Unveiling hidden threats: Novel detection of acetaldehyde dehydrogenase inhibitors in everyday foods

Ethanol metabolism relies on acetaldehyde dehydrogenase (ALDH) to convert toxic acetaldehyde into harmless acetic acid. Some food-derived ALDH inhibitors can disrupt this process, raising health risks from alcohol consumption. Thus, identifying these inhibitors quickly and accurately in foods is crucial. Here we developed a novel analytical technique combining biosensing with affinity chromatography to detect ALDH inhibitors. The new concept of ALDH-Acetaldehyde-β-Nicotinamide adenine dinucleotide hydrate (NAD + ) based colorimetric sensing protocol exhibited good linearity within an ALDH activity range of 1.465–4.395 U/mL. The optimized detection procedure uses 0.1 % acetaldehyde, 10 mM NAD + , a 3:3 WST-8 to methoxy-PMS ratio, and two 30-min reactions at 37 °C. The efficacy of the method was validated with both negative and positive drugs and then applied to screen ALDH inhibitors in eleven foods, identifying soybean [ Glycine max (L.) merr.] as a significant inhibitory. Within soybean, daidzin was found to have an IC 50 of 194.09 μM. Molecular docking studies reveal that daidzin effectively binds to ALDH's active site. The detection method developed here is simple, highly sensitive, and reproducible, providing an efficient approach for identifying and screening new enzyme inhibitors in foods. • A novel colorimetric biosensor for rapid screening of ALDH inhibitor was established. • Established affinity biosensing can easily and precisely screen ALDH inhibitors. • Specific ALDH inhibitors contained in food were identified using LC-MS. • Daidzin exhibited significant ALDH inhibition effects with an IC 50 of 194.09 μM. • Avoid the intake of soybeans and related products when drinking alcohol.