摘要
A reliable and validated HPLC method was established for the assay of trypsin inhibitors（TI）and it was used in the investigation of the active TI components inMomordica cochinchinensis（Cucurbitaceae family）.The underlying principle of the assay is the measurement of the decrease in trypsin activity due to the presence of TI in analyzed samples,which was achieved by using HPLC separation and quantification of p-nitroanilide that was generated by tryptic hydrolysis of N-α-benzoyl-DL-arginine -4-nitroanilide.The results showed that the HPLC method had higher selectivity than conventional spectrophotometric assay.
本研究介绍了HPLC方法测定胰蛋白酶抑制剂活性及其在中药木鳖子胰蛋白酶抑制剂活性成分测定中的应用,由于分析样品中的胰蛋白酶抑制剂可以减低胰蛋白酶的活性,所以可以通过测定胰蛋白酶活性的下降来评价胰蛋白酶抑制剂的活性。测定原理是:胰蛋白酶能水解N-α-苯甲酰-DL-精氨酰-4-硝基苯胺盐酸盐生成产物4-硝基苯胺,采用HPLC方法对产物4-硝基苯胺进行分离和定量,最终测定样品中胰蛋白酶抑制剂的活性。该方法与一般的紫外分光光度法相比选择性更好。
基金
National Natural Science Foundation of China (Grant No 30873405)
National Drug Innovation Program(Grant No.2009ZX09301-011)
