摘要
采用电化学方法和紫外-可见光谱法对邻氨基酚(OAP)-H2O2-辣根过氧化物酶(HRP)酶联免疫分析体系的反应产物进行了较为详细地研究。紫外-可见光谱实验表明HRP的加入极大地催化了H2O2氧化OAP的反应。循环伏安实验结果表明,其酶促产物在玻碳电极上发生可逆的氧化还原反应,峰电流与扫描速率的平方根成线性关系;微分脉冲伏安曲线表明酶促产物在-0.336V左右(pH8.0B—R缓冲溶液中)产生了1个峰形良好的还原峰,峰电流随HRP浓度的增大而增大,借助此还原电流可用于测定HRP。用微分脉冲伏安法对酶促产物的测定条件进行了优化。在最佳条件下测定游离HRP的线性范围为8.0×10^-11~4.0×10^-9g/mL。检出限为6.9×10^-11g/mL。
A voltammetric detection of horseradish peroxidase (HRP) using glassy carbon electrode (GCE) was investigated based on the o-aminophenol(OAP) -H2O2 -HRP coupling reaction system. The product of OAP oxidized by H2O2 in the presence of HRP can be reduced at - 0. 336 V ( vs. Ag/AgCl) on GCE and the differential pulse vohammetry (DPV) was chosen to detect the enzymaticgenerated product. The conditions for HRP electrochemical detection were carefully studied with a CHI832 electrochemical analyzer. The reduction peak current was linear with the HRP concentration ranging from 8.0 × 10 ^-11 to 4. 0 × 10^ -9 g/mL and the detection limit for HRP was 6.9 × 10^-11 g/mL.
出处
《分析测试学报》
CAS
CSCD
北大核心
2006年第2期10-12,共3页
Journal of Instrumental Analysis
基金
国家自然科学基金资助项目(20375020)
关键词
辣根过氧化物酶
邻氨基酚
电化学
酶促反应
Horseradish peroxidase
o-Aminophenol
Electrochemistry
Enzymatic reaction