ZnO nanowire-based fluorometric enzymatic assays for lactate and cholesterol

Abstract

A rapid fluorometric method is described for the determination of lactate and cholesterol by using ZnO nanowires (ZnO NWs). The assay is based on the detection of the hydrogen peroxide generated during the enzymatic reactions of the oxidation of lactate or cholesterol. Taking advantage of the electrostatic interactions between the enzymes and the ZnO NWs, two bioconjugates were prepared by mixing the nanomaterial and the enzymes, viz. lactate oxidase (LOx) or cholesterol oxidase (ChOx). The enzymatically generated hydrogen peroxide quenches the fluorescence of the ZnO NWs, which have emission peaks at 384 nm and at 520 nm under 330 nm photoexcitation. H₂O₂ quenches the 520 nm band more strongly. Response is linear up to 1.9 μM lactate concentration, and up to 1.1 μM cholesterol concentration. Relative standard deviation was found to be 5%. The detection limits for lactate and cholesterol are 0.54 and 0.24 μM, respectively. Graphical abstractSchematic representation of fluorescence assay based on ZnO nanowires photoluminiscence for lactate and colesterol detection.

Keywords: Cholesterol oxidase; Collisional quenching mechanism; Enzymatic reaction; Fluorescence; Human serum sample; Hydrogen peroxide; Indirect optical method; Lactate oxidase; Photoluminescence; ZnO NWs-enzyme bioconjugate.