Ethyl Glucuronide Concentrations In Hair Research Articles

Assessment of prenatal exposure to alcohol and gestational consumption by EtG testing in neonatal meconium and maternal hair and neonatal: First Italian national survey

Use of alcohol during pregnancy and subsequent fetal exposure can cause multiple perinatal disorders (prematurity at birth, withdrawal syndromes, tremors, hyperreflexia) and impaired physical and mental development in the later stages of life, known as fetal alcohol spectrum disorders (FADS) including as major expression of impairment fetal alcohol syndrome (FAS). Gestational drinking is usually, worldwide reported by questionnaires filled during antenatal visits, but international studies demonstrated their inconsistency for under-reporting or misreporting of toxic habits. Information concerning the true prevalence of alcohol consumption during pregnancy and consequent prenatal exposure to this teratogen is essentially lacking in Italy. To assess objective gestational consumption to alcohol and fetal exposure to it, we measured one of the most specific ethyl alcohol metabolites, ethylglucuronide (EtG), in maternal hair and neonatal meconium involving delivering mothers and newborns along the whole National Italian territory. Non-paired twenty delivering women and twenty newborns per city from hospitals of the National Health system located in the North, Centre and South Italy (including islands) were enrolled in the project. Maternal hair, 8 cm as maximum length, was collected at the end of pregnancy and analyzed as entire strand and neonatal meconium was collected within the first 24 hours after birth. Ethylglucuronide was quantified in both biological matrices through a validated gas chromatography–tandem mass spectrometry methodology. Hair EtG concentrations below 5 pg/mg hair were attributed to abstinent women, those > 5 pg/mg and below 30 pg/mg were attributed to women consuming “some” alcohol during pregnancy and ≥ 30 pg/mg were attributed to excessive drinkers. Furthermore, according to previous studies from our research group, maternal hair EtG above 11 pg/mg was shown to correspond with high probability to a neonatal meconium with EtG > 30 ng/g. Hence, it was considered as a “cut-off” matching to fetal exposure to alcohol. Up to now, 781 maternal hair and 642 meconium samples have been analyzed. Only one woman out of 781(0.1%) presented chronic excessive ethanol consumption with EtG concentration > 30 pg/mg, while the 8.2% of maternal hair presented EtG concentrations > 5 pg/mg with 1.4% > 11 pg/mg. Four newborns (0.6%) resulted prenatally exposed to ethanol with meconium EtG concentration > 30 ng/g. The highest percentage of women involved in the study (mean age + SD 34 + 5.1 years) were Italian (88%) and employed (69%). With regard to educational level, 44% had a university degree and 39% high-school education level. The four exposed newborns did not present at birth any sign of evident disability or malformation. These preliminary findings, obtained in separate cohorts of pregnant and delivering mothers and newborns showed that, currently, a negligible amount of Italian women drink during pregnancy and that a minute percentage of newborns are prenatally exposed to alcohol. The policies applied by health Italian women have acquired awareness about risks associated with drinking during pregnancy.

Ethyl glucuronide and alcohol abstinence: A correlation study in hair and fingernails to establish a cut-off value in fingernails for teetotalers

IntroductionExcessive use of alcohol increases the risk to be involved in a road traffic accident. According to many legislations, certain maximal blood-alcohol-concentrations (BAC) have to be respected to be allowed to drive on public roads. Acute alcohol intoxication is evaluated by blood analysis or analysis of the exhaled alveolar air. In many cases, evaluation of the alcohol consumption during the past months can be useful. In this light, ethyl glucuronide (EtG), a direct alcohol biomarker which can be found in keratinized matrices (hair, nails) is valuable and can be used for the long-term follow-up of alcohol consumption. Research aimTo compare the EtG concentration in hair and fingernails from teetotalers, and to propose a cut-off value for EtG in fingernails for alcohol abstinence. Material and methodsPaired samples of hair and nails were collected from participants, with a minimum age of 18 years. They all stated alcohol abstinence for at least 6 months. Participants were asked to complete a questionnaire about age, gender and the use of hair care products and nail polish. Analysis of EtG in the nail and hair samples were conducted following a validated analytical method. ResultsFrom 126 participants a hair and nail sample were collected. Of this group, 15 participants were finally not included in the study because of insufficient amount of hair or nails collected. There were more female participants (65%) and the average age of participants was 39 years. The EtG concentration in hair was below the limit of detection of 2.1 pg/mg in all but 4 samples (2.1, 2.1, 2.9, and 3.5 pg/mg). The EtG concentration in nails was below the limit of detection in 97 of the 111 samples. The concentrations in nails ranged between 2.3 and 23 pg/mg. Discussion and conclusionIn a population of 111 teetotalers, the 97.5% percentile of EtG concentrations in fingernails is 7.6 pg/mg. The highest EtG concentration observed was 23 pg/mg. These results suggest that the cut-off value for alcohol abstinence may be lower than the previous suggested 59 pg/mg and 37 pg/mg.

Validation of hair ethyl glucuronide using transdermal monitoring and self-reported alcohol use in women of childbearing potential.

AimThe present study aimed to evaluate the validity of hair ethyl glucuronide concentrations compared with transdermal alcohol concentration and self‐reported alcohol use.MethodsThis 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.ResultsNo 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 < .05; R = .42, P < .05).ConclusionOur findings indicate that, due to the number of false negatives, hair ethyl glucuronide concentrations should be used with caution for monitoring abstinence from alcohol use.

Impaired diagnostic accuracy of hair ethyl glucuronide testing in patients with renal dysfunction

The impact of renal impairment and hemodialysis on ethyl glucuronide concentrations in hair (hEtG) is not well known. Here, hEtG levels were determined by liquid chromatography-tandem mass spectrometry (LC–MS/MS) in patients presenting to the transplant outpatient clinic and compared with the self-reported alcohol consumption in an anonymous validated questionnaire. Estimated daily alcohol intake (EDI) was calculated. A total of 94 patients with varying renal function (Glomerular filtration rate (GFR) > 60 mL/min: group 1 (n = 47); GFR 30–60 ml/min: group 2 (n = 29); GFR < 30 mL/min: group 3 (n = 18)) were included in the study. Fifteen of 18 (83.3%) patients in group 3 were on dialysis. Altogether, hEtG tested positive (> 5 pg/mg) in 25.5% (n = 24) of patients, while 36.2% (n = 34) and 14.9% (n = 14) of patients reported any or regular (> 10 g/d) alcohol consumption, respectively. The median hEtG concentration of positive samples was much higher in patients in group 3 with advanced renal dysfunction or on dialysis than in patients in group 1 or 2 (group 1, 2, 3 dialysis patients: 74, 52, 145 and 155 pg/mg, respectively), although they consumed on average much less alcohol per day (median EDI group 1, 2, 3, dialysis patients: 16, 17, 3 and 3 g/d, respectively). Also, there was a significant correlation between the hEtG concentration and EDI for patients in group 1 (ρ = 0.84; p = 0.01), but not for patients in group 2 (ρ = −0.35, p = 0.39) or 3 (ρ = 0.02, p = 0.96). Furthermore, the ability of hEtG to correctly identify abstainers as such was lower for patients with advanced renal dysfunction than for the remaining patients (specificity for group 1, 2, 3: 92%, 87%, 82%, respectively). So, monitoring hEtG concentration was less reliable in patients with advanced renal dysfunction or on hemodialysis and by far overestimated the amount of alcohol consumed.

Improved measurement of ethyl glucuronide concentrations in hair using UPLCMS/MS for the evaluation of chronic ethanol consumption

The presence of Ethyl glucuronide (EtG) in hair provides a strong indication of ethanol consumption and its investigation is of interest in both clinical and forensic contexts because of the wide window of detection. However, due to the possibility of false negative results in cases of small ethanol intake or excessive hair washing, the combined measurement of ethyl palmitate (EtP) with EtG could be useful. In this study, a sensitive UHPLC-MS/MS procedure for the measurement of EtG in hair was developed and validated, using optimized sample preparation and chromatographic separation. Milled hair was extracted with water for 24 h at room temperature, followed by clean-up of the extract by ion-exchange solid phase extraction (SPE). Extraction was highly efficient, with yield of 96.93–101.06%. Chromatographic separation was performed with a Fluoro-Phenyl stationary phase. The assay was linear from 4 to 500pgmg−1, with accuracy in the range of 100.30–106.16%. Matrix effects (−0.87 to 5.89%) were adequately compensated by the use of deuterated EtG as internal standard. EtG was measured in hair samples of 46 volunteers, and results were compared with hair concentrations of ethyl palmitate (EtP) and the score in the AUDITC questionnaire. EtG hair concentrations were significantly correlated to the AUDIT-C classification (rs=0.365, p<0.05), but not to EtP hair levels. The diagnostic performance of EtG hair concentrations to identify excessive or moderate ethanol use was similar to the capability of AUDIT-C to identify severe and high health risk (Kappa, p=0.013). The developed assay is suitable for clinical use, providing a useful tool to evaluate chronic ethanol consumption.

Ethyl Glucuronide Concentration in Hair of Detainees: A Preliminary Study.

Through the measurement of ethyl glucuronide in hair (hETG), it is possible to assess chronic alcohol abuse over time. In this paper, we present a study on hETG in Italian prison inmates. Analyses were performed by LC-MS according to a previously published method. Results were evaluated using the cut-offs established by the Society of Hair Testing. Positives samples (ETG>30pg/mg) accounted for 6% of all subjects, with concentrations ranging from 42pg/mg up to 270pg/mg, abstinent subjects (ETG<7pg/mg) accounted for 88%, and moderate alcohol consumption (7<ETG<30pg/mg) for 6% of the subjects. No females displayed ETG values above 30pg/mg. Among positive samples, only two subjects did not declare heavy alcohol consumption and werefound strongly positive at 210 and 270pg/mg. To the best of our knowledge, this represents the first study on ETG hair concentration on prison inmates.

Chronic alcohol abuse may lead to high skin iron content, but not to hepatic siderosis

IntroductionIn forensic cases, ante mortem chronic alcohol abuse can be of central importance in clarifying circumstances of death. However, reliable markers of alcohol consumption, which are still available postmortem, are needed. In addition to medical history data which may not be always authentic, the determination of ethyl glucuronide (EtG) in hair as a promising parameter is of no value in cases of missing or cosmetically treated hair. On the other hand, there exist reports that iron ions accumulate in liver tissue (siderosis) during chronic, excessive alcohol consumption, which, therefore, may be useful to serve as alcohol abuse correlate. However, the influence of ethanol on iron stored in the liver has not been adequately investigated and the study situation appears to be inconsistent. AimsThe aim of the present study was to assess the suitability of assaying iron concentrations in liver and other tissues as postmortem alcoholism marker. MethodsThe iron concentration in tissue samples (liver, brain, skin, pancreas, spleen), vitreous fluid and blood taken during autopsy was analyzed by atomic absorption spectroscopy. The analytical method has been validated before. Cases were divided into two groups: chronic alcohol abusers and non-chronic alcohol consumers including total abstainers using ethyl glucuronide levels in hair as well as anamnestic data as criteria. ResultsNo elevated iron concentrations in the liver of chronic alcohol abusers were detected. Surprisingly, the iron concentration in skin tissue was found to be significantly higher in cases of chronic alcohol abuse, independent on whether fatty liver or liver cirrhosis was present (as diagnosedduring autopsy).In 18.5% of the cases, chronic alcohol abuse was not confirmed by the EtG concentration in hair. Thus, anamnestic data should not be overestimated. ConclusionThe general assumption that chronic alcohol abuse induces hepatic siderosis, i.e. high iron concentrations in liver tissue, has not been supported by results of the present study. However, there seems to exist a correlation between chronic alcohol abuse and high iron concentrations in the skin.

How to Interpret Hair EtG Concentrations in Individuals with High Body Mass Index? Brief Commentary on: Influence of Body Mass Index on Hair Ethyl Glucuronide Concentrations.

Body mass index (BMI) is a variable that complicates the interpretation of the alcohol metabolite ethyl glucuronide (EtG) in hair. However, direction on how EtG should currently be interpreted within individuals consuming moderate and excessive daily amounts of alcohol related to their BMI is lacking. In light of interpretation of EtG in individuals with high BMI, we present post hoc analysis of earlier data regarding the effect of BMI on hair EtG concentrations.

Variability on ethyl glucuronide concentrations in hair depending on sample pretreatment, using a new developed GC–MS/MS method

Quantitative determination of ethyl glucuronide in keratin matrix, particularly in hair samples, provides a significant contribution to the evaluation of the extent of ethanol intake. The first-choice method to carry out this analysis is LC–MS/MS, but other techniques may be used. The aim of this work is: a) to develop and validate a GC–MS/MS method for ethyl glucuronide determination in hair; b) to compare GC–MS/MS and LC–MS/MS in analysis of real samples; c) to compare EtG concentration obtained after hair cutting and pulverization.About 30 mg hair samples were washed, pulverized and soaked in 1 ml deionized water. After incubation, the solution was purified through a SPE anion exchange cartridge; the eluate was dried under nitrogen stream, derivatized with PFPA and reconstituted in n-hexane. Then, the sample was injected in the GC–MS/MS system, operating in negative chemical ionization mode and in selected reaction monitoring. The two most intense transitions were used to monitor ethyl glucuronide and deuterated internal standard. All the validation parameters fulfilled the international acceptance criteria. LOD and LOQ were set at 2.0 and 3.0 pg/mg respectively.This method was applied to 194 hair samples collected from teetotallers and alcohol consumers and represents a suitable alternative to LC–MS/MS for the determination of EtG in hair samples, in particular when scarce quantity of hair is available. This study confirmed that pulverization of hair increases the concentration of EtG, but some variability of EtG levels remains probably due to the presence of non-homogeneous material even though pulverization.

Assessment of ethyl sulphate in hair as a marker for alcohol consumption using liquid chromatography-tandem mass spectrometry.

Ethyl glucuronide (EtG) and ethyl sulphate (EtS) are 2 non-oxidative and direct metabolites of ethanol. EtG is known to accumulate in hair and has proved to be a reliable biomarker for detection of chronic alcohol consumption. EtS has been analysed in blood and urine but has never been reported in hair. This article presents the first analytical assay based on liquid chromatography coupled to tandem mass spectrometry for the quantification of EtS in hair. Sample preparation, chromatographic, and mass spectrometric parameters, such as solid-phase extraction, column type, and transitions were optimised. The method was validated according to the guidelines of the European Medicine Agency, fulfilling the requirements for limit of quantification (LOQ), linearity, accuracy, precision, carry-over, matrix effects, and recovery. Linearity ranged from 5 to 500pgmg-1 and the LOQ was achieved at 5pgmg-1 . The novel method was successfully applied to hair samples (n=40) from patients treated for alcohol use disorders. EtS concentrations in hair ranged from 24 to 1776pgmg-1 , while EtG concentrations in hair ranged from 1 to 1149pgmg-1 . Hair concentrations of EtS and EtG were compared to assess the relationship between both biomarkers. There was a significant and positive correlation between EtS and EtG in hair, suggesting that EtS can be used as a biomarker for alcohol consumption. Relatively high basal EtS levels were observed in alcohol-abstinent persons, comparable to what has been reported for EtG. The developed analytical procedure offers an alternative method to prove alcohol consumption using hair analysis.

The determination of ethyl glucuronide in hair: Experiences from nine consecutive interlaboratory comparison rounds

The increasing request for hair ethyl glucuronide (HEtG) in alcohol consumption monitoring according to cut-off levels set by the Society of Hair Testing (SoHT) has triggered a proficiency testing program based on interlaboratory comparisons (ILC). Here, the outcome of nine consecutive ILC rounds organised by the SoHT on the determination of HEtG between 2011 and 2017 is summarised regarding interlaboratory reproducibility and the influence of procedural variants. Test samples prepared from cut hair (1mm) with authentic (in-vivo incorporated) and soaked (in-vitro incorporated) HEtG concentrations up to 80pg/mg were provided for 27–35 participating laboratories. Laboratory results were evaluated according to ISO 5725-5 and provided robust averages and relative reproducibility standard deviations typically between 20 and 35% in reasonable accordance with the prediction of the Horwitz model. Evaluation of results regarding the analytical techniques revealed no significant differences between gas and liquid chromatographic methods In contrast, a detailed evaluation of different sample preparations revealed significantly higher average values in case when pulverised hair is tested compared to cut hair. This observation was reinforced over the different ILC rounds and can be attributed to the increased acceptance and routine of hair pulverisation among laboratories. Further, the reproducibility standard deviations among laboratories performing pulverisation were on average in very good agreement with the prediction of the Horwitz model. Use of sonication showed no effect on the HEtG extraction yield.

Usefulness of hair analysis and psychological tests for identification of alcohol and drugs of abuse consumption in driving license regranting

The implementation of the points-based driving license helps to change the drivers’ behavior and is related to a reduction of traffic accidents and fatalities. In Spain, when a driver loses all points, the driving license is revoked, so the driver must enroll on a Driver Awareness and Re-education (DARE) course. However, at the moment offenders are not submitted to any test to confirm absence of alcohol or drugs of abuse consumption, even when 9% of Spanish drivers lose their driving license for driving under the influence (DUI). The objective of this pilot study was the comparison of the usefulness of psychological tests and hair analysis to identify those individuals with a chronic consumption of alcohol and drugs of abuse among drivers performing DARE courses. Volunteers were submitted to the AUDIT and DAST-10 tests. Also a hair sample was collected and analyzed for ethylglucuronide (EtG) (LOQ 5pg/mg) and 35 licit and illicit drugs (LOQ 5–50pg/mg) by LC–MS/MS. Sixty-one participants with a mean age of 37.2±11.6years, and mainly men (90.2%), were recruited and performed AUDIT and DAST-10 tests. All hair samples were analyzed for EtG and 17 samples for licit and illicit drugs. Mean AUDIT score was 9.6 (SD=7.5), showing a value ≥8 (indicator of hazardous and harmful alcohol use) in 52.4% of cases. Mean DAST-10 score was 2.9 (SD=3.3), but a score ≥6 was detected in 21.3% of cases (indicating drug abuse or dependence). Twenty-two samples were positive for EtG, 8 for drugs of abuse (8 cocaine, 2 opioids, 1 amphetamines, 1 cannabis), and 3 for medicines. EtG concentration (20.7–1254.1pg/mg) was higher than the Society of Hair Testing (SoHT) cut-off for chronic alcohol consumption (≥30pg/mg) in 21 cases. All positive cases for methadone and cannabis, and half of positive cases for opioids and cocaine presented higher concentrations than SoHT cut-offs for chronic consumption. Higher AUDIT score and higher EtG concentration in hair were statistically associated with declaration of alcohol consumption ≥4 times/month and with previous fine for DUI of alcohol. In addition, AUDIT scores and EtG concentration in hair had a moderate but significant Spearman correlation (r=0.331, p<0.05). The combination of psychological tests and hair analysis seems to be a promising tool to identify individuals with chronic and problematic consumption of alcohol and drugs of abuse. Moreover, their application during driving license regranting procedures could increase the effectiveness of DARE courses, reduce recidivism and improve road safety.

Ethyl glucuronide in keratinous matrices as biomarker of alcohol use: A correlation study between hair and nails

To quantify alcohol use, objective, specific and sensitive long-term alcohol markers are necessary. Ethyl glucuronide (EtG), a direct metabolite of alcohol, accumulates in keratinous matrices such as hair and nails, and is a specific and sensitive long-term biomarker for the detection of chronic alcohol consumption. So far, research has primarily focused on the detection of EtG in hair, and studies on its measurement in nails are scarce. In this article, we assessed EtG concentrations in hair, finger- and toenails from the same individuals in order to evaluate the direct correlation between the matrices. To this end, a total amount of 45 hair, 41 fingernail, and 13 toenail samples were collected from patients treated for alcohol use disorders at two psychiatric centers in Belgium. Samples were analyzed by gas chromatography-tandem mass spectrometry. Hair EtG concentrations ranged from <LLOQ to 1149pg/mg (median=164pg/mg, IQR [42; 283]). Fingernail EtG concentrations ranged from <LLOQ to 4090pg/mg (median=250pg/mg, IQR [74; 645]). Toenail EtG concentrations ranged from 127 to 3792pg/mg (median=687pg/mg, IQR [379; 1370]). EtG levels in hair and nails were significantly and positively correlated (p-values<0.001, r=0.70 and r=0.62, respectively). Higher concentrations were present in finger- and toenails compared to hair, which might be attributed to the slower growth rate of nails, resulting in increased accumulation of EtG. Hence, nail analysis may be interesting when low concentrations of EtG are expected, e.g. to discriminate between teetotalers and social drinkers. In addition, the current study proposes preliminary cut-off values for EtG concentrations in fingernails: >123pg/mg for chronic excessive alcohol consumption, 59–123pg/mg for moderate alcohol consumption, and <59pg/mg for alcohol abstinence. In light of these results, nails may be a useful alternative to hair samples for monitoring of long-term alcohol consumption, e.g., in cases where hair is not available. Further studies are needed to establish cut-off values for EtG levels in nails.

Influence of Body Mass Index on Hair Ethyl Glucuronide Concentrations.

Analysis of ethyl glucuronide(EtG) concentrations in hair is increasingly used to estimate the consumption of alcohol of the prior months. Linear correlations between the amount of alcohol consumed and the concentration of EtG in hair have been reported, and several variables that may influence this correlation have been investigated: e.g. cosmetic hair treatments, gender influences or hair color. Here, we investigate the influence of body mass index (BMI) on this correlation. A post hoc analysis on the influence ofBMI on the relation between amounts of alcohol consumed and the measured EtG concentrations in hair in 199 participants. Our data show higher EtG concentrations in participants with high BMI (≥25) compared to participants with low BMI (<25) (P =0.001) across a wide range of amounts of alcohol consumed. We conclude that BMI should be taken into account when interpreting hair EtG concentrations. Ethyl glucuronide concentrations in hair (hEtG) can be used to estimate the consumption of alcohol of the prior months. Body mass index (BMI) influences this relation and BMI should be taken into account when interpreting hEtG concentrations in participants with high BMI (≥25) compared to participants with low BMI (<25).

Ethyl glucuronide concentrations in hair: a controlled alcohol-dosing study in healthy volunteers.

Ethyl glucuronide (EtG) is a minor phase II metabolite of alcohol that accumulates in hair. It has been established as a sensitive marker to assess the retrospective consumption of alcohol over recent months using a cut-off of ≥7 pg/mg hair to assess repeated alcohol consumption. The primary aim was to assess whether amounts of alcohol consumed correlated with EtG concentrations in hair. Additionally, we investigated whether the current applied cut-off value of 7 pg/mg hair was adequate to assess the regular consumption of low-to-moderate amounts of alcohol. A prospective controlled alcohol-dosing study in 30 healthy individuals matched on age and gender. Individuals were instructed to drink no alcohol (N = 10), 100 g alcohol per week (N = 10) or 150 g alcohol per week (N = 10) for 12 consecutive weeks, before and after which hair was collected. Throughout the study, compliance to daily alcohol consumption was assessed by analyzing urine EtG three times weekly. Participants in the non-drinking group had median EtG concentrations of 0.5 pg/mg hair (interquartile range (IQR) 1.7 pg/mg; range < 0.21-4.5 pg/mg). Participants consuming 100 and 150 g alcohol per week showed median EtG concentrations of 5.6 pg/mg hair (IQR 4.7 pg/mg; range 2.0-9.8 pg/mg) and 11.3 pg/mg hair (IQR 5.0 pg/mg; range 7.7-38.9 pg/mg), respectively. Hair EtG concentrations between the three study groups differed significantly from one another (p < 0.001). Hair EtG concentrations can be used to differentiate between repeated (low-to-moderate) amounts of alcohol consumed over a long time period. For the assessment of repeated alcohol use, we propose that the current cut-off of 7 pg/mg could be re-evaluated.

Influence of repeated permanent coloring and bleaching on ethyl glucuronide concentrations in hair from alcohol-dependent patients

BackgroundEthyl glucuronide (EtG), a minor metabolite of alcohol, is used as a sensitive marker in hair to detect the retrospective consumption of alcohol. The proximal 0–3cm hair segment is often used for analysis, providing information on alcohol consumption over the past 3 months. Using more distal segments would allow the detection of alcohol consumption over longer time periods, thereby addressing the chronicity of the consumption. In view of this, permanent coloring and bleaching were shown in vitro to alter EtG concentrations in hair, but no in vivo studies are available to prove or disprove this. AimsTo investigate the influence of repeated bleaching and permanent coloring on EtG concentrations in vivo and to assess the stability of EtG concentrations in distal compared to proximal hair segments. MethodsHair samples from alcohol-dependent patients with uncolored/unbleached (N=4), permanent coloration (N=5) and bleached hair (N=5) were analyzed in two to six 3cm long segments for EtG concentrations, and alcohol consumption and hair cosmetic treatments were assessed. ResultsWe observed that hair bleaching and permanent coloring reduces EtG concentrations by 82±11% and 65±24%, respectively, with correlations between the number of cosmetic treatments and the decrease in EtG concentrations. EtG remained stable in untreated hair samples up to 18cm. ConclusionsEtG is a sensitive marker to assess chronic alcohol consumption up to 18 months in alcohol-dependent patients with no cosmetic hair treatments. However, in alcohol-dependent patients who color or bleach their hair, care should be taken when interpreting EtG measurements.

The influence of cleansing shampoos on ethyl glucuronide concentration in hair analyzed with an optimized and validated LC–MS/MS method

Ethyl glucuronide (EtG) is widely used as a marker for assessment of alcohol consumption behavior. In this study the influence of special cleansing shampoos on ethyl glucuronide concentrations in hair was investigated. For that purpose an optimized LC–MS/MS method was developed using a Hypercarb™ porous graphitic carbon (PGC) column and validated according to the guidelines of the German Society of Toxicological and Forensic Chemistry (GTFCh). Twenty-five hair samples of persons with known alcohol consumption behavior were investigated (21 positive samples and 4 blank samples). The hair samples were divided into two strands of hair and were analyzed after treatment with one out of four cleansing shampoos and without shampoo treatment. EtG concentrations in hair did not show any significant differences after a single application of the different cleansing shampoos. EtG was still detectable in all the positive hair samples without significant concentration change. These results clearly demonstrated that a single application of the tested cleansing shampoos did not remove EtG from hair and therefore had no influence on EtG concentration in analytical hair analysis.

P8: The influence of cleansing shampoos on ethyl glucuronide concentrations in hair analyzed with an optimized and validated LC-MS/MS method

Introduction Ethyl glucuronide (EtG) has been established as a routine biomarker for monitoring alcohol abuse or abstinence. Recently, social networks and blogs have become a platform for sharing information on how to avoid EtG detection through bleaching, dyeing, shaving or straightening the hair. In this study, we investigated the influence of four special cleansing shampoos that promise to ‘detox’ and thereby reduce ethyl glucuronide concentrations in hair. Methods An optimized LC-MS/MS method was developed using a hypercarb ® porous graphitic carbon (PGC) column and validated according to the guidelines of the German Society of Toxicological and Forensic Chemistry. For the comparative analysis of the present study, 25 real case hair samples were analyzed in total with 21 positive hair samples covering a concentration range of EtG ( c ETG ) from 7-288 pg/mg and 4 negative hair samples with c ETG Results EtG concentrations in hair did not show any significant differences after a single application of the various cleansing shampoos. EtG was still detectable in all hair samples at equal concentration levels, respectively. These results clearly demonstrated that a single application of the tested cleansing shampoos did not remove or reduce EtG from hair and therefore had no influence on EtG concentrations. Conclusion The reported LC-MS/MS method for the analysis of EtG in hair has been used to study the influence of so-called “detox” shampoos on EtG concentration in hair. This study showed that none of the four selected shampoos influenced the detected concentration of EtG in hair. These results demonstrate that these shampoos do not remove EtG from hair after a single application.

O14: Alcohol consumption vs. hair EtG concentration in alcohol dependent individuals: Role of gender differences

Introduction Ethyl glucuronide (EtG), a minor alcohol metabolite, accumulates in hair and is proposed as a stable marker for the detection of excessive alcohol consumption above a cut-off level of 30 pg/mg hair. A correlation between drinking behavior and EtG hair concentrations is observed, but variability exists, which could partially be explained by gender differences. Aims To investigate (1) the correlation between alcohol consumption and hair EtG in alcohol dependent patients, and (2) the effect of gender differences on this correlation. Methods The first 0–3 cm hair segments were analyzed using gas chromatography mass spectrometry (GC-MS) in negative ion chemical ionization (NICI) mode. Detailed retrospective alcohol consumption was obtained on a day per day basis over the last 3 months using the Timeline Follow Back Interview in alcohol dependent individuals starting an alcohol detoxification program. Results Thirty-six Caucasian alcohol dependent individuals (25 males/11 females; mean age 45 ± 8 years; mean BMI 25 ± 6) participated. Median alcohol consumption over the last three months (90 days) was 13050 g alcohol (range 65 – 650 g alcohol/ day). Hair EtG concentrations varied between 32 pg/mg hair and 662 pg/mg hair. There was a good linear and positive correlation between hair EtG and the amounts of alcohol consumed (Pearson r = 0.83; p r = 0.82; p r = 0.78; p = 0.005). Conclusions Our data confirm the good linear correlation between hair EtG and amounts of alcohol consumed, extending findings to alcohol dependent individuals. Additionally, we show that gender differences do not play a crucial role in the interpretation of hair EtG values for evaluating the amounts of alcohol consumed. As a result, no separate cut-off values are necessary for male and female alcohol abusers. These findings should be replicated in recreational alcohol consumers to assess possible gender differences in the interpretation of lower hair EtG values.

Improved liquid chromatography-tandem mass spectrometric method for the determination of ethyl glucuronide concentrations in hair: Applications to forensic cases

Ethyl glucuronide (EtG) is a direct marker of ethanol consumption, and its assay in hair is an efficient tool for chronic alcoholism diagnosis. In 2012, the Society of Hair Testing proposed a new consensus for hair concentrations interpretation, strongly advising the use of analytical methods providing a limit of quantification of less than 3pg/mg. The present work describes the optimization and validation of a previously developed liquid chromatography-tandem mass spectrometric method in order to comply with this recommendation. The concentration range of this improved method is from 3 to 1,000pg/mg. Some cases are then described to illustrate the usefulness of hair EtG: a forensic post-mortem case and two cases of suspension of driving licences. Finally, hair samples of some teetotallers (n = 10) have been analyzed, which allowed neither to quantitate nor to detect any trace of EtG.