Hemin catalytic fluorescence reaction with pyronine B as substrate for the determination of glucose

作者：Chen, LH; Huang, F; Liu, LZ; Shen, HX^*

来源：Chinese Journal of Analytical Chemistry, 2003, 31(10): 1237-1239.

• pyronine B
• hemin
• mimic enzyme
• glucose
• fluorogenic substrate

Pyronine B (tetraethyldiaminoxanthenyl chloride, PB) was used as a fluorogenic substrate for the determination of hydrogen peroxide and glucose based on the catalytic effect of hemin as mimic-enzyme of horseradish peroxidase. The fluorescence of pyronine B was quenched by hydrogen peroxide in the presence of hemin at pH 5.2. The steady-state catalytic rate depends upon mimic-enzyme and substrate concentrations, and the Michaelis-menten parameters K-m, V-max and K-cat are 1.4 x 10(-5) mol/L, 1.9 x 10(-6) mol/L.s and 25.6/s, respectively. Under optimum conditions, linear relationship between the fluorescence quenching (F-0/F) and the concentration of hydrogen peroxide was observed in the range of 0.0 to 7.2 x 10(-7) mol/L. The limit of detection (3sigma) was 8.0 x 10(-9) mol/L. By coupling with glucose oxidase-catalytic reaction, glucose was quantified in the linear range of 0.0 to 1.0 x 10(-5) mol/L with the limit of detection (3sigma) of 3.3 x 10(-8) mol/L. The determination results of glucoses in human serum are in agreement with those by the phenol-4-aminoantipyrine method.

• 出版日期2003-10
• 单位吉首大学; 南开大学