Validation of an enzymatic method for the determination of trace levels of ethanol

The validation of an enzymatic method for the determination of trace levels of ethanol is described. The method, which involves the use of alcohol dehydrogenase (ADH) and nicotinamide adenine dinucleotide (NAD(+)), was validated according to USP 23 requirements. The validation parameters, including linearity and range, accuracy (recovery), precision, ruggedness, limit of detection (LOD), and limit of quantitation (LOQ), were established satisfactorily. The method is specific and selective for ethanol except in the presence of propyl, isopropyl, or butyl alcohol.