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
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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。Abstract: 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.
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