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
|
摘要
目的建立超高效液相色谱-四极杆-飞行时间质谱法快速检测水产品中磺胺类、喹诺酮类、四环素类、硝基咪唑类、大环内酯类、β-内酰胺类、三苯甲烷类7类37种兽药残留的方法。方法样品采用Qu ECh ERS方法(quick,easy,cheap,effective,rugged and safe method)进行前处理(用乙腈提取,提取液经Al_2O_3吸附剂净化);采用ZORBAX SB-C_(18)色谱柱(100 mm×2.1 mm,3.5μm)分离,以乙腈-5 mmol的乙酸铵为流动相,梯度洗脱,Q-TOF-MS电喷雾正离子模式分析检测;在全扫描采集模式下,以准分子离子峰的峰面积定量,以化合物的色谱保留时间和精确质量数定性。结果各化合物在0.5~2000μg/kg范围内均呈现良好的线性关系,相关系数均大于0.99。样品平均添加回收率为67%~109.5%,相对标准偏差(RSD)均小于15.3%(n=6)。结论方法简单、快速、可靠,可用于水产品中兽药多残留的快速筛查。
Objective To establish a method for the simultaneous determination of 7 classes of 37 kinds of veterinary drugs, including sulfoamides, quinolones, tetracyclines, nitroimidazoles, macrolides, beta-lactams and triphenylmethane, in aquatic products by ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS). Methods The quick, easy, cheap, effective, rugged and safe method(Qu ECh ERS) was used for the extraction. In this Qu ECh ERS method, the sample was extracted by acetonitrile and cleaned up with Al_2O_3. The compounds were measured on a ZORBAX SB-C_(18) column(100 mm×2.1 mm, 3.5 μm) with acetonitrile- 5 mmol/L ammonium acetate as mobile phase under gradient elution. The extract was measured directly by LC-Q TOF-MS with electrospray ionization in positive mode. The confirmatory analysis was carried out by determining the retention time and accurate masses of all compounds and fragment ions upon target MS/MS. Results The results showed that the linear range was 0.5~2000 μg/kg for each compound with the correlation coefficients of more than 0.99. The recoveries were ranged from 67.0% to 109.5%, with relative standard deviations(RSDs) less than 15.3%(n=6). Conclusion The method is simple, rapid and reliable, and is suitable for the simultaneous determination of multiresidues of veterinary drugs in aquatic products.
Objective To establish a method for the simultaneous determination of 7 classes of 37 kinds of veterinary drugs, including sulfoamides, quinolones, tetracyclines, nitroimidazoles, macrolides, beta-lactams and triphenylmethane, in aquatic products by ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS). Methods The quick, easy, cheap, effective, rugged and safe method(Qu ECh ERS) was used for the extraction. In this Qu ECh ERS method, the sample was extracted by acetonitrile and cleaned up with Al_2O_3. The compounds were measured on a ZORBAX SB-C_(18) column(100 mm×2.1 mm, 3.5 μm) with acetonitrile- 5 mmol/L ammonium acetate as mobile phase under gradient elution. The extract was measured directly by LC-Q TOF-MS with electrospray ionization in positive mode. The confirmatory analysis was carried out by determining the retention time and accurate masses of all compounds and fragment ions upon target MS/MS. Results The results showed that the linear range was 0.5~2000 μg/kg for each compound with the correlation coefficients of more than 0.99. The recoveries were ranged from 67.0% to 109.5%, with relative standard deviations(RSDs) less than 15.3%(n=6). Conclusion The method is simple, rapid and reliable, and is suitable for the simultaneous determination of multiresidues of veterinary drugs in aquatic products.
引文
[1]夏敏,贾丽,季怡萍.液相色谱-质谱法同时检测畜禽肉中五种大环内酯类抗生素[J].分析测试学报,2004,23:217?222.Xia M,Jia L,Ji YP.Liquid chromatography with electrospray ion-trap mass for the determination of domoic acid in shellfish[J].J Instrum Anal,2004,23:217?222.
[2]王重洋,王远鹏,王宁,等.基质固相分散-超快速液相色谱法测定牛肉中磺胺类兽药[J].分析化学,2013,41(1):83?87.Wang CY,Wang YP,Wang N,et al.Matrix solid phase dispersion coupled with ultra-fast liquid chromatography for detection of sulfonamides in beef tissue Chinese[J].J Anal Chem,2013,41(1):83?87.
[3]卜明楠,石志红,康健,等.Qu ECh ERS结合LC-MS/MS同时测定虾肉中72种兽药残留[J].分析测试学报,2012,31(5):552?558.Bu MN,Shi ZH,Kang J,et al.Simultaneous determination of 72veterinary drugs in shrimp by modified quechers and high performance liquid chromatography-tandem mass spectrometry[J].J Instrum Anal,2012,31(5):552?558.
[4]王炼,黎源倩,王海波,等.基质固相分散-超高效液相色谱-串联质谱法同时测定畜禽肉和牛奶中20种兽药残留[J].分析化学,2011,39(2):203?207.Wang L,Li YQ,Wang HB,et al.Simultaneous determination of twenty veterinary drug residues in milk and meat using matrix solid phase dispersion-ultra performance liquid chromatography-tandem mass spectrometry[J].Chinese J Anal Chem,2011,39(2):203?207.
[5]晏利芝,赵永彪,富玉,等.超高效液相色谱串联质谱法同时测定猪肉中β-受体激动剂、喹诺酮类和磺胺类兽药残留量[J].分析试验室,2011,30(1):2011?2013.Yan LZ,Zhao YB,Fu Y,et al.Simultaneous determination ofβ-agonist,quinolones and sulfonamides residues in pork by ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry[J].Chin J Anal Lab,2011,30(1):2011?2013.
[6]李娜,张玉婷,刘磊,等.Qu ECHERS超高效液相色谱串联质谱法测定动物源食品中4类29种禁限用兽药残留[J].色谱,2014,32(12):1313?1319.Li N,Zhang YT,Liu L,et al.Simultaneous determination of 4 kinds of 29banned and restricted veterinary drugs in animal-derived foods by ultra performance liquid chromatography-tandem mass spectrometry and modified Qu ECh ERS for sample preparation[J].Chin J Chromatogr,2014,32(12):1313?1319.
[7]曹亚飞,康健,常巧英,等.Qu ECHERS结合液相色谱串联质谱法快速测定奶酪中多类兽药残留[J].色谱,2015,33(2):132?139.Cao YF,Kang J,Chang QY,et al.Multi-residue determination of veterinary drugs in cheese by Qu ECh ERS and liquid chromatography–tandem mass spectrometry[J].Chin J Chromatogr,2015,33(2):132?139.
[8]赵颖,刘瑜,金雁,等.超高效液相色谱-电喷雾串联四级杆质谱法同时测定配合饲料中29种兽药[J].色谱,2012,30(9):908?914.Zhao Y,Liu Y,Jin Y,et al.Simultaneous determination of 29 veterinary drugs in compound feeds by ultra performance liquid chromatography electrospray ionization tandem quadrupole mass spectrometry[J].Chin J Chromatogr,2012,30(9):908?914.
[9]侯建波,谢文,陈笑梅,等.液相色谱-串联质谱-同位素稀释法同时测定猪肉中54种药物残留[J].质谱学报,2012,33(1):42?54.Hou JB,Xie W,Chen XM,et al.Simultaneous determination of 54 drugs residues in pork by high performance liquid chromatography-tandem mass spectrometry an isotopes dilution technique[J].J Chin Mass Spectr Soc,2012,33(1):42?54.
[10]李晓娟,彭涛,李重九.色谱-飞行时间质谱联用技术的特点及其在食品污染物分析研究中的应用[J].分析测试学报,2012,31(5):628?632.Li XJ,Peng T,Li CJ.Application progress of chromatographytime-of-flight mass spectrometry in trace analysis of food contaminants[J].J Instrum Anal,2012,31(5):628?632.
[11]刘畅,陈燕,李晓雯,等.高效液相色谱-四级杆-飞行时间质谱法同时测定猪肉中31种β-受体激动剂[J].中国食品卫生杂志,2014,26(5):464?469.Liu C,Chen Y,Li XW,et al.Determination of 31β-agonists in pork by high performance liquid chromatography-Q-time of flight mass spectrometry[J].Chin J Food Hyg,2014,26(5):464?469.
[12]彭丽,吴宁鹏,张崇威,等.超高压液相色谱-四级杆飞行时间质谱法筛查牛奶中55种药物[J].中国兽药杂志,2014,48(9):45?52.Peng L,Wu NP,Zhang CW,et al.Screening method for 55 kinds of drugs in milk by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry[J].Chin J Vet Med,2014,48(9):45?52.
[13]高馥蝶,赵妍,邵兵,等.超高效液相色谱-四级杆-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.Gao FD,Zhao Y,Shao B,et al.Determination of residues of pesticides and veterinary drugs in milk by ultra performance liquid chromatography coupled with quadrupole time of flight mass spectrometry[J].Chin J Chromatogr,2012,30(6):560?567.
[14]许成保,锁然,张峰,等.超高效液相色谱-四级杆飞行时间质谱快速检测动物饲料中10种违禁精神药物[J].色谱,2012,30(5):457?462.Xu CB,Suo R,Zhang F,et al.Simultaneous determination of 10unapproved sedative drugs in feeds by ultra-performance liquid chromatography quadrupole time-off light mass spectrometry[J].Chin J Chromatogr,2012,30(5):457?462.
[15]蓝芳,张毅,岳振峰,等.液相色谱-四级杆/飞行时间质谱快速筛查饲料中36种违禁药物[J].分析测试学报,2012,31(12):1471?1478.Lan F,Zhang Y,Yue ZF,et al.Rapid screening of 36 forbidden drugs in feeds by using liquid chromatography-quadrupole/time-of-flight mass spectrometry[J].J Instrum Anal,2012,31(12):1471?1478.
[2]王重洋,王远鹏,王宁,等.基质固相分散-超快速液相色谱法测定牛肉中磺胺类兽药[J].分析化学,2013,41(1):83?87.Wang CY,Wang YP,Wang N,et al.Matrix solid phase dispersion coupled with ultra-fast liquid chromatography for detection of sulfonamides in beef tissue Chinese[J].J Anal Chem,2013,41(1):83?87.
[3]卜明楠,石志红,康健,等.Qu ECh ERS结合LC-MS/MS同时测定虾肉中72种兽药残留[J].分析测试学报,2012,31(5):552?558.Bu MN,Shi ZH,Kang J,et al.Simultaneous determination of 72veterinary drugs in shrimp by modified quechers and high performance liquid chromatography-tandem mass spectrometry[J].J Instrum Anal,2012,31(5):552?558.
[4]王炼,黎源倩,王海波,等.基质固相分散-超高效液相色谱-串联质谱法同时测定畜禽肉和牛奶中20种兽药残留[J].分析化学,2011,39(2):203?207.Wang L,Li YQ,Wang HB,et al.Simultaneous determination of twenty veterinary drug residues in milk and meat using matrix solid phase dispersion-ultra performance liquid chromatography-tandem mass spectrometry[J].Chinese J Anal Chem,2011,39(2):203?207.
[5]晏利芝,赵永彪,富玉,等.超高效液相色谱串联质谱法同时测定猪肉中β-受体激动剂、喹诺酮类和磺胺类兽药残留量[J].分析试验室,2011,30(1):2011?2013.Yan LZ,Zhao YB,Fu Y,et al.Simultaneous determination ofβ-agonist,quinolones and sulfonamides residues in pork by ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry[J].Chin J Anal Lab,2011,30(1):2011?2013.
[6]李娜,张玉婷,刘磊,等.Qu ECHERS超高效液相色谱串联质谱法测定动物源食品中4类29种禁限用兽药残留[J].色谱,2014,32(12):1313?1319.Li N,Zhang YT,Liu L,et al.Simultaneous determination of 4 kinds of 29banned and restricted veterinary drugs in animal-derived foods by ultra performance liquid chromatography-tandem mass spectrometry and modified Qu ECh ERS for sample preparation[J].Chin J Chromatogr,2014,32(12):1313?1319.
[7]曹亚飞,康健,常巧英,等.Qu ECHERS结合液相色谱串联质谱法快速测定奶酪中多类兽药残留[J].色谱,2015,33(2):132?139.Cao YF,Kang J,Chang QY,et al.Multi-residue determination of veterinary drugs in cheese by Qu ECh ERS and liquid chromatography–tandem mass spectrometry[J].Chin J Chromatogr,2015,33(2):132?139.
[8]赵颖,刘瑜,金雁,等.超高效液相色谱-电喷雾串联四级杆质谱法同时测定配合饲料中29种兽药[J].色谱,2012,30(9):908?914.Zhao Y,Liu Y,Jin Y,et al.Simultaneous determination of 29 veterinary drugs in compound feeds by ultra performance liquid chromatography electrospray ionization tandem quadrupole mass spectrometry[J].Chin J Chromatogr,2012,30(9):908?914.
[9]侯建波,谢文,陈笑梅,等.液相色谱-串联质谱-同位素稀释法同时测定猪肉中54种药物残留[J].质谱学报,2012,33(1):42?54.Hou JB,Xie W,Chen XM,et al.Simultaneous determination of 54 drugs residues in pork by high performance liquid chromatography-tandem mass spectrometry an isotopes dilution technique[J].J Chin Mass Spectr Soc,2012,33(1):42?54.
[10]李晓娟,彭涛,李重九.色谱-飞行时间质谱联用技术的特点及其在食品污染物分析研究中的应用[J].分析测试学报,2012,31(5):628?632.Li XJ,Peng T,Li CJ.Application progress of chromatographytime-of-flight mass spectrometry in trace analysis of food contaminants[J].J Instrum Anal,2012,31(5):628?632.
[11]刘畅,陈燕,李晓雯,等.高效液相色谱-四级杆-飞行时间质谱法同时测定猪肉中31种β-受体激动剂[J].中国食品卫生杂志,2014,26(5):464?469.Liu C,Chen Y,Li XW,et al.Determination of 31β-agonists in pork by high performance liquid chromatography-Q-time of flight mass spectrometry[J].Chin J Food Hyg,2014,26(5):464?469.
[12]彭丽,吴宁鹏,张崇威,等.超高压液相色谱-四级杆飞行时间质谱法筛查牛奶中55种药物[J].中国兽药杂志,2014,48(9):45?52.Peng L,Wu NP,Zhang CW,et al.Screening method for 55 kinds of drugs in milk by ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry[J].Chin J Vet Med,2014,48(9):45?52.
[13]高馥蝶,赵妍,邵兵,等.超高效液相色谱-四级杆-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.Gao FD,Zhao Y,Shao B,et al.Determination of residues of pesticides and veterinary drugs in milk by ultra performance liquid chromatography coupled with quadrupole time of flight mass spectrometry[J].Chin J Chromatogr,2012,30(6):560?567.
[14]许成保,锁然,张峰,等.超高效液相色谱-四级杆飞行时间质谱快速检测动物饲料中10种违禁精神药物[J].色谱,2012,30(5):457?462.Xu CB,Suo R,Zhang F,et al.Simultaneous determination of 10unapproved sedative drugs in feeds by ultra-performance liquid chromatography quadrupole time-off light mass spectrometry[J].Chin J Chromatogr,2012,30(5):457?462.
[15]蓝芳,张毅,岳振峰,等.液相色谱-四级杆/飞行时间质谱快速筛查饲料中36种违禁药物[J].分析测试学报,2012,31(12):1471?1478.Lan F,Zhang Y,Yue ZF,et al.Rapid screening of 36 forbidden drugs in feeds by using liquid chromatography-quadrupole/time-of-flight mass spectrometry[J].J Instrum Anal,2012,31(12):1471?1478.