柱前PITC衍生与反相高效液相色谱分析兔阑尾CaBP21的氨基酸含量

AMINO ACID ANALYSIS OF CaBP_21 IN RABBIT APPENDIX BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF PHENYLISOCARBAMYL AMINO ACIDS

A reverse-phase high-performance liquid chromatographic method was developed for the assty of phenylisothiocarbamyl amino acids on the Super Pac Cl8 column ( 125mm x 4.0mm, Sum). The derivatization method is simple, rapid, sensitive and specific for the analysis of primary and secondary amino acids in protein hydrolyzates. The liluid chromatographic detection is rapid (overall run within 18 mins ) , sensitive (lower than 40 pmole for accurate routine analysis), with good linearity and reproducibility (average relative standard deviation of retention time and peak area were 0.38% and 3.08% respectively). The amino acid composition of CaBP2I, a newly discovered calcium-binding protein from rabbit appendix B lymphocytes, was determined by this approach. The results showed it might be a novel calcium-binding protein with distinct amino acid composition, but its structure characteristics are similar to those of calmodulin.