黄曲霉毒素B_1时间分辨荧光定量检测体系适用性评价
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摘要
目的评价黄曲霉毒素B_1时间分辨荧光定量检测体系(包括黄曲霉毒素B_1时间分辨荧光免疫层析检测卡和荧光定量检测仪)的适用性。方法用时间分辨荧光定量检测体系测定20个阴性样本中黄曲霉毒素B_1的含量,确定该检测体系的检出限(limit of detection,LOD)和定量限(limit of quantification,LOQ);用2台荧光检测仪对6组含不同浓度黄曲霉毒素B_1的大米和玉米阳性样本进行检测,确定方法的准确性和台间差;重复6次检测含中等浓度黄曲霉毒素B_1的阳性样本和连续12 h检测国标检测限浓度的阳性标准溶液,确定方法的重复性和稳定性。结果该检测体系的LOD和LOQ分别为0.7和2.1μg/kg;该检测体系的检测值与大型仪器定值结果无显著性差异,2台荧光检测仪的测值之间无显著性差异;中浓度阳性样本和连续12 h测定检测限浓度的标准溶液的标准偏差未超过国家标准规定要求,检测结果的稳定性和重复性满足国家标准要求。结论该时间分辩荧光定量检测体系能够准确检测玉米和大米样本中黄曲霉毒素B_1含量。
Objective To evaluate the applicability of time-resolved fluoro-immunoassay quantitative detection system for aflatoxins B_1(including time-resolved fluorescence quantitative test strips and fluorescence quantitative detector). Methods The content of aflatoxin B_1 in 20 negative samples were detected by time-resolved fluoro-immunoassay quantitative detection system, and the limit of detection(LOD) and limit of quantitation(LOQ) of the detection system were determinated. Six sets of rice and corn positive samples containing different concentrations of aflatoxin B_1 were tested with 2 fluorescence detectors to determine the accuracy of the method and the difference between the stations. A positive sample containing a medium concentration of aflatoxin B_1 and a positive standard solution for detecting the concentration of the national standard detection limit were repeated 6 times to determine the repeatability and stability of the method. Results The LOD and LOQ were 0.7 and 2.1 μg/kg, respectively. There was no significant difference between the measured value of the test system and the value of the large-scale instrument, and there was no significant difference between the measured values of the 2 fluorescence detectors. Conclusion The time-resolved fluoro-immunoassay detection system can accurately detect the content of aflatoxins B_1 in rice and corn samples.
Objective To evaluate the applicability of time-resolved fluoro-immunoassay quantitative detection system for aflatoxins B_1(including time-resolved fluorescence quantitative test strips and fluorescence quantitative detector). Methods The content of aflatoxin B_1 in 20 negative samples were detected by time-resolved fluoro-immunoassay quantitative detection system, and the limit of detection(LOD) and limit of quantitation(LOQ) of the detection system were determinated. Six sets of rice and corn positive samples containing different concentrations of aflatoxin B_1 were tested with 2 fluorescence detectors to determine the accuracy of the method and the difference between the stations. A positive sample containing a medium concentration of aflatoxin B_1 and a positive standard solution for detecting the concentration of the national standard detection limit were repeated 6 times to determine the repeatability and stability of the method. Results The LOD and LOQ were 0.7 and 2.1 μg/kg, respectively. There was no significant difference between the measured value of the test system and the value of the large-scale instrument, and there was no significant difference between the measured values of the 2 fluorescence detectors. Conclusion The time-resolved fluoro-immunoassay detection system can accurately detect the content of aflatoxins B_1 in rice and corn samples.
引文
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[11]Alinorm 93/12.FAO/WHO Standards Programme[S].
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[13]Sun DD,Gu X,Li J,et al.Quality evaluation of five commercial enzyme linked-immunosorbent assay kits for detecting aflatoxin B1 in feedstuffs[J].Asian Austral J Anim Sic,2015,28(5):691-696.
[14]Amirkhizi B,Arefhosseini SR,Ansarin M,et al.Aflatoxin B1 in eggs and chicken livers by dispersive liquid-liquid microextraction and HPLC[J].Food Addit Contam B,2015,8(4):245-249.
[15]张兆威,李培武,张奇,等.农产品中黄曲霉毒素的时间分辨荧光免疫层析快速检测技术研究[J].中国农业科学,2014,47(18):3668-3674.Zhang ZW,LI PW,Zhang Q,et al.Study on time-resolved flurescence immunochromatography for aflatoxin determination in agricultural product[J].Sci Agric Sin,2014,47(18):3668-3674.
[16]李静,李培武,张奇,等.时间分辨荧光免疫层析试纸条在油料饼粕黄曲霉毒素B1检测中的应用[J].中国油料作物学报,2014,36(2):256-262.Li J,Li PW,Zhang Q,et al.Investigation of time-resolved fluoroimmunochromatographic assay strip for quantitative determination of aflatoxin B1 in oilseed meals[J].Chin J Oil Crop Sci,2014,36(2):256-262.
[17]LS/T 6402-2017粮油检验设备和方法标准适用性验证及结果评价一般原则[S].LS/T 6402-2017 Grain and oil inspection-General principles for verification of applicability of equipment and method standards and evaluation of results[S].
[18]GB/T 4889-2008数据的统计处理和解释正态分布均值和方差的估计与检验[S].GB/T 4889-2008 Statistical processing and interpretation of data-The estimation and test of the mean and variance of normal distribution[S].
[2]Kucukcakan B,Hayrulai-Musliu Z.Challenging role of dietary aflatoxin B1 exposure and hepatitis B infection on risk of hepatocellular carcinoma[J].Open Access Maced J Med Sci,2015,3(2):363-369.
[3]孙桂菊.肝癌高发地区人群黄曲霉毒素暴露水平的评估[D].厦门:复旦大学,2001.Sun GJ.Assessment of aflatoxin exposure in populations with high incidence of liver cancer[D].Xiamen:Fudan University,2001.
[4]Azab RM,Tawakkol WM,Abdel-Rahman M.et al.Detection and estimation of aflatoxin B1 in feeds and its biodegradation by bacteria and fungi[J].Egyp J Nat Toxin.2005,(2):39-56.
[5]Coulombe RA.Biological action of mycotoxins[J].J Dairy Sci,1993,(76):880-891.
[6]GB 2761-2017食品安全国家标准食品中真菌毒素限量[S].GB 2761-2017 National food safety standard-Limit of mycotoxins in food[S].
[7]Park DL,Njapau H.Contamination issues and padding[J].J Am Oil Chem Sot,1989,(66):1402-1405.
[8]Park DL,Liang B.Perspectives on aflatoxin control for human food and animal feed[J].Trend Food Sci Technol,1993,(4):334-342.
[9]EU 2015/863.Amending the annexes to Council Directive 74/63/EEC on undesirable substances and products in animal nutrition[S].
[10]Alinorm 91/29.FAO/WHO Standards Programme[S].
[11]Alinorm 93/12.FAO/WHO Standards Programme[S].
[12]Michele H,Juliane EW,Beatriz IN,et al.Photometric procedure for quantitative analysis of aflatoxin B1 in peanuts by thin-layer chromatography using charge coupled device detector[J].Química Nov,2008,31(3):514-517.
[13]Sun DD,Gu X,Li J,et al.Quality evaluation of five commercial enzyme linked-immunosorbent assay kits for detecting aflatoxin B1 in feedstuffs[J].Asian Austral J Anim Sic,2015,28(5):691-696.
[14]Amirkhizi B,Arefhosseini SR,Ansarin M,et al.Aflatoxin B1 in eggs and chicken livers by dispersive liquid-liquid microextraction and HPLC[J].Food Addit Contam B,2015,8(4):245-249.
[15]张兆威,李培武,张奇,等.农产品中黄曲霉毒素的时间分辨荧光免疫层析快速检测技术研究[J].中国农业科学,2014,47(18):3668-3674.Zhang ZW,LI PW,Zhang Q,et al.Study on time-resolved flurescence immunochromatography for aflatoxin determination in agricultural product[J].Sci Agric Sin,2014,47(18):3668-3674.
[16]李静,李培武,张奇,等.时间分辨荧光免疫层析试纸条在油料饼粕黄曲霉毒素B1检测中的应用[J].中国油料作物学报,2014,36(2):256-262.Li J,Li PW,Zhang Q,et al.Investigation of time-resolved fluoroimmunochromatographic assay strip for quantitative determination of aflatoxin B1 in oilseed meals[J].Chin J Oil Crop Sci,2014,36(2):256-262.
[17]LS/T 6402-2017粮油检验设备和方法标准适用性验证及结果评价一般原则[S].LS/T 6402-2017 Grain and oil inspection-General principles for verification of applicability of equipment and method standards and evaluation of results[S].
[18]GB/T 4889-2008数据的统计处理和解释正态分布均值和方差的估计与检验[S].GB/T 4889-2008 Statistical processing and interpretation of data-The estimation and test of the mean and variance of normal distribution[S].