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旨在建立一种蛋白质印迹模板提取UV法测定大豆中蛋白质的新方法。以牛血清蛋白(BSA)为目标分子,甲基丙烯酸(MAA)为功能单体,乙二醇二甲基丙烯酸酯(EDGMA)为交联剂,偶氮二异丁腈(AIBN)为引发剂制备了蛋白质分子印迹模板,研究了模板对BSA的吸附性能,并作为提取材料,提取了大豆中的蛋白质,洗脱后用紫外法测定大豆中的蛋白质含量。结果表明:蛋白质模板吸附和洗脱的最佳pH值分别为5.3和5.6,最佳吸附时间为2 h,最大吸附量为72 mg/g。3种大豆样品中蛋白质的含量分别为364、427、405 mg/g,回收率为93.5%~108.0%。结果表明,制备的蛋白质印迹模板可作为提取材料,测定大豆中的蛋白质含量。
Aim To establish a new Western blot template extraction UV method for determination of protein in soybeans. Bovine serum albumin (BSA) as the target molecule, methacrylic acid (MAA) as the functional monomer, ethylene glycol dimethacrylate (EDGMA) as the crosslinking agent and azobisisobutyronitrile (AIBN) as the initiator The protein molecularly imprinted template was prepared, the adsorption of BSA to the template was studied, and the protein in soybean was extracted as the extraction material. The content of protein in soybean was determined by UV. The results showed that the optimal pH values for the adsorption and elution of protein templates were 5.3 and 5.6, respectively. The optimum adsorption time was 2 h and the maximum adsorption capacity was 72 mg / g. The contents of protein in the three soybean samples were 364, 427 and 405 mg / g, respectively. The recoveries ranged from 93.5% to 108.0%. The results show that the prepared Western blot template can be used as extraction material to determine the protein content in soybean.