超高效液相色谱-三重四极杆质谱法检测复方阿胶浆中阿胶

目的: 建立复方阿胶浆中阿胶的专属性检测方法。方法: 采用胰蛋白酶对复方阿胶浆中阿胶成分进行酶解,利用超高效液相色谱-三重四极杆质谱(RRLC-QQQ/MS)对阿胶的专属性特征分子离子峰进行检测。采用Agilent SB-C₁₈ (2.1 mm×100 mm,1.8 μm)色谱柱,以0.1%甲酸水溶液为流动相A,以乙腈为流动相B,梯度洗脱(0~25 min,95%A→80%A,5%B→20%B;25~40 min,80%A→50%A,20%B→50%B)。流速0.3 mL · min^-1,柱温40 ℃,进样量5 μL;选择阿胶特征分子离子峰m/z 539.8(双电荷)→612.4和m/z 539.8(双电荷)→923.8作为检测离子对,离子化模式为ESI⁺,进行多反应监测。结果: 3批市售样品中均可检出阿胶的特征分子离子峰,即同时检出m/z 539.8(双电荷)→612.4和m/z 539.8(双电荷)→923.8离子对。结论: 所建立的方法经方法学验证,阴性样品及其他胶类如黄明胶、龟甲胶和鹿角胶等对复方阿胶浆样品测定无干扰,样品检出浓度为含5%阿胶的复方阿胶浆样品。方法专属性强,可用于复方阿胶浆中阿胶的检测。

Objective: To establish an analytical method for identification of donkey-hide gelatin in Fufang Ejiao Jiang. Methods: Identification of donkey-hide gelatin in Fufang Ejiao Jiang was established by rapid resolution liquid chromatography(RRLC)coupled to triple quadruple mass spectrometry(QQQ-MS).Chromatographic separation was carried out on an Agilent Zorbax SB-C₁₈ reversed phase analytical column(100 mm×2.1 mm, 1.8 μm particle size)at a column temperature of 40℃.The injected sample volume was 5 μL.The mobile phase consisted of 0.1% formic acid in water(eluent A)and acetonitrile(eluent B).The gradient elution was performed as follows:0- 25 min eluent A 95%→80%, elutent B 5%→20%;25-40 min, eluent B 20%→50%.A flow rate of 0.3 mL · min^-1 was employed for elution.Collagen marker peptides were detected by mass spectrometry in the positive electrospray ionization with multiple reaction monitoring(MRM). Results: The characteristic molecular peaks of donkey-hide gelatin m/z 539.8→612.4 and m/z 539.8→923.8 were detected in all three batches of commercial samples. Conclusion: The methodological verification showed that the donkey-hide gelatin was not interfered with the negative gelatins and other gelatins such as bovine-hide gelatin, deer-horn glue and tortoise shell gelatin.The detection limit of the new method extended to 5%.The present method was specific, precise and reliable, and was suitable for identification of donkey- hide gelatin in Fufang Ejiao Jiang.