橘霉素免疫牛蛙肌肉蛋白柱的制备及其吸附橘霉素作用分析  

Preparation and Adsorption Performance of Immunoaffinity Column Based on Muscle Antibody from Bullfrog(Rana catesbeiana) for Detection of Citrinin

作　　者：王向阳[1] 顾双[1] 杨玲[1] WANG Xiangyang;GU Shuang;YANG Ling(School of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018,China)

机构地区：[1]浙江工商大学食品与生物工程学院,浙江杭州310018

出　　处：《食品科学》2018年第22期159-165,共7页Food Science

基　　金：浙江省自然科学基金项目(LY15C200005)

摘　　要：用橘霉素人工抗原-佐剂免疫牛蛙,研究牛蛙肌肉蛋白质对橘霉素结合能力的大小。将能与橘霉素结合的蛋白质用海藻酸钠制粒。取6.4 mg橘霉素人工抗原免疫牛蛙肌肉蛋白质,使用海藻酸钠质量浓度4 g/100 mL、氯化钙质量浓度5 g/100 mL、戊二醛交联体积分数0.2%、交联时间2 h,制备海藻酸钙包埋蛋白质凝胶微球。取海藻酸钙凝胶微球2 g装填成固相小柱,以2 mL红曲米橘霉素提取液过柱,以20 mL超纯水洗脱杂质,与市售免疫亲和柱色谱图比较,均没有杂质干扰峰。以橘霉素标准液作为上样液,以70%乙腈-三氟乙酸作为橘霉素洗脱液,洗脱剂量为5 mL,回收率为86.7%,柱容量为0.101μg/g。采用自制的牛蛙免疫蛋白柱和市售免疫亲和柱测得红曲米中橘霉素含量分别为0.037μg/g和0.043μg/g。The citrinin binding capacity of muscle protein from bullfrog(Rana catesbeiana)immunized with citrinin-bovine serum albumin(BAS)conjugate emulsified with Freund’s complete adjuvant was investigated.The protein with high citrinin binding capacity was embedded into sodium alginate microspheres.The optimum conditions were determined as follows:immunogen dosage 6.4 mg,sodium alginate concentration 4 g/100 mL,CaCl2 concentration 5 g/100 mL,glutaraldehyde concentration 0.2%,and crosslinking time 2 h.Two milliliter of the crude citrinin extract from red rice was passed through a column packed with 2 g of sodium alginate microspheres followed by washing with 20 mL of ultra-pure water to remove the impurities,and good chromatogram was obtained without interfering peaks compared with commercial immunoaffinity column.The recovery of citrinin standard solution eluted with 5 mL of 70%acetonitrile and trifluoroacetic acid mixture was 86.7%.The column capacity was 0.101μg/g.The content of citrinin in red rice was determined to be 0.037 and 0.043μg/g using the column developed in this study and commercial immunoaffinity column,respectively.However,further study should be done to improve the preparation process for better product performance.

关 键 词：橘霉素 牛蛙 免疫亲和柱 

分 类 号：TS201.3[轻工技术与工程—食品科学]