Spectrophotometric Determination of Proteins by Using Nitrosulfophenol C

蛋白质的定量分析是生化研究、临床化验和食品检验等领域经常涉及的内容.以有机小分子作光谱探针测定蛋白质,如甲基橙[1,2]、考马斯亮蓝G-250[3]、溴酚蓝[4]、溴甲酚绿[5]和偶氮胂Ⅲ[6]等已得到研究.本文研究表明,在室温条件下,硝基磺酚C能与蛋白质在pH3.4的B-R缓冲介质中生成复合物,最大吸收波长为675nm,比试剂本身红移100nm.在确定的最佳实验条件下,制定了8种蛋白质的工作曲线,线性范围为0～140mg/L.测定了人血清样品中蛋白质的含量,与经典的考马斯亮蓝G-250方法结果一致,回收率令人满意.本方法灵敏度高,重现性好,操作简便,基本无干扰.In this paper, it is found firstly that the nitrosulfophenol C can react with proteins to form a pure blue complex in Britton-Robinson buffer solution at pH 3.4 and room temperature, which gives maximum absorption peak at 675 nm with 100 nm to red shift compared to that of nitrosulfophenol C, According to studies on optimum conditions, a new spectrophotometric method for determination of proteins was established, The standard working curves of eight kinds of proteins with larger linear ranges and higher sensitivity were determined. The new method was successfully applied to the determination of protein in human serum samples with satisfactory results in agreement with those of the classical coomassie brilliant blue G-250 (CBB G-250) method.中国科学院资助项目SCI(E)EI中文核心期刊要目总览(PKU)中国科学引文数据库(CSCD)322212