超高效液相色谱-四极杆飞行时间质谱快速筛查与确证生鲜牛乳中80种抗生素和激素
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摘要
采用超高效液相色谱-四极杆飞行时间高分辨质谱(UHPLC-QTOF MS)技术建立了生鲜牛乳中80种抗生素和激素的快速筛查方法。牛乳样品经含1%乙酸的乙腈溶液提取,采用QuEChERS(Quick,Easy,Cheap,Effective,Rugged and Safe)方法净化。目标药物经Agilent ZORBAX SB C_(18)色谱柱(3. 0 mm×100 mm,1. 8μm)分离,以乙腈-0. 1%甲酸水溶液为流动相进行梯度洗脱,使用Dual AJS ESI源,在正离子模式下进行数据采集,基质匹配标准曲线法定量。结果表明,80种抗生素和激素类药物在各自的浓度范围内线性关系良好(r~2≥0. 990 0),其定量下限(LOQ,S/N=10)为5～100μg/kg。LOQ加标水平下,80种兽药中23种兽药的回收率为50. 2%～59. 7%,相对标准偏差为1. 1%～13. 0%;其余57种兽药的回收率为61. 7%～119%,相对标准偏差为0. 3%～19. 0%。结合精确分子质量数、保留时间、同位素丰度和二级特征碎片离子对目标化合物进行快速筛查与确证。该方法快速简便、准确、灵敏,适用于牛乳中抗生素和激素类药物残留的高通量筛查与定性鉴定。
A comprehensive analytical method was established for the rapid screening of 80 kinds of antibiotics and hormones in raw milk by ultra performance liquid chromatography-quadrupole timeof-flight mass spectrometry( UHPLC-QTOF MS). The milk samples were extracted with acetonitrile containing 1% acetic acid. The extracts were cleaned up through QuEChERS( Quick, Easy,Cheap,Effective,Rugged and Safe) method. 80 target compounds were separated on an Agilent ZORBAX SB C_(18) column(3. 0 mm × 100 mm,1. 8 μm) by gradient elution using acetonitrile-0. 1% formic acid as mobile phases,and detected by MS under Dual AJS ESI+mode. The analytes were quantified by the matrix-matched calibration standards. As a result,the calibration curves of 80 target compounds were linear in the certain concentration with their correlation coefficients not less than 0. 990 0. The limits of quantitation( LOQ,S/N = 10) for the compounds in milk were in the range of 5-100 μg/kg. The average recoveries for 23 drugs in milk at LOQ spiked levels ranged from 50. 2% to 59. 7% with their relative standard deviations( RSDs) of 1. 1%-13. 0%,while those for the other 57 drugs ranged from 61. 7% to 119% with their RSDs of 0. 3%-19. 0%. The screening and confirmation on the analytes were performed by using precision mass,retention time,isotopic abundance and two-stage ion mass spectra. The method could be applied in the high-throughput screening and identification of antibiotics and hormones in raw milk due to its characteristics of rapidness,simplicity,accuracy and sensitivity.
A comprehensive analytical method was established for the rapid screening of 80 kinds of antibiotics and hormones in raw milk by ultra performance liquid chromatography-quadrupole timeof-flight mass spectrometry( UHPLC-QTOF MS). The milk samples were extracted with acetonitrile containing 1% acetic acid. The extracts were cleaned up through QuEChERS( Quick, Easy,Cheap,Effective,Rugged and Safe) method. 80 target compounds were separated on an Agilent ZORBAX SB C_(18) column(3. 0 mm × 100 mm,1. 8 μm) by gradient elution using acetonitrile-0. 1% formic acid as mobile phases,and detected by MS under Dual AJS ESI+mode. The analytes were quantified by the matrix-matched calibration standards. As a result,the calibration curves of 80 target compounds were linear in the certain concentration with their correlation coefficients not less than 0. 990 0. The limits of quantitation( LOQ,S/N = 10) for the compounds in milk were in the range of 5-100 μg/kg. The average recoveries for 23 drugs in milk at LOQ spiked levels ranged from 50. 2% to 59. 7% with their relative standard deviations( RSDs) of 1. 1%-13. 0%,while those for the other 57 drugs ranged from 61. 7% to 119% with their RSDs of 0. 3%-19. 0%. The screening and confirmation on the analytes were performed by using precision mass,retention time,isotopic abundance and two-stage ion mass spectra. The method could be applied in the high-throughput screening and identification of antibiotics and hormones in raw milk due to its characteristics of rapidness,simplicity,accuracy and sensitivity.
引文
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[26]GB/T 20759-2006.Method for the Determination of Sixteen Sulfonamide Residues in Livestock and Poultry Muscles-LC-MS/MS Method.National Standard of the Peoples Republic of China(畜禽肉中十六种磺胺类药物残留量的测定液相色谱-串联质谱法.中华人民共和国国家标准).
[27]GBT 20366-2006.Method for the Determination of Quinolones in Animal Tissues-LC-MS/MS Method.National Standard of the Peoples Republic of China(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.中华人民共和国国家标准).
[28]GBT 21311-2007.Determination of Residues of Nitrofuran Metabolites in Foodstuffs of Animal Origin-HPLC-MS/MSMethod.National Standard of the Peoples Republic of China(动物源性食品中硝基呋喃类药物代谢物残留量检测方法高效液相色谱-串联质谱法.中华人民共和国国家标准).
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[3]Li S J,Qiu Y M,Wang C.Veterinary Drug Residue Analysis.Shanghai:Shanghai Science and Technology Press(李俊锁,邱月明,王超.兽药残留分析.上海:上海科学技术出版社),2002.
[4]Andrew S M,Moyes K M,Borm A A,Fox L K,Leslie K E,Hogan J S,Oliver S P,Schukken Y H,Owens W E,Norman C.J.Veter.Microbiol.,2009,134(1/2):150-156.
[5]Sun S L.Environmental Hormones and the Future of Mankind.Beijing:Chemical Industry Press(孙胜龙.环境激素与人类未来.北京:化学工业出版社),2005.
[6]Zhu W X,Liu Y F,Yuan P,Yang J Z.Chin.J.Chromatogr.(祝伟霞,刘亚风,袁萍,杨冀州.色谱),2010,28(11):1031-1037.
[7]Wang J,Wang Q L,Ye J M,Chen Q J.Chin.J.Anal.Lab.(王京,王庆龄,叶佳明,陈青俊.分析试验室),2016,35(8):955-960.
[8]Li S Y,Zhang H Q,Zheng C Y,Zhou F,Lin J,Wang Y.J.Chin.Fishery Qual.Stand.(李诗言,张海琪,郑重莺,周凡,林静,王扬.中国渔业质量与标准),2015,5(1):42-51.
[9]Li R,Yang L Q,Zhang P Y,Luo Y D,Zhang P J,Gao Y Q.Chin.J.Chromatogr.(李蓉,杨璐齐,张鹏云,罗阳丹,张朋杰,高永清.色谱),2017,35(12):1266-1275.
[10]Wu J P,Zhang X,Li D N,Yan F,Zhou Y R,Gu X.J.Instrum.Anal.(吴剑平,张鑫,李丹妮,严凤,周悦榕,顾欣.分析测试学报),2015,34(5):610-615.
[11]Wang L,Li Y Q,Wang H B,Guan Y L.Chin.J.Anal.Chem.(王炼,黎源倩,王海波,官艳丽.分析化学),2011,39(2):203-207.
[12]Qu Y,Lu Y,Feng N,Zhao J P,Jin W W,Wang G S.Food Sci.(渠岩,路勇,冯楠,赵俊平,金伟伟,王贵双.食品科学),2012,33(8):252-255.
[13]Cao Y F,Kang J,Chang Q Y,Hu X Y,Wang Z B,Fan C L,Wang M L.Chin.J.Chromatogr.(曹亚飞,康健,常巧英,胡雪艳,王志斌,范春林,王明林.色谱),2015,33(2):132-139.
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[19]Lin X L,Li N,Huo F,Dong Y F,Peng W.J.Instrum.Anal.(林小莉,李宁,霍峰,董艳峰,彭玮.分析测试学报),2016,35(3):322-326.
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[21]Patyra E,Kwiatek K.J.Vet.Res.,2016,60:35-41.
[22]Jadhav M,Thekkumpurath A S,Nakade M,Gadgil M,Oulkar D,Arimboor R,Ramakrishna M,Banerjee K.J.AOACInt.,2017,100(3):603-609.
[23]Dasenaki M E,Michali C S,Thomaidis N S.J.Chromatogr.A,2016,1452:67-80.
[24]Li S,Chen S B,Li L Q,Xu X W,Chen X F,Zhong Y Y,Zhang Y H,Wang C X.Chin.J.Anal.Chem.(李双,陈树兵,李露青,徐旭文,陈先锋,钟莺莺,张雅珩,王传现.分析化学),2018,36(6):506-517.
[25]Xie H B,Wang Y G,Zhou M Y,Zhao H F,Jiang W F,Deng S,Dong S Z.J.Instrum.Anal.(谢寒冰,王毅刚,周明莹,赵海峰,蒋万枫,邓书,董诗竹.分析测试学报),2014,33(4):373-379.
[26]GB/T 20759-2006.Method for the Determination of Sixteen Sulfonamide Residues in Livestock and Poultry Muscles-LC-MS/MS Method.National Standard of the Peoples Republic of China(畜禽肉中十六种磺胺类药物残留量的测定液相色谱-串联质谱法.中华人民共和国国家标准).
[27]GBT 20366-2006.Method for the Determination of Quinolones in Animal Tissues-LC-MS/MS Method.National Standard of the Peoples Republic of China(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.中华人民共和国国家标准).
[28]GBT 21311-2007.Determination of Residues of Nitrofuran Metabolites in Foodstuffs of Animal Origin-HPLC-MS/MSMethod.National Standard of the Peoples Republic of China(动物源性食品中硝基呋喃类药物代谢物残留量检测方法高效液相色谱-串联质谱法.中华人民共和国国家标准).
[29]Ortelli D,Cognard E,Jan P,Edder P.J.Chromatogr.B,2009,877:2363-2374.
[30]Gao F D,Zhao Y,Shao B,Zhang J.Chin.J.Chromatogr.(高馥蝶,赵妍,邵兵,张晶.色谱),2012,30(6):560-567.
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