不同产地、加工工艺及储存年限普洱茶化学成分和药理活性的比较研究

【作者】 金裕范；

【导师】 王文全；

【作者基本信息】 北京中医药大学 ， 中药学， 2012， 博士

【摘要】 普洱茶,是以云南特有的大叶茶[Camellia sinensis (Linn.) var. assamiea (Masters) Kitmaura]为原料,经特殊后发酵下艺加工而成的我国特种茶,口感滑嫩,具有降血脂、降血压、抑菌、助消化、解毒等多种保健功效,且具有浓厚的文化底蕴,深受消费者的青睐,被公认为绿色保健食品饮料。普洱茶主产云南,在贵州、广西、福建、广东和海南也有分布,以云南的临沧、普洱和西双版纳为原产地。随着普洱茶产业的兴旺,普洱茶的市场出现鱼目混珠的情况,不同产地、不同厂家、不同品牌的普洱茶质量参差不齐,严重影响了普洱茶市场和产业的健康发展。本论文在对现有普洱茶产地和市场进行充分调研的基础上,分别选择了红河、西双版纳、大理下关、思茅、临沧等五个不同产地、同一储存年限(3年)、同一加工工艺(熟)的普洱茶,储存时间为1年和3年的同一产地(红河)、同一加工工艺(生)的普洱茶,及同一产地(红河)、同一储存年限(3年)的生普洱茶和熟普洱茶,比较研究了其总黄酮、总多糖、代表性活性组分等化学组成的差异,同时比较了其抗氧化、抗真菌、降血脂等药理活性的差异,以确定不同产地、不同储存年限及不同加工工艺对普洱茶质量的影响。本论文的主要研究结果如下：1.随着普洱茶市场的扩张,普洱茶的产地也向原产地周边延伸。目前,红河、西双版纳、大理、思茅、临沧等地均有大面积的普洱茶产区。经过多年的发展,普洱茶已远销港、澳、台、东南亚及世界各国,并已形成了吉幸、大益、六大茶山等多个知名品牌。但目前普洱茶市场存在以次充好、以新充旧、假冒名牌、蒙骗顾客等多种问题,严重影响了普洱茶产业的健康发展,从理论角度阐明不同产地、不同储存年限及不同加工工艺对普洱茶质量的影响已迫在眉睫。2.选择红河、西双版纳、大理下关、思茅、临沧5个不同产地普洱茶,采用NaNO2-Al(NO3)3(?)去测定总黄酮含量,采用硫酸苯酚法测定总多糖含量,采用HPLC法测定没食子酸、咖啡因及(+)儿茶素含量,结果发现红河产普洱茶总多糖含量最低,为4.33%,而咖啡因和(+)儿茶素含量最高,分别为37.66 mg/g和0.49 mg/g；西双版纳产普洱茶总黄酮含量最高为5.94%；大理下关产普洱茶总多糖和没食子酸含量最高,分别为6.18%和15.12 mg/g；思茅产普洱茶总黄酮、咖啡因、没食子酸及(+)儿茶素含量均为最低,分别为2.93%、24.76 mg/g、2.78 mg/g和0.49 mg/g；临沧产普洱茶各项指标含量均居中。采用DPPH法比较其抗氧化活性,发现不同产地来源普洱茶存在一定差异,大理下关普洱茶>西双版纳普洱茶>临沧普洱茶>红河普洱茶>思茅普洱茶。采用MIC法比较抗真菌活性,发现不同产地的普洱茶存在较大的差异,以大理下关产普洱茶的抑菌活性最强,抑菌谱较广,思茅和临沧产普洱茶几无抗真菌活性,而红河和西双版纳产普洱茶抗真菌活性居中。采用高脂饲料饲喂小鼠高血脂模型,比较降血脂活性,发现其降血脂活性相当。虽然活性成分含量存在一定差异,但多种药理活性基本相当,本论文认为红河、西双版纳、大理下关、思茅、临沧产3年熟普洱茶质量相当。3.选择红河产储存1年的生普洱茶与储存3年的生普洱茶,采用NaNO2-Al(NO3)3法测定总黄酮含量,采用硫酸苯酚法测定总多糖含量,采用HPLC法测定没食子酸、咖啡因及(+)儿茶素含量,结果发现储存3年的普洱茶较储存1年的普洱茶除总黄酮含量显著升高外,总多糖、咖啡因和(+)几茶素含量均降低,而没食子酸含量两者相当。采用DPPH i去比较其抗氧化活性,发现储存1年的普洱茶DPPH清除能力比储存3年的普洱茶强。采用MIC法比较抗真菌活性,发现发现两者对白色念珠菌和烟草疫霉菌均无抑菌活性,对小麦赤霉菌的抑菌活性相当,对番茄灰霉疫病菌的抑菌活性储存1年的普洱茶略强于储存3年的普洱茶。采用高脂饲料饲喂小鼠高血脂模型,比较降血脂活性,发现两者降血脂活性无显著性差异,但储存3年的普洱茶在各项指标的改善方面均优于储存1年的普洱茶。在储存过程中,普洱茶的化学成分发生了较大的变化,随着储存年限的延长,总黄酮含量升高,总多糖及咖啡因、(+)儿茶素等活性成分含量降低,也有部分成分如没食子酸含量变化不明显。由于化学成分的变化,导致其药理活性也存在一定差异,随着储存年限的延长,抗氧化活性降低,而降血脂活性升高。4.选择红河产储存3年的生普洱茶与储存3年的熟普洱茶,采用NaNO2-Al(NO3)3法测定总黄酮含量,采用硫酸苯酚法测定总多糖含量,采用HPLC法测定没食子酸、咖啡因及(+)儿茶素含量,结果发现熟普洱茶与生普洱茶的主要活性成分及其配比存在显著性差异,经渥堆发酵后,熟普洱茶与生普洱茶相比,总多糖与没食子酸含量有一定程度的升高,而总黄酮与(+)儿茶素含量显著降低。采用DPPH法比较其抗氧化活性,发现生普洱茶DPPH清除能力显著强于比熟普洱茶。采用MIC法比较抗真菌活性,发现熟普洱茶的抑菌活性和抑菌谱均强于生普洱茶。采用高脂饲料饲喂小鼠高血脂模型,比较降血脂活性,发现两者降血脂活性无显著性差异,但熟普洱茶在各项指标的改善方面均优于生普洱茶。5.采用红外吸收光谱法,比较了5种不同产地来源、2种不同储存年限及2种不同加工工艺的普洱茶红外吸收光谱的差异,发现不同来源普洱茶红外指纹光谱存在一定差异,可通过特制区域和特制吸收的分析,判定普洱茶的产地、储存年限及加工工艺等信息,在普洱茶的质量监督方面有一定的利用价值,具体方法有待进一步研究。本论文的主要创新点如下：(1)采用产地调研和市场调研相结合的方式,准确获得了普洱茶产业发展的第一手资料；(2)采用多种化学分析手段和多种药理模型相结合的手段,对不同产地、不同储存年限、不同加工工艺对普洱茶化学成分和药理活性的影响进行了比较分析,系统评价了各因子对普洱茶质量的影响；(3)本论文的结果和数据,将为普洱茶产业的健康发展提供重要的试验依据。更多还原

【Abstract】 Pu’er tea, made of Camellia sinensis (Linn.) var.assamiea (Masters) Kitmaura, distributed only in Yunnan province, with special fermentation process, is a special tea of China. Pu’er tea, with a lubricous taste, has the functions of reducing blood fat, lowering blood pressure, antibacterial, helping digestion, and detoxification and various health care effects. In addition, it also has a strong cultural foundation, thus it becomes more and more popular among the consumers, and has been recognized as a green healthy drinks. It is mainly produced in Yunnan province. Besides, there is a distribution in Guizhou, Guangxi, Fujian, Guangdong and Hainan provinces. Lincang, Pu’er and Xishuangbanna in Yunnan province is thought to be the origin. With the development of Pu’er tea industry, there are more and more imitation and inferior issues in Pu’er tea markets. Quality differences exist between Pu’er tea from different areas, different manufacturers, different brands, which seriously affect the healthy development of Pu’er tea market and the industry. Based on a sufficient investigation of the production places and markets of Pu’er tea, those from five different area, including Honghe, Xishangbanna, Dalixiaguan, Simao and Lincang, those of different storage time, that is 1 year and 3 years, and those of different processing technology, that is crude and processed, were chosen in this thesis. To determine the effects of different origin, different storage time and different processing technology on quality of Pu’er tea, their contents of total flavonoids, total polysaccharides and other active components were investigated, and their antioxidant activity, antifungal acitivity and reducing blood lipid activity were tested and compared. The main results were as follows:1. Along with the expansion of Pu’er tea market, its production areas also extend to the peripheral areas of origins. So far, there has been a large distribution of Pu’er tea in Honghe, Xishuangbanna, Dali, Simao, Lincang and other places. After years of development, Pu’er tea has been exported to Hong Kong, Macao, Taiwan, Southeast Asia and other countries around the world. And a number of well-known brands has formed, such as Jixing, Dayi, six big tea hill, and so on. But at present, there are a variety of shoddy, fake and bad ones in the Pu’er tea market to cheat customers, which has seriously affected the healthy development of Pu’er tea industry. To elucidate the effects of different origin, different storage time and different processing technology on tea quality impact from a theoretical point of view is urgent. 2. Pu’er tea produce in the five different areas including Honghe, Xishuangbanna, Dalixiaguan, Simao and Lincang were chosen, NaNO2-Al(NO3)3 method was used to determine the content of total flavonoids, sulphuric acid phenol method was used to determine the content of total polysaccharides, HPLC method was used to determine the content of gallic acid, caffeine and (+) catechins, it was found that Pu’er tea from Honghe was lowest in polysaccharide content, and highest in the content of caffeine and (+) catechins, that was respectively 4.33%,37.66 mg/g and 0.49 mg/g. That from Xishuangbanna was highest in the content of total flavonoids, that was 5.94%. That from Dalixiaguan was highest in the content of total polysaccharides and galic acid, that was 6.18% and 15.12 mg/g, respectively. That from Simao was lowest in the contents of total flavonoids, caffeine, galic acid and (+) catechins, that was 2.93%,24.76 mg/g,2.78 mg/g and 0.49 mg/g, respectively. The content of all components of Pu’er tea from Lincang were in the middle. DPPH method was used to compare the antioxidant activity of different origin, and it was found there were certain differences among different origins. And the result was Dalixiaguan> Xishuangbanna> Lincang> Honghe> Simao. MIC method was used to compare the antifungal activity of different origin, and it was found there were large differences among different origins. That from Dalixiaguan had the strongest antifungal activity and broad antifungal spectrum, that from Simao and Lincang had rare antifungal activity, while that from Honghe and Xishuangbanna had the moderate antifungal activity. Hyperlipidemia mice model induced by high fat feed was used to compare their hypolipidemic activity, and it was found their hypolipidemic activity were equal. Although there were some differences among their contents of active components, all tested pharmacological activities of them were equal. It is proved there were no quality difference among the Pu’er tea from Honghe, Xishuangbanna, Dalixiaguan, Simao, and Lincang.3. Pu’er tea of 1 year storage time and 3 years storage time from Honghe were chosen, NaNO2-Al(NO3)3 method was used to determine the content of total flavonoids, sulphuric acid phenol method was used to determine the content of total polysaccharides, HPLC method was used to determine the content of gallic acid, caffeine and (+) catechins. It was found that Pu’er tea of 3-year storage was higher in the content of total flavonoids and lower in the contents of total polysaccharides, caffeine, and (+) catechins than that of 1-year storage, while their content of galic acid was equal. DPPH method was used to compare their antioxidant activity of different origin, and it was found that the antioxidant activity of Pu’er tea of 1-year storage was much stronger than that of 3-year storage. MIC method was used to compare the antifungal activity of different origin, and it was found that both of them had no effect on Candida albicans and Phytophthora nicotianae, they had equal effect on Wheat red mildew, and Pu’er tea of 1-year storage had stronger effect on Tomato gray mould blight fungus than that of 3-year storage. Hyperlipidemia mice model induced by high fat feed was used to compare their hypolipidemic activity, and it was found there were no significant difference between their hypolipidemic activity, although most the index of Pu’er tea of 3 year storage were better. The chemical components of Pu’er tea changed a lot during the storage. With the extension of storage time, the content of total flavonoids increased, the contents of total polysaccharides, caffeine and (+) catechin decreased, while the content of some components such as gallic acid did not change significantly. Due to the variation of chemical composition, there were certain difference among their pharmacological activities. The antioxidant activity decreased and hypolipidemic activity increased along with the increation of storage time.4. Both crude and processed Pu’er tea of 3 year storage time from Honghe were chosen, NaNO2-Al(NO3)3 method was used to determine the content of total flavonoids, sulphuric acid phenol method was used to determine the content of total polysaccharides, HPLC method was used to determine the content of gallic acid, caffeine and (+) catechins, it was found that there were significant difference in their active components and their proportion. After procession, the contents of total polysaccharides and galic acid increased at an extent, while the contents of total flavonoids and (+) catechins decreased significantly. DPPH method was used to compare their antioxidant activity, and it was found that the antioxidant activity of crude Pu’er tea was much stronger. MIC method was used to compare the antifungal activity, and it was found that the crude Pu’er tea had stronger antifungal activity and broad antifungal spectrum. Hyperlipidemia mice model induced by high fat feed was used to compare their hypolipidemic activity, and it was found there were no significant difference between their hypolipidemic activity, however, most the index of processed Pu’er tea were better. 5. Infrared absorption spectrometry was used to compare the infrared absorption spectral differences of Pu’er tea of 5 different origin,2 different storage times and 2 different processed technologies, it was found that there were certain difference in the IR fingerprint spectrum of Pu’er tea of different sources. Producing area, storage life and process technology could be identified through the special region and special absorption in the IR fingerprint spectrum. IR fingerprint spectrum has a certain value in the quality supervision of Pu’er tea, and further study is needed for the establishment of the specific method.The main innovations of this thesis are as follows:(1) the first hand information of the development of Pu’er tea industry was achieved through the survey of the origin and market of Pu’er tea. (2) A variety of chemical analysis methods and multiple pharmacological models were used to compare the chemical constituents and pharmacological activities of Pu’er tea of different origins, different storage years, different processing technology, and the effect of each factor on the quality of Pu’er tea was system evaluated. (3) The results and data of this study provided important experimental basis for the health development of Pu’er tea industry.更多还原