Driving license regranting: Hair EtG, serum CDT, and the role of sociodemographic and medicolegal variables.

Driving under the influence (DUI) of alcohol is a road safety problem. Driving license regranting is based on the evaluation of medicolegal and toxicological variables that may include serum carbohydrate-deficient transferrin (CDT) and hair ethyl glucuronide (hEtG). The aim of the study was to compare the diagnostic performance of CDT and hEtG in a population of DUI offenders. Other factors potentially associated with heavy alcohol use were explored. The population included DUI offenders examined during the period of January 1, 2019, through June 30, 2022. Sociodemographic, medicolegal, and toxicological variables were collected. CDT in serum and EtG in head hair were determined in all subjects. Excessive alcohol intake (hEtG ≥30 pg/mg) was considered cause for unfitness to drive. Cohen's kappa coefficient was calculated. Descriptive analyses were performed using chi-square and Mann-Whitney tests. Variables significantly different between the groups were included in a multivariate binary logistic regression model. The sample encompassed 838 subjects (case group: 179, comparison group: 689). CDT exhibited poor agreement (κ = 0.053) with hEtG as the reference test. Lower education, age at DUI, heavy smoking, and GGT levels associated with heavy alcohol consumption differentiated the two groups. For DUI offenders, the use of CDT to assess heavy alcohol consumption is limited, possibly due to the time-window assessed, the time required for normalization, and the different amount of ethanol needed to reach higher CDT levels, in comparison to hEtG; thus, hEtG assessment is strongly recommended for this population. Heavy smoking, GGT, education, and age could be related to heavy alcohol consumption and higher risk of DUI.

The Contents of Polyphenols in Perilla frutescens (L.) Britton var. frutescens (Egoma) Leaves are Determined by Vegetative Stage, Spatial Leaf Position, and Timing of Harvesting during the Day.

The leaf of Perilla frutescens (L.) Britton var. frutescens (egoma) is a rich source of polyphenolic compounds, including rosmarinic acid. However, there is still a lack of detailed information concerning the content of phenolic compounds in these leaves. Since some flavonoids were found as a conjugated form, leaves were used untreated or hydrolyzed using ß-glucuronidase for analysis. Enzymatic hydrolysis method successfully identified some polyphenols, which have not been reported before. Scutellarin, a flavone glucuronide with a molecular mass similar to that of luteolin 7-O-glucuronide, was present in egoma leaves. Scutellarin was the second most abundant polyphenolic compound, after rosmarinic acid. Egoma leaves at the top of the plant contained a higher amount of rosmarinic acid and scutellarin compared to that in the leaves below. The difference in plant growth stage also influenced the rosmarinic acid and scutellarin contents, while the time of harvesting during the day did rosmarinic acid contents only. This is the first time that scutellarin, a traditional Chinese medicine, widely used for the treatment of cerebrovascular disease, was quantitatively determined in egoma leaves. The present study may help adding value to egoma leaves, developing dietary supplements, functional foods, and cosmetics.

Exploring extra dimensions to capture saliva metabolite fingerprints from metabolically healthy and unhealthy obese patients by comprehensive two-dimensional gas chromatography featuring Tandem Ionization mass spectrometry.

This study examines the information potential of comprehensive two-dimensional gas chromatography combined with time-of-flight mass spectrometry (GC×GC-TOF MS) and variable ionization energy (i.e., Tandem Ionization™) to study changes in saliva metabolic signatures from a small group of obese individuals. The study presents a proof of concept for an effective exploitation of the complementary nature of tandem ionization data. Samples are taken from two sub-populations of severely obese (BMI > 40 kg/m2) patients, named metabolically healthy obese (MHO) and metabolically unhealthy obese (MUO). Untargeted fingerprinting, based on pattern recognition by template matching, is applied on single data streams and on fused data, obtained by combining raw signals from the two ionization energies (12 and 70 eV). Results indicate that at lower energy (i.e., 12 eV), the total signal intensity is one order of magnitude lower compared to the reference signal at 70 eV, but the ranges of variations for 2D peak responses is larger, extending the dynamic range. Fused data combine benefits from 70 eV and 12 eV resulting in more comprehensive coverage by sample fingerprints. Multivariate statistics, principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA) show quite good patient clustering, with total explained variance by the first two principal components (PCs) that increases from 54% at 70 eV to 59% at 12 eV and up to 71% for fused data. With PLS-DA, discriminant components are highlighted and putatively identified by comparing retention data and 70 eV spectral signatures. Within the most informative analytes, lactose is present in higher relative amount in saliva from MHO patients, whereas N-acetyl-D-glucosamine, urea, glucuronic acid γ-lactone, 2-deoxyribose, N-acetylneuraminic acid methyl ester, and 5-aminovaleric acid are more abundant in MUO patients. Visual feature fingerprinting is combined with pattern recognition algorithms to highlight metabolite variations between composite per-class images obtained by combining raw data from individuals belonging to different classes, i.e., MUO vs. MHO.Graphical abstract.

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

INTRODUCTION: In 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. AIMS: The aim of the present study was to assess the suitability of assaying iron concentrations in liver and other tissues as postmortem alcoholism marker. METHODS: The 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. RESULTS: No 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. CONCLUSION: The 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.

Risk factors associated to tobacco and alcohol use in a large French cohort of pregnant women.

Tobacco and/or alcohol use during pregnancy is a major public health concern. The aim of our study was to identify risk factors associated to maternal alcohol and tobacco use assessed by maternal self-reports combined with biological measurements in meconium samples of cotinine and ethylglucuronide which reflect fetal exposure to tobacco and alcohol, respectively, during the 3rd trimester of pregnancy. We conducted a prospective study in three maternity hospitals in a large urban area during consecutive weeks (2010 and 2011). Maternal sociodemographic and clinical characteristics were assessed after delivery, using the French version of the Addiction Severity Index. Cotinine and ethylglucuronide were measured in meconium samples. Seven hundred and twenty-four women were included, and 645 meconium samples collected. Using multivariate analyses, we found that not being married or having a smoking partner predicts maternal tobacco use. In contrast, a decreased risk was associated with higher education level and wanted pregnancy. The risk for alcohol use increased when the mother had been in conflict with any relative or her partner for a long time throughout her life, as well as in case of previous treatment for any mental or emotional disorder. Using multivariate analyses and cotinine presence in meconium samples, the risks were similar except for marital status, which was not associated to cotinine presence. Community education and prevention programs should urgently be improved for all women of childbearing age with a special focus on those with past histories of mental or emotional disorders and addictive disorders. Smoking cessation should be recommended to both parents.

Differential correlations between maternal hair levels of tobacco and alcohol with fetal growth restriction clinical subtypes.

Maternal exposure to tobacco and alcohol is a known cause, among others, for fetal growth restriction (FGR). Clinically, FGR can be subclassified into two forms: intrauterine growth restriction (IUGR) and small for gestational age (SGA), based on the severity of the growth retardation, and abnormal uterine artery Doppler or cerebro-placental ratio. This study aimed at investigating any differential correlation between maternal exposures to these toxins with the two clinical forms of FGR. Therefore, a case-control study was conducted in Barcelona, Spain. Sixty-four FGR subjects, who were further subclassified into IUGR (n = 36) and SGA (n = 28), and 89 subjects matched appropriate-for-gestational age (AGA), were included. The levels of nicotine (NIC) and ethyl glucuronide (EtG), biomarkers of tobacco and alcohol exposure, respectively, were assessed in the maternal hair in the third trimester. Our analysis showed 65% of the pregnant women consumed alcohol, 25% smoked, and 19% did both. The odds ratios (ORs) of IUGR were 21 times versus 14 times for being SGA with maternal heavy smoking, while with alcohol consumption the ORs for IUGR were 22 times versus 37 times for the SGA group. The differential correlations between these toxins with the two subtypes of FGR suggest different mechanisms influencing fetal weight. Our alarming data of alcohol consumption during pregnancy should be considered for further confirmation among Spanish women.

Stability of alcohol and tobacco consumption biomarkers in a real rising main sewer.

Since alcohol and tobacco consumption are among the leading causes of population health harm, it is very important to understand the consumption behaviour to develop effective harm reduction strategies. Wastewater-based epidemiology (WBE) is a potential tool for estimating their consumption, but there are several uncertainties that need to be determined, including the stability of biomarkers in the sewer. Utilizing a real rising main sewer, this study investigated the stability of alcohol and tobacco consumption biomarkers. Rhodamine and acesulfame were used as flow tracer and benchmarker to understand the transportation of wastewater in the sewer with a hydraulic retention time between 2.7 and 5.0 h. Ethyl sulphate (EtS) and ethyl glucuronide (EtG), two biomarkers of alcohol consumption, were found to have different in-sewer stability, with EtS much more stable than EtG. The degradation rate of EtS is approximately 8% per hour, while EtG has a half-life of 1.9 h. Formation of nicotine, cotinine and trans-3'-hydroxycotinine, three biomarkers for tobacco consumption, was observed during the experiment, probably due to deconjugation of their glucuronide chemicals. The deconjugation process has prevented the determination of actual stability of the three chemicals. However, it is suggested that cotinine is relatively stable, while nicotine and trans-3'-hydroxycotinine degrade to a certain degree in the sewer system. According to our findings, the in-sewer degradation is more important during the interpretation of alcohol consumption estimation than for tobacco consumption estimation.

Different temperatures select distinctive acetic acid bacteria species and promotes organic acids production during Kombucha tea fermentation.

Kombucha is a traditional beverage produced by tea fermentation, carried out by a symbiotic consortium of bacteria and yeasts. Acetic Acid Bacteria (AAB) usually dominate the bacterial community of Kombucha, driving the fermentative process. The consumption of this beverage was often associated to beneficial effects for the health, due to its antioxidant and detoxifying properties. We characterized bacterial populations of Kombucha tea fermented at 20 or 30 °C by using culture-dependent and -independent methods and monitored the concentration of gluconic and glucuronic acids, as well as of total polyphenols. We found significant differences in the microbiota at the two temperatures. Moreover, different species of Gluconacetobacter were selected, leading to a differential abundance of gluconic and glucuronic acids.

Potential impact of the sewer system on the applicability of alcohol and tobacco biomarkers in wastewater-based epidemiology.

Understanding the actual consumption of alcohol and tobacco in the population is important for forming public health policy. For this purpose, wastewater-based epidemiology has been applied as a complementary method to estimate the overall alcohol and tobacco consumption in different communities. However, the stability of their consumption biomarkers - ethyl sulfate, ethyl glucuronide, cotinine, and trans-3'-hydroxycotinine - in the sewer system has not yet been assessed. This study aimed to conduct such assessment using sewer reactors mimicking conditions of rising main, gravity sewer, and wastewater alone, over a 12-hour period. The results show that cotinine and trans-3'-hydroxycotinine are relatively stable under all sewer conditions while ethyl sulfate was only stable in wastewater alone and gradually degraded in rising main and gravity sewer conditions. Ethyl glucuronide quickly degraded in all reactors. These findings suggest that cotinine and trans-3'-hydroxycotinine are good biomarkers to estimate tobacco consumption; ethyl sulfate may be used as a biomarker to estimate alcohol consumption, but its in-sewer loss should be accounted for in the calculation of consumption estimates. Ethyl glucuronide, and probably most of glucuronide compounds, are not suitable biomarkers to be used in wastewater-based epidemiology due to their in-sewer instability.

Environmental-friendly technologies for the production of antioxidant xylooligosaccharides from wheat chaff.

Ultrasound irradiation and enzymatic hydrolysis were applied for the production of antioxidant xylooligosaccharides from wheat chaff. To facilitate enzyme action, 3% (w/w) suspension of raw material was pretreated by ultrasound at 25Hz, 540W for 10min. Ultrasound treatment released xylooligosaccharides with predominant xylotriose into liquid fraction which expressed 1.03±0.01 (µmolAAE/g) ABTS radicals-scavenging activity. Endo-xylanase action on pretreated wheat chaff released 21.76±1.42 and 32.3±0.75mg reducing sugars equivalents/g after 24h when applied at dosages 0.15 and 0.3U/g. respectively. With increase in reaction time portion of xylotriose and xylotetraose in hydrolysates was increase relatively to xylopentose as well as quantities of xylose and glucose. Extremely significant inverse correlation between monosaccharides/oligosaccharides ratio and ABTS radical-scavenging activity was determined. Results indicated potential of environmental friendly ultrasound and enzymatic technologies for the production of xylooligosaccharides from wheat chaff for potential food application.

Alcohol, caffeine, and nicotine consumption in adolescents: hair analysis versus self-report.

BACKGROUND: Clinical reliability of self-reported data for alcohol, caffeine, and nicotine consumptions is lacking, particularly in adolescents. OBJECTIVES: To compare a self-report questionnaire and hair analysis to assess the reliability and effectiveness of the self-report. METHODS: A cross-sectional study on 14-15-year-old Italian students (n = 874, 38% males, 62% females) was performed comparing self-reported data to hair analysis. The latter quantified hair concentrations of caffeine, nicotine, cotinine, ethyl glucuronide (EtG), and fatty acid ethyl esters (FAEEs) using mass spectrometry. RESULTS: Concordance between self-report and hair testing ranged from good to poor across substances and levels of use: poor for heavy alcohol intake (EtG: k = 0.36, 20 positive cases by hair analysis, false negative by self-report, 2.3% of total sample; FAEE k = 0.31, 25 positive cases, 2.9% of total sample); fair to poor for active smokers (k = 0.40, 125 positive cases, 14.3% of total sample); and moderate for caffeine (k = 0.57, 56 positive cases, 6.4% of total sample). CONCLUSIONS: Epidemiological studies on alcohol, caffeine, and nicotine consumption in adolescents may benefit from the inclusion of toxicological analysis on hair samples to overcome the under-reporting phenomenon of questionnaires and detect more cases of problematic substance use.

Alcohol Consumption during Pregnancy: Analysis of Two Direct Metabolites of Ethanol in Meconium.

Alcohol consumption in young women is a widespread habit that may continue during pregnancy and induce alterations in the fetus. We aimed to characterize prevalence of alcohol consumption in parturient women and to assess fetal ethanol exposure in their newborns by analyzing two direct metabolites of ethanol in meconium. This is a cross-sectional study performed in September 2011 and March 2012 in a series of women admitted to an obstetric unit following childbirth. During admission, socio-demographic and substance use (alcohol, tobacco, cannabis, cocaine, and opiates) during pregnancy were assessed using a structured questionnaire and clinical charts. We also recorded the characteristics of pregnancy, childbirth, and neonates. The meconium analysis was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to detect the presence of ethyl glucuronide (EtG) and ethyl sulfate (EtS). Fifty-one parturient and 52 neonates were included and 48 meconium samples were suitable for EtG and EtS detection. The median age of women was 30 years (interquartile range (IQR): 26-34 years); EtG was present in all meconium samples and median concentration of EtG was 67.9 ng/g (IQR: 36.0-110.6 ng/g). With respect to EtS, it was undetectable (<0.01 ng/g) in the majority of samples (79.1%). Only three (6%) women reported alcohol consumption during pregnancy in face-to-face interviews. However, prevalence of fetal exposure to alcohol through the detection of EtG and EtS was 4.2% and 16.7%, respectively. Prevention of alcohol consumption during pregnancy and the detection of substance use with markers of fetal exposure are essential components of maternal and child health.

Gas chromatographic determination of ethyl glucuronide in hair: comparison between tandem mass spectrometry and single quadrupole mass spectrometry.

Ethyl glucuronide (EtG), a minor metabolite of ethanol, accumulates in hair and is currently used as a long-term marker for the detection of chronic and excessive alcohol consumption. Sensitive methods are required to differentiate teetotalers from moderate drinkers according to the established cut-off (i.e., 7 pg/mg hair). The aim of this study was to develop a sensitive method using gas chromatography coupled to tandem mass spectrometry (GC-MS/MS) operated in the negative ion chemical ionization (NICI) mode. The validated method was applied to hair samples from teetotalers, moderate and excessive alcohol consumers, and results were compared to a previously validated GC-NICI-MS method. The developed GC-NICI-MS/MS method showed linearity over a range from 2 to 400 pg/mg hair, with a limit of detection (LOD) of 0.05 pg/mg hair and a lower limit of quantification (LLOQ) of 0.2 pg/mg hair, compared to an LOD of 0.5 pg/mg hair and LLOQ of 1.5 pg/mg hair obtained with GC-NICI-MS. Furthermore, lower background noise was observed using GC-NICI-MS/MS. Comparison of results of hair samples (n=58) obtained by GC-NICI-MS and GC-NICI-MS/MS showed no significant difference between both methods (paired-sample t-test, p>0.05; mean CV=1.0%). The differences between both methods were larger for EtG concentrations<30 mg/pg hair (mean CV=1.7%) than for EtG concentrations>30 mg/pg hair (mean CV=0.7%). This suggests a higher selectivity of GC-NICI-MS/MS at lower concentrations. In conclusion, by using GC-NICI-MS/MS, a higher analytical selectivity and an improved signal to noise ratio, can be achieved. Although GC-NICI-MS would not change the interpretation of the EtG concentrations, the present GC-NICI-MS/MS method should preferentially be used for the determination of EtG in hair, especially when differentiating between teetotalers and moderate drinkers according to the current cut-off (i.e., 7 pg/mg hair).

Burnout syndrome among undergraduate nursing students at a public university / Síndrome de Burnout entre estudantes de graduação em enfermagem de uma universidade pública / Síndrome de Burnout entre estudiantes de pregrado en enfermería de una universidad pública

OBJECTIVE: to investigate the burnout syndrome and its relationship with demographic and academic variables among undergraduate nursing students at a public university in Southern Brazil. METHOD: a quantitative study with 168 students, by applying an adaptation of the Maslach Burnout Inventory - Student Survey, validated for this study. We used descriptive and variance analysis of the data analysis. RESULTS: we found that students do not have the burnout syndrome, manifesting high average scores in Emotional Exhaustion, low in Disbelief and high in Professional Effectiveness; that younger students who perform leisure activities have greater Professional Effectiveness, unlike students in early grades with no extracurricular activities; combining work and studies negatively influenced only the Professional Effectiveness factor, while the intention of giving up influenced negatively Disbelief and Professional Effectiveness factors. CONCLUSION: the situations that lead students to Emotional Exhaustion need to be recognized, considering the specificity of their study environments. .

OBJETIVO: investigar a síndrome de Burnout e sua relação com variáveis sociodemográficas e acadêmicas, entre estudantes de graduação em enfermagem de uma universidade pública do Sul do Brasil. MÉTODO: estudo quantitativo, realizado com 168 estudantes, mediante a aplicação de uma adaptação do Maslach Burnout Inventory - Student Survey, validada para este estudo. Utilizou-se a análise descritiva e de variância para análise dos dados. RESULTADOS: constatou-se que os estudantes não apresentam a síndrome de Burnout, manifestando médias altas em exaustão emocional, baixas em descrença e altas em eficácia profissional; que estudantes mais jovens e que realizam atividades de lazer apresentam maior eficácia profissional, diferentemente de estudantes das séries iniciais e que não realizam atividades extracurriculares; conciliar trabalho e estudos influenciou negativamente apenas o fator eficácia profissional, enquanto a intenção de desistir do curso influenciou negativamente os fatores descrença e eficácia profissional. CONCLUSÃO: faz-se necessário o reconhecimento das situações que levam os estudantes à exaustão emocional, considerando a especificidade de seus ambientes de formação. .

OBJETIVO: investigar la síndrome de burnout y su relación con variables sociodemográficas y académicas, entre estudiantes de pregrado en enfermería de una universidad pública del Sur de Brasil. MÉTODO: estudio cuantitativo, desarrollado con 168 estudiantes, mediante la aplicación de una adaptación del Maslach Burnout Inventory - Student Survey, validada para fines de ese estudio. Fueron utilizados los análisis descriptivo y de variancia para analizar los datos. RESULTADOS: se constató que los estudiantes no presentan la síndrome de burnout, manifestando altos promedios en Agotamiento Emocional, bajos en Descreencia y altos en Eficacia Profesional; que estudiantes más jóvenes y que practican actividades de ocio presentan mayor Eficacia Profesional, diferentemente de estudiantes de los años iniciales sin actividades extracurriculares; conciliar trabajo y estudios influyó negativamente apenas el factor Eficacia Profesional, mientras la intención de desistir del curso influyó negativamente los factores Descreencia y Eficacia Profesional. CONCLUSIÓN: es necesario reconocer las situaciones que llevan a los estudiantes al Agotamiento Emocional, considerando la especificidad de sus ambientes de formación. .

Degradation of the ethyl glucuronide content in hair by hydrogen peroxide and a non-destructive assay for oxidative hair treatment using infra-red spectroscopy.

The assessment of quantification results of the alcohol abuse marker ethyl glucuronide (EtG) in hair in comparison to the cut-off values for the drinking behavior may be complicated by cosmetic hair bleaching. Thus, the impact of increasing exposure to hydrogen peroxide on the EtG content of hair was investigated. Simultaneously, the change of absorbance in the range of 1000-1100 cm(-1) indicative for the oxidation of cystine was investigated non-destructively by attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) using pulverized portions of the respective hair samples. Hair samples treated with hydrogen peroxide consistently displayed a significantly increased absorbance at 1040 cm(-1) associated with the formation of cysteic acid. The EtG content decreased significantly if the hair was treated with alkaline hydrogen peroxide as during cosmetic bleaching. It could be shown that ATR-FTIR is capable of detecting an exposure to hydrogen peroxide when still no brightening was visible and already before the EtG content deteriorated significantly. Thus, hair samples suspected of having been exposed to oxidative treatment may be checked non-destructively by a readily available technique. This assay is also possible retrospectively after EtG extraction and using archived samples.

Application of isoabsorption plots generated by high-performance liquid chromatography with diode array detection to the development of multicomponent quantitative analysis of traditional herbal medicines.

Multicomponent quantitative analysis is one of the mainstream quality control methods of traditional herbal medicines. Since the constituents of traditional herbal medicines samples are complex, the development of high-performance liquid chromatography methods is laborious. In this study, an isoabsorption plot, a chromatographic/spectrometric data image plotted by diode array detection was utilized to facilitate the establishment of a high-performance liquid chromatography method by optimizing and validating the detection conditions off-line. Consequently a simple, reliable and accurate method for simultaneous determination of seven active polyphenolic components (protocatechuic acid, chlorogenic acid, caffeic acid, p-coumaric acid, rosmarinic acid, scutellarin, and apigenin) in Qingfei mixture, a long-used Chinese prescription, was developed. The chromatographic separation was performed on a C18 column with gradient elution of phosphoric acid aqueous solution (0.05%, v/v) and acetonitrile, and a wavelength switch program optimized with isoplot was adopted for detection. The method was validated in terms of linearity, sensitivity, precision, repeatability, and accuracy and was successfully applied to the simultaneous determination of the seven polyphenolic components in different production batches of Qingfei Mixture. These results indicated that isoplot is an effective tool to improve the establishment of multicomponent quantitative analysis methods.

Determination of ethyl glucuronide in hair: a rapid sample pretreatment involving simultaneous milling and extraction.

A combination of simultaneous milling and extraction known as micropulverized extraction was developed for the quantification of the alcohol marker ethyl glucuronide (EtG) in hair samples using a homogeneous reference material and a mixer mill. Best extraction results from 50 mg of hair were obtained with 2-mL plastic tubes containing two steel balls (∅ = 5 mm), 0.5 mL of water and with an oscillating frequency of 30 s(-1) over a period of 30 min. EtG was quantified employing a validated GC-MS procedure involving derivatization with pentafluoropropionic acid anhydride. This micropulverization procedure was compared with dry milling followed by separate aqueous extraction and with aqueous extraction after manual cutting to millimeter-size snippets. Micropulverization yielded 28.0 ± 1.70 pg/mg and was seen to be superior to manually cutting (23.0 ± 0.83 pg/mg) and equivalent to dry grinding (27.7 ± 1.71 pg/mg) with regard to completeness of EtG extraction. The option to process up to 20 samples simultaneously makes micropulverization especially valuable for the high throughput of urgent samples.

A Norwegian study of the suitability of hair samples in epidemiological research of alcohol, nicotine and drug use.

A feasibility study was performed to examine the effectiveness of hair testing in determining the prevalence of drug use in a young adult population. The study included 200 randomly selected young adults in Norway. It was designed to make the collection, preparation and analysis of the samples as little resource demaning as possible. Full anonymity was provided for the participants. In total, 23.5% of the samples were positive for one or more substances (14.5%, excluding the nicotine metabolite cotinine). Of the samples, 5% were positive for at least one illegal drug, 9.5% for a medicinal drug, 11.5% for cotinine and 2.5% for the alcohol metabolite ethyl glucuronide. The preliminary findings suggest that the study protocol used to collect and analyze the samples was unable to produce results that could be generalized to the young adult population in Norway. Analysis of hair samples may underestimate the use of cannabis, alcohol, amphetamine and methamphetamine. It may, however, be done to estimate cocaine and general drug use if a sample-collection procedure different from that described in our study is used and includes information about hair length, sample length, length from the scalp, cosmetic treatment, washing and whether the samples always get washed/decontaminated prior to analysis.

The homogeneity testing of EtG in hair reference materials: a high-throughput procedure using GC-NCI-MS.

The validation of a robust quantification procedure for EtG in hair using GC-NCI-MS is presented. Aqueous extraction is followed by complete lyophylization of the extract and derivatization with pentafluoropropionic anhydride (PFPA) under controlled temperature and duration. Clean-up of extracts was dispensable and standard single quadrupole MS displayed sufficient selectivity and sensitivity. The method displayed a wide linearity range and enabled LOD of 0.68 pg/mg, LOQ of 2.4 pg/mg, and precision below 8.12%. Since EtG was seen to display prolonged stability in the aqueous extracts and after derivatization with PFPA this straightforward procedure allows a routine throughput of large quantities of samples with little proneness to procedural scatter of results. The method was applied to demonstrate the homogeneity of two hair reference materials with mean EtG contents of 8.48 pg/mg and 22.0 pg/mg. Aside from the application in homogeneity studies of hair reference materials predominantly in the concentration range of 10-50 pg/mg the method was also designed for daily routine quantification of real-world sample with regard to drinking behavior assessment.

The detection and quantification of ethyl glucuronide in placental tissue and placental perfusate by headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry.

BACKGROUND: Ethyl glucuronide (EtG) is arising as a promising biomarker of heavy prenatal alcohol exposure, however its transfer across the human placenta is still unclear and is currently being investigated using the ex vivo placental perfusion model. This model allows for sampling from placental tissue and placental perfusate, which is a surrogate to plasma. OBJECTIVE: To develop a method for detecting and quantifying EtG in placental perfusate and tissue using headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). METHODS: A method was optimized by manipulation of the following components to attain the highest peak counts for the quantifying ions of EtG and its deuterated internal standard on the mass spectrum: cartridges used for solid phase extraction, injection method, derivatizing agent, pre-injection parameters, SPME fiber, GC ramp speed, and GC column flow. RESULTS: The final method utilized involved solid phase extraction of standards via UCT CleanScreen Cartridges, derivatization with heptafluorobutyric acid, and introduction into the GC via HS-SPME with adsorption to a polydimethylsiloxane fiber. The method has improved sensitivity over other methods that quantify EtG in blood using GC-MS, with detection limits of 1.6 ng/mL and 13.7 ng/g for placental perfusate and tissue, respectively. The method was applied to samples collected from the fetal reservoir during the ex vivo placental perfusion model and EtG was detected in the fetal circulation after 20 minutes of perfusion, indicating transfer of EtG. CONCLUSIONS: The present method is sensitive and can be used to quantify EtG transfer during ex vivo placental perfusion experiments.