J.-M. Gaulier

Detection of the designer benzodiazepine metizolam, in urine and preliminary data on its metabolism

Designer benzodiazepines provide an attractive alternative to prescribed benzodiazepines for abuse purposes as they are readily available via the Internet without control. Metizolam was ordered via the Internet and a 2u2009mg blue tablet was orally administered to a 54-year-old man. Urine samples were collected over 6u2009days in polypropylene tubes. After liquid/liquid extraction at pHu20099.5, metizolam was analyzed by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) using a standard method devoted to benzodiazepines, and ions transitions, at m/z 328.9u2009>u2009275.0 and 328.9u2009>u2009300.0. Metizolam was detectable in hydrolyzed urine during the 46-h period, with concentrations always lower than 11u2009ng/mL. About 0.3% of the initial dose was excreted in urines as total unchanged metizolam during the first 24u2009h. The most relevant potential CYP- and UGT-dependent metabolites of metizolam were investigated in vitro using human liver microsome incubation and, subsequently, liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF-MS) analysis. Three mono-hydroxylated metabolites were produced including a hydroxylation compound at the 2-ethyl moiety of metizolam (M1) as quantitatively main metabolite, and a N-hydroxymetiazolam (M2). The structure of the third metabolite (M3) could not be elucidated because of a too low experimental production rate. Two authentic urine samples were analyzed using the same analytical method to search for metabolites of metizolam. M1, together with its glucuronide (M1-Glu), and M2 were observed in urine at the 8u2009h mark, whereas only M1 and M1-Glu were still detected in urine at 30u2009h post administration. Copyright

Prevalence of New Psychoactive Substances and Prescription Drugs in the Belgian Driving under the Influence of Drugs Population

Driving under the influence of drugs (DUID) is a worldwide problem. Several countries have adopted DUID legislations which prove their deterrent effect and impact on road safety. However, the use of new psychoactive substances (NPS) and prescription drugs is not known, as the applied roadside screening tests have not yet been adapted for these compounds. In this study, 558 blood samples obtained during roadside controls in Belgium (January to August 2015) after a positive Drugwipe 5S® test and 199 oral fluid (OF) samples obtained from negatively screened test pads were analyzed. The NPS positivity rate was 7% in blood, while it reached 11% in OF. NPS detected were: diphenidine, ketamine, 4-fluoroamphetamine, 2-amino-indane, methoxetamine, α-PVP, methiopropamine, a mix of 5-MAPB/5-EAPB, TH-PVP, mephedrone, methedrone, 4-methylethylcathinone, 5-MeO-DALT, 4-Acetoxy-DiPT, AB Fubinaca, FUB-JWH018, JWH020, trifluoromethylphenylpiperazine, and ethylphenidate. Moreover, 17% of blood samples (and 5% of OF) contained an analgesic drug, 10% (0.5%) a benzodiazepine/hypnotic, 5% (2%) an antidepressant, 2% (3%) an antipsychotic, 2% an antiepileptic drug, and 1% methylphenidate. The presence of NPS in the young (and predominately male) DUID population is proven. Furthermore, a high level of poly-drug use including combinations of NPS, licit, and drugs of abuse was observed. Further research concerning the development of on-site NPS detection techniques should be established. Meanwhile, the effects of combined drug use on driving ability and the physical/psychological signs after NPS use should be performed to improve the on-site DUID detection of NPS by police officers, so they can engage in blood sampling for a general unknown screening.

Fatality involving ocfentanil documented by identification of metabolites

The use of new psychoactive substances (NPS) has rapidly increased over the last decade. In the last 4xa0years, producers increasingly appear to be targeting non-controlled synthetic opioids, involving fentanyl derivatives such as ocfentanil (OcF). Identification of metabolites is of major importance in the context of NPS use, as it could improve the detection window in biological matrices in clinical and forensic intoxication cases. Hence, this work aims to report a fatality involving OcF documented by the identification of metabolites. A 30-year-old woman was found dead at home: an unidentified powder was found near her body and some injection sites were found at the autopsy. Toxicological analyses allowed to determine the presence of OcF in the powder, blood (3.7/3.9xa0μg/L, peripheral/cardiac) and in other post-mortem samples. The most relevant potential CYP- and UGT-dependent metabolites of OcF were investigated in vitro using human liver microsome incubation and liquid chromatography coupled with high resolution mass spectrometry, and subsequently confirmed in post-mortem samples. Four OcF metabolites were produced in vitro (a mono-hydroxylated OcF, O-desmethylOcF, a hydroxylated desmethylOcF and a glucuronidated form of the O-desmethylOcF), and all except the glucuronide were observed in blood and bile post-mortem samples. Considering the relative intensity of the chromatographic peak areas, O-desmethylOcF can be suggested to be an abundant metabolite of OcF. Nevertheless, the relevance of O-desmethylOcF as being a complementary analytical target of OcF for OcF use detection needs further in vivo confirmation, especially through analysis of urines from users.

Unusual Case of Drug-Facilitated Sexual Assault using Chloroform

Despite its well-known sedative properties, chloroform is rarely used for drug-facilitated sexual assault (DFSA) as its administration cannot be achieved without the victims knowledge: we report an unusual case of DFSA using this solvent. A 26-year-old woman declared that her partner get her to sleep using chloroform the previous night. When she waked up at 3 am, her hands were tied. She immediately suspected violence and sexual penetration. Toxicological blood screening using a liquid chromatography-electrospray coupled tandem mass spectrometry method highlighted the presence of bamifylline and theophylline, two therapeutics of asthma. A screening method for volatile substances using a headspace-gas chromatography coupled with mass spectrometry method showed (i) the presence of chloroform in blood at a concentration subsequently estimated at 580 µg/L using an external calibration, and (ii) chloroform traces on a piece of a scarf brought by the patient and suspected to have been used to put her to sleep. These results were consistent with an exposure to chloroform by inhalation and demonstrate that there is no limit in the use of chemical weapons in DFSA.

Screening en 24/24 en toxicologie hospitalière : comparaison des résultats entre chromatographie liquide — spectrométrie UV à barrette de diodes (LC-DAD) associée à l’immunoanalyse vs chromatographie liquide — trappe ionique (LC–SM n )

Objectif Appliquer une methode separative associant la chromatographie liquide a une detection par spectrometrie de masse de type trappe ionique dans le cadre du screening toxicologique pour les urgences hospitalieres et la comparer a la strategie «xa0de routinexa0» en 24/24xa0du laboratoire (chromatographie liquide couplee a une barrette de diodes (LC-DAD)xa0±xa0immunoanalyses (IA) afin d’evaluer les avantages et inconvenients de ce nouveau systeme de criblage. Methodes Cent quarante-sept echantillons urinaires de patients admis aux urgences ou en reanimation ont ete analyses a l’aide du Toxtyper™ (LC-SM n , Bruker) et de notre strategie de screening urinaire combinant LC-DADxa0±xa0IA (stupefiantsxa0±xa0traitement de substitution aux opiaces [TSO]xa0±xa0benzodiazepines [BZD]). Les methodes de preparation des echantillons et les librairies (temps de retention et spectres UV ou de masse) sont celles fournies par les constructeurs. Resultats Aucun xenobiotique n’a ete detecte dans deux urines quelle que soit la methode utilisee. Au moins un xenobiotique (medicament ou stupefiant) a ete detecte dans 143/145xa0echantillons par le Toxtyper™ et dans 97/145xa0echantillons par la LC-DAD. 129/145xa0echantillons ont ete detectes positifs pour au moins un stupefiant, une BZD ou un TSO par IA. La comparaison des resultats obtenus ( n xa0=xa0nombre d’urines detectees) via le Toxtyper™ et notre strategie de screening urinaire est presentee par classe de molecules dans le Tableau 1 . Discussion Une ou plusieurs molecules d’interet toxicologique ont ete detectees dans 98,6xa0% des 145xa0echantillons urinaires par le Toxtyper™ et dans 67xa0% des echantillons par la LC-DAD avec ici un reel interet de la combinaison LC-DADxa0+xa0IA, notamment pour la detection des stupefiants et des BZD. Des essais d’optimisation de la preparation d’echantillon montrent une amelioration de l’efficacite du Toxtyper™. Par exemple, pour les 10xa0echantillons initialement discordants pour les BZD ( * ), l’utilisation de notre preparation d’echantillon permet au Toxtyper™ d’en detecter la presence. Conclusion Le Toxtyper™ permet l’identification de nombreuses molecules d’interet toxicologique et/ou de leurs metabolites dans un delai compatible avec l’urgence. A ce titre, le Toxtyper™ trouve toute sa place dans une strategie de screening en toxicologie biologique (toxicologie hospitaliere) en 24/24.

New psychoactive substances in oral fluid of French and Belgian drivers in 2016

BACKGROUNDnDriving under the influence of drugs (DUID) is a worldwide problem with potentially major judiciary and life-threatening consequences. Up to now, only classical drugs of abuse (DOA) are tested for DUID detection. A challenging issue for drafting up-dated international drug policies is to take into account the recent and expanding new psychoactive substances (NPS) market. NPS consist in various narcotic or psychotropic drugs, most of them having a legal status, that replicate chemical structures and/or pharmacological effects of classical DOA. Although it is obvious that NPS can lead to impaired driving, the prevalence of NPS use in a DUID context is unknown since the applied roadside screening tests are not yet adapted for these compounds.nnnMETHODSnBetween January and December 2016, a total of 391 oral fluid specimens were obtained from used roadside immunochemical test devices for DOA (Drugwipe-5S® device). These specimens were analyzed using liquid chromatography coupled with tandem mass spectrometry and high resolution mass spectrometry.nnnRESULTSnNPS (mainly cathinone derivatives) were detected in 33 out of the 391 oral fluid samples. This NPS positivity rate of 8.4% in oral fluid of drivers who were submitted to a roadside drug testing in 2016 in France and in Belgium is comparable to the available blood data (NPS positivity rate of 7%) observed in 2015 in similar populations.nnnCONCLUSIONnOur results demonstrate the reality of driving after NPS use in French and Belgian drivers who were submitted to a roadside DOA test. As there is a lack of on-site detection methods to screen for NPS, the detection of NPS in a rapid and cost-effective DUID detection strategy is currently impossible. The expanding use of NPS, notably by drivers as reported here, and the inability of currently used drug detection tests, should be urgently addressed by road safety and law enforcement authorities.