纸层析分离鉴定氨基酸实验改进.docVIP

纸层析分离鉴定氨基酸实验改进 　　摘要: 采用纸层析分离鉴定5种不同的氨基酸，对传统《基础生物化学实验》中的“氨基酸的分离鉴定――纸层析法”进行了改进，同时又与薄层层析分离鉴定氨基酸进行比较.结果表明，在进行氨基酸的分离鉴定时，改进的纸层析法比传统的纸层析法具有耗材少，时间短等优点；而薄层层析法又比纸层析法有更多的优越性，具有速度更快、斑点更集中、显色灵敏度更高等优点.? 　　关键词: 纸层析；薄层层析；氨基酸；?R??f值? 　　中图分类号:Q 503 文献标志码:A 文章编号:1672-8513(2011)03-0229-03 　　?? 　　Improvement of the Method on Amino-Acid Isolation? and Identification by Paper Chromatography 　　 　　YANG Li?1, YOU Li?1, YE Jin?xiu?1, YUAN Yan??1,2? 　　? 　　 (1.Key Laboratory of Ethnic Medicine Resource Chemistry, State Ethnic Affairs Commission Ministry of Education,?School of chemistry and Biotechnology, Yunnan University of Nationalities，Kunming 650031, China;? 2. Department of Life Science and Technology, Kunming University, Kunming 650031, China) 　　 　　Abstract: Five kinds of different amino acids were isolated and identified by paper chromatography. The traditional experiment of Isolation and Identification of Amino Acids Using Paper Chromatography in the book of Basic Biochemical Experiments was improved. And the isolation and identification of amino acids were compared by the paper and thin-layer chromatography. The results showed that the method of improvement paper chromatography had fewer wastes and shorter time than the traditional paper chromatography on the amino acids isolated and identified. And the thin-layer chromatography method has more advantages than the paper chromatography with faster and higher sensitivity.? 　　Key words: paper chromatography; thin-layer chromatography; amino acids; ?R??f value 　　?? 　　纸层析是利用样品各组分在固定相和流动相中分配系数的不同，从而使各组分达到分离的层析技术，广泛应用于氨基酸、蛋白质、染料、色素和中草药等物质的分离和鉴定［1-3］.在“基础生物化学实验”（第2版,高等教育出版社)，“氨基酸的分离鉴定――纸层析法”是学生实验的重要内容，但是按照该教材上的要求操作，比较烦琐，整个实验的时间很长［4］.因此，为了提高教学效果，我们对此实验作了改进，同时又与薄层层析作了比较，效果比较好.? 　　1 材料与方法? 　　1.1 材料? 　　 层析滤纸(新华一号)、薄层板、烧杯(250?mL)、表面皿、吹风机、点样毛细管、镊子、剪刀、尺和铅笔.? 　　1.2 试剂? 　　 0.5%的氨基酸溶液(赖氨酸、脯氨酸、缬氨酸、苯丙氨酸、亮氨酸)及其混合液（各组份浓度均为0.5%）；展层剂?V?（正丁醇）∶[KG-*2]?V?（冰醋酸）=4∶1）；显色剂（0.1%水合茚三酮正丁醇溶液）.? 　　1.3 方法? 　　1.3.1 传统纸层析法分离鉴定氨基酸的改进［5-6］? 　　1) 滤纸的准备.