降纤酶肽图分析

Peptide mapping analysis of defibrase

目的采用反相高效液相色谱法测定尖吻蝮蛇降纤酶和白眉蝮蛇降纤酶肽图。方法将降纤酶样品用胰蛋白酶水解,酶解所得的肽段通过反相高效液相色谱法进行分析,采用C18色谱柱,以0.1%三氟乙酸为A相,乙腈-水-三氟乙酸(90∶10∶0.1)为B相进行梯度洗脱得到降纤酶肽图。结果降纤酶各个肽段都能得到很好的分离,白眉蝮蛇降纤酶和尖吻蝮蛇降纤酶肽图存在显著差异。结论本法简便、快速,可应用于不同蛇种来源降纤酶的鉴别,以减少临床不良反应的发生。

Purpose To develop a RP-HPLC method for peptide mapping analysis of Agkistrodon acutus defibrase and Gloydius ussuriensis defibrase. Methods The peptide mapping was obtained by tryp- sin hydrolysis and RP-HPLC detection. The analysis was performed on a Cls column and the mobile phase consisted of 0.1% trifluotroacetie and acetonitrile-water-trifluotroacetic （90：10：0.1 ） with gradient elu- tion. Results The digested fragments were well separated and there was a significant difference between Gloydius ussuriensis defibrase and Agkistrodon acutus defibrase. Conclusion The method is simple and rapid for identification of defibrase from different sources to reduce the occurrence of adverse reactions.