Botrytis cinerea is the causative agent of gray mold disease, which causes considerable economic losses to winemakers. The extent of gray mold infection of winegrapes is commonly visually estimated, a method that is prone to assessor bias. Here, we used rapid and simple enzyme-based screening consisting of carbon-electrode, screen-printed amperometric biosensors to estimate gluconic acid and glycerol concentration in winegrapes infected with different degrees of B. cinerea. The lower limits of quantification of the screen-printed amperometric biosensors were 3 mg/L for gluconic acid (corresponding to an infection rate of less than 1%) and 35 mg/L for glycerol; the response times with a flow rate of 0.5 mL/min were in a range of 0.5 to 2 min in the linear ranges of the two assays. This study demonstrates the efficacy of amperometric biosensors for rapid analysis of gluconic acid and glycerol in grapes. The …