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

关键词:  鳖甲胶  阿胶  超高效液相色谱-串联质谱法  多肽  含量

Key words:  trionycis-shell glue  Asini Corii Colla  UHPLC-MS/MS  polypeptide  content

摘要:

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。

Abstract:

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.