柱前衍生化-HPLC测定马氏珠母贝肉中氨基酸的含量

Detection and quantification of amino acids of Pinctada martensii by reversed-phase High Performance Liquid Chromatography

以邻苯二甲醛-9-芴甲基氯甲酸酯为衍生化试剂，对氨基酸进行柱前衍生，建立了35℃下，醋酸钠缓冲液、甲醇及乙腈为流动相，用EclipseAAA氨基酸分析柱和二极管阵列检测器的氨基酸色谱分析方法，18种氨基酸在36min内获得了较好的分离，在一定浓度范围内，氨基酸的峰面积与浓度的线性相关系数在0．9906～0．9998之间，相对标准偏差为1．96％～5．45％（n=5）。并测定了马氏珠母贝肉蛋白中氨基酸的组成和含量。结果表明，马氏珠母贝肉蛋白中含有至少18种氨基酸，其中8种必须氨基酸总含量达39％以上，是一种优质的功能食品原料。

A reversed-phase high-performance liquid chromatographic method for the determination of amino acids is described. It involves pre-column derivatization with o-phthalaldehyde and 9-fluorenylmethylchloroformate （FOMC） for the mixture of 18 kinds of amino acids. HPLC separation of the derivatives was performed using an Eclipse AAA column with NaAc buffer, methanol and aeetonitrile mobile phase system with DAD detector and at 35℃ in 36 min. The response was linear for all amino acids with a correlation coefficient of 0. 9906 - 0. 9998, Method precisions （ n = 5 ） for the different amino acids were between 2.02% -5.45% （relative standard deviation, RSD）. Using this method, 18 kinds of amino acids in protein of Pinctada martensii were determined. The results showed that there were at least 18 kinds of amino acids in proteins of Pinctada martensii in which the total quantity of 8 essential amino acids was about 39%. Pinctada martensii is a good functional food material.