Chemiluminescence Flow Biosensor System for Cholesterol with Immobilized Reagents

Abstract

A chemiluminescence biosensor combined with flow injection analysis has been developed for determining cholesterol. Cholesterol oxidase was immobilized onto amine-modified silica gel via glutaraldehyde activation and packed in a column. The analytical reagents, including luminol and ferricyanide, were electrostatically coimmobilized on an anion-exchange column. Cholesterol was sensed by the CL reaction between hydrogen peroxide released from the enzymatic reaction and luminol and ferricyanide, which were released from a column with immobilized reagents by elution. The calibration graph was linear over the range 5×10^-6 to 1×10^-4 g ml^-1 and the detection limit was 5×10^-6 g ml^-1. The pro-posed method has been successfully applied to the determination of cholesterol in human serum.