基于分子印迹技术检测鸡肉中16种喹诺酮药物残留
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摘要
建立了同时测定鸡肉中氧氟沙星、诺氟沙星、培氟沙星、环丙沙星、洛美沙星、达氟沙星、恩诺沙星、沙拉沙星、二氟沙星、吡哌酸、麻保沙星、依诺沙星、氟罗沙星、奥比沙星、尤利沙星、加替沙星16种喹诺酮类药物残留的分子印迹固相萃取(MISPE)-超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法。样品均质处理后,用磷酸盐缓冲溶液提取,经MISPE柱上样,依次用水、乙腈、2%乙酸乙腈、0.1%氨水淋洗,5%氨化甲醇洗脱,用BEHC18柱分离,以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,电喷雾正离子多反应监测模式下,外标法定量。实验对上样液、淋洗与洗脱液的条件进行优化,结果表明,16种喹诺酮类药物检出限为0.08μg/kg,定量限为0.25μg/kg;药物浓度在0.25～200μg/L范围内相关系数(r)均大于0.99;在0.25、2.5、5.0μg/kg三个添加水平下,各药物回收率为88.6%～101.9%,相对标准偏差(RSD)为2.9%～10.5%。该方法灵敏度高,操作简单、快速。
A ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)-molecularly imprinted solid-phase extraction(MISPE) method was established for the simultaneous determination of 16 quinolone drug residues(ofloxacin,norfloxacin,pefloxacin,ciprofloxacin,lomefloxacin,danofloxacin,enrofluxacin,sarafloxacin,difluoxacin pipemidic acid,marbofloxacin,enoxacin,fleroxacin,orbifloxacin,ulifloxacin,gatifloxacin).The conditions of the MISPE column of upper sample solution,leaching and eluting were optimized.The samples were extracted with phosphate buffer solution,added on the MISPE column.The column was washed by water,acetonitrile,2%acetic acid-acetonitrile and 0.1%ammonia leaching,5%ammoniated methanol elution.The analytes were separated by Acquity LC BEH C18 column with acetonitrile-0.1%formic acid for gradient elution,detected by electrospray positive ion multi-reaction monitoring(MRM) mode.The limits of detection were 0.08 μg/kg and the limits of quantification were 0.25 μg/kg.The linear ranges were 0.25-200 μg/L,with correlation coefficients(r) greater than 0.99.At the spiked levels of 0.25,2.5 and 5.0 μg/kg,the recoveries were 88.6%-101.9%,and RSDs were 2.9%-10.5%.The method is sensitive,simple and fast for the determination of 16 quinolone drug residues.
A ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)-molecularly imprinted solid-phase extraction(MISPE) method was established for the simultaneous determination of 16 quinolone drug residues(ofloxacin,norfloxacin,pefloxacin,ciprofloxacin,lomefloxacin,danofloxacin,enrofluxacin,sarafloxacin,difluoxacin pipemidic acid,marbofloxacin,enoxacin,fleroxacin,orbifloxacin,ulifloxacin,gatifloxacin).The conditions of the MISPE column of upper sample solution,leaching and eluting were optimized.The samples were extracted with phosphate buffer solution,added on the MISPE column.The column was washed by water,acetonitrile,2%acetic acid-acetonitrile and 0.1%ammonia leaching,5%ammoniated methanol elution.The analytes were separated by Acquity LC BEH C18 column with acetonitrile-0.1%formic acid for gradient elution,detected by electrospray positive ion multi-reaction monitoring(MRM) mode.The limits of detection were 0.08 μg/kg and the limits of quantification were 0.25 μg/kg.The linear ranges were 0.25-200 μg/L,with correlation coefficients(r) greater than 0.99.At the spiked levels of 0.25,2.5 and 5.0 μg/kg,the recoveries were 88.6%-101.9%,and RSDs were 2.9%-10.5%.The method is sensitive,simple and fast for the determination of 16 quinolone drug residues.
引文
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[18] GB/T 20366-2066.Determination of Quinolone Residues in Animal Products by Liquid Chromatography-Tandem Mass Spectrometry(动物源产品中喹诺酮类残留量的测定液相色谱-串联质谱法).
[2] Tyson G H,Tate H P,Zhao S,et al.Antimicrobial Agents & Chemotherapy,2017,61(10):13.
[3] LI W W.Chinese Journal of Convalescent Medicine(黎微微.中国疗养医学),2013,22(12):1090.
[4] (EU) No 37/2010 On Pharmacologically Active Substances and Their Classification Regarding Maximum Residue Limits in Foodstuffs of Animal Origin.
[5] 21CFR556 Tolerances for Residues of New Animal Drugs in Food.
[6] Ministry of Agriculture.No.235 Bulletin of the Ministry of Agriculture of the People’s Republic of China(农业部.中华人民共和国农业部公告第235号).
[7] LIANG Y,XIONG M J,ZHANG J J,et al.West China Medical Journal(梁艳,熊梅瑾,张静静,等.华西药学杂志),2012,27(6):715.
[8] Zhang J,Liu D,Shi Y,et al.Journal of Chromatography B,2017,1068:24.
[9] ZHANG Y,ZHOU W E,LI S H,et al.Science and Technology of Food Industry(张元,周伟娥,李绍辉,等.食品工业科技),2016,37(5):378.
[10] Zhang X,Wang C,Yang L,et al.Journal of Chromatography B Analytical Technologies in the Biomedical & Life Sciences,2017,1064:68.
[11] Liu S T,Yan H Y,Wang M Y.Journal of Agricultural and Food Chemistry,2013,61:97.
[12] Zheng Y Q,Liu Y H,Guo H B,et al.Journal of Chromatography A,2011,690(2):269.
[13] WANG L J,CAO X M,SUN X L,et al.Chinese Journal of Chromatography(王丽君,曹旭敏,孙晓亮,等.色谱),2016,34(3):240.
[14] Tang Y W,Li M,Gao X,et al.Microchimica Acta,2016,183(2):589.
[15] Urraca J L,Castellari M,Barrios C A,et al.Journal of Chromatography A,2014,1343:1.
[16] WANG L Q,HE L M,ZENG Z L,et al.Journal of Mass Spectrometry(王立琦,贺利民,曾振灵,等.质谱学报),2011,32(6):321.
[17] LIN F,LIN H D,WU Y X,et al.Journal of Instrumental Analysis(林峰,林海丹,吴映璇,等.分析测试学报),2004,23(5):43.
[18] GB/T 20366-2066.Determination of Quinolone Residues in Animal Products by Liquid Chromatography-Tandem Mass Spectrometry(动物源产品中喹诺酮类残留量的测定液相色谱-串联质谱法).