食用油中邻苯二甲酸二丁酯高灵敏检测技术建立与应用  

Establishment and application of a highly sensitive detection technique for di-butyl phthalate in edible oils

作　　者：向成艳 刘品品 唐晓倩[1,3,4,5,6] 张奇 李培武[1,3,4,5,6,7] XIANG Cheng-yan;LIU Pin-pin;TANG Xiao-qian;ZHANG Qi;LI Pei-wu(Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences,Wuhan 430062,China;Wuhan Polytechnic University,Wuhan 430023,China;Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture and Rural Affairs,Wuhan 430062,China;Key Laboratory of Detection for Mycotoxins,Ministry of Agriculture and Rural Affairs,Wuhan 430062,China;Laboratory of Risk Assessment for Oilseeds Products,Ministry of Agriculture and Rural Affairs,Wuhan 430062,China;Quality Inspection and Test Center for Oilseeds Products,Ministry of Agriculture and Rural Affairs,Wuhan 430062,China;Hubei Hongshan Laboratory,Wuhan 430070,China)

机构地区：[1]中国农业科学院油料作物研究所,湖北武汉430062 [2]武汉轻工大学,湖北武汉430023 [3]农业农村部油料作物生物学与遗传育种重点实验室,湖北武汉430062 [4]农业农村部生物毒素检测重点实验室,湖北武汉430062 [5]农业农村部油料产品质量安全风险评估实验室,湖北武汉430062 [6]农业农村部油料及制品质量监督检验测试中心,湖北武汉430062 [7]湖北省洪山实验室,湖北武汉430070

出　　处：《中国油料作物学报》2024年第5期1140-1147,共8页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金青年基金(32102089);中国农业科学院科技创新工程重大科研任务(CAAS-XTCX2019024)。

摘　　要：食用油在生产、加工、储存各环节中,易受邻苯二甲酸二丁酯(di-butyl phthalate,DBP)污染。本文以单克隆抗体为识别元件,建立并优化了食用油DBP的高灵敏酶联免疫吸附检测方法(Enzyme-linked immunosorbent assay,ELISA)。结果显示,在抗原包被浓度为1μg/mL,4%脱脂乳粉作为封闭剂,磷酸缓冲液pH 7.4,离子浓度为20mmol/L,甲醇浓度为40%条件下,该ELISA方法的灵敏度(IC_(50))为16.29 ng/mL,检测范围为0.33~94.45 ng/mL(IC_(90)~IC_(10)),LOD(IC_(90))为0.33 ng/mL。通过食用油加标试验建立基质标准曲线,灵敏度(IC_(50))为92.57 ng/mL,检测范围为2.44~606.12 ng/mL(IC_(90)~IC_(10)),LOD(IC_(90))为2.44 ng/mL。在食用油样品中分别添加浓度为40 ng/mL、120 ng/mL、200 ng/mL的DBP标准品,回收率为71.34%~97.23%,变异系数为1.05%~6.38%,采用建立的DBP-ELISA检测方法分别对大豆油、玉米油等实际样品进行检测,结果与国标检测方法相关性达到99.17%,表明建立的ELISA方法可用于食用油实际样品中DBP含量的测定。Edible oils are susceptible to contamination by di-butyl phthalate(DBP)during various stages of production,processing and storage.In this paper,we have developed and optimized a highly sensitive ELISA meth-od for the determination of di-butyl phthalate in edible oil using monoclonal antibodies as the recognition element.The results demostrated that the sensitivity(IC_(50))of the ELISA method was 16.29 ng/mL,and the detection range was 0.33~94.45 ng/mL(IC_(90)~IC_(10))at an antigen encapsulation concentration of 1μg/mL,with 4%skimmed milk powder as the blocking agent,and the sample was used at pH 7.4 with an ionic concentration of 20 mmol/L in 40%methanol-PBS.The sensitivity(IC_(50))was 92.57 ng/mL,the detection range was 2.44-606.12 ng/mL(IC_(90)-IC_(10)),and the LOD(IC_(90))was 2.44 ng/mL.The matrix standard curve was established by the edible oil spiking test,and the sensitivity(IC_(50))was 92.57 ng/mL,the detection range was 2.44-606.12 ng/mL(IC_(90)-IC_(10)),and the LOD(IC_(90))was 2.44 ng/mL.The edible oil samples were spiked with DBP standards of 40 ng/mL,120 ng/mL and 200 ng/mL,re-spectively.The recoveries were 71.34%-97.23%,and the coefficients of variation were 1.05%-6.38%.The estab-lished DBP ELISA method was used to test the actual samples of soybean oil and corn oil,and the correlation be-tween the results and the national standard test method reached 99.17%,indicating that the method can be used for the determination of DBP content in the actual samples of edible oil.

关 键 词：食用油 邻苯二甲酸二丁酯 酶联免疫吸附 灵敏度 

分 类 号：TS207.3[轻工技术与工程—食品科学]