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The repeatability and extraction recoveries of parallel electromembrane extraction (Pa-EME) was thoroughly investigated in the present project. Amitriptyline, fluoxetine, and haloperidol were isolated from eight samples of pure water, undiluted human plasma, and undiluted human urine, respectively; in total 24 samples were processed in parallel. The repeatability was found to be independent of the different sample matrices (pure water samples, human plasma, and water) processed in parallel, although the respective samples contained different matrix components. In another experiment seven of the 24 wells were perforated. Even though the perforation caused the total current level in the Pa-EME setup to increase, the intact circuits were unaffected by the collapse in seven of the circuits. In another approach, exhaustive extraction of amitriptyline, fluoxetine, and haloperidol was demonstrated from pure water samples. Amitriptyline and haloperidol were also isolated exhaustively from undiluted human plasma samples; the extraction recovery of fluoxetine from undiluted human plasma was 81%. Finally, the sample throughput was increased with the Pa-EME configuration. The extraction recoveries were investigated by processing 1, 8, 68, or 96 samples in parallel in 10min; neither the extraction recoveries nor the repeatability was affected by the total numbers of samples. Eventually, the Pa-EME was combined with ultra performance liquid chromatography (UPLC) to combine high-throughput sample preparation with high-throughput analytical instrumentation. The aim of the present investigation was to demonstrate the potential of electromembrane extraction as a high throughput sample preparation platform; and hopefully to increase the interest for EME in the bioanalytical field to solve exisiting and novel analytical challenges., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: A large proportion of road traffic crashes are related to driving under the influence (DUI) of alcohol or drugs. The aim of this study was to compare the use of alcohol, illegal drugs and psychoactive medicinal drugs among random drivers in Brazil and Norway, two countries with the same legal limit for drunk driving, but with marked differences in legislation history, enforcement and penalties for DUI, and to discuss any differences found., Methods: Roadside surveys were conducted on Fridays and Saturdays between noon and midnight. Samples of oral fluid were collected for analysis of drugs, whereas alcohol was determined by breath testing or by analysis of oral fluid., Results: High participation rates of 94-97% were obtained in both countries. The weighted prevalence of driving with alcohol concentrations in breath or oral fluid equivalent to blood alcohol concentrations (BAC) above 0.2g/L was 2.7% (95% CI 2.2-3.3) in Brazil and 0.2% (95% CI 0.0-0.5) in Norway. Stimulants (amphetamines or cocaine) were found in samples from 1.0% (95% CI 0.7-1.4) of drivers in Brazil and 0.3% (95% CI 0.1-0.7) in Norway. The prevalence of amphetamines was highest among Brazilian truck drivers (3.6%; 95% CI 2.0-6.4). Tetrahydrocannabinol was found in samples from 0.5% (95% CI 0.3-0.8) of drivers in Brazil and 1.0% (95% CI 0.6-1.5) in Norway, whereas benzodiazepines or zopiclone were found in 1.0% (95% CI 0.7-1.4) and 1.7% (95% CI 1.2-2.4) of the samples from Brazil and Norway, respectively., Conclusions: The difference in the prevalence of alcohol may be related to the fact that Norway has implemented steps to reduce drunk driving since 1936, whereas Brazil has attempted to do the same for only a few years. Differences for drugs may be related to different patterns in the use of stimulants, cannabis and medicines., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Working under the influence of drugs and/or alcohol may affect safety and job performance. However, the size of this possible problem among health professionals (HPs) is unknown. The aim of this study was threefold: (i) to analyze samples of oral fluid and self-reported data from questionnaires to investigate the prevalence of alcohol and drugs among a sample of HPs in Norway, (ii) to study self-reported absence from or impairment at work due to alcohol and/or drug use, and (iii) to examine whether such use and absence/impairment due to such use depend on socio-demographic variables.A total of 916 of the 933 invited HPs from hospitals and pharmacies participated in the study (participation rate = 98.2%), and 81.1% were women. Associations were analyzed in bi-variate cross tables with Chi-square statistics to assess statistical significance.Alcohol was not detected in any of the samples. Ethyl glucuronide, a specific alcohol metabolite, was found in 0.3% of the collected samples. Illicit drugs and medicinal drugs were identified in 0.6% and 7.3% of the samples, respectively. Both analytical results and self-reported use of alcohol and drugs during the past 12 months indicate that recent and past year alcohol and drug use was lower among HPs than among workers in other business areas in Norway, Europe and US. Nevertheless, several HPs reported absence from work due to alcohol (0.9%) and medicinal drug use (0.8%) during the past 12 months. A substantial part (16.7%) of the self-reported medicinal drug users reported absence from work because of use of medicinal drugs during the past 12 months, and more than 1/4 of those reported in-efficiency at work because of the use of medicinal drugs during the past 12 months. Reduced efficiency at work due to alcohol use during the past 12 months was reported by 12.2%.This sample of HPs seldom used illicit drugs, few had a high level of alcohol consumption, and few tested positive for medicinal drugs. Absence or hangover related to the use of medicinal drugs or alcohol appeared to be a bigger issue than the acute intoxication or the use of illicit drugs.

Background: The abuse of a rapidly changing range of synthetic cannabinoids is increasing worldwide. Oral fluid, which contains the parent compounds and is easily collected, could be a good alternative medium for drug screening for synthetic cannabinoids., Results: A method for screening of 18 synthetic cannabinoids in preserved oral fluid collected with the Intercept® collection device, using UPLC-MS/MS, was validated. Limits of quantification ranged from 0.2 to 2 ng/ml in oral fluid. In several real cases, AM-2201 and/or JWH-018 were found., Conclusion: The presented method allowed rapid and sensitive screening of synthetic cannabinoids in preserved oral fluid collected with the Intercept collection device.

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.

Objective: To examine alcohol and drug use among random drivers in different regions of Norway by analyzing oral fluid, compare drivers in urban and rural areas, compare with results from the roadside survey in southeastern Norway in 2005-2006, and roughly estimate the prevalence of driving with blood drug concentrations above the new Norwegian legislative limits among random drivers. This roadside survey was part of the European DRUID (Driving Under the Influence of Drugs, Alcohol and Medicines) Project., Methods: Drivers were selected for a voluntary and anonymous study using a stratified multistage cluster sampling procedure in collaboration with the Mobile Police Service. Samples of oral fluid were taken using the Statsure Saliva Sample (Statsure Diagnostic Systems, Framingham, MA), and the drivers' gender, age, and nationality were recorded. Samples of oral fluid were analyzed for alcohol or drugs, for a total 28 psychoactive substances., Results: One hundred eighty-four roadside survey sessions were conducted and 10,004 drivers were asked to participate. The refusal rate was 5.8 percent. Psychoactive substances were found in 4.8 percent of the 9410 oral fluid samples analyzed. Alcohol was detected in 0.3 percent, medicinal drugs in 3.2 percent, and illegal drugs in 1.5 percent of the samples. Illegal drugs were significantly more frequently detected in samples from southeastern Norway including the capital Oslo, whereas medicinal drugs were more frequently detected in samples from southeastern Norway excluding Oslo. Illegal drugs were significantly more frequently detected in samples from drivers in urban areas than in rural areas, though there were no significant differences for alcohol and medicinal drugs. Medicinal drugs were most commonly found in samples collected during the daytime on weekdays (3.8%), and illegal drugs were most commonly found in samples collected during late night on weekdays or weekends (2.8%-3.2%). The most commonly found substances were the sleeping agent zopiclone (1.4%), the main active substance in cannabis tetrahydrocannabinol (1.1%), and the sedative drug diazepam (0.7%). The prevalence of driving with drug concentrations above the Norwegian legislative limits for blood was estimated to be about 0.2 percent for alcohol, 0.6 percent for illegal drugs, and about 1.3 percent for medicinal drugs., Conclusions: The incidence of drink driving was very low, though driving after using psychoactive illegal or medicinal drugs was more frequent.

Introduction: The pharmacokinetic profile and systemic bioavailability of a substance is often described by blood or total tissue concentrations. For centrally acting drugs, like opioids, the free fraction of active compound in brain extracellular fluid (ECF) is more likely to be correlated to the pharmacodynamic effects than the blood concentrations. Drugs of abuse, like heroin, are often administered intravenously as bolus injections, and the blood concentrations might change rapidly due to metabolism and distribution. The aim of our study was to establish a method to measure the free fraction of heroin and its metabolites in brain ECF, and follow their fast changes in concentration., Methods: Sprague-Dawley rats were injected intravenously with a bolus of heroin. Heroin and its main metabolites 6-monoacetylmorphine, morphine and morphine-3-glucuronide were measured simultaneously. Brain microdialysis was used for sampling and a method for quantification using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed. Deuterated analogues for each analyte were included in the microdialysis perfusion solution as calibrators for recovery estimation., Results: A highly sensitive UPLC-MS/MS method allowed short sampling intervals, down to one minute, and the simultaneous detection of each analyte and its specific deuterated analogues, making possible the individual recovery calculation for each compound of interest. This method allowed us to determine the pharmacokinetic profiles of heroin and its metabolites in brain-ECF in the same animal after an intravenous injection of heroin., Discussion: Our method makes detecting concurrently the rapid changes in concentrations of heroin and its metabolites in brain ECF possible, despite the rapid metabolism of heroin. Recovery was measured specifically for each analyte in the same sample by carefully combining different deuterated analogues. This technique can be applied to pharmacokinetic studies where more than one compound of interest has to be monitored, and to study distribution of prodrugs or drugs with active metabolites., (Copyright © 2012 Elsevier Inc. All rights reserved.)

An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method for screening of drugs in whole blood has been developed and validated. Samples were prepared by supported liquid-liquid extraction on ChemElute(®) columns with ethyl acetate/heptane (4:1). LC separation was achieved with an Acquity HSS T3-column (2.1 100 mm, 1.8-μm particle). Mass detection was performed by positive ion mode electrospray MS-MS and included the following drugs/metabolites: morphine, codeine, ethyl morphine, oxycodone, buprenorphine, methadone, cocaine, methylphenidate, amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), Δ(9)-tetrahydrocannabinol (THC), fentanyl, alprazolam, bromazepam, clonazepam, diazepam, nordiazepam, 3-OH-diazepam, fenazepam, flunitrazepam, lorazepam, nitrazepam, oxazepam, zopiclone, zolpidem, carisoprodol, and meprobamate. The cycle time was 9 min, and within- and between-day relative coefficients of variation varied from 1% to 33% and 2% to 58%, respectively. Extraction recoveries from whole blood were > 50% except for morphine and THC. The limit of quantitation was 0.1 to 521 ng/mL, depending on the drug.

Oral fluid is an alternative biological matrix that might have advantages over urine for drug analysis in treatment programs. A liquid chromatography-tandem mass spectrometry (LC-MS-MS) method has been used for screening 32 of the most commonly abused drugs and their metabolites in 0.5 mL preserved oral fluid, and the results were compared to results obtained from urine samples taken at the same time. In all, 164 pairs of oral fluid and urine were obtained from 45 patients stabilized on either methadone or buprenorphine. The total number of detections of drugs other than buprenorphine or methadone was 535 in oral fluid and 629 in urine. Morphine was found more often in urine (n = 66) than in oral fluid (n = 48), whereas the opposite was the case for 6-monoacetylmorphine (n = 20 in urine and n = 48 in oral fluid). Methadone showed the same detection frequency in urine and oral fluid (n = 75), whereas amphetamine (n = 45 in urine and n = 51 in oral fluid), methamphetamine (n = 39 in urine and n = 45 in oral fluid), and N-desmethyldiazepam (n = 37 in urine and n = 51 in oral fluid) were detected slightly more often in oral fluid. The other benzodiazepines, cannabis and cocaine were found more frequently in urine samples. If using a sensitive LC-MS-MS technique, oral fluid might be a good alternative to urine for detection of relatively recent use of drugs.

The aim of this study was to evaluate the stability of blood concentrations of a variety of illegal and medicinal drugs that are important for forensic analyses when spiked and stored in Vacutainer or Venosafe evacuated plastic collection tubes compared to Vacutainer evacuated glass tubes. Tubes were filled with spiked whole blood and analyzed after storage for one week at ambient temperature and at -20 degrees C, respectively. Freeze-and-thaw stability was included in the study. No significant difference between storage in glass or plastic tubes was noted for any compound investigated.

A liquid chromatography-tandem mass spectrometry (LC-MS-MS) method has been developed and validated for the determination of digoxin and digitoxin in whole blood samples in autopsy cases. Samples were prepared by liquid-liquid extraction (LLE) with ethyl acetate/heptane/dichloromethane (3:1:1). LC separation was achieved using an Atlantis dC(18)-column (2.1 x 50 mm, 3 microm). The time between injections was 11 min. Mass detection was performed by positive ion mode electrospray LC-MS-MS on the ammonium adducts with two transitions for each analyte and one for the internal standard (digoxin-d(3)). Within-day precision was between 8.3 and 10.8%, between-day precision was between 8.7 and 14.2% and accuracy (bias) was between -17.3 and 11.5%. LOQ was 0.1 nmol/L (0.08 ng/mL), with an accuracy and precision of 19% and -17% (digoxin) and 18% and 3% (digitoxin). Matrix effects ranged from 104 to 117%. Good qualitative correlation with previous findings was achieved for 38 autopsy cases. The median (range, number of cases) B-digitoxin and B-digoxin found with the LC-MS-MS method in this very limited material were 9.3 (3.4-23.8, n = 24) and 5.6 (3.4-26.5, n = 4) nmol/L, respectively.