QuEChERS-高效液相色谱-串联质谱测定牛奶中6种玉米赤霉烯酮类毒素
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摘要
建立QuEChERS-高效液相色谱-串联质谱同时检测牛奶中6种玉米赤霉烯酮类毒素残留量的检测方法。样品用1%乙酸-乙腈提取,乙腈饱和正己烷除脂,C18、N-丙基乙二胺和无水硫酸镁净化,在ZORBAX SB-C18色谱柱上分离,以5 mmol/L NH4Ac溶液和乙腈作为流动相进行梯度洗脱,电喷雾离子源负离子模式扫描,多反应监测测定,基质匹配外标法定量分析。牛奶中6种玉米赤霉烯酮类毒素在0.1～50μg/L质量浓度范围内呈线性关系,相关系数均高于0.995;检出限为0.05～0.1μg/kg,定量限为0.25～0.5μg/kg。牛奶中6种玉米赤霉烯酮类毒素的3个加标水平的平均回收率范围为91.8%～114.5%,相对标准偏差在3.2%～14.4%之间(n=6)。
A method for simultaneous determination of six zeranols residues in milk was developed by quick, easy cheap,effective, rugged and safe(QuEChERS) extraction coupled to high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). The sample was extracted with 1% acetate acetonitrile. The extract was degreased with acetonitrile-saturated n-hexane, cleaned up with a mixture of C_(18), primary secondary amine(PSA) and magnesium sulfate anhydrous. The analytes were separated on a reversed phase C_(18) column and gradiently eluted with 5 mmol/L ammonium acetate and acetonitrile. These analytes were analyzed under the multiple reaction monitoring(MRM) mode with negative electrospray ionization(ESI-) and quanti?ed by matrix-matched external standard method. Good linearity was obtained for the six zeranols in the concentration range of 0.1–50 μg/L with correlation coef?cients of more than 0.995. The limits of detection(LODs) and the limits of quanti?cation(LOQs) for the zeranols were 0.05–0.1 and 0.25–0.5 μg/kg, respectively.The recoveries at three spiked concentration levels were in the range of 91.8%–114.5%. The repeatability expressed as relative standard deviation(RSD) ranged from 3.2% to 14.4%(n = 6).
A method for simultaneous determination of six zeranols residues in milk was developed by quick, easy cheap,effective, rugged and safe(QuEChERS) extraction coupled to high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). The sample was extracted with 1% acetate acetonitrile. The extract was degreased with acetonitrile-saturated n-hexane, cleaned up with a mixture of C_(18), primary secondary amine(PSA) and magnesium sulfate anhydrous. The analytes were separated on a reversed phase C_(18) column and gradiently eluted with 5 mmol/L ammonium acetate and acetonitrile. These analytes were analyzed under the multiple reaction monitoring(MRM) mode with negative electrospray ionization(ESI-) and quanti?ed by matrix-matched external standard method. Good linearity was obtained for the six zeranols in the concentration range of 0.1–50 μg/L with correlation coef?cients of more than 0.995. The limits of detection(LODs) and the limits of quanti?cation(LOQs) for the zeranols were 0.05–0.1 and 0.25–0.5 μg/kg, respectively.The recoveries at three spiked concentration levels were in the range of 91.8%–114.5%. The repeatability expressed as relative standard deviation(RSD) ranged from 3.2% to 14.4%(n = 6).
引文
[1]张天姝,马雷,玄兵,等.饲料中玉米赤霉醇、玉米赤霉酮和己烯雌酚的测定:超高效液相色谱-串联质谱法[J].现代畜牧兽医,2018(2):18-23.DOI:1672-9692(2018)02-0018-06.
[2]KUIPER-GOODMAN T,SCOTT P M,WATANABE H.Risk assessment of the mycotoxin zearalenone[J].Regulatory Toxicology&Pharmacology Rtp,1987,7(3):253-306.DOI:10.1016/0273-2300(87)90037-7.
[3]PAJEWSKA M,?OJKO M,CENDROWSKI K,et al.The determination of zearalenone and its major metabolites in endometrial cancer tissues[J].Analytical&Bioanalytical Chemistry,2018,410(12):1-12.DOI:10.1007/s00216-017-0807-7.
[4]国家卫生和计划生育委员会,国家食品药品监督管理总局.食品中真菌毒素限量:GB 2761-2017[S].北京:中国标准出版社,2017.
[5]王清,王国民,郗存显,等.复合免疫亲和柱净化-液相色谱-串联质谱法同时测定动物源性食品中6种玉米赤霉醇类化合物和氯霉素残留量[J].色谱,2014,32(6):640-646.DOI:10.3969/j.issn.1000-8713.2014.06.014.
[6]吴钟玲.食品中玉米赤霉醇及其类似物快速测定的研究[D].广州:华南理工大学,2011:4-5.
[7]佚名.玉米赤霉醇[J].质量探索,2013(增刊1):46.
[8]BELHASSEN H,JIMENEZ-DIAZ I,GHALI R,et al.Validation of a UHPLC-MS/MS method for quantification of zearalenone,alpha-zearalenol,beta-zearalenol,alphazearalanol,beta-zearalanol and zearalanone in human urine[J].Journal of Chromatography B,Analytical Technologies in the Biomedical and Life Sciences,2014;962:68-74.DOI:10.1016/j.jchromb.2014.05.019.
[9]ZINEDINE A,SORIANO J M,MOLT?J C,et al.Review on the toxicity,occurrence,metabolism,detoxification,regulations and intake of zearalenone:an oestrogenic mycotoxin[J].Food&Chemical Toxicology,2007,45(1):1-18.DOI:10.1016/j.fct.2006.07.030.
[10]农业部.动物性食品中兽药最高残留限量:农业部235号公告[S].北京:农业部,2012:1-26.
[11]LHOTSKáI,GAJDO?OVáB,SOLICH P,et al.Molecularly imprinted vs.reversed-phase extraction for the determination of zearalenone:a method development and critical comparison of sample clean-up efficiency achieved in an on-line coupled SPEchromatography system[J].Analytical&Bioanalytical Chemistry,2018,410(14):1-9.DOI:10.1007/s00216-018-0920-2.
[12]GUPTA R C,MOSTROM M S,EVANS T J.Zearalenone[M].Veterinary Toxicology.Academic Press,2018:1055-1063.
[13]LEEM J,KIMH J.Development of an immunoaf finity chromatography and LC-MS/MS method for the determination of 6zearalenones in animal feed[J].PLoS ONE,2018,13(3):e0193584.DOI:10.1371/journal.pone.0193584.
[14]朱捷,马桂娟,马雪梅.HPLC-MS/MS法测定牛奶中两种玉米赤霉醇[J].食品研究与开发,2017,38(6):159-163.DOI:10.3969/j.issn.1005-6521.2017.06.036.
[15]VISCONTI A,PASCALE M.Determination of zearalenone in corn by means of immunoaffinity clean-up and high-performance liquid chromatography with fluorescence detection[J].Journal of Chromatography A,1998,815(1):133-140.DOI:10.1016/S0021-9673(98)00296-9.
[16]李丽萍,范赛,赵榕,等.固相萃取-超高效液相色谱-串联质谱法检测猪肉中玉米赤霉醇类的残留量[J].色谱,2013,31(7):703-708.DOI:10.3724/SP.J.1123.2012.12033.
[17]邵瑞婷,张丽华,史娜,等.免疫亲和净化-超高效液相色谱-串联质谱法测定食品中玉米赤霉烯酮类真菌毒素[J].食品科学,2017,38(16):274-279.DOI:10.7506/spkx1002-6630-201716044.
[18]YAN Z,WANG L,WANG J,et al.A QuEChERS-based liquid chromatography-tandem mass spectrometry method for the simultaneous determination of nine zearalenone-like mycotoxins in pigs[J].Toxins,2018,10(3):129.DOI:10.3390/toxins10030129.
[19]甄振鹏,高裕锋,张琳,等.QuEChERS-高效液相色谱串联质谱法测定牛奶中6种玉米赤霉醇类激素药物残留量[J].广东化工,2016,43(13):256-258.DOI:10.3969/j.issn.1007-1865.2016.13.125.
[20]国家卫生和计划生育委员会,国家食品药品监督管理总局食品中玉米赤霉烯酮的测定:GB 5009.209-2016[S].北京:中国标准出版社,2016:7-11.
[21]钱卓真,刘智禹,邓武剑,等.高效液相色谱-串联质谱法测定水产品中玉米赤霉醇类激素药物残留量[J].南方水产科学,2011,7(1):62-68.DOI:10.3969/j.issn.2095-0780.2011.01.010.
[22]彭涛,李晓娟,陈冬东,等.高效液相色谱-串联质谱法同时测定动物肝脏中玉米赤霉醇及其类似物残留量[J].分析化学,2010,38(4):469-474.DOI:10.3724/SP.J.1096.2010.00469.
[23]方晓明,陈家华,唐毅锋.高效液相色谱法测定鸡肝中玉米赤霉醇的残留量[J].色谱,2003,21(2):158-161.DOI:10.3321/j.issn:1000-8713.2003.02.015.
[24]王炼,王希希,张新申.同位素稀释-液相色谱-串联质谱法测定动物源性食品中6种玉米赤霉醇类化合物[J].中国食品卫生杂志,2015,27(5):530-534.DOI:10.13590/j.cjfh.2015.05.010.
[25]游丽娜,李贤良,郗存显,等.免疫亲和柱净化-高效液相色谱法同时检测鸡蛋中6种玉米赤霉醇类化合物残留量[J].色谱,2012,30(10):1021-1025.DOI:10.3724/SP.J.1123.2012.08016.
[26]BERTHILLER F,SCHUHMACHER R,BUTTINGER G,et al.Rapid simultaneous determination of major type A-and B-trichothecenes as well as zearalenone in maize by high performance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2005,1062(2):209-216.DOI:10.1016/j.chroma.2004.11.011.
[27]李贤良,游丽娜,郗存显,等.免疫亲和柱净化-液相色谱-串联质谱法同时测定猪肉中6种玉米赤霉醇类化合物残留量[J].分析化学,2013,41(8):1147-1152.DOI:10.3724/SP.J.1096.2013.30141.
[28]王昕.IAC-LC法和IAC-LC-MS/MS法同时检测食品中黄曲霉毒素及玉米赤霉醇类残留量[D].杨凌:西北农林科技大学,2014:28-29.
[29]农业部.动物源性食品中玉米赤霉醇类药物残留检测液相色谱-串联质谱法:农业部1025号公告-19-2008[S].2008.
[30]谢继安,刘柏林.超高压液相色谱-串联质谱法检测猪肉中玉米赤霉醇类物质[J].食品安全质量检测学报,2013,4(1):150-158.
[2]KUIPER-GOODMAN T,SCOTT P M,WATANABE H.Risk assessment of the mycotoxin zearalenone[J].Regulatory Toxicology&Pharmacology Rtp,1987,7(3):253-306.DOI:10.1016/0273-2300(87)90037-7.
[3]PAJEWSKA M,?OJKO M,CENDROWSKI K,et al.The determination of zearalenone and its major metabolites in endometrial cancer tissues[J].Analytical&Bioanalytical Chemistry,2018,410(12):1-12.DOI:10.1007/s00216-017-0807-7.
[4]国家卫生和计划生育委员会,国家食品药品监督管理总局.食品中真菌毒素限量:GB 2761-2017[S].北京:中国标准出版社,2017.
[5]王清,王国民,郗存显,等.复合免疫亲和柱净化-液相色谱-串联质谱法同时测定动物源性食品中6种玉米赤霉醇类化合物和氯霉素残留量[J].色谱,2014,32(6):640-646.DOI:10.3969/j.issn.1000-8713.2014.06.014.
[6]吴钟玲.食品中玉米赤霉醇及其类似物快速测定的研究[D].广州:华南理工大学,2011:4-5.
[7]佚名.玉米赤霉醇[J].质量探索,2013(增刊1):46.
[8]BELHASSEN H,JIMENEZ-DIAZ I,GHALI R,et al.Validation of a UHPLC-MS/MS method for quantification of zearalenone,alpha-zearalenol,beta-zearalenol,alphazearalanol,beta-zearalanol and zearalanone in human urine[J].Journal of Chromatography B,Analytical Technologies in the Biomedical and Life Sciences,2014;962:68-74.DOI:10.1016/j.jchromb.2014.05.019.
[9]ZINEDINE A,SORIANO J M,MOLT?J C,et al.Review on the toxicity,occurrence,metabolism,detoxification,regulations and intake of zearalenone:an oestrogenic mycotoxin[J].Food&Chemical Toxicology,2007,45(1):1-18.DOI:10.1016/j.fct.2006.07.030.
[10]农业部.动物性食品中兽药最高残留限量:农业部235号公告[S].北京:农业部,2012:1-26.
[11]LHOTSKáI,GAJDO?OVáB,SOLICH P,et al.Molecularly imprinted vs.reversed-phase extraction for the determination of zearalenone:a method development and critical comparison of sample clean-up efficiency achieved in an on-line coupled SPEchromatography system[J].Analytical&Bioanalytical Chemistry,2018,410(14):1-9.DOI:10.1007/s00216-018-0920-2.
[12]GUPTA R C,MOSTROM M S,EVANS T J.Zearalenone[M].Veterinary Toxicology.Academic Press,2018:1055-1063.
[13]LEEM J,KIMH J.Development of an immunoaf finity chromatography and LC-MS/MS method for the determination of 6zearalenones in animal feed[J].PLoS ONE,2018,13(3):e0193584.DOI:10.1371/journal.pone.0193584.
[14]朱捷,马桂娟,马雪梅.HPLC-MS/MS法测定牛奶中两种玉米赤霉醇[J].食品研究与开发,2017,38(6):159-163.DOI:10.3969/j.issn.1005-6521.2017.06.036.
[15]VISCONTI A,PASCALE M.Determination of zearalenone in corn by means of immunoaffinity clean-up and high-performance liquid chromatography with fluorescence detection[J].Journal of Chromatography A,1998,815(1):133-140.DOI:10.1016/S0021-9673(98)00296-9.
[16]李丽萍,范赛,赵榕,等.固相萃取-超高效液相色谱-串联质谱法检测猪肉中玉米赤霉醇类的残留量[J].色谱,2013,31(7):703-708.DOI:10.3724/SP.J.1123.2012.12033.
[17]邵瑞婷,张丽华,史娜,等.免疫亲和净化-超高效液相色谱-串联质谱法测定食品中玉米赤霉烯酮类真菌毒素[J].食品科学,2017,38(16):274-279.DOI:10.7506/spkx1002-6630-201716044.
[18]YAN Z,WANG L,WANG J,et al.A QuEChERS-based liquid chromatography-tandem mass spectrometry method for the simultaneous determination of nine zearalenone-like mycotoxins in pigs[J].Toxins,2018,10(3):129.DOI:10.3390/toxins10030129.
[19]甄振鹏,高裕锋,张琳,等.QuEChERS-高效液相色谱串联质谱法测定牛奶中6种玉米赤霉醇类激素药物残留量[J].广东化工,2016,43(13):256-258.DOI:10.3969/j.issn.1007-1865.2016.13.125.
[20]国家卫生和计划生育委员会,国家食品药品监督管理总局食品中玉米赤霉烯酮的测定:GB 5009.209-2016[S].北京:中国标准出版社,2016:7-11.
[21]钱卓真,刘智禹,邓武剑,等.高效液相色谱-串联质谱法测定水产品中玉米赤霉醇类激素药物残留量[J].南方水产科学,2011,7(1):62-68.DOI:10.3969/j.issn.2095-0780.2011.01.010.
[22]彭涛,李晓娟,陈冬东,等.高效液相色谱-串联质谱法同时测定动物肝脏中玉米赤霉醇及其类似物残留量[J].分析化学,2010,38(4):469-474.DOI:10.3724/SP.J.1096.2010.00469.
[23]方晓明,陈家华,唐毅锋.高效液相色谱法测定鸡肝中玉米赤霉醇的残留量[J].色谱,2003,21(2):158-161.DOI:10.3321/j.issn:1000-8713.2003.02.015.
[24]王炼,王希希,张新申.同位素稀释-液相色谱-串联质谱法测定动物源性食品中6种玉米赤霉醇类化合物[J].中国食品卫生杂志,2015,27(5):530-534.DOI:10.13590/j.cjfh.2015.05.010.
[25]游丽娜,李贤良,郗存显,等.免疫亲和柱净化-高效液相色谱法同时检测鸡蛋中6种玉米赤霉醇类化合物残留量[J].色谱,2012,30(10):1021-1025.DOI:10.3724/SP.J.1123.2012.08016.
[26]BERTHILLER F,SCHUHMACHER R,BUTTINGER G,et al.Rapid simultaneous determination of major type A-and B-trichothecenes as well as zearalenone in maize by high performance liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2005,1062(2):209-216.DOI:10.1016/j.chroma.2004.11.011.
[27]李贤良,游丽娜,郗存显,等.免疫亲和柱净化-液相色谱-串联质谱法同时测定猪肉中6种玉米赤霉醇类化合物残留量[J].分析化学,2013,41(8):1147-1152.DOI:10.3724/SP.J.1096.2013.30141.
[28]王昕.IAC-LC法和IAC-LC-MS/MS法同时检测食品中黄曲霉毒素及玉米赤霉醇类残留量[D].杨凌:西北农林科技大学,2014:28-29.
[29]农业部.动物源性食品中玉米赤霉醇类药物残留检测液相色谱-串联质谱法:农业部1025号公告-19-2008[S].2008.
[30]谢继安,刘柏林.超高压液相色谱-串联质谱法检测猪肉中玉米赤霉醇类物质[J].食品安全质量检测学报,2013,4(1):150-158.