Liquid chromatographic determination of ethyl alcohol in body fluids

A high-performance liquid chromatographic technique for ethyl alcohol deter mination in body fluids is proposed. Ethyl alcohol is quantitatively conver ted into acetaldehyde-phenylhydrazone by oxidation in the presence of alcoh ol dehydrogenase, nicotinamide-adenine dinucleotide and phenylhydrazine. Th e derivative is suitable for reversed-phase liquid chromatography and ultra violet detection at 276 nm. The limits of linearity, detection and quantifi cation as well as accuracy and reproducibility were investigated in water, serum and whole blood. Analytical responses were linear within the 0.008 to 5 g/l range, and the limit of quantification was 0.02 g/l both in aqueous standard and in biological matrix assays. Mean analytical recovery of ethyl alcohol in blood serum averaged 98.2+/-4.2%, imprecision (CV%) at 0.80 g/l was 2.2%, and the limit of quantification was 0.02 g/l. Serum concentratio ns of persons that avoided alcoholic beverages for a week were less than th e limit of quantification. Ethyl alcohol concentrations in serum and whole blood compared well with those obtained by headspace gas chromatography. Th is simple and reliable procedure, which was also used for a urine assay, co uld be suitable for validation of the screening procedures used to monitor ethanol abuse. (C) 1999 Elsevier Science B.V. All rights reserved.