[一种辣根过氧化物酶的新型催化分光光度测定法]

[A novel catalytic spectrophotometric determination for horseradish peroxidase].

作者信息

Wei Y F, Yan H T

机构信息

Department of Chemistry, Northwest University, Xi'an 710069, China.

出版信息

Guang Pu Xue Yu Guang Pu Fen Xi. 2001 Oct;21(5):704-6.

PMID:12945338

Abstract

A new catalytic-kinetics spectrophotometric method for the determination of horseradish peroxidase was developed. It was based on the catalytic effect of horseradish peroxidase (HRP) on the coloration reaction, in which the reduced rhodamine B was oxidized by H2O2 in pH 6.80 phosphate buffer. It was found that reduced rhodamine B stocking solution could be steady in 0.25% beta-CD solution. The kinetic behavior of the reaction and the effects of some experimental conditions were investigated and discussed in detail. The method has been applied to determine HRP with a satisfactory result. The calibration curve is linear over the range 15-250 pg.10 mL-1 of HRP (r = 0.9989). The limit of detection was 12 pg.10 mL-1 and the relative standard derivative RSD is 4.25% by determining for 20 pg.10 mL-1 HRP (n = 10).

摘要

建立了一种测定辣根过氧化物酶的催化动力学分光光度新方法。该方法基于辣根过氧化物酶（HRP）对显色反应的催化作用，即在pH 6.80的磷酸盐缓冲溶液中，过氧化氢将还原罗丹明B氧化。研究发现，还原罗丹明B储备液在0.25%的β-环糊精溶液中可保持稳定。详细研究并讨论了反应的动力学行为及一些实验条件的影响。该方法已用于测定HRP，结果令人满意。HRP在15 - 250 pg·10 mL-1范围内校准曲线呈线性（r = 0.9989）。通过测定20 pg·10 mL-1的HRP（n = 10），检测限为12 pg·10 mL-1，相对标准偏差RSD为4.25%。