TurboFlow在线净化-液相色谱-串联质谱法测定畜禽肉中20种(氟)喹诺酮类兽药残留量
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摘要
建立TurboFlow在线净化-液相色谱-串联质谱测定畜禽肉中20种(氟)喹诺酮类药物((Fluoro)quinolones,(F)QNs)残留量的分析方法。样品用甲醇-乙腈(1∶1,V/V)进行均质提取,Cyclone-p在线净化柱净化富集提取液中的(F)QNs,然后将富集所得分析物洗脱,转至Hypersil Gold C_8分析柱,经色谱分离后,用串联质谱检测。结果表明:西诺沙星、伊诺沙星和丹诺沙星在0.2～20.0 ng/mL的线性范围内线性良好,其余17种(F)QNs在0.1～10.0 ng/mL的线性范围内线性良好,且R~2均大于0.99;西诺沙星、伊诺沙星和丹诺沙星的定量限为2μg/kg,其余17种(F)QNs为1μg/kg;20种(F)QNs在3个水平的添加回收率为85.4%～108.2%,相对标准偏差为4.04%～8.57%。方法简单、快速、回收率高、重复性好,适用于动物源食品中(F)QNs的定量及确证检测。
A new method was developed for the determination of(fluoro)quinolones((F)QNs) residues in livestock and poultry meat using TurboFlow online cleanup(TFC)-liquid chromatography-tandem mass spectrometry(LC-MS/MS). Samples were extracted with methanol-acetonitrile(1 : 1, V/V) mixture. Then the extract was purified on a Cyclone-p column. Subsequently, the analytes were separated on a Hypersil Gold C_8 column before being analyzed by tandem mass spectrometry. The calibration curves showed good linearity with R~2 > 0.99 in the concentration range from 0.2 to 20.0 ng/mL for cinoxacin, enrofloxacin and danofloxacin, and in the range from 0.1 to 10.0 ng/mL for 17 other(F)QNs. The limit of quantization for enrofloxacin, danofloxacin and cinoxacin was 2 μg/kg, while that for 17 other(F)QNs was 1 μg/kg. Average recoveries of the analytes fortified at three levels ranged from 85.4% to 108.2%, with relative standard deviations(RSDs) between 4.04% and 8.57%. The method was characterized by simplicity, rapidity, high recovery and good repeatability and was suitable for quantitative analysis and confirmation of(F)QNs residues in foods of animal origin.
A new method was developed for the determination of(fluoro)quinolones((F)QNs) residues in livestock and poultry meat using TurboFlow online cleanup(TFC)-liquid chromatography-tandem mass spectrometry(LC-MS/MS). Samples were extracted with methanol-acetonitrile(1 : 1, V/V) mixture. Then the extract was purified on a Cyclone-p column. Subsequently, the analytes were separated on a Hypersil Gold C_8 column before being analyzed by tandem mass spectrometry. The calibration curves showed good linearity with R~2 > 0.99 in the concentration range from 0.2 to 20.0 ng/mL for cinoxacin, enrofloxacin and danofloxacin, and in the range from 0.1 to 10.0 ng/mL for 17 other(F)QNs. The limit of quantization for enrofloxacin, danofloxacin and cinoxacin was 2 μg/kg, while that for 17 other(F)QNs was 1 μg/kg. Average recoveries of the analytes fortified at three levels ranged from 85.4% to 108.2%, with relative standard deviations(RSDs) between 4.04% and 8.57%. The method was characterized by simplicity, rapidity, high recovery and good repeatability and was suitable for quantitative analysis and confirmation of(F)QNs residues in foods of animal origin.
引文
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[18]吴小莲,向垒,莫测辉,等.超高效液相色谱-电喷雾串联质谱测定蔬菜中喹诺酮类抗生素[J].分析化学,2013,41(6):876-881.DOI:10.3724/SP.J.1096.2013.20909.
[19]章红,易路遥,熊雯,等.Qu ECh ERS-四级杆飞行时间高分辨质谱法快速筛查典型水产品中的喹诺酮类药物残留[J].农产品质量与安全,2017(4):70-74.
[20]张中印,赵柳微,曹葳蕤,等.Qu ECh ERS前处理方法结合高效液相色谱-串联质谱测定蜂蜜和蜂王浆中14种喹诺酮类药物残留[J].食品科学,2016,37(16):242-248.DOI:10.7506/spkx1002-6630-201616039.
[21]李丽莉,罗轶,何颂华,等.高效液相色谱-串联质谱法测定鱼肉中8种喹诺酮类药物的残留量[J].中国食品卫生杂志,2012(1):37-40.
[22]刘芃岩,申杰,刘磊.复合模板印迹聚合物净化液相色谱-质谱联用法测定鱼肉中氟喹诺酮类残留[J].分析化学,2012,40(5):693-698.DOI:10.3724/SP.J.1096.2012.10610.
[23]杨盛茹,张恒业,张煌,等.UPLC-MS/MS检测鸡蛋中22种喹诺酮类药物残留方法的研究[J].现代食品科技,2017,33(8):262-268.DOI:10.13982/j.mfst.1673-9078.2017.8.038.
[24]艾连峰,马育松,陈瑞春,等.在线净化液相色谱串联质谱法测定动物源食品中金刚烷胺的残留[J].分析化学,2013,41(8):1194-1198.DOI:10.3724/SP.J.1096.2013.30047.
[25]MOTTIER P,HAMMEL Y A,GREMAUD E,et al.Quantitative high-throughput analysis of 16(fluoro)quinolones in honey using automated extraction by turbulent flow chromatography coupled to liquid chromatography-tandem mass spectrometry[J].Journal of Agricultural and Food Chemistry,2008,56(1):35-43.DOI:10.1021/jf072934d.
[26]ROACH J A G,DIBUSSOLO J M,KRYNITSKY A,et al.Evaluation and single laboratory validation of an on-line turbulent flow extraction tandem mass spectrometry method for melamine in infant formula[J].Journal of Chromatography A,2011,1218(1):4284-4290.DOI:10.1016/j.chroma.2011.05.025.
[27]赵孔祥,葛宝坤,陈旭艳,等.在线免疫亲和净化-液相色谱-串联质谱快速测定中草药及中成药中10种真菌毒素[J].分析化学,2011,39(9):1341-1346.DOI:10.3724/SP.J.1096.2011.01341.
[28]祝子铜,徐佳文,雷美康,等.HPLC-MS-MS同位素内标法测定火腿中14种喹诺酮类残留量[J].食品科学,2014,35(20):258-264.DOI:10.7506/spkx1002-6630-201420051.
[29]姚倩钰,贺德春,龚道新,等.高效液相色谱-串联质谱法测定废水中5种喹诺酮类抗生素[J].中国环境监测,2017,33(3):139-146.DOI:10.19316/j.issn.1002-6002.2017.03.21.
[30]SUYOUNG L,BYUNGJOO K,JEONGKWON K.Development of isotope dilution-liquid chromatography tandem mass spectrometry for the accurate determination of fluoroquinolones in animal meat products:optimization of chromatographic separation for eliminating matrix effects on isotope ratio measurements[J].Journal of Chromatography A,2013,1277(1):35-41.DOI:10.1016/j.chroma.2012.12.047.
[2]邱家学,李娜.浅析我国抗生素滥用现状及危害[J].上海医药,2006,27(12):555-556.DOI:10.3969/j.issn.1006-1533.2006.12.009.
[3]农业部.中华人民共和国农业部公告第235号动物性食品中兽药最高残留限量[EB/OL].(2002-12-24)[2018-01-07].http://jckspaqj.aqsiq.gov.cn/dwyxspjyjy/gnxgbz/200610/t20061027_9809.htm.
[4]European Union.Commision Regulation(EU)No 37/2010 of 22December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin[EB/OL].(2009-12-22)[2018-01-07].http://ec.europa.eu/health/files/mrl/mrl_20101212_consol.pdf.
[5]Electronic Code of Federal Regulations.PART 556:tolerances for residues of new animal drugs in food[EB/OL].(2013-09-05)[2018-01-07].http://www.ecfr.gov/cgi-bin/text-idx?c=ecfr&sid=407e94c4cd1bff327c7146f42bcc0d56&tpl=/ecfrbrowse/Title21/21cfr556_main_02.tpl.
[6]The Japan Food Chemical Research Foundation.Maximum residue limits(MRLs)list of agricultural chemicals in foods[EB/OL].(2013-08-06)[2018-01-07].http://www.m5.ws001.squarestart.ne.jp/foundation/search.html.
[7]Codex Alimentarius Commission.Index of veterinary drugs[EB/OL].[2018-01-07].http://www.fao.org/fao-who-codexalimentarius/codextexts/dbs/vetdrugs/veterinary-drugs/en/.
[8]VAN V G,JàNOSI A,BORDIN G,et al.Multiresidue determination of(fluoro)quinolone antibiotics in swine kidney using liquid c h r o m a t o g r a p h y-t a n d e m m a s s s p e c t r o m e t r y[J].J o u r n a l o f Chromatography A,2002,952(1):121-129.DOI:10.1016/S0021-9673(02)00092-4.
[9]TOUSSAINT B,BORDIN G,JANOSI A,et al.Validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of 11(fluoro)quinolone antibiotics in swine kidney[J].Journal of Chromatography A,2002,976(1):195-206.DOI:10.1016/S0021-9673(02)00948-2.
[10]CHEN Ligang,ZHANG Xiaopan,XU Yang,et al.Determination of fluoroquinolone antibiotics in environmental water samples based on magnetic molecularly imprinted polymer extraction followed by liquid chromatography-tandem mass spectrometry[J].Analytica Chimica Acta,2010,662(1):31-38.DOI:10.1016/j.aca.2010.01.001.
[11]马丽丽,郭昌胜,胡伟,等.固相萃取-高效液相色谱-串联质谱法同时测定土壤中氟喹诺酮、四环素和磺胺类抗生素[J].分析化学,2010,38(1):21-26.
[12]杨方,庞国芳,刘正才,等.液相色谱-串联质谱法检测水产品中15种喹诺酮类药物残留量[J].分析试验室,2008(12):27-33.DOI:10.3969/j.issn.1000-0720.2008.12.007.
[13]曹彦忠,庞国芳,张进杰,等.蜂蜜中14种喹诺酮类药物残留的高效液相色谱-串联质谱测定[J].分析测试学报,2008,27(11):1141-1146.DOI:10.3969/j.issn.1004-4957.2008.11.001.
[14]包晓丽,任一平,张虹.超高效液相色谱-电喷雾串联四极杆质谱法检测牛奶中22种喹诺酮类抗菌素[J].分析化学,2009,37(3):389-394.DOI:10.3321/j.issn:0253-3820.2009.03.015.
[15]何强,孔祥虹,李建华,等.超高效液相色谱-串联质谱法同时测定蜂蜜中硝基咪唑类、磺胺类、喹诺酮类兽药残留[J].分析试验室,2010(3):101-105.DOI:10.3969/j.issn.1000-0720.2010.08.015.
[16]孟哲,石志红,吕运开,等.超高效液相色谱-高分辨四级杆飞行时间质谱法快速筛查乳制品中磺胺类与氟喹诺酮类药物[J].分析化学,2014,42(10):1493-1500.DOI:10.11895/j.issn.0253-3820.140299.
[17]王金秋,马建民,夏曦,等.超高效液相色谱-串联质谱法同时测定猪肌肉中13种喹诺酮类药物残留[J].质谱学报,2014,35(2):185-192.DOI:10.7538/zpxb.youxian.2014.0003.
[18]吴小莲,向垒,莫测辉,等.超高效液相色谱-电喷雾串联质谱测定蔬菜中喹诺酮类抗生素[J].分析化学,2013,41(6):876-881.DOI:10.3724/SP.J.1096.2013.20909.
[19]章红,易路遥,熊雯,等.Qu ECh ERS-四级杆飞行时间高分辨质谱法快速筛查典型水产品中的喹诺酮类药物残留[J].农产品质量与安全,2017(4):70-74.
[20]张中印,赵柳微,曹葳蕤,等.Qu ECh ERS前处理方法结合高效液相色谱-串联质谱测定蜂蜜和蜂王浆中14种喹诺酮类药物残留[J].食品科学,2016,37(16):242-248.DOI:10.7506/spkx1002-6630-201616039.
[21]李丽莉,罗轶,何颂华,等.高效液相色谱-串联质谱法测定鱼肉中8种喹诺酮类药物的残留量[J].中国食品卫生杂志,2012(1):37-40.
[22]刘芃岩,申杰,刘磊.复合模板印迹聚合物净化液相色谱-质谱联用法测定鱼肉中氟喹诺酮类残留[J].分析化学,2012,40(5):693-698.DOI:10.3724/SP.J.1096.2012.10610.
[23]杨盛茹,张恒业,张煌,等.UPLC-MS/MS检测鸡蛋中22种喹诺酮类药物残留方法的研究[J].现代食品科技,2017,33(8):262-268.DOI:10.13982/j.mfst.1673-9078.2017.8.038.
[24]艾连峰,马育松,陈瑞春,等.在线净化液相色谱串联质谱法测定动物源食品中金刚烷胺的残留[J].分析化学,2013,41(8):1194-1198.DOI:10.3724/SP.J.1096.2013.30047.
[25]MOTTIER P,HAMMEL Y A,GREMAUD E,et al.Quantitative high-throughput analysis of 16(fluoro)quinolones in honey using automated extraction by turbulent flow chromatography coupled to liquid chromatography-tandem mass spectrometry[J].Journal of Agricultural and Food Chemistry,2008,56(1):35-43.DOI:10.1021/jf072934d.
[26]ROACH J A G,DIBUSSOLO J M,KRYNITSKY A,et al.Evaluation and single laboratory validation of an on-line turbulent flow extraction tandem mass spectrometry method for melamine in infant formula[J].Journal of Chromatography A,2011,1218(1):4284-4290.DOI:10.1016/j.chroma.2011.05.025.
[27]赵孔祥,葛宝坤,陈旭艳,等.在线免疫亲和净化-液相色谱-串联质谱快速测定中草药及中成药中10种真菌毒素[J].分析化学,2011,39(9):1341-1346.DOI:10.3724/SP.J.1096.2011.01341.
[28]祝子铜,徐佳文,雷美康,等.HPLC-MS-MS同位素内标法测定火腿中14种喹诺酮类残留量[J].食品科学,2014,35(20):258-264.DOI:10.7506/spkx1002-6630-201420051.
[29]姚倩钰,贺德春,龚道新,等.高效液相色谱-串联质谱法测定废水中5种喹诺酮类抗生素[J].中国环境监测,2017,33(3):139-146.DOI:10.19316/j.issn.1002-6002.2017.03.21.
[30]SUYOUNG L,BYUNGJOO K,JEONGKWON K.Development of isotope dilution-liquid chromatography tandem mass spectrometry for the accurate determination of fluoroquinolones in animal meat products:optimization of chromatographic separation for eliminating matrix effects on isotope ratio measurements[J].Journal of Chromatography A,2013,1277(1):35-41.DOI:10.1016/j.chroma.2012.12.047.