超高效液相色谱-三重四极杆质谱法检测孕康颗粒中阿胶的特征肽及掺杂猪皮源成分

Determination of Characteristic Peptide of Donkey-hide Gelatin and Ingredients from Pig Skin in Yunkang Granules by UPLC-QQQ MS

目的:建立孕康颗粒中阿胶的特征肽成分及掺杂猪皮源成分的专属性检测方法。方法:采用胰蛋白酶对孕康颗粒进行酶解,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及猪皮源成分进行检测。色谱柱:Agilent Zorbax Eclipse-C_(18)柱(2.1 mm×100 mm,1.5μm);流动相:乙腈(A)-0.1%甲酸溶液(B),梯度洗脱;流速:0.3 ml·min^(-1);柱温:40℃。选择阿胶特征分子离子峰m/z 539.8(双电荷)→612.4,923.8及猪皮源特征离子峰m/z 774.5(双电荷)→977.8,1034.6作为检测离子对,离子化模式为ESI+,进行多反应监测。结果:8批市售孕康颗粒样品中均可检出阿胶的特征肽段,均未检出猪皮源成分。结论:建立的方法专属性强、准确可靠,可用于孕康颗粒中阿胶特征肽成分及掺杂猪皮源成分的检测。

Objective:To establish an analysis method for the determination of characteristic peptide of donkey-hide gelatin and ingredients from pig skin in Yunkang granules.Methods:Determination of the components from donkey-hide gelatin and ingredients from pig skin in Yunkang granules was performed by ultra performance liquid chromatography(UPLC)coupled with triple quadruple mass spectrometry(QQQ MS).Determination was carried out on an Agilent Zorbax Eclipse-C_(18)(2.1 mm×100 mm,1.5μm)column with the column temperature at 40℃.The mobile phase was composed of acetonitrile(A)and 0.1% formic acid(B)with gradient elution at a flow rate of 0.3 ml·min^(-1).The characteristic ion pairs of donkey-hide gelatin m/z 539.8(double charge)→612.4,923.8were as the detection ion pairs and the characteristic ion pairs of ingredients from pig skin m/z 774.5(double charge)→977.8,1034.6 were as the detection ion pairs.The positive electrospray ionization and multiple reaction monitoring were adopted.Results:Donkey-hide gelatin was detected out in eight batches of samples,and ingredients from pig skin were found at the same time.Conclusion:The method is highly specific,reliable and accurate,which can be applied to detect donkey-hide gelatin and ingredients from pig skin in Yunkang granules.