基于血红蛋白的过氧化物酶特性测定其在血浆中游离态含量  

Determination of Free Hemoglobin in Plasma Based on Its Peroxidatic Characteristic

作　　者：王全林[1] 吕功煊[1] 

机构地区：[1]中国科学院兰州化学物理研究所羰基合成与选择氧化国家重点实验室,兰州730000

出　　处：《分析化学》2003年第8期945-949,共5页Chinese Journal of Analytical Chemistry

基　　金：国家基础研究发展规划资助项目 (G2 0 0 0 0 2 6 4)

摘　　要：基于血红蛋白 (Hb)的过氧化物酶特性 ,研究了以铬黑T(EBT)作为其氢供体底物时的酶动力学行为。实验测定了Hb作为辣根过氧化物酶 (HRP)的替代物的类酶催化米氏常数和米氏速率。对Hb催化H2 O2 氧化铬黑T的机理作了探讨。建立了测定Hb的催化动力学光度法 ,方法的线性范围为 2× 10 - 8mol/L～ 2× 10 - 6mol/L ,检出限为 3.6× 10 - 9mol/L。用于血浆中游离Hb的测定 ,结果满意。Based on peroxdatic characteristic of hemoglobin, enzyme kinetic behavior was studied with eriochrome black T as hydrogen donor substrate. Reaction velocity depends upon hemoglobin and substrate concentrations, and a Michaelis-Menten K-m value and a V-m value were measured at pH 7.5. The mechanism that eriochrome black T was cleaved into aromicdiazonium ions by catalytic effect of hemoglobin was discussed. A catalytic-kinetics spectrophotometric method was proposed for the determination of hemoglobin. The initial rate of fad reaction of eriochrome black T at the wavelength of 615 nm was monitored, permitting a detection limit of 3.6 x 10(-9) mol/L hemoglobin. A linear calibration graph was obtained over the hemoglobin concentration range of 2 x 10(-8) similar to 2 x 10(-6) mol/L, and the relative standard deviation at a hemoglobin concentration of 7 x 10(-7) mol/L was 2.1% (n = 7). Satisfied results were obtained in the determination of hemoglobin in plasma samples by this method.

关 键 词：血红蛋白 过氧化物酶 测定 血浆 游离态 含量 分光光度法 催化动力学 电分析化学 

分 类 号：O657.3[理学—分析化学]