超高效液相色谱-质谱联用技术检测阿胶中马皮源成分

Petection of equine-hide gelatin in Ejiao by UPLC-MS

目的:建立阿胶中马皮源成分的检测方法,为阿胶药材质量控制提供技术支持。方法:利用高分辨质谱分析方法获得马皮中专属特征肽段,采用胰蛋白酶对阿胶进行酶解,经超高效液相色谱-三重四极杆质谱对马源寡肽段进行检测。电喷雾离子化(ESI),正离子模式下,进行多反应监测,选择马皮特征分子离子峰m/z 386.4(双电荷)→377.3,322.3作为检测离子对。结果:方法学试验考察表明,所建立方法专属性、稳定性、重复性及耐用性均良好,对照物质马源寡肽A线性范围为0.010 86～1.086μg·mL^(-1),相关系数为0.999 9。以所建立的方法对专项抽验的56批阿胶进行检验,结果 28批样品检出马皮源成分。结论:所建立的方法准确可靠,专属性强,可用于阿胶中的马皮源成分的检测。

Objective:To establish an analysis method for specific identification of equine-hide gelatin in Ejiao.Method:The UPLC-MS was used to find the marker peptides for identification of equine-hide gelatin in Ejiao.The sample was analyzed on UPLC coupled with triple quadrupole mass spectrometry after digesting by trypsin.Choosing the characteristic molecular peaks of m/z 386.4 → 377.3,322.3 as target ion pair,collagen marker peptides were detected by mass spectrometry with positive electrospray ionization and multiple reaction monitoring(MRM).Results:The methodological evaluation showed that this method had good specificity,stability,repeatability and durability.The linear ranges of peptide A equine fell within the ranges of 0.010 86-1.086μg·mL^-1 with the correlation coefficient of 0.999 9.The characteristic molecular peaks of equine-hide gelatin were detected in 28 batches of Ejiao.Conclusion:The present method is specific,precise,reliable,and suitable for identification of equine-hide gelatin in Ejiao.