Enzyme-less amperometric biosensor for L-ascorbate using poly-L- histidine-copper complex as an alternative biocatalyst

Abstract

A poly-L-histidine(PLH)-copper(II) complex can be used as an alternative biocatalyst in an O2 detection-type amperometric enzyme-less L-ascorbate (AsA) sensor. The PLH-Cu(II)membrane was simply prepared by entrapping the PLH in polyacrylamide gel and subsequent treatment of the gel with CuCl2 solution. This enzyme-less biosensor can be used over a relatively wide pH region from 4 to 11 and enables precise determination of AsA (RSD less than 3%, n = 10) at pH 7.0. The fundamental performance characteristics (sensitivity, response time, and linear range) of this PLH-Cu(II)-based sensor is comparable to those of a native ascorbate oxidase-based sensor. Unfortunately, the selectivity is inherently rather low and, as a result, the response was degraded in the presence of higher concentrations (more than mM order) of quinones. However, reducing sugars caused no interference and the sensor could be used to detect AsA in some fruits and drinks. This enzyme- less sensor has excellent stability for at least 3 months of repeated analysis (more than 300 samples) without loss of ordinal activity.