基于特征多肽的地龙鉴别与伪品保宁腔蚓检查方法的建立及应用  

Development and application of a method for identifying Pheretima and a common counterfeit of Metaphire magna based on signature peptides

作　　者：刘蕊[1,2] 张静娴 胡青 孙健 于泓 冉莹瑛 黄帆 毛秀红 季申 LIU Rui;ZHANG Jing-xian;HU Qing;SUN Jian;YU Hong;RAN Ying-ying;HUANG Fan;MAO Xiu-hong;JI Shen(Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Shanghai Institute for Food and Drug Control,Shanghai 201203,China)

机构地区：[1]上海中医药大学,上海201203 [2]上海市食品药品检验研究院,上海201203

出　　处：《药学学报》2024年第10期2842-2848,共7页Acta Pharmaceutica Sinica

基　　金：上海市中药和保健食品品质与安全检测专业技术服务平台(21DZ2290200);2024年国家药品标准提高项目地龙(2024Z010);上海市人才发展资金(2021085)。

摘　　要：基于种属特异性多肽,采用超高效液相色谱-三重四极杆质谱(UHPLC-MS/MS)建立了地龙鉴别和伪品保宁腔蚓的检查方法,并应用于地龙及两种成方制剂的检测分析。采用CORTECS T3 C18色谱柱分离,以0.1%甲酸溶液-乙腈为流动相,梯度洗脱,采用ESI+离子源、多重反应监测(MRM)模式,同时监测3组离子对。结果表明,本方法具有良好的专属性,可以鉴定出广地龙、沪地龙、保宁腔蚓,鉴定结果与DNA条形码分子鉴定结果一致。掺伪检查研究表明,保宁腔蚓多肽M的检出限为1μg·g^(-1),广地龙中掺入1%保宁腔蚓时即可检出,方法灵敏度高。54批市售地龙样品中广地龙占比35%、沪地龙35%、保宁腔蚓15%,另外15%未检出任何目标离子,经DNA条形码分子鉴定发现这些样品主要来源为远盲蚓属,部分样品性状与沪地龙相似。成方制剂分析结果显示,3批小活络丸(3/6)未检出任何多肽离子,2批伸筋丹胶囊(2/4)中检出保宁腔蚓。本研究所建立的方法专属性好、灵敏度高、可行性强可以用于地龙及其相关制剂的质量控制研究中,对于提高质量标准、规范地龙药材市场具有重要意义。Based on the species-specific peptides of Pheretima and its common counterfeit(Metaphire magna),an identification method was established using ultra-high performance liquid chromatography tandem triple quadrupole mass spectrometry(UHPLC-MS/MS)for quality evaluation of Pheretima and its preparations.Separation was performed on a CORTECS T3 C18 column with 0.1%formic acid and acetonitrile as the mobile phases.Mass spectrometry with multiple reaction monitoring(MRM)using ESI+mode was used to simultaneously monitor three ion pairs.The results indicated that the method was specific and could distinguish Guang Dilong,Hu Dilong,and M.magna,which were consistent with those of DNA barcode identification.The adulteration test showed that the LOD of peptide M was 1μg·g^(-1).Peptide M could be detected when 1%M.magna was added to Guang Dilong,indicating the high sensitivity of the method.Fifty-four batches of commercially available samples contained 35%Guang Dilong,35%Hu Dilong,and 15%M.magna.No ions were detected in 15%of the samples,and DNA barcode identification revealed that they were mainly from Amynthas,with similar appearance to Hu Dilong.The analysis results of the formulation showed that no peptide ions were detected in 3 batches of Xiaohuoluo pills(3/6)and M.magna were detected in 2 batches of Shenjindan capsules(2/4).The developed method in the study has good specificity,sensitivity,and feasibility,and could be used for quality control of Pheretima and its related preparations.It is of great significance for improving quality standards and regulating the medicinal market of Pheretima.

关 键 词：地龙 保宁腔蚓 特征多肽 质量标准 小活络丸 伸筋丹胶囊 

分 类 号：R917[医药卫生—药物分析学]