鸽蛋中氯霉素甲砜霉素氟苯尼考及其代谢物氟苯尼考胺同时检测的超高效液相色谱-串联质谱法的建立
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摘要
为建立同时检测鸽蛋中氯霉素(CAP)、甲砜霉素(TAP)、氟苯尼考(FF)及其代谢物氟苯尼考胺(FFA)多残留的超高效液相色谱-串联质谱(UPLC-MS/MS)法,将样品经过液-液萃取(LLE)或者加速溶剂萃取(ASE)提取、净化,采用电喷雾离子源(ESI)模式和多反应监测模式(MRM),通过UPLC-MS/MS检测和确证分析。结果显示,CAP、TAP、FF和FFA添加水平分别为定量限(LOQ),0.5、1.0和2.0倍的最低要求执行限量(MRPL)或最高残留限量(MRL)时,平均回收率在91.55%～105.47%范围内,相对标准偏差(RSD)均低于3.04%,日内RSD为1.83%～3.70%,日间RSD为2.72%～4.62%。CAP、TAP、FF和FFA在鸽蛋中的检测限(LODs)为0.03～0.4ug/kg(S/N≥3),LOQs为0.08～1.2ug/kg(S/N≥10),其显著低于最大残留限量(MRL)。结果表明,此方法快速、准确、灵敏度高,定性、定量准确,适用于鸽蛋中氯霉素、甲砜霉素、氟苯尼考及其代谢物氟苯尼考胺多残留的检测。
A widely applicable method involving ultra-performance liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry(UPLC-ESI/MS/MS) was developed for the simultaneous determination of chloramphenicol(CAP),thiamphenicol(TAP),florfenicol(FF) and florfenicol amine(FFA) in pigeon eggs.Samples was extracted and purified by liquid-liquid extraction(LLE) or accelerated solvent extraction(ASE),using ESI mode and multiple reaction monitoring mode(MRM) by UPLC-MS/MS detection and confirmation analysis.When the added concentration of CAP,TAP,FF and FFA were the limit of quantification(LOQ),0.5,1.0 and 2.0 times of the minimum required performance limit(MRPL) or maximum residue limit(MRL),respectively,the average recoveries of the four analytes from pigeon egg samples were from 91.55% to 105.47%,with relative standard deviations(RSDs)of less than 3.04%.The intra-day RSD values were from 1.83% to 3.70%,and the inter-day RSD values were from 2.72% to 4.62%.Limits of detection(LODs) were from 0.03 to 0.4 ug/kg(S/N≥3) for CAP,TAP,FF and FFA in pigeon eggs,and LOQs were from 0.08 to 1.2 ug/kg(S/N≥10),these values were significantly below the maximum residue limit(MRL).In conclusion,the method is rapid,accurate,sensitive,accurate qualitative and quantitative,which is suitable for multi-residue detection of chloramphenicol,thiamphenicol,florfenicol and its metabolite florfenicol amine in pigeon eggs.
A widely applicable method involving ultra-performance liquid chromatography-electrospray ionization tandem triple quadrupole mass spectrometry(UPLC-ESI/MS/MS) was developed for the simultaneous determination of chloramphenicol(CAP),thiamphenicol(TAP),florfenicol(FF) and florfenicol amine(FFA) in pigeon eggs.Samples was extracted and purified by liquid-liquid extraction(LLE) or accelerated solvent extraction(ASE),using ESI mode and multiple reaction monitoring mode(MRM) by UPLC-MS/MS detection and confirmation analysis.When the added concentration of CAP,TAP,FF and FFA were the limit of quantification(LOQ),0.5,1.0 and 2.0 times of the minimum required performance limit(MRPL) or maximum residue limit(MRL),respectively,the average recoveries of the four analytes from pigeon egg samples were from 91.55% to 105.47%,with relative standard deviations(RSDs)of less than 3.04%.The intra-day RSD values were from 1.83% to 3.70%,and the inter-day RSD values were from 2.72% to 4.62%.Limits of detection(LODs) were from 0.03 to 0.4 ug/kg(S/N≥3) for CAP,TAP,FF and FFA in pigeon eggs,and LOQs were from 0.08 to 1.2 ug/kg(S/N≥10),these values were significantly below the maximum residue limit(MRL).In conclusion,the method is rapid,accurate,sensitive,accurate qualitative and quantitative,which is suitable for multi-residue detection of chloramphenicol,thiamphenicol,florfenicol and its metabolite florfenicol amine in pigeon eggs.
引文
[1]中华人民共和国农业部.动物性食品中兽药最高残留限量[S]//公告第235号(附录4).北京:农业部,2002.Ministry of Agriculture of the People’s Republic of China.Maximum residue level of veterinary drugs in food of animal origin[S]//Notice No.235(Appendix 4).Beijing:Ministry of Agriculture,2002.(in Chinese)
[2]U.S.Food and Drug Administration.CFR-code of federal regulations title 21 part 556 tolerances for residue of new animal drugs in food[S].2014.
[3]The European Medicines Agency.COMMISSION REGULA-TION(EU)No.37/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin[S].2010.
[4]赵霞,王波,谢恺舟,等.动物性食品中氯霉素类药物残留色谱和质谱检测技术研究进展[J].分析试验室,2018,37(8):978-985.ZHAO Xia,WANG Bo,XIE Kai-zhou,et al.An overview on chromatographic and mass spectrometry analytical methods for the detection of chloramphenicols residues in animal food[J].Chinese Journal of Analysis Laboratory,2018,37(8):978-985.(in Chinese)
[5]XIE K,JIA L,YAO Y,et al.Simultaneous determination of thiamphenicol,florfenicol and florfenicol amine in eggs by reversed-phase high-performance liquid chromatography with fluorescence detection[J].Journal of Chromatography B,2011,879(23):2351-2354.
[6]ZHANG S,LIU Z,GUO X,et al.Simultaneous determination and confirmation of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine in chicken muscle by liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography B,2008,875(2):399-404.
[7]BARRETO F,RIBEIRO C,HOFF R B,et al.Determination of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine in poultry,swine,bovine and fish by liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2016,1449:48-53.
[8]FEDENIUK R W,MIZUNO M,NEISER C,et al.Development of LC-MS/MS methodology for the detection/determination and confirmation of chloramphenicol,chloramphenicol 3-O-β-d-glucuronide,florfenicol,florfenicol amine and thiamphenicol residues in bovine,equine and porcine liver[J].Journal of Chromatography B,2015,991:68-78.
[9]XIE X,WANG B,PANG M,et al.Quantitative analysis of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine in eggs via liquid chromatography-electrospray ionization tandem mass spectrometry[J].Food Chemistry,2018,269:542-548.
[10]ORLANDO E A,ROQUE A G C,LOSEKANN M E,et al.UPLC-MS/MS determination of florfenicol and florfenicol amine antimicrobial residues in tilapia muscle[J].Journal of Chromatography B,2016,1035:8-15.
[11]XIAO Z,SONG R,RAO Z,et al.Development of a subcritical water extraction approach for trace analysis of chloramphenicol,thiamphenicol,florfenicol,and florfenicol amine in poultry tissues[J].Journal of Chromatography A,2015,1418(4):29-35.
[12]贾宏新,邹淼,林盛.超高效液相色谱-串联质谱法测定河蟹中氯霉素类药物残留[J].中国卫生检验杂志,2013,25(13):2726-2729.JIA Hong-xin,ZHOU Miao,LIN Sheng.Determination of chloramphenicol,thiamphnicol,florfenicol residues in river crab by UPLC-MS/MS[J].Chinese Journal of Health Laboratory Technology,2013,25(13):2726-2729.(in Chinese)
[13]潘晓东,吴平谷,姜维.超高效液相色谱-串联质谱法同时测定鱼肉中氯霉素、甲砜霉素和氟甲砜霉素[J].中国食品卫生杂志,2014,26(6):572-574.PAN Xiao-dong,WU Ping-gu,JIANG Wei.Determination of residues of chloramphenicol,thiamphenicol and florfenicol in fi sh muscle by ultra-performance liquid chromatography-tandem mass spectrometry[J].Chinese Journal of Food Hygiene,2014,26(6):572-574.(in Chinese)
[14]陈蔷,宋志超,张崇威,等.动物源食品中酰胺醇类药物及其代谢物残留检测超高效液相色谱-串联质谱法研究[J].中国兽药杂志,2015,49(8):28-34.CHEN Qiang,SONG Zhi-chao,ZHANG Chong-wei,et al.Determination of amphenicols and metabolite residues in animal derived food by UPLC-MS/MS[J].Chinese Journal of Veterinary Drug,2015,49(8):28-34.(in Chinese)
[15]杨洁,杨钊,于宙,等.超高效液相色谱-串联质谱法测定禽蛋中氯霉素、甲砜霉素和氟甲砜霉素[J].分析试验室,2016,35(12):1463-1467.YANG Jie,YANG Zhao,YU Zhou,et al.Determination of chloramphenicol,thiamphenicol and florfenicol in poultry eggs by ultra performance liquid chromatography-tandem mass spectrometry[J].Chinese Journal of Analysis Laboratory,2016,35(12):1463-1467.(in Chinese)
[16]曾勇,董文婷,龚波,等.超高效液相色谱-串联质谱法快速测定禽蛋中氯霉素、氟苯尼考及其代谢物残留[J].中国兽药杂志,2017,51(12):42-46.ZENG Yong,DONG Wen-ting,GONG Bo,et al.Rapid determination of chloramphenicol,florfenicol and its metabolites in poultry eggs by UPLC-MS/MS[J].Chinese Journal of Veterinary Drug,2017,51(12):42-46.(in Chinese)
[17]LI P,QIU Y,CAI H,et al.Simultaneous determination of chloramphenicol,thiamphenicol,and florfenicol residues in animal tissues by gas chromatography/mass spectrometry[J].Chinese Journal of Chromatography,2006,24(1):14-18.
[18]SHEN J,XIA X,JIANG H,et al.Determination of chloramphenicol,thiamphenicol,florfenicol,and florfenicol amine in poultry and porcine muscle and liver by gas chromatography-negative chemical ionization mass spectrometry[J].Journal of Chromatography B,2009,877(14-15):1523-1529.
[2]U.S.Food and Drug Administration.CFR-code of federal regulations title 21 part 556 tolerances for residue of new animal drugs in food[S].2014.
[3]The European Medicines Agency.COMMISSION REGULA-TION(EU)No.37/2010 of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin[S].2010.
[4]赵霞,王波,谢恺舟,等.动物性食品中氯霉素类药物残留色谱和质谱检测技术研究进展[J].分析试验室,2018,37(8):978-985.ZHAO Xia,WANG Bo,XIE Kai-zhou,et al.An overview on chromatographic and mass spectrometry analytical methods for the detection of chloramphenicols residues in animal food[J].Chinese Journal of Analysis Laboratory,2018,37(8):978-985.(in Chinese)
[5]XIE K,JIA L,YAO Y,et al.Simultaneous determination of thiamphenicol,florfenicol and florfenicol amine in eggs by reversed-phase high-performance liquid chromatography with fluorescence detection[J].Journal of Chromatography B,2011,879(23):2351-2354.
[6]ZHANG S,LIU Z,GUO X,et al.Simultaneous determination and confirmation of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine in chicken muscle by liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography B,2008,875(2):399-404.
[7]BARRETO F,RIBEIRO C,HOFF R B,et al.Determination of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine in poultry,swine,bovine and fish by liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2016,1449:48-53.
[8]FEDENIUK R W,MIZUNO M,NEISER C,et al.Development of LC-MS/MS methodology for the detection/determination and confirmation of chloramphenicol,chloramphenicol 3-O-β-d-glucuronide,florfenicol,florfenicol amine and thiamphenicol residues in bovine,equine and porcine liver[J].Journal of Chromatography B,2015,991:68-78.
[9]XIE X,WANG B,PANG M,et al.Quantitative analysis of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine in eggs via liquid chromatography-electrospray ionization tandem mass spectrometry[J].Food Chemistry,2018,269:542-548.
[10]ORLANDO E A,ROQUE A G C,LOSEKANN M E,et al.UPLC-MS/MS determination of florfenicol and florfenicol amine antimicrobial residues in tilapia muscle[J].Journal of Chromatography B,2016,1035:8-15.
[11]XIAO Z,SONG R,RAO Z,et al.Development of a subcritical water extraction approach for trace analysis of chloramphenicol,thiamphenicol,florfenicol,and florfenicol amine in poultry tissues[J].Journal of Chromatography A,2015,1418(4):29-35.
[12]贾宏新,邹淼,林盛.超高效液相色谱-串联质谱法测定河蟹中氯霉素类药物残留[J].中国卫生检验杂志,2013,25(13):2726-2729.JIA Hong-xin,ZHOU Miao,LIN Sheng.Determination of chloramphenicol,thiamphnicol,florfenicol residues in river crab by UPLC-MS/MS[J].Chinese Journal of Health Laboratory Technology,2013,25(13):2726-2729.(in Chinese)
[13]潘晓东,吴平谷,姜维.超高效液相色谱-串联质谱法同时测定鱼肉中氯霉素、甲砜霉素和氟甲砜霉素[J].中国食品卫生杂志,2014,26(6):572-574.PAN Xiao-dong,WU Ping-gu,JIANG Wei.Determination of residues of chloramphenicol,thiamphenicol and florfenicol in fi sh muscle by ultra-performance liquid chromatography-tandem mass spectrometry[J].Chinese Journal of Food Hygiene,2014,26(6):572-574.(in Chinese)
[14]陈蔷,宋志超,张崇威,等.动物源食品中酰胺醇类药物及其代谢物残留检测超高效液相色谱-串联质谱法研究[J].中国兽药杂志,2015,49(8):28-34.CHEN Qiang,SONG Zhi-chao,ZHANG Chong-wei,et al.Determination of amphenicols and metabolite residues in animal derived food by UPLC-MS/MS[J].Chinese Journal of Veterinary Drug,2015,49(8):28-34.(in Chinese)
[15]杨洁,杨钊,于宙,等.超高效液相色谱-串联质谱法测定禽蛋中氯霉素、甲砜霉素和氟甲砜霉素[J].分析试验室,2016,35(12):1463-1467.YANG Jie,YANG Zhao,YU Zhou,et al.Determination of chloramphenicol,thiamphenicol and florfenicol in poultry eggs by ultra performance liquid chromatography-tandem mass spectrometry[J].Chinese Journal of Analysis Laboratory,2016,35(12):1463-1467.(in Chinese)
[16]曾勇,董文婷,龚波,等.超高效液相色谱-串联质谱法快速测定禽蛋中氯霉素、氟苯尼考及其代谢物残留[J].中国兽药杂志,2017,51(12):42-46.ZENG Yong,DONG Wen-ting,GONG Bo,et al.Rapid determination of chloramphenicol,florfenicol and its metabolites in poultry eggs by UPLC-MS/MS[J].Chinese Journal of Veterinary Drug,2017,51(12):42-46.(in Chinese)
[17]LI P,QIU Y,CAI H,et al.Simultaneous determination of chloramphenicol,thiamphenicol,and florfenicol residues in animal tissues by gas chromatography/mass spectrometry[J].Chinese Journal of Chromatography,2006,24(1):14-18.
[18]SHEN J,XIA X,JIANG H,et al.Determination of chloramphenicol,thiamphenicol,florfenicol,and florfenicol amine in poultry and porcine muscle and liver by gas chromatography-negative chemical ionization mass spectrometry[J].Journal of Chromatography B,2009,877(14-15):1523-1529.