Published: Journal of Analytical Toxicology, Volume 20, Number 2, March/April 1996, pp. 141-142.

Disclosing Recent Drinking after Alcohol Has Been Cleared from the Body
A.W. Jones and A. Helander

To the Editor:
Excessive drinking is a major cause of accidents within the home, at work, and on the roads. In addition to the acute effects of alcohol on a person’s performance and behavior, the aftereffects of heavy drinking (hangover) are often associated with impairment of sensory and motor functions (1,2). During a hangover, people are likely to perform worse in skilled tasks for many hours, even after alcohol has disappeared from the body (3). Accidents in industry and elsewhere might be caused by diminished attention, fatigue, or some other physiological disturbance associated with the hangover state (4). A way to detect recent drinking after alcohol has been cleared from the body would have considerable importance when accidents are investigated by police and insurance organizations. We report here a biochemical test to disclose if a person has recently been engaged in drinking after the alcohol has been metabolized.

Ethanol is oxidized in the liver to acetaldehyde by the action of alcohol dehydrogenase (ADH), and this primary metabolite is rapidly converted into acetate by aldehyde dehydrogenase (ALDH). During metabolism of ethanol, the ratio of NADH/NAD+ in the hepatocyte increases appreciably, and many NAD-dependent biochemical reactions are disturbed. Moreover, ALDH is also involved in the biotransformation of biogenic aldehydes, for example, the intermediate formed when serotonin (5-hydroxytryptamine) is deaminated by monoamine oxidase. This biogenic aldehyde is normally oxidized by ALDH to produce the end-product, 5-hydroxyindoleacetic acid (5HIAA), but a small fraction is also reduced to the alcohol metabolite, 5-hydroxytryptophol (5HTOL). Under normal circumstances and without the intake of alcohol, the urinary ratio of 5HTOL/5HIAA is extremely small (5). When the body is engaged in the metabolism of ethanol, the ratio of 5HTOL/5HIAA increases appreciably in blood and urine (5). This follows because the raised NADH/NAD+ ratio favors reduction of aldehydes to produce alcohol metabolites, and furthermore, ALDH is engaged in the disposal of acetaldehyde derived from ethanol. More importantly, after drinking alcohol, the 5HTOL/5HIAA ratio increases in a dose-dependent manner and remains elevated for 5–15 h after blood and urine alcohol concentrations reach endogenous levels (6).

A male subject (49 years old; 73 kg) was engaged in heavy drinking on three separate occasions, consuming 100–150 g ethanol during the evening and the next morning suffering from hangover. After awakening, his blood alcohol concentration (BAC) was tested indirectly by analysis of breath with Alcolmeter S-D2 (Lion Laboratories PLC., Barry, Wales), and samples of the first, second, and on one occasion the third urinary void were collected in tubes containing sodium fluoride preservative. The concentration of ethanol in urine (UAC) was determined by headspace gas chromatography, and gas chromatography–mass spectrometry and high-performance liquid chromatography were used to analyze 5HTOL and 5HIAA (7–9). In control experiments without the intake of any alcohol, the UAC in the morning was always zero, and the 5HTOL/5HIAA ratio was less than the cutoff concentration of 15 pmol/nmol. (This value was derived from analyzing urine from more than 150 abstaining subjects for whom the 5HTOL/5HIAA ratio was always less than 15 pmol/nmol.) Table I presents the UAC, BAC, and the urinary 5HTOL/5HIAA ratio in samples collected in the morning and afternoon after heavy drinking the night before. The highest UAC in the morning was 171 mg/100 mL, and the corresponding BAC was 35 mg/100 mL. A significantly higher UAC compared with BAC indicates a pooling effect from a previously much higher BAC. Table I shows that the 5HTOL/5HIAA ratios were always abnormally high and remained elevated late into the afternoon, whereas the BAC had already reached zero. This time lag in the 5HTOL/5HIAA ratio compared with BAC and UAC emphasizes the usefulness of this biochemical marker in accident investigations for disclosing recent heavy drinking long after the alcohol has been metabolized.
This study has important implications in forensic and clinical medicine when the likely causes of serious accidents are being investigated by the police. Tangible evidence to suggest that the person or persons involved might have been drinking heavily the night before could prove significant in criminal and civil litigation when responsibility for the accident is assigned and insurance claims are made because of negligence. In the United States, postaccident tests for alcohol are a statutory requirement (Omnibus Transportation Employee Testing Act of 1991). However, a negative blood, breath, or urine test for alcohol does not exclude heavy drinking the previous evening. Measuring the ratio of 5HTOL/5HIAA in urine would prove a useful strategy to test whether a person had been drinking heavily and might be impaired due to the aftereffects of alcohol. This biochemical test has been used previously in connection with rehabilitation of alcoholics and drug abusers required to refrain from drinking alcohol (6,10). The present work suggests that there are applications of the 5HTOL/5HIAA ratio in forensic science as well.

References

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6. A. Helander, O. Beck, and S. Borg. The use of 5-hydroxytryptophol as an alcohol intake marker. Alcohol Alcoholism (Suppl 2): 497–502 (1994).
7. A.W. Jones and J. Schuberth. Computer-aided headspace gas chromatography applied to blood alcohol analysis: importance of on-line process control. J. Forensic Sci. 34: 1116–27 (1989).
8. A. Helander, O. Beck, M. Wennberg, T. Wikström, and G. Jacobsson. Determination of urinary 5-hydroxyindole-3-acetic acid by high-performance liquid chromatography with electrochemical detection and direct sample injection. Anal. Biochem. 196: 170–73 (1991).
9. O. Beck, S. Borg, L. Eriksson, and A. Lundman. 5-Hydroxytryptophol in the cerebrospinal fluid and urine of alcoholics and healthy subjects. Naunyn–Schmiedeberg’s Arch. Pharmacol. 321: 293–97 (1982).
10. A. Voltaire, O. Beck, and S. Borg. Urinary 5-hydroxytryptophol: a possible marker of recent alcohol consumption. Alcoholism Clin. Exp. Res. 16: 281–85 (1992).
    

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