超高效液相色谱-三重四极杆质谱法检测阿胶补血颗粒中的阿胶

RRLC/QQQ-MS analysis of donkey-hide gelatin in Ejiao Buxue granula

目的:建立阿胶补血颗粒中阿胶的专属性检测方法。方法:采用胰蛋白酶对阿胶补血颗粒中阿胶进行酶解,利用超高效液相色谱-三重四极杆质谱(UPLC-QQQ MS)对阿胶的专属性特征肽段进行检测。色谱柱为Agilent SB-C_(18)(2.1 mm×100 mm,1.8μm),流动相为0.1%甲酸溶液(A)-乙腈(B),梯度洗脱(0～25 min,95%A→80%A;25～40 min,80%A→50%A),流速0.3 m L·min^(-1),柱温40℃,进样量5μL;离子化模式为ESI+,进行多反应监测,选择阿胶特征分子离子峰m/z 539.8(双电荷)→612.4,m/z 539.8(双电荷)→923.8作为检测离子对。结果:3批市售样品中均可检出阿胶的特征肽段。结论:所建立的方法经方法学验证,专属性强,可用于阿胶补血颗粒中阿胶的检测。

Objective：To establish an analysis method for specificity identification of donkey-hide gelatin in Ejiao Buxue granula.Methods：Samples were dissolved by water containing 1% NH_4HCO_3 and digested by trypsin.Ultra-high performance liquid chromatography（UPLC）coupled to triple quadruple mass spectrometry（QQQ MS）was used to detected the specific marker peptides in donkey-hide gelatins.Determination was carried out with the application of an Agilent SB-C_（18）（2.1 mm×100 mm,1.8 μm）column at temperature of 40 ℃.The mobile phase was composed of water solvent containing 0.1% formic acid（A）and acetonitrile（B）with gradient elution（0-25 min,95%A → 80%A;25-40 min,80%A → 50%A）at a flow rate of 0.3 m L·min_（-1）.The electrospray ionization（ESI）source was performed in multiple reaction monitoring（MRM）mode of the transitions of m/z 539.8（double charge）→ 612.4,m/z 539.8（double charge）→ 923.8.Results：The marker peptides can be detected in all samples by this method.Conclusion：The established method is specific,which is suitable for identification of donkey-hide gelatin in Ejiao Buxue granula.