Abstract
An integrated amperometric d-lactic acid biosensor involving a gold film deposited by sputtering on a stainless steel disk electrode where the enzymes d-lactic acid dehydrogenase (DLDH) and diaphorase (DP) as well as the redox mediator tetrathiafulvalene (TTF) are coimmobilized by using a dialysis membrane, is reported in this work. Amperometry in stirred solutions at a detection potential of +0.15V (vs Ag/AgCl reference electrode) provided a linear calibration plot for d-lactic acid over the 1.0×10−4 to 3.8×10−3g L−1 concentration range, with a limit of detection of 3.1×10−5g L−1. The usefulness of the biosensor was demonstrated by determining d-lactic acid in beer samples with good results. Additionally, the biosensor was implemented together with a commercial l−lactic amperometric biosensor in a semiautomatic flow-injection analysis (FIA) system able to perform a rapid and simple stereo-specific determination of d- and d-lactic without a previous separation step. The operational characteristics of the biosensors under flow conditions were evaluated and its applicability was demonstrated through the simultaneous determination of both enantiomers in beer samples.
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