Quantifying ethanol by high performance liquid chromatography with precolumn enzymatic conversion and derivatization with fluorimetric detection.
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.
Chen
H M
HM
Sansum Medical Research Foundation, Santa Barbara, CA 93105.
Peterson
C M
CM
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
D009243:NAD; D000431:Ethanol; D000426:Alcohol Dehydrogenase