液相色谱-串联质谱法测定鸡蛋中氟虫腈及其代谢物的残留量
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摘要
建立一种采用液相色谱-三重四级杆质谱测定鸡蛋中氟虫腈及其代谢物(氟甲腈、氟虫腈砜和氟虫腈硫醚)的快速筛查方法。鸡蛋样品经乙腈和正己烷提取,C18固相萃取柱净化,Shim-pack XR-ODSⅢ色谱柱(2.0 mm i.D×150 mm,2.1μm)分离,以乙腈-5 mmol/L乙酸铵(0.02%甲酸)水溶液为流动相,梯度洗脱,外标法定量。采用液相色谱-三重四级杆串联质谱动态多反应监测模式,以负离子采集进行定性筛查和定量分析。结果表明,氟虫腈及其代谢物在0～50μg/L质量浓度范围内线性关系良好,相关系数R2均大于0.999。以10倍信噪比确定各药物的定量限(LOQ),氟虫腈及其代谢物的定量限为0.3～0.6μg/kg。在0.5、5.0和10.0μg/kg添加水平下,氟虫腈及其代谢物的平均回收率为92.3%～105.1%,相对标准偏差(RSD)为1.2%～8.6%。该方法简便、快速、灵敏,适用于鸡蛋中氟虫腈及其代谢物的快速筛查和定量检测。
A rapid screening method was developed for the determination of the residues of fipronil and its metabolites(fipronil-desulfinyl,fipronil sulfone and fiproil sulfide) in eggs by liquid chromatography triple quadrupole mass spectrometry(LC-MS/MS).Egg samples were extracted with acetonitrile and n-hexane.The obtained extract was purified by a C18 solid phase extraction(SPE) column and further separated using a Shim-pack XR-ODS columnⅢ(2.0 mm I.D.× 150 mmL.,2.1 μm particle size) with a mobile phase in gradient elution(acetonitrile-5 mmol/L ammonium acetate(0.02% formic acid).Qualitative screening and quantitative analysis were performed by LC-MS/MS with dynamic multiple reaction monitoring(DMRM) mode and in negative ion mode,and quantification was carried out by the external standard method.The results showed that good linearity was detected for fipronil and its metabolites in the range of 0～50 μg/L,with the correlation coefficient R2 greater than 0.999.The limit of quantification(LOQ) of fipronil and its metabolites was 0.3～0.6 μg/kg at the signal-to-noise ratio of 10.The average recovery of fipronil and its metabolites at the spiking levels of 0.5,5.0 and 10.0 μg/kg ranged from 92.3% to 105.1%,with the relative standard deviation(RSD,n=6) in the range of 1.2%～8.6% The method was proven to be simple,rapid andsensitive,and suitable for rapid screening and quantitative detection of fipronil and metabolite residues in eggs.
A rapid screening method was developed for the determination of the residues of fipronil and its metabolites(fipronil-desulfinyl,fipronil sulfone and fiproil sulfide) in eggs by liquid chromatography triple quadrupole mass spectrometry(LC-MS/MS).Egg samples were extracted with acetonitrile and n-hexane.The obtained extract was purified by a C18 solid phase extraction(SPE) column and further separated using a Shim-pack XR-ODS columnⅢ(2.0 mm I.D.× 150 mmL.,2.1 μm particle size) with a mobile phase in gradient elution(acetonitrile-5 mmol/L ammonium acetate(0.02% formic acid).Qualitative screening and quantitative analysis were performed by LC-MS/MS with dynamic multiple reaction monitoring(DMRM) mode and in negative ion mode,and quantification was carried out by the external standard method.The results showed that good linearity was detected for fipronil and its metabolites in the range of 0～50 μg/L,with the correlation coefficient R2 greater than 0.999.The limit of quantification(LOQ) of fipronil and its metabolites was 0.3～0.6 μg/kg at the signal-to-noise ratio of 10.The average recovery of fipronil and its metabolites at the spiking levels of 0.5,5.0 and 10.0 μg/kg ranged from 92.3% to 105.1%,with the relative standard deviation(RSD,n=6) in the range of 1.2%～8.6% The method was proven to be simple,rapid andsensitive,and suitable for rapid screening and quantitative detection of fipronil and metabolite residues in eggs.
引文
[1]操海群,施艳红,花日茂.辣椒,白菜中氟虫腈残留分析方法的研究[J].食品科学,2005,26(9):377-380CAO Hai-qun,SHI Yan-hong,HUA Ri-mao,et al.Study on analytical method on fipronil residue in pepper and Chinese Cabbage[J].Food Science,2005,26(9):377-380
[2]吕冰,尹帅星,陈达炜,等.Qu Ech ERS-四级杆/静电场轨道阱高分辨质谱测定动物性食品中氟虫腈及其代谢物残留[J].分析测试学报,2017,36(12):1424-1430LYU Bing,YIN Shuai-xing,CHEN Da-wei,et al.Determination of Fipronil and its metabolites in animal derived foods by Qu ECh ERS method combined with ultra performance liquid chromatography-high-resolution benchtop Q exactive mass spectrometry[J].Journal of Instrumental Analysis,2017,36(12):1424-1430
[3]白宝清,李美萍,张生万.Qu ECh ERS-DLLME-高效液相色谱法测定蔬菜中溴虫腈和氟虫腈残留[J].食品科学,2014,35(24):254-258BAI Bao-qing,LI Mei-ping,ZHANG Sheng-wan,et al.Determination of chlorfenapyr and fipronil residues in vegetables by Qu ECh ERS combined with dispersive liquid-liquid microextraction and high performance liquid chromatography[J].Food Science,2014,35(24):254-258
[4]周昱,徐敦明,陈达捷,等.固相微萃取-气相色谱法和气相色谱-质谱法测定茶叶中氟虫腈及其代谢物残留[J].色谱,2011,29(7):656-661ZHOU Yu,XU Dun-ming,CHEN Da-jie,et al.Determination of fipronil and its metabolites in tea by solid-phase microextraction coupled with gas chromatography and gas chromatography-mass spectrometry[J].Chinese Journal of Chromatography,2011,29(7):656-661
[5]Kurz M H S,Martel S,Goncalves F,et al.Development of a fast method for the determination of the insecticide fipronil and its metabolites in environmental waters by SPE and GC-ECD[J].Journal of the Brazilian Chemical Society,2013,24(4):631-638
[6]CAO Yong song,CHEN Jiu xin,WANG Yue long,et al.HPLC/UV analysis of chlorfenapyr residues in cabbage and soil to study the dynamics of different formulations[J].Science of the Total Environment,2005,350(1/3):38-46
[7]Tomasini D,Sampaio M R F,Cardoso L V,et al.Comparison of dispersive liquid-liquid microextraction and the modified Qu ECh ERS method for the determination of fipronil in honey by high performance liquid chromatography with diode-array detection[J].Analytical Methods,2011,3(8):1893-1900
[8]Hadjmohammadi M R,Nikou S M,Kamel K.Determination of fipronil residue in soil and water in the rice fields in north of Iran by RP-HPLC method[J].Acta Chimica Slovenica,2006,53(4):517-520
[9]Cid Y P,Ferreira T P,Medeiros D M V C,et al.Determination of fipronil in bovine plasma by solid phase extraction and liquid chromatography with ultraviolet detection[J].Quimica Nova,2010,35(10):2063-2066
[10]Ramasubramanian T,Paramasivam M,Jayanthi R,et al.Asimple and sensitive single-step method for gas chromatography-mass spectrometric determination of fipronil and its metabolites in sugarcane juice,jaggery and sugar[J].Food Chemistry,2014,150:408-413
[11]Paramasivam M.Chandrasekaran S.Determination of fipronil and its major metabolites in vegetables,fruit and soil using Qu ECh ERS and gas chromatography-mass spectrometry[J].International Journal of Environmental Analytical Chemistry.2013,93(11):1203-1211
[12]Araujo L,Troconis M,Cubillan D,et al.Single drop microextraction and gas chromatography-mass spectrometry for the determination of diflufenican.mepanipyrim,fipronil,and pretilachlor in water samples[J].Environmental Monitoring and Assessment.2013,185(12):10225-10233
[13]Sartarelli N C,De Macedo A N,De Sousa J P,et al.Determination of chlorfenvinphos,fipronil,and cypermethrin residues in meat and bovine fat using Qu ECh ERS method and gas chromatography-mass spectrometry[J].Journal of Liquid Chromatography and Related Technologies,2012,35(13):1895-1908
[14]薄海波.气相色谱-串联质谱正离子模式测定食品中氟虫腈和溴虫腈残留[J].中国食品卫生杂志,2013,25(5):431-434BO Hai-bo.Determination of fipronil and chlorfenapyr residues in foodstuffs by gas chromatography-tandem mass spectrometry in positive ion mode[J].Chinese Journal of Food Hygiene,2013,25(5):431-434
[15]Débora T,Maicon R F.S,Sergiane S C,et al.Simultaneous determination of pesticides and 5-liydroxymetliylfurfural in honey by the modified Qu ECh ERS method and liquid chromatography coupled to tandem mass spectrometry[J].Talanta,2012,99:380-386
[16]丁明,钟冬莲,汤富彬,等.固相萃取-高效液相色谱-串联质谱联用测定竹笋中残留的7种杀虫剂农药[J].色谱,2013,31(2):117-121DING Ming,ZHONG Dong-lian,TANG Fu-bin,et al.Determination of seven pesticide residues in bamboo shoots by high performance liquid chromatography-tandem mass spectrometry coupled with solid-phase extraction[J].Chinese Journal of Chromatography,2013,31(2):117-121
[17]Sandip H,Dasharath P O,Sagar C U,et al.Residue analysis of fipronil and difenoconazole in okra by liquid chromatography tandem mass spectrometry and their food safety evaluation[J].Food Chemistry,2015,176:145-151
[18]Maria G,Maria J A,Gonzalo F,et al.Validation of an off line solid phase extraction liquid chromatography-tandem mass spectrometry method for the determination of systemic insecticide residues in honey and pollen samples collected in apiaries from NW Spain[J].Analytica Chimica Acta,2010,672(1-2):107-113
[19]堵燕饪,罗漪涟,王洁琼,等.在线净化-液相色谱串联质谱技术测定蔬菜中氟虫腈及其代谢物残留[J].环境化学,2017,36(4):928-930DU Yan-yu,LUO Yi-lian,WANG Jie-qiong,et al.Determination of fipronil and its metabolites in vegetables by on-line cleanup liquid chromatography-tandem mass spectrometry[J].Environmental Chemistry,2017,36(4):928-930
[20]胡贝贞,蔡海江,宋伟华.茶叶中氟虫睛等8种农药残留的液相色谱-串联质谱法测定及不确定度评定[J].色谱,2012,30(9):889-895HU Bei-zhen,CAI Hai-jiang,SONG Wei-hua.Determination of eight pesticide residues in tea by liquid chromatography-tandem mass spectrometry and its uncertainty evaluation[J].Chinese Journal of Chromatography,2012,30(9):889-895
[21]吴成,任海雷,赵志强,等.液相色谱-串联质谱法检测芹菜中氟虫睛及其代谢物残留[J].福建分析测试,2016,25(5):37-40WU Cheng,REN Hai-lei,ZHAO Zhi-qiang,et al.Determination of the fipronil and its metabolites residues in celery by liquid chromatography/tandem mass spectrometry[J].Fujian Analysis&Testing,2016,25(5):37-40
[22]向平,沈敏,卓先义.液相色谱-质谱联用技术在药物和毒物分析中的应用[M].上海:上海科学技术出版社,2009:21-26XIANG Ping,SHEN Min,ZHUO Xian-yi.Application of liquid chromatography-mass spectrometry in the analysis of drugs and toxicants[M].Shanghai:Shanghai Scientific&Technical Publishers,2009:21-26
[23]贺敏,宋丹,董丰收,等.UPLC-MS/MS快速检测韭菜和土壤中氟虫腈及其代谢物残留[J].环境化学,2016,35(5):925-932HE Min,SONG Dan,DONG Feng-shou,et al.Determination of fipronil and its metabolites in leek and soil by ultrahigh performance liquid chromatography-mass spectrometry[J].Environmental Chemistry,2016,35(5):925-932
[24]Layne J,Farcas T,Rustamov I,et al.Volume-load capacity in fast-gradient liquid chromatography:effect of sample solvent composition and injection volume on chromatographic performance[J].Journal of Chromatography A,2001,913(1):233-242
[25]Patel D.Matrix effect in a view of LC-MS/MS:An overview[J].International Journal of Pharma and Bio Sciences,2011,2:559-564
[26]王建华,葛敏敏,张慧丽,等.Qu ECh ERS法-液相色谱-串联质谱法测定蔬菜中7种农药及其代谢物的残留量[J].食品科学,2017,38(20):235-241WANG Jian-hua,GE Min-min,ZHANG Hui-li,et al.Determination of 7 pesticide and metabolite residues in vegetables using Qu ECh ERS and liquid chromatography-tandem mass spectrometry[J].Food Science,2017,38(20):235-241
[27]Matusewski B K,Constanzer M L,Chavez-eng C M.Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS[J].Analytical Chemistry,2003,75(13):3019-3030
[28]GB/T 27404-2008,实验室质量控制规范食品理化检验[S].GB/T 27404-2008,Criterion on Quality Control of Laboratories-chemical Testing of Food[S]
[2]吕冰,尹帅星,陈达炜,等.Qu Ech ERS-四级杆/静电场轨道阱高分辨质谱测定动物性食品中氟虫腈及其代谢物残留[J].分析测试学报,2017,36(12):1424-1430LYU Bing,YIN Shuai-xing,CHEN Da-wei,et al.Determination of Fipronil and its metabolites in animal derived foods by Qu ECh ERS method combined with ultra performance liquid chromatography-high-resolution benchtop Q exactive mass spectrometry[J].Journal of Instrumental Analysis,2017,36(12):1424-1430
[3]白宝清,李美萍,张生万.Qu ECh ERS-DLLME-高效液相色谱法测定蔬菜中溴虫腈和氟虫腈残留[J].食品科学,2014,35(24):254-258BAI Bao-qing,LI Mei-ping,ZHANG Sheng-wan,et al.Determination of chlorfenapyr and fipronil residues in vegetables by Qu ECh ERS combined with dispersive liquid-liquid microextraction and high performance liquid chromatography[J].Food Science,2014,35(24):254-258
[4]周昱,徐敦明,陈达捷,等.固相微萃取-气相色谱法和气相色谱-质谱法测定茶叶中氟虫腈及其代谢物残留[J].色谱,2011,29(7):656-661ZHOU Yu,XU Dun-ming,CHEN Da-jie,et al.Determination of fipronil and its metabolites in tea by solid-phase microextraction coupled with gas chromatography and gas chromatography-mass spectrometry[J].Chinese Journal of Chromatography,2011,29(7):656-661
[5]Kurz M H S,Martel S,Goncalves F,et al.Development of a fast method for the determination of the insecticide fipronil and its metabolites in environmental waters by SPE and GC-ECD[J].Journal of the Brazilian Chemical Society,2013,24(4):631-638
[6]CAO Yong song,CHEN Jiu xin,WANG Yue long,et al.HPLC/UV analysis of chlorfenapyr residues in cabbage and soil to study the dynamics of different formulations[J].Science of the Total Environment,2005,350(1/3):38-46
[7]Tomasini D,Sampaio M R F,Cardoso L V,et al.Comparison of dispersive liquid-liquid microextraction and the modified Qu ECh ERS method for the determination of fipronil in honey by high performance liquid chromatography with diode-array detection[J].Analytical Methods,2011,3(8):1893-1900
[8]Hadjmohammadi M R,Nikou S M,Kamel K.Determination of fipronil residue in soil and water in the rice fields in north of Iran by RP-HPLC method[J].Acta Chimica Slovenica,2006,53(4):517-520
[9]Cid Y P,Ferreira T P,Medeiros D M V C,et al.Determination of fipronil in bovine plasma by solid phase extraction and liquid chromatography with ultraviolet detection[J].Quimica Nova,2010,35(10):2063-2066
[10]Ramasubramanian T,Paramasivam M,Jayanthi R,et al.Asimple and sensitive single-step method for gas chromatography-mass spectrometric determination of fipronil and its metabolites in sugarcane juice,jaggery and sugar[J].Food Chemistry,2014,150:408-413
[11]Paramasivam M.Chandrasekaran S.Determination of fipronil and its major metabolites in vegetables,fruit and soil using Qu ECh ERS and gas chromatography-mass spectrometry[J].International Journal of Environmental Analytical Chemistry.2013,93(11):1203-1211
[12]Araujo L,Troconis M,Cubillan D,et al.Single drop microextraction and gas chromatography-mass spectrometry for the determination of diflufenican.mepanipyrim,fipronil,and pretilachlor in water samples[J].Environmental Monitoring and Assessment.2013,185(12):10225-10233
[13]Sartarelli N C,De Macedo A N,De Sousa J P,et al.Determination of chlorfenvinphos,fipronil,and cypermethrin residues in meat and bovine fat using Qu ECh ERS method and gas chromatography-mass spectrometry[J].Journal of Liquid Chromatography and Related Technologies,2012,35(13):1895-1908
[14]薄海波.气相色谱-串联质谱正离子模式测定食品中氟虫腈和溴虫腈残留[J].中国食品卫生杂志,2013,25(5):431-434BO Hai-bo.Determination of fipronil and chlorfenapyr residues in foodstuffs by gas chromatography-tandem mass spectrometry in positive ion mode[J].Chinese Journal of Food Hygiene,2013,25(5):431-434
[15]Débora T,Maicon R F.S,Sergiane S C,et al.Simultaneous determination of pesticides and 5-liydroxymetliylfurfural in honey by the modified Qu ECh ERS method and liquid chromatography coupled to tandem mass spectrometry[J].Talanta,2012,99:380-386
[16]丁明,钟冬莲,汤富彬,等.固相萃取-高效液相色谱-串联质谱联用测定竹笋中残留的7种杀虫剂农药[J].色谱,2013,31(2):117-121DING Ming,ZHONG Dong-lian,TANG Fu-bin,et al.Determination of seven pesticide residues in bamboo shoots by high performance liquid chromatography-tandem mass spectrometry coupled with solid-phase extraction[J].Chinese Journal of Chromatography,2013,31(2):117-121
[17]Sandip H,Dasharath P O,Sagar C U,et al.Residue analysis of fipronil and difenoconazole in okra by liquid chromatography tandem mass spectrometry and their food safety evaluation[J].Food Chemistry,2015,176:145-151
[18]Maria G,Maria J A,Gonzalo F,et al.Validation of an off line solid phase extraction liquid chromatography-tandem mass spectrometry method for the determination of systemic insecticide residues in honey and pollen samples collected in apiaries from NW Spain[J].Analytica Chimica Acta,2010,672(1-2):107-113
[19]堵燕饪,罗漪涟,王洁琼,等.在线净化-液相色谱串联质谱技术测定蔬菜中氟虫腈及其代谢物残留[J].环境化学,2017,36(4):928-930DU Yan-yu,LUO Yi-lian,WANG Jie-qiong,et al.Determination of fipronil and its metabolites in vegetables by on-line cleanup liquid chromatography-tandem mass spectrometry[J].Environmental Chemistry,2017,36(4):928-930
[20]胡贝贞,蔡海江,宋伟华.茶叶中氟虫睛等8种农药残留的液相色谱-串联质谱法测定及不确定度评定[J].色谱,2012,30(9):889-895HU Bei-zhen,CAI Hai-jiang,SONG Wei-hua.Determination of eight pesticide residues in tea by liquid chromatography-tandem mass spectrometry and its uncertainty evaluation[J].Chinese Journal of Chromatography,2012,30(9):889-895
[21]吴成,任海雷,赵志强,等.液相色谱-串联质谱法检测芹菜中氟虫睛及其代谢物残留[J].福建分析测试,2016,25(5):37-40WU Cheng,REN Hai-lei,ZHAO Zhi-qiang,et al.Determination of the fipronil and its metabolites residues in celery by liquid chromatography/tandem mass spectrometry[J].Fujian Analysis&Testing,2016,25(5):37-40
[22]向平,沈敏,卓先义.液相色谱-质谱联用技术在药物和毒物分析中的应用[M].上海:上海科学技术出版社,2009:21-26XIANG Ping,SHEN Min,ZHUO Xian-yi.Application of liquid chromatography-mass spectrometry in the analysis of drugs and toxicants[M].Shanghai:Shanghai Scientific&Technical Publishers,2009:21-26
[23]贺敏,宋丹,董丰收,等.UPLC-MS/MS快速检测韭菜和土壤中氟虫腈及其代谢物残留[J].环境化学,2016,35(5):925-932HE Min,SONG Dan,DONG Feng-shou,et al.Determination of fipronil and its metabolites in leek and soil by ultrahigh performance liquid chromatography-mass spectrometry[J].Environmental Chemistry,2016,35(5):925-932
[24]Layne J,Farcas T,Rustamov I,et al.Volume-load capacity in fast-gradient liquid chromatography:effect of sample solvent composition and injection volume on chromatographic performance[J].Journal of Chromatography A,2001,913(1):233-242
[25]Patel D.Matrix effect in a view of LC-MS/MS:An overview[J].International Journal of Pharma and Bio Sciences,2011,2:559-564
[26]王建华,葛敏敏,张慧丽,等.Qu ECh ERS法-液相色谱-串联质谱法测定蔬菜中7种农药及其代谢物的残留量[J].食品科学,2017,38(20):235-241WANG Jian-hua,GE Min-min,ZHANG Hui-li,et al.Determination of 7 pesticide and metabolite residues in vegetables using Qu ECh ERS and liquid chromatography-tandem mass spectrometry[J].Food Science,2017,38(20):235-241
[27]Matusewski B K,Constanzer M L,Chavez-eng C M.Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS[J].Analytical Chemistry,2003,75(13):3019-3030
[28]GB/T 27404-2008,实验室质量控制规范食品理化检验[S].GB/T 27404-2008,Criterion on Quality Control of Laboratories-chemical Testing of Food[S]