《植物生理学报》 2012, 48(4): 413-417

纸层析-分光光度法检测茶氨酸

马雪泷¹, 邹鹏飞¹, 王荡强¹, 曹顺利², 房江育^1,3,*, 王昶³

1黄山学院生命与环境科学学院, 安徽黄山245041; 2安徽农业大学茶叶生物化学与生物技术农业部重点实验室, 合肥

通信作者：房江育；E-mail: fjy@hsu.edu.cn；Tel: 0559-2546631

摘　要：

为了满足普通实验室对茶中茶氨酸测定的需要, 研究了茶氨酸的纸层析-分光光度检测方法。结果表明, 用茚三酮-乙醇水溶液做展层剂, 对茶苗根、芽叶和茶叶的水浸提液进行纸层析, 能够有效地将茶氨酸与其它氨基酸分离, 紫色色斑清晰而均匀。用乙醇溶液洗脱色斑后用分光光度计在570 nm比色, 在20~70 μL茶氨酸溶液点样量范围内其含量与吸光度呈线性关系。本方法检出限为0.0057 mg·mL-1, 测定下限为0.0191 mg·mL-1, 平均回收率90.28%~115.38%, 平均相对标准差1.51%, 具有安全、药品种类少和操作步骤简单等特点。

关键词：茶; 茶氨酸; 茚三酮; 乙醇; 纸层析; 分光光度法

收稿：2011-11-21   修定：2012-03-14

资助：国家自然科学基金项目(30771755)、黄山学院引进博士启动资助项目(2012xkjq003)、黄山市科技孵化资金项目(2010F-03)和黄山学院大学生科研项目(2011xdkj043和2011xdkj056)。

Determination of Theanine in Tea (Camellia sinensis L.) by Paper Chromatography-Spectrophotometry

MA Xue-Long¹, ZOU Peng-Fei¹, WANG Dang-Qiang¹, CAO Shun-Li², FANG Jiang-Yu^1,3,*, WANG Chang³

¹College of Life and Environmental Sciences, Huangshan University, Huangshan, Anhui 245041, China; ²Key Laboratory of Tea Biochemistry and Biotechnology of Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, China; ³Huangshan Xiannong Bio-Technology Co., Ltd, Huangshan, Anhui 245900, China

Corresponding author: FANG Jiang-Yu; E-mail: fjy@hsu.edu.cn; Tel: 0559-2546631

Abstract:

To meet the needs of ordinary laboratory determination of theanine, paper chromatographicspectrophotometry was developed. The results showed that by paper chromatography, ninhydrin-ethanol water solution could effectively separate theanine from other amino acids extracted in the water from roots or shoots of tea seedling or from tea. The purple stains were clear and even. After the stains were eluted with ethanol solution, theanine content could be assayed with the spectrophotometer at 570 nm. The theanine content appeared a linera relationship with the absorption value when the solution was spotted on the paper at the range of 20-70 μL. The detection limit was 0.0057 mg·mL^-1 and the lowest detection limit was 0.0191 mg·mL^-1. The average recovery ranged from 90.28% to 115.38%, and the average relative standard deviation was 1.51%. The paper chromatography-spectrophotometry characterizes safety, fewer chemicals, and easy to operate.

Key words: tea; theanine; ninhydrin; ethanol; paper chromatography; spectrophotometry