QUANTIFYING ETHANOL BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH PRECOLUMN ENZYMATIC CONVERSION AND DERIVATIZATION WITH FLUOROMETRIC DETECTION

We quantified ethanol by measurement of the subsequent increase in ace taldehyde after reaction with alcohol dehydrogenase and nicotinamide a denine dinucleotide (ADH-NAD) with a fluorimetric HPLC method. Ethanol standards ranging from 0.3 to 200 mg/dl were investigated and the lim it of quantitation of the fluorimetric HPLC method was found to be 6 m g/dl. The accuracy of the HPLC method was assessed by assaying blood s amples containing 6-200 mg/dl of ethanol and comparing its results to those of the ADH-NAD enzymatic method (r(2) = 0.993). The coefficients of variation for intraassay (assayed ten times) and interassay (assay ed on 7 consecutive days) were 6.7% and 9.3% for blood samples contain ing 50 mg/dl of ethanol and 4.0% and 17.9% for blood samples containin g 200 mg/dl of ethanol. The blood ethanol concentrations of a voluntee r after a pulse of 0.3 g/kg of ethanol determined with the described H PLC method were correlated to the results from the ADH-NAD enzymatic m ethod (r(2) = 0.986). In conclusion, the fluorimetric HPLC method for measurement of ethanol here described is of potential clinical utility .