蝮蛇肉氨基酸的反相高效液相色谱分析

Determination of Amino Acids in Agkistrodon Halys (pallas) Protein Hydrolysate by Reversed Phase High Performance Liquid Chromatography

采用氨基酸与DABS-Cl试剂柱前衍生反应,生成DABS—氨基酸进入反相高效液相色谱(RP—HPLC),在436nm处分离18种氨基酸。DABS—氨基酸可达到0.01AUFS的灵敏范围;最低检测浓度为2.0μmol/L,批内变异系数<5%,重复性好。衍生反应在70～72℃保温15min,非常稳定。本法对蝮蛇(Agkistrodon halys(Pallas))肉水解物的色谱分析可分离出17种氨基酸。

A new and sensitive method for separation and quantification of amino acids by pre—column derivation with dabsyl chloride and reserved—phase HPLC are described. The use of the model SPD—6AM fixed wavelength(436nm) detector with a sensitivity limit of 0.01 AUFS permits amino acid analysis with stable baselines at levels. The Products of derivation were incubated at 70℃～72℃ for 15min and were very stable. In all instances the column was maintained at 40℃ and a flow—rate of 1ml/min was used. The minimum detectable concentration was 2.0μmol/L. The coefficient of variation for analysis within a day was less than 5%. The reproducibility of analysis was good. The method has been successfully applied to the determination to primary amino acids of Agkistrodon halys protein hydrolysate. Seventeen kinds of amino acids in the hydrolysate were successfully separated.