An approach to the glucose determination by amperometric biosensing in wine industry applications is presented. Integrated screen-printed biosensors based on horseradish peroxidase (HRP) and glucose oxidase (GOx) have been developed. The experimental design methodology has been used to find the optimum conditions of the experimental variables, in such a way that a chronoamperometric response specific for glucose was recorded. Under these conditions, repeatability and reproducibility of the modified electrodes have been analyzed. The detection limit for glucose has been calculated taking into account the probability of false positive (alpha) and negative (beta), reaching a medium value of 4.37+/-0.21 micromol dm-3 (alpha=beta=0.05, and a replicate n=4). The biosensor was applied to the determination of glucose in white wine samples.