OPA柱前衍生法用于反相高效液相色谱分析氨基酸

Analysis of Amino Acids with Precolumn Derivatization of O-phthaldialdehyde and Reversed-phase High Performance Liquid Chromatography

本文采用邻苯二甲醛(OPA)柱前衍生的反相高效液相色谱法,以甲醇、0.1M柠檬酸缓冲液梯度洗脱,分析了细胞色素C酶解产物的氨基酸组成。结合原子吸收光谱,确证了铂在细胞色素C上的修饰作用位点。

The amino acids composition of peptides isolated from tryptic hydrolysate of cytochrome c modified by pt has been analysed by RP—HPLC with a methanoI gradient in O.1M aqueous sodium citrate.With the analysis of atomic absorption spectrum, the site of Pt binding with cytochrome c has been identified.The results have proved that the OPA precolumn derivatization is a rapid ultrasensitive and reproducible method which has a detection limit of less than 100 fmoles for analysis of amjno acids in peptide hydroIysates.