基于超高效液相色谱串联质谱法特征肽检测技术的阿胶鉴别与6种动物异源性成分检查  

Identification of Colla Corii Asini and Examination of Six Animal Heterologous Components Based on UPLC-MS/MS Characteristic Peptide Detection Technology

作　　者：潘志文 黄婉锋 钱鑫 周宗洲 PAN Zhiwen;HUANG Wanfeng;QIAN Xin;ZHOU Zongzhou(Foshan Food and Drug Inspection and Testing Center,Foshan,Guangdong,China 528051)

机构地区：[1]广东省佛山市食品药品检验检测中心,广东佛山528051

出　　处：《中国药业》2024年第11期79-83,共5页China Pharmaceuticals

摘　　要：目的 建立同时检测阿胶中马、牛、猪、骆驼、羊、鹿6种动物皮异源性成分的超高效液相色谱串联质谱(UPLC-MS/MS)法。方法 阿胶粉末加1%碳酸氢铵溶液超声溶解,滤过,采用胰蛋白酶酶解15 h。采用UPLC-MS/MS法,以阿胶特征肽段质荷比(m/z)539.8→924.3和驴源特征肽m/z 570.5→698.1、马源寡肽A m/z 386.4→377.3、牛皮特征肽A m/z 641.2→726.3、猪皮特征肽m/z 774.5→1 035.7、骆驼皮特征肽m/z 603.8→281.8、羊皮特征肽m/z 553.0→450.8、鹿皮特征肽m/z 733.1→962.5作为检测离子对,电喷雾电离(ESI)离子源、正离子模式进行多反应监测(MRM),对10批药用阿胶、6批食品级阿胶样品进行质量评价。结果 所选各动物源特征肽段均只在对应动物皮胶中检出,方法专属性强;方法灵敏度符合质量控制要求;酶解重复性、进样精密度及样品稳定性均满足方法学验证要求,结果的RSD均小于9.0%(n=6)。6批食用级阿胶样品中均未检出阿胶成分,判定主要成分为牛皮源、骆驼皮源、马皮源、猪皮源成分。8批药用阿胶检出阿胶成分,其中1批阿胶成分含量偏低,1批检出马皮源成分;2批药用阿胶未检出阿胶成分,判定为假冒产品。结论 所建立的6种动物皮胶检测方法的专属性强、灵敏度高,可用于阿胶鉴别及掺假杂皮胶的检测。Objective To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method to detect the six animal heterologous components such as horse hide glue,cattle hide glue,pig hide glue,camel hide glue,sheep hide glue,deer hide glue in Colla Corii Asini.Methods Colla Corii Asini powder was dissolved with 1%ammonium bicarbonate solution for ultrasonic dissolution,filtration,and enzymolysis with trypsin for 15 h.UPLC-MS/MS method was adopted,with the Colla Corii Asini characteristic peptide[mass charge ratio(m/z)539.8→924.3],donkey characteristic peptide(m/z 570.5→698.1),horse characteristic peptide A(m/z 386.4→377.3),cattle characteristic peptide A(m/z 641.2→726.3),pig characteristic peptide(m/z 774.5→1035.7),camel characteristic peptide(m/z 603.8→281.8),sheep characteristic peptide(m/z 553.0→450.8),and deer characteristic peptide(m/z 733.1→962.5)as detection ion pairs.Multi-reaction monitoring(MRM)was adopted with apositive ion mode of electric spray ion source(ESI).The quality of 10 batches of medicinal Colla Corii Asini samples and six batches of food-grade Colla Corii Asini samples were evaluated.Results The selected characteristic peptide segments of each animal were only detected in the corresponding animal hide glue,and the specificity of the method was strong.The sensitivity of the method met the quality control requirements.The repeatability,injection precision,and sample stability of enzymolysis all met the requirements of methodological validation,and the RSDs of the results were all lower than 9.0%(n=6).In six batches of food-grade Colla Corii Asini samples,no Colla Corii Asini component was detected and the main components were determined to be cattle hide glue,camel hide glue,horse hide glue and pig hide glue.The Colla Corii Asini was detected in eight batches of medicinal Colla Corii Asini samples,of which the content of Colla Corii Asini in one batch was low,and horse hide glue was detected in one batch.Two batches of medicinal Colla Corii Asini were judge

关 键 词：超高效液相色谱串联质谱法 阿胶 特征肽 动物皮胶 异源性成分 

分 类 号：R932[医药卫生—生药学] R284.1[医药卫生—药学] R286.0