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Background: Alcohol abuse is one of the most common causes of mortality worldwide. This study aimed to investigate the efficacy of a treatment in reducing circulating ethanol and oxidative stress biomarkers. Methods: Twenty wine-drinking subjects were investigated in a randomized controlled, single-blind trial (ClinicalTrials.gov. Identifier: NCT06548503; Ethical Committee of the University of Padova (HEC-DSB/12-2023) to evaluate the effect of the intake of a product containing silymarin, pyrroloquinoline quinone sodium salt, and myricetin (referred to as Si.Pi.Mi. for this project) on blood alcohol, ethyl glucuronide (EtG: marker for alcohol consumption) and markers of oxidative stress levels (Reactive Oxygen Species—ROS, Total Antioxidant Capacity—TAC, CoQ10, thiols redox status, 8-isoprostane, NO metabolites, neopterin, and uric acid). The effects of the treatment versus placebo were evaluated acutely and after 1 week of supplementation in blood and/or saliva and urine samples. Results: Si.Pi.Mi intake reduced circulating ethanol after 120 min (−33%). Changes in oxidative stress biomarkers, particularly a TAC (range +9–12%) increase and an 8-isoprostane (marker of lipidic peroxidation) decrease (range −22–27%), were observed too. Conclusion: After the administration of Si.Pi.Mi, the data seemed to suggest a better alcohol metabolism and oxidative balance in response to wine intake. Further verification is requested. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the alcohol consumption among professional truck and bus drivers using direct ethanol biomarkers, and to explore its relationship with anxiety, depression, and stress. The assessment of potential harmful drinking was conducted through the measurement of direct biomarkers: phosphatidylethanol (PEth), ethyl glucuronide (EtG), and ethyl sulfate (EtS), using dried blood spots (DBS). Additionally, self-reported data from the Alcohol Use Disorders Identification Test (AUDIT-C) were used. Emotional states, including depression, anxiety, and stress, were evaluated using the Depression, Anxiety, and Stress Scale (DASS-21). A total of 97 drivers participated in the study, with the majority being male (96%) and identified as truck drivers (75.3%). Among them, 43.3% reported working more than 10 h daily. The majority of volunteers exhibited normal levels of stress (81.4%), anxiety (83%), and depression (86.6%). According to the AUDIT-C assessment, 30.9% were categorized as having a moderate risk, while 11.3% were deemed to be at high risk for harmful alcohol consumption behavior. Ethyl glucuronide (EtG) and ethyl sulfate (EtS) levels, indicating recent ethanol consumption, were detected in 14.4% of the drivers. In contrast, the long half-life metabolite PEth (16:0-18:1) was present in 88.7% of the volunteers. A moderate correlation (rs = 0.45, p <.01) was observed between PEth levels and AUDIT-C scores. The Receiver Operating Characteristic (ROC) curve, utilizing a PEth threshold of ≥ 59.0 ng ml−1, displayed 78% sensitivity and 73% specificity in effectively distinguishing high risk for alcohol intake. Notably, no significant associations were found between alcohol consumption and levels of stress, depression, and anxiety. The study findings indicate a noteworthy proportion of drivers engaging in regular alcohol consumption alongside a demanding workload. Notably, PEth measurements highlighted an underreporting within the AUDIT-C self-reports. These results lend robust support for the utilization of biomarkers in assessing alcohol consumption patterns among drivers. [ABSTRACT FROM AUTHOR]

This article traces the historical development of various biomarkers of acute and/or chronic alcohol consumption. Much of the research in this domain of clinical and laboratory medicine arose from clinics and laboratories in Sweden, as exemplified by carbohydrate deficient transferrin (CDT) and phosphatidylethanol (PEth). Extensive studies of other alcohol biomarkers, such as ethyl glucuronide (EtG), ethyl sulfate (EtS), and 5‐hydroxytryptophol (5‐HTOL), also derive from Sweden. The most obvious test of recent drinking is identification of ethanol in a sample of the person's blood, breath, or urine. However, because of continuous metabolism in the liver, ethanol is eliminated from the blood at a rate of 0.15 g/L/h (range 0.1–0.3 g/L/h), so obtaining positive results is not always possible. The widow of detection is increased by analysis of ethanol's non‐oxidative metabolites (EtG and EtS), which are more slowly eliminated from the bloodstream. Likewise, an elevated ratio of serotonin metabolites in urine (5‐HTOL/5‐HIAA) can help to disclose recent drinking after ethanol is no longer measurable in body fluids. A highly specific biomarker of hazardous drinking is CDT, a serum glycoprotein (transferrin), with a deficiency in its N‐linked glycosylation. Another widely acclaimed biomarker is PEth, an abnormal phospholipid synthesized in cell membranes when people drink excessively, having a long elimination half‐life (median ~6 days) during abstinence. Research on the subject of alcohol biomarkers has increased appreciably and is now an important area of drug testing and analysis. [ABSTRACT FROM AUTHOR]

Monitoring long‐term alcohol use and/or abstinence is essential in clinical and medico‐legal cases. Analysis of ethyl glucuronide (EtG) in hair provides information on alcohol consumption over several months. However, there is a lag time between ethanol consumption, incorporation of EtG in the hair bulb and hair growing out of the scalp. Phosphatidylethanol (PEth) 16:0/18:1 analysis in whole blood has a detection window of 2–4 weeks, allowing for the detection of recent alcohol consumption. In this study, 2340 paired samples (of hair and venous whole blood from 1170 individuals) were analysed for EtG in hair (hEtG) and PEth 16:0/18:1 in venous whole blood. PEth 16:0/18:1 and hEtG results were subdivided into three categories according to the consensus of SoHT (hEtG) and PEth‐NET (PEth): abstinence/low, moderate or excessive alcohol consumption. For hEtG analysis, 446 individuals presented abstinence/low alcohol consumption, of which 2% were classified as excessive alcohol users through PEth 16:0/18:1 analysis. This suggests excessive alcohol consumption in the weeks before sample collection. Out of 483 individuals classified as heavy alcohol users based on hEtG analysis, 14% showed abstinence/low alcohol consumption for PEth 16:0/18:1 analysis, implying that these subjects stopped drinking 2–4 weeks before sample collection. Our results show that the analysis of the two different biomarkers can lead to a more accurate categorisation of individuals. Therefore, we emphasize that for the retrospective investigation of alcohol use, it is necessary to include two alcohol use biomarkers with different detection windows. [ABSTRACT FROM AUTHOR]

Alcohol-associated liver disease has seen a significant rise in the last 2 decades, with an associated rise in the need for accurate alcohol use assessment. Alcohol use has been associated with poor outcomes in both the pre-liver transplant and post-liver transplant patients. Patients with alcohol use disorder often under-report their alcohol consumption because of varying factors, highlighting the need for objective assessment of alcohol use. Aside from the available self-report questionnaires, multiple serologic biomarkers are currently available to assist clinicians to assess recent alcohol consumption among patients with chronic liver disease, liver transplant candidates, and recipients. In this review, we will assess some of these alcohol biomarkers, discuss their strengths and weakness, and review-available data to discuss their role in pre-liver transplant and post-liver transplant population.

Abstract Alcohol and cocaine (COC) are commonly co-used drugs that cause addiction and have harmful effects. Their abuse may threaten the health of the abuser and public safety by causing serious accidents or crimes. The recidivism rate of drug-related crimes closely correlates with alcoholism. Several incidences of alcohol consumption in combination with drug abuse have been reported. Here, liquid chromatography tandem mass spectrometric method was developed to simultaneously analyze ethyl glucuronide (EtG), a metabolite of ethanol; COC; cocaethylene (CE), an alcohol-derived metabolite of COC; and benzoylecgonine (BZE), a major metabolite of COC, to determine the concurrent use of alcohol with COC. For pre-treatment, ultracentrifugation (5 min, 50,000 g) and mixed-mode anion exchange solid-phase extraction were used to increase the recovery of target compounds and minimize the matrix effect of hair. The lower limits of quantification were: 7 pg/mg (EtG), 2 pg/mg (COC), 10 pg/mg (CE), and 1 pg/mg (BZE). The correlation coefficient (r) of the calibration curve within the quantified range of target compounds was ≥ 0.9978. The intra- and inter-day accuracies were − 6.1–9.7% and − 9.3–8.3%, and intra- and inter-day precisions were 0.5–10.3% and 0.6–14.4%, respectively. The recovery, matrix effect, process efficiency, and autosampler stability were 89.2–104.8%, 81.6–105.4%, 81.5–107.1%, and 96.6–109.7%, respectively. The novel analytical method was validated with hair samples from individuals suspected of alcohol and COC use, and the method could distinguish between independent and concurrent use. Based on the findings, the analytical approach developed in this study is anticipated to be valuable in drug and alcohol dependence tests that require the simultaneous detection of alcohol and COC abuse.

Alcohol and cocaine (COC) are commonly co-used drugs that cause addiction and have harmful effects. Their abuse may threaten the health of the abuser and public safety by causing serious accidents or crimes. The recidivism rate of drug-related crimes closely correlates with alcoholism. Several incidences of alcohol consumption in combination with drug abuse have been reported. Here, liquid chromatography tandem mass spectrometric method was developed to simultaneously analyze ethyl glucuronide (EtG), a metabolite of ethanol; COC; cocaethylene (CE), an alcohol-derived metabolite of COC; and benzoylecgonine (BZE), a major metabolite of COC, to determine the concurrent use of alcohol with COC. For pre-treatment, ultracentrifugation (5 min, 50,000 g) and mixed-mode anion exchange solid-phase extraction were used to increase the recovery of target compounds and minimize the matrix effect of hair. The lower limits of quantification were: 7 pg/mg (EtG), 2 pg/mg (COC), 10 pg/mg (CE), and 1 pg/mg (BZE). The correlation coefficient (r) of the calibration curve within the quantified range of target compounds was ≥ 0.9978. The intra- and inter-day accuracies were − 6.1–9.7% and − 9.3–8.3%, and intra- and inter-day precisions were 0.5–10.3% and 0.6–14.4%, respectively. The recovery, matrix effect, process efficiency, and autosampler stability were 89.2–104.8%, 81.6–105.4%, 81.5–107.1%, and 96.6–109.7%, respectively. The novel analytical method was validated with hair samples from individuals suspected of alcohol and COC use, and the method could distinguish between independent and concurrent use. Based on the findings, the analytical approach developed in this study is anticipated to be valuable in drug and alcohol dependence tests that require the simultaneous detection of alcohol and COC abuse. [ABSTRACT FROM AUTHOR]

Ethanol exposure during pregnancy is an important problem and is the cause of fetal alcohol syndrome (FAS) and fetal alcohol spectrum disorder (FASD). The etiology of FAS and FASD can be elucidated using animal models. Recently, a novel model, the zebrafish (Danio rerio), has garnered the interest of researchers. This study confirmed the negative influence of ethyl alcohol (0.5 %, 1.5 %, and 2.5 % v/v) on the development of zebrafish embryos. The observed malformations included pericardial and yolk sac edema, increased body curvature, tail edema, and a decreased embryo hatching rate. The differences in body length, body width, and heart rate were statistically significant. Due to the similarities in the quantity and function of ethanol biotransformation enzymes between zebrafish and mammals, this study investigated the nonoxidative metabolites of ethanol – ethyl glucuronide (EtG) and ethyl sulfate (EtS) – in zebrafish following ethanol exposure. This research confirmed that EtG and EtS concentrations can be measured in zebrafish embryos, and the levels of these metabolites appear to be associated with the ethyl alcohol concentration in the medium. • The toxic effect of ethanol on zebrafish embryos has been confirmed. • Non-oxidative metabolites have been observed following the exposure of embryos to ethanol. • EtG and EtS amount in zebrafish embryos seemed to be associated with ethanol concentration. [ABSTRACT FROM AUTHOR]

Copyright of Ege Journal of Medicine is the property of Ege University, Faculty of Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Marc Bartel,1,&ast; Vanessa Hofmann,1,&ast; Shijin Wang,2,3 Johannes Mueller,2 Tom R Sundermann,1 Sebastian Mueller2 1Institute of Forensic and Traffic Medicine, Heidelberg University Hospital, Heidelberg, Germany; 2Center for Alcohol Research, University Hospital Heidelberg, Heidelberg, Germany; 3Department of Gastroenterology, The Affiliated Hospital of Qingdao University, Shandong, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Sebastian Mueller, Center for Alcohol Research, University Hospital Heidelberg, Im Neuenheimer Feld 350, Heidelberg, 69120, Germany, Tel +49 (0) 6221 567121, Email sebastian.mueller@urz.uni-heidelberg.dePurpose: Ethyl glucuronide (EtG), ethyl sulfate (EtS) and phosphatidylethanol (PEth) are considered specific direct biomarkers for detecting alcohol consumption. However, PEth, which is produced in red blood cells (RBC), varies considerably between patients for unknown reasons. We here studied various confounders of PEth elimination including fibrosis after alcohol withdrawal.Patients and Methods: EtG, EtS and PEth together with routine laboratory and clinical parameters were studied in 100 Caucasian heavy drinkers prior and after alcohol detoxification. In addition, fibrosis stage and degree of steatosis were assessed by transient elastography (Fibroscan, Echosens, Paris).Results: All three biomarkers were highly correlated (0.61– 0.72) with initial serum alcohol levels, but only PEth correlated with daily alcohol consumption. After alcohol withdrawal, PEth significantly decreased within 6.1 days from 1708 to 810 ng/mL (half-life varied from 1.6 to 15.2 days). Both levels of serum alcohol but also EtG and EtS were higher in patients with liver cirrhosis as compared to patients without fibrosis despite comparable alcohol consumption suggesting a decreased alcohol elimination in patients with cirrhosis. PEth was also elevated in cirrhosis but not significantly. In contrast, PEth elimination rate was significantly higher in patients with enhanced RBC turnover and signs of alcohol-mediated hemolytic anemia with elevated ferritin, LDH and increased mean corpuscular volume (MCV).Conclusion: We here demonstrate that alcohol elimination is decreased in patients with liver cirrhosis. In patients with cirrhosis, PEth levels are both affected in opposite directions by enhanced red blood cell turnover and elevated alcohol levels. Our data have important implications for the use and interpretation of PEth in the clinical setting.Keywords: ethyl glucuronide, ethyl sulfate, alcohol-related liver disease, liver fibrosis, liver stiffness, alcohol consumption

Background: Alcohol abuse is one of the most common causes of mortality worldwide. This study aimed to investigate the efficacy of a treatment in reducing circulating ethanol and oxidative stress biomarkers. Methods: Twenty wine-drinking subjects were investigated in a randomized controlled, single-blind trial (ClinicalTrials.gov. Identifier: NCT06548503; Ethical Committee of the University of Padova (HEC-DSB/12-2023) to evaluate the effect of the intake of a product containing silymarin, pyrroloquinoline quinone sodium salt, and myricetin (referred to as Si.Pi.Mi. for this project) on blood alcohol, ethyl glucuronide (EtG: marker for alcohol consumption) and markers of oxidative stress levels (Reactive Oxygen Species—ROS, Total Antioxidant Capacity—TAC, CoQ10, thiols redox status, 8-isoprostane, NO metabolites, neopterin, and uric acid). The effects of the treatment versus placebo were evaluated acutely and after 1 week of supplementation in blood and/or saliva and urine samples. Results: Si.Pi.Mi intake reduced circulating ethanol after 120 min (−33%). Changes in oxidative stress biomarkers, particularly a TAC (range +9–12%) increase and an 8-isoprostane (marker of lipidic peroxidation) decrease (range −22–27%), were observed too. Conclusion: After the administration of Si.Pi.Mi, the data seemed to suggest a better alcohol metabolism and oxidative balance in response to wine intake. Further verification is requested.

Problematic alcohol use is prevalent in Russia and is deleterious for individuals with HIV and Hepatitis C Virus (HCV). Ethyl glucuronide (EtG) and blood alcohol content (BAC) provide objective biomarkers of drinking that can be compared to self-reported alcohol use. This paper describes patterns of alcohol use measured by biomarkers and self-report along with concordance across measures. Participants were Russian women with HIV and HCV co-infection (N = 200; Mean age = 34.9) from two Saint Petersburg comprehensive HIV care centers enrolled in an alcohol reduction intervention clinical trial. Measures were: (a) urine specimen analyzed for EtG; (b) breathalyzer reading of BAC; and (c) self-reported frequency of drinking, typical number of drinks consumed, and number of standard drinks consumed in the past month. At baseline, 64.0% (n = 128) had a positive EtG (> 500 ng/mL) and 76.5% (n = 153) had a positive breathalyzer reading (non-zero reading). There was agreement between EtG and BAC (kappa = 0.66, p <.001; Phi coefficient = 0.69, p <.001); self-reported alcohol measures were positively correlated with positive EtG and BAC (p's < 0.001). There was concordance between EtG and BAC measures, which have differing alcohol detection windows. Most participants endorsed frequent drinking at high quantities, with very few reporting no alcohol consumption in the past month. Concordance between biomarkers and self-reported alcohol use suggests that underreporting of alcohol use was minimal. Results highlight the need for alcohol screening within HIV care. Implications for alcohol assessment within research and clinical contexts are discussed. [ABSTRACT FROM AUTHOR]

Aims Phosphatidylethanol (PEth) is only formed when ethanol is present in blood. This direct alcohol marker has been widely discussed, including the minimum amount of ethanol being necessary to form as much PEth as to exceed the threshold of 20 ng/mL in previously PEth negative subjects. In order to corroborate hitherto existing results, a drinking study including 18 participants after a 3-week alcohol abstinence was performed. Methods They consumed a pre-calculated amount of ethanol to reach a blood alcohol concentration (BAC) of at least 0.6 g/kg. Blood was drawn before and periodically seven times after alcohol administration on day 1. Blood and urine were also collected the next morning. Dried blood spots (DBS) were prepared immediately from collected venous blood. BAC was determined by head space gas chromatography and the concentrations of both PEth (16:0/18:1, 16:0/18:2 and five additional homologues) and ethyl glucuronide (EtG) were analysed using liquid chromatography–tandem mass spectrometry. Results Out of 18, 5 participants had concentrations of PEth 16:0/18:1 above the threshold of 20 ng/mL, and 11 out of the 18 subjects had concentrations between 10 and 20 ng/mL. In addition, four persons had PEth 16:0/18:2 concentrations above 20 ng/mL the following morning. All test subjects tested positive for EtG in DBS (≥ 3 ng/mL) and urine (≥100 ng/mL) upon 20–21 h after alcohol administration. Conclusion By combining both a lower cutoff of 10 ng/mL and the homologue PEth 16:0/18:2, the sensitivity to detect a single alcohol intake after a 3-week abstinence increases to 72.2%. [ABSTRACT FROM AUTHOR]

Abstract Background An increasing prevalence of alcohol consumption is a major public health problem, which has also led to an increasing number of children who have been prenatally exposed to the toxic effects of ethanol. However, obtaining reliable information on prenatal alcohol exposure through maternal self-reports has proved difficult. Aims Our aim was to evaluate the potential for rapid screening test for measuring ethyl glucuronide (EtG), a specific alcohol metabolite, from urine samples of pregnant women. Methods Five hundred five urine samples of pregnant women were collected anonymously from five prenatal units in two Finnish cities: a tertiary specialist antenatal clinic for pregnant women with problematic substance use (HAL), a regular hospital antenatal clinic (LCH = Lahti Central Hospital), a prenatal screening unit and two community maternity clinics (USR = user self-recruiting units). All samples were screened using rapid EtG test strips, and all positive, uncertain, and randomly selected negative samples were confirmed by quantitative analyses. The samples were also screened for cotinine and use of cannabis. Results In this material an EtG cut-off of 300 ng/mL suggesting heavy alcohol drinking was exceeded by 7.4% (5/68) of the samples in the HAL clinic, 1.9% (4/202) in LCH, and 0.9% (2/225) in USR. A cut-off of 100 ng/mL was exceeded by 17.6% (12/68) of samples from HAL, 7.5% (16/212) from LCH, and 6.7% (15/225) from USR. Based on confirmatory quantitative analyses, there were no false negatives nor false positives in rapid EtG screening. However, 57 (11.3%) of test results were classified as uncertain. In these cases, confirmation by quantitative analyses resulted in 56.1% rate of positive values. 73% of the samples with EtG > 300 ng/mL showed positive cotinine results suggesting smoking co-occurring with alcohol intake. Conclusions Rapid EtG tests may be an easy and inexpensive method, which may improve the possibilities for screening alcohol use among pregnant women during routine prenatal visits. Quantitative EtG analyses are recommended to confirm screening positive and uncertain cases. Trial registration NCT04571463 Date of Registration 11/05/2020.

Introduction: Detection of alcohol (ETOH) use with biomarkers provides an opportunity to intervene and treat patients with alcohol use disorder before and after liver transplant (LT). We describe our center's experience using urine ethyl glucuronide (EtG) and serum phosphatidylethanol (PEth) in alcohol screening protocols. Methods: Single‐center, retrospective review of patients presenting for LT evaluation, patients waitlisted for LT for alcohol‐associated liver disease (ALD), and patients who received a LT for ALD over a 12‐month period, from October 1, 2019 through September 30, 2020. Patients were followed from waitlisting to LT, or for up to 12 months post‐LT. We monitored protocol adherence to screening for ETOH use‐ defined as completion of all possible tests over the follow‐up period‐ at the initial LT visit, while on the LT waitlist and after LT. Results: During the study period, 227 patients were evaluated for LT (median age 57 years, 58% male, 78% white, 54.2% ALD). Thirty‐one patients with ALD were placed on the waitlist, and 38 patients underwent LT for ALD during this time period. Protocolized adherence to screening for alcohol use was higher for PEth for all LT evaluation patients (191 [84.1%] vs. 146 [67%] eligible patients, p <.001), in patients with ALD waitlisted for LT (22 [71%] vs. 14 (48%] eligible patients, p =.04) and after LT for ALD, 20 (33 [86.8%] vs. 20 [52.6%] eligible patients, p <.01). Few patients with a positive test in any group completed chemical dependency treatment. Conclusions: When screening for ETOH use in pre‐ and post‐LT patients, protocol adherence is higher using PEth compared to EtG. While protocolized biomarker screening can detect recurrent ETOH use in this population, engagement of patients into chemical dependency treatment remains challenging. [ABSTRACT FROM AUTHOR]

Larvae and insects are an important and alternative biological matrix in the development anpost-mortem forensic toxicology. They are very useful when conventional matrices are not available, in particular when the loss of biological fluids, due to the decomposition of corpses or to a traumatic death, occurs. The purpose of this study is to develop and validate an analytical method in Ultra High-Performance Liquid Chromatography at High Resolution Mass Spectrometry (HPLC/HR-MS) for the research and quantification of Ethyl glucuronide (EtG) on larvae. The criteria taken into consideration for the validation are linearity, quantitation limits (LOD and LLOQ), accuracy, precision, carryover, interferences and ionization suppression/enhancement. The method was shown to be linear within the tested range, with a coefficient of determination higher than 0.99. LOD was 2 pg mg−1, while LLOQ was 5 pg mg-1. Accuracy, precision and ionization/suppression enhancement fulfilled the criteria indicated in the guidelines used for the validation. The establishment and validation of this method allowed the identification of Ethyl glucuronide on the larvae of Lucilia sericata (Calliphoridae) of a subject found death in an advanced state of decomposition.

We developed a UPLC-MS/MS method for the quantification of the ethanol metabolites ethyl-glucuronide (EtG) and ethyl-sulfate (EtS) in blood samples. Sample preparation was a protein precipitation with acetonitrile 0.1% formic acid containing internal standards (IS) 0.5 µg mL-1 EtG-D5 and EtS-D5. Chromatographic separation was performed in a fluoro-phenyl column, mobile phases were 0.1% formic acid in water (A) and acetonitrile (B) in gradient elution from 5% B to 70% B, with 10 min run time. Ionization was performed with electrospray source at 550 °C in negative mode, quantification mass transitions of m/z EtG 221- 75, EtS 125- 80.1, EtS-D5 130- 79.9, and EtG-D5 226- 75.1. The method was linear from 0.1 to 18 µg mL-1 for EtG and 0.02 to 6 µg mL-1 for EtS, specific, with no interfering peaks during the blank samples analyses. The method was precise and accurate, the matrix effect was compensated with the IS (-12% a + 6.1%). EtG and EtS were analyzed in 28 blood samples from trauma patients with positive alcoholaemia. The concentrations of EtG and EtS ranged from 0.25 to 14.11 µg mL-1 and 0.13 to 5.53 µg mL-1, with high correlation with ethanol concentrations, r=0.739 and r=0.817 (p

Background: An increasing prevalence of alcohol consumption is a major public health problem, which has also led to an increasing number of children who have been prenatally exposed to the toxic effects of ethanol. However, obtaining reliable information on prenatal alcohol exposure through maternal self-reports has proved difficult. Aims: Our aim was to evaluate the potential for rapid screening test for measuring ethyl glucuronide (EtG), a specific alcohol metabolite, from urine samples of pregnant women. Methods: Five hundred five urine samples of pregnant women were collected anonymously from five prenatal units in two Finnish cities: a tertiary specialist antenatal clinic for pregnant women with problematic substance use (HAL), a regular hospital antenatal clinic (LCH = Lahti Central Hospital), a prenatal screening unit and two community maternity clinics (USR = user self-recruiting units). All samples were screened using rapid EtG test strips, and all positive, uncertain, and randomly selected negative samples were confirmed by quantitative analyses. The samples were also screened for cotinine and use of cannabis. Results: In this material an EtG cut-off of 300 ng/mL suggesting heavy alcohol drinking was exceeded by 7.4% (5/68) of the samples in the HAL clinic, 1.9% (4/202) in LCH, and 0.9% (2/225) in USR. A cut-off of 100 ng/mL was exceeded by 17.6% (12/68) of samples from HAL, 7.5% (16/212) from LCH, and 6.7% (15/225) from USR. Based on confirmatory quantitative analyses, there were no false negatives nor false positives in rapid EtG screening. However, 57 (11.3%) of test results were classified as uncertain. In these cases, confirmation by quantitative analyses resulted in 56.1% rate of positive values. 73% of the samples with EtG > 300 ng/mL showed positive cotinine results suggesting smoking co-occurring with alcohol intake. Conclusions: Rapid EtG tests may be an easy and inexpensive method, which may improve the possibilities for screening alcohol use among pregnant women during routine prenatal visits. Quantitative EtG analyses are recommended to confirm screening positive and uncertain cases. Trial registration: NCT04571463 Date of Registration 11/05/2020. [ABSTRACT FROM AUTHOR]

Fetal alcohol spectrum disorder (FASD) is a set of conditions resulting from prenatal alcohol exposure (PAE). FASD is estimated to affect between 2% and 5% of people in the United States and Western Europe. The exact teratogenic mechanism of alcohol on fetal development is still unclear. Ethanol (EtOH) contributes to the malfunctioning of the neurological system in children exposed in utero by decreasing glutathione peroxidase action, with an increase in the production of reactive oxygen species (ROS), which causes oxidative stress. We report a case of a mother with declared alcohol abuse and cigarette smoking during pregnancy. By analyzing the ethyl glucuronide (EtG, a metabolite of alcohol) and the nicotine/cotinine in the mother's hair and meconium, we confirmed the alcohol and smoking abuse magnitude. We also found that the mother during pregnancy was a cocaine abuser. As a result, her newborn was diagnosed with fetal alcohol syndrome (FAS). At the time of the delivery, the mother, but not the newborn, had an elevation in oxidative stress. However, the infant, a few days later, displayed marked potentiation in oxidative stress. The clinical complexity of the events involving the infant was presented and discussed, underlining also the importance that for cases of FASD, it is crucial to have more intensive hospital monitoring and controls during the initial days. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Ankara University Faculty of Medicine / Ankara Üniversitesi Tip Fakültesi Mecmuasi is the property of Galenos Yayinevi Tic. LTD. STI and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introduction:To advance the use of alcohol metabolites as biomarkers in the context of alcohol research, the present study tested the sensitivity and specificity of a commercially available urinary ethyl glucuronide (uEtG) test (DrugConfirm Advanced 80hr EtG) in a clinical research context. Methods:A community sample of heavy drinkers (N = 68) completed the 30-day Timeline Follow-Back (TLFB) interview and provided a urine sample for uEtG analysis. Analyses of sensitivity and specificity of the uEtG assay were conducted using the following outcomes: (a) past day drinking, (b) past day binge drinking (defined as ≥4 drinks for women and ≥5 drinks for men), (c) past 3-day drinking, and (d) past 3-day binge drinking. Results:The majority of participants reported past-3-day drinking (80.9%) and a sizeable minority reported past day drinking (33.8%). While uEtG-based detection of past day drinking and binge drinking was acceptable (sensitivity = 73.91%, and 83.33%; specificity = 80.00% and 66.13%, respectively), detection of any drinking and binge drinking in the past 3 days was poor (sensitivity and specificity of 43.64% and 84.62%, and 39.39% and 62.86%, respectively). Conclusions:This study contributes to the mixed findings on the validity of EtG tests, which suggest that commercial uEtG tests with conservative detection thresholds are not a reliable alcohol biomarker without corroborating self-report data. Lower detection thresholds are recommended when using uEtG as an alcohol biomarker. Efforts to reach acceptable levels of sensitivity and specificity with commercial assays hold potential to advance the measurement of alcohol intake, overcoming the pitfalls of self-report data.

Aim: Ethyl glucuronide (EtG) in hair is a direct biomarker proven to be useful for the detection of chronic excessive alcohol use. This study investigated the association of self-reported alcohol consumption with traditional biomarkers: GGT, AST, ALT, CDT and MCV in blood and a direct biomarker hair EtG in a total of 122 patients with alcohol dependence syndrome. The diagnostic accuracy of the biomarkers to differentiate heavy from non-heavy drinkers was also evaluated.Method: GGT, AST, ALT in serum was measured by Automated Chemistry Analyzer; MCV in blood was measured by Haematology Analyzer; serum CDT was analysed by ELISA and EtG in hair was evaluated by gas chromatography-mass spectrometry. The association between the biomarkers and the amount of alcohol consumed (self-reported) was determined using Spearman's rank correlation.Results: All participants showed EtG level above the cut-off (0.03 ng ⁄mg). Hair EtG showed a statistically significant linear and positive correlation with the amounts (in grams) of alcohol consumed (r = 0.60; p < 0.001). No correlation was observed among the traditional biomarkers and the quantity of alcohol consumed. Also, EtG showed an excelled ROC curve (98%) with good sensitivity (85%) and specificity (60%) to classify heavy drinkers among individuals with alcohol dependence syndrome.Conclusion: Hair EtG can be helpful to estimate retrospective alcohol consumption in long-term chronic alcohol consumption cases. Hair EtG also provides a reliable diagnostic test to detect heavy drinkers among individuals with alcohol dependence syndrome. [ABSTRACT FROM AUTHOR]

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. 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. 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. 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. 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. [Display omitted] • Alcohol intake may be a relevant contributor to the progression of fatty liver disease. • NAFLD and ALD have not been reliably distinguished by established diagnostic means. • We found relevant alcohol intake in 29% of patients with NAFLD, and 25% with MAFLD. • hEtG and uEtG showed good to excellent accuracy to detect alcohol intake. • They should be used for routine diagnostic work-up in addition to AUDIT-C. [ABSTRACT FROM AUTHOR]

Background: COVID-19 has caused nearly 1 million deaths in the United States, not to mention job losses, business and school closures, stay-at-home orders, and mask mandates. Many people have suffered increased anxiety and depression since the pandemic began. Not only have mental health symptoms become more prevalent, but alcohol consumption has also increased during this time. Helplines offer important insight into both physical and mental wellness of a population by offering immediate, anonymous, cheap, and accessible resources for health and substance use disorders (SUD) that was unobstructed by many of the mandates of the pandemic. Further, the pandemic also launched the use of wastewater surveillance, which has the potential for tracking not only population infections but also consumption of substances such as alcohol. Objective: This study assessed the feasibility of using multiple public surveillance metrics, such as helpline calls, COVID-19 cases, and alcohol metabolites in wastewater, to better understand the need for interventions or public health programs in the time of a public health emergency. Methods: Ethanol metabolites were analyzed from wastewater collected twice weekly from September 29 to December 4, 2020, in a Midwestern state. Calls made to the helpline regarding housing, health care, and mental health/SUD were correlated with ethanol metabolites analyzed from wastewater samples, as well as the number of COVID-19 cases during the sampling period. Results: Correlations were observed between COVID-19 cases and helpline calls regarding housing and health care needs. No correlation was observed between the number of COVID-19 cases and mental health/SUD calls. COVID-19 cases on Tuesdays were correlated with the alcohol metabolite ethyl glucuronide (EtG). Finally, EtG levels were negatively associated with mental health/SUD helpline calls. Conclusions: Although helpline calls provided critical services for health care and housing-related concerns early in the pandemic, evidence suggests helpline calls for mental health/SUD-related concerns were unrelated to COVID-19 metrics. Instead, COVID metrics were associated with alcohol metabolites in wastewater. Although this research was formative, with continued and expanded monitoring of population metrics, such as helpline usage, COVID-19 metrics, and wastewater, strategies can be implemented to create precision programs to address the needs of the population. [ABSTRACT FROM AUTHOR]

Abstract In this study, a liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed to detect ethyl glucuronide (EtG), which is a biomarker for monitoring alcohol consumption, and naltrexone (NTX), 6β-naltrexol (6βNTX), chlordiazepoxide (CDP), and norchlordiazepoxide (norCDP), which are analyzed to confirm the presence of medications for alcohol dependence treatment. The protein precipitation method was conducted to rapidly prepare samples. LC–MS/MS analysis was performed in the multiple-reaction monitoring mode. The analytes were separated using a Scherzo SM-C18 (2.0 × 100 mm, 3 µm) column. The calibration ranges were 5–1000 ng/mL for EtG, 6βNTX, CDP, and norCDP, and 1–100 ng/mL for NTX, with the correlation coefficients (r) being ≥ 0.994, and the weighting factor being 1/x 2. The lower limit of quantification was 1–5 ng/mL. The method was also validated for precision, accuracy, selectivity, dilution integrity, recovery, matrix effect, and stability. The developed method was successfully applied for the determination of EtG, NTX, 6βNTX, CDP, and norCDP in urine samples obtained from 49 probationers who received alcohol dependence treatment orders. The method developed herein can be used to monitor the drug-based treatment of alcohol abuse and alcohol consumption during the treatment of individuals under probation.

Alcohol biomarkers can monitor both recent and long-term drinking and provide information about drinking habits as a complement to self-reporting. Ethyl glucuronide (EtG) and phosphatidylethanol (PEth) are the most sensitive available biomarkers for this purpose. The present study aimed to collect data on both PEth and EtG in the same blood sample, in addition to ethanol, in order to evaluate the combined use of these biomarkers. Venous EDTA blood samples (n = 1149) sent to the laboratory as part of a clinical routine service for measuring PEth were investigated. PEth and EtG concentrations were analyzed using liquid chromatography–mass spectrometry methods and ethanol with an enzymatic method. Of the 1149 samples, 95 were positive for ethanol (range 0.11–3.12 g/L), 454 for EtG (1.0–9739 ng/mL), 635 for PEth (0.014–6.0 µmol/L), 534 for PEth ≥ 0.050 µmol/L, and 315 for PEth ≥ 0.30 µmol/L. EtG and PEth concentrations seemed largely independent as the coefficient of determination (r2) between PEth and EtG concentrations was 0.15. However, when the EtG concentrations were evaluated for different subgroups depending on ethanol or PEth concentrations a statistically significant difference between successive higher concentrations was observed. EtG and PEth are independent measures of recent alcohol drinking reflecting different time windows. Their combined measurement in the same blood sample is possible and will provide valuable information regarding recent alcohol consumption as a complement to self-reporting.

In the present work, we describe how MES can compromise the combined measurement and interpretation of the ethanol biomarkers ethyl glucuronide (EtG) and ethyl sulfate (EtS) [[1]]. To confirm EtG levels and to avoid the risk of misclassification of alcohol consumption based on EtG alone, a combined measurement of EtG and EtS is therefore recommended [[6]]. Keywords: alcohol marker; disinfectant; ethyl glucuronide; ethyl sulfate; mecetronium ethyl sulfate EN alcohol marker disinfectant ethyl glucuronide ethyl sulfate mecetronium ethyl sulfate e96 e98 3 04/25/23 20230515 NES 230515 To the Editor, Proper hand hygiene, including hand disinfection, is an essential tool to prevent the spreading of pathogens and diseases. [Extracted from the article]

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Ethanol, a central nervous system depressant and banned substance in horseracing, has reportedly been administered to horses prior to competition to "calm a horse's nerves." In this study, the pharmacokinetics of two metabolites of ethanol were studied to better understand the behavior of this compound in the horse and provide a scientific basis for regulation of its administration. Six horses received a single intravenous (30 mL; 1200 mg) and oral (90 mL; 3600 mg) administration of ethanol (vodka, 40% ABV) in a balanced cross-over design. Blood and urine samples were collected at various times post administration for up to 24 h. Concentrations of ethyl glucuronide and ethyl sulfate were determined using liquid chromatography-tandem mass spectrometry and pharmacokinetic analysis performed. Behavioral, locomotor activity and effects on heart rate were assessed. The maximum concentration (mean ± SD) of ethyl glucuronide was 71.5 ± 42.7 and 105.0 ± 47.5 ng/mL at 0.88 h following IV and oral administration, respectively. The maximum concentrations for the ethyl sulfate metabolite following IV and oral administration were 1.61 ± 0.60 and 3.46 ± 1.68 ng/mL, respectively. Urine concentrations of both metabolites were non-detectable by 24 h post ethyl alcohol administration. No observable behavioral responses were noted following IV or oral administration. Significant decreases in heart rate were noted at various times starting at 10 min until 4 h post administration in the oral dose group. Both ethyl glucuronide and ethyl sulfate could be useful markers for detection of illicit administration of ethanol to horses., (© 2024 The Author(s). Drug Testing and Analysis published by John Wiley & Sons Ltd.)

Abstract 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.

Fetal alcohol spectrum disorder (FASD) is a set of conditions resulting from prenatal alcohol exposure (PAE). FASD is estimated to affect between 2% and 5% of people in the United States and Western Europe. The exact teratogenic mechanism of alcohol on fetal development is still unclear. Ethanol (EtOH) contributes to the malfunctioning of the neurological system in children exposed in utero by decreasing glutathione peroxidase action, with an increase in the production of reactive oxygen species (ROS), which causes oxidative stress. We report a case of a mother with declared alcohol abuse and cigarette smoking during pregnancy. By analyzing the ethyl glucuronide (EtG, a metabolite of alcohol) and the nicotine/cotinine in the mother’s hair and meconium, we confirmed the alcohol and smoking abuse magnitude. We also found that the mother during pregnancy was a cocaine abuser. As a result, her newborn was diagnosed with fetal alcohol syndrome (FAS). At the time of the delivery, the mother, but not the newborn, had an elevation in oxidative stress. However, the infant, a few days later, displayed marked potentiation in oxidative stress. The clinical complexity of the events involving the infant was presented and discussed, underlining also the importance that for cases of FASD, it is crucial to have more intensive hospital monitoring and controls during the initial days.

Although substances incorporated by ingestion are strongly bound to hair, their loss may occur if aggressive decontamination procedures are applied, especially in highly damaged/porous hair. Evaluation of cleaning procedures using hair samples with different porosity obtained from ethanol or drug users (cocaine, heroin, methamphetamine, methadone, fentanyl, tramadol, diazepam, buprenorphine, dihydrocodeine, citalopram and trazodone). The effect of washing time and multiple wash steps with water and methanol were evaluated. Hair samples (n = 16) were selected and evaluated according to (a) the drug pattern consumption, (b) available amount, and (c) hair porosity (c1 'cosmetic treatment', c2: storage time). Six of them were soaked with an aqueous deuterated analogue solution. The samples were cut in 1‐cm segments and homogenized. All hair samples were then decontaminated one or six times with 1.5 ml of water or methanol during 1, 5, 15, 30, 60 and/or 90 min (n = 1 to 3/sample, depending on the available amount of hair). Hair extracts were then cleaned up via a solid‐phase extraction (SPE) or liquid–liquid extraction (LLE), while the washes were evaporated to dryness. All were thereafter reconstituted and analysed with routine ultra‐performance liquid chromatography–tandem mass spectrometry (UPLC‐MS/MS) methods. Although concentrations of parent drugs and/or metabolites presented a negative trend along the washing time with methanol (up to 80%), the compounds were relatively well retained in hair even after a 90 min wash (with methanol or water) in most samples, and their retention would depend mostly on the hair nature rather than their physicochemical properties (whether incorporated by ingestion and/or from external contamination). Moreover, parent drugs and/or metabolites were detected in the washes in most samples, and the ratio between hair and washes decreased along the washing time. More than 50% of the deuterated analogues soaked into hair were still present after the different washing steps. Losses were observed more frequently for long‐term stored hair samples, after decontamination with methanol for more than 30 min. Therefore, prolonged or repeated cleaning with methanol should be avoided in general procedures. [ABSTRACT FROM AUTHOR]

Abstract Aim The present study aimed to evaluate the validity of hair ethyl glucuronide concentrations compared with transdermal alcohol concentration and self‐reported alcohol use. Methods This trial included 25 adolescent and young adult females, aged 16‐24, who reported at least one heavy drinking episode (≥4 drinks) in the two weeks prior to baseline. All participants were asked to wear an alcohol biosensor over a one‐month prospective study. Detailed self‐report of drinking behaviors was assessed weekly. Estimates of blood alcohol concentration were computed from self‐report data using the National Highway and Transportation Safety Administration equation. Transdermal alcohol concentration and estimated blood alcohol concentration data were categorized into at‐risk (>0.05 g/dL alcohol) and high‐risk (>0.08 g/dL alcohol) drinking events. Hair ethyl glucuronide concentration, total number of drinking events, moderate (>0.05 g/dL) and high level (>0.08 g/dL) of transdermal alcohol concentration, and estimated blood alcohol concentration drinking events were analyzed with Spearman's rank correlation test for validity comparisons. Results No significant correlations were found between hair ethyl glucuronide values and total number, and moderate or high levels of detected drinking events by estimated blood alcohol concentration or transdermal alcohol concentration. Total number of drinking events detected and number of drinking events >0.08 g/dL using estimated blood alcohol concentration and transdermal alcohol concentration methods were significantly correlated with each other (respectively, R = .33, P

Alcohol ranks as one of the leading behavioral threats to health and life in developed countries. Alcohol abuse triggers serious social and economic effects: it contributes to higher prevalence of work-related and road accidents, as well as absence from work. The diagnosis and treatment of alcoholism still remain very difficult. Hence, the use of objective biochemical markers of alcohol abuse may contribute to earlier detection, more effective therapy and reliable teetotalism control. The aim of this study is to present the sensitive and specific biomarkers of alcohol abuse available in Poland, with particular emphasis on the practical use possibilities. Such tests may be widely used, e.g., in driving license regranting cases involving drivers whose licenses were suspended for driving when intoxicated, for the early detection of persons abusing alcohol in employment-related health controls, for abstinence monitoring during withdrawal treatment, for detecting alcohol consumption in transplant settings, for assessing the prevalence of alcohol drinking in pregnancy, as well as in autopsical examinations. The standardization of biomarkers measurement methods is essential. Moreover, concomitant disorders may pose a significant problem in the proper outcome analysis. Despite these limitations, objective biochemical markers of ethyl alcohol abuse may become helpful tools in medical care. They can play a particular role in occupational medicine diagnostics, contributing to the higher level of safety on public roads and to worker safety. Med Pr. 2021;72(2):173–84

• Polydrug use is a serious health and social problem worldwide. • A simultaneous determination in hair of DOA, ETG and synthetics opioids is described. • An analytical procedure with a single digestion, clean up and LC run is presented. • A significant reduction of times, costs and hair amount is achieved. Polydrug use is a serious health and social problem worldwide. Over the past several years, there has been an increasing tendency to combine narcotics, alcohol, sedatives, and/or stimulants. To the traditional drugs of abuse and alcohol, an increase of new abuse drugs such as synthetic opioids has been added. In the current study, the development and validation of an innovative and fast analytical procedure has been presented to determine drugs of abuse, ethyl glucuronide and synthetics opioids in 30 mg of human hair through a single digestion, purification and analysis in LC-MS/MS. A combine simple preparation of hair sample followed to a single chromatographic run of 10 min has been proposed. A full validation for 54 target analytes for the parameters of selectivity, linearity, limit of detection, limit of quantification, accuracy, precision, matrix effects, recovery, and dilution integrity was successful completed. The method was linear in different ranges with r values of at least 0.990; the value to the validated LLOQ values were in the range 0.1–100 pg/mg. The method offered satisfactory precisions (CV<15 % and accuracy ± 20 %). In conclusion, a significant reduction in the overall times of the analytical procedure and the reduction of consumables costs make this method extremely advantageous and undoubtedly useful in routine laboratory workflow analyses and open the way to the prospect of a further implementation which also includes other classes of xenobiotics. [ABSTRACT FROM AUTHOR]

This study details trends in direct alcohol biomarker concentrations from civil cases within the United Kingdom (UK). Our subject cohort in this study related to family law litigation, where an individual was subject to an alcohol monitoring order by the court. This monitoring was conducted by quantification of alcohol biomarkers Phosphatidlyethanol (PEth) in dried blood spots (DBS) and Ethyl Glucuronide (EtG) and Ethyl Palmitate (EtPa) from hair segments. In total 298 PEth cases predominantly from the South East of England during the period July 2022 to August 2023 were analysed for alcohol biomarkers in DBS and hair. Subjects alcohol intake was classified as abstinence/low alcohol consumption, moderate or excessive alcohol consumption, based on a combination of Society for Hair Testing and PEth Net guidelines. Our results indicate that 33 % of PEth concentrations were consistent with excessive alcohol use (>200 ng/mL DBS), with 36 % consistent with social or moderate alcohol use (20–200 ng/mL DBS). In relation to EtG and EtPa 23 % and 31 % of subjects were classified as excessive alcohol users respectively. This study indicates that DBS sampling of PEth is a more sensitive predictor of alcohol use, in particular, at differentiating between moderate and excessive alcohol use compared to EtG and EtPa testing in hair. The authors suggest that increased frequency in the sampling of PEth in DBS (multiple occasions per month) may provide a more accurate assessment and simplification of the interpretation criteria of alcohol patterns rather than the combined hair testing and DBS sampling that are typically requested by UK courts. • Commonly tested alcohol biomarkers were assessed in UK family law cases. • 23–33 % of subjects were classified as chronic alcohol users. • Dried blood spots provide a superior assessment of alcohol use than hair samples. [ABSTRACT FROM AUTHOR]

Ethyl glucuronide (EtG) is a minor, non-oxidative ethanol metabolite detectable in several matrices for specific periods of time. In recent years, quantification of EtG in hair has been established as the most reliable biomarker for long-term alcohol consumption, with the Society of Hair Testing offering cut-off values for assessment of both abstinence and heavy drinking. Instrumental constrains and wide inter- and intra-laboratory variability represent the ultimate barriers to widespread acceptance of hair EtG determination in the forensic context. In this study, a new analytical method for hair EtG based on gas chromatographic (GC) separation, electron impact (EI) ionization, and tandem mass spectrometry (MS/MS) detection was developed and validated. At the same time, several parameters for sample pretreatment and instrumental analysis were optimized using real hair samples obtained from different drinking subjects. A full-factorial design-of-experiment approach included procedures for hair washing, pulverization, and extraction. Rigorous multi-step washing proved not to reduce the EtG content extracted in the subsequent sample incubation. Hair pulverization with a ball mill significantly improved the EtG extraction from the keratin matrix and allowed us to reduce the time needed for the subsequent extraction step, without affecting the extraction recovery. The hair extract was derivatized with N-methyl-N-(trimethylsilyl)-trifluoroacetamide. Upon electron impact ionization of the EtG-TMS derivative, triple quadrupole mass analyzers were operated in the selected reaction monitoring (SRM) mode using the fragment m/z 405 as the precursor ion (m/z 410 for the EtG-D5 internal standard), the transitions m/z 405 → 359 and m/z 410 → 359 for quantitation, and m/z 405 → 331 and m/z 405 → 287 for qualification/confirmation, all at 10 V collision energy. The final method was fully validated and then applied to 25 real hair samples. The calibration curve proved linear between 6 and 60 pg/mg. The limit of detection (LOD) was 4 pg/mg. Intra- and inter-assay precision and accuracy tests showed a variability and bias close to 15% or lower over the entire calibration range. The new method is routinely applied in the Italian State Police’s toxicology laboratory for hair analyses addressed to exclude excessive alcohol drinking and verify the psycho-physical requirements of the personnel.

The increase in alcohol consumption in society has not only led to a number of medical issues but has also become a matter of considerable legal importance. Thus, there is both scientific interest and the necessity to diagnose alcohol abuse in the application of the provisions of the law through laboratory tests that ensure maximum objectivity. The purpose of this work is to study and compare the diagnostic performance of two of the main markers of alcohol abuse, serum carbohydrate-deficient transferrin (CDT) and Ethyl glucuronide (EtG) in a group of 336 driving under the influence (DUI) of alcohol offenders. Thus, it is possible to establish the best marker of alcohol consumption in order to assess the fitness to drive of DUI subjects.EtG was detected in 55 hair samples, while CDT was detected in 5 blood samples. Of the EtG-positive subjects 96,4% had CDT values below the cut-off. While CDT refers to an alcohol consumption of approximately the previous 10 days, EtG allows to detect an excessive alcohol consumption of the last few months. Because of these two different time-windows, EtG proves to be more reliable, since it is more difficult for subjects to change their drinking practice to test negative to toxicological analysis. The determination of Ethyl glucuronide on hair matrix is a valuable tool for the diagnosis of alcohol abuse, with high sensitivity and specificity and certainly greater reliability than traditional markers such as CDT, being a direct marker of alcohol consumption. • Chronic alcohol consumption is recognized as a major public health issue. • This study focuses on the comparison between CDT on blood and EtG on hair. • The new indicators, as EtG, are proving very useful for the diagnosis of alcoholism. [ABSTRACT FROM AUTHOR]

In this study, a liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed to detect ethyl glucuronide (EtG), which is a biomarker for monitoring alcohol consumption, and naltrexone (NTX), 6β-naltrexol (6βNTX), chlordiazepoxide (CDP), and norchlordiazepoxide (norCDP), which are analyzed to confirm the presence of medications for alcohol dependence treatment. The protein precipitation method was conducted to rapidly prepare samples. LC–MS/MS analysis was performed in the multiple-reaction monitoring mode. The analytes were separated using a Scherzo SM-C18 (2.0 × 100 mm, 3 µm) column. The calibration ranges were 5–1000 ng/mL for EtG, 6βNTX, CDP, and norCDP, and 1–100 ng/mL for NTX, with the correlation coefficients (r) being ≥ 0.994, and the weighting factor being 1/x2. The lower limit of quantification was 1–5 ng/mL. The method was also validated for precision, accuracy, selectivity, dilution integrity, recovery, matrix effect, and stability. The developed method was successfully applied for the determination of EtG, NTX, 6βNTX, CDP, and norCDP in urine samples obtained from 49 probationers who received alcohol dependence treatment orders. The method developed herein can be used to monitor the drug-based treatment of alcohol abuse and alcohol consumption during the treatment of individuals under probation. [ABSTRACT FROM AUTHOR]

As alcohol is the most common addictive substance worldwide, it is inevitable to advance the established research. New and more substantial analytical methods can be applied to reply to complex questions in legal or forensic contexts. Therefore, an analytical method for the simultaneous determination of four different alcohol biomarkers—ethyl glucuronide, ethyl sulfate, N‐acetyltaurine, and 16:0/18:1‐phosphatidylethanol—in human blood was developed, validated, and verified. Despite the different chemical properties of the analytes, a specific determination via HPLC–MS/MS was achieved using a novel type of a Phenomenex Luna® Omega Sugar column. Furthermore, all criteria for a successful validation were fulfilled according to forensic guidelines. The method proved to be linear and demonstrates selectivity and sufficient sensitivity for every biomarker. LODs obtained with this method of 2.6 ng/ml (EtG), 4.7 ng/ml (EtS), 12.5 ng/ml (NAcT), and 6.9 ng/ml (PEth) were in an acceptable range for routine applications, and the stability of all analytes over a range of 12 h is given. The verification of the new developed method was performed with authentic samples. Thus, whole blood and postmortem samples were analyzed to obtain information about the drinking behavior, which can answer complex questions regarding alcohol consumption. [ABSTRACT FROM AUTHOR]

Purpose: This review presents the current methods used for determining ethyl glucuronide (EtG) and ethyl sulfate (EtS) concentrations in postmortem specimens, including sample preparation, analysis and the role of EtG and EtS in the postmortem assessment of the extent of alcohol abuse. Methods: Papers pertaining to postmortem investigation were collected from scientific databases and reviewed. The papers were published between January 2006 and October 2020. Results: Most of the analyses involved postmortem blood and urine samples, with a few reports using other bodily specimens and tissues. The method validation was not conducted for all applications. These reports were mostly intended to present data rather than interpret them, and the lack of effort in relating these ethanol biomarkers with the cause of death and/or determination of the time of deaths due to ethanol intoxication might decrease the applicability of these makers after a promising start between 2006 and 2010. Nevertheless, by the beginning of 2020, papers investigating ethanol biomarkers were still increasing. A considerable number of methods used liquid chromatography coupled with mass spectrometry (LC-MS) techniques that require less sample preparation (e.g., protein precipitation extraction, dilution, filtration, and centrifugation). Although solid-phase extraction can be applied, only three applications were reported. Conclusions: Matrix effects can be a substantial challenge in analytical methods based on LC–MS because they directly affect the ionization of analytes. However, these problems can be avoided due to the high cutoff values used to identify positive results for these ethanol biomarkers, which are often above 0.1–1 mg/L, and using internal standards. Research on using tissue specimens is recommended as most of the reported results on this type of specimen were promising. [ABSTRACT FROM AUTHOR]

Patients with diabetes and distal symmetrical polyneuropathy (DSP) are routinely evaluated for etiologies other than diabetes, including vitamin B12 deficiency, paraproteinemia, hypothyroidism and drug or autoimmune-induced neuropathy. However, the most common cause of DSP, next to that of diabetes, is alcohol intake, which is almost never evaluated. In addition to assessment of alcohol intake based on patient history, which often leads to an underestimation of alchohol intake, markers of a high alcohol intake (elevated liver enzymes, uric acid, triglycerides, low magnesium or low folic acid levels) should be obtained. However, the test that is most likely to detect surreptitious alcohol intake is urinary ethyl glucuronide (EtG), which will detect the intake of alcohol within the previous 90 h. Detection of alcohol use is important since if alcohol consumption is not discontinued, DSP, whatever the etiology, will not improve. In addition, the use of drugs to improve symptoms of DSP (tricyclics, anti-epileptics, serotonin, norepinephrine reuptake inhibitors and analgesics) may in combination with alcohol excessively suppress respiration and cognitive function such that these drugs should not be prescribed or utilized if use of alcohol continues. In the future, all patients with DSP, and especially those with symptomatic DSP, should be biochemically screened for excessive alcohol intake and appropriate action taken. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Introduction and Aims: The increase in stress levels, social confinement, and addiction's physical consequences play an essential role in the proliferation of drug abuse. In this context, the Covid-19 pandemic produced remarkable effects on those individuals prone to addictions, especially to alcohol. Alcohol is linked to multiple dangerous conditions such as social issues, severe medical conditions, and road accidents. The determination of ethylglucuronide (EtG) in hair is frequently performed to test and monitor chronic excessive alcohol intake conditions, as it allows differentiation among low-risk/moderate drinkers, and excessive/chronic drinkers. Our study aimed to explore hair EtG levels in a controlled population to assess the impact of Covid-19 lockdown on alcohol intake along March-May 2020.Materials and Methods: EtG levels were measured in all hair samples collected in the months following April 2020 to evaluate the behaviors related to alcohol intake along with the time frame from March to May 2020. The measured concentration distributions for each month were compared with those reported in the same month during the previous 4 years (2016–2019). The dataset was built to highlight possible differences between genders, and the different categories of alcohol consumption, separately.Results: The samples collected from April to August 2020 (500 < N