A reagentless acetaldehyde sensor is designed based on the co-entrapment of aldehyde dehydrogenase (AIDH), NADH oxidase and high molecular weight NADs (NAD-dextran and NAD-PEG) in a polyvinyl alcohol) bearing stryrylpyridinium groups (PVA-SbQ) matrix. The amperometric detection is based on hydrogen peroxide oxidation by applying a potential difference of 550 mV between two platinum electrodes, equivalent toca.600 mVvs.SCE. The sensitivity is close to 2 mA 1 mol−1, the linear range extends from 0.5 μM to 330 μM acetaldehyde and the response time is lower than 3 minutes. More than 80 reproducible measurements can be performed without cofactor addition, indicating a good operational stability. Acetaldehyde concentration in alcoholic beverages is determined and validation of the results is made using a classical spectrophotometric method.