[目的]本研究使用酶联免疫法（ELISA法）评估了一级水、无水甲醇、二氯甲烷3 种不同提取剂在检测芝士粉中黄曲霉毒素M1含量方面的效果。[方法]通过对比一级水、无水甲醇、二氯甲烷3 种不同提取剂提取效果的相对标准偏差,比较平均回收率。[结果]用一级水提取时,相对标准偏差2.56%,平均回收率35.30%~51.12%;用无水甲醇提取时,相对标准偏差4.20%,平均回收率81.32%~90.18%;用二氯甲烷提取时,样品组织状态异常,氮流蒸发后残留物呈油状,无法进行后续检测。[结论]以无水甲醇作为提取剂时,ELISA法的提取准确性较高,更适用于食品检测领域。
[Objective] Enzyme-linked immunoassay（ELISA）was used to evaluate the effects of three different extractants of primary water,anhydrous methanol and dichloromethane on the determination of aflatoxin M1 in cheese powder. [Method] The average recovery rate was compared by comparing the relative standard deviation of extraction effects of primary water,anhydrous methanol and dichloromethane. [Result] When extracted with primary water,the relative standard deviation was 2.56% and the average recovery was 35.30%~51.12%. When extracted with anhydrous methanol,the relative standard deviation was 4.20%,and the average recovery was 81.32%~90.18%. When extracted with dichloromethane,the tissue status of the sample was abnormal,and the residue after nitrogen evaporation was oily,which could not be further determinated. [Conclusion] When extracted with anhydrous methanol. ELISA method had higher extraction accuracy and is more suitable for food determination.
[1] 华律.世界卫生组织国际癌症研究机构致癌物清单[EB/OL].https://m.66law.cn/contractmodel/5589955.aspx,2025-01-09.
[2] Umaya S R,Vijayalakshmi Y C,Sejian V.Exploration of plant products and phytochemicals against aflatoxin toxicity in broiler chicken production:Present status[J].Toxicon,2021,200:55-68.
[3] Ostry V,Malir F,Toman J,et al.Mycotoxins as human carcinogens—the IARC monographs classification[J].Mycotoxin Research,2017,33(1):65-73.
[4] Du B B,Su X O,Yang K H,et al.Antibody-free colorimetric detection of total aflatoxins in rice based on a simple two-step chromogenic reaction[J].Analytical Chemistry,2016,88(7):3775-3780.
[5] 卢晓霞,谢海军,宋崴,等.黄曲霉毒素M1酶联免疫吸附检测方法的建立[J] .食品工程,2017(2):58-60.
[6] 中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会. GB/T 33411—2016 酶联免疫分析试剂盒通则[S].
[7] 中华人民共和国国家质量监督检验检疫总局. SN/T2775—2011,商品化食品检测试剂盒评价方法[S].
[8] 中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会. GB/T 27403—2008,实验室质量控制规范食品分子生物学检测[S].
[9] 刘冬儿. 酶联免疫吸附分析法测定食品中的黄曲霉毒素B1[J].食品工业科技,2002,23(10):79-80.
[10] Shundo L,Sabino M.Aflatoxin M1 in milk by immunoaffinity column cleanup with TLC/HPLC determination[J].Brazilian Journal of Microbiology,2006,37(2):164-167.
[11] Chen M,Liu X,Yang S,et al.HPLC-MS/MS method for the simultaneous determination of aflatoxins in blood:Toxicokinetics of aflatoxin B1 and aflatoxin M1 in rats[J].Journal of Analytical Science and Technology,2022,13(1):27-11.
[12] Shuib N S,Makahleh A,Salhimi S M,et al.Determination of aflatoxin M1 in milk and dairy products using high performance liquid chromatography-fluorescence with post column photochemical derivatization[J].Journal of Chromatography A,2017,1510:51-56.
[13] 乔宽,王路阳,王艳飞,等.酶联免疫吸附和胶体金技术对黄曲霉毒素M1检测的结合验证[J].中国乳业,2024(1):69-72,77.
