离子交换固相萃取-超高效液相色谱-串联质谱法同时测定水产品中泰妙菌素和沃尼妙林残留量
|
摘要
建立了一种强阳离子固相萃取结合超高效液相色谱-串联质谱法同时测定水产品中泰妙菌素和沃尼妙林残留量的分析方法。样品经乙腈提取和Oasis MCX固相萃取柱净化后,以乙腈-0.05%甲酸5 mmoL·L~(-1)乙酸铵水溶液为流动相,经Waters ACQUITY UPLC BEH C18色谱柱梯度洗脱分离,在电喷雾正离子多反应监测模式下,用内标法分析泰妙菌素,用外标法分析沃尼妙林。结果显示,泰妙菌素和沃尼妙林在0.1～10.0 ng·mL~(-1)内具有良好的线性关系,相关系数r均大于0.999 9,检出限和定量限分别为0.03和0.1μg·kg~(-1)。在0.1,1.0,5.0和10.0μg·kg~(-1)4种添加水平下,加标回收率在87%～114%,相对标准偏差在0.87%～6.50%。对50条鲫鱼进行实际样品验证,发现在鲫鱼的各个组织中均有泰妙菌素和沃尼妙林残留,其中肾脏残留总量最高,为453.81μg·kg~(-1),肝脏次之,为236.97μg·kg~(-1)。结果表明,所建方法能够满足对水产品中泰妙菌素和沃尼妙林残留量的分析要求。
A strong cationic solid phase extraction combined with ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was developed for the simultaneous determination of tiamulin and valnemulin residues in aquatic products. The sample was extracted with acetonitrile and purified with Oasis MCX SPE cartridge,then was eluted with a gradient of acetonitrile-0.05 % formic acid in 5 mmoL·L~(-1) ammonium acetate and eluted with a Waters ACQUITY UPLC BEH C18 column. In the electrospray positive ionization multiple reaction monitoring mode, tiamulin was analyzed by internal standard method and valnemulin was analyzed by external standard method.The results showed that there was a good linear relationship between tiamulin and valnemulin in the range of 0.1～10.0 ng·mL~(-1) with the correlation coefficients r is greater than 0.999 9. The limits of detection and quantification limits were 0.03 and 0.1 μg·kg~(-1), respectively. The spiked recoveries were between 87 % and 114 % with the relative standard deviations between 0.87 % and 6.50 % at the four addition levels of 0.1, 1.0, 5.0 and 10.0 μg·kg~(-1).Through the actual sample verification of 50 carp, there were residues in the tissues of the carp with the highest content being 453.81 μg·kg~(-1) in kidney and the second content being 236.97 μg·kg~(-1) in liver. The results show that the established method can meet the analytical requirements of tiamulin and valnemulin residues in aquatic products.
A strong cationic solid phase extraction combined with ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was developed for the simultaneous determination of tiamulin and valnemulin residues in aquatic products. The sample was extracted with acetonitrile and purified with Oasis MCX SPE cartridge,then was eluted with a gradient of acetonitrile-0.05 % formic acid in 5 mmoL·L~(-1) ammonium acetate and eluted with a Waters ACQUITY UPLC BEH C18 column. In the electrospray positive ionization multiple reaction monitoring mode, tiamulin was analyzed by internal standard method and valnemulin was analyzed by external standard method.The results showed that there was a good linear relationship between tiamulin and valnemulin in the range of 0.1～10.0 ng·mL~(-1) with the correlation coefficients r is greater than 0.999 9. The limits of detection and quantification limits were 0.03 and 0.1 μg·kg~(-1), respectively. The spiked recoveries were between 87 % and 114 % with the relative standard deviations between 0.87 % and 6.50 % at the four addition levels of 0.1, 1.0, 5.0 and 10.0 μg·kg~(-1).Through the actual sample verification of 50 carp, there were residues in the tissues of the carp with the highest content being 453.81 μg·kg~(-1) in kidney and the second content being 236.97 μg·kg~(-1) in liver. The results show that the established method can meet the analytical requirements of tiamulin and valnemulin residues in aquatic products.
引文
[1] POULSEN S M, KARLSSON M, JOHANSSON L B, et al. The pleuromutilin drugs tiamulin and valnemulin bind to the RNA at the peptidyl transferase centre on the ribosome[J].Molecular Microbiology,2001, 41(5):1091.
[2] XIAO X, SUN J, YANG T, et al. In vivo pharmacokinetic/pharmacodynamic profiles of valnemulin in an experimental intratracheal mycoplasma gallisepticum infection model[J].Antimicrobial Agents&Chemotherapy, 2015, 59(7):3754.
[3]赵东豪,朱理想,贺利民,等.沃尼妙林在电喷雾质谱中的行为研究[J].质谱学报, 2009, 30(05):311-313.ZHAO D H, ZHU L X, HE L M, et al.Study on the behavior of valnemulin in electrospray ionization mass spectrometry[J].Journal of Chinese Mass Spectrometry Society, 2009,30(5):311-313.
[4] JACOBS M R. Retapamulin:A semisynthetic pleuromutilin compound for topical treatment of skin infections in adults and children[J].Future Microbiology, 2007,2(6):591-600.
[5] B?SLING J, POULSEN S M, VESTER B, et al.Resistance to the peptidyl transferase inhibitor tiamulin caused by mutation of ribosomal proteinl3[J].Antimicrobial Agents&Chemotherapy, 2003, 47(9):2892-2896.
[6] POULSEN S M, KARLSSON M, JOHANSSON L B, et al. The pleuromutilin drugs tiamulin and valnemulin bind to the RNA at the peptidyl transferase centre on the ribosome[J].Molecular Microbiology,2001, 41(5):1091.
[7] BAYNES R E, DEDONDER K, KISSELL L, et al.Health concerns and management of select veterinary drug residues[J].Food&Chemical Toxicology:An International Journal Published for the British Industrial Biological Research Association, 2016,88:112-122.
[8] CERNIGLIA C E, PINEIRO S A, KOTARSKI S F. An update discussion on the current assessment of the safety of veterinary antimicrobial drug residues in food with regard to their impact on the human intestinal microbiome[J].Drug Testing&Analysis,2016, 8(5/6):539-548.
[9] ZHANG Y Y, XU K P, REN D, et al. Synthesis and antibacterial activities of pleuromutilin derivatives[J].Chinese Chemical Letters, 2009, 20(1):29-31.
[10] JONES R N, PFALLER M A, RHOMBERG P R,et al. Tiamulin activity against fastidious and nonfastidious veterinary and human bacterial isolates:Initial development of in vitro susceptibility test methods[J].Journal of Clinical Microbiology, 2002,40(2):461-465.
[11] HUI L, WANG Y, LI X, et al. Determination of valnemulin in swine and bovine tissues by ultra-high performance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography B Analytical Technologies in the Biomedical&Life Sciences, 2016, 1014:102-106.
[12]陈兴荣,刘晓莎,方炳虎,等.高效液相色谱法测定沃尼妙林含量的研究[J].黑龙江畜牧兽医,2011(3):120-122.CHEN X R, LIU X S, FANG B H, et al.Determination of valnemulin content by high performance liquid chromatography[J].Heilongjiang Animal Science and Veterinary Medicine, 2011(3):120-122.
[13]程林丽,张素霞,沈建忠,等.高效液相色谱法快速检测猪组织中泰妙菌素残留[J].分析化学,2009,37(5):718-720.CHENG L L, ZHANG S X, SHEN J Z, et al.Rapid detection of tiamulin residues in porcine tissues by high performance liquid chromatography[J].Chinese Journal of Analytical Chemistry, 2009, 37(5):718-720.
[14] CHEN H C, CHENG S H, TSAI Y H, et al.Determination of tiamulin residue in pork and chicken by solid phase extraction and HPLC[J].Journal of Food and Drug Analysis, 2006, 14(1):80-83.
[15] MOORE D B, BRITTON N L, SMALLIDGE R L, et al. Determination of tiamulin in type C medicated swine feeds using high throughput extraction with liquid chromatography[J].Journal of Aoac International, 2002, 85(3):533-540.
[16] BAERE S D, DEVREESE M, AN M, et al.Quantification of 8-α-hydroxy mutilin as marker residue for tiamulin in rabbit tissues by highperformance liquid chromatography-mass spectrometry[J].Analytical&Bioanalytical Chemistry, 2015, 407(15):4437-4445.
[17]颜虎.沃尼妙林在肉鸡体内的药动学及其组织残留研究[D].南昌:江西农业大学,2012.YAN H.Study on Pharmacokinetics and Tissue Residues of Valnemulin in Broilers[D]. Nanchang:Jiangxi Agricultural University,2012.
[18] XIA X, JIAN S, TAO Y, et al. Pharmacokinetic/Pharmacodynamic profiles of tiamulin in an experimental intratracheal infection model of mycoplasma gallisepticum[J].Frontiers in Veterinary Science, 2016, 3(5):75.
[19] SCHLüSENER M P, BESTER K, SPITELLER M. Determination of antibiotics such as macrolides,ionophores and tiamulin in liquid manure by HPLCMS/MS[J].Analytical&Bioanalytical Chemistry,2003, 375(7):942-947.
[20] NOZAL M J, BERNAL J L, MARTíN M T, et al.Trace analysis of tiamulin in honey by liquid chromatography-diode array-electrospray ionization mass spectrometry detection[J].Journal of Chromatography A, 2006, 1116(1/2):102-108.
[21] YIN Z, CHAI T, MU P, et al. Multi-residue determination of 210 drugs in pork by ultra-highperformance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2016, 1463:49-59.
[22] LI H, WANG Y, LI X, et al. Determination of valnemulin in swine and bovine tissues by ultra-high performance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography B,2016, 1014:102-106.
[23]隋涛,付建,李晓玉,等.超高效液相色谱串联质谱法同时测定水产品中6种兽药残留[J].食品科学,2011,32(10):203-207.SUI T, FU J, LI X Y, et al. Simultaneous determination of veterinary drug residues in aquatic products by UPLC-MS/MS[J].Food Science, 2011,32(10):203-207.
[24]叶妮,尹晖,王亦琳,等.动物性食品中泰妙菌素残留标志物检测UPLC-MS/MS法研究[J].中国兽药杂志,2016,50(11):49-53.YE N,YIN H,WANG Y L, et al. The research of 8-α-hydroxy mutilin residues in animal derived food by UPLC-MS/MS[J].Chinese Journal of Veterinary Drug,2016,50(11):49-53.
[25]王蕊,远立国,贺利民,等.液相色谱-串联质谱法测定鸡组织中沃尼妙林残留[J].分析化学,2010,38(07):1031-1035.WANG R, YUAN L G, HE L M, et al.Determination of valnemulin residues in chicken tissues by liquid chromatography-tandem mass spectrometry[J].Chinese Journal of Analytical Chemistry,2010,38(07):1031-1035.
[26]黄雅丽.固相萃取-高效液相色谱-串联质谱法测定禽畜和水产品中沃尼妙林和泰妙菌素[J].分析试验室,2010,29(1):103-106.HUANG Y L. Solid phase extraction-high performance liquid chromatography-tandemmass spectrometry for the determination of valnemullin and tiamulin in livestock and aquatic products[J].Chinese Journal of Analysis Laboratory,2010,29(1):103-106.
[27]耿士伟,朱志谦,曲斌,等.高效液相-串联质谱法测定鸡肉中泰妙菌素残留的研究[J].中国家禽,2012,34(11):28-31.GENG S W, ZHU Z Q, QU B, et al.Determination of Tiamulin residues in chicken meat by HPLC-MS/MS[J].China Poultry,2012,34(11):28-31.
[28]国家质量监督检验检疫局.出口活鱼泰妙菌素检测技术规范:SN/T 4483-2016[S].北京:中国标准出版社, 2016.General Administration of Quality Supervision,Inspection and Quarantine of the People’s Republic of China.Export of Live Fish Tiamulin Test Specification:SN/T 4483-2016[S]. Beijing:China Standard Press, 2016.
[2] XIAO X, SUN J, YANG T, et al. In vivo pharmacokinetic/pharmacodynamic profiles of valnemulin in an experimental intratracheal mycoplasma gallisepticum infection model[J].Antimicrobial Agents&Chemotherapy, 2015, 59(7):3754.
[3]赵东豪,朱理想,贺利民,等.沃尼妙林在电喷雾质谱中的行为研究[J].质谱学报, 2009, 30(05):311-313.ZHAO D H, ZHU L X, HE L M, et al.Study on the behavior of valnemulin in electrospray ionization mass spectrometry[J].Journal of Chinese Mass Spectrometry Society, 2009,30(5):311-313.
[4] JACOBS M R. Retapamulin:A semisynthetic pleuromutilin compound for topical treatment of skin infections in adults and children[J].Future Microbiology, 2007,2(6):591-600.
[5] B?SLING J, POULSEN S M, VESTER B, et al.Resistance to the peptidyl transferase inhibitor tiamulin caused by mutation of ribosomal proteinl3[J].Antimicrobial Agents&Chemotherapy, 2003, 47(9):2892-2896.
[6] POULSEN S M, KARLSSON M, JOHANSSON L B, et al. The pleuromutilin drugs tiamulin and valnemulin bind to the RNA at the peptidyl transferase centre on the ribosome[J].Molecular Microbiology,2001, 41(5):1091.
[7] BAYNES R E, DEDONDER K, KISSELL L, et al.Health concerns and management of select veterinary drug residues[J].Food&Chemical Toxicology:An International Journal Published for the British Industrial Biological Research Association, 2016,88:112-122.
[8] CERNIGLIA C E, PINEIRO S A, KOTARSKI S F. An update discussion on the current assessment of the safety of veterinary antimicrobial drug residues in food with regard to their impact on the human intestinal microbiome[J].Drug Testing&Analysis,2016, 8(5/6):539-548.
[9] ZHANG Y Y, XU K P, REN D, et al. Synthesis and antibacterial activities of pleuromutilin derivatives[J].Chinese Chemical Letters, 2009, 20(1):29-31.
[10] JONES R N, PFALLER M A, RHOMBERG P R,et al. Tiamulin activity against fastidious and nonfastidious veterinary and human bacterial isolates:Initial development of in vitro susceptibility test methods[J].Journal of Clinical Microbiology, 2002,40(2):461-465.
[11] HUI L, WANG Y, LI X, et al. Determination of valnemulin in swine and bovine tissues by ultra-high performance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography B Analytical Technologies in the Biomedical&Life Sciences, 2016, 1014:102-106.
[12]陈兴荣,刘晓莎,方炳虎,等.高效液相色谱法测定沃尼妙林含量的研究[J].黑龙江畜牧兽医,2011(3):120-122.CHEN X R, LIU X S, FANG B H, et al.Determination of valnemulin content by high performance liquid chromatography[J].Heilongjiang Animal Science and Veterinary Medicine, 2011(3):120-122.
[13]程林丽,张素霞,沈建忠,等.高效液相色谱法快速检测猪组织中泰妙菌素残留[J].分析化学,2009,37(5):718-720.CHENG L L, ZHANG S X, SHEN J Z, et al.Rapid detection of tiamulin residues in porcine tissues by high performance liquid chromatography[J].Chinese Journal of Analytical Chemistry, 2009, 37(5):718-720.
[14] CHEN H C, CHENG S H, TSAI Y H, et al.Determination of tiamulin residue in pork and chicken by solid phase extraction and HPLC[J].Journal of Food and Drug Analysis, 2006, 14(1):80-83.
[15] MOORE D B, BRITTON N L, SMALLIDGE R L, et al. Determination of tiamulin in type C medicated swine feeds using high throughput extraction with liquid chromatography[J].Journal of Aoac International, 2002, 85(3):533-540.
[16] BAERE S D, DEVREESE M, AN M, et al.Quantification of 8-α-hydroxy mutilin as marker residue for tiamulin in rabbit tissues by highperformance liquid chromatography-mass spectrometry[J].Analytical&Bioanalytical Chemistry, 2015, 407(15):4437-4445.
[17]颜虎.沃尼妙林在肉鸡体内的药动学及其组织残留研究[D].南昌:江西农业大学,2012.YAN H.Study on Pharmacokinetics and Tissue Residues of Valnemulin in Broilers[D]. Nanchang:Jiangxi Agricultural University,2012.
[18] XIA X, JIAN S, TAO Y, et al. Pharmacokinetic/Pharmacodynamic profiles of tiamulin in an experimental intratracheal infection model of mycoplasma gallisepticum[J].Frontiers in Veterinary Science, 2016, 3(5):75.
[19] SCHLüSENER M P, BESTER K, SPITELLER M. Determination of antibiotics such as macrolides,ionophores and tiamulin in liquid manure by HPLCMS/MS[J].Analytical&Bioanalytical Chemistry,2003, 375(7):942-947.
[20] NOZAL M J, BERNAL J L, MARTíN M T, et al.Trace analysis of tiamulin in honey by liquid chromatography-diode array-electrospray ionization mass spectrometry detection[J].Journal of Chromatography A, 2006, 1116(1/2):102-108.
[21] YIN Z, CHAI T, MU P, et al. Multi-residue determination of 210 drugs in pork by ultra-highperformance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2016, 1463:49-59.
[22] LI H, WANG Y, LI X, et al. Determination of valnemulin in swine and bovine tissues by ultra-high performance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography B,2016, 1014:102-106.
[23]隋涛,付建,李晓玉,等.超高效液相色谱串联质谱法同时测定水产品中6种兽药残留[J].食品科学,2011,32(10):203-207.SUI T, FU J, LI X Y, et al. Simultaneous determination of veterinary drug residues in aquatic products by UPLC-MS/MS[J].Food Science, 2011,32(10):203-207.
[24]叶妮,尹晖,王亦琳,等.动物性食品中泰妙菌素残留标志物检测UPLC-MS/MS法研究[J].中国兽药杂志,2016,50(11):49-53.YE N,YIN H,WANG Y L, et al. The research of 8-α-hydroxy mutilin residues in animal derived food by UPLC-MS/MS[J].Chinese Journal of Veterinary Drug,2016,50(11):49-53.
[25]王蕊,远立国,贺利民,等.液相色谱-串联质谱法测定鸡组织中沃尼妙林残留[J].分析化学,2010,38(07):1031-1035.WANG R, YUAN L G, HE L M, et al.Determination of valnemulin residues in chicken tissues by liquid chromatography-tandem mass spectrometry[J].Chinese Journal of Analytical Chemistry,2010,38(07):1031-1035.
[26]黄雅丽.固相萃取-高效液相色谱-串联质谱法测定禽畜和水产品中沃尼妙林和泰妙菌素[J].分析试验室,2010,29(1):103-106.HUANG Y L. Solid phase extraction-high performance liquid chromatography-tandemmass spectrometry for the determination of valnemullin and tiamulin in livestock and aquatic products[J].Chinese Journal of Analysis Laboratory,2010,29(1):103-106.
[27]耿士伟,朱志谦,曲斌,等.高效液相-串联质谱法测定鸡肉中泰妙菌素残留的研究[J].中国家禽,2012,34(11):28-31.GENG S W, ZHU Z Q, QU B, et al.Determination of Tiamulin residues in chicken meat by HPLC-MS/MS[J].China Poultry,2012,34(11):28-31.
[28]国家质量监督检验检疫局.出口活鱼泰妙菌素检测技术规范:SN/T 4483-2016[S].北京:中国标准出版社, 2016.General Administration of Quality Supervision,Inspection and Quarantine of the People’s Republic of China.Export of Live Fish Tiamulin Test Specification:SN/T 4483-2016[S]. Beijing:China Standard Press, 2016.