HPLC Profiling of Catechins and Purine Alkaloids in Leaves of Oolong and Green Tea Cultivars
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摘要: 为揭示适制乌龙茶和绿茶品种鲜叶的基本生化特性，对集中种植的10个乌龙茶品种和10个绿茶品种春季和秋季鲜叶（一芽二、三叶）进行儿茶素类和嘌呤碱高效液相色谱（HPLC）指纹图谱构建，并采用主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA）等方法进行可视化模式识别。结果表明，乌龙茶品种与绿茶品种鲜叶样存在良好的类群区分。乌龙茶品种鲜叶样的儿茶素类和嘌呤碱指纹图谱的模式分布较为离散，且存在明显的季节性差异。乌龙茶品种相较绿茶品种鲜叶样拥有更丰富的儿茶素类和嘌呤碱组分。咖啡碱（CAF）、可可碱（TB）、表儿茶素没食子酸酯（ECG）、儿茶素（C）、表没食子儿茶素（EGC）和1种未知化合物为鉴别乌龙茶品种与绿茶品种鲜叶样的主要差异标志物，而7种未知化合物和表没食子儿茶素（EGC）则可视为鉴别乌龙茶品种春季鲜叶样与秋季鲜叶样的重要差异标志物。该研究结果可为适制乌龙茶和绿茶品种的选育与鉴定提供参考依据。Abstract: The contents of catechins and purine alkaloids in two or three leaves with a bud from the spring and autumn shoots of 10 oolong and 10 green tea cultivars (Camellia sinensis) were determined by high performance liquid chromatography. A visual pattern recognition was performed with the aid of the principal components analysis, orthogonal partial least squares discriminant analysis and other statistical methods. There were distinctive differences on the profiles between the two categories of cultivars. The catechin and purine alkaloid fingerprints of oolong teas appeared more abundant and diverse than those of green teas. They also differed significantly by the season the leaves were plucked. Among the chemical constituents, caffeine, theobromine, epicatechin gallate, catechin, epigallocatechin, and an unknown component were particularly prominent. They could be used as the major markers for differentiating the two teas. In addition, 7 unknown components and epigallocatechin appeared suitable for distinguishing the spring shoots from the autumn shoots of oolong teas.
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Key words:
- catechins /
- purine alkaloids /
- pattern recognition /
- tea cultivars /
- fingerprint
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图 1 供试茶样儿茶素类和嘌呤碱HPLC指纹纵向叠加图谱
注：自上而下依次为OS01~OS10，OA01~OA10，GS01~GS10和GA01~GA10。
Figure 1. Catechin and purine alkaloid fingerprints of tea samples stacked in vertical pattern
图 2 40份供试茶样儿茶素类和嘌呤碱HPLC指纹图谱共有模式(高斯曲线)
Figure 2. Common pattern expressed by Gaussian equation on catechin and purine alkaloid fingerprints from 40 tea samples
图 4 正交偏最小二乘判别分析
注：A为得分图，A1为乌龙茶品种vs.绿茶品种，A2为春季鲜叶vs.秋季鲜叶(乌龙茶品种)；B为置换检验，B1为乌龙茶品种vs.绿茶品种，B2为春季鲜叶vs.秋季鲜叶(乌龙茶品种)。
Figure 4. Orthogonal partial least squares discriminant analysis
图 5 不同来源茶鲜叶儿茶素类和嘌呤碱指纹特征的差异分析
注：A为火山图，A1为乌龙茶品种vs.绿茶品种，A2为春季鲜叶vs.秋季鲜叶(乌龙茶品种)；B为S形曲线，B1为乌龙茶品种vs.绿茶品种，B2为春季鲜叶vs.秋季鲜叶(乌龙茶品种)。图A1中的红色三角形表示该成分在乌龙茶品种鲜叶样中含量较高，且与绿茶品种鲜叶样存在显著性水平差异(P < 0.05)；A2中的红色三角形表示该成分在乌龙茶品种春季鲜叶样中含量较高，而蓝色正方形表示该成分在乌龙茶品种秋季鲜叶样中含量较高，且二者存在显著性水平差异(P < 0.05)。B1的红色圆形为区分乌龙茶品种与绿茶品种鲜叶样的主要差异成分；图B2的红色圆形为鉴别乌龙茶品种春季鲜叶样与秋季鲜叶样的主要差异成分。
Figure 5. Discriminant components obtained from catechin and purine alkaloid fingerprints of various tea samples
表 1 茶样编码及其鲜叶来源
Table 1. Codes for teas and sample sources
样品编码 鲜叶来源 OS01 肉桂春季一芽二、三叶 OS02 水仙春季一芽二、三叶 OS03 大红袍春季一芽二、三叶 OS04 矮脚乌龙春季一芽二、三叶 OS05 铁观音春季一芽二、三叶 OS06 黄棪春季一芽二、三叶 OS07 梅占春季一芽二、三叶 OS08 白芽奇兰春季一芽二、三叶 OS09 金观音春季一芽二、三叶 OS10 黄观音春季一芽二、三叶 GS01 福鼎大白茶春季一芽二、三叶 GS02 福鼎大毫茶春季一芽二、三叶 GS03 福安大白茶春季一芽二、三叶 GS04 福云6号春季一芽二、三叶 GS05 福云7号春季一芽二、三叶 GS06 福云10号春季一芽二、三叶 GS07 霞浦春波绿春季一芽二、三叶 GS08 九龙大白茶春季一芽二、三叶 GS09 霞浦元宵茶春季一芽二、三叶 GS10 早春毫春季一芽二、三叶 OA01 肉桂秋季一芽二、三叶 OA02 水仙秋季一芽二、三叶 OA03 大红袍秋季一芽二、三叶 OA04 矮脚乌龙秋季一芽二、三叶 OA05 铁观音秋季一芽二、三叶 OA06 黄棪秋季一芽二、三叶 OA07 梅占秋季一芽二、三叶 OA08 白芽奇兰秋季一芽二、三叶 OA09 金观音秋季一芽二、三叶 OA10 黄观音秋季一芽二、三叶 GA01 福鼎大白茶秋季一芽二、三叶 GA02 福鼎大毫茶秋季一芽二、三叶 GA03 福安大白茶秋季一芽二、三叶 GA04 福云6号秋季一芽二、三叶 GA05 福云7号秋季一芽二、三叶 GA06 福云10号秋季一芽二、三叶 GA07 霞浦春波绿秋季一芽二、三叶 GA08 九龙大白茶秋季一芽二、三叶 GA09 霞浦元宵茶秋季一芽二、三叶 GA10 早春毫秋季一芽二、三叶 
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