基于通过式固相萃取净化的液相色谱-串联质谱法测定猪肉中5种肽类抗生素  

Determination of Five Peptide Antibiotics in Pork by HPLC-MS/MS Based on Pass-through Solid Phase Extraction

作　　者：刘真 王玉梅 胡文彦 LIU Zhen;WANG Yu-mei;HU Wen-yan(Nanjing Institute for Food and Drug Control,Nanjing 211198,China)

机构地区：[1]南京市食品药品监督检验院,江苏南京211198

出　　处：《分析测试学报》2023年第2期197-203,共7页Journal of Instrumental Analysis

基　　金：国家市场监督管理总局科技计划项目(2020MK140)。

摘　　要：建立了一种基于免活化通过式固相萃取净化技术的液相色谱-串联质谱法(HPLC-MS/MS)用于测定猪肉中5种肽类抗生素(达托霉素、万古霉素、去甲万古霉素、维吉尼亚霉素M1、维吉尼亚霉素S1)。猪肉采用甲醇-2%甲酸(体积比为5∶2)进行提取,经免活化的Captiva EMR-Lipid固相萃取小柱净化后,以Phenomenex Kinetex F5(2.1 mm×100 mm,2.6μm)色谱柱分离,0.1%甲酸乙腈溶液和0.1%甲酸梯度洗脱。采用电喷雾离子源正离子模式扫描,多反应模式(MRM)监测目标物,内标法定量。结果表明,猪肉中达托霉素在2~200 ng/mL范围内,万古霉素及去甲万古霉素在5~500 ng/mL范围内,维吉尼亚霉素M1及维吉尼亚霉素S1在1~100 ng/mL范围内均呈良好线性关系,相关系数(r^(2))均大于0.99,5种肽类抗生素的检出限为3~7.5μg/kg,定量下限为5~25μg/kg。在25、50、250μg/kg 3个加标水平下的平均回收率为80.2%~102%,日内相对标准偏差(Intra-RSD)为1.3%~7.4%,日间相对标准偏差(Inter-RSD)为2.6%~8.5%。该方法基于Captiva EMR-Lipid固相萃取小柱免活化净化,前处理简便,回收率好,重复性佳,适用于猪肉中5种肽类抗生素的同时检测。A high performance liquid chromatography-tandem mass spectrometric(HPLC-MS/MS) method was developed for the determination of five peptide antibiotics,i. e.daptomycin,vancomycin, norvancomycin, virginiamycin M1 and virginiamycin S1 in pork based on a purification technique with non-activation pass-through solid phase extraction(SPE) column.The pork samples were extracted with methanol-2% formic acid(5∶2,by volume),and purified with a non-activation Captiva EMR-Lipid SPE column.The separation was carried out on a Phenomenex Kinetex F5(2. 1mm × 100 mm,2. 6 μm) column by gradient elution,with 0. 1% formic acid acetonitrile solution and 0. 1% formic acid as mobile phases. The target substances were analyzed in electrospray ion source positive ion mode under multi-reaction monitoring(MRM) mode,and quantified by the internal standard method.Results showed that the calibration curves of five peptide antibiotics in pork were linear(r~2 > 0. 99) in the range of 2-200 ng/mL for daptomycin,5-500 ng/mL for vancomycin and norvancomycin,and 1-100 ng/mL for virginiamycin M1 and virginiamycin S1.The limits of detection were in the range of 3-7. 5 μg/kg,and the limits of quantitation were in the range of 5-25 μg/kg.The recoveries of five peptide antibiotics from negative pork samples at three spiked levels of 25, 50, 250 μg/kg ranged from 80. 2% to 102%,with intra-RSDs and inter-RSDs of 1. 3%-7. 4% and 2. 6%-8. 5%,respectively.With the characteristics of simple pretreatment,good recovery and repeatability,this method based on Captiva EMR-lipid SPE column is suitable for the simultaneous analysis of 5 peptide antibiotics in pork.

关 键 词：肽类抗生素 液相色谱-串联质谱法(HPLC-MS/MS) 猪肉 通过式固相萃取 

分 类 号：O657.63[理学—分析化学] O629.5[理学—化学]