【作者】 胡巍； 高金燕； 陈红兵； 李欣；

【Author】 Hu Wei;Gao Jinyan;Chen Hongbing;Li Xin;State Key Laboratory of Food Science and Resources,Nanchang University;College of Food Science &Technology,Nanchang University;Jiangxi Province Key Laboratory of Food Allergy;Sino German Joint Research Institute,Nanchang University;

【机构】 南昌大学食品科学与资源挖掘全国重点实验室； 南昌大学食品学院； 江西省食物过敏重点实验室； 南昌大学中德联合研究院；

【摘要】 有效的样品前处理可以提升检测的准确性。脂肪作为食品的常见成分,会与牛乳过敏原蛋白之间易发生相互作用从而影响过敏原蛋白的提取效果和检测准确性。本文首次建立了食物过敏原ELISA检测的预处理方法。以含有牛乳过敏原的巧克力为原料,分别测定了正己烷、异丙醇、乙酸乙酯和脂肪酶脱脂处理后复杂基质的脱脂率和蛋白损失率,比较了样品中蛋白结构、粒度、脂肪分布和微观结构的变化,同时利用间接ELISA评估样品中主要牛乳过敏原的溶出率。研究结果表明,正己烷的脱脂率最高且蛋白损失率最低,脱脂会改变蛋白质空间结构,整体趋势由紧凑变为松散。样品粒径在脱脂后略微降低,但异丙醇处理会使蛋白聚集成较大的颗粒。正己烷和乙酸乙酯脱脂后样品的脂肪分布更为均一且液滴变小,样品表面形貌由致密光滑块状变为疏松多孔结构,而异丙醇和脂肪酶组仍然存在部分结块。间接ELISA结果表明,食物过敏原在乙酸乙酯组中的溶出率提升,在正己烷组中的溶出浓度最高。综上所述,正己烷和乙酸乙酯脱脂处理可以明显改善脂肪复杂基质中过敏原的提取效果。本文建立的预处理方法在食物过敏原检测中具有良好的回收率、稳定性和准确性。更多还原

【Abstract】 Effective sample pretreatment can improve the accuracy of detection.As a common component of food,fat will easily interact with milk allergen protein,which will affect the extraction effect and detection accuracy of allergen protein.In this paper,a pretreatment method for the detection of food allergens by ELISA was established for the first time.The degreasing rates and protein loss rates of complex substrates after degreasing of n-hexane,isopropyl alcohol,ethyl acetate and lipase were determined,respectively.The changes in protein structure,particle size,fat distribution and microstructure of the samples were compared,and the dissolution rates of major milk allergens in the samples were evaluated by indirect ELISA.The results showed that the degreasing rate of n-hexane was the highest and the protein loss rate was the lowest.Degreasing changed the spatial structure of the protein from compact to loose.The particle size of the sample decreased slightly after degreasing,but the isopropyl alcohol treatment caused the protein to aggregate into larger particles.After the degreasing of nhexane and ethyl acetate,the fat distribution of the sample became more uniform and the droplet became smaller.The surface morphology of the sample changed from dense smooth block to loose porous structure,while some caking still existed in the isopropyl alcohol and lipase groups.The results of indirect ELISA showed that the dissolution rate of food allergens increased in ethyl acetate group,and the dissolution concentration was the highest in n-hexane group.In conclusion,the degreasing of n-hexane and ethyl acetate can obviously improve the extraction of allergens from the complex matrix of fat.The pretreatment method established in this paper showed good recovery,stability and accuracy in the detection of food allergens.更多还原