超高效液相色谱-三重四极杆质谱法用于含阿胶中成药中胶类皮源检测研究  

Identification of Gelatin in Traditional Chinese Patent Medicines by UPLC-QqQ-MS

作　　者：李婷[1] 程显隆[2] 王郡瑶 李明华[2] 郭晓晗 李向日[1] 马潇 魏锋[1] 马双成[2] LI Ting;CHENG Xian-long;WANG Jun-yao;LI Ming-hua;GUO Xiao-han;LI Xiang-ri;MA Xiao;WEI Feng;MA Shuang-cheng(Centre of TCM Processing Research,Beijing University of Chinese Medicine,Beijing Key Laboratory for Quality Evaluation of Chinese Materia Medica,Beijing 102488,China;National Institutes for Food and Drug Control,Beijing 100050,China;Gansu Provincial Institute of Drug Control,Lanzhou 730000,China)

机构地区：[1]北京中医药大学中药炮制研究中心,北京市中药品质评价重点实验室,北京102488 [2]中国食品药品检定研究院,北京100050 [3]甘肃省药品检验研究院,兰州730000

出　　处：《中国药学杂志》2023年第3期260-265,共6页Chinese Pharmaceutical Journal

基　　金：国家药品监督管理局中药材及饮片质量控制重点实验室项目资助(2020GSMPA-KL15)。

摘　　要：目的建立含阿胶中成药阿胶补血口服液、阿归养血颗粒中阿胶类皮源检测方法,包括阿胶的专属性检测方法,以及猪皮源、马皮源、牛皮源掺伪检查研究。方法采用胰蛋白酶对用0.01 g·mL^(-1)NH4HCO3溶解的阿归养血颗粒及阿胶补血口服液中阿胶进行酶解,利用超高效液相色谱-三重四极杆质谱(UPLC-QqQ-MS)对样品进行检测。色谱柱为Waters ACQUITY UPLC HSS T3-C_(18)(2.1 mm×100 mm,1.8μm),流动相为体积分数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 mL·min^(-1),柱温30℃,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和m/z 539.8(双电荷)→923.8;马源寡肽A特征分子离子峰m/z 386.4(双电荷)→322.3和m/z 386.4(双电荷)→377.3;新阿胶特征分子离子峰m/z 774.5(双电荷)→799.8和m/z 774.5(双电荷)→1034.6;黄明胶特征分子离子峰m/z 641.3(双电荷)→726.2和m/z 641.3(双电荷)→783.3作为检测离子对。结果10批市售阿归养血颗粒和10批阿胶补血口服液样品中均可检出阿胶的特征肽段,其中的3批阿归养血颗粒样品和2批阿胶补血口服液样品中同时检出了马皮特征肽。结论所建立的方法经方法学验证,专属性强,可用于中成药中阿胶的检测及掺伪检查。OBJECTIVE To establish an analysis method for detection of animal-based gelatin in Chinese patent medicines like Ejiaobuxue oral liquid and Eguiyangxue granules,including the specific identification of dongkey-hide gelatin contained in Chinese patent medicines and the examination of pig-hide gelatin,horse-hide gelatin and bovine-hide gelatin aduteration.METHODS Samples were dissolved in water containing 0.01 g·mL^(-1)NH4HCO3and digested by trypsin.Ultra-high performance liquid chromatography(UPLC)coupled with triple quadruple mass spectrometry(QqQ-MS)was used to detect the samples.Determination was carried out with the application of Waters ACQUITY UPLC HSS T3-C_(18)(2.1 mm×100 mm,1.8μm)column at 30℃.The mobile phase was composed of water containing 0.1%formic acid(A)and acetonitrile(B).Gradient elution(0-25 min,95%A→80%A;25-40 min,80%A→50%A)was conducted at a flow rate of 0.3 mL·min^(-1).Electrospray ionization(ESI)was performed in multiple reaction monitoring(MRM)mode with the transitions of m/z 539.8(double charge)→612.4,m/z 539.8(double charge)→923.8,m/z 386.4(double charge)→322.3 and m/z 386.4(double charge)→377.3,m/z 774.5(double charge)→799.8 and m/z 774.5(double charge)→1034.6,m/z 641.3(double charge)→726.2 and m/z 641.3(double charge)→783.3.RESULTS The characteristic peptides of donkey-hide gelatin could be detected in 20 batches of samples collected from market.Horse skin marker peptide was detected in 3 batches of Eguiyangxue granules and 2 batches of Ejiaobuxue oral liquid.CONCLUSION The established method is specific,which is suitable for identification of donkey-hide gelatin in Chinese patent medicine.

关 键 词：阿归养血颗粒 阿胶补血口服液 特征肽段 专属性鉴别 超高效液相色谱-三重四极杆质谱 

分 类 号：R284[医药卫生—中药学]