酶联免疫法检测混合植物油中黄曲霉毒素B_1含量
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摘要
目的建立酶联免疫法测定混合植物油中黄曲霉毒素B_1含量的分析方法。方法采用直接竞争酶联免疫吸附法对样品进行测定。样品经甲醇溶液提取,提取的抗原及黄曲霉毒素B_1标记物与微孔板上的黄曲霉毒素B_1特异性单克隆抗体竞争结合,洗涤除去未结合杂质。将底物加入孵育,产生蓝色产物,加入终止液终止反应蓝色产物变成黄色产物,450nm处测量吸光度。结果本方法在2h内完成混合植物油中黄曲霉毒素B_1含量的测定。黄曲霉毒素B_1的加标浓度在1.0～20.0μg/kg之间时的回收率为76.5%～120.2%,方法定量限为1.00μg/kg。结论该方法快速、准确、灵敏,适合测定混合植物油中黄曲霉毒素B_1。
Objective To establish a method for the determination of aflatoxin B_1 in mixed vegetable oil by enzyme immunosorbent assay. Methods The samples were determined by direct competitive enzyme-linked immunosorbent assay. The samples were extracted by methanol solution, the extracted antigen was competitive bound with the aflatoxin B_1 marker and the specific monoclonal antibody against aflatoxin B_1 on the microporous plate, and the unbound impurities were removed by washing. When the substrate was incubated, producing blue product, and the blue product was changed into yellow product by adding the termination solution. The absorbance was measured at 450 nm. Results The determination of aflatoxin B_1 content in mixed vegetable oil was finished within 2 hours by this method. The recoveries of aflatoxin B_1 were 76.5%-120.2% when the concentration of aflatoxin B_1 was between 1.0-20.0 μ g/kg, and the limit of detection was 1.00 μg/kg. Conclusion This method is rapid, accurate, sensitive, which is suitable for the determination of aflatoxinB_1 in mixed vegetable oil.
Objective To establish a method for the determination of aflatoxin B_1 in mixed vegetable oil by enzyme immunosorbent assay. Methods The samples were determined by direct competitive enzyme-linked immunosorbent assay. The samples were extracted by methanol solution, the extracted antigen was competitive bound with the aflatoxin B_1 marker and the specific monoclonal antibody against aflatoxin B_1 on the microporous plate, and the unbound impurities were removed by washing. When the substrate was incubated, producing blue product, and the blue product was changed into yellow product by adding the termination solution. The absorbance was measured at 450 nm. Results The determination of aflatoxin B_1 content in mixed vegetable oil was finished within 2 hours by this method. The recoveries of aflatoxin B_1 were 76.5%-120.2% when the concentration of aflatoxin B_1 was between 1.0-20.0 μ g/kg, and the limit of detection was 1.00 μg/kg. Conclusion This method is rapid, accurate, sensitive, which is suitable for the determination of aflatoxinB_1 in mixed vegetable oil.
引文
[1]蔡飞,高微微,李红玲,等.中药上黄曲霉毒素的污染现状与防除技术[J].中国中药杂志,2010,(19):2503-2507.Cai F,Gao WW,Li HL,et al.Pollution status and control technology of aflatoxin in traditional Chinese medicine[J].China J Chin Mater Med,2010,(19):2503-2507.
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[3]谢体波,刘红,陆苇,等.间接竞争ELISA检测试剂盒测定粮油食品中黄曲霉毒素B1[J].食品安全质量检测学报,2015,6(7):2834-2839.Xie TB,Liu H,Lu W,et al.Determination of aflatoxin B1 in cereals and oils by indirect competition ELISA detection kit[J].J Food Saf Qual,2015,6(7):2834-2839.
[4]许菲菲,刘亚丽,田富饶.黄曲霉毒素检测方法研究[J].化学分析计量,2010,(5):93-94.Xu FF,Liu YL,Tian FR.Study on detection methods of aflatoxin[J].Chem Anal Meter,2010,(5):93-94.
[5]王君,刘秀梅.食品中真菌毒素危险性分析的方法及现状[J].中华预防医学杂志,2005,(6):430-433.Wang J,Liu XM.Current situation and analysis method of the risk of mycotoxins in food[J].Chin J Prev Med,2005,(6):430-433.
[6]GB 2716-2005中华人民共和国国家标准食用植物油卫生指标[S].GB 2716-2005 People's Republic of China national standard-Health index of edible vegetable oil[S].
[7]李岩松,周玉,谭建华.真菌毒素快速分析方法研究进展[J].中国卫生检验杂志,2006,(1):126-128.Li YS,Zhou Y,Tan JH.Advances in rapid analysis of mycotoxin[J].Chin J Health Lab Technol,2006,(1):126-128.
[8]Stroka J,Anklam E.Development of a simplified densitometer for the determination of aflatoxins by thin-layer chromatography[J].JChromatogr A,2000,904(2):263-268.
[9]柳其芳.酶联免疫吸附法和薄层色谱法联合分析黄曲霉毒素B1的研究[J].中国热带医学,2006,6(2):246-248.Liu QF.Study on the detection of aflatoxin B1 by ELISA and thin layer chromatography[J].China Trop Med,2006,6(2):246-248.
[10]彭晓俊,曾勋,庞普山,等.自制混合型固相萃取柱-高效液相色谱法同时测定食品中黄曲霉毒素B1、M1[J].分析测试学报,2013,32(8):958-962.Peng XJ,Zeng X,Pang PS,et al.Simultaneous determination of aflatoxin B1,M1 in food by high performance liquid chromatography with self-made mixed solid phase extraction column[J].J Instrum Anal,2013,32(8):958-962.
[11]Blesa J,Soriano JM,Molto JC.Determination of aflatoxins in peanuts by matrix solid-phase dispersion and liquid chromatography[J].JChromatogr A,2003,1011:49-54.
[12]Sheijooni-Fumani N,Hassan J,Yousefi R,et al.Determination of aflatoxin B1 in cereals by homogeneous liquid-liquid extraction coupled to high performance liquid chromatography-fluorescence detection[J].JSeparat Sci,2011,34:1333-1337.
[13]刘晓茂,李学民,王飞,等.超高压液相色谱-串联质谱法测定食用植物油中4种黄曲霉毒素B1、B2、G1、G2[J].食品安全质量检测学报,2012,3(5):513-518.Liu XM,Li XM,Wang F,et al.Determination of 4 aflatoxin B1,B2,G1,G2in edible vegetable oils by ultra high pressure liquid chromatography tandem mass spectrometry[J].J Food Saf Qual,2012,3(5):513-518.
[14]Kolosova AY,Shim WB,Yang ZY.Direct competitive ELISA based on a mondclonal antiboby for detection of aflatoxin B1 Stabilization of ELISAkit components and application to grain samples[J].Anal Bioanal Chem,2006,384(1):286-294.
[15]龚燕,赵春城,张东升,等.ELISA法检测五类食品中黄曲霉毒素B1前处理方法的改进研究[J].食品工业科技,2003,24(4):82-84.Gong Y,Zhao CC,Zhang DS,et al.Improvement of pretreatment of aflatoxin B1 in five kinds of foods by ELISA method[J].Sci Technol Food Ind,2003,24(4):82-84.
[16]美国Helica黄曲霉毒素B1检测试剂盒[EB/OL].[2010-10-18].http://blog.163.com/reagen5@126/blog/static/169929154201091842419495/.2010-10-18.Helica aflatoxin B1 detection kit[EB/OL].[2010-10-18].http://blog.163.com/reagen5@126/blog/static/169929154201091842419495/.2010-10-18.
[17]GB/T 27404-2008实验室质量控制规范食品理化检测[S].GB/T 27404-2008 Criterion on quality control of laboratories-Chemical testing of food[S].
[2]陈建民,张雪辉,杨美华,等.黄曲霉毒素检测方法研究进展[J].中国中药杂志,2005,30(24):1890-1894.Chen JM,Zhang XH,Yang MH,et al.Progress in the detection methods of aflatoxin[J].China J Chin Mater Med,2005,30(24):1890-1894.
[3]谢体波,刘红,陆苇,等.间接竞争ELISA检测试剂盒测定粮油食品中黄曲霉毒素B1[J].食品安全质量检测学报,2015,6(7):2834-2839.Xie TB,Liu H,Lu W,et al.Determination of aflatoxin B1 in cereals and oils by indirect competition ELISA detection kit[J].J Food Saf Qual,2015,6(7):2834-2839.
[4]许菲菲,刘亚丽,田富饶.黄曲霉毒素检测方法研究[J].化学分析计量,2010,(5):93-94.Xu FF,Liu YL,Tian FR.Study on detection methods of aflatoxin[J].Chem Anal Meter,2010,(5):93-94.
[5]王君,刘秀梅.食品中真菌毒素危险性分析的方法及现状[J].中华预防医学杂志,2005,(6):430-433.Wang J,Liu XM.Current situation and analysis method of the risk of mycotoxins in food[J].Chin J Prev Med,2005,(6):430-433.
[6]GB 2716-2005中华人民共和国国家标准食用植物油卫生指标[S].GB 2716-2005 People's Republic of China national standard-Health index of edible vegetable oil[S].
[7]李岩松,周玉,谭建华.真菌毒素快速分析方法研究进展[J].中国卫生检验杂志,2006,(1):126-128.Li YS,Zhou Y,Tan JH.Advances in rapid analysis of mycotoxin[J].Chin J Health Lab Technol,2006,(1):126-128.
[8]Stroka J,Anklam E.Development of a simplified densitometer for the determination of aflatoxins by thin-layer chromatography[J].JChromatogr A,2000,904(2):263-268.
[9]柳其芳.酶联免疫吸附法和薄层色谱法联合分析黄曲霉毒素B1的研究[J].中国热带医学,2006,6(2):246-248.Liu QF.Study on the detection of aflatoxin B1 by ELISA and thin layer chromatography[J].China Trop Med,2006,6(2):246-248.
[10]彭晓俊,曾勋,庞普山,等.自制混合型固相萃取柱-高效液相色谱法同时测定食品中黄曲霉毒素B1、M1[J].分析测试学报,2013,32(8):958-962.Peng XJ,Zeng X,Pang PS,et al.Simultaneous determination of aflatoxin B1,M1 in food by high performance liquid chromatography with self-made mixed solid phase extraction column[J].J Instrum Anal,2013,32(8):958-962.
[11]Blesa J,Soriano JM,Molto JC.Determination of aflatoxins in peanuts by matrix solid-phase dispersion and liquid chromatography[J].JChromatogr A,2003,1011:49-54.
[12]Sheijooni-Fumani N,Hassan J,Yousefi R,et al.Determination of aflatoxin B1 in cereals by homogeneous liquid-liquid extraction coupled to high performance liquid chromatography-fluorescence detection[J].JSeparat Sci,2011,34:1333-1337.
[13]刘晓茂,李学民,王飞,等.超高压液相色谱-串联质谱法测定食用植物油中4种黄曲霉毒素B1、B2、G1、G2[J].食品安全质量检测学报,2012,3(5):513-518.Liu XM,Li XM,Wang F,et al.Determination of 4 aflatoxin B1,B2,G1,G2in edible vegetable oils by ultra high pressure liquid chromatography tandem mass spectrometry[J].J Food Saf Qual,2012,3(5):513-518.
[14]Kolosova AY,Shim WB,Yang ZY.Direct competitive ELISA based on a mondclonal antiboby for detection of aflatoxin B1 Stabilization of ELISAkit components and application to grain samples[J].Anal Bioanal Chem,2006,384(1):286-294.
[15]龚燕,赵春城,张东升,等.ELISA法检测五类食品中黄曲霉毒素B1前处理方法的改进研究[J].食品工业科技,2003,24(4):82-84.Gong Y,Zhao CC,Zhang DS,et al.Improvement of pretreatment of aflatoxin B1 in five kinds of foods by ELISA method[J].Sci Technol Food Ind,2003,24(4):82-84.
[16]美国Helica黄曲霉毒素B1检测试剂盒[EB/OL].[2010-10-18].http://blog.163.com/reagen5@126/blog/static/169929154201091842419495/.2010-10-18.Helica aflatoxin B1 detection kit[EB/OL].[2010-10-18].http://blog.163.com/reagen5@126/blog/static/169929154201091842419495/.2010-10-18.
[17]GB/T 27404-2008实验室质量控制规范食品理化检测[S].GB/T 27404-2008 Criterion on quality control of laboratories-Chemical testing of food[S].