不同净化柱处理对小麦粉中4种镰刀菌毒素的净化效果
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摘要
比较MycoSep~?226多功能净化柱、DZT MS-PREP免疫亲和柱和实验室制备聚苯乙烯-二乙烯苯树脂免疫亲和柱在检测小麦粉中4种镰刀菌毒素(玉米赤霉烯酮、脱氧雪腐镰刀菌烯醇、T-2毒素、HT-2毒素)过程中的前处理效果。选择MycoSep~?226多功能净化柱、DZT MS-PREP免疫亲和柱和实验室制备聚苯乙烯-二乙烯苯树脂免疫亲和柱,通过不同的提取和前处理方法,以及建立高效液相色谱-串联质谱检测法,对小麦粉中的4种镰刀菌毒素进行测定。DZT MS-PREP免疫亲和柱处理小麦粉样品所得的回收率为88.01%~107.31%,相对标准偏差为1.09%~14.42%;聚苯乙烯-二乙烯苯树脂免疫亲和柱的回收率为70.05%~112.80%,相对标准偏差为3.42%~12.25%;MycoSep~?226多功能净化柱的回收率为69.42%~111.12%,相对标准偏差为1.46%~13.24%。免疫亲和柱具有重复利用,有机溶剂耗用量少、特异性强等显著优势,且聚苯乙烯-二乙烯苯树脂免疫亲和柱为新载体免疫亲和柱的制备提供了依据。
A method for the simultaneous determination of four Fusarium toxins in wheat flour including zenralenone, deoxynivalenol, T-2 and HT-2 toxins was established using high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Samples were cleaned up with a MycoSep~?226 multifunctional column, a DZT MS-PREP immunoaffinity column, which were both commercial products, and a polystyrene-divinylbenzene(PS-DVB) immunoaffinity column which was prepared in our laboratory, respectively. The recoveries of Fusarium toxins purified with MycoSep~?226 ranged from 69.42% to 111.12% with relative standard deviations(RSDs) of 1.46%–13.24%, those obtained with PS-DVB ranged from 70.05% to 112.80% with RSDs of 3.42%–12.25%, and those obtained with DZT MS-PREP ranged from 88.01% to 107.31% with RSDs of 1.09%–14.42%. Immunoaffinity columns had the advantages of repeated use, less organic solvent consumption and high specificity. Moreover, PS-DVB could be a promising new immunoaffinity carrier.
A method for the simultaneous determination of four Fusarium toxins in wheat flour including zenralenone, deoxynivalenol, T-2 and HT-2 toxins was established using high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS). Samples were cleaned up with a MycoSep~?226 multifunctional column, a DZT MS-PREP immunoaffinity column, which were both commercial products, and a polystyrene-divinylbenzene(PS-DVB) immunoaffinity column which was prepared in our laboratory, respectively. The recoveries of Fusarium toxins purified with MycoSep~?226 ranged from 69.42% to 111.12% with relative standard deviations(RSDs) of 1.46%–13.24%, those obtained with PS-DVB ranged from 70.05% to 112.80% with RSDs of 3.42%–12.25%, and those obtained with DZT MS-PREP ranged from 88.01% to 107.31% with RSDs of 1.09%–14.42%. Immunoaffinity columns had the advantages of repeated use, less organic solvent consumption and high specificity. Moreover, PS-DVB could be a promising new immunoaffinity carrier.
引文
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[12]孙娟,李为喜,张妍,等.用超高效液相色谱串联质谱法同时测定谷物中12种真菌毒素[J].作物学报,2014,40(4):691-701.
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[14]DALL’ASTA C,SFORZA S,GALAVERNA G,et al.Simultaneous detection of type A and type B trichothecenes in cereals by liquid chromatography-electrospray ionization mass spectrometry using NaCl as cationization agent[J].Journal of Chromatography A,2004,1054(1):389-393.
[15]吴杰,刘太国,高利,等.小麦四种镰刀菌毒素的ELISA检测及毒素污染分析[J].植物保护学报,2009,36(2):106-112.DOI:10.7666/d.y1635190.
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[17]ROMAGNOLI B,FERRARI M,BERGAMINI C.Simultaneous determination of deoxynivalenol,zearalenone,T-2 and HT-2 toxins in breakfast cereals and baby food by high-performance liquid chromatography and tandem mass spectrometry[J].Journal of Mass Spectrometry,2010,45(9):1075-1080.DOI:10.1002/jms.1802.
[18]RADOVáZ,HOLADOVáK,HAJSLOVáJ.Comparison of two clean-up principles for determination of trichothecenes in grain extract[J].Journal of Chromatography A,1998,829(1):259-267.
[19]TANAKA T,YONEDA A,INOUE S,et al.Simultaneous determination of trichothecene mycotoxins and zenralenone in cereals by gas chromatography-mass spectrometry[J].Journal of Chromatography A,2000,882(1):23-28.
[20]VACLAVIKOVA M,MACMAHON S,ZHANG K,et al.Application of single immunoaffinity clean-up for simultaneous determination of regulated mycotoxins in cereals and nuts[J].Talanta,2013,117(22):345-351.
[21]LATTANZIO V M T,SOLFRIZZO M,POWERS S,et al.Simultaneous determination of aflatoxins,ochratoxin A and Fusarium toxins in maize by liquid chromatography/tandem mass spectrometry after multitoxin immunoaffinity cleanup[J].Rapid Communications in Mass Spectrometry,2007,21(20):3253-3261.
[22]郑翠梅,张艳,王松雪,等.液相色谱-质谱联用同时检测粮食中多种真菌毒素的应用进展[J].粮食科技与经济,2012,37(1):45-49.DOI:10.3969/j.issn.1007-1458.2012.01.022.
[23]FERREIRA I,FERNANDES J O,CUNHA S C.Optimization and validation of a method based in a QuEChERS procedure and gas chromatography-mass spectrometry for the determination of multimycotoxins in popcorn[J].Food Control,2012,27(1):188-193.DOI:10.1016/j.foodcont.2012.03.014.
[24]DIANA D,MAVUNGU J,MONBALIU S,et al.LC-MS/MS multianalyte method for mycotoxin determination in food supplements[J].Food Additives and Contaminants,2009,26(6):885-895.DOI:10.1080/02652030902774649.
[25]STECHER G,JARUKAMJORN K,ZABORSKI P,et al.Evaluation of extraction methods for the simultaneous analysis of simple and macrocyclic trichothecenes[J].Talanta,2007,73(2):251-257.DOI:10.1016/j.talanta.2007.03.028.
[26]MEI L Y,CAO B Y,YANG H,et al.Development of an immunoaffinity chromatography column forselective extraction of a new agonist phenylethylamine A from feed,meat and liver samples[J].Journal of Chromatography B,2014,945/946(2):178-184.
[27]RAZZAZI F E,BOM J,JARUKAMJORN K,et al.Simultaneous determination of major B-trichothecenes and the de-epoxymetabolite of deoxynivalenol in pig urine and maize using highperformance liquid chromatography-mass spectrometry[J].Journal of Chromatography B,2003,796(1):21-33.
[28]RAZZAZI F E,BOM J,LUF W.Determination of nivalenol and deoxynivalenol in wheat using liquid chromatography-mass spectrometry with negative ion atmospheric pressure chemical ionisation[J].Journal of Chromatography A,1999,854(1):45-55.DOI:10.1016/s0021-9673(99)00616-0.
[29]SCHOTHORST R C,JEKEL A A.Determination of trichothecenes in wheat by capillary gas chromatography with flame ionisation detection[J].Food Chemistry,2001,73(1):111-117.DOI:10.1016/s0308-8146(00)00321-6.
[30]静平,胥璐,马志勇,等.高效液相色谱法检测粮谷及其制品中六种A,B型单端孢霉烯族真菌毒素[J].食品安全质量检测学报,2012,3(2):106-111.
[31]鲍蕾,吴振兴,石媛媛,等.多功能柱净化-高效液相色谱法检测粮谷及其制品中的呕吐毒素[J].食品安全质量检测学报,2014,5(3):776-781.
[2]VISCONTI A,LATTANZIO V M,PASCALE M,et al.Analysis of T-2 and HT-2 toxins in cereal grains by immunoaffinity clean-up and liquid chromatography with fluorescence detection[J].Journal of Chromatography A,2005,1075(1/2):151-158.DOI:10.1016/j.chroma.2005.04.009.
[3]WILD C P,GONG Y Y.Mycotoxins and human disease:alargely ignored global health issue[J].Carcinogenesis,2010,31(1):71-82.DOI:10.1093/carcin/bgp264.
[4]PESTKA J J,ROTTER B A,PRELUSKY D B,et al.Toxicology of deoxynivalenol(vomitoxin)[J].Journal of Toxicology Environmental Health,1996,48(1):1-34.
[5]CHANEMOUGASOUNDHARAM A,FIONA M D.Trichothecene toxicity in eukaryotes:cellular and molecular mechanisms in plants and animals[J].Toxicology Letters,2013,217(2):149-158.
[6]李丽霞,尚书凤.T-2毒素的毒性及其作用机制与代谢研究进展[J].湖南农业科学,2015,54(21):5207-5210.DOI:10.14088/j.cnki.issn0439-8114.2015.21.002.
[7]梁颖.配合饲料中三种镰刀菌毒素检测方法的研究[D].咸阳:西北农林科技大学,2006.
[8]高爽,邓又天,张超,等.脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮及其联合染毒对小鼠肝脏功能及肝细胞凋亡的影响[J].中国兽医学报,2015,35(12):2021-2026.DOI:10.16303/j.cnki.1005-4545.2015.12.25.
[9]KOTAL F,HOLADOVáK,HAJSOVJ,et al.Determination of trichothecenes in cereals[J].Journal of Chromatography A,1999,830(1):219-225.
[10]EKE Z,KENDE A,TORKOS K.Simultaneous detection of A and B trichothecenes by gas chromatography with flame ionization or mass selective detection[J].Microchemical Journal,2004,78(2):211-216.
[11]SOLEIMANY F,JINAP S,RAHMANI A,et al.Simultaneous detection of 12 mycotoxins in cereals using RP-HPLC-PDA-FLD with PHRED and a post-column derivatization system[J].Food Additives and Contaminants,2011,28(4):494-501.
[12]孙娟,李为喜,张妍,等.用超高效液相色谱串联质谱法同时测定谷物中12种真菌毒素[J].作物学报,2014,40(4):691-701.
[13]ULRIKE B,INGA R,SUSANNE K,et al.Diagnosis of intoxications of piglets fed with Fusarium toxin-contaminated maize by the analysis of mycotoxin residues in serum,liquor and urine with LC-MS/MS[J].Archives of Animal Nutrition.2014,68(6):425-447.
[14]DALL’ASTA C,SFORZA S,GALAVERNA G,et al.Simultaneous detection of type A and type B trichothecenes in cereals by liquid chromatography-electrospray ionization mass spectrometry using NaCl as cationization agent[J].Journal of Chromatography A,2004,1054(1):389-393.
[15]吴杰,刘太国,高利,等.小麦四种镰刀菌毒素的ELISA检测及毒素污染分析[J].植物保护学报,2009,36(2):106-112.DOI:10.7666/d.y1635190.
[16]WANG Q,ZHAO H,XI C X,et al.Determination of chloramphenicol and zeranols in pig muscle by immunoaffinity column clean-up and LC-MS/MS analysis[J].Food Additives and Contaminants,2014,31(7):1177-1186.
[17]ROMAGNOLI B,FERRARI M,BERGAMINI C.Simultaneous determination of deoxynivalenol,zearalenone,T-2 and HT-2 toxins in breakfast cereals and baby food by high-performance liquid chromatography and tandem mass spectrometry[J].Journal of Mass Spectrometry,2010,45(9):1075-1080.DOI:10.1002/jms.1802.
[18]RADOVáZ,HOLADOVáK,HAJSLOVáJ.Comparison of two clean-up principles for determination of trichothecenes in grain extract[J].Journal of Chromatography A,1998,829(1):259-267.
[19]TANAKA T,YONEDA A,INOUE S,et al.Simultaneous determination of trichothecene mycotoxins and zenralenone in cereals by gas chromatography-mass spectrometry[J].Journal of Chromatography A,2000,882(1):23-28.
[20]VACLAVIKOVA M,MACMAHON S,ZHANG K,et al.Application of single immunoaffinity clean-up for simultaneous determination of regulated mycotoxins in cereals and nuts[J].Talanta,2013,117(22):345-351.
[21]LATTANZIO V M T,SOLFRIZZO M,POWERS S,et al.Simultaneous determination of aflatoxins,ochratoxin A and Fusarium toxins in maize by liquid chromatography/tandem mass spectrometry after multitoxin immunoaffinity cleanup[J].Rapid Communications in Mass Spectrometry,2007,21(20):3253-3261.
[22]郑翠梅,张艳,王松雪,等.液相色谱-质谱联用同时检测粮食中多种真菌毒素的应用进展[J].粮食科技与经济,2012,37(1):45-49.DOI:10.3969/j.issn.1007-1458.2012.01.022.
[23]FERREIRA I,FERNANDES J O,CUNHA S C.Optimization and validation of a method based in a QuEChERS procedure and gas chromatography-mass spectrometry for the determination of multimycotoxins in popcorn[J].Food Control,2012,27(1):188-193.DOI:10.1016/j.foodcont.2012.03.014.
[24]DIANA D,MAVUNGU J,MONBALIU S,et al.LC-MS/MS multianalyte method for mycotoxin determination in food supplements[J].Food Additives and Contaminants,2009,26(6):885-895.DOI:10.1080/02652030902774649.
[25]STECHER G,JARUKAMJORN K,ZABORSKI P,et al.Evaluation of extraction methods for the simultaneous analysis of simple and macrocyclic trichothecenes[J].Talanta,2007,73(2):251-257.DOI:10.1016/j.talanta.2007.03.028.
[26]MEI L Y,CAO B Y,YANG H,et al.Development of an immunoaffinity chromatography column forselective extraction of a new agonist phenylethylamine A from feed,meat and liver samples[J].Journal of Chromatography B,2014,945/946(2):178-184.
[27]RAZZAZI F E,BOM J,JARUKAMJORN K,et al.Simultaneous determination of major B-trichothecenes and the de-epoxymetabolite of deoxynivalenol in pig urine and maize using highperformance liquid chromatography-mass spectrometry[J].Journal of Chromatography B,2003,796(1):21-33.
[28]RAZZAZI F E,BOM J,LUF W.Determination of nivalenol and deoxynivalenol in wheat using liquid chromatography-mass spectrometry with negative ion atmospheric pressure chemical ionisation[J].Journal of Chromatography A,1999,854(1):45-55.DOI:10.1016/s0021-9673(99)00616-0.
[29]SCHOTHORST R C,JEKEL A A.Determination of trichothecenes in wheat by capillary gas chromatography with flame ionisation detection[J].Food Chemistry,2001,73(1):111-117.DOI:10.1016/s0308-8146(00)00321-6.
[30]静平,胥璐,马志勇,等.高效液相色谱法检测粮谷及其制品中六种A,B型单端孢霉烯族真菌毒素[J].食品安全质量检测学报,2012,3(2):106-111.
[31]鲍蕾,吴振兴,石媛媛,等.多功能柱净化-高效液相色谱法检测粮谷及其制品中的呕吐毒素[J].食品安全质量检测学报,2014,5(3):776-781.