An amperometric biosensor based on lactate oxidase immobilized in laponite-chitosan hydrogel on a glassy carbon electrode. Application to the analysis of L-lactate in food samples

摘要

A biosensor based on the immobilization of lactate oxidase (LOx) on a glassy carbon electrode modified with laponite/chitosan hydrogels for the quantification of L-lactate in alcoholic beverages and dairy products is presented. Ferrocene-methanol (FcMe) is used as artificial mediator. The purpose of this work is to determine the best hydrogel composition from the analytical point of view. The characterization of the hydrogels was carried out by CV, amperometry and EIS. According to permeabilities and charge transfer resistances for ferrocyanide (used as molecular probe) as well as the enzymatic behavior of the enzyme for L-lactate, the best laponite/chitosan mass ratio found was 25/50. The distinct features of the bioelec-trode are its long stability, its ability to reject or minimize most interferents including ascorbic acid, and its excellent analytical response, which allowed the reduction of the enzyme content below 0.5 U, for a sensitivity of (0.326±0.003)Acm~(-2) M~(-1), with a time response lower than 5s and a detection limit of (3.8 ±0.2) x 10~(-6) M. Our L-lactate biosensor was validated by comparison with a standard spectroscopic method.