Fluorescein Chemiluminescent Assay of Glucose in Serum Using Glucose Oxidase and Horseradish Peroxidase

Abstract

Glucose in serum was successfully determined by the simultaneous coupling of a glucose-glucose oxidase (GOD) reaction with a fluorescein (FL)-chemiluminescent (CL) reaction catalyzed by horseradish peroxidase. The FL-CL reaction could be conducted at pH 7.0, thus allowing the in situ detection of hydrogen peroxide generated during the glucose-GOD reaction. The CL emission lasted for about 3min, and the calibration curve was linear over a glucose concentration range of 5.0×10^-8to 1.0×10^-5M. The relative standard deviation at 5.0×10^-7M glucose was 1.8% (n=5). The present method was not subject to any interference from components normally present in serum. The results of glucose assays in two control sera were in good agreement with those conducted by the colorimetric method.