摘要
用酸性染料为探针对蛋白质的定量已应用于临床医学及生命科学的研究中, 本文利用分光光度法研究溴酚蓝(BPB)与牛血清白蛋白(BSA)、人血清白蛋白(HSA)的反应. 在pH=2.79的Britton-Robinson缓冲溶液中, 血清白蛋白与溴酚蓝通过分子间力形成蓝色复合物使吸光度增加. λ=605 nm时, εBSA=5.20×105 L·(mol·cm)-1, εHSA=5.12×105 L·(mol·cm)-1;BSA在0～78.0 mg·L-1, HSA在0～81.6 mg·L-1范围内服从比尔定律. 检出限分别为BSA: 2.93 mg*L-1, HSA:3.12 mg·L-1. 对7个人血清蛋白总量平行6次测定, 相对标准偏差1.2%～3.1%, 回收率91.9%～105.6%.
A acidic dyes have been applied to the assay of proteins in the fields of medicine and life science. The reactions of human serum albumen (HSA) and bovine serum albumen (BSA) with bromophenol blue have been studied. Both BSA and HSA can form blue complexes, which increase the absorbance. When wavelength was set at 605 nm, epsilon(BSA) = 5.20 X 10(5) L-.(mol(.)cm)(-1), epsilon(HSA) = 5.12 x 10(5) L-.(mol(.)cm)(-1). In the range of 0-78.0 mg(.)L(-1) for BSA and 0-81.6 mg(.)L(-1) for BSA, there exists a linear relationship between protein concentration and absorbance. The detection limits were 2.93 mg(.)L(-1) for BSA and 3.12 mg(.)L(-1) for HSA. The relative standard deviation and the recovery of the method for the determination of total proteins for seven human serum samples were 1.2 % -3.1 % and 91.9%-105.6%, respectively. The results obtained by this method were in agreement with those by Biuret method. The method proposed in this paper is sensitive, accurate and tolerant to many foreign substances. Moreover, all the reagents were stable and the performance was easy to carry out.
出处
《光谱学与光谱分析》
SCIE
EI
CAS
CSCD
北大核心
2003年第5期999-1001,共3页
Spectroscopy and Spectral Analysis
基金
河北省教育厅博士基金资助
