Ethyl glucuronide in hair detects a high rate of harmful alcohol consumption in presumed non-alcoholic fatty liver disease.

BACKGROUND & AIMS: Non-alcoholic fatty liver disease (NAFLD) and alcohol-related liver disease (ALD) cannot reliably be distinguished by routine diagnostics, and the role of alcohol consumption in metabolic dysfunction-associated fatty liver disease (MAFLD) remains unclear. We investigated alcohol consumption in patients with presumed NAFLD and ALD using novel objective alcohol markers. METHODS: In total, 184 consecutive patients were included in this prospective observational study. Alcohol intake was assessed by ethylglucuronide in hair (hEtG) and urine (uEtG); the utility of these measures for alcohol detection was compared to Alcohol Use Disorders Identification Test-Consumption (AUDIT-C), carbohydrate deficient transferrin (CDT), mean corpuscular volume (MCV), gamma-glutamyltransferase (GGT), and ALD/NAFLD index (ANI). Clinical characteristics of patients with NAFLD and ALD were re-assessed after reclassification based on repeated moderate (≥10 g <60 g EtOH/day) and excessive (≥60 g EtOH/day) alcohol consumption, and patients were retrospectively reclassified based on MAFLD criteria. RESULTS: Repeated moderate to excessive alcohol consumption was detected in 28.6%, 28.5%, and 25.0% of patients with presumed NAFLD, ALD or MAFLD, respectively. ANI score, AUDIT-C, uEtG, and hEtG showed AUCs of 0.628, 0.733, 0.754, and 0.927 for the detection of repeated moderate to excessive alcohol consumption, respectively. The indirect markers CDT, MCV and GGT were not reliable. Patients with repeated moderate or excessive alcohol consumption were significantly more often male, had a significantly lower BMI, and suffered significantly less often from type 2 diabetes or impaired glucose tolerance. CONCLUSIONS: In total, 28.6% of patients with presumed NAFLD, and 25.0% with MAFLD are at risk of alcohol-related liver damage. AUDIT-C, uEtG and hEtG should be used to screen for alcohol consumption in patients with fatty liver disease. LAY SUMMARY: Fatty liver disease can be caused by metabolic factors and/or alcohol consumption. The diagnosis of non-alcoholic fatty liver disease (NAFLD) is based on the exclusion of harmful alcohol consumption, while metabolic dysfunction-associated fatty liver disease (MAFLD), which has been proposed as a new name for NAFLD, is based on the presence of metabolic comorbidities and allows for alcohol consumption. Herein, we show that up to 29% of patients diagnosed with NAFLD and 25% with MAFLD are at risk of alcohol-related liver damage. We show that ethyl glucuronide (a metabolite of alcohol) in the hair and urine can accurately detect potentially harmful alcohol consumption in these patients - as such, these tests should be integrated into routine diagnostic work-up for patients with fatty liver disease.

Detox shampoos for EtG and FAEE in hair - Results from in vitro experiments.

The assessment of alcohol consumption behavior in hair is well established in forensic toxicology. The Society of Hair Testing (SoHT) recommends the direct alcohol markers ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEE) for the detection of past alcohol consumption. In this study, we investigated if detox shampoos which are sold online can have an impact on EtG or FAEE concentrations in hair. According to customer reviews, positive drug testing results could be avoided after long-term incubation with shampoo under a swimming cap. To evaluate the potential of four detox shampoos, we incubated distal hair samples from three subjects, obtained during a standard haircut, for 2.5, 5, 7.5, and 10 hours. For EtG, three shampoos performed similar to deionized water. The fourth shampoo showed additional heavy washout effects with a decrease of up to 86% after 2.5 hours. For the apolar FAEE, no washout was observed. Incubation with two shampoos resulted in increases in FAEE concentrations due to FAEE being present as shampoo ingredients. Further investigation of EtG washout in proximal forensic hair samples (n = 9) with the most potent shampoo showed a mean decrease in deionized water of 23% ± 25%, and a decrease by the use of detox shampoo of 73% ± 12%, compared to non-incubated hair after 8 hours. In conclusion, detox shampoos proved to have the potential to alter EtG and FAEE concentrations in hair during in vitro experiments.

Commentary on current changes of the SoHT 2016 consensus on alcohol markers in hair and further background information.

The consensus on alcohol markers in hair was revised for the fourth time by an expert group of the Society of Hair Testing based on current state of research. This revision was adopted by the members of the Society during the business meeting in Brisbane on August 29th 2016. For both markers, ethyl glucuronide (EtG) and fatty acid ethyl esters (FAEEs), two cut-off values for discrimination between teetotalers or occasional low amount consumption and moderate alcohol drinking (low cut-off), and between non-excessive (abstinence up to moderate alcohol intake) and chronic excessive drinking (high cut-off value) were critically examined. For the current revision, the cut-off values for EtG (7pg/mg and 30pg/mg, respectively) remained unchanged despite different findings or discussions published in the meantime. This was mainly due to the lack of broader data collections from new studies with great numbers of volunteers following thorough study concepts. In contrast, an essential change of the consensus was accepted for the FAEEs, where the concentration of ethyl palmitate (E16:0) can be used autonomously for interpretation instead of the concentration sum (ΣFAEE) of the four esters ethyl myristate, ethyl palmitate, ethyl oleate and ethyl stearate, as previously applied. After evaluation of the data from seven laboratories, the E16:0 cut-off for abstinence assessment was defined at 0.12ng/mg for the 0-3cm segment and at 0.15ng/mg for the 0-6cm segment. The cut-off for chronic excessive drinking was fixed at 0.35ng/mg for the 0-3cm segment and at 0.45ng/mg for the 0-6cm segment. The use of E16:0 with these cut-offs in place of ΣFAEE for alcohol intake assessment produces only a minor loss in discrimination power, leads to no essential difference in the interpretation concerning chronic excessive alcohol consumption and is suitable to confirm EtG results in abstinence assessment if ethanol containing hair sprays or lotions are excluded.

Effect of the analyzed hair length on fatty acid ethyl ester (FAEE) concentrations in hair--is there congruence of cut-offs for 0-3 and 0-6 cm hair segments?

BACKGROUND: According to the current SoHT consensus for the use of alcohol markers in hair FAEEs can be analyzed in the proximal 0-3 or 0-6 cm segment with the cut-offs 0.2 and 0.4 ng/mg for abstinence assessment and 0.5 and 1.0 ng/mg for chronic excessive alcohol consumption. Both sets of cut-offs should be congruent for uniform interpretation. METHODS: FAEEs were determined in parallel for the 0-3 and the 0-6 cm segment of 157 hair samples and the concentration ratio between both segments (0-3 cm)/(0-6 cm) was calculated. For comparison, EtG was measured in the 0-3 cm segment of 135 of these samples and the FAEE concentration ratio was calculated by re-evaluation of segmental concentrations from further 42 samples of a previous study. RESULTS: The concentration ratio ranged from 0.3 to 1.5 (mean 0.83, median 0.82) and showed that the current cut-offs (ratio 0.50) are not congruent and that the cut-offs of the 0-3 cm are more restrictive against alcohol use or abuse. There was no correlation between the ratio and the concentration of FAEEs or EtG showing that the ratio does not depend on the drinking amount. Furthermore, no significant difference of the ratio was found between cosmetically untreated, bleached, dyed or spayed hair samples. Adaptation of the 0-3 cm cut-offs by increasing to 0.3 and 0.8 ng/mg respectively improved the congruence of both segment lengths but did not lead to a better agreement between FAEE and EtG interpretation. CONCLUSIONS: It follows from the results and further literature data that the variable length of the proximal hair segment between 3 and 6 cm which is possible according to the present SoHT consensus additionally contributes to the uncertainty of interpretation caused by biological variability and hair cosmetics. Furthermore, it is improbable from the present state of knowledge that a proximal segment length above 3 cm unambiguously enables the detection of alcohol consumption or abuse occurring more than 3 months before sampling. Therefore, it should be considered in a future consensus to redefine the hair length for both markers on 0-3 cm.