Urinary ethyl glucuronide for the assessment of alcohol consumption during pregnancy: Comparison between biochemical data and screening questionnaires

Background: Ethyl glucuronide (EtG) is a metabolite of ethanol used as a marker of alcohol drinking and is identified in urine. Gestational alcohol drinking harms the fetus, so, disclosing any form of use and abuse of this substance during pregnancy is crucial. Many discovery methods have been planned to overcome this question, including that of using screening questionnaires as the AUDIT-C, T-ACE/TACER-3, and TWEAK. Aim: The aim and novelties of this study were to compare biochemical data from urinary EtG assays (cut-off 100 ng/mL for risking drinking behavior) with the outcome of questionnaires and of a food diary routinely used in our hospital; moreover, for the first time, we analyzed in pregnant women the EtG values normalized by the amount of creatinine excreted according to methods previously established [1]. Methods: Random urine samples were collected from 309 pregnant women immediately after being interviewed. EtG was quantified using an enzyme immunoassay and urinary creatinine was assessed using an enzymatic colorimetric method. Women that had not exhaustively answered one of the questionnaires, or that refused to provide urine samples were excluded. In the end, 309 women had a complete set of data and were considered for this study. Urine creatinine measurements were performed to determine if urine dilution might have resulted in false negatives in the challenge study. In order to accomplish this objective, as urinary creatinine concentrations are, on average, approximately 1 mg/mL, we used a normalized value of 100 ng EtG/mg Creatinine [1]. Results: Our data show that 20.4% of the pregnant women in the study were over the established normalized cut-off value. Poor to null concordance (unweighted k < 0.2) was found between EtG data and the screening interviews, that show, on average, lower levels of alcohol consumption. Conclusion: In conclusion, this study provides evidence that the assessment of maternal alcohol consumption during pregnancy, only indirectly estimated with questionnaires and food diary, can produce misleading ratings.

Determination of cortisol in hair using UHPLC–MS/MS: application to patients admitted for ethanol dependence treatment

Aim: Cortisol hair levels can be used to evaluate chronic stress status. In this context, an improved UHPLC–MS/MS assay for the determination of cortisol in hair was developed and validated. Materials & methods: Hair was extracted with methanol for 4 h at 25°C. Chromatographic run time was 5.5 min. The assay was linear in the range of 1–250 pg mg-1. Precision was 3.6–12.2% and accuracy 97.1–103.8%. The method was applied in hair from 19 volunteers admitted at a rehabilitation clinic, with ethanol consumption classified using ethyl glucuronide hair levels. Conclusion: Abstinent/chronic moderate ethanol consumers had significantly lower cortisol hair levels than chronic excessive consumers. This is the first study evaluating cortisol hair levels in ethanol abuse patients using an objective marker for ethanol consumption.

A simplified approach for determination of urinary ethyl glucuronide by gas chromatography–mass spectrometry

Urinary ethyl glucuronide (EtG), an alcohol biomarker, plays an essential role in monitoring alcohol abstinence and relapse during treatment for alcohol dependence. Detection of this biomarker has become a routine in many clinical and forensic laboratories over the last few years. Most previously published methods commonly use hyphenated chromatographic techniques along with extensive extraction procedure before analysis. This work aimed to develop and validate an electron impact ionization mode gas chromatography–mass spectrometry method to measure ethyl glucuronide levels in human urine. For its determination, urine samples were dried under a gentle stream of nitrogen, derivatized with N,O-bis(trimethylsilyl) trifluoroacetamide, incubated, and injected into the instrument. The analysis was performed using single quadrupole gas chromatography–mass spectrometry (GC-MS) technology and validation was performed according to the guidelines of the German Society of Toxicology and Forensic Chemistry (GTFCh). The linearity of urinary EtG was obtained in the range of 30–5000 ng/ml with a correlation coefficient (r) above 0.999. The extraction recoveries exceeded 80%, and the obtained inter-day and intra-day precisions were below 15%. The achieved limit of detection was 10 ng/ml and limit of quantification achieved was 30 ng/ml. The electron ionization gas chromatography–mass spectrometry technique proves to be a feasible option for determining EtG in human urine when other sophisticated techniques are unapproachable. This method provides a good sensitivity and proves to be cost-effective, robust, and advantageous for both clinical as well as forensic settings.

Driving license regranting in DUI subjects: Road accident variables and predictive factors of substance use disorder

A comparative case study (2017–2020) was conducted to identify demographic, social, medico-legal, and toxicological variables associated with non-fatal accidents in driving under the influence (DUI) subjects. A second aim was to identify the factors predictive of substance use disorders among subjects. Drivers charged with alcohol DUI (blood alcohol concentration (BAC) > 0.5) and/or psychoactive substance DUI were included; cases included those involved in an accident while intoxicated, and the comparison group included DUI offenders negative for road accident involvement. Significance was determined by chi-square and Mann–Whitney tests. To prevent confounding effects, a multivariate binary logistic regression analysis was performed. Our sample encompassed 882 subjects (381 in the case group and 501 in the comparison group). Parameters such as psychoactive substances and BAC at the time of the road crash/DUI and the day of the week, when subjects were involved in the road accident or found DUI, resulted in significant differences ( p < 0.01) between groups. The model’s independent variables of BAC > 1.5 g/L ( p = 0.013), BAC > 2.5 g/L ( p < 0.001), and concurrent alcohol and psychoactive substance use ( p < 0.001) were independent risk factors for an accident. Smoking >20 cigarettes/day was an independent risk factor for unfitness to drive ( p < 0.01). Unfitness to drive was based primarily on ethyl glucuronide levels >30 pg/mg. Our results suggest a detailed assessment of DUI subjects with variables associated with accidents (BAC > 1.5 g/L and concurrent intake of psychoactive substances). Hair analysis, including ethylglucuronide (EtG) concentration, should be always performed. Based on our results, nicotine use should be investigated in cases of driving license regranting.

Child alcohol use disorder in Eastern Uganda: screening, diagnostics, risk factors and management of children drinking alcohol in Uganda (TREAT C-AUD): a mixed-methods research protocol

BackgroundFollowing a finding of alcohol use among children aged 5–8 years old in Mbale, Uganda, this project investigates the magnitude of alcohol and substance use among children ged 6–13 years old and related household, community, school, health system and clinical factors.MethodsThe project includes four larger work packages (WPs). WP1 comprises management, WP2 and 3 include the scientific components and WP4 includes integration of results, dissemination, policy and implementation advice. This protocol presents the planned research work in WP 2 and 3. WP2 comprises the adaptation and validation of the alcohol use screening tool Car-Relax-Alone-Forget-Family and Friends-Trouble (CRAFFT) to the age group and setting. WP3 comprises four substudies (SS). SS1 is a cross-sectional community household survey with an estimated sample size of 3500 children aged 6–13 years and their caregivers. We apply cluster sampling and systematic sampling within the clusters. Data collection includes a structured questionnaire for caregiver and child, measuring social and demographic factors, mental health status, alcohol and substance use, nutrition history and anthropometry. Urine samples from children will be collected to measure ethyl glucuronide (EtG), a biological marker of alcohol intake. Further, facilitators, barriers and response mechanisms in the health system (SS2) and the school system (SS3) is explored with surveys and qualitative assessments. SS4 includes qualitative interviews with children. Analysis will apply descriptive statistics for the primary outcome of establishing the magnitude of alcohol drinking and substance use, and associated factors will be assessed using appropriate regression models. The substudies will be analysed independently, as well as inform each other through mixed methods strategies at the stages of design, analysis, and dissemination.Ethics and disseminationData protection and ethical approvals have been obtained in Uganda and Norway, and referral procedures developed. Dissemination comprises peer-reviewed, open access research papers, policy recommendations and intersectoral dialogues.Trial registration numberClinicaltrials.gov 29.10.2020 (#NCT04743024).