Phosphatidylethanol (PEth) and ethyl glucuronide (EtG) are established biomarkers of alcohol consumption, offering complementary insights into long-term and recent drinking behavior, as well as alcohol tolerance or dependence. This study examined the association between blood alcohol concentration (BAC), EtG, and PEth in blood samples collected predominantly in traffic-related forensic cases. A total of 100 consecutive blood samples from 87 individuals submitted to the Institute of Legal Medicine in Munich for BAC determination were retrospectively analyzed for PEth and EtG. Samples originated from individuals in North Rhine-Westphalia, including mainly driving under influence (DUI), but also other alcohol-related offenses. BAC was measured via gas chromatography with a flame ionization detector and an enzymatic (ADH) method. PEth and EtG were quantified by validated LC-MS/MS procedures. Thirteen samples were excluded as post-drinking pairs from the same individuals, resulting in n = 87 samples included in the analysis. The study population was predominantly male (77

Ethanol is metabolized by alcohol dehydrogenase to acetaldehyde and induces cytochrome P450 2E1 (CYP2E1), which generates reactive oxygen species that cause inflammatory liver damage. Clomethiazole, a drug approved for alcohol withdrawal treatment (AWT) in some European countries, inhibits CYP2E1. We hypothesized that clomethiazole would lead to a faster reduction in oxidative stress, inflammatory cytokines, and liver enzymes compared to diazepam treatment. We analysed respective biomarkers in 50 patients undergoing AWT and 25 healthy individuals but found no statistical difference between the two medication groups over 3–5 days. Hence, our hypothesis was not confirmed during this observation period.

Cannabis is the world's most used illegal drug. The main psychoactive component of cannabis is Δ9-tetrahydrocannabinol (THC). To aid the identification of cannabis-impaired individuals, a simple but effective workflow for reliable quantification of THC and its metabolites in oral fluid samples collected with the Greiner Bio-One Saliva Collection System is presented. Sampling involves rinsing the oral cavity with an extraction solution containing a citrate buffer stimulating salivary flow. Sample processing targeted the cannabinoid fraction interacting with proteins and other insoluble constituents that can be separated by centrifugation. Approximately 50% of the total amount of cannabinoids included in the oral fluid was recovered from the obtained pellet by extraction with acetonitrile. Liquid chromatography-tandem mass spectrometry was used for cannabinoid quantification. Fitness of the developed workflow for application in forensic and clinical cannabis testing was demonstrated by evaluating multiple performance parameters, including selectivity, linearity, limits of quantification (LOQs), accuracy, precision, matrix effects, extraction recoveries, process efficiencies and stability. Furthermore, sensitivity and specificity of the developed oral fluid-based cannabis test was demonstrated by analysing 195 samples collected either from opioid addicts or persons suspected of driving under the influence of drugs. The accuracy of identifying a person with the presence of THC in blood was found to be 97.9%.

Blood sampling is a common practice to monitor health, but it entails a series of drawbacks for patients including pain and discomfort. Thus, there is a demand for more convenient ways to obtain samples. Modern analytical techniques enable monitoring of multiple bioanalytes in smaller samples, opening possibilities for new matrices, and microsampling technologies to be adopted. Interstitial fluid (ISF) is an attractive alternative matrix that shows good correlation with plasma concentration dynamics for several analytes and can be sampled in a minimally invasive and painless manner from the skin at the point-of-care. However, there is currently a lack of sampling devices compatible with clinical translation. Here, to tackle state-of-the-art limitations, a cost-effective and compact single-microneedle-based device designed to painlessly collect precisely 1.1 µL of dermal ISF within minutes is presented. The fluid is volume-metered, dried, and stably stored into analytical-grade paper within the microfluidic device. The obtained sample can be mailed to a laboratory, quantitatively analyzed, and provide molecular insights comparable to blood testing. In a human study, the possibility to monitor various classes of molecular analytes is demonstrated in ISF microsamples, including caffeine, hundreds of proteins, and SARS-CoV-2 antibodies, some being detected in ISF for the first time.

This registered clinical trial sought to validate a laboratory test system devised to screen medications for alcoholism treatment (TESMA) under different contingencies of alcohol reinforcement. Forty-six nondependent, but at least medium-risk drinkers were given the opportunity to earn intravenous infusions of ethanol, or saline, as rewards for work in a progressive-ratio paradigm. Work demand pattern and alcohol exposure dynamics were devised to achieve a gradual shift from low-demand work for alcohol (WFA) permitting quickly increasing breath alcohol concentrations (BrAC) to high-demand WFA, which could only decelerate an inevitable decrease of the previously earned BrAC. Thereby, the reward contingency changed, modeling different drinking motivations. The experiment was repeated after at least 7 days of randomized, double-blinded treatment with naltrexone, escalated to 50 mg/d, or placebo. Subjects treated with naltrexone reduced their cumulative WFA (cWFA) slightly more than participants receiving placebo. This difference was not statistically significant in the preplanned analysis of the entire 150 min of self-administration, i.e., our primary endpoint ( p = 0.471, Cohen’s d = 0.215). Naltrexone serum levels correlated with change in cWFA ( r = −0.53; p = 0.014). Separate exploratory analyses revealed that naltrexone significantly reduced WFA during the first, but not the second half of the experiment (Cohen’s d = 0.643 and 0.14, respectively). Phase-dependent associations of WFA with changes in subjective stimulation, wellbeing and desire for alcohol suggested that the predominant reinforcement of WFA was positive during the first phase only, and might have been negative during the second. We conclude that the TESMA is a safe and practical method. It bears the potential to quickly and efficiently screen new drugs for their efficacy to attenuate positively reinforced alcohol consumption. It possibly also provides a condition of negative reinforcement, and for the first time provides experimental evidence suggesting that naltrexone’s effect might depend on reward contingency.

BACKGROUND:Therapeutic drug monitoring (TDM) is recommended for opioid maintenance therapy with levomethadone. However, TDM has not yet been applied to monitor opioid withdrawal therapy clinically, although tools to improve it are required.METHODS:In this observational cohort study, repeated TDM with levomethadone was performed according to a prospective opioid withdrawal study protocol. Objective and subjective opioid withdrawal symptoms were measured using validated rating scales and correlated to levomethadone plasma concentrations. Plasma levels were measured using high-pressure liquid chromatography with column switching and spectroscopic detection of methadone and its major metabolite.RESULTS:This study included 31 opioid-dependent patients who participated in standardized opioid withdrawal therapy. The serum levels of levomethadone were found to be highly variable and below the recommended therapeutic reference range of 250 ng/mL for maintenance therapy. These serum levels were positively correlated with dosage (r = 0.632; P < 0.001) and inversely correlated with subjective (r = -0.29; P = 0.011) and objective (r = -0.28; P = 0.014) withdrawal symptoms.CONCLUSIONS:The evidence provided sheds light on how to improve levomethadone withdrawal therapy in patients with opioid dependence. It seems likely that higher initial doses at the beginning and lower dose reductions would have been advantageous. TDM can enhance the safety of opioid withdrawal therapies, minimize withdrawal symptoms, and reduce dropout rates.

The prevalence of drug use among nightlife attendees needs to be accurately estimated to, for example, evaluate preventive interventions. This study tested the feasibility of using a breath-sampling device to estimate the prevalence of drug use among nightlife attendees. The study was conducted at five nightclubs and a large music festival in Stockholm, Sweden. Participants were invited to participate and microparticles in exhaled breath were sampled and analyzed for 47 compounds using a state-of-the-art analytic method that follows forensic standards. In addition, participants’ breath alcohol concentration was measured and they were interviewed about demographics, drinking habits, and drug use. Of the people invited, 73.7% (n = 1223) agreed to participate, and breath samples were collected from 1204 participants. Breath sampling was fast and well-accepted by participants. 13 percent of participants tested positive for an illicit drug, but only 4.3% self-reported drug use during the last 48 h. The most common substances detected were cocaine, amphetamine, and MDMA. There was no agreement between self-reported and measured use of any drug. Breath sampling is a convenient method to test illicit drug use among a large number of participants at events, and can be used as an estimate of drug use prevalence.

Little is known about the impact of morphological disorders in distinct zones on metabolic zonation. It was described recently that periportal fibrosis did affect the expression of CYP proteins, a set of pericentrally located drug-metabolizing enzymes. Here, we investigated whether periportal steatosis might have a similar effect. Periportal steatosis was induced in C57BL6/J mice by feeding a high-fat diet with low methionine/choline content for either two or four weeks. Steatosis severity was quantified using image analysis. Triglycerides and CYP activity were quantified in photometric or fluorometric assay. The distribution of CYP3A4, CYP1A2, CYP2D6, and CYP2E1 was visualized by immunohistochemistry. Pharmacokinetic parameters of test drugs were determined after injecting a drug cocktail (caffeine, codeine, and midazolam). The dietary model resulted in moderate to severe mixed steatosis confined to periportal and midzonal areas. Periportal steatosis did not affect the zonal distribution of CYP expression but the activity of selected CYPs was associated with steatosis severity. Caffeine elimination was accelerated by microvesicular steatosis, whereas midazolam elimination was delayed in macrovesicular steatosis. In summary, periportal steatosis affected parameters of pericentrally located drug metabolism. This observation calls for further investigations of the highly complex interrelationship between steatosis and drug metabolism and underlying signaling mechanisms.

Drug Testing and AnalysisVolume 15, Issue 7 p. 791-792 LETTER TO THE EDITOR Comment on the upper cutoff level for the alcohol biomarker phosphatidylethanol (PEth) for the assessment of alcohol consumption in forensic practice Frank Musshoff, Corresponding Author Frank Musshoff [email protected] orcid.org/0000-0002-9613-0525 Forensisch Toxikologisches Centrum GmbH, Munich, Germany Correspondence Frank Musshoff, Forensisch Toxikologisches Centrum GmbH, Dessauerstr. 13-15, 80992 Munich, Germany. Email: [email protected]Search for more papers by this authorMichael Böttcher, Michael Böttcher MVZ Labor Dessau GmbH, Dessau-Roßlau, GermanySearch for more papers by this authorMatthias Graw, Matthias Graw Institute of Legal Medicine, Ludwig-Maximilians-University of Munich, Munich, GermanySearch for more papers by this authorGisela Skopp, Gisela Skopp Forensisch Toxikologisches Centrum GmbH, Munich, GermanySearch for more papers by this authorJasna Neumann, Jasna Neumann MVZ Labor Dessau GmbH, Dessau-Roßlau, GermanySearch for more papers by this authorGudrun Høiseth, Gudrun Høiseth Department of Forensic Sciences, Oslo University Hospital, Oslo, NorwaySearch for more papers by this authorAnders Helander, Anders Helander orcid.org/0000-0001-7480-8078 Department of Laboratory Medicine, C1:74, Karolinska University Hospital, Karolinska Institutet, Stockholm, SwedenSearch for more papers by this author Frank Musshoff, Corresponding Author Frank Musshoff [email protected] orcid.org/0000-0002-9613-0525 Forensisch Toxikologisches Centrum GmbH, Munich, Germany Correspondence Frank Musshoff, Forensisch Toxikologisches Centrum GmbH, Dessauerstr. 13-15, 80992 Munich, Germany. Email: [email protected]Search for more papers by this authorMichael Böttcher, Michael Böttcher MVZ Labor Dessau GmbH, Dessau-Roßlau, GermanySearch for more papers by this authorMatthias Graw, Matthias Graw Institute of Legal Medicine, Ludwig-Maximilians-University of Munich, Munich, GermanySearch for more papers by this authorGisela Skopp, Gisela Skopp Forensisch Toxikologisches Centrum GmbH, Munich, GermanySearch for more papers by this authorJasna Neumann, Jasna Neumann MVZ Labor Dessau GmbH, Dessau-Roßlau, GermanySearch for more papers by this authorGudrun Høiseth, Gudrun Høiseth Department of Forensic Sciences, Oslo University Hospital, Oslo, NorwaySearch for more papers by this authorAnders Helander, Anders Helander orcid.org/0000-0001-7480-8078 Department of Laboratory Medicine, C1:74, Karolinska University Hospital, Karolinska Institutet, Stockholm, SwedenSearch for more papers by this author First published: 30 December 2022 https://doi.org/10.1002/dta.3432Citations: 2Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. REFERENCES 1Luginbühl M, Wurst FM, Stöth F, Weinmann W, Stove CP, van Uytfanghe K. Consensus for the use of the alcohol biomarker phosphatidylethanol (PEth) for the assessment of abstinence and alcohol consumption in clinical and forensic practice (2022 Consensus Basel). Drug Testing Anal. 2022; 14(10): 1800-1802. doi:10.1002/dta.3340 2Ulwelling W, Smith K. The PEth blood test in the security environment: what it is; why it is important; and interpretative guidelines. J Forensic Sci. 2018; 63(6): 1634-1640. doi:10.1111/1556-4029.13874 3Helander A, Hansson T. Nationell harmonisering av alkoholmarkören PEth [National harmonization of the alcohol biomarker PEth]. Lakartidningen. 2013; 110(39–40): 1747-1748. PMID: 24245431. 4 Deutsche Gesellschaft für Verkehrspsychologie (DGVP) und Deutsche Gesellschaft für Verkehrsmedizin (DGVM). Urteilsbildung in der Fahreignungsbegutachtung - Beurteilungskriterien, 4. Bonn, Germany: Auflage, Kirschbaum Verlag; 2022. 5Schröck A, Pfäffli M, König S, Weinmann W. Application of phosphatidylethanol (PEth) in whole blood in comparison to ethyl glucuronide in hair (hEtG) in driving aptitude assessment (DAA). Int J Leg Med. 2016; 130(6): 1527-1533. doi:10.1007/s00414-016-1394-4 6Weinmann W, Schröck A, Wurst FM. Commentary on the paper of Walther L. et al.: phosphatidylethanol is superior to CDT and GGT as an alcohol marker and is a reliable estimate of alcohol consumption level. Alcohol Clin Exp Res. 2016; 40(2): 260-262. doi:10.1111/acer.12946 7Helander A, Hermansson U, Beck O. Dose–response characteristics of the alcohol biomarker phosphatidylethanol (PEth)—a study of outpatients in treatment for reduced drinking. Alcohol Alcohol. 2019; 54(6): 567-573. doi:10.1093/alcalc/agz064 8Isaksson A, Walther L, Hansson T, Andersson A, Stenton J, Blomgren A. High-throughput LC-MS/MS method for determination of the alcohol use biomarker phosphatidylethanol in clinical samples by use of a simple automated extraction procedure-preanalytical and analytical conditions. J Appl Lab Med. 2018; 2(6): 880-892. doi:10.1373/jalm.2017.024828 9Årving A, Høiseth G, Hilberg T, et al. Comparison of the diagnostic value of phosphatidylethanol and carbohydrate-deficient transferrin as biomarkers of alcohol consumption. Alcohol Clin Exp Res. 2021; 45(1): 153-162. doi:10.1111/acer.14503 10Neumann J, Beck O, Böttcher M. Phosphatidylethanol, ethyl glucuronide and ethanol in blood as complementary biomarkers for alcohol consumption. J Mass Spectrom Adv Clin Lab. 2021; 22: 3-7. doi:10.1016/j.jmsacl.2021.09.005 11Høiseth G, Hilberg T, Trydal T, Husa A, Vindenes V, Bogstrand ST. The alcohol marker phosphatidylethanol is closely related to AST, GGT, ferritin and HDL-C. Basic Clin Pharmacol Toxicol. 2022; 130(1): 182-190. doi:10.1111/bcpt 12Helander A, Böttcher M, Dahmen N, Beck O. Elimination characteristics of the alcohol biomarker phosphatidylethanol (PEth) in blood during alcohol detoxification. Alcohol Alcohol. 2019; 54(3): 251-257. doi:10.1093/alcalc/agz027 13Gnann H, Weinmann W, Thierauf A. Formation of phosphatidylethanol and its subsequent elimination during an extensive drinking experiment over 5 days. Alcohol Clin Exp Res. 2012; 36(9): 1507-1511. doi:10.1111/j.1530-0277.2012.01768.x Citing Literature Volume15, Issue7July 2023Pages 791-792 ReferencesRelatedInformation

Perinatal illicit drug consumption has become a critical concern because of associated adverse mental and physical outcomes in the child. Drug screening in meconium can identify prenatal drug exposure beginning at the 12th week of gestation and therefore is gaining interest in pediatric care. In our laboratory, meconium is routinely analysed with an EN15189 accredited UPLC-MS/MS Multi-Target-Screening method (MTS) on a Waters Acquity UPLC connected to a Xevo TQ-XS. The method targets a panel of 65 licit and illicit drugs at LLOQs between 0.1 and 2 ng/g meconium. In addition, a commercial library based "known-unknown" screening is performed with the Bruker ToxTyper™ (TT) system. In this study, we describe our analytical findings from routine to compare the performance of these two methods. A 200 μL aliquot of a homogenized meconium suspension (1 mL H2O per 100 mg meconium) is fortified with deuterated internal standards for MTS (n = 65) or TT (n = 12) and subsequently hydrolysed with β-glucuronidase B-ONE (Kura Biotech) for 30 min at room temperature. Thereafter, a salting out assisted-liquid/liquid-extraction is conducted by adding 800 μL 10 M ammonium acetate/acetonitrile (1/3, v/v). After centrifugation 300 μL from the organic supernatant is transferred into glass vials and evaporated into 15 μL ethylene glycol under N2 at 45 °C. The residue is then diluted in mobile phase A and measured by MTS and TT method applying a library search in the Bruker (980 entries) and Maurer/Wissenbach/Weber library (2nd edition 2019, > 4000 entries). The majority of the samples (58%) came from the German states of Bavaria, Saxony and Thuringia. From 479 consecutive routine samples analysed, 270 (56%) contained illicit substances and 145 (30%) contained prescription/OTC drugs. The remaining 64 (14%) samples presented negative in both methods. With MTS, 41 out of 65 possible analytes were found, among which amphetamine (n = 124, 26%) and methamphetamine (n = 101, 21%) revealed a high positive rate. With TT however, only 79 (16%) and 84 (18%) samples were positive, resulting in an agreement of 64% and 83%, respectively. THC and its metabolites OH-THC and THC-COOH showed different detection rates with MTS and TT; 119 and 2 THC positive samples (agreement 1.7%), 152 and 85 11-OH-THC positive samples (agreement 56%) and 144 and 43 THC-COOH positive samples (agreement 30%). The positive rates of MTS for other abuse relevant drugs were: morphine 5.8%, codeine 4.4%, ketamine 4.8% and cocaine 1.5%. Agreement with TT was: 57% for morphine, 91% for codeine, 61% for ketamine and 43% for cocaine. TT never found a substance from the MTS panel alone. Based on the comprehensive databases, additional 52 drugs could be reported by TT. In 16 cases, pethidine and piritramide were detected; all other substances were prescription drugs in some cases related to pregnancy. The concentrations of most parent drugs observed in meconium were generally lower than their main metabolites, such as methadone, buprenorphine, cannabis, cocaine, ketamine. Our work revealed that TT had a lower detection rate than MTS with an agreement of < 80% for most of the detected parameters. Furthermore, the majority of the additional drugs identified by the TT approach were of no clinical relevance. Cannabis, methamphetamine and amphetamine had the highest prevalence, which can be rated as a southeast Germany phenomenon. Interestingly, the cannabis metabolite OH-THC had the highest positive rate (32%) and the highest concentration of the 3 cannabinoids. The positive rates for drugs of abuse underline the usefulness of meconium for drug testing. The best future strategy would be to extend the MTS method rather than continue with the cost-intensive TT method.

Comparing the sensitivity of the Waters UniSpray™ ion source (USI) with the Waters electrospray ion source (ESI) on a Waters Xevo TQ-XS mass spectrometer with and without additional post-column infusion of low concentrated hydrochloric acid (HCl). To reduce sample volumes or increase analytical sensitivity for drugs of abuse detection in human body fluids, technological advances in mass spectrometry often target an increase in ionization efficiency. One example for this development is the USI, which claims higher ion yields and therefore higher sensitivity due to a unique geometry. Another possibility to increase sensitivity through increased ion yield is the post-column infusion of low concentrated HCl. For sensitivity comparison, we choose the, in our opinion, 119 most relevant medications and drugs of abuse. These were prepared in eleven acetonitrile solutions containing 10 analytes each and one with 9 analytes at a concentration of 1 μg/mL for each substance. For analysis, every solution was diluted 1:1000 in mobile phase A/ethylene glycol (70:30 v/v) to a concentration of 1.0 ng/mL. Solutions were analysed in triplicate on the same day interrupted by the ion source change, to decrease inter-day variations in sensitivity. Chromatographic separation was performed on a Waters 2.1 × 150 mm, 1.8 μm BEH Phenyl column kept at 60 °C on an Acquity UPLC connected to a Xevo TQ-XS detector (Waters). The standard ESI ion source or the USI ion source was mounted on the detector. Mobile phase A consisted of 20 mM ammonium formate and 0.1% formic acid (pH 3) and mobile phase B was 0.1% formic acid in methanol. Gradient elution was conducted within 6.0 min at a flow rate of 0.5 mL/min with 85% A and 15% B at the beginning and 100% B at the end. Data were acquired with the ion sources operating in the positive ionization MRM mode. Two transitions for each analyte were monitored at a constant dwell time of 10 ms over the full chromatographic run time. In an additional experiment, both ion sources were compared with additional post-column infusion of 0.0037% HCl at a flow rate of 5.0 μL/min. Injection volume was 2.0 μL in all experiments. USI provided a general gain in signal intensity (peak area): 12% (n = 14) of the analytes showed a peak area increase up to 2-fold, 85% (n = 101) increased between 2- and 4-fold and 3% (n = 4) increased above 4-fold in peak area. Post-column HCl infusion with the ESI source mounted, resulted in an analyte dependent additional gain in signal intensity for 15% (n = 18) of the analytes ranging from 2- to 21-fold. 78% (n = 93) ranged from 0.8-fold to 2-fold. In contrast 7% (n = 8) showed a signal decrease between 0.4-fold and 0.8-fold. The combination of USI and post-column HCl infusion resulted in an increase in peak area from 2-fold up to 62-fold compared to ESI without infusion for 91% (n = 108) of the analytes. 8% (n = 9) where between 0.8-fold and 2-fold and 1% (n = 2) ranged from 0.4-fold to 0.8-fold. With USI, a general improvement in analytical sensitivity under identical conditions could be achieved for all of our 119 "favorite analytes" separated in one chromatographic run. Additional post-column infusion of HCl increased sensitivity for many substances. This approach can be used very selectively when applied to certain retention time windows only. Further experiments are needed to understand how different adjustments of ESI and USI and different matrices will affect ionization. USI, especially in combination with post-column HCl infusion, is a promising tool to increase sensitivity in toxicological multi-target LC-MS/MS analysis.

Background: Vitreous humor (VH) is a specimen of great value in forensic investigations and is being used for evaluating possible post-mortem formation of ethanol. Ethyl glucuronide (EtG) is an ethanol metabolite that has found interest for the same purpose. Both compounds can be measured in VH and because of differences in rate of distribution and elimination they may offer complementary information. Methods: VH, femoral blood (FB) and urine were collected from 117 autopsy cases for forensic investigation. Ethanol was measured with headspace gas chromatography, while EtG was measured with liquid chromatography - tandem mass spectrometry. Results and conclusion: Ethanol was detected in all matrices in 39 cases, while EtG was present in 62 cases. The VH-FB and the VH-urine ethanol concentrations in the 39 cases were statistically correlated (p < 0.00001). In one case with an ethanol concentration of 0.11 g/L in FB, no ethanol was detected in VH and urine, and no EtG in any specimen, indicating a possible post-mortem formation. EtG was present in VH in more cases than in FB and urine. The correlation between the EtG concentrations in VH and FB was statistically significant (p < 0.0003) as was the case also for VH and urine (p < 0.001). The combined information on ethanol and EtG concentrations in the three matrices can be used to interpret alcohol drinking habit before death. This study confirms the value of using VH as a specimen in forensic investigations regarding recent exposure to ethanol. EtG can be used not only for investigating postmortem ethanol formation but also for estimating recent alcohol drinking. (C) 2020 Published by Elsevier B.V.

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

This study aimed to determine whether hydromorphone and codeine can be detected in oral fluid specimens following administration of Substitol™, a slow-release formulation of morphine. This is of interest for those monitoring treatment compliance using drug testing. Oral fluid specimens collected for compliance assessment in routine clinical practice or as part of a clinical trial were subjected to quantitative analysis of hydromorphone, morphine, codeine, and 6-acetylmorphine using highly sensitive mass spectrometric methods. Oral fluid was collected using a Greiner Bio-One saliva collection system. Patients undergoing substitution treatment with Substitol™, methadone, or buprenorphine were included, together with patients undergoing pain treatment with hydromorphone. Hydromorphone was detected in 642 of the 663 (97%) samples from substitol-treated patients. Concentrations were not higher in methadone- and buprenorphine-treated patients who relapsed into heroin use, or in patients on hydromorphone therapy. Codeine was detected in 29% of the samples. These concentrations were lower than those in patients who had relapsed to heroin use. Clinical administration of morphine can lead to detectable concentrations of both hydromorphone and codeine in oral fluids. This should be taken into consideration when using drug testing in oral fluid samples for compliance assessment in this patient group.

Purpose/Background Ketamine (K) is used as a party drug with hallucinogenic properties with a half-life of about 2.5 hours. Data are available with respect to the detection window (ie, when a person is still tested positive for the drug and/or metabolite after use) of K after single use. Nevertheless, no data are available with respect to the detection window of K in urine after chronic use. Methods/Procedures This retrospective case series describes 7 patients with K dependency as their main addiction who have been admitted to an addiction center for K detoxification. Their abstinence-oriented care involved routine urinary screening of K and its metabolites, as well as traditional drugs of abuse, such as cocaine and cannabinoids. Findings/Results Urine samples remained positive for all the cases identified after 22 to 96 days. A peak detection period of 61, 40, and 96 days for K, norketamine, and dehydronorketamine, respectively, measured using liquid chromatography-tandem mass spectrometry at a cutoff concentration of 1.0 ng/mL, is defined. The K/norketamine and K/dehydronorketamine ratios varied over time between 0.33 and 3.06, and 0.01 and 0.36 for all patients, respectively, implying a large interindividual variation in K metabolism. Implications/Conclusions Ketamine and its metabolites have a prolonged excretion profile in urine, which requires frequent measurements (at least weekly) to guide abstinence treatment. Further research is needed to develop an algorithm that can differentiate new K use from residual urinary K excretion in urine of chronic daily users.

Abstract Aims To compare the performance of short- and long-term alcohol biomarkers for the evaluation of alcohol drinking in employment-related health controls. Methods The 519 blood samples originated from 509 patients (80% men) presenting at occupational health units and medical centers at employment agencies for the evaluation of risky drinking. The laboratory investigation comprised the measurement of phosphatidylethanol (PEth 16:0/18:1), carbohydrate-deficient transferrin (CDT; % disialotransferrin), gamma-glutamyl transferase (GGT), mean corpuscular volume (MCV), ethanol and ethyl glucuronide (EtG). Results Many samples tested positive for acute (57%) and chronic (69%) alcohol biomarkers. PEth was the single most positive biomarker (64%; cut-off 0.05 μmol/l or 35 μg/l) and the only positive chronic biomarker in 100 cases. The highest PEth concentrations were seen in samples positive for all chronic biomarkers, followed by those also being CDT positive (cut-off 2.0%). All 126 CDT-positive samples were positive for PEth using the lower reporting limit (≥0.05 μmol/l) and for 114 cases (90%) also using the higher limit (≥0.30 μmol/l or 210 μg/l). In the CDT-positive cases, the PEth median concentration was 1.71 μmol/l, compared with 0.45 μmol/l for the CDT-negative cases (P < 0.0001). PEth and CDT values were correlated significantly (r = 0.63, P < 0.0001). Among the EtG-positive cases (≥1.0 ng/ml), 95% were also PEth positive, and all ethanol-positive cases (≥0.10 g/l) were also PEth positive. Conclusions For optimal detection of drinking habits, using a combination of short- and long-term alcohol biomarkers provided best information. PEth was the single most positive alcohol biomarker, whereas GGT and MCV offered little additional value over PEth and CDT.

Buprenorphine is a partial μ-opioid agonist widely used for opioid maintenance therapy (OMT). It is mainly metabolized to pharmacologically active norbuprenorphine by the cytochrome P450 (CYP) isozyme 3A4. This may give rise to drug–drug interactions under combinations with inhibitors or inducers of CYP3A4. Cannabis is a potential inhibitor of CYP3A4, and there is a large degree of concomitant cannabis use among OMT patients. We performed a retrospective analysis on liver healthy OMT patients substituted with buprenorphine, either with (n = 15) or without (n = 17) concomitant use of cannabis. Patients with additional illicit drugs or medications affecting CYP3A were excluded. Measured blood concentrations of buprenorphine and norbuprenorphine were compared between the two groups. Cannabis users and non-users received similar doses, but users had 2.7-fold higher concentrations of buprenorphine (p < 0.01) and 1.4-fold for norbuprenorphine (1.4-fold, p = 0.07). Moreover, the metabolite-to-parent drug ratio was 0.98 in non-users and 0.38 in users (p = 0.02). Female gender did not produce significant effects. These findings indicate that cannabis use decreases the formation of norbuprenorphine and elevates buprenorphine and norbuprenorphine concentrations in blood most probably by inhibition of CYP3A4. The pharmacokinetic interaction may give rise to enhanced or altered opioid activity and risk of intoxications. Physicians should inform patients about this risk and supervise cannabis users by regular control of buprenorphine blood levels, i.e., by therapeutic drug monitoring.

Abstract Objectives Ethyl glucuronide (EtG) is a conjugated, minor ethanol metabolite used as a biomarker for recent alcohol intake. EtG is commonly measured in urine as part of a drug testing service but has also attracted attention for measurement in blood. However, due to lower EtG concentrations in blood, the detection time is expected to be shorter. The present work aimed to improve the analytical sensitivity of EtG in blood, to prolong the detection time. Methods A liquid chromatography-tandem mass spectrometry method was developed for EtG in whole blood and serum, using protein precipitation with methanol, a deuterated internal standard, and selected reaction monitoring mode with negative electrospray ionization. No significant matrix effect was observed. The method generated linear results in the measuring range 1.0–50 μg/L, the accuracy was within ±10% and the imprecision <15%. Results In 46 patients followed with daily blood and urine sampling during alcohol detoxification, the mean (median) time to first negative serum EtG sample was 112 (111) h. This was slightly longer than for EtG in urine, using 100 μg/L as cutoff. The detection rate was 76% for serum EtG and 68% for urine EtG. In cases where serum EtG was positive but urine EtG negative, the urine samples tended to be more dilute as indicated by lower creatinine concentrations. On admission to hospital, the whole-blood and serum EtG concentrations correlated with the breath ethanol concentration (p = 0.012 and p = 0.027, respectively). In 100 patients sampled at admission to hospital for other reasons than substance abuse and with no ethanol detected in breath, 40% tested positive for EtG in serum and 43% in urine. In 79 paired urine and serum EtG measurements, the median urine/serum concentration ratio was 155. Conclusions A sensitive method was developed for EtG measurement in whole-blood and serum specimens, offering similar detection time for recent alcohol exposure compared with routine EtG measurement in urine.

BACKGROUND:Detection of heroin use is among the major tasks for drug testing and can be best performed by using 6-acetylmorphine as the target analyte. This study was performed to document analytical findings in oral fluid after OF heroin intake. METHODS:The samples were from routine drug testing of patients in substitution treatment. The analytical investigation was made with a forensic accredited liquid chromatography-tandem mass spectrometry method. RESULTS:Out of 2814 samples, from 1875 patients, sent for routine drug testing, 406 contained one or more opiate in the drug screening when applying a cutoff limit of 1 ng/mL neat OF. Out of these 406, 314 had a measured 6-AM concentration in neat OF ≥ 1 ng/mL. The study demonstrated that 6-AM is a viable parameter in oral fluid drug testing with an about 80% sensitivity compared to using morphine and codeine as biomarkers. An additional value of using 6-AM is the confidence in concluding a heroin intake. The 6-AM concentrations varied between 1 and >1000 ng/mL, with a median value of 18.6 ng/mL. Heroin was measured in 35 samples with a median value of 0.72 ng/mL. The positive rate for opiates in urine and OF drug testing was the same, 13.5%, in similar populations of patients. CONCLUSIONS:6-AM is a preferred parameter in OF drug testing for monitoring heroin use and makes OF drug testing for detecting heroin use more effective than urine drug testing when using highly sensitive mass spectrometry methods.