea on scavenging DPPH radicals. [Method] Taking Pu-erh tea as main research object, theabrownin in Pu-erh tea from tea piles with different times of mixture, stored for different years and processed by both the old and new techniques were extracted and isolated. And DPPH method was used to study the effects of theabrownin solution and tea liquor on scavenging radicals and their antioxidant activity. [Result] The result showed that the theabrownin content in Pu-erh tea from tea piles with one to four times of mixture revealed a first declining and subsequently rising trend. Under different pH conditions, the rates of DPPH radical scavenging of 1 mg/ml theabrownin solution made from tea piles with different times of mixture decreased gradually with the increase of pH, and all the rates were lower than that of the control vitamin C solution; the rates of DPPH radical scavenging of theabrownin from tea samples stored for different years and produced by different techniques dropped slowly within pH range 3.0-6.0; when pH was higher than 6.0, the rates of DPPH radical scavenging of theabrownin from tea samples produced by both new and traditional techniques in 2001 declined obviously, while the declining trend was not obvious in either the black tea or the Pu-erh tea produced by new technique in 2005. Under different theabrownin concentrations, the rates of DPPH radical scavenging of theabrownin solution made from tea piles with different times of mixture ascended as the concentration increased and the highest rate occurred in tea pile with four times of mixture, reaching up to 46.56%±0.88%; for tea samples stored for different years and produced by different techniques, the rates of DPPH radical scavenging of theabrownin rose gradually with the increase of the theabrownin concentration; the rate of radical scavenging of theabrownin in the black tea ascended obviously and that in Pu-erh tea was decreasing within the concentration range of 0.04 -0.06 mg/ml. At pH6.0, the DPPH radical scavenging ability of theabrownin in 0.1 mg/ml theabrownin solution and tea liquor containing 0.1mg/ml theabrownin from tea piles with different times of mixture ascended with the time of mixture; solution made from tea samples produced by different techniques in different years, the DPPH radical scavenging ability of theabrownin solution and tea liquor followed： Pu-erh tea produced by the new technique in 2001 ﹥ Pu-erh tea produced by the old technique in 2001﹥ Pu-erh tea produced by the new technique in 2005. And the DPPH radical scavenging ability of theabrownin in tea liquor containing 0.1 mg/ml theabrownin was higher than that in 0.1 mg/ml theabrownin Solution made from tea samples either with different times of mixture or produced by different techniques in different years. The above results indicated that many factor influenced the effects of theabrownin on eliminating DPPH radicals, including the time of fermentation, fermentation technique, years of storage, concentration and pH all had great influence on the effects of theabrownin on eliminating radicals. To ensure a good effect on scavenging radicals, it is better to prepare theabrownin at pH6.0 with concentration between 0.08 and 0.1 mg/ml for drinking. [Conclusion] Results of this study provide a basis for developing functional products of theabrownin from Pu-erh tea in the future.

Detection of composition of functional component theabrownins in Pu-erh tea by degradation method

Habitual tea consumption can reduce the risk of ASCVD morbidity and mortality and all-cause mortality in China,and long-term adherence to this habit can provide greater protection.Pu-erh tea is one of the representatives of post-fermented teas,and the polymers in Pu-erh tea have anti-cancer and anti-hyperlipidemic activities,but their chemical composition is still unknown.In this study,the chemical composition of Pu-erh tea was investigated in detail and eight(1–8)known compounds were isolated.The polymers in Pu-erh tea were also degraded by thiol degradation,and the fragments of quercetin and(–)-epicatechin-3-O-gallate were found in the thiol degradation products of Pu-erh tea polymers by UPLC observation and separation and purification.This is the first time that this fragment was obtained in Pu-erh tea polymer.

Chemical Constituents of Pu-erh Tea and Its Inhibition Effect on α-amylase in vitro

[Objective]The aim was to clarify the chemical substance basis of hypoglycemic and lipid-lowering effects of Pu-erh Tea. [Method]Pu-erh Tea was extracted with 95% ethanol,followed by petroleum ether,chloroform,ethyl acetate and n-butanol extraction,after the further purification and through the NKA-9 macroporous resin and many times of Sephadex column chromatography,two compounds were isolated,in the same time,the effect of Uracil and Gallic acid on α-amylase was studied. [Result]The Uracil and Gallic acid were isolated and identified respectively from Pu-erh Tea and the Uracil was firstly isolated from Pu-erh Tea; Gallic acid had strong inhibition on α-amylase. [Conclusion]It could provide some theories on the hypoglycemic and lipid-lowering effects of Pu-erh Tea.

Study on Pu-erh Tea Methanol Extracts on Antibacterial Activity of Several Plant Pathogens

[Objective] The aim was to study Pu-erh tea methanol extracts on antibacterial activity of several plant pathogens and provide reference for application of Pu-erh tea in agriculture disease prevention and control.[Method]The inhibitive effect of Pu-erh tea methanol extracts on several plant pathogens was studied by adopting mycelium growth rate method.[Result] The results showed that Pu-erh tea methanol extracts had inhibitive effects on Thanatephorus cucomeris(Frank)Donk,Fusarium oxysporum f.sp.niveum,Fusa...

Analysis of α-amylase Inhibitor Content Change in Pu-erh Tea During Pile-fermentation Process

The study was to investigate the changes of α-amylase inhibitor content in Pu-erh tea during pile-fermentation process. Pu-erh tea samples from two regions of Shuangjiang County and Jinggu Dai and Yi Autonomous County of Yunnan Province at various fermentation stages were used as experimental materials to investigate the effect of different fermentation stages on the inhibitory effect to α-amylase; and the change law of the inhibitory effect of c-amylase inhibitor during processing was meanwhile studied by determining the contents of tea polyphenol and amino acid. The results showed that crude meterial of Pu-erh tea presented strong inhibitory effect to α-amylase; this inhibitory effect assumed a de： creasing trend to the minimum at the middle stage of fermentation, whereafter it increased to some extent. Made tea also showed a strong inhibitory effect to α-amylase. During whole processing period, contents of tea polyphenol and amino acid generally assumed a remarkably decreasing trend. Our results provided references for further isolating co-amylase inhibitor from Pu-erh tea and discussing the mechanism of its health care function.

Analysis of Monosaccharide Composition of Pu-erh Tea Polysaccaride by Pre-column Derivatization HPLC

[Objective] This study aimed to establish a pre-column derivatization HPLC method for the identification and analysis of monosaccharide composition of Pu-erh tea polysaccharide. [Method] Pu-erh tea polysaccharide was extracted using the wa- ter extraction method, further isolated and purified by DEAE cellulose-52 columns. The obtained tea polysaccharide and four components TPS1, TPS2, TPS3 and TPS, were first derived by 1-phenyl-3-methyl-5-pyrazolone （PMP）, and then the PMP derivatives of monosaccharide were analyzed by high performance liquid chromatog- raphy （HPLC）. [Result] Pu-erh tea polysaccharide contained eight kinds of monosac- chaddes （mannose, rhamnose, glucuronic acid, galacturonic acid, grucose, galactose, arabinose, fucose）, without xylose; so it was the same with TPS1; each of TPS2, TPS3 and TPS4 contained seven monosaccharides, while no fucose. [Conclusion] This method is simplified and rapid, which can be used to determine the monosac- charide composition of Pu-erh tea polysaccharide and monosaccharide content.

Inhibitory Effects of Pu-erh Tea Leaves on Mouse Type IV Allergy

Pu-erh tea is one of post fermented tea manufactured through oxidative maturing by microorganisms. We investigated the preventive effects of pu-erh tea extract on oxazolone-induced type IV allergy in male (ICR) mice. Oral administration of 50 mg/kg water extract of pu-erh tea leaves resulted in significant preventive effects against mouse type IV allergy. The hydrophilic Theabrownin-like fraction (TBW-ND), with a high molecular weight of approximately 12,000 or higher, was prepared from pu-erh tea leaves by a solvent-extraction method, followed by Diaion HP-20 column chro-matography and dialysis. Oral and percutaneous administrations of TBW-ND at doses of 18.7 mg/kg and 0.037 mg/ear also resulted in significant preventive effects, which included preventing increases in levels of the proinflam-matory cytokine interleukin-12. The anti-allergic ingredients in TBW-ND were predicted to be highly complex compounds containing of polyphenols, polysaccharides and/or proteins. The results suggest pu-erh tea leaves could be a beneficial food for protecting against delayed-type allergy.

Study on Extraction Technology of Theabrownine from Liubao Tea and Its Antioxidant Activity

[ Objective] This study aimed to provide theoretical reference for the extraction of theabrownine from Liubao tea, determination of the biological activity, and development and utilization of theabrownine. [Method] Orthogonal experimental design was adopted to investigate the effects of solid-liquid ratio, extraction temperature, extraction duration, and extraction frequency on the yield of theabrownine extracted from Liubao tea. The extraction technology was optimized, and the antioxidant activity of theabrownine was analyzed. [ Result] The results showed that extraction frequency and extraction temperature had significant effects on the yield of theabrownine extracted from Liubao tea, while solid-liquid ratio and extraction duration had no significant effect. The optimal conditions for extracting theabrownine from Liubao tea were determined as follows ： solid-liquid ratio was 1 ： 30, extraction temperature was 100℃, extraction duration was 80 min, and extraction frequency was 4 times. Under these conditions, the yield of theabrownine reached 22.92%. Results of antioxidant activity determination showed that hydroxyl radical-scavenging activity of theabrownine was as high as 74%, and the scavenging effects was significantly higher than that of Vitamin C under low concentrations; nitroso-scavenging activity of theabrownine reached 73%, with no significant difference from that of Vitamin C; superoxide anion free radical-scavenging activity of theabrownine could be above 60%, which was lower than that of Vitamin C. [Condusion] Liubao tea contains abundant theabrownine that has relatively high antioxidant activity with good prospects in antioxidant application, which lays solid foundation for in-depth development of Liubao tea.

Characterization of aroma compounds of Pu-erh ripen tea using solvent assisted flavor evaporation coupled with gas chromatography-mass spectrometry and gas chromatography-olfactometry

Pu-erh ripen tea becomes popular worldwide owing to its unique taste and aroma.In the present study,volatile compounds from Pu-erh ripen tea were extracted using a solvent assisted flavor evaporation(SAFE)and analyzed with a gas chromatography-mass spectrometry(GC-MS)and a gas chromatography olfactometry(GC-O).Results demonstrated that 58 major volatile compounds and 24 aroma active substances of Pu-erh ripen tea were identified by GC-MS and GC-O analysis.Among volatile compounds identified,methoxyphenyl compounds were considered as the dominate compounds in the aroma of Pu-erh ripen tea.Further investigation showed that 6 methoxybenzenes were responsible for the aging aroma in Pu-erh ripen tea;α-ionone,3,4-dimethoxytoluene,1,3-dimethoxybenzene,etc.were responsible for the wood-like aroma.Linalool,α-terpineol,α-ionone,β-ionone,and benzeneacetaldehyde were responsible for the floral odor.Compared the odor detection threshold(ODT)among the 8 methoxyphenyl compounds in water,1,2,3-trimethoxybenzene showed the highest ODT,followed by 3,4-dimethoxyphenol.1,3-dimethoxybenzene(105μg/L)and 1,2,3-trimethoxy-5-methylbenzene(100μg/L)were present at relatively low odor threshold values.The result indicated that methoxyphenyl compounds play a vital role in the unique flavor of Pu-erh ripen tea.

Reducing oxidative stress and hepatoprotective effect of water extracts from Pu-erh tea on rats with high-fat diet

Reducing oxidative stress and hepatoprotective effect of Pu-erh tea water extracts on rats fed with high-fat diet were researched for explaining health care of Pu-erh tea.Fifty SD rats were divided into five groups.The body weight was measured once a day.The malondialdehyde(MDA)and glucose(Glu)levels and the activities of alanine aminotransferase(ALT),aspartate aminotransferase(AST),nitric oxide synthase(NOS),and pyruvate kinase(PK)in serum were determined.Furthermore,the hepatic glycogen level(HGL)and the activities of hepatic total superoxide dismutase(T-SOD),catalase(CAT),and glutathione peroxidase(GSH-Px)were also measured after continuous administration for 12 weeks.The result demonstrated that Pu-erh extract caused the decreases in body weight,fat index,MDA and NOS levels,and the increases in hepatic T-SOD,CAT and GSH-Px activities,indicating that the extract may be due to inhibiting the increases of body weight and fat index,reducing oxidant stress state and inhibiting lipid peroxidation,thus decreasing the activities of ALT and AST,and protecting the liver in rat.Meanwhile,the extracts could increase the production of hepatic glycogen and the activity of PK,and reduce glucose level,protecting the liver from the diseases associated with type II diabetes.

Pu-erh tea: A review of a healthful brew

Pu-erh tea has been used for thousands of years to treat metabolic diseases.Recognized in Shen Nong's Herbal Classic,a compendium kept by the first traditional Chinese practitioners,it is still highly valued for its hypocholesterolemic and hypolipidemic effects.This review reports the processing and bioactive components of pu-erh tea.Recent human and animal studies of pu-erh tea and its potential therapeutic mechanisms have also been summarized.The interaction of liver and gut microbiome regulates the puerh tea biotransformation and endogenous metabolism,and thus contributes to the health benefits.

Anti-Obesity, Anti-Atherosclerotic and Anti-Oxidant Effects of Pu-Erh Tea on a High Fat Diet-Induced Obese Rat Model

Pu-erh tea, a traditional Chinese beverage, has been believed to have many benefits to human health and without side effects. In this study, we systematically analyzed the main active components of Pu-erh tea and investigated its anti-obesity, anti-atherosclerotic and anti-oxidant effects using an obese rat model. Obesity was induced by feeding a high-fat diet and subsequently the experimental obese mice were fed with high-fat diet supplemented with low (2.5%), medium (5%) or high (7.5%) doses of Pu-erh tea powder for 6 weeks respectively. As result, the body weight gain of the rats was decreased by medium and high doses of Pu-erh tea treatments. Furthermore, the levels of serum total cholesterol (TC), triglyceride (TG) and atherosclerosis index (AI) were significantly lowered by Pu-erh tea compared to the control group. Conversely, high density lipoproteincholesterol (HDL-C) level of the rats was significantly elevated by Pu-erh tea treatments. In addition, Pu-erh tea treatments increased the activities of anti-oxidative enzymes such as superoxide dismutase (SOD) and glutathione peroxides (GSH-Px), whereas reduced the level of lipid peroxidation product malondialdehyde (MDA) in obese rats. Collectively, our find-ings revealed that Pu-erh tea exerts comprehensive benefits in anti-obesity, anti-atherosclerotic and anti-oxidant effects, therefore can be used as a promising functional food in obesity management.

氧化型粗茶褐素制备工艺优化及其抗氧化活性分析

以普洱生茶为原料制备氧化型粗茶褐素。以茶褐素含量为指标,在单因素试验基础上通过正交试验优化氧化型粗茶褐素的制备工艺,并与熟茶水提物对比分析其抗氧化活性。结果表明:氧化型粗茶褐素的最佳制备工艺为料液比1∶20(g/mL)、pH 8.5、通气时间14 h,在此条件下制得的氧化型粗茶褐素中茶褐素含量为77.12%。抗氧化试验结果表明,氧化型粗茶褐素与熟茶水提物的抗氧化能力相近,均具有较好的抗氧化活性。

菌方普洱熟茶水提物和茶褐素的组成及其降脂作用

研究菌方普洱熟茶水提物和茶褐素的组成及对大鼠肥胖的调节作用。基于居里点热裂解气相色谱-质谱联用技术及高效液相色谱技术,分析菌方普洱熟茶水提物和茶褐素的组成成分。设置正常对照组、高脂模型组、菌方普洱熟茶水提物组和菌方茶褐素组,连续灌胃16周,分析SD大鼠生长指标、血清生化指标、氧化应激及炎症相关指标,并对肝脏组织HE和油红O染色切片进行观察。结果表明:菌方普洱熟茶水提物与茶褐素粗品中茶褐素、蛋白质和多糖占较大比重;没食子酸、表儿茶素和表没食子儿茶素没食子酸酯等儿茶素及衍生物类化合物含量存在明显差异(P<0.05),菌方普洱熟茶水提物中生物碱类咖啡因和可可碱含量显著高于茶褐素粗品(P<0.05);在不同居里点热裂解温度下,菌方普洱熟茶水提物与茶褐素的裂解产物分别有7种(380℃)和12种共同成分(600℃),两者在不同温度下的裂解产物中,酚类种类最多,含量最高的化合物是咖啡因。与高脂模型组相比,菌方普洱熟茶水提物组和茶褐素组体质量、Lee’s指数显著减轻(P<0.05),血清甘油三酯、血清空腹血糖、肝脏甘油三酯和胆固醇水平显著降低(P<0.05),脂肪累积减少,血清超氧化物歧化酶、谷胱甘肽过氧化物酶、大鼠核因子E2相关因子2、白细胞介素-6、白细胞介素-1β、肿瘤坏死因子-α水平显著降低(P<0.05),表明抗氧化活性提高和炎症因子浓度降低,并且肝脏切片显示脂肪变性减轻、脂肪细胞数量和脂滴聚集明显减少。综上,菌方普洱熟茶水提物与其茶褐素组成成分存在很大区别,但两者在减肥降脂方面均呈现良好效果,研究可为茶叶资源的深加工和保健功能产品的开发提供理论依据。

基于pH高效定量判断普洱茶发酵程度的研究

目的 探索一种方便、快捷的方法来判断普洱茶的发酵程度。方法 使用目标容量的发酵罐,对普洱茶原料进行接种发酵,收集多组茶叶pH,没食子酸(gallicacid,GA)和茶褐素(theabrownin,TB)含量的动态变化数据,基于上述数据分别进行线性拟合并组合,得到TB含量随茶叶pH变化的回归模型。结果 在普洱茶发酵过程的特定阶段,分别建立了TB-GA、GA-茶叶pH的一元线性回归模型;然后将两个一元线性回归模型线性组合,最终建立了TB含量与茶叶pH的回归模型,只需根据测得的茶叶pH,即可快速对普洱茶发酵体系中TB含量进行预测,从而判定普洱茶发酵程度。结论 基于pH定量高效判断普洱茶发酵程度,保证了发酵过程参数调控的时效性和准确性,使产品稳定生产。

渥堆发酵过程中藏茶化学成分的变化

目的:阐明藏茶渥堆过程主要化学成分的变化规律。方法:以不同发酵阶段藏茶样为原料,研究发酵过程中藏茶水提物中可溶性固形物、可溶性糖、总多酚、总黄酮含量及水提物体外抗氧化活性的变化,并应用超高效液相色谱仪检测水提物中茶碱、咖啡碱、没食子酸和8种儿茶素类单体的含量,应用酶联免疫法测定水提物中茶黄素、茶红素和茶褐素的含量。结果:藏茶发酵过程中可溶性固形物含量先降低后略有回升,可溶性糖、总多酚、总黄酮含量逐渐降低,抗氧化活性逐渐减弱,咖啡碱呈波动状态且变化较小,没食子酸与儿茶素含量先上升再略有下降,没食子儿茶素没食子酸酯与表儿茶素含量先升高后降低,没食子儿茶素、表没食子儿茶素、表儿茶素没食子酸酯和表没食子儿茶素没食子酸酯含量均呈持续下降的状态,其中表没食子儿茶素没食子酸酯在黑毛茶中含量为39.69 mg/g,在成品茶中降至1.36 mg/g;茶红素含量先增加后降低,茶黄素含量变化很小,茶褐素含量逐渐升高(黑毛茶中3.12 g/100 g,成品茶中7.46 g/100 g)。结论:藏茶渥堆发酵过程中,可溶性糖、总多酚、总黄酮等含量降低,抗氧化活性减弱,茶碱、咖啡碱、儿茶素类单体等活性成分含量的变化趋势不一致,茶褐色含量逐渐升高。

英红九号发酵叶中茶褐素近红外定量模型的优化与验证

为提高近红外光谱分析方法快速实现对测定红茶在制品中的茶褐素的定量模型精度无损、快速检测,该试验利用近红外光谱技术对以英红九号发酵叶中茶褐素进行采集、提取和分析的检测为例,对其近红外定量检测模型的构建与优化进行了研究。首先,采用规范化处理(Normalize)、基线校正(Baseline)、S-G一阶导数(Savitzky-Golay,1^(st)S-G)、S-G二阶导数(2^(nd) S-G)、标准正态变量变换(Standard Normal Variate Transform,SNV)五种预处理方法对原始光谱进行预处理分析。然后,采用效果最好的一阶导数预处理方法进行波长特征提取,分别使用间隔偏最小二乘算法(IntervalPartialLeast Square,iPLS)、竞争自适应加权算法(Competitive Adaptive Reweighted Sampling,CARS)、变量迭代空间收缩方法(the Variable Iterative Space Shrinkage Approach,VISSA)提取波长特征。最后,使用偏最小二乘回归(PartialLeastSquare,PLS)预测模型进行回归建模。研究结果表明:使用一阶导数进行预处理、同时使用CARS方法建立的1^(st)-CARS-PLS模型效果特征更显著,特征值数量为53个。研究表明,该试验采用的模型方法能够快速、无损地检测英红九号发酵叶中的茶褐素含量。

茶树修剪老叶成分分析及利用塔宾曲霉制备茶褐素的条件优化

以大叶晒青茶(正常和粗老)为对照,分析冬季茶树修剪老叶的理化性质,并以修剪老叶水提物为原料,接种塔宾曲霉,纯菌株液态发酵制备茶褐素。结果表明,正常和粗老大叶晒青茶的营养成分差异小,冬季茶树修剪老叶的水溶性多糖、氨基酸、蛋白质、总多酚、总黄酮、儿茶素、茶黄素、茶红素和茶褐素含量均显著低于正常和粗老大叶晒青茶(P<0.05),而水溶性总糖显著高于正常和粗老茶(P<0.05)。塔宾曲霉发酵茶树修剪老叶水提物合成茶褐素的最优条件为:水提物质量浓度10 g/L,发酵温度40℃,摇床转速150 r/min。在最优条件下,茶褐素的含量为5.78 g/L,生成速率0.026 g/L·h,底物转化率3.922 g/g。本研究为提升冬季茶树修剪老叶的综合利用,降低茶褐素的生产成本提供依据。

普洱茶茶褐素提取工艺及理化性质的初步研究

研究了从普洱茶中提取茶褐素的工艺,并对所得茶褐素进行了红外光谱和主要理化性质分析。采用正交试验设计方法考察料液比、提取次数、提取温度、提取时间对茶褐素产率的影响。结果表明,提取次数对茶褐素产率的影响达到显著水平,料液比、提取温度和提取时间影响不显著。优化工艺条件为:料液比1∶10(g∶mL),提取次数3次,提取温度83℃,提取时间20 min,在此条件下产率达16.86%。茶褐素的总酸性基团为4.91 mmol/g,其中酚羟基为4.33 mmol/g,羧基为0.58 mmol/g。红外光谱和理化性质分析表明,茶褐素主要是由酚类和羧酸类物质构成,并含有蛋白质、多糖等化合物。

普洱茶茶褐素类主要组分特征及光谱学性质研究

利用不同分子截留量的透析袋可有效将茶褐素类物质分离。随分子量增大,茶黄素(TF)、茶红素(TR)、茶褐素(TB)、茶多糖(TPS)含量逐渐增加,分子量大于25 000Da的样品中TB含量最高。羧基和羟基含量随分子量增大而增加,特别是总羧基含量在分子量大于25 000Da的样品中增幅最大。AFM显示,不同分子量茶褐素粒子形貌并不均一,单分子呈岛屿状态或颗粒状聚集物结构。当粒子聚集较多时,呈线状链接且有较多分支或形成网状结构。CP-MAS NMR表明,在3 500～25 000Da的茶褐素属含有多苯环的苯多酚类高聚物,结合多糖、蛋白质残基,富含羧基、羟基、甲基等基团,具有酚类物质特性。其酸未水解物经CP-MAS NMR分析表明为多苯环高聚物,CP-GC/MS共鉴定出16种可能存在的化合物。