催化分光光度法测定血红蛋白  

Catalytic Spectrophotometric Determination of Hemoglobin

作　　者：陈亚红[1] 张建夫[1] 翟向娜[1] 

机构地区：[1]周口师范学院化学系,周口466000

出　　处：《理化检验（化学分册）》2008年第10期957-959,共3页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

基　　金：河南省教育厅自然科学基金资助项目(2007150048)

摘　　要：基于血红蛋白的过氧化物酶特性，建立了一种测定血红蛋白的催化动力学光度分析法。在pH9．5的缓冲溶液中，血红蛋白对过氧化氢氧化酸性铬蓝K反应体系的褪色作用明显加速。在最佳试验条件下，由于褪色作用的加速所引起的吸光度下降（△A）与血红蛋白的浓度在7．0×10^-10～9．0×10^-8mol·L^-1范围内呈线性关系。表观摩尔吸光率为3．9×10^-8L·mol^-1·cm^-1，检出限为1．7×10^-1mol·L^-1。用于尿液中血红蛋白含量的测定，测定结果的相对标准偏差（，2—5）在1．9%～4．2％之间，回收率在96．0％～102．0％之间。对此酶催化反应的机理也作了探讨。Based on its peroxidase property, a new method of catalytic spectrophotometric determination of hemoglobin （Hb） was proposed. In an ammoniacal buffer medium of pH 9. 5, oxidation of acid chrome blue K （ACBK） by hydrogen peroxide was catalyzed by hemoglobin, leading to acceleration of color-fading of the reaction system. Under the optimized reaction condition, the magnitude of decrease in absorbance （AA） due to catalytic effect was found to keep a linear relationship with the concentration of Hb in the range of 7. 0 × 10^-10- 9. 0 × 10^-8 mol · L^-1, with its detection limit （3Sb/k） of 1.7× 10^-11mol · L^-1, and apparent molar absorptivity of 3.9×10^8L · mol^-1 ·cm^-1. In application of the proposed method to the determination of Hb in urine, values of RSD＇s （n=5） obtained were in the range of 1.9%-4. 2%, and values of recovery were in the range of 96. 0%-102. 0%. A brief discussion on the mechanism of the enzyme catalysis was also given.

关 键 词：催化分光光度法 血红蛋白 酸性铬蓝K 过氧化氢 

分 类 号：O657.31[理学—分析化学]