多组分免疫亲和柱净化/超高效液相色谱-串联质谱检测水产饲料中的黄曲霉毒素
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摘要
建立了多组分免疫亲和柱净化/超高效液相色谱-串联质谱(UPLC-MS/MS)测定水产饲料中黄曲霉毒素AFB_1、AFB_2、AFG_1及AFG_2含量的方法。饲料样品用80%乙腈水超声提取后,经多组分免疫亲和柱净化,采用UPLC-MS/MS测定,外标法定量。以0.1%甲酸-乙腈为流动相,梯度洗脱分离,电喷雾正离子多反应监测模式检测。结果表明,AFB_1、AFG_1和AFB_2、AFG_2分别在2.25～22.5 ng/mL和0.75～7.5 ng/mL质量浓度范围内呈良好线性,相关系数大于0.997;AFB_1、AFG_1的定量下限均为0.7μg/kg,AFB_2、AFG_2的定量下限均为0.2μg/kg。AFB_1、AFG_1在1.5μg/kg,AFB_2、AFG_2在0.5μg/kg加标水平下的回收率为78.7%～85.5%,日内相对标准偏差(RSD)为6.0%～6.5%,日间RSD为6.6%～7.0%。该法操作简单,耗时少,重复性好,灵敏度高,适用于水产饲料中黄曲霉毒素的测定。
An ultra performance liquid chromatography-tandem mass spectrometric(UPLC-MS/MS) method was established for the determination of aflatoxins(AFB_1,AFB_2,AFG_1 and AFG_2)in aquatic feeds based on multi-component immunoaffinity column purification.The samples were ultrasonically extracted with 80%acetonitrile aqueous solution,then purified with a multi-component immunoaffinity column,finally analyzed by UPLC-MS/MS with electrospray ionization in positive ion mode(ESI~+)under multiple reaction monitoring(MRM)mode and quantified by the external standard method.The separation was performed on an Acquity UPLC BEH C_(18) column by gradient elution with 0.1%formic acid-acetonitrile as mobile phases.Good linear responses were obtained in the concentration ranges of 2.25-22.5 ng/mL for AFB_1 and AFG_1,and 0.75-7.5 ng/mL for AFB_2 and AFG_2,with their correlation coefficients larger than 0.997,respectively.The limits of quantitation(LOQs) were 0.7 μg/kg for AFB_1 and AFG_1,and 0.2 μg/kg for AFB_2 and AFG_2,respectively.The average recoveries for AFB_1 and AFG_1 at a spiked level of 1.5 μg/kg,and AFB_2 and AFG_2 at a spiked level of 0.5 μg/kg ranged from 78.7%to 85.5%,with intra-day and inter-day relative standard deviations of 6.0%-6.5%and 6.6%-7.0%,respectively.The method was simple,rapid,sensitive and reliable,and was suitable for the determination of aflatoxins residues in aquatic feed samples.
An ultra performance liquid chromatography-tandem mass spectrometric(UPLC-MS/MS) method was established for the determination of aflatoxins(AFB_1,AFB_2,AFG_1 and AFG_2)in aquatic feeds based on multi-component immunoaffinity column purification.The samples were ultrasonically extracted with 80%acetonitrile aqueous solution,then purified with a multi-component immunoaffinity column,finally analyzed by UPLC-MS/MS with electrospray ionization in positive ion mode(ESI~+)under multiple reaction monitoring(MRM)mode and quantified by the external standard method.The separation was performed on an Acquity UPLC BEH C_(18) column by gradient elution with 0.1%formic acid-acetonitrile as mobile phases.Good linear responses were obtained in the concentration ranges of 2.25-22.5 ng/mL for AFB_1 and AFG_1,and 0.75-7.5 ng/mL for AFB_2 and AFG_2,with their correlation coefficients larger than 0.997,respectively.The limits of quantitation(LOQs) were 0.7 μg/kg for AFB_1 and AFG_1,and 0.2 μg/kg for AFB_2 and AFG_2,respectively.The average recoveries for AFB_1 and AFG_1 at a spiked level of 1.5 μg/kg,and AFB_2 and AFG_2 at a spiked level of 0.5 μg/kg ranged from 78.7%to 85.5%,with intra-day and inter-day relative standard deviations of 6.0%-6.5%and 6.6%-7.0%,respectively.The method was simple,rapid,sensitive and reliable,and was suitable for the determination of aflatoxins residues in aquatic feed samples.
引文
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[2] Nyamete F A.Potential of Lactic Acid Fermentation in Reducing Aflatoxin B1 and Fumonisin B1 in Tanzanian Maize-Based Complementary Gruel.Michigan:Michigan State University,2013.
[3] Medina-Martínez M S,Martínez A J.J.Agric.Food Chem.,2000,48(7):2833-2836.
[4] Mwalwayo D S,Thole B.Toxicol.Reports,2016,3:173-179.
[5] Akiyama H,Goda Y,Tanaka T.J.Chromatogr.A,2001,932(1/2):153-157.
[6] Kamkar A,Aliabadi F S,Khaksar R.J.Veter.Res.,2008,63(2):7-12.
[7] Wu Y N.Present Knowledge in Food Safety.Beijing:Chemical Industry Press(吴永宁.现代食品安全科学.北京:化学工业出版社),2003:279.
[8] Steyn P S.Toxicol.Lett.,1995,82/83:843-851.
[9] Kong Q,Lin H,Guan B.Food Sci.(孔青,林洪,管斌.食品科学),2013,34(15):324-328.
[10] Li S G,Chen H,Li X M,Ren Y Y.Science and Technology of Cereals,Oils and Foods(李书国,陈辉,李雪梅,任媛媛.粮油食品科技),2009,17(2):62-65.
[11] Jiang J Y,Yang X W.Jiangxi Feed(蒋建云,杨学文.江西饲料),2006,(6):18-19.
[12] Jia T.Feed China(贾涛.饲料广角),2013,(13):34-35.
[13] Xing L,Wang R H.Feed and Husbandry(邢磊,王仁华.饲料与畜牧),2015,(7):55-56.
[14] Feng T,Qiao K Y,Zhou J.Mod.Sci.Instrum.(冯婷,乔坤云,周俊.现代科学仪器),2005,16(1):115-116.
[15] Chen X,Rao Z H,Ma D X,Wang X,Wang L.Chin.J.Animal Sci.(陈新,饶正华,马东霞,王雄,王莲.中国畜牧杂志),2007,43(24):50-54.
[16] Wang J R,Lin H,Liu C Y.Science and Technology of Cereals,Oils and Foods(王江蓉,林华,刘纯友.粮油食品科技),2012,20(2):34-36.
[17] Li L,Li R,Xie G,Ye J,Wang S X.J.Instrum.Anal.(李丽,黎睿,谢刚,叶金,王松雪.分析测试学报),2017,36(6):800-804.
[18] Song Y,Chen Y,Bai X.Pract.Prevent.Med.(宋月,陈颖,白欣.实用预防医学),2016,23(7):882-884.
[19] Zhu W X,Yang J Z,Yuan P,Wang C J,Wei W.Phys.Test.Chem.Anal.:Chem.Anal.(祝伟霞,杨冀州,袁萍,王彩娟,魏蔚.理化检验-化学分册),2013,49(8):981-984.
[20] Wang Q,Hang X Y,Song X.Chin.J.Health Lab.Technol.(王芹,杭学宇,宋鑫.中国卫生检验杂志),2016,(4):482-485.
[21] Liu F,Ren A S,Ge P,Ding N F,Naren Q Q G.J.Instrum.Anal.(刘飞,任安书,葛萍,丁年芳,娜仁·琪琪格.分析测试学报),2018,37(6):696-701.
[22] Li P P,Zhang X J,Mei G M,Yan Z Y,Yu L.J.Zhejiang Univ.:Sci.Ed.(李佩佩,张小军,梅光明,严忠雍,喻亮.浙江大学学报:理学版),2015,42(3):334-338.
[23] Li P P,Zhang X J,Yan Z Y,Chen S,Lun L L.Food Sci.(李佩佩,张小军,严忠雍,陈思,伦丽丽.食品科学),2016,37(24):257-261.
[24] Pan Y F,Zheng Y L,Fang C J,Chen J Y.Fujian Anal.Test.(潘迎芬,郑育莉,方成俊,陈俊玉.福建分析测试),2013,22(4):12-17.
[25] Ma Z F.J.Food Safe.Qual.(马占峰.食品安全质量检测学报),2012,3(3):209-211.
[26] Zheng R X,Han D,Zhu H M,Wang Y N,Zheng W B.Cereals and Oils Processing:Electron.Ver.(郑睿行,韩丹,祝华明,王怡念,郑微波.粮油加工:电子版),2014,(9):51-53.
[27] Jiang Z X,Liu Z W,Cao X,Wang Z L,Li Z J.Feed Ind.(姜兆兴,刘振伟,曹旭,王智亮,李智瑾.饲料工业),2007,28(20):52-54.
[28] Shang Q Q,Xiong P W,Liang Q,Rao Z H,Wang X,Guo Q,Yu D Y.Feed Res.(尚沁沁,熊平文,梁权,饶正华,王雄,果旗,余东游.饲料研究),2015,(22):55-60.
[29] Shi Y H,Xu Z R,Feng J L,Li J S.Chin.J.Animal Sci.(史莹华,许梓荣,冯建蕾,李家胜.中国畜牧杂志),2005,41(11):40-42.
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