title: Quantifying ethanol by high performance liquid chromatography with precolumn enzymatic conversion and derivatization with fluorimetric detection.
abstract: We quantified ethanol by measurement of the subsequent increase in acetaldehyde after reaction with alcohol dehydrogenase and nicotinamide adenine dinucleotide (ADH-NAD) with a fluorimetric HPLC method. Ethanol standards ranging from 0.3 to 200 mg/dl were investigated and the limit of quantitation of the fluorimetric HPLC method was found to be 6 mg/dl. The accuracy of the HPLC method was assessed by assaying blood samples containing 6-200 mg/dl of ethanol and comparing its results to those of the ADH-NAD enzymatic method (r2 = 0.993). The coefficients of variation for intraassay (assayed ten times) and interassay (assayed on 7 consecutive days) were 6.7% and 9.3% for blood samples containing 50 mg/dl of ethanol and 4.0% and 17.9% for blood samples containing 200 mg/dl of ethanol. The blood ethanol concentrations of a volunteer after a pulse of 0.3 g/kg of ethanol determined with the described HPLC method were correlated to the results from the ADH-NAD enzymatic method (r2 = 0.986). In conclusion, the fluorimetric HPLC method for measurement of ethanol here described is of potential clinical utility.
authors.affiliation: Sansum Medical Research Foundation, Santa Barbara, CA 93105.
authors.forename: H M
authors.initials: HM
authors.lastname: Chen
authors.forename: C M
authors.initials: CM
authors.lastname: Peterson
mesh_terms: D000426:Alcohol Dehydrogenase / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D000431:Ethanol / Q000097:blood*; D006801:Humans; D009243:NAD / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D009243:NAD; D000431:Ethanol; D000426:Alcohol Dehydrogenase