We present a case of a death of a diabetic man where the concentration of ethanol in post-mortem blood rose from 0.4 g/l 2 days after autopsy to 3.5 g/l 10 days after autopsy. The presence of fluoride ions in this blood sample was determined with ion chromatography and verified that fluoride ions were added to the vials. The concentrations of free fluoride, corresponding to 0.21 and 0.25% w/v potassium fluoride in blood and urine, respectively, were somewhat lower than the recommended 1% w/v. However, the amount of fluoride ions bound to calcium, proteins and other compounds in the samples is unknown. The blood sample was also subject to microbiological examination, which revealed growth of bacteria. In addition, a very high concentration of glucose was found in vitreous humour from the deceased. To determine whether the ethanol detected at the first analysis was of ante-mortem origin, ethyl glucuronide was analysed. Its absence, in the blood as well as the urine sample, strongly supported the theory that, in this case, all the ethanol detected was formed post-mortem. This case showed that ethanol may be formed in vitro at a very high concentration, despite the verified presence of fluoride ions. Possible reasons for this unusual formation of ethanol were the abundant presence of bacteria, a high level of glucose and, possibly, an insufficient amount of fluoride added to the vials.

No abstract

A sensitive, fast, simple, and high-throughput enzymatic method for the quantification of ethanol in whole blood (blood) on Hitachi 917 is presented. Alcohol dehydrogenase (ADH) oxidizes ethanol to acetaldehyde using the coenzyme nicotinamide adenine dinucleotide (NAD), which is concurrently reduced to form NADH. Method development was performed with the aid of factorial design, varying pH, and concentrations of NAD+ and ADH. The linear range increased and reaction end point decreased with increasing NAD+ concentration and pH. The method was linear in the concentration range 0.0024-0.4220 g/dL. The limits of detection and quantification were 0.0007 g/dL and 0.0024 g/dL, respectively. Relative standard deviations for the repeatability and within-laboratory reproducibility were in the ranges 0.7-5.7% and 1.6-8.9%, respectively. The correlation coefficient when compared with headspace gas chromatography-flame ionization detection methods was 0.9903. Analysis of authentic positive blood specimens gave results that were slightly lower than those of the reference method.