超声辅助酶解与串联质谱联用检测阿胶中马皮成分  

Detection of Horse-Hide Gelatin in Asini Corii Colla by Using Ultrasonic Assisted Enzyme Digestion Combined with Tandem Mass Spectrometry Method

作　　者：蔡朔 蒋梦彤 赵珂璇 韩疏影[1] 费菲 曹蓓 赵明 段金廒[1,3] 刘睿[1,3,4] CAI Shuo;JIANG Meng-tong;ZHAO Ke-xuan;HAN Shu-ying;FEI Fei;CAO Bei;ZHAO Ming;DUAN Jin-ao;LIU Rui(School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, China;Phase Ⅰ Clinical Trials Unit, Nanjing Drum Tower Hospital, Nanjing, 210008, China;Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing University of Chinese Medicine, Nanjing, 210023, China;Jiangsu Key Laboratory of Research and Development in Marine Bio-Resource Pharmaceutics, Nanjing, 210023, China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]南京鼓楼医院Ⅰ期临床试验室,江苏南京210008 [3]南京中医药大学江苏省中药资源产业化过程协同创新中心/中药资源产业化与方剂创新药物国家地方联合工程研究中心,江苏南京210023 [4]江苏省海洋药用生物资源研究与开发重点实验室,江苏南京210023

出　　处：《南京中医药大学学报》2020年第5期623-628,共6页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81973450);国家重点研发计划(2018YFC1706100);江苏省高校“青蓝工程”优秀青年骨干教师(2016年);江苏省“333工程”第三层次(2016年);中华中医药学会“青年人才托举工程”。

摘　　要：目的超声辅助快速酶解与串联质谱联用实现准确、高效检测阿胶中的马皮成分,为阿胶质量标准研究提供实验依据。方法采用超声辅助胰蛋白酶酶解的方法,快速制备阿胶酶解液样品,采用超高效液相色谱-三重四极杆串联质谱(UPLC-QQQ-MS)对阿胶酶解液进行检测,在电喷雾离子化的正离子模式下,多反应监测模式(MRM),以驴专属肽m/z 691.0→809.4、马专属肽m/z 698.3→809.4为离子对,检测阿胶样品中的马专属肽,以及驴专属肽与马专属肽的相对峰面积比例。结果采用的方法可在20 min内准确判别阿胶中是否掺有马皮成分,及掺入的马皮成分的比例,马皮成分的检出限为1%。结论超声辅助快速酶解与串联质谱联用的方法准确可靠、专属性强、检测效率高,可用于阿胶中马皮成分的快速检测。OBJECTIVE To accurately and efficiently detect the horse hide components in Asini Corii Colla(ACC)by using ultrasonic assisted fast enzyme digestion and tandem mass spectrometry,and provide experimental basis for ACC quality standard investigation.METHODS By using ultrasonic assisted trypsin digestion method,ACC hydrolysate samples were prepared rapidly.High performance liquid chromatography with three quadrupole mass spectrometry was used to detect the ACC hydrolysate samples.Under positive electrospray ionization mode and multiple reaction monitoring(MRM),donkey-specific peptide(Pep-1)with ion pair of m/z 691.0→809.4 and horse-specific peptide(Pep-2)with ion pair of m/z 698.3→809.4 were used to detect Pep-2 in ACC,and the relative peak area ratio between Pep-2 and Pep-1.RESULTS This method could be used to determine if there was horse-hide gelatin(HHG)in ACC within 20 min,and the proportion of HHG in ACC.The detection limit of HHG adulteration was low to 1%.CONCLUSION Ultrasound-assisted fast enzyme digestion combined with tandem mass spectrometry showed good accuracy,reliability,specificity and efficiency,and can be used for the rapid detection of HHG in ACC.

关 键 词：阿胶 马皮成分 超声辅助酶解 串联质谱 专属肽 

分 类 号：R284[医药卫生—中药学]