UHPLC-MS/MS同时测定鳖甲煎丸中鳖甲胶和阿胶的特征肽含量

Simultaneous Determination of Marker Peptides of Trionycis-shell Glue and Asini Corii Colla in Biejiajian Pills by UHPLC-MS/MS

目的 建立同时测定鳖甲煎丸中鳖甲胶和阿胶特征肽含量的分析方法,为鳖甲煎丸的质量控制提供技术支持。方法采用胰蛋白酶对鳖甲煎丸提取溶液进行酶解,利用超高效液相色谱-串联质谱法(UHPLC-MS/MS),在电喷雾电离(ESI)正离子模式下采用多反应监测(multiple reaction monitoring,MRM),用外标法对鳖甲胶和阿胶的特征多肽进行含量测定。结果 方法学研究表明,该方法具有较好的专属性、精密度、稳定性、重复性。鳖甲特征肽A、驴源多肽A1、驴源多肽A2在线性范围内线性关系良好,相关系数均>0.999。3种特征多肽的平均回收率为97.08%~105.10%。结论 所建立的方法可用于鳖甲煎丸中鳖甲胶和阿胶的质量控制。鳖甲煎丸中鳖甲胶、阿胶的特征多肽含量低于拟定限度,应引起有关单位的重视。

OBJECTIVE To establish an analytical method for simultaneous determination of marker peptides contents of trionycis-shell glue and Asini Corii Colla in Bejiajian pills,and provide technical support for quality control of Bejiajian pills.METHODS Biejiajian pills was enzymolized by trypsin,the detection was performed by UHPLC-MS/MS in the ESI positive mode and MRM mode.The content of marker peptides in trionycis-shell glue and Asini Corii Colla was determined by external standard method.RESULTS The methodological evaluation showed that the method had good specificity,precision,stability,repeatability.Good linear relationships of trionychid marker peptide A,donkey polypeptide A1 and donkey polypeptide A2 were obtained and the correlation coefficients(r) were>0.999.The average recoveries of the 3 marker peptides were in the range of 97.08%-105.10%.CONCLUSION The established method can be used for the quality control of trionycis-shell gelatin and Asini Corii Colla in Biejiajian pills.Relevant units should pay attention to the content of marker polypeptide of trionycis-shell glue and Asini Corii Colla in Biejiajian pills is lower than the proposed limit.