Assay of cholinesterase using a proenhancer of the luminol–H₂O₂–horseradish peroxidase reaction

Abstract

2-Naphthyl acetate acts as a pro-enhancer of the luminol-H2O2-horseradish peroxidase reaction. Cholinesterase hydrolyses the bound acetyl group and produces 2-naphthol, and this compound is an enhancer of the chemiluminescent reaction. We studied the kinetics of chemiluminescent emission and the influence of 2-naphthyl acetate and cholinesterase enzyme concentrations. The cholinesterase concentration versus chemiluminescence intensity maximum was linear for cholinesterase between 0 and 181 microU/mL, with a detection limit of 8 microU/mL and a relative standard deviation of 9.5% (n = 3), for a sample containing 90.67 microU/mL of cholinesterase.

A Novel Method for Time‐Resolved Fluorimetric Determination and Imaging of the Activity of Peroxidase, and Its Application to an Enzyme‐Linked Immunosorbent Assay

“…Some of these products are suitable for chromogenic, fluorogenic or other signal-generating purposes. As a result, kinetic assays have been developed based on spectrophotometry/reflectometry, [28,29] fluorimetry, [30,31] chemiluminescence, [32][33][34] electrochemiluminescence, [35] and electroanalysis. [36,37] Among the fluorimetric methods, those based on time-resolution [38] (often referred to as time-resolved or gated assays) are the most sensitive tools in biological assays.…”