超高效液相色谱-串联质谱法测定多脂肪类动物源性食品中硝基呋喃代谢物
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摘要
目的建立多脂肪类动物源性样品中4种硝基呋喃代谢物的超高效液相色谱-串联质谱(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)的快速检测方法。方法样品利用硅藻土充分研磨后,增加0.125 mol/L盐酸用量至25 mL进行衍生,调节pH值后,使用32 mL提取溶剂乙酸乙酯,分2次提取,提取溶剂合并旋干后,加入定容试剂,超声提取1.5 min,加入2.5 mL正己烷提取脂肪,水相过滤上机检测。分析物采用电喷雾电离,正离子扫描,多反应检测模式(multiple reaction monitoring,MRM),基质匹配内标标准曲线法定量。结果 4种代谢物的工作曲线在0.5~30.0μg/L浓度范围内线性良好,相关系数r~2均达到了0.998以上,5-吗啉甲基-3-氨基-2-恶唑烷基酮(3-amino-5-morpholinomethyl-2-oxazolidinone,AMOZ)、1-氨基-乙内酰脲(1-amino-hydantoin,AHD)、3-氨基-2-恶唑酮(3-amino-2-oxalidinone,AOZ)、氨基脲(semicarbazide,SEM)最低检出限分别为0.052、0.043、0.049、0.108μg/kg。平均回收率在84.5%~105.4%之间,相对标准偏差1.26%~7.58%。结论方法解决了多脂类动物源性样品中硝基呋喃代谢物易干扰、回收低、检不准的难点,同时使用4种硝基呋喃代谢物内标定量结果,结果准确可靠、适用多脂肪类样品中硝基呋喃代谢物的检测。
Objective To establish a analytical method for detection of metabolites of nitrofuran in fatty animal samples by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Methods The derivatives of samples were generated in 25 mL hydrochloric acid(0.125 mol/L) and extracted by 32 mL ethyl acetate after being grinded with diatomaceous earth. Before extraction of fat, nitrofuran metabolites in pear-shaped bottle were extracted by ultrasonic about 1.5 min. Identification was achieved by electrospray ionization in positive mode(ESI+) using multiple reaction monitoring(MRM). Results The calibration curves of the 4 analytes showed a good linearity in the concentrations of 0.5-30 μg/L with r~2 above 0.998. The limits of detection of 3-amino-5-morpholinomethyl-2-oxazolidinone(AMOZ), 1-amino-hydantoin(AHD), 3-amino-2-oxalidinone(AOZ),and semicarbazide(SEM) were 0.052, 0.043, 0.049, and 0.108 μg/kg, respectively. The average recoveries were 84.5%-105.4% with relative standard deviations(RSDs) of 1.26%-7.58%. Conclusion This method solves the difficulties of easy interference, low recovery and inaccuracy of fatty animal samples of nitrofuran metabolites, and is accurate and reliable with internal standard, which can be applied for detection of fatty animal samples of nitrofuran metabolites.
Objective To establish a analytical method for detection of metabolites of nitrofuran in fatty animal samples by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Methods The derivatives of samples were generated in 25 mL hydrochloric acid(0.125 mol/L) and extracted by 32 mL ethyl acetate after being grinded with diatomaceous earth. Before extraction of fat, nitrofuran metabolites in pear-shaped bottle were extracted by ultrasonic about 1.5 min. Identification was achieved by electrospray ionization in positive mode(ESI+) using multiple reaction monitoring(MRM). Results The calibration curves of the 4 analytes showed a good linearity in the concentrations of 0.5-30 μg/L with r~2 above 0.998. The limits of detection of 3-amino-5-morpholinomethyl-2-oxazolidinone(AMOZ), 1-amino-hydantoin(AHD), 3-amino-2-oxalidinone(AOZ),and semicarbazide(SEM) were 0.052, 0.043, 0.049, and 0.108 μg/kg, respectively. The average recoveries were 84.5%-105.4% with relative standard deviations(RSDs) of 1.26%-7.58%. Conclusion This method solves the difficulties of easy interference, low recovery and inaccuracy of fatty animal samples of nitrofuran metabolites, and is accurate and reliable with internal standard, which can be applied for detection of fatty animal samples of nitrofuran metabolites.
引文
[1]陈威风,陈敬鑫.肉制品中硝基呋喃类药物残留检测方法的研究进展[J].肉类研究,2011,25(12):53-57.Chen WF,Chen JX.Recent advances in research on nitrofuran residues in meat products[J].Meat Res,2011,25(12):53-57.
[2]王媛,蔡友琼,贾东芬,等.液相色谱串联质谱法测定水产品中硝基呋喃类代谢物残留量[J].分析实验室,2009,28(12):86-89.Wang Y,Cai YQ,Jia DF,et al.Determination of nitrofuran metabolite residues in aquatic products by high performance liquid chromatography[J].Chin J Anal Lab,2009,28(12):86-89.
[3]Zuidema T,Mulder PJ,Rhijn JA,et al.ELISA for semicarbazide and its application for screening in food contamination[J].Anal Chin Acta,2005,529:339.
[4]Aila O,Shitandi A,Mahungu MS,et al.Determination of nitrofuran drugs in aquatic products[J].Food Control,2009,20:543.
[5]Council Regulation.Residual levels of malachite green and nitrofuran metabolites in farmed fish[J].Off J Eur Commun,1990,224:1.
[6]田蕴,罗晓琴.水产品硝基呋喃类药物残留检测[J].动物保健,2006,7:45-47.Tian Y,Luo XQ.Determination of nitrofuran drugs in aquatic products[J].Anim Health,2006,7:45-47.
[7]赵善仓,李增梅,刘宾,等.超高效液相色谱串联质谱法测定畜产品中硝基呋喃类抗生素代谢物残留的研究[J].中国兽药杂志,2008,42(7):17-21.Zhao SC,Li ZM,Liu B,et al.Determination of the metabolites of nitrofuran antibiotics residues in animal tissues by UPLC-MS/MS[J].Chin J Vet Drug,2008,42(7):17-21.
[8]林黎明,林回春,刘心同,等.固相萃取高效液相色谱-质谱法测定动物组织中硝基呋喃代谢产物[J].分析化学,2005,33(5):707-7l0.Lin LM,Lin HC,Liu XT,et al.Determination of nitrofuran metablolites in animal tissues by high performance liquid chromatography-mass spectrometry and study of their residual discipline[J].Chin J Anal Chem,2005,33(5):707-710.
[9]Xia X,Li X,Zhang S,et al.Research progress of the origin of semicarbazide in food[J].Chromatogr A,2008,1208:101.
[10]Mcneil EM,Ritchie AM,Melton DW.The toxicity of nitrofuran compounds on melanoma and neuroblastoma cells is enhanced by olaparib and ameliorated by melanin pigment[J].DNA Repair,2013,12(11):1000-1006.
[11]魏法山,盖圣美,谢文佳,等.动物源性食品中硝基呋喃类兽药残留检测方法的研究进展[J].食品安全质量检测学报,2016,7(6):2289-2295Wei FS,Gai SM,Xie WJ,et al.Progress on determination of nitrofuran and their metabolites in animal-derived food[J].J Food Saf Qual,2016,7(6):2289-2295
[12]Chumanee S,Sutthivaiyakit S,Sutthivaiyakit P.New reagent for trace determination of protein-bound metabolites of nitrofurans in shrimp using liquid chromatography with diode array detector[J].J Agric Food Chem,2009,57:1752-1759.
[13]GB/T 21311-2007动物源性食品中硝基呋喃类药物代谢物残留量检测方法-高效液相色谱/串联质谱法[S].GB/T 21311-2007 Determination of residues of nitrofuran metabolites in foodstuffs of animal origin-HPLC-MS/MS[S].
[14]司红彬,梁军.硝基呋喃类药物的残留检测(下)[J].兽医导刊,2007,10(122):40-42.Si HB,Liang J.Determination of residues of nitrofuran metabolites[J].Vet Guid,2007,10(122):40-42.
[15]王习达,陈辉,左建忠,等.水产品中硝基呋喃类药物残留的检测与控制[J].现代农业科技,2007,18:152-155.Wang XD,Chen H,Zuo JZ.Determination and control of nitrofuran metabolite residues in aquatic products[J].Mod Agric Technol,2007,18:152-155.
[2]王媛,蔡友琼,贾东芬,等.液相色谱串联质谱法测定水产品中硝基呋喃类代谢物残留量[J].分析实验室,2009,28(12):86-89.Wang Y,Cai YQ,Jia DF,et al.Determination of nitrofuran metabolite residues in aquatic products by high performance liquid chromatography[J].Chin J Anal Lab,2009,28(12):86-89.
[3]Zuidema T,Mulder PJ,Rhijn JA,et al.ELISA for semicarbazide and its application for screening in food contamination[J].Anal Chin Acta,2005,529:339.
[4]Aila O,Shitandi A,Mahungu MS,et al.Determination of nitrofuran drugs in aquatic products[J].Food Control,2009,20:543.
[5]Council Regulation.Residual levels of malachite green and nitrofuran metabolites in farmed fish[J].Off J Eur Commun,1990,224:1.
[6]田蕴,罗晓琴.水产品硝基呋喃类药物残留检测[J].动物保健,2006,7:45-47.Tian Y,Luo XQ.Determination of nitrofuran drugs in aquatic products[J].Anim Health,2006,7:45-47.
[7]赵善仓,李增梅,刘宾,等.超高效液相色谱串联质谱法测定畜产品中硝基呋喃类抗生素代谢物残留的研究[J].中国兽药杂志,2008,42(7):17-21.Zhao SC,Li ZM,Liu B,et al.Determination of the metabolites of nitrofuran antibiotics residues in animal tissues by UPLC-MS/MS[J].Chin J Vet Drug,2008,42(7):17-21.
[8]林黎明,林回春,刘心同,等.固相萃取高效液相色谱-质谱法测定动物组织中硝基呋喃代谢产物[J].分析化学,2005,33(5):707-7l0.Lin LM,Lin HC,Liu XT,et al.Determination of nitrofuran metablolites in animal tissues by high performance liquid chromatography-mass spectrometry and study of their residual discipline[J].Chin J Anal Chem,2005,33(5):707-710.
[9]Xia X,Li X,Zhang S,et al.Research progress of the origin of semicarbazide in food[J].Chromatogr A,2008,1208:101.
[10]Mcneil EM,Ritchie AM,Melton DW.The toxicity of nitrofuran compounds on melanoma and neuroblastoma cells is enhanced by olaparib and ameliorated by melanin pigment[J].DNA Repair,2013,12(11):1000-1006.
[11]魏法山,盖圣美,谢文佳,等.动物源性食品中硝基呋喃类兽药残留检测方法的研究进展[J].食品安全质量检测学报,2016,7(6):2289-2295Wei FS,Gai SM,Xie WJ,et al.Progress on determination of nitrofuran and their metabolites in animal-derived food[J].J Food Saf Qual,2016,7(6):2289-2295
[12]Chumanee S,Sutthivaiyakit S,Sutthivaiyakit P.New reagent for trace determination of protein-bound metabolites of nitrofurans in shrimp using liquid chromatography with diode array detector[J].J Agric Food Chem,2009,57:1752-1759.
[13]GB/T 21311-2007动物源性食品中硝基呋喃类药物代谢物残留量检测方法-高效液相色谱/串联质谱法[S].GB/T 21311-2007 Determination of residues of nitrofuran metabolites in foodstuffs of animal origin-HPLC-MS/MS[S].
[14]司红彬,梁军.硝基呋喃类药物的残留检测(下)[J].兽医导刊,2007,10(122):40-42.Si HB,Liang J.Determination of residues of nitrofuran metabolites[J].Vet Guid,2007,10(122):40-42.
[15]王习达,陈辉,左建忠,等.水产品中硝基呋喃类药物残留的检测与控制[J].现代农业科技,2007,18:152-155.Wang XD,Chen H,Zuo JZ.Determination and control of nitrofuran metabolite residues in aquatic products[J].Mod Agric Technol,2007,18:152-155.