液相色谱-串联质谱法测定贝类组织中硝基呋喃类代谢物残留量
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摘要
建立了液相色谱-串联质谱法测定贝类产品中硝基呋喃类代谢物残留量的分析方法.贝类组织中残留的硝基呋喃类蛋白结合态代谢物在酸性条件下水解,用2-硝基苯甲醛衍生化,经乙酸乙酯液-液萃取、浓缩及基质分散固相萃取和超滤净化后,采用液相色谱串联质谱仪测定,以电喷雾离子源正离子扫描模式进行质谱分析,内标法定量.硝基呋喃类代谢物在0.5—20μg·L~(-1)范围内线性关系良好,相关系数R2>0.999.4种硝基呋喃类代谢物添加水平为1.00,2.50、10.0μg·kg~(-1)时,平均回收率在90.1%—99.5%之间,批内和批间变异系数均<15%.4种硝基呋喃类代谢物的检出限均为0.50μg·kg~(-1).本方法灵敏、高效、简单、重现性好,满足贝类产品中硝基呋喃类代谢物残留量的检测要求.
A high performance liquid chromatography-tandem mass spectrometry( HPLC-MS/MS)method was developed for the determination of nitrofuran metabolites residues in shellfish tissues. The residues of protein adducts from nitrofuran metabolites in shellfish tissues were hydrolyzed by HCl,followed by derivatizing with nitrobenzaldehyde. The resulting derivatives were extracted with ethyl acetate,and the organic layer was concentrated by a stream of nitrogen gas. After purification by matrix dispersed solid phase extraction combined with ultrafiltration,the samples were analyzed by LC-MS/MS using internal standard method. Electrospray ionization was applied and operated in positive ion mode. The calibration curve showed a good linearity in the range of 0.5—20 μg·L~(-1) with the correlation coefficient over 0.999. The average recoveries for the spiked samples at concentrations of 1.00,2.50 and 10. 0 μg·kg~(-1) ranged from 90. 1% to 99. 5% with intra and inter batch relative standard deviations below 15%. The limit of detection of nitrofuran metabolites was 0.50 μg·kg~(-1).This approach is proved to be a simple and efficient method to identify and quantify nitrofuran metabolites in shellfish tissues with satisfactory sensitivity and repeatability.
A high performance liquid chromatography-tandem mass spectrometry( HPLC-MS/MS)method was developed for the determination of nitrofuran metabolites residues in shellfish tissues. The residues of protein adducts from nitrofuran metabolites in shellfish tissues were hydrolyzed by HCl,followed by derivatizing with nitrobenzaldehyde. The resulting derivatives were extracted with ethyl acetate,and the organic layer was concentrated by a stream of nitrogen gas. After purification by matrix dispersed solid phase extraction combined with ultrafiltration,the samples were analyzed by LC-MS/MS using internal standard method. Electrospray ionization was applied and operated in positive ion mode. The calibration curve showed a good linearity in the range of 0.5—20 μg·L~(-1) with the correlation coefficient over 0.999. The average recoveries for the spiked samples at concentrations of 1.00,2.50 and 10. 0 μg·kg~(-1) ranged from 90. 1% to 99. 5% with intra and inter batch relative standard deviations below 15%. The limit of detection of nitrofuran metabolites was 0.50 μg·kg~(-1).This approach is proved to be a simple and efficient method to identify and quantify nitrofuran metabolites in shellfish tissues with satisfactory sensitivity and repeatability.
引文
[1]李芳,陈莹,李献刚,等.动物源性食品中硝基呋喃及其代谢物产物检测方法研究进展[J].食品安全质量检测学报,2016,7(6):2321-2327.LI F,CHEN Y,LI X G,et al. Progress on determination of nitrofuran and their metabolites in animal-derived food[J]. Journal of FoodSafety and Quality,2016,7(6):2321-2327(in Chinese).
[2] TAO Y F,YU H,CHEN D M,et al. Determination of sodium nifurstyrenate and nitrovin residues in edible food by liquid chromatography-tandem mass spectrometry after ultrasound-assisted extraction[J]. Journal of Chromatography B,2010,878:3415-3420.
[3]邢丽红,李兆新,孙伟红,等.液相色谱-串联质谱法检测水产品中硝基呋喃类药物的残留量[J].食品安全质量检测学报,2017,8(4):1233-1239.XING L H,LI Z X,SUN W H,et al. Simultaneous determination of four kinds of nitrofurans residues in aquatic products by liquidchromatography-tandem mass spectrometry[J]. Journal of Food Safety and Quality,2017,8(4):1233-1239(in Chinese).
[4] KAUFMANN A,BUTCHER P,MADEN K,et al. Determination of nitrofuran and chloramphenicol residues by high resolution massspectrometry versus tandem quadrupole mass spectrometry[J]. Analytica Chimica Acta,2015,862:41-52.
[5] VERDON E,COUEDOR P,SANDERS P. Multi-residue monitoring for the simultaneous determination of five nitrofurans(furazolidone,furaltadone,nitrofurazone,nitrofurantoine,nifursol)in poultry muscle tissue through the detection of their five major metabolites(AOZ,AMOZ,SEM,AHD,DNSAH)by liquid chromatography coupled to electrospray tandem mass spectrometry in house validation in line withcommission decision 657/2002/EC[J]. Analytica Chimica Acta,2007,586:336-347.
[6] KAUFAMANN A,BUTCHER P,MADEN K. LC-MS/MS method for determing nifursol and other nitrofuran residues in meat[J]. MittLebensm Hyg,2004,95:135-146.
[7]唐红梅,曾芳,李成洪.食品中硝基呋喃类药物及其代谢物残留检测的研究进展[J].食品安全质量检测学报,2016,7(10):3952-3954.TANG H M,ZENG F,LI C H. Progress on the detection of nitrofurans drugs residues and their metabolites in food[J]. Journal of FoodSafety and Quality,2016,7(10):3952-3954(in Chinese).
[8] FERNANDO R,MUNASINGHE D M S,GUNASENA A R C,et al. Determination of nitrofuran metabolites in shrimp muscle by liquidchromatography-photo diode array detection[J]. Food Control,2017,72:300-305.
[9] DU N N,CHEN M M,SHENG L Q,et al. Determination of nitrofuran metabolites in shrimp by high performance liquid chromatographywith fluorescence detection and liquid chromatography-tandem mass spectrometry using a new derivatization reagent[J]. Journal ofChromatography A,2014,1327:90-96.
[10] ZHANG Y B,QIAO H O,CHEN C,et al. Determination of nitrofurans metabolites residues in aquatic products by ultra-performance liquidchromatography-tandem mass spectrometry[J]. Food Chemistry,2016,192:612-617.
[11] ZHAI H Y,ZHANG L C,PAN Y F,et al. Simultaneous determination of chloramphenicol,ciprofloxacin,nitrofuran antibiotics and theirmetabolites in fishery products by CE[J]. Chromatographia,2015,78:551-556.
[12] JIANG W X,LUO P J,WANG X. Development of an enzyme-linked immunosorbent assay for the detection of nitrofurantoin metabolite,1-amino-hydantoin,in animal tissues[J]. Food Control,2012,23:20-25.
[13] ZHANG Z W,WU Y P,LI X W,et al. Multi-class method for the determination of nitroimidazoles,nitrofurans,and chloramphenicol inchicken muscle and egg by dispersive-solid phase extraction and ultra-high performance liquid chromatography-tandem mass spectrometry[J]. Food Chemistry,2017,217:182-190.
[14] DONGKYUM K,BYUNGJOO K,SEOK-WON H. An optimized method for the accurate determination of nitrofurans in chicken meat usingisotope dilution-liquid chromatography/mass spectrometry[J]. Journal of Food Composition and Analysis,2015,40:24-31.
[15]孙稚菁,王婵,李崇,等.高效液相色谱-串联质谱法快速检测海参中硝基呋喃代谢物前处理方案的探讨[J].分析测试学报,2016,35(9):1127-1131.SUN Z J,WANG C,LI C,et al. Pretreatment programs discussion of detecting nitrofuran metabolites rapidly in holothurians by highperformance liquid chromatography tandem mass spectrometry[J]. Journal of Instrumental Analysis,2016,35(9):1127-1131(inChinese).
[16]万建春,王文君,占春瑞,等.高效液相色谱-串联质谱法同时测定水产品中硝基呋喃和硝基咪唑类药物残留量[J].理化检验-化学分册,2015,51(8):1168-1173.WAN J C,WANG W J,ZHAN C R,et al. HPLC-MS/MS simultaneous determination of residual amounts of nitrofuran and nitroimidazolesin aquatic products. PTCA(PART B:CHEM,ANAL.),2015,51(8):1168-1173(in Chinese).
[17] YU W H,CHIN T S,LAI H T. Detection of nitrofurans and their metabolites in pond water and sediments by liquid chromatography(LC)-photodiode array detection and LC-ion spray tandem mass spectrometry[J]. International Biodeterioration&Biodegradation,2013,85:517-526.
[18]王强,王旭峰,赵东豪,等.超高效液相色谱法测定水体和沉积物中4种硝基呋喃类抗生素[J].食品科学,2016,37(16):249-253.WANG Q,WANG X F,ZHAO D H,et al. Determination of four nitrofurans in water and sediment using ultra performance liquidchromatography[J]. Food Science,2016,37(16):249-253(in Chinese).
[19]尹怡,张宏峰,刘书贵,等.微波辅助衍生-液相色谱-串联质谱法快速测定养殖水体中5种硝基呋喃类代谢物残留[J].质谱学报,2016,37(2):106-113.YIN Y,ZHANG H F,LIU S G,et al. Dtermination of nitrofuran metabolites in fishery water by microwave assisted derivatization and liquidchromatography-tandem mass spectrometry[J]. Journal of Chinese Mass Spectrometry Society,2016,37(2):106-113(in Chinese).
[20]吴剑平,张婧,李丹妮,等.液相色谱串联质谱法正负模式切换同时检测饲料中10种硝基呋喃类化合物[J].中国兽药杂志,2017,21(11):51-58.WU J P,ZHANG J,LI D N,et al. Determination of 10 nitrofurans in feed by high performance liquid chromatography-tandem massspectrometry with positive and negative switching[J]. Chinese Journal of Veterinary Drug,2017,21(11):51-58(in Chinese).
[21]农业部783号公告-1-2006水产品中硝基呋喃类代谢物残留量的测定液相色谱-串联质谱法[S].Announcement of the Ministry of Agriculture 783-1-2006 Determination of nitrofuran metabolic residues in aquatic products by LC-MS/MSmethod[S](in Chinese).
[22] NOELIA M V T,PATRICIA P B,ROBERTO R G,et al. Determination of nitrofuran metabolites in seafood by ultra high performanceliquid chromatography coupled to triple quadrupole tandem mass spectrometry[J]. Journal of Food Composition and Analysis,2013,30:86-93.
[2] TAO Y F,YU H,CHEN D M,et al. Determination of sodium nifurstyrenate and nitrovin residues in edible food by liquid chromatography-tandem mass spectrometry after ultrasound-assisted extraction[J]. Journal of Chromatography B,2010,878:3415-3420.
[3]邢丽红,李兆新,孙伟红,等.液相色谱-串联质谱法检测水产品中硝基呋喃类药物的残留量[J].食品安全质量检测学报,2017,8(4):1233-1239.XING L H,LI Z X,SUN W H,et al. Simultaneous determination of four kinds of nitrofurans residues in aquatic products by liquidchromatography-tandem mass spectrometry[J]. Journal of Food Safety and Quality,2017,8(4):1233-1239(in Chinese).
[4] KAUFMANN A,BUTCHER P,MADEN K,et al. Determination of nitrofuran and chloramphenicol residues by high resolution massspectrometry versus tandem quadrupole mass spectrometry[J]. Analytica Chimica Acta,2015,862:41-52.
[5] VERDON E,COUEDOR P,SANDERS P. Multi-residue monitoring for the simultaneous determination of five nitrofurans(furazolidone,furaltadone,nitrofurazone,nitrofurantoine,nifursol)in poultry muscle tissue through the detection of their five major metabolites(AOZ,AMOZ,SEM,AHD,DNSAH)by liquid chromatography coupled to electrospray tandem mass spectrometry in house validation in line withcommission decision 657/2002/EC[J]. Analytica Chimica Acta,2007,586:336-347.
[6] KAUFAMANN A,BUTCHER P,MADEN K. LC-MS/MS method for determing nifursol and other nitrofuran residues in meat[J]. MittLebensm Hyg,2004,95:135-146.
[7]唐红梅,曾芳,李成洪.食品中硝基呋喃类药物及其代谢物残留检测的研究进展[J].食品安全质量检测学报,2016,7(10):3952-3954.TANG H M,ZENG F,LI C H. Progress on the detection of nitrofurans drugs residues and their metabolites in food[J]. Journal of FoodSafety and Quality,2016,7(10):3952-3954(in Chinese).
[8] FERNANDO R,MUNASINGHE D M S,GUNASENA A R C,et al. Determination of nitrofuran metabolites in shrimp muscle by liquidchromatography-photo diode array detection[J]. Food Control,2017,72:300-305.
[9] DU N N,CHEN M M,SHENG L Q,et al. Determination of nitrofuran metabolites in shrimp by high performance liquid chromatographywith fluorescence detection and liquid chromatography-tandem mass spectrometry using a new derivatization reagent[J]. Journal ofChromatography A,2014,1327:90-96.
[10] ZHANG Y B,QIAO H O,CHEN C,et al. Determination of nitrofurans metabolites residues in aquatic products by ultra-performance liquidchromatography-tandem mass spectrometry[J]. Food Chemistry,2016,192:612-617.
[11] ZHAI H Y,ZHANG L C,PAN Y F,et al. Simultaneous determination of chloramphenicol,ciprofloxacin,nitrofuran antibiotics and theirmetabolites in fishery products by CE[J]. Chromatographia,2015,78:551-556.
[12] JIANG W X,LUO P J,WANG X. Development of an enzyme-linked immunosorbent assay for the detection of nitrofurantoin metabolite,1-amino-hydantoin,in animal tissues[J]. Food Control,2012,23:20-25.
[13] ZHANG Z W,WU Y P,LI X W,et al. Multi-class method for the determination of nitroimidazoles,nitrofurans,and chloramphenicol inchicken muscle and egg by dispersive-solid phase extraction and ultra-high performance liquid chromatography-tandem mass spectrometry[J]. Food Chemistry,2017,217:182-190.
[14] DONGKYUM K,BYUNGJOO K,SEOK-WON H. An optimized method for the accurate determination of nitrofurans in chicken meat usingisotope dilution-liquid chromatography/mass spectrometry[J]. Journal of Food Composition and Analysis,2015,40:24-31.
[15]孙稚菁,王婵,李崇,等.高效液相色谱-串联质谱法快速检测海参中硝基呋喃代谢物前处理方案的探讨[J].分析测试学报,2016,35(9):1127-1131.SUN Z J,WANG C,LI C,et al. Pretreatment programs discussion of detecting nitrofuran metabolites rapidly in holothurians by highperformance liquid chromatography tandem mass spectrometry[J]. Journal of Instrumental Analysis,2016,35(9):1127-1131(inChinese).
[16]万建春,王文君,占春瑞,等.高效液相色谱-串联质谱法同时测定水产品中硝基呋喃和硝基咪唑类药物残留量[J].理化检验-化学分册,2015,51(8):1168-1173.WAN J C,WANG W J,ZHAN C R,et al. HPLC-MS/MS simultaneous determination of residual amounts of nitrofuran and nitroimidazolesin aquatic products. PTCA(PART B:CHEM,ANAL.),2015,51(8):1168-1173(in Chinese).
[17] YU W H,CHIN T S,LAI H T. Detection of nitrofurans and their metabolites in pond water and sediments by liquid chromatography(LC)-photodiode array detection and LC-ion spray tandem mass spectrometry[J]. International Biodeterioration&Biodegradation,2013,85:517-526.
[18]王强,王旭峰,赵东豪,等.超高效液相色谱法测定水体和沉积物中4种硝基呋喃类抗生素[J].食品科学,2016,37(16):249-253.WANG Q,WANG X F,ZHAO D H,et al. Determination of four nitrofurans in water and sediment using ultra performance liquidchromatography[J]. Food Science,2016,37(16):249-253(in Chinese).
[19]尹怡,张宏峰,刘书贵,等.微波辅助衍生-液相色谱-串联质谱法快速测定养殖水体中5种硝基呋喃类代谢物残留[J].质谱学报,2016,37(2):106-113.YIN Y,ZHANG H F,LIU S G,et al. Dtermination of nitrofuran metabolites in fishery water by microwave assisted derivatization and liquidchromatography-tandem mass spectrometry[J]. Journal of Chinese Mass Spectrometry Society,2016,37(2):106-113(in Chinese).
[20]吴剑平,张婧,李丹妮,等.液相色谱串联质谱法正负模式切换同时检测饲料中10种硝基呋喃类化合物[J].中国兽药杂志,2017,21(11):51-58.WU J P,ZHANG J,LI D N,et al. Determination of 10 nitrofurans in feed by high performance liquid chromatography-tandem massspectrometry with positive and negative switching[J]. Chinese Journal of Veterinary Drug,2017,21(11):51-58(in Chinese).
[21]农业部783号公告-1-2006水产品中硝基呋喃类代谢物残留量的测定液相色谱-串联质谱法[S].Announcement of the Ministry of Agriculture 783-1-2006 Determination of nitrofuran metabolic residues in aquatic products by LC-MS/MSmethod[S](in Chinese).
[22] NOELIA M V T,PATRICIA P B,ROBERTO R G,et al. Determination of nitrofuran metabolites in seafood by ultra high performanceliquid chromatography coupled to triple quadrupole tandem mass spectrometry[J]. Journal of Food Composition and Analysis,2013,30:86-93.