纸层析法分离氨基酸实验条件的优化

Optimization of experimental conditions for the separation of amino acids by paper chromatograph

纸层析法分离氨基酸是生物专业本科实验必修课,但是实际实验过程中存在时间长、分离效果差、实验结果不稳定等现象.本研究分析了点样方式、扩展剂和展层方向等因素对氨基酸分离的影响.结果表明,用正丁醇∶甲酸∶水=15∶3∶2且溶入0.01%的茚三酮作为展层显色剂、用毛细管在环形滤纸中央环形区点样一次的水平纸层析方法可以在30~45 min将浓度0.2%混合氨基酸（赖氨酸、丝氨酸、丙氨酸、亮氨酸）明显分离开,取得最佳的分离显色效果.改进后的纸层析法比传统纸层析法具有速度更快、斑点更集中、分离效果明显、显色灵敏度高、节约药品等优点.

The paper chromatography separation of amino acids is a compulsory biological experiment for under- graduate. But some shortcomings, such as a long duration, poor separation effect and unstable experiment re- suits, exist in this experiment. This study conducted the influence factors of point mode, expanding agent and the developing direction on the separation of amino acids. The results show that, using capillary pipette point 0.2% amino acid mixture （including lys, ser, ala, leu） sample in the core of a horizontal circular paper in im- proved chromatography extension-color agents including n-butanol： formic acid： water （15： 3：2 ） integration O. Ol % ninhydrin could distinctly separate during 30 - 45 minute. The improved paper chromatography has the advantage of shorter separate time, more concentrated separate spots, higher color sensitivity, more obvious sepa- rate effect and less agent dose than traditional paper chromatography.