Femoral blood concentrations of flualprazolam in 33 postmortem cases

Flualprazolam is a novel designer benzodiazepine, structurally related to alprazolam, flubromazolam and triazolam. In the last couple of years, it has been frequently detected in seizures and in forensic cases in Sweden and Finland. However, there is a lack of published blood concentrations for the drug, which presents difficulties when assessing its relevance for the cause of death. A quantitative method for the determination of flualprazolam in post-mortem blood was developed and validated, and subsequently used to analyse samples from 33 deaths previously screened as testing positive for flualprazolam in Sweden and Finland. Most of the cases in the study were accidental deaths (61 %) or suicides (18 %). The median (range) flualprazolam concentration was 18.0 (3.0-68) ng/g. The majority of the deceased were male (82 %) and the median age was 30 years. The median age in the Swedish cases was significantly higher (35 years) than in the Finnish cases (23 years) (p <0.05). Poly-drug use and particularly the concomitant use of flualprazolam and opioids were very common in the study population. Most of the cases that were positive for flualprazolam were fatal poisonings by a drug (N = 23), and in 13 cases, flualprazolam was implicated in the cause of death. Combining the resources of two countries in which all post-mortem toxicology is centralised provided a more comprehensive insight into the toxicology of flualprazolam. Research on novel psychoactive substances, such as flualprazolam, is required in order to be able to provide scientific evidence on the risks of these new substances for drug administration and potential users. (C) 2019 Elsevier B.V. All rights reserved.Peer reviewe

Enantioselective pharmacokinetics of tramadol and its three main metabolites; impact of CYP2D6, CYP2B6, and CYP3A4 genotype

Tramadol is a complex drug, being metabolized by polymorphic enzymes and administered as a racemate with the (+)- and (−)-enantiomers of the parent compound and metabolites showing different pharmacological effects. The study aimed to simultaneously determine the enantiomer concentrations of tramadol, O-desmethyltramadol, N-desmethyltramadol, and N,O-didesmethyltramadol following a single dose, and elucidate if enantioselective pharmacokinetics is associated with the time following drug intake and if interindividual differences may be genetically explained. Nineteen healthy volunteers were orally administered either 50 or 100 mg tramadol, whereupon blood samples were drawn at 17 occasions. Enantiomer concentrations in whole blood were measured by LC-MS/MS and the CYP2D6, CYP2B6 and CYP3A4 genotype were determined, using the xTAG CYP2D6 Kit, pyrosequencing and real-time PCR, respectively. A positive correlation between the (+)/(−)-enantiomer ratio and time following drug administration was shown for all four enantiomer pairs. The largest increase in enantiomer ratio was observed for N-desmethyltramadol in CYP2D6 extensive and intermediate metabolizers, rising from about two to almost seven during 24 hours following drug intake. CYP2D6 poor metabolizers showed metabolic profiles markedly different from the ones of intermediate and extensive metabolizers, with large area under the concentration curves (AUCs) of the N-desmethyltramadol enantiomers and low corresponding values of the O-desmethyltramadol and N,O-didesmethyltramadol enantiomers, especially of the (+)-enantiomers. Homozygosity of CYP2B6 *5 and *6 indicated a reduced enzyme function, although further studies are required to confirm it. In conclusion, the increase in enantiomer ratios over time might possibly be used to distinguish a recent tramadol intake from a past one. It also implies that, even though (+)-O-desmethyltramadol is regarded the enantiomer most potent in causing adverse effects, one should not investigate the (+)/(−)-enantiomer ratio of O-desmethyltramadol in relation to side effects without consideration for the time that has passed since drug intake

Smoking as a product of gene–environment interaction

A strong hereditary influence on smoking has been demonstrated. As one of the candidate genes in relation to smoking, the serotonin transporter gene (5-HTTLPR) has been suggested, however with conflicting results. In recent studies, it has been shown that genotypic and environmental (G*E) factors interact in the shaping of a variety of phenotypic expressions. The objective of the present study was to investigate the interaction between a variation in the 5-HTTLPR and family environment in relation to smoking habits, nicotine dependence, and nicotine and cotinine levels in hair samples

Studie av etanolens kinetik vid upprepade intag

Rapporten är framtagen med ekonomiskt bidrag från Trafikverket Skyltfonden. Ståndpunkter och slutsatser i rapporten reflekterar författaren och överensstämmer inte med nödvändighet med Trafikverkets ståndpunkter och slutsatser inom rapportens ämnesområde.Rattfylleri är ett stort trafiksäkerhetsproblem i Sverige. Årligen rapporteras dödsolyckor där den anhållne är misstänkt för grovt vållande till annans död, vårdslöshet i trafik, grovt rattfylleri samt avvikande från olycksplats. Särskilt problematiskt är om föraren efter olyckan lämnar platsen och vid påträffandet är alkoholpåverkad. Inte sällan uppger föraren då att hen druckit alkohol efter körningen, s.k eftersupning eller efterförtäring, vilket kan leda till bevissvårigheter för polis och åklagare. I de fall där den misstänkte inte anträffas i direkt anslutning till körningen är polisens rutiner att ta ett blodprov samt två urinprover med en timmas mellanrum. Genom detta förfarande kan man bedöma om det huvudsakliga alkoholintaget skett i nära anslutning till provtagningen eller längre tillbaka i tiden. Bedömningen utnyttjar skillnaden i blodalkoholhalt (BAC) och urinalkoholhalt (UAC) vid ett visst tillfälle samt skillnaden mellan två på varandra följande urinprovers UAC. En UAC/BAC kvot högre än 1,25 indikerar att all alkohol har fördelats i kroppen och att man är i eliminationsfasen. En kvot mindre än 1,0 talar starkt för att man nyligen intagit alkohol och fortfarande är i absorptionsfasen. Den andra parametern är UAC2-UAC1 som visar sjunkande värde om man är i eliminationsfasen. Det vetenskapliga underlaget för dessa parametrar är hämtat från experimentella studier med engångsintag av alkohol, vanligtvis starksprit under en kort period. Förhållandet mellan alkoholhalten i blod och urin kompliceras emellertid när alkoholförtäringen utsträckts över en längre tidsrymd eller om man även druckit alkohol vid ett tidigare tillfälle. Man kan anta att merparten av de personer som observerats dricka alkohol efter körningen också har gjort det före färden. På vilket sätt detta scenario skulle kunna inverka på alkoholkurvorna i blod och urin är inte utrett. I den aktuella studien gavs därför alkohol vid två olika tillfällen och i tre doser i form av antingen öl, vin eller starksprit. I Norge används en annorlunda modell för att utreda efterförtäring, nämligen analys av etanolens metaboliter etylglukuronid (EtG) och etylsulfat (EtS). Vi avsåg därför även att jämföra den svenska och norska modellen i vår studie. Våra hypoteser att halter och förhållandet mellan etanol i blod och urin liksom av EtG och EtS i blod kan användas för att bedöma när en person senast intagit etanol visade sig sanna under de nya förhållandena och bekräftar att båda modellerna är användbara även när den misstänkte har alkohol i blodet sedan tidigare. Kvoten mellan halten etanol och halten EtG i ett blodprov är den parameter som bäst kan upptäcka om ett intag skett i nära anslutning till provtagningarna särskilt om efterintaget är en liten mängd alkohol. Vi bedömer det som mycket osannolikt att efterförtäring av stora mängder alkohol skulle kunna förbises även om den misstänkte haft alkohol i kroppen vid tillfället för eftersupningen oavsett vilken modell som används

Förslag till förbättringar av rattfylleristatistiken i Sverige : underlag till uppdrag att förbättra beskrivningen av vägtrafikolyckornas hälsopåverkan

In June 2006, the Swedish Road Administration was mandated by the Government to improve the description of the health effects of road traffic accidents. Under the terms of this mandate, a proposal was to be made as to how the recording of alcohol and drug related accidents can be improved, and how sample surveys of the prevalence of drivers under the influence of alcohol and/or drugs should be performed. This note describes the data used for these parts of the mandate. One common condition for both these parts is that possible screening methods for drivers under the influence of narcotics should be studied. In order to measure the proportion of traffic mileage travelled by drink/drug drivers, the investigation should be organised as a statistical sample survey where the police check drivers at sites selected at random. As regards selection in time, it may be reasonable to use quota sampling. In the present situation, we do not consider that it is possible to carry out regular surveys of drug driving. If the regulations are amended so that routine controls can be performed, and if the police begin to use screening instruments for drug tests, regular surveys can be performed. These surveys can then be integrated with the investigations regarding alcohol, by requiring the stopped drivers to give samples for both alcohol and for drugs

First evaluation of the possibility of testing for drugged driving using exhaled breath sampling

Objective: Driving under the influence of psychoactive drugs causes an increased risk for accidents. In combating this, sobriety tests at the roadside are common practice in most countries. Sampling of blood and urine for forensic investigation cannot be done at the roadside and poses practical problems associated with costs and time. An alternative specimen for roadside testing is therefore warranted and the aerosol particles in exhaled breath are one such alternative.Methods: The present study investigated how the exhaled breath sample compared with the routine legal investigations of blood and urine collected from suspects of drugged driving at 2 locations in Sweden. Exhaled breath was collected using a simple filter collection device and analyzed with state-of-the-art mass spectrometry technique.Results: The total number of cases used for this investigation was 67. In 54 of these cases (81%) the results regarding a positive or negative drug test result agreed and in 13 they disagreed. Out of these, the report from the forensic investigation of blood/urine was negative in 21 cases. In 6 of these, analytical findings were made in exhaled breath and these cases were dominated by the detection of amphetamine. In 7 cases a positive drug test from the forensic investigation was not observed in the breath sample and these cases were dominated by detection of tetrahydrocannabinol in blood. In total, 45 samples were positive with breath testing and the number of positives with established forensic methods was 46.Conclusion: The promising results from this study provide support to exhaled breath as a viable specimen for testing of drugged driving. The rapid, easy, and convenient sampling procedure offers the possibility to collect a drug test specimen at the roadside. The analytical investigation must be done in a laboratory at present because of the need for a highly sensitive instrument, which is already in use in forensic laboratories. The analytical work is not more challenging than for blood or oral fluid and should not cause an increase in cost. However, more studies need to be done before exhaled breath drug testing can be applied routinely for drugged driving investigation

Characterization of neurotransmitter inhibition for seven cathinones by a proprietary fluorescent dye method

Many new psychoactive substances (NPS) are stimulants, and information about their potency and abuse potential is often lacking. To start addressing this need, a method measuring the inhibition of the dopamine, serotonin, and norepinephrine transporters (DAT, SERT, and NET) by stimulant drugs was developed. The use of a proprietary fluorescent dye mixture and three cell lines (CHO-K1, HEK 293, and MDCK), each expressing a single transporter, allowed for a semiautomated, one-pot determination of inhibition in a 384-well format. The method was validated using well characterized stimulants, including cocaine, amphetamine, 3,4-methylenedioxymethamphetamine (MDMA), &amp; alpha;-PVP, and fluoxetine and performed similarly to other methods. Seven synthetic cathinones all showed highest potency for DAT inhibition, followed by NET and SERT. The rank potency for DAT inhibition IC50 (nM) was MPHP (4.53) &amp;gt; 4Cl-&amp; alpha;-PVP (8.05) &amp;gt; 3F-&amp; alpha;-PVP (12.7) &amp;gt; &amp; alpha;-PiHP (13.4) &amp;gt; N-ethylpentylone (16.9) &amp;gt; N-ethylhexedrone (44.5) &amp;gt; 4-methylpentedrone (261). All but 4-methylpentedrone were more potent than amphetamine (257) and cocaine (111). The DAT/SERT inhibition ratio for the cathinones was in the range from 5.02 for 4-methylpentedrone to &amp;gt;3730 for &amp; alpha;-PiHP, compared to 1.64 for cocaine and &amp;gt;4030 for &amp; alpha;-PVP. All seven substances had inhibition profiles similar to those of potent stimulants with high abuse potential.Funding Agencies|Public Health Agency of Sweden; National Board of Forensic Medicine</p

In vitro characterization of new psychoactive substances at the mu-opioid, CB1, 5HT(1A), and 5-HT2A receptors-On-target receptor potency and efficacy, and off-target effects

New psychoactive substances (NPS) appear on the recreational market on a monthly basis, with unclear toxicology, resulting in an increasing number of fatalities. Identification of drug targets and potencies is crucial for understanding and treating intoxications and for scheduling processes. In this study 60 NPS and metabolites belonging to opioids, cannabinoids and serotonergic hallucinogens classes were screened for in vitro activation of the mu-opioid, CB1, 5-HT1A and 5-HT2A receptors using the AequoZen cell system. Fentanyl and NBOMe analogues were chosen for full dose-response characterization of the mu-opioid and 5-HT2A receptors, respectively. Most substances activated their corresponding target receptor. The most potent mu-opioid receptor agonists were 2-fluorofentanyl (EC50 = 1.0 nM), carfentanil (EC50 = 2.7 nM) and acrylfentanyl (EC50 = 2.8 nM) and in total a &amp;gt;1500-fold difference was seen among the tested compounds. Moreover, furanylfentanyl, 4-methoxybutyrylfentanyl and valerylfentanyl acted as partial agonists of the mu-receptor. On the 5-HT2A receptor, bromo-dragonfly showed the highest potency (EC50 = 0.05 nM, 400 times more potent than LSD), followed by most NBOMe compounds with EC50 values ranging from 0.11 nM (for 25N-NBOMe) to 1.3 nM (for 25T4-NBOMe)). Off-target activation of the mu-opioid receptor was identified for piperazines, phenethylamines (in particular NBOMe and 2C compounds) and tryptamines. Moreover, the synthetic cannabinoid metabolite 3-carboxy indole PB-22 activated the 5-HT2A receptor. Bromo-dragonfly was the only compound that activated all four receptors. These results highlight the possible interplay of known and unknown NPS targets and unveil its complexity. Moreover, the detailed, quantitative information presented facilitates our further understanding of NPS toxicology. (c) 2020 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).Funding Agencies|Strategic Research Area in Forensic Sciences at Linkoping University [2016:6]</p

A Rapid Method for Postmortem Vitreous Chemistry - Deadside Analysis

Vitreous fluid is commonly collected for toxicological analysis during forensic postmortem investigations. Vitreous fluid is also often analyzed for potassium, sodium, chloride and glucose for estimation of time since death, and for the evaluation of electrolyte imbalances and hyperglycemia, respectively. Obtaining such results in the early phase of a death investigation is desirable both in regard to assisting the police and in the decision-making prior to the autopsy. We analyzed vitreous fluid with blood gas instruments to evaluate/examine the possible impact of different sampling and pre-analytical treatment. We found that samples from the right and left eye, the center of the eye as well as whole vitreous samples gave similar results. We also found imprecision to be very low and that centrifugation and dilution were not necessary when analyzing vitreous samples with blood gas instruments. Similar results were obtained when analyzing the same samples with a regular multi-analysis instrument, but we found that such instruments could require dilution of samples with high viscosity, and that such dilution might impact measurement accuracy. In conclusion, using a blood gas instrument, the analysis of postmortem vitreous fluid for electrolytes and glucose without sample pretreatment produces rapid and reliable results

Drug and medicine prevalence in traffic in Sweden : the results within the EU project DRUID

The prevalence of illicit drugs and medicines in the driving population, i.e. among drivers on the road, was 2.5 per cent between the years 2006 and 2011 in Sweden. The prevalence of alcohol, illicit drugs and medicines among drivers killed in traffic the same years was 31.2 per cent. In comparison with other countries in the EU project DRUID, Sweden has the highest proportion of negative results, i.e. drivers without the presence of alcohol, drugs or medicine. This applies to both drivers on the road and drivers killed in traffic. The studies that form the basis of this report have been performed within the project DRUID - Driving under the influence of drugs, alcohol and medicine, which is a project within the EU's Sixth Framework Programme, which lasted five years (2006-2011). This report presents two sub-studies from the DRUID project. The aims of these studies were: - to study the prevalence of illicit drugs and medicines in the driving population (study among drivers on the road). - to study the prevalence of alcohol, illicit drugs and medicines among killed drivers (study of killed drivers). In the study among drivers on the road, saliva samples were collected. Toxicological results were analysed from 6,199 drivers in Södermanland, Örebro and Östergötland out of 10,223 drivers of passenger cars and vans that were stopped by the police in March 2008- February 2009. In the study of killed drivers we have toxicological results from 157 fatalities out of 178 drivers of passenger cars and vans that were killed in traffic accidents in 2008