Rapid urine screening for ethyl glucuronide from pregnant women as a tool for detecting prenatal alcohol exposure.

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.

Clinical usefulness of a urine dipstick to detect ethyl glucuronide (EtG): A quantitative clinical study in healthy young female volunteers.

The metabolite of ethanol, ethyl glucuronide (EtG), reflects alcohol intake longer than ethanol and is used as a biomarker in clinical settings to detect alcohol use. We aimed to assess the clinical usefulness in a low-to-moderate alcohol intake setting and validate a new urine EtG dipstick. A three-way, open, cross-over trial was conducted. Data were collected from January to June 2019. Among 12 healthy female volunteers, we quantified urine EtG and used a dipstick following intake of either one, two or four units of alcohol. Main outcomes were concentrations of EtG in urine and serum, and creatinine and ethanol in serum. EtG in urine was determined dichotomously by dipsticks at two different thresholds and by mass spectrometry used as gold standard. EtG in urine was quantifiable up to 24 hours after alcohol intake. In some individual cases, EtG was quantifiable up to 72 hours at low concentrations. The dipstick detected EtG in urine up to 24 hours. At thresholds of 1000 and 1500 ng/mL, the dipsticks had a specificity of 100% (both), while sensitivity was 84% and 69%, respectively. The sensitivity of the dipsticks was insufficient to support a screening purpose in this setting of low-to-moderate alcohol intake.

Rapid Screening of Urinary 1-Hydroxypyrene Glucuronide by Multisegment Injection-Capillary Electrophoresis-Tandem Mass Spectrometry: A High-Throughput Method for Biomonitoring of Recent Smoke Exposures.

Urinary 1-hydroxypyrene (HP) is a widely used biomarker of polycyclic aromatic hydrocarbon exposure relevant for biomonitoring the deleterious health impacts from tobacco smoke and ambient air pollution, as well as the hazards of certain occupations. Conventional methods for urinary HP analysis based on liquid chromatography with native fluorescence detection or tandem mass spectrometry (MS/MS) and gas chromatography-mass spectrometry (GC-MS) are limited by low sample throughput and complicated sample workup protocols that are prone to bias. Herein, we introduce a high throughput method to directly analyze the intact glucuronide conjugate of HP (HP-G) in human urine after a simple acidified ether extraction procedure when using multisegment injection-capillary electrophoresis-tandem mass spectrometry (MSI-CE-MS/MS). Multiplexed analyses of 13 independent urine extracts are achieved in a single run (<3 min/sample) with stringent quality control while avoiding enzyme deconjugation and precolumn chemical derivatization. Method validation demonstrates good technical precision (CV = 7.7%, n = 45) and accuracy with a mean recovery of (93 ± 3%) for urinary HP-G at three concentration levels with adequate detection limits (7 ng/L, S/N = 3). An interlaboratory method comparison of urine samples collected from firefighters deployed in the 2016 Fort McMurray wildfire also confirms good mutual agreement with an acceptable negative bias (mean bias = 15%, n = 55) when measuring urinary HP-G by MSI-CE-MS/MS as compared to total hydrolyzed urinary HP by GC-MS due to the low residual levels of free HP and its sulfate conjugate. This multiplexed separation platform is optimal for large-scale biomonitoring studies of air pollution relevant to global health as well as occupational smoke exposures in firefighters susceptible to dermal PAH absorption when using personal protective equipment.

ß-Glucuronidase- and OATP2B1-mediated drug interaction of scutellarin in Dengzhan Xixin Injection: A formulation aspect.

Scutellarin is the major and active constituent of Dengzhan Xixin Injection (DZXX), a traditional Chinese medicine prepared from the aqueous extract of Erigeron breviscapus and widely used for the treatment of various cerebrovascular diseases in clinic. In present study, the possible pharmacokinetic differences of scutellarin after intravenous administration of scutellarin alone or DZXX were explored. Additional, the potential roles of ß-glucuronidase (GLU) and OATP2B1 in drug-drug interaction (DDI) between scutellarin and constituents of DZXX were further evaluated in vitro. The plasma concentration, urinary and biliary excretion of scutellarin in rats after administration of DZXX, were significantly higher than those received scutellarin, while pharmacokinetic profile of Apigenin 7-O-glucuronide (AG) in rats was similar no matter AG or DZXX group. Furthermore, higher concentration in brain and plasma, however, lower level of scutellarin in intestine were observed after intravenous administration of DZXX. Finally, AG and caffeoylquinic acid esters were found to significantly inhibit GLU and OATP2B1 in vitro, which might explain, at least in part, the pharmacokinetic DDI between scutellarin and other chemical constituents in DZXX. The findings provided deep insight into the prescription-formulating principle in DZXX for treating the cerebrovascular diseases.

Persistent Urinary Ethyl Sulfate in the Absence of Urinary Ethyl Glucuronide in a Patient with Alcohol Use Disorder Who Claimed Abstinence.

A 48-year-old nurse with an alcohol use disorder history was being monitored in a professional health program. She consistently produced low-to-moderate urinary ethyl sulfate (EtS) concentrations in the absence of detectable urinary ethyl glucuronide (EtG), blood phosphatidylethanol and breath alcohol. She denied intentional ethanol consumption. After prolonged monitoring in a drug treatment program, including a period in a controlled environment, we concluded that this individual's urinary EtS likely resulted from anatomical and microbial factors related to Roux-en-Y gastric bypass surgery, with possible contributions from hidden dietary sources of ethanol. We have no definitive explanation for the lack of urinary EtG.

Urinary ethyl glucuronide (uEtG) as a marker for alcohol consumption in liver transplant candidates: a real-world cohort.

BACKGROUND: In order to reduce alcohol relapse after liver transplantation (LT), the German national guidelines for waiting-list maintenance and organ allocation demand a minimum 6-month period of alcohol abstinence pre-LT, confirmed by measuring urinary ethyl glucuronide (uEtG). METHODS: Between January 2015 and June 2016, uEtG was measured at least once in 339 cirrhotic patients with an indication for LT at the University Medical Center Mainz. uEtG was measured with an enzyme-linked immunosorbent assay (ELISA) screening test (cutoff value: 500 µg/L). For uEtG values ≥ 500 µg/L, liquid chromatography-mass spectrometry (LC-MS/MS) was performed as a confirmatory assay. Data were collected prospectively in a transplant database. RESULTS: Of the 339 potential liver transplant candidates, uEtG was negative in 86.4 %. Most patients were male (64.3 %), with an average age of 56.42 ±â€Š10.1 years. In the multivariate analysis, mean corpuscular volume (p = 0.001), urinary creatinine (p = 0.001), gamma-glutamyl transferase (p = 0.001), and hemoglobin (p = 0.003) were significantly associated with a positive uEtG test result. The sensitivity of the ELISA screening test was 100 % for uEtG values > 2000 µg/L, as confirmed by LC-MS/MS. CONCLUSION: uEtG is an effective parameter to reveal alcohol consumption by patients on the waiting list for LT. The sensitivity of the ELISA is excellent for uEtG values > 2000 µg/L, for which LC-MS/MS confirmation could be omitted.

Fetus morphology changes by second-trimester ultrasound in pregnant women drinking alcohol.

Fetal alcohol spectrum disorders (FASDs) are a group of negative conditions occurring in children exposed to alcohol during gestation. The early discovery of FASD is crucial for mother and infant follow-ups. In this study, we investigated in pregnant women the association between urine ethylglucuronide (EtG-a biomarker of alcohol drinking) and indicators of the physical characteristics of FASD by prenatal ultrasound in the second trimester of gestation. We also correlated these data with the AUDIT-C, T-ACE/TACER-3, TWEAK, and food habit diary, screening questionnaires used to disclose alcohol drinking during pregnancy. Forty-four pregnant women were randomly enrolled and examined for ultrasound investigation during the second trimester of gestation. Urine samples were provided by pregnant women immediately after the routine interviews. EtG determinations were performed with a cutoff established at 100 ng/mL, a value indicating occasional alcohol drinking. Fifteen of the enrolled pregnant women overcame the EtG cutoff (34.09%). Analysis of variance (ANOVA) revealed that the fetuses of the positive EtG pregnant women had significantly longer interorbital distance and also significantly increased frontothalamic distance (P's < 0.02). Quite interestingly, no direct correlation was found between EtG data and both food diary and AUDIT-C. However, a significant correlation was observed between urinary EtG and T-ACE (r = 0.375; P = 0.012) and between urinary EtG and TWEAK (r = 0.512; P < 0.001) and a concordance with all questionnaire for EtG values higher than 500 ng/mL. This study provides clinical evidence that the diagnosis of maternal alcohol consumption during pregnancy by urine EtG may disclose FASD-related damage in the fetus.

Lessons learned from a case of tert-butyl glucuronide excretion in urine - "New" psychoactive alcohols knocking on the back door?

BACKGROUND: Ethyl glucuronide (EtG) in urine is considered a marker of recent ethanol consumption or ethanol exposition. tert-Butanol is primarily used as a solvent and intermediate chemical. Like tert-amyl alcohol, tert-butanol is discussed in Internet forums as ethanol replacement. We discuss false-positive immunological EtG screenings by excretion of different alcohol glucuronides (EtG homologs), mainly tert-butyl glucuronide in urine of a polytoxikomanic in-patient. METHODS: Three consecutive urine samples from an in-patient with a long history of multiple substance abuse including solvents were analyzed by DRI EtG enzyme immunoassay (ThermoFisher Scientific Microgenics) on a Beckman Coulter AU680 analyzer, an in-house LC-MS/MS for EtG, 1-propyl, 2-propyl, 1-butyl, 2-butyl, and tert-butyl glucuronide, and an in-house headspace GC-FID of free congener substances methanol, 1-propanol, 2-butanone, 2-butanol, isobutanol, 1-butanol, 3-methyl-1-butanol, 2-methyl-1-butanol, and additionally for ethanol, acetone, 2-propanol, tert-butanol and 2-methyl-2-butanol. RESULTS: EtG immunoassay yielded two positive urine samples (0.2 and 0.6mg/L or 0.1 and 0.2mg/g creatinine; cut-off 0.1mg/L) which were tested EtG negative by LC-MS/MS (cut-off 0.1mg/L) but positive for tert-butyl glucuronide (3.7 and 27.1mg/L), 2-butyl glucuronide (1.1 and 3.5mg/L), and 2-propyl glucuronide (0.1 and 0.4mg/L). Headspace GC-FID detected tert-butanol (0.97 and 4.01mg/L), methanol (0.96 and 0.62mg/L), 2-butanone (0.84 and 1.65mg/L), and 2-butanol (0.04 and 0.09mg/L), but no ethanol and no 2-methyl-2-butanol. CONCLUSION: Cross-reaction of EtG homologs, mainly tert-butyl glucuronide after suspected tert-butanol or isobutane abuse, explains the false-positive EtG immunoassay findings. Future investigations could address the usefulness of alcohol glucuronides (EtG homologs) in urine as (a) biomarkers of an exposition to alkans or their corresponding alcohol metabolites and (b) as markers for using "old"-well known alcohols like tert-butanol or tert-amyl alcohol as easy to obtain, cheap, potent and "undetectable" ethanol replacements or "New" Psychoactive Alcohols.

Introduction of sample tubes with sodium azide as a preservative for ethyl glucuronide in urine.

Ethyl glucuronide (EtG) is a direct alcohol marker, which is widely used for clinical and forensic applications, mainly for abstinence control. However, the instability of EtG in urine against bacterial degradation or the post-collectional synthesis of EtG in contaminated samples may cause false interpretation of EtG results in urine samples. This study evaluates the potential of sodium azide in tubes used for urine collection to hinder degradation of ethyl glucuronide by bacterial metabolism taking place during growth of bacterial colonies. The tubes are part of a commercial oral fluid collection device. The sampling system was tested with different gram-positive and gram-negative bacterial species previously observed in urinary tract infections, such as Escherichia coli, Staphylococcus aureus, Enterecoccus faecalis, Staphylococcus epidermidis, Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomonas aeruginosa. Inhibition of bacterial growth by sodium azide, resulting in lower numbers of colony forming units compared to control samples, was observed for all tested bacterial species. To test the prevention of EtG degradation by the predominant pathogen in urinary tract infection, sterile-filtered urine and deficient medium were spiked with EtG, and inoculated with E. coli prior to incubation for 4 days at 37 °C in tubes with and without sodium azide. Samples were collected every 24 hours, during four consecutive days, whereby the colony forming units (CFU) were counted on Columbia blood agar plates, and EtG was analyzed by LC-MS/MS. As expected, EtG degradation was observed when standard polypropylene tubes were used for the storage of contaminated samples. However, urine specimens collected in sodium azide tubes showed no or very limited bacterial growth and no EtG degradation. As a conclusion, sodium azide is useful to reduce bacterial growth of gram-negative and gram-positive bacteria. It inhibits the degradation of EtG by E. coli and can be used for the stabilization of EtG in urine samples.

A Randomized Controlled Trial of Ethyl Glucuronide-Based Contingency Management for Outpatients With Co-Occurring Alcohol Use Disorders and Serious Mental Illness.

OBJECTIVE: The authors examined whether a contingency management intervention using the ethyl glucuronide (EtG) alcohol biomarker resulted in increased alcohol abstinence in outpatients with co-occurring serious mental illnesses. Secondary objectives were to determine whether contingency management was associated with changes in heavy drinking, treatment attendance, drug use, cigarette smoking, psychiatric symptoms, and HIV-risk behavior. METHOD: Seventy-nine (37% female, 44% nonwhite) outpatients with serious mental illness and alcohol dependence receiving treatment as usual completed a 4-week observation period and were randomly assigned to 12 weeks of contingency management for EtG-negative urine samples and addiction treatment attendance, or reinforcement only for study participation. Contingency management included the variable magnitude of reinforcement "prize draw" procedure contingent on EtG-negative samples (<150 ng/mL) three times a week and weekly gift cards for outpatient treatment attendance. Urine EtG, drug test, and self-report outcomes were assessed during the 12-week intervention and 3-month follow-up periods. RESULTS: Contingency management participants were 3.1 times (95% CI=2.2-4.5) more likely to submit an EtG-negative urine test during the 12-week intervention period, attaining nearly 1.5 weeks of additional alcohol abstinence compared with controls. Contingency management participants had significantly lower mean EtG levels, reported less drinking and fewer heavy drinking episodes, and were more likely to submit stimulant-negative urine and smoking-negative breath samples, compared with controls. Differences in self-reported alcohol use were maintained at the 3-month follow-up. CONCLUSIONS: This is the first randomized trial utilizing an accurate and validated biomarker (EtG) to demonstrate the efficacy of contingency management for alcohol dependence in outpatients with serious mental illness.

Predictors of polycyclic aromatic hydrocarbon exposure and internal dose in inner city Baltimore children.

Polycyclic aromatic hydrocarbons (PAHs), the by-products of incomplete combustion of organic materials, are commonly found on particulate matter (PM) and have been associated with the development of asthma and asthma exacerbation in urban populations. We examined time spent in the home and outdoors as predictors of exposures to airborne PAHs and measured urinary 1-hydroxypyrene-glucuronide (1-OHPG) as internal dose of PAHs in 118 children aged 5-12 years from Baltimore, MD. During weeklong periods (Saturday-Saturday) in each of four seasons: daily activities were assessed using questionnaires, indoor air nicotine and PM concentrations were monitored, and urine specimens were collected on Tuesday (day 3) and Saturday (day 7) for measurement of 1-OHPG. Time spent in non-smoking homes was associated with significantly decreased 1-OHPG concentration in urine (ß=-0.045, 95% CI (-0.076, -0.013)), and secondhand smoke (SHS) exposures modified these associations, with higher urinary 1-OHPG concentrations in children spending time in smoking homes than non-smoking homes (P-value for interaction=0.012). Time spent outdoors was associated with increased urinary 1-OHPG concentrations (ß=0.097, 95% CI (0.037, 0.157)) in boys only. Our results suggest that SHS and ambient (outdoor) air pollution contribute to internal dose of PAHs in inner city children.

A simple analytical method of determining 1-hydroxypyrene glucuronide in human urine by isotope dilution with ultra performance liquid chromatography-tandem mass spectrometry.

The glucuronide conjugate of 1-hydroxypyrene (1-OHP-G) is a sensitive and reliable index biomarker for assessing low exposure to polycyclic aromatic hydrocarbons (PAHs). A simple method for determining 1-OHP-G in human urine with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established and applied to evaluate the exposure level of PAHs of pregnant women in a large sample size. After the urine sample was extracted with ethyl acetate, 0.2 mL of the aqueous phase was diluted to 1.0 mL with 5 mmol/L ammonium acetate before injection. The chromatographic separation was performed on a C18 column with a gradient elution and identification was conducted on a tandem mass spectrometry with electrospray ionization in negative mode. 1-OHP-d9-G was used as an internal standard to improve precision. The method was validated and good linearity was obtained in the range of 0.1∼2.0 ng/mL. The limit of detection (LOD) and the limit of quantification (LOQ) of 1-OHP-G were 0.015 and 0.051 ng/mL. Intra-day and inter-day precision were 4.3 and 6.7 %, respectively. The spiked recoveries were 79.4∼106 % for urine samples. This method was rapid, sensitive, and very suitable for batch analysis of urine. Six hundred seventy-five urine samples of pregnant women from the cities of Fuzhou, Shenzhen, and Nanning of P.R. China were analyzed with the proposed method. The medians of 1-OHP-G concentration were 0.27 µg/g.cr (n = 201), 0.30 µg/g.cr (n = 212), and 0.51 µg/g.cr (n = 262) for the cities of Fuzhou, Shenzhen, and Nanning, respectively. 1-OHP-G concentrations in urine samples of pregnant women from the cities of Fuzhou and Shenzhen in coastal areas were both significantly lower than that of Nanning City in inland region (p < 0.001). Graphical Abstract The internal standard 1-OHP-d9-G and 2.0 mL of ethyl acetate were added to 1.0 mL of urine sample, after vortex vibration and centrifugation the aqueous phase was removed and diluted, and 5 µl of aliquot was analyzed by UPLC-MS/MS.

Excessive urinary excretion of isopropyl glucuronide after isopropanol abuse.

BACKGROUND: Ethyl glucuronide (EtG) in urine is considered a marker of alcohol consumption. We present a case of a false-positive immunological EtG screening result due to excessive isopropyl glucuronide excretion in urine of an alcohol-dependent patient with a history of industrial cleaning fluid abuse. METHODS: EtG screening was done with the Microgenics DRI EtG enzyme immunoassay on a Beckman Coulter AU680 analyzer according to the testkit instructions. Confirmatory analysis was done by LC-MS/MS for EtG, 1-propyl (syn. n-propyl), 2-propyl (syn. isopropyl), 1-butyl, 2-butyl, and tert-butyl glucuronide. Both methods were validated according to the Guidelines of the Society of Toxicological and Forensic Chemistry (GTFCh, Germany). RESULTS: EtG screening by immunoassay was positive, approx. 860mg/L or approx. 1540mg/g creatinine (forensic cut-off 0.1mg/L, clinical cut-off 0.5mg/L). LC-MS/MS confirmatory analysis was negative for EtG (<0.05mg/L; forensic cut-off 0.1mg/L), but strongly positive for 2-propyl glucuronide (approx. 1100mg/L or 2000mg/g creatinine; cut-off 0.1mg/L). 1-propyl, 1-butyl, and tert-butyl glucuronide were negative (<0.05mg/L; cut-off 0.1mg/L), 2-butyl glucuronide was 0.1mg/L (cut-off 0.1mg/L). CONCLUSION: Consumption of household and industrial chemicals with short chain aliphatic alcohols should be considered a rare but potential source of false-positive EtG immunoassay results. Glucuronides from frequently used short chain aliphatic alcohols, like 1-propanol (syn. n-propanol) and 2-propanol (syn. isopropanol) as the most important disinfectant components, should be included into EtG confirmatory analysis. This will be helpful not only for the assessment of the source for remarkable EtG immunoassay results, it can also contribute to a more specific diagnosis in cases with suspected intoxication by consumer or industrial chemical products. Excessive urinary 2-propyl glucuronide (syn. isopropyl glucuronide) concentrations should be considered a marker of isopropanol intoxication.

Association between Glucuronidation Genotypes and Urinary NNAL Metabolic Phenotypes in Smokers.

BACKGROUND: The most abundant and potent carcinogenic tobacco-specific nitrosamine in tobacco and tobacco smoke is 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). In vivo, NNK is rapidly metabolized to both the (R)- and (S)-enantiomers of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), which possesses similar carcinogenic properties as NNK. The major detoxification pathway for both NNAL enantiomers is glucuronidation by UDP-glucuronosyltransferase (UGT) enzymes including UGT2B10 and UGT2B17. The goal of the present study was to directly examine the role of UGT genotypes on urinary levels of NNAL glucuronides in smokers. METHODS: NNAL-N-Gluc, (R)-NNAL-O-Gluc, (S)-NNAL-O-Gluc, and free NNAL were simultaneously and directly quantified in the urine of smokers by LC/MS analysis. Genotypes were determined by TaqMan assay using genomic DNA. RESULTS: The functional knockout polymorphism in the UGT2B10 gene at codon 67 (Asp>Tyr) was significantly (P < 0.0001) associated with a 93% decrease in creatinine-adjusted NNAL-N-Gluc. The polymorphic whole-gene deletion of the UGT2B17 gene was associated with significant (P = 0.0048) decreases in the levels of creatinine-adjusted (R)-NNAL-O-Gluc, with a 32% decrease in the levels of urinary (R)-NNAL-O-Gluc/(S)-NNAL-O-Gluc among subjects with the UGT2B17 (*2/*2) genotype as compared to subjects with the UGT2B17 (*1/*1) genotype. CONCLUSIONS: These results suggest that functional polymorphisms in UGT2B10 and UGT2B17 are associated with a reduced detoxification capacity against NNAL and may therefore affect individual cancer risk upon exposure to tobacco. IMPACT: This is the first report to clearly demonstrate strong genotype-phenotype associations between both the UGT2B10 codon 67 Asp

The effects of alcohol-containing e-cigarettes on young adult smokers.

BACKGROUND: The liquids (e-liquids) used in an electronic cigarette (e-cigarette) contain myriad chemicals without adequate human inhalation safety data. Furthermore, the absence of e-liquid labeling requirements poses a formidable challenge to understanding how e-liquid constituents may promote nicotine addiction and/or have independent or synergistic biological effects when combined with nicotine. Ethyl alcohol is such a constituent, but has received little scientific interest in this context. METHODS: Using a randomized, double blind, crossover design, acute changes in subjective drug effects, motor performance and biochemical measures of alcohol and nicotine intake were evaluated after directed and ad lib puffing from two commercially available e-liquids containing nicotine (8 mg/ml), vanilla flavor and either 23.5% (high) or 0.4% (trace) alcohol. RESULTS: While no differences in subjective drug effects were observed between alcohol conditions, performance on the Purdue Pegboard Dexterity Test (PPDT) improved under the trace, but not under the 23.5% alcohol condition. Although plasma alcohol levels remained undetectable during testing, urine ethyl glucuronide (EtG), an alcohol metabolite, became measurable in three participants after puffing from the 23.5% alcohol e-cigarette. CONCLUSIONS: Brief use of a widely available type of e-cigarette containing an e-liquid purchased from an internet vendor can negatively impact psychomotor performance and in some instances, produce detectable levels of a urine alcohol metabolite. Given the widespread and unregulated use of e-cigarettes, especially by youth and other vulnerable populations, further studies are needed to evaluate both the acute safety and long-term health risks of using alcohol-containing e-cigarettes.

A culturally-tailored behavioral intervention trial for alcohol use disorders in three American Indian communities: Rationale, design, and methods.

BACKGROUND: Disproportionately high rates of alcohol use disorders are present in many American Indian/Alaska Native (AI/AN) communities, yet little information exists regarding the effectiveness of alcohol treatments in AI/AN populations. Contingency management is an intervention for illicit drug use in which tangible reinforcers (rewards) are provided when patients demonstrate abstinence as assessed by urine drug tests. Contingency management has not been widely studied as an intervention for alcohol problems because until recently, no alcohol biomarker has been available to adequately verify abstinence. AIMS: The HONOR Study is designed to determine whether a culturally-tailored contingency management intervention is an effective intervention for AI/AN adults who suffer from alcohol use disorders. METHODS: Participants include 400 AI/AN alcohol-dependent adults residing in one rural reservation, one urban community, as well as a third site to be decided, in the Western U.S. Participants complete a 4-week lead-in phase prior to randomization, then 12 weeks of either a contingency management intervention for alcohol abstinence, or a control condition where participants receive reinforcers for attending study visits regardless of alcohol use. Participants are then followed for 3-more months post-intervention. The primary study outcome is urinary ethyl glucuronide-confirmed alcohol abstinence; secondary outcomes include self-reported alcohol and drug use, HIV risk behaviors, and self-reported cigarette smoking. DISCUSSION: This will be the largest randomized, controlled trial of any alcohol for AI/ANs and the largest contingency management study targeting alcohol use disorders, thus providing important information to AI/AN communities and the alcohol treatment field in general.

Quantitation of ethyl glucuronide in serum & urine by gas chromatography - mass spectrometry.

BACKGROUND & OBJECTIVES: Alcohol misuse has now become a serious public health problem and early intervention is important in minimizing the harm. Biochemical markers of recent and high levels of alcohol consumption can play an important role in providing feedback regarding the health consequences of alcohol misuse. Existing markers are not sensitive to recent consumption and in detecting early relapse. Ethyl glucuronide (EtG), a phase-II metabolite of ethanol is a promising marker of recent alcohol use and can be detected in body fluids. In this study an analytical technique for quantitation of EtG in body fluids using solid-phase extraction (SPE) and gas chromatography (GC) with mass spectrometric detection (MS) was developed and validated. METHODS: De-proteinization of serum and urine samples was done with perchloric acid and hydrochloric acid, respectively. Serum samples were passed through phospholipids removal cartridges for further clean up. EtG was isolated using amino propyl solid phase extraction columns. Chromatographic separation was achieved by gas chromatography with mass spectrometry. RESULTS: Limit of detection and limit of quantitation were 50 and 150 ng/ml for urine and 80 and 210 ng/ml for serum, respectively. Signal to noise ratio was 3:1, mean absolute recovery was 80-85 per cent. Significant correlation was obtained between breath alcohol and serum EtG levels (r=0.853) and urine EtG and time since last abuse (r = -0.903) in clinical samples. INTERPRETATION & CONCLUSIONS: In the absence of other standardized techniques to quantitate EtG in biological samples, this gc - ms method was found to have high throughput and was sensitive and specific.

Levels of tobacco-specific metabolites among non-smoking lung cancer cases at diagnosis: case-control findings.

BACKGROUND: Environmental tobacco smoking (ETS) significantly contributes to morbidity and mortality and is a known risk factor for lung cancer development in lifelong nonsmokers. The metabolite 4-(methylnitrosamino)- 1-(3-pyridyl)-1-butanol (NNAL) and its glucuronides (NNAL-Glucs) have now emerged as leading biomarkers for the study of carcinogen exposure in non-smokers exposed to ETS. MATERIALS AND METHODS: We carried out our study on NNAL in the urine of non-smokers exposed to ETS and the association between ETS and lung cancer. Subjects were enrolled from 2008-2010. NNAL was analyzed for 74 non-smoking lung cancer and 85 healthy controls. The main objective of this study was to provide an estimate of the risk of lung cancer from exposure to ETS in the Korean population. RESULTS: The mean NNAL concentration in urine was significantly lower in non-smoking patient groups (n=74) than in control groups (n=85) (4.7±15.0 pg/mg, 6.5 ± 17.9 pg/mg, respectively, Mann-Whitney U test, p<0.001). CONCLUSIONS: The urine NNAL of non-smoking patients with lung cancer was not elevated with regard to the non-smoking control group. This may be due to life-style changes after diagnosis. A prospective study will be needed to evaluate the association of NNAL and non-smoking lung cancer.

Assessing traffic and polycyclic aromatic hydrocarbon exposure in Montreal, Canada.

INTRODUCTION: The International Agency for Research on Cancer classifies specific polycyclic aromatic hydrocarbons (PAHs) as probable carcinogens. This study compares two PAH biomarkers and their relationship with geographic information system (GIS) based traffic density (a proxy of PAH exposure), and explores the determinants of the PAH biomarkers. METHODS: A cross-sectional study was conducted in Montreal with 200 volunteers (107 females and 93 males) ages 20 to 53 years. Data were collected by questionnaire, urine samples were used for biomarker analysis, and innovative GIS-based time- and distance-weighted traffic densities (TDWTD) were calculated for all locations of participants during the 48 h prior to urine collection. RESULTS: Detection rates of the two biomarkers were greater than 95%. Female participants had higher 1-OHP and 1-OHPG levels than males, and no relationship was detected between TDWTD in 48 h and the two PAH biomarkers. Biomarker levels were related to smoking more than one pack of cigarettes in the previous 48 h, and among non-smokers, barbecued meat consumption increased the level of urinary 1-OHP (exp ß: 1.45, 95% CI: 1.07 to 1.98). CONCLUSIONS: Both 1-OHP and 1-OHPG can be used to assess the relatively low PAH levels to which the general population is exposed. With the exception of smoking, the impact of PAH exposure factors on the biomarkers is relatively small in this study population.

Development of a rapid method for the simultaneous separation and determination of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and its N- and O-glucuronides in human urine by liquid chromatography-tandem mass spectrometry.

Determination of the tobacco-specific nitrosamine metabolite 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and its N- and O-glucuronides (NNAL-N-Gluc and NNAL-O-Gluc) is important for toxicology analysis of tobacco smoke induced carcinogenicity and the understanding of detoxification mechanisms of the carcinogenic nitrosamine in humans. But previously reported indirect measurement methods involving enzymolysis and base treatment steps were tedious and time-consuming. In this work, a direct measurement method for simultaneous determination of urinary NNAL, NNAL-N-Gluc and NNAL-O-Gluc by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in a single run was developed for the first time without the need to perform enzymatic or base hydrolysis. Urine samples were purified using dichloromethane and further extracted by solid-phase extraction. Then they were analyzed by LC-MS/MS operated in electrospray positive ionization mode. Chromatographic separation was achieved on a Phenomenex Kinetex PFP column within 6 min. The proposed method was validated and the results demonstrated that the method can produce satisfactory recoveries and reproducibility for the analytes. The applicability of this newly developed method was investigated for the simultaneous analysis of the three metabolites in smokers' urine and the obtained results were comparable to those detected using the conventional enzymolysis method.