苦胺酸偶氮变色酸与血清蛋白质相互作用的分光光度研究

Spectrophotometric Study on the Interaction Between Picramazochrom and Serum Proteins

在 p H1 .2的 Clark- Lubs缓冲介质中 ,苦胺酸偶氮变色酸与血清蛋白质作用在室温下能迅速结合形成紫色的复合物 ,其最大吸收波长为 62 5 nm,比苦胺酸偶氮变色酸本身红移了 1 0 0 nm。用分光光度法研究了该结合反应的最佳条件 ,建立了一个测定蛋白质的新方法 ,测定的表观摩尔吸光系数 ε62 5达 2 .72 8× 1 0 5L/mol·cm(牛血清白蛋白 ) ,线性范围为 1 0～ 1 5 0 mg/L。所拟方法简便、快速、选择性好、灵敏度高 ,应用于人血清中总蛋白的测定 ,结果与考马斯亮蓝 G- 2 5 0法一致。

It was found that Picramazochrom can react with proteins forming a violet complex in the Clark Lubs medium at pH1.2 and room temperature, which gives the absorption peak at 625 nm with 100 nm of bathochromic shift in from that of Picramazochrom. A molar absorptivity ε 625 2.728×10 5 L/(mol·cm) and linearity range of 10~150 mg/L for BSA were determined. The optimum reaction conditions were investigated, and a new method for the determination of proteins was developed based on this binding reaction. The proposed method, which is easy, rapid, stable, and of good selectivity and high sensitivity, has been used for the determination of total proteins in human serum samples . The results obtained by this method were agreed with those obtained by the Coomassie brillant blue G 250 method.