高效液相色谱-串联飞行时间质谱高通量筛查生鲜乳中兽药残留
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摘要
建立了一种高通量检测生鲜乳中兽药残留的高效液相色谱-串联飞行时间质谱方法,包括17大类兽药,共174种。样品用乙腈和乙酸乙酯提取,冷冻净化后,目标化合物经过Poroshell120EC-C18柱(3.0 mm×150 mm,2.7μm)分离,经0.1%甲酸溶液和甲醇流动相洗脱,采用电喷雾正离子模式进行采集。结果显示,114种兽药化合物在质量浓度0～200 ng/mL范围线性良好(R2>0.990 0),检测限为1～10 ng/g。此外,回收率在70%～130%范围内的质控加标样品达99种,相对标准偏差为1.03%～9.65%。该方法检测速度快、灵敏度高,大大提高了生鲜乳中兽药筛查的效率。
In this study, a high performance liquid chromatography tandem time-of-flight mass spectrometry(HPLC-TOFMS/MS) method was established for the high-throughput determination of 174 veterinary drug residues in raw milk. The target compounds were extracted with acetonitrile and purified by freezing. A Poroshell 120 EC-C18 column(3.0 mm × 150 mm, 2.7 μm) was used for the separation of the target compounds with a mobile phase consisting of 0.1% formic acid in water and methanol via gradient elution. The analytes were detected with an electrospray ionization source(EIS) in the positive ion mode. The results showed that the method had a good linearity(R2 > 0.990 0) and low limits of detection(LOD)(1–10 ng/g) for 114 pesticide compounds. The average recoveries for 99 of these ranged from 70% to 130% with relative standard deviation(RSD) from 1.03% to 9.65% at quality control concentration levels. The method was successfully applied in the screening of veterinary drug residues in raw milk with high efficiency and sensitivity.
In this study, a high performance liquid chromatography tandem time-of-flight mass spectrometry(HPLC-TOFMS/MS) method was established for the high-throughput determination of 174 veterinary drug residues in raw milk. The target compounds were extracted with acetonitrile and purified by freezing. A Poroshell 120 EC-C18 column(3.0 mm × 150 mm, 2.7 μm) was used for the separation of the target compounds with a mobile phase consisting of 0.1% formic acid in water and methanol via gradient elution. The analytes were detected with an electrospray ionization source(EIS) in the positive ion mode. The results showed that the method had a good linearity(R2 > 0.990 0) and low limits of detection(LOD)(1–10 ng/g) for 114 pesticide compounds. The average recoveries for 99 of these ranged from 70% to 130% with relative standard deviation(RSD) from 1.03% to 9.65% at quality control concentration levels. The method was successfully applied in the screening of veterinary drug residues in raw milk with high efficiency and sensitivity.
引文
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[6]王鹏思,李嘉欣,石上梅,等.GC法测定市售动物源性中药材地龙、水蛭中3种氯霉素类药物的残留量[J].中国兽药杂志,2017,51(3):65-69.
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[10]TAO Y,CHEN D,YU G,et al.Simultaneous determination of lincomycin and spectinomycin residues in animal tissues by gas chromatography-nitrogen phosphorus detection and gas chromatography-mass spectrometry with accelerated solvent extraction[J].Food Additives&Contaminants,2011,28(2):145-154.DOI:10.1080/19440049.2010.538440.
[11]YIKILMAZ Y,FILAZI A.Detection of florfenicol residues in salmon trout via GC-MS[J].Food Analytical Methods,2015,8(4):1-7.DOI:10.1007/s12161-014-9982-8.
[12]YAMAGUCHI T,KAKIMOTO K,NAGAYOSHIH,et al.Simultaneous determination of veterinary drugs in livestock products using dispersive and cartridge column solid-phase extraction by LC-MS/MS[J].Journal of the Food Hygienic Society of Japan,2013,54(4):290-297.DOI:10.3358/shokueishi.54.290.
[13]KIM S C,CARLSON K.Quantification of human and veterinary antibiotics in water and sediment using SPE/LC/MS/MS[J].Analytical&Bioanalytical Chemistry,2007,387(4):1301-1315.DOI:10.1007/s00216-006-0613-0.
[14]CHEN X H,ZHAO Y G,QIU Q L,et al.A fast and high throughput LC-MS/MS method for the determination of 58 human and veterinary drugs in river water[J].Analytical Methods,2017,9(29):4228-4233.DOI:10.1039/c7ay01240c.
[15]DENG X,ZHU L,FANG T,et al.Analysis of isoquinoline alkaloid composition and wound-induced variation in nelumbo using HPLC-MS/MS[J].Journal of Agricultural&Food Chemistry,2016,64(5):1130-1136.DOI:10.1021/acs.jafc.5b06099.
[16]SHENDY A H,AL-GHOBASHY M A,GAD AlLLA S A,et al.Development and validation of a modified QuEChERS protocol coupled to LC-MS/MS for simultaneous determination of multi-class antibiotic residues in honey[J].Food Chemistry,2016,190:982-989.DOI:10.1016/j.foodchem.2015.06.048.
[17]HSU B Y,LIN S W,INBARAJ B S,et al.Simultaneous determination of phenolic acids and flavonoids in Chenopodium formosanum Koidz.(djulis)by HPLC-DAD-ESI-MS/MS[J].Journal of Pharmaceutical and Biomedical Analysis,2016,132:109-116.DOI:10.1002/jssc.201401008.
[18]张红印,马丽卿.气相色谱法与液相色谱法的比较及应用[J].肉类工业,2000(10):38-39.DOI:10.3969/j.issn.1008-5467.2000.10.013.
[19]QIANMC,PETERSOND G,REINECCIUSG A.Gas chromatography[M].Springer International Publishing,2017:155-164.
[20]姜凤丽,滕小梅,倪春芳,等.超高效液相色谱-串联四极杆飞行时间质谱法筛查人全血中150种药物与毒物[J].理化检验:化学分册,2016,52(4):417-426.DOI:10.11973/1hjy-hx201604010.
[21]LI J Z.Multicalss residues screening of 105 veterinary drugs in meat,milk,and egg using ultra high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry[J].Journal of Liquid Chromatography&Related Technologies,2010,34(19):2286-2303.DOI:10.1080/10826076.2011.587224.
[22]MARTIN J,BUCHBERGER W,ALONSO E,et al.Comparison of different extraction methods for the determination of statin drugs in wastewater and river water by HPLC/Q-TOF-MS[J].Talanta,2011,85(1):607-615.DOI:10.1016/j.talanta.2011.04.017.
[23]GU J F,FENG L,ZHANG M H,et al.New metabolite profiles of Danshensu in rats by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry[J].Journal of Chromatography B,2014,955/956(1):20-25.DOI:10.1016/j.jchromb.2014.02.010.
[24]SONG H,WANG J,ZHANG R,et al.In vivo metabolism study of bergenin in rats by HPLC-QTOF mass spectrometry[J].Biomedical Chromatography,2013,27(11):1398-1405.DOI:10.1002/bmc.2934.
[25]孙清荣,郭礼强,张金玲,等.HPLC-Q-TOF法筛查鸡肉中48种兽药残留[J].食品研究与开发,2017,38(4):127-132.DOI:10.3969/j.issn.1005-6521.2017.04.027.
[26]朱万燕,徐文远,杨晓玉.食品中多种兽药残留的同时筛查检测[J].科技创新导报,2017(4):51-53.DOI:10.16660/j.cnki.1674-098X.2017.04.051.
[27]朱万燕,徐文远,张伟.超高效液相色谱-四级杆-飞行时间质谱法同时快速检测水产品中37种兽药残留[J].食品安全质量检测学报,2017,8(2):614-619.DOI:10.3969/j.issn.2095-0381.2017.02.041.
[28]高馥蝶,赵妍,邵兵,等.超高效液相色谱-四极杆-飞行时间质谱法快速筛查牛奶中的农药和兽药残留[J].色谱,2012,30(6):560-567.DOI:10.3724/SP.J.1123.2012.02021.
[29]杨路平,焦燕妮,邵立君,等.超高效液相色谱-质谱法测定饮用水中14种氟喹诺酮类抗生素的残留量[J].理化检验:化学分册,2016,52(4):397-399.DOI:10.3724/SP.J.1123.2012.02021.
[30]ROBERT C,GILLARD N,BRASSEUR P Y,et al.Rapid multi-residue and multi-class qualitative screening for veterinary drugs in foods of animal origin by UHPLC-MS/MS[J].Food Additives&Contaminants,2013,30(3):443-457.DOI:10.1080/19440049.2012.751632.
[2]农业部.中华人民共和国农业部公告第235号[S].北京:中国农业出版社,2002.
[3]李明,董春柳.浅谈动物性食品中兽药残留的现状及防控策略[J].现代畜牧科技,2016(6):172.DOI:10.3969/j.issn.1673-1921.2016.06.164.
[4]JIANG T F,WANG Y H,LV Z H,et al.Direct determination of barbiturates in urine by capillary electrophoresis using a capillary coated dynamically with polycationic polymers[J].Chromatographia,2007,65(9/10):611-615.DOI:10.1365/s10337-007-0213-y.
[5]LOPEZ A V,YOUNG R,BECKERT W F.On-line determination of organophosphorus pesticides in water by solid-phase microextraction and gas chromatography with thermionic-selective detection[J].Journal of High Resolution Chromatography,2015,20(9):487-492.DOI:10.1002/jhrc.1240200905.
[6]王鹏思,李嘉欣,石上梅,等.GC法测定市售动物源性中药材地龙、水蛭中3种氯霉素类药物的残留量[J].中国兽药杂志,2017,51(3):65-69.
[7]MASIA A,SUAREZVARELA M M,LLOPISONZALEZ A,et al.Determination of pesticides and veterinary drug residues in food by liquid chromatography-mass spectrometry:a review[J].Analytica Chimica Acta,2016,936:40-61.DOI:10.1016/j.aca.2016.07.023.
[8]GUO P,CHEN Y,YUE C,et al.Simultaneous determination of clavulanic acid and tazobactam in bovine milk by HPLC[J].Food Additives&Contaminents:Part A,2017,34(4):617-623.DOI:10.1080/19440049.2016.1277271.
[9]CHANG G R,CHEN H S,LIN F Y.Analysis of banned veterinary drugs and herbicide residues in shellfish by liquid chromatographytandem mass spectrometry(LC/MS/MS)and gas chromatographytandem mass spectrometry(GC/MS/MS)[J].Marine Pollution Bulletin,2016,113:579-584.DOI:10.1016/j.marpolbul.2016.08.080.
[10]TAO Y,CHEN D,YU G,et al.Simultaneous determination of lincomycin and spectinomycin residues in animal tissues by gas chromatography-nitrogen phosphorus detection and gas chromatography-mass spectrometry with accelerated solvent extraction[J].Food Additives&Contaminants,2011,28(2):145-154.DOI:10.1080/19440049.2010.538440.
[11]YIKILMAZ Y,FILAZI A.Detection of florfenicol residues in salmon trout via GC-MS[J].Food Analytical Methods,2015,8(4):1-7.DOI:10.1007/s12161-014-9982-8.
[12]YAMAGUCHI T,KAKIMOTO K,NAGAYOSHIH,et al.Simultaneous determination of veterinary drugs in livestock products using dispersive and cartridge column solid-phase extraction by LC-MS/MS[J].Journal of the Food Hygienic Society of Japan,2013,54(4):290-297.DOI:10.3358/shokueishi.54.290.
[13]KIM S C,CARLSON K.Quantification of human and veterinary antibiotics in water and sediment using SPE/LC/MS/MS[J].Analytical&Bioanalytical Chemistry,2007,387(4):1301-1315.DOI:10.1007/s00216-006-0613-0.
[14]CHEN X H,ZHAO Y G,QIU Q L,et al.A fast and high throughput LC-MS/MS method for the determination of 58 human and veterinary drugs in river water[J].Analytical Methods,2017,9(29):4228-4233.DOI:10.1039/c7ay01240c.
[15]DENG X,ZHU L,FANG T,et al.Analysis of isoquinoline alkaloid composition and wound-induced variation in nelumbo using HPLC-MS/MS[J].Journal of Agricultural&Food Chemistry,2016,64(5):1130-1136.DOI:10.1021/acs.jafc.5b06099.
[16]SHENDY A H,AL-GHOBASHY M A,GAD AlLLA S A,et al.Development and validation of a modified QuEChERS protocol coupled to LC-MS/MS for simultaneous determination of multi-class antibiotic residues in honey[J].Food Chemistry,2016,190:982-989.DOI:10.1016/j.foodchem.2015.06.048.
[17]HSU B Y,LIN S W,INBARAJ B S,et al.Simultaneous determination of phenolic acids and flavonoids in Chenopodium formosanum Koidz.(djulis)by HPLC-DAD-ESI-MS/MS[J].Journal of Pharmaceutical and Biomedical Analysis,2016,132:109-116.DOI:10.1002/jssc.201401008.
[18]张红印,马丽卿.气相色谱法与液相色谱法的比较及应用[J].肉类工业,2000(10):38-39.DOI:10.3969/j.issn.1008-5467.2000.10.013.
[19]QIANMC,PETERSOND G,REINECCIUSG A.Gas chromatography[M].Springer International Publishing,2017:155-164.
[20]姜凤丽,滕小梅,倪春芳,等.超高效液相色谱-串联四极杆飞行时间质谱法筛查人全血中150种药物与毒物[J].理化检验:化学分册,2016,52(4):417-426.DOI:10.11973/1hjy-hx201604010.
[21]LI J Z.Multicalss residues screening of 105 veterinary drugs in meat,milk,and egg using ultra high performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry[J].Journal of Liquid Chromatography&Related Technologies,2010,34(19):2286-2303.DOI:10.1080/10826076.2011.587224.
[22]MARTIN J,BUCHBERGER W,ALONSO E,et al.Comparison of different extraction methods for the determination of statin drugs in wastewater and river water by HPLC/Q-TOF-MS[J].Talanta,2011,85(1):607-615.DOI:10.1016/j.talanta.2011.04.017.
[23]GU J F,FENG L,ZHANG M H,et al.New metabolite profiles of Danshensu in rats by ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry[J].Journal of Chromatography B,2014,955/956(1):20-25.DOI:10.1016/j.jchromb.2014.02.010.
[24]SONG H,WANG J,ZHANG R,et al.In vivo metabolism study of bergenin in rats by HPLC-QTOF mass spectrometry[J].Biomedical Chromatography,2013,27(11):1398-1405.DOI:10.1002/bmc.2934.
[25]孙清荣,郭礼强,张金玲,等.HPLC-Q-TOF法筛查鸡肉中48种兽药残留[J].食品研究与开发,2017,38(4):127-132.DOI:10.3969/j.issn.1005-6521.2017.04.027.
[26]朱万燕,徐文远,杨晓玉.食品中多种兽药残留的同时筛查检测[J].科技创新导报,2017(4):51-53.DOI:10.16660/j.cnki.1674-098X.2017.04.051.
[27]朱万燕,徐文远,张伟.超高效液相色谱-四级杆-飞行时间质谱法同时快速检测水产品中37种兽药残留[J].食品安全质量检测学报,2017,8(2):614-619.DOI:10.3969/j.issn.2095-0381.2017.02.041.
[28]高馥蝶,赵妍,邵兵,等.超高效液相色谱-四极杆-飞行时间质谱法快速筛查牛奶中的农药和兽药残留[J].色谱,2012,30(6):560-567.DOI:10.3724/SP.J.1123.2012.02021.
[29]杨路平,焦燕妮,邵立君,等.超高效液相色谱-质谱法测定饮用水中14种氟喹诺酮类抗生素的残留量[J].理化检验:化学分册,2016,52(4):397-399.DOI:10.3724/SP.J.1123.2012.02021.
[30]ROBERT C,GILLARD N,BRASSEUR P Y,et al.Rapid multi-residue and multi-class qualitative screening for veterinary drugs in foods of animal origin by UHPLC-MS/MS[J].Food Additives&Contaminants,2013,30(3):443-457.DOI:10.1080/19440049.2012.751632.