Alcohol Hair Research Articles

Interpretation of a highly positive ethyl glucuronide result together with negative fatty acid ethyl esters result in hair and negative blood results

Considering the widespread nature of alcohol-related problems, the diagnosis of excessive alcohol consumption is an important task from medical and legal viewpoints. Alcohol abuse can be documented by usual blood [e.g., carbohydrate deficient transferrin (CDT)] and liver function [e.g., γ-GT or mean corpuscular volume (MCV)] tests, and, over a long-term basis, by hair analysis. Major markers of ethanol consumption in hair are ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs). Detection of EtG in hair is said to be associated with excessive alcohol consumption, whereas a negative result does not unambiguously exclude alcohol abuse. Investigations on FAEEs can also be used to monitor excessive alcohol consumption. Four FAEEs (ethyl myristate, ethyl palmitate, ethyl oleate, and ethyl stearate) are the most suitable markers for the detection of heavy alcohol consumption and show different concentrations in hair of children, adult teetotalers, and social drinkers in comparison with FAEEs concentrations found in the hair of alcoholics. The Society of Hair Testing has provided guidelines for hair testing for chronic excessive alcohol consumption, and states positive cutoffs for a 0–3 cm hair segment as 30 pg/mg and 0.5 ng/mg for EtG and FAEEs, respectively. This study reviews the difficulties in interpreting blood and hair results of a 41-year-old woman involved in a child custody legal dispute. Her EtG and FAEEs concentrations in a 0–3 cm segment were 203 pg/mg and 0.29 ng/mg, respectively, and blood parameters were in the normal range (0.6 %, 93 fl, 14 IU/l for CDT, MCV, and γ-GT, respectively). Although the high EtG concentration suggested excessive drinking behavior, the second hair marker, FAEEs, and the three bloods tests were inconspicuous and in accordance with the claim of abstinence from alcohol by the subject. The woman declared having dyed her hair more than 6 months prior to sampling, use of weight-loss medication, and consumption of about four energy drinks per day. A hair lotion based on ethanolic plant extracts was regularly used by the subject, and could have been a source of contamination. Therefore, we recommend that possible external sources of hair contamination always be taken into consideration, especially if contradictory biological results are obtained and if the subject denies any alcohol intake.

Analysis of Fatty Acid Ethyl Esters in Hair by Headspace Solid-Phase Microextraction (HS-SPME) and Gas Chromatography-Mass Spectrometry (GC-MS)

Fatty acid ethyl esters (FAEE) can be used as alcohol markers in hair. The purpose of this study was to validate a new method for the detection of FAEE in hair, reducing the extraction time. Hair samples were analyzed by gas chromatography-mass spectrometry after headspace solid-phase microextraction with deuterated internal standards. Linear curves for ethyl myristate, palmitate and stearate were obtained in the range considered. The limits of detection were 0.8, 0.005, and 0.01 and the lower limits of quantification were 1.8, 0.01, and 0.4 for ethyl myristate, palmitate and stearate, respectively. Intra- and inter-assay precision and accuracy were less than 12.5% and 16.1%, respectively. Recoveries were higher than 13.5% in all cases. Finally, the method was applied to analyze hair of alcoholics, social drinkers, and teetotallers.

The Reliability of Fatty Acid Ethyl Esters (FAEE) as Biological Markers for the Diagnosis of Alcohol Abuse

The search for biochemical markers for the objective diagnosis of alcoholism has been a topic of research because of the important clinical and forensic implications. In the last few years, two minor ethanol metabolites (ethylglucuronide and fatty acid ethyl esters) have been mainly investigated in hair samples for their ability to be incorporated into this biological matrix. The aim of this study was to experience the detection of fatty acid ethyl esters (FAEE) in the hair of alcoholics, social drinkers, and teetotallers in order to give a contribution to the existing literature. Hair samples from 12 alcoholics, 10 social drinkers, and 10 teetotallers were analyzed by gas chromatography-mass spectrometry technique after headspace solid-phase microextraction with deuterated internal standards. A slight overlap in FAEE concentration between the three groups was found, probably because of external contamination. This observation suggests particular attention to the interpretation of the results. Nevertheless, the results obtained show the usefulness of these biochemical markers in the diagnosis of alcoholism.

Comparison of ethyl glucuronide and fatty acid ethyl ester concentrations in hair of alcoholics, social drinkers and teetotallers

In previous investigations hair analysis for ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEE) proved to be suitable for the detection of excessive alcohol consumption. The aim of this study was to compare EtG and FAEE concentrations in hair of alcoholics, social drinkers and teetotallers. Hair samples from 10 alcoholics in withdrawal treatment, 11 fatalities with documented excessive alcohol consumption, four moderate social drinkers who consumed up to 20 g ethanol per day, and three strict teetotallers were analysed. After external degreasing with n-heptane, extraction with a dimethyl sulfoxide/ n-heptane mixture and headspace solid-phase microextraction of the extracts, four fatty acid ethyl esters (FAEEs) (ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate) were analysed by gas chromatography–mass spectrometry (GC–MS) with deuterated internal standards. EtG was determined by GC–MS/NCI after ultrasonication of the samples with H 2O, cleanup by SPE with aminopropyl columns and PFP derivatisation. The following concentrations were measured for the four groups: teetotallers EtG < 0.002 ng/mg, FAEE 0.05–0.37 ng/mg, moderate social drinkers EtG < 0.002 ng/mg, FAEE 0.26–0.50 ng/mg, alcoholic patients EtG 0.030–0.415 ng/mg, FAEE 0.65–20.50 ng/mg and the fatalities with alcohol history EtG 0.072–3.380 ng/mg, FAEE 1.30–30.60 ng/mg. The results confirm that by using a cut-off value of the sum of FAEE > 1 ng/mg and/or a positive EtG result in hair, excessive alcohol consumption can be identified using hair analysis. However, no significant correlation between the EtG and FAEE concentrations in the positive cases could be shown. Segmental analysis of some of the specimens did not reveal the same distribution for EtG compared to FAEE in hair, and no chronological accordance compared to the self-reported alcohol consumption could be observed for both parameters. These different results of both methods are discussed in terms of differences between EtG and FAEE in mechanism of formation and incorporation into hair and elimination from hair.

Are there possibilities for the detection of chronically elevated alcohol consumption by hair analysis? A report about the state of investigation

The analysis of suitable ethanol markers in hair would be an advantageous tool for chronic alcohol abuse control because of the wide diagnostic window allowed by this specimen and the possibility of segmental investigation. Between the markers practically used or thoroughly investigated in blood or urine, ethylglucuronide, fatty acid ethylesters, phosphatidylethanol, acetaldehyde adducts to protein and 5-hydroxytryptophol can be regarded as possible candidates also in hair, but preliminary data were found in the literature only for ethylglucuronide and acetaldehyde modified proteins.By using headspace gas chromatography and headspace solid phase microextraction in combination with gas chromatography–mass spectrometry (SPME-GC/MS), in alkaline hydrolysates of hair it was possible to determine between 17 and 135 ng/mg of ethanol beside acetone and several other volatile compounds with slightly higher ethanol values for alcoholics than for social drinkers and teetotalers. A part of this is ethanol only absorbed in the hair matrix from the surrounding environment and consequently is not applicable as a diagnostic criterion. By extraction with aqueous buffer, methanol or a methanol/chloroform mixture and subsequent alkaline hydrolysis it was found that another part is generated from ethylesters, which are preferentially deposited in the lipid fraction of hair. In a specific search for ethylesters of 17 carboxylic acids by GC/MS-SIM in most cases ethyl 4-hydroxybenzoate (0.1 to 5.9 ng/mg, a preservative in hair cosmetics) and in four cases traces of indolylacetic acid ethylester were found. Furthermore, diethyl phthalate (a softening agent, present also in many cosmetic products) was identified in the hair of alcoholics as well as of children. As potential markers of alcohol intake, ethyl palmitate, ethyl stearate and ethyl oleate were detected in hair samples of alcoholics by headspace SPME-GC/MS of the chloroform/methanol extracts.