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Objectives A method based on liquid chromatography coupled to triple quadrupole mass spectrometry detection using 50 µL of plasma was developed and fully validated for quantification of remdesivir and its active metabolites GS-441524. Methods A simple protein precipitation was carried out using 75 µL of methanol containing the internal standard (IS) remdesivir-13C6 and 5 µL ZnSO4 1 M. After separation on Kinetex® 2.6 µm Polar C18 100A LC column (100 × 2.1 mm i.d.), both compounds were detected by a mass spectrometer with electrospray ionization in positive mode. The ion transitions used were m/z 603.3 → m/z 200.0 and m/z 229.0 for remdesivir, m/z 292.2 → m/z 173.1 and m/z 147.1 for GS-441524 and m/z 609.3 → m/z 206.0 for remdesivir-13C6. Results Calibration curves were linear in the 1–5000 μg/L range for remdesivir and 5–2500 for GS-441524, with limit of detection set at 0.5 and 2 μg/L and limit of quantification at 1 and 5 μg/L, respectively. Precisions evaluated at 2.5, 400 and 4000 μg/L for remdesivir and 12.5, 125, 2000 μg/L for GS-441524 were lower than 14.7% and accuracy was in the [89.6–110.2%] range. A slight matrix effect was observed, compensated by IS. Higher stability of remdesivir and metabolite was observed on NaF-plasma. After 200 mg IV single administration, remdesivir concentration decrease rapidly with a half-life less than 1 h while GS-441524 appeared rapidly and decreased slowly until H24 with a half-life around 12 h. Conclusions This method would be useful for therapeutic drug monitoring of these compounds in Covid-19 pandemic.

International audience; The aim of this study was to identify new markers of deschloro-N-ethyl-ketamine (O-PCE), a ketamine analogue that has been involved in acute intoxications with severe outcomes including death and whose metabolism has never been studied before. In vitro study after 2-h incubation with pooled human liver microsomes (HLMs) cross-checked by the analysis of urine and hair from a 43-year-old O-PCE user (male) were performed by liquid chromatography–high resolution mass spectrometry (LC-HRMS). Acquired data were processed by the Compound Discoverer® software, and a full metabolic profile of O-PCE was proposed. In total, 15 metabolites were identified, 10 were detected in vitro (HLMs) and confirmed in vivo (urine and/or hair), two were present only in HLMs, and the remaining three metabolites were identified only in biological specimens. While O-PCE was no longer detected in urine, nine metabolites were identified allowing to increase its detection window. In descending order of metabolites abundance, we suggest using 2-en-PCA-N-Glu (34%, first), M3 (16%, second), O-PCA-N-Glu (15.4%, third), OH-O-PCE (15%, fourth) and OH-PCE (11.9%, fifth) as target metabolites to increase the detection window of O-PCE in urine. In hair, nine metabolites were identified. OH-PCA was the major compound (78%) with a relevant metabolite to parent drug ratio (=6) showing its good integration into hair and making it the best marker for long-term monitoring of O-PCE exposure.

Objective To develop a population pharmacokinetic model for lopinavir boosted by ritonavir in coronavirus disease 2019 (Covid-19) patients. Methods Concentrations of lopinavir/ritonavir were assayed by an accredited LC-MS/MS method. The population pharmacokinetics of lopinavir was described using non-linear mixed-effects modeling (NONMEM version 7.4). After determination of the base model that better described the data set, the influence of covariates (age, body weight, height, body mass index (BMI), gender, creatinine, aspartate aminotransferase (AST), alanine aminotransferase (ALT), C reactive protein (CRP), and trough ritonavir concentrations) was tested on the model. Results From 13 hospitalized patients (4 females, 9 males, age = 64 ± 16 years), 70 lopinavir/ritonavir plasma concentrations were available for analysis. The data were best described by a one-compartment model with a first-order input (KA). Among the covariates tested on the PK parameters, only the ritonavir trough concentrations had a significant effect on CL/F and improved the fit. Model-based simulations with the final parameter estimates under a regimen lopinavir/ritonavir 400/100 mg b.i.d. showed a high variability with median concentration between 20 and 30 mg/L (Cmin/Cmax) and the 90% prediction intervals within the range 1–100 mg/L. Conclusion According to the estimated 50% effective concentration of lopinavir against SARS-CoV-2 virus in Vero E6 cells (16.7 mg/L), our model showed that at steady state, a dose of 400 mg b.i.d. led to 40% of patients below the minimum effective concentration while a dose of 1200 mg b.i.d. will reduce this proportion to 22%. Electronic supplementary material The online version of this article (10.1007/s00228-020-03020-w) contains supplementary material, which is available to authorized users.

International audience; This study was to validate a LC–MS/MS method for the determination of 17 new synthetic opioids (NSOs) in hair including 3-fluorofentanyl, 3-methylfentanyl, acetylfentanyl, acetylnorfentanyl, alfentanyl, butyrylfentanyl, butyrylnorfentanyl, carfentanil, fentanyl, furanylfentanyl, furanylnorfentanyl, methoxyacetylfentanyl, norcarfentanil, norfentanyl, ocfentanil, sufentanil, and U-47700, and to apply it to 137 authentic samples. Twenty milligrams of hair was decontaminated in dichloromethane and underwent liquid extraction. 10 μL of the reconstituted residue were injected onto the system. The separation was performed in 12 minutes in a gradient mode at a flow rate of 300 μL/min using a Hypersyl Gold PFP column (100 × 2.1 mm i.d., 1.9 μm) maintained at 30°C. Compounds were detected in positive ionization and MRM modes using a TSQ Endura mass spectrometer (ThermoFisher). The method was validated according to EMA guidelines. The LLOQ was in the range 1–50 pg/mg, and the calibration ranged from the LLOQ-1000 pg/mg. Intra- and inter-day accuracy (bias) and precision were < 15%. Extraction recoveries of parent drugs and metabolites were 74–120% and 7–62%, respectively. The matrix effect was in the range 59–126% (CVs ≤ 12.9%). Fentanyl was found in six cases at concentrations of < 1–1650 pg/mg (n = 14 segments). Five fentanyl analogs were quantified in two cases: 3-fluorofentanyl (25–150 pg/mg, n = 5), furanylfentanyl (15–500 pg/mg, n = 5), methoxyacetylfentanyl (500–600 pg/mg, n = 2), acetylfentanyl (1 pg/mg, n = 2), carfentanyl (2.5–3 pg/mg, n = 2). This fully validated method allowed us to test for the first time 3-fluorofentanyl and norcarfentanil in hair among 15 other NSOs, and brings new data regarding 3-fluorofentanyl and methoxyacetylfentanyl hair concentrations.

Untargeted liquid chromatography-high resolution mass spectrometry (LC-HRMS) techniques have become indispensable tools for systematic toxicological analysis. They allow the research of an almost unlimited number of drugs within a single analytical cycle, but shared mass spectra libraries are still missing to identify newly marketed compounds, along with defined analytical procedures. This article describes the optimization, validation, and application of an untargeted screening method devoted to hair analysis using data-dependent analysis (DDA) and a shared HRMS database. This method used an ultra-high performance liquid chromatography coupled to a benchtop Orbitrap. Raw MS data were processed with Compound Discoverer software coupled to the mzCloud™ library. Optimizations were performed on blank hair spiked with 19 analytes having different physical and chemical properties. To validate the effectiveness of a shared spectra database, 20 compounds spectra were added and then retrospectively screened. Sensitivity and reliability were evaluated on 317 compounds of interest in toxicology. The method was then applied to 11 hair samples. The matrix effect range by ion suppression/enhancement was 40%-110%. The method allows the detection of 284 among the 317 screened compounds, including 72 new psychoactive substances (NPS). Lower limit of identification (LLOI) and lower limit of detection (LLOD) were 1 to 1000 pg/mg and 1 to 500 pg/mg, respectively. The method was successfully applied to 11 clinical cases and 144 compounds were identified including 24 NPS including AKB48-5F for the first time in hair. We developed and validated an LC-HRMS untargeted screening of 284 compounds and successfully applied it to 11 real hair samples.

Recently, the emergence of new psychoactive substances (NPS) has led to their wide use among clubbers and men who have sex with men (MSM) for their stimulant effects. However, their use in drug-facilitated sexual assault (DFSA) has rarely been described. Herein we report a case of a 44-year-old man who was assaulted after a party. Due to late reporting of the offense, only hair (black) was sampled 15 days later and a segmental analysis was achieved to look for most DFSA agents and NPS. Twenty mg of each segment (A: 0-1 cm, B: 1-3 cm, and C: 3-5 cm) were incubated in phosphate buffer pH 5.0. After alkaline liquid extraction and chromatographic separation on 1.9 μm Hypersil GOLD PFP column, compounds were detected by a TSQ Vantage mass spectrometer with electrospray ionization in positive mode with multiple reaction monitoring (MRM) acquisition. 4-methylethcathinone (4-MEC), methylenedioxypyrovalerone (MDPV) and doxylamine were found in proximal segment at very low concentrations (3, 5, and 9 pg/mg, respectively) which is in agreement with a single exposure in the previous month corresponding to the alleged facts. These substances were not detected in segments B and C showing a lack of repetitive exposure before the alleged event. Thus, the results do not contradict the patient's claim of being assaulted. Doxylamine has already been encountered in such cases but no publications referring to 4-MEC or MDPV use have ever been documented. Our case reports the unusual administration of cathinones to achieve a sexual assault and stresses the interest of looking for designer drugs when dealing with DFSA cases.

Tranexamic acid is a widely used antifibrinolytic drug but its pharmacology and pharmacokinetics remains poorly understood. Owing to the recent knowledge on phospholipid-induced matrix effects during human plasma analysis, our aim was to develop a liquid chromatography-mass spectrometry method for the quantitation of tranexamic acid after efficient sample clean-up. Sample preparation consisted in phospholipid removal and protein precipitation. Hydrophilic interaction liquid chromatography was used and the detection was achieved with multiple reaction monitoring. The method was validated according to the European Medicine Agency guideline in the range 1.0–1000.0 μg/mL. The performance of the method was excellent with a precision in the range 1.2-3.0%, an accuracy between 88.4 and 96.6% and a coefficient of variation of the internal standard-normalized matrix factor below 6.7%. This method is suitable for the quantification of tranexamic acid in the wide range of concentrations observed during clinical studies, with all the advantages related to phospholipid removal.

Resume Objectif La fabrication et la vente illegale de peptides recombinants deviennent de plus en plus facile et frequente. Nous rapportons ici le cas d’une poudre blanche etiquetee comme etant du CJC-1295, un derive d’un analogue peptidique synthetique de la GHRH, analysee afin d’en determiner la composition reelle. Methode Un echantillon de poudre a ete analyse sur un systeme CL-SM/SMHR de type Q-Exactive (Thermo, Etats-Unis). L’instrument fonctionnait en mode data-dependent ( full scan suivi d’une fragmentation des ions d’abondance significative) a une resolution de 70 000 avec une ionisation de type electrospray fonctionnant en mode positif. Resultats Le chromatogramme a montre que la poudre etait pure. L’analyse spectrale a mis en evidence 5 ions multicharges. L’ion monoisotopique le plus abondant etait le m/z 674,1855 ( z = 5) correspondant a une masse de 3365,8885 Da. La masse observee correspondait a l’analogue peptidique de la GHRH de 29 acides amines contenu dans le CJC-1295 (3365,8930 Da) avec une erreur de masse de 1,3 ppm. Cet analogue peptidique n’etait pas conjugue contrairement a celui present dans le CJC-1295. Conclusion L’interpretation du spectre SMHR associe a la connaissance de la structure de la molecule nous a permis d’identifier l’analogue peptidique contenu normalement dans le CJC-1295, sans pour autant etre du CJC-1295 dans lequel cet analogue est couple a un complexe qui en prolonge la demi-vie. Il s’agit a notre connaissance de la premiere saisie de ce produit en France.

Dear Editor,Hyperemesis has been described in tetrahydrocannabinol (THC) and/or synthetic cannabinoid users, but not with cannabidiol (CBD) [1]. We report a case of hyperemesis associated with CBD ...

Purpose To demonstrate the usefulness of hair testing to determine exposure pattern to fentanyls. Methods A 43-year-old male was found unconscious with respiratory depression 15 min after snorting 3 mg of a powder labeled as butyrylfentanyl. He was discharged from hospital within 2 days without blood or urine testing. Two locks of hair were sampled 1 month (M1 A: 0–2 cm (overdose time frame); B: 2–4 cm; C: 4–6 cm) and 1 year (Y1: A: 0–2 cm; B: 2–4 cm) later to monitor his exposure to drugs of abuse by liquid chromatography–tandem mass spectrometry after liquid-liquid extraction. Results Hair analysis at M1 showed a repetitive exposure to 3-fluorofentanyl (A/B/C: 150/80/60 pg/mg) with higher concentration in segment A reflecting the overdose period. The non-detection of butyrylfentanyl was consistent with the analysis of the recovered powder identified as 3-fluorofentanyl. Furanylfentanyl (40/20/15 pg/mg) and fentanyl (37/25/3 pg/mg) were also detected in hair. The second hair analysis at Y1 showed the use of three new fentanyls, with probably repetitive exposures to methoxyacetylfentanyl (A/B: 500/600 pg/mg), and single or few exposures to carfentanil (2.5/3 pg/mg) and acetyl fentanyl (1/1 pg/mg). A decreasing exposure to 3-fluorofentanyl (25/80 pg/mg), and increasing consumption of furanylfentanyl (310/500 pg/mg) and fentanyl (620/760 pg/mg) were also observed despite methadone treatment initiation. The patient claimed not consuming three out of the six detected fentanyls. Conclusions We report single or repetitive exposure to several fentanyls using hair testing. To our knowledge, this is the first demonstration of 3-fluorofentanyl and methoxyacetylfentanyl in hair samples collected from an authentic abuser.

Summary Purpose Synthetic cannabinoids (SCs) are the largest group of new psychoactive substances (NPS) monitored worldwide. We validate a rapid and sensitive liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) assay for the determination of 16 SCs in human hair samples. This method was successfully applied to document chronic use of JWH-122 in a 42 years-old male included in large-scale cross-sectional study in Paris. Method The SCs JWH-018, JWH-018-5-N(OH-pentyl), JWH-073, JWH-073-4-N(OH-butyl), JWH-122, JWH-200, JWH-019, JWH-250, AM-2201, EAM-2201, BB-22, ADB-PINACA, AKB48-5F, AB-FUBINACA, 5F-AB-PINACA, 5F-PB-22 were extracted from 20 mg of decontaminated, washed and grinded hair by a mixture of hexane/ethyl acetate (1/1, v/v) in alkaline condition in the presence of JWH-018-D11 internal standard. SCs were then separated by a gradient of formate buffer 2 mM in 0.1% formic acid (A) and acetonitrile (B) on Hypersyl Gold PFP column (100 × 2.1 mm i.d., particle size 1.9 µm, ThermoFisher) at 30 °C. The detection was performed on TSQ Vantage triple-quadrupole mass spectrometer (Thermofisher, Les Ulis, France) in positive ionization and multi-reaction monitoring (MRM) modes. A lock of hair (6 cm) was sampled from a patient hospitalized for JWH 122 overdose and segmented into 3 segments: A: 0-2 cm (proximal), B: 2-4 cm and C: 4-6 cm. It was then analyzed with the present method in order to determine past use of synthetic cannabinoids. Results The method exhibits a good linearity (r2 > 0.996) over the range 1-25 to 1000 pg/mg, Limits of detection and quantification ranged from 0.3 to 8 pg/mg and from 1 to 25 pg/mg, respectively (CVs ≤ 15%). No interference from endogenous substances with the detection of SCs and their IS was observed and the carryover was insignificant. Intraday and inter-day trueness (bias) and precision (CVs) were ≤ 8.2%, ≤ 14.3%, ≤ 8.3% and ≤ 14.9%, respectively. Overall recoveries ranged between 61 and 94% and corrected matrix effect from 31 to 110%. JWH-122 hair concentrations found in patient's hair were: (A/B/C: 200/450/430 pg/mg) in agreement with the concentrations found in the literature suggesting chronic use of JWH-122 over the last six months. Conclusion The present LC-MS/MS method proved to be suitable for the detection and quantification of 16 synthetic cannabinoids in human hair.

Summary Adipocere formation requires very particular conditions of humidity and pH, but when it is formed, it can persist for very long periods. To date, there are very few cases with toxicological analyses performed on adipocere. We report two cases with a long delay between death and the discovery of the body (24 days and 12 years) associated with adipocere formation. In both cases a screening was carried out by GC-MS and LC-DAD. Additionally, a LC-MS/MS screening for drug facilitated sexual assault was performed allowing the detection of psychotropic substances at very low levels. Adipocere samples were hydrolyzed with a lipase buffer before liquid-liquid extraction. In the first case, diazepam, nordazepam and flunitrazepam were found at 2.1 ng/g, 1.8 ng/g and 1.2 ng/g, respectively. In case 2, phenobarbital was found at 26 μg/g in adipocere and 59 μg/g in liver. Although the post-mortem delay was important and the environmental conditions may have affected the measured concentrations, we were able to highlight the presence of 3 benzodiazepines and a barbiturate. The interpretation of these results remains complicated because of the lack of data in the literature.

Following positive opiate urine testing in employees working in risk positions in a transport company while these subjects claimed to consume nothing other than poppy seed sandwiches, a study was carried out to evaluate the opiates content of poppy seed baguettes and sandwiches as well as the urine, saliva and plasmatic concentrations observed after consumption of these products. Eleven people took part in this study, 6 ingested a whole poppy seed baguette and 9 ate a sandwich. Urinary concentrations for opioids were measured for 48 hours, saliva was analyzed for 10 hours, and plasma levels were measured up to 3.5 hours in one subject. Analyses were performed using liquid chromatography – tandem mass spectrometry. In order to highlight biomarkers of exposure to poppy seeds, urine samples were also analyzed in high resolution mass spectrometry with a metabolomic approach. The concentrations found in poppy seeds for morphine, codeine, thebaine and laudanosine are 530 mg/kg, 80 mg/kg, 300 mg/kg and 10 mg/kg, respectively. Urine concentrations observed after ingestion of the baguette were all positive until 18 h with maximal average concentrations observed at H4 around 2800 ng/mL for total morphine and 700 ng/mL for total codeine. After ingestion of a sandwich, all urines tested positive for morphine until 18 h, with a maximal average at H4 around 1800 ng/mL and 360 ng/mL for total codeine. Some were still positive after 48 h. In plasma, the morphine concentrations were between 18 and 19 ng/mL, corresponding to pharmacologically active concentrations. Noscapine and 6-acetylmorphine are biomarkers for heroin intake, while thebaine and for the first time here, 6-O-desmethylthebaine-glucuronide and O-desmethyl-laudanosine-glucuronide are identified as biomarkers for poppy seeds ingestion. This study showed the presence of morphine and codeine in very large quantities in some commercially available breads. These concentrations, far superior from the European Food Safety Authority's (EFSA) recommendations, can have major consequences particularly from a forensic point of view or in the context of occupational medicine, but also for the health of the most vulnerable populations. The detection of thebaine should be systematically performed when looking for opiates in blood, urine and saliva to facilitate interpretation of the results., Suite à des dépistages urinaires positifs en opiacés chez certains employés travaillant sur des postes à risque dans une société de transport alors que ces sujets prétendaient ne rien consommer d’autres que des sandwichs aux graines de pavot, une étude a été menée afin d’évaluer la teneur en opiacés des baguettes et sandwichs aux graines de pavot commercialisés dans certaines boulangeries ainsi que les concentrations urinaires, salivaires et plasmatiques observées après consommation de ces produits. Onze personnes ont participé à cette étude, parmi elles 6 ont ingéré une baguette entière aux graines de pavot, et 9 un sandwich avec ces graines provenant de 2 boulangeries situées à Paris et à Versailles. Les concentrations urinaires des différents opiacés ont été mesurées durant les 18 h qui ont suivi l’ingestion des baguettes et les 48 h suivant l’ingestion des sandwichs, une recherche salivaire a été effectuée durant 10 h et les concentrations plasmatiques ont été mesurées jusqu’à 3,5 h chez un des sujets après ingestion du sandwich. Les analyses ont été réalisées en chromatographie liquide couplée à la spectrométrie de masse en tandem. Afin de mettre en évidence d’éventuels biomarqueurs spécifiques d’exposition au graines de pavot et discriminant d’une prise d’héroïne, les échantillons d’urine ont également été analysés en spectrométrie de masse haute résolution avec une approche métabolomique. Les concentrations retrouvées dans les graines de pavot sont pour la morphine, la codéine, la thébaïne et la laudanosine de 530 mg/kg, 80 mg/kg, 300 mg/kg et 10 mg/kg, respectivement. Après ingestion d’une baguette, toutes les urines sont positives pendant au moins 18 heures, durée du recueil, avec des concentrations moyennes maximales à H4 atteignant pratiquement 2800 ng/mL pour la morphine totale et 700 ng/mL pour la codéine totale. Après ingestion d’un sandwich, les urines sont toutes positives en morphine totale jusqu’à 18 heures avec des concentrations moyennes maximales à H4 de l’ordre de 1800 ng/mL et 360 ng/mL pour la codéine totale. Certaines sont toujours positives 48 h après ingestion. Dans le plasma, les concentrations de morphine retrouvées sont comprises entre 18 et 19 ng/mL, soit des concentrations actives pharmacologiquement. La noscapine et la 6-acetylmorphine ne sont retrouvées qu’après prise d’héroïne, alors que la thébaïne et pour la première fois décrit ici, la 6-O-desmethylthébaïne-glucuronide et la O-desmethyl-laudanosine-glucuronide ne sont présents qu’après absorption de graines de pavot. Cette étude a montré la présence de morphine et de codéine en quantité très importante dans certains pains aux graines de pavot disponibles dans le commerce. Ces concentrations, bien supérieures aux recommandations de l’Autorité Européenne de Sécurité des aliments (EFSA), peuvent avoir des conséquences importantes dans un cadre médicolégal ou dans un cadre de médecine du travail, mais également présenter des risques pour la santé sur les populations les plus fragiles. La recherche de thébaine devrait être réalisée systématiquement lors de la recherche d’opiacés dans le sang, les urines et la salive afin de discriminer une prise de graines de pavot d’une prise d’héroïne.

Resume Objectif Determiner l’interet du dosage de naltrexone (NTX) dans les cheveux comme marqueur de l’observance du traitement de l’alcoolo-dependance. Methode Deux meches de cheveux châtains de 6 cm d’une patiente traitee par naltrexone (Revia®, 100 mg/j depuis plus de 4 ans) pour alcoolo-dependance et jugee compliante ont ete analysees. Apres decontamination et lavage, la premiere meche est coupee en 3 segments de 2 cm, puis broyee. Vingt milligrammes de cheveux sont extraits en milieu basique a l’aide des melanges hexane/acetate d’ethyle (1:1, v/v), puis chloroforme/isopropanol (4:1, v/v) en presence d’hydromorphone-d3 (HM) comme standard interne. Les deux phases organiques reunies sont evaporees et le residu repris par 80 μL de phase mobile. 10 μL sont alors analyses par CL-SM/SM. La deuxieme meche de cheveux a ete utilisee pour le dosage d’ethyglucuronide (EtG) afin de confronter les resultats obtenus. Resultats La methode est lineaire sur l’intervalle 2,5 a 1000 pg/mg (R2 > 0,999). L’exactitude varie de 96,6 a 104,1 % (n = 6) et la precision totale (intra- et inter jours) est inferieure a 6,3 % (n = 18). La limite de detection et de quantification sont de 1 et 2,5 pg/mg, respectivement. Les concentrations de naltrexone retrouvees a 1500, 1450, et 810 pg/mg dans les trois segments semblent en faveur d’une exposition reguliere a la naltrexone dans les 6 mois precedant le prelevement. Les concentrations en EtG dans les segments 1–3 cm et 4–6 cm sont de 20 et 25 pg/mg, inferieures au seuil fixe par la Society of Hair Testing (SOHT) de 30 pg/mg, ce qui suggere une consommation moderee et non excessive d’alcool durant cette periode, probablement en lien avec les proprietes pharmacologiques de la naltrexone sur la diminution de consommation chez le sujet alcoolo-dependant. L’analyse capillaire a par ailleurs permis de controler la compliance de la patiente a l’integralite des molecules qui lui sont prescrites. Conclusion A ce jour, aucune donnee dans la litterature ne fait etat des concentrations de naltrexone mesurees dans les cheveux, et de l’interet de son dosage pour le suivi de l’efficacite du traitement. Il s’agit donc du premier cas de dosage de naltrexone dans les cheveux au cours d’un traitement chronique de l’alcoolo-dependance. Ces resultats, valides sur un plus grand nombre de patients, pourraient etre utiles pour le suivi therapeutique des patients alcoolo-dependants sous naltrexone.

Objectifs Les CS sont le plus grand groupe de NPS consommes dans le monde avec plus de 179 molecules identifiees en Europe depuis 2017, et representent donc un defi majeur en matiere de detection et de surveillance. Neanmoins, la realite de leur consommation en France demeure incertaine [1] . Nous rapportons le cas d’une polyconsommation de 9 CS et demonterons leur incorporation dans les cheveux a travers les concentrations observees. Methode Un homme de 42 ans est hospitalise en psychiatrie en decembre 2019 pour dependance au cannabis (10 joints/j) et alcool (180 g/j) suivie depuis 2008. Le patient est consommateur occasionnel de cocaine (1 prise/mois), de codeine, et ponctuellement d’heroine. Il presente des antecedents de troubles de la personnalite et une tentative de suicide et decrit un sevrage non respecte depuis 5 mois, suite au deces de sa mere. L’examen clinique revele un craving important avec sueurs, tremblements et palpitations. Un traitement anxiolytique et hypnotique est initie. Une analyse capillaire sur une meche noire de 20 cm (dreadlocks) a ete demandee pour caracteriser l’usage de drogues. Trois segments de 20 mg (A : 0–2 cm ; B : 2–4 cm ; C : 4–6 cm) sont decontamines, extraits par hexane/acetate d’ethyle (1 :1, v/v) en presence de JWH-018-d11 et analyses par LC-MS/MS en mode gradient (formiate d’ammonium 2 mM, 0.1 % d’acide formique (A) et acetonitrile (B)) sur une colonne Hypersyl Gold PFP (1,9 μm, 100 × 2,1 mm, ThermoFisher) maintenue a 30 C°. La detection est en mode MRM apres ionisation positive. La methode dediee a la recherche de plus de 300 molecules d’interet en toxicologie a recemment ete enrichie par l’ajout de 16 CS [2] . Resultats Les limites de detection (LDD) et de quantification des CS varient de 0,3 a 8 pg/mg et de 1 a 25 pg/mg respectivement (CV ≤ 15 %) et la limite superieure de quantification a 1000 pg/mg permet d’atteindre une bonne linearite (r2 > 0,996). La justesse intra-journaliere et inter-journaliere (biais) et les precisions (CV) correspondantes sont respectivement ≤ 8,2 %, ≤ 14,3 %, ≤ 8,3 % et ≤ 14,9 %. Le rendement de recuperation varie de 61 a 94 % et l’effet de matrice de 31 a 110 %. Aucune interference endogene ni de contamination inter-echantillons n’ont ete observees. L’analyse capillaire confirme l’exposition du patient au cannabis (A/B/C, pg/mg : THC 700/1071/1763 ; CBD 1096/1533/1977 ; CBN 433/813/1389), a la cocaine (350/230/960) et ses metabolites (BZE, EME, norcocaine, AEME, cocaethylene), ainsi qu’a la MDMA (9/5/5) et la codeine (180/170/30). De plus, 9 CS non declares par le patient ont ete identifies et quantifies : JWH-019 (50/27/12), JWH-200 (20/13/5), JWH-250 (30/42/40), AM-2201 (32/10/3), BB-22 (10/10/10), 5F-PB-22 (64/14/10), 5F-APINACA (20/ Conclusion Malgre une prevalence des CS qui demeure faible en France par rapport aux cathinones, ce cas illustre la necessite de les rechercher systematiquement et montre l’interet majeur des cheveux pour l’etude des expositions aux NPS.

According to French law, the roadside testing for drugs of abuse (DOA) should be performed in oral fluid (OF) using an immunological screening kit. If the screening is positive, confirmation has to be done in OF collected by a special swab, called the FLOQSwab™ (FS). Unlike other sampling kits, this device was not designed to collect OF since it does not contain an elution buffer. An analytical method was developed for the simultaneous detection of 10 DOA under control in France: tetrahydrocannabinol (THC) at 1 ng/mL, and cocaine, benzoylecgonine (BZE), morphine, 6-monoacetylmorphine (6-MAM), amphetamine, methamphetamine, 3,4-methylenedioxy-N-ethylamphetamine (MDEA), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxy-N-methylamphetamine (MDMA) at 10 ng/mL. Samples were eluted using the Quantisal® buffer and extracted by liquid-liquid extraction for THC and by solid-phase extraction for the remaining analytes. Analyses were performed by ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). The validated method made it possible to detect the concentrations required by law and was successfully applied to samples from drivers who screened positive. The main limitations of this kit are the large variability of the collected OF volume and the poor stability of DOA in OF, requiring the use of a conservation buffer.

There are many differences between the hair from children and that of adult subjects, the hair being thinner, more porous with a different growth rate from the usual 1 cm/month observed in adults. In order to determine whether hair analysis could discriminate between chronic use and acute administration of a drug in children like in adults, we analyzed hair from 18 children aged between 1 day and 15 years in whom the administration of different drugs was known (single therapeutic administration or acute intoxication). A strand of hair was sampled within 1 to 45 days after treatment or intoxication. Analysis was conducted using LC/MS/MS. In the 10 youngest children, aged between 1 day and 29 months, the compounds administered in hospital or responsible for intoxication (lidocaine, ropivacaine, diazepam, midazolam, levetiracetam, morphine, ketamine, methadone, buprenorphine, THC, MDMA) were found in all segments of the hair independently of the time of sampling (1–45 days after ingestion). The concentrations detected were similar along the hair shaft, showing a radial diffusion and incorporation of the analytes in the hair of young children from the sebum. Concentrations could be very high when sampled shortly after administration (72 ng/mg for methadone, 75 ng/mg for MDMA after 3 days) and lower when sampling later (1.2 ng/mg for MDMA after 45 days). In these cases, hair analysis allowed to highlight the compounds responsible for intoxication even when they had disappeared from the blood or urine but should not be used to discriminate long-term exposure to a drug. In the eight remaining children aged from 34 months to 15 years, the drugs used in hospital (lidocaine, diazepam, morphine) or responsible for intoxication (THC, codeine, buprenorphine) were not found in any analyzed segments sampled 1 to 5 days after administration of the drugs, in agreement with the non-incorporation of the drugs from the sebum into the hair. For those children aged over 34 months, hair analysis allows to determine the chronic administration of a drug, like in adults.

Objectif Developper et valider une methode permettant de quantifier 7 neuroleptiques dans le plasma et le sang total en LC-MS/MS par micro-extraction liquide–liquide dispersee (MELLD). Cette methode a ensuite ete comparee a une methode d’extraction liquide–liquide (ELL) conventionnelle, puis appliquee au suivi therapeutique pharmacologique (STP) de l’aripiprazole, ainsi qu’a deux cas medicolegaux. Methodes La prise d’echantillon pour les deux methodes de pretraitement (DLLME vs LLE) est de 200 μL de plasma ou de sang total. La DLLME utilise respectivement le chloroforme (0,1 mL) et le methanol (0,2 mL) comme solvants extractif et dispersif, la LLE etant realisee a l’aide de 3 mL d’un melange acetate d’ethyle:dichloromethane:heptane (40:22:38). Les molecules incluses dans l’etude sont : l’aripiprazole, le flupentixol, l’haloperidol, le penfluridol, la pipamperone, la trifluoperazine et le zuclopenthixol. Resultats Une bonne linearite est obtenue pour les deux methodes entre 1 et 1000 ng/mL. La comparaison a mis en evidence une excellente correlation pour les sept composes evalues (r2 > 0,98). L’effet matrice est moins important avec la DLLME compare a celui de la LLE : −24,1 a 7,5 % vs −59,3 a −39,7 % respectivement, ce qui a permis d’ameliorer considerablement la sensibilite de la methode malgre un rendement d’extraction de la DLLME nettement inferieur a celui de la LLE (15,8 a 23,9 % vs 60,6 a 107,7 %). Trente-cinq dosages ont ete realises dans le cadre du STP de l’aripiprazole. La mediane des concentrations plasmatiques etait de 148 ng/mL avec un intervalle interquartile allant de 64 a 260 ng/mL. Cette methode a egalement ete appliquee a deux cas medicolegaux. Cas no 1 : un homme de 26 ans aux antecedents de depression et de psychose est decede suite a un accident de voiture, l’analyse du sang peripherique a retrouve une concentration en aripiprazole a 7680 ng/mL. Cas no 2 : Une femme de 45 ans est retrouvee morte a son domicile, l’analyse du sang peripherique retrouve la presence de deux neuroleptiques, aripiprazole : 434 ng/mL et zuclopenthixol : 120 ng/mL, associee a 3 psychotropes, alprazolam : 130 ng/mL (hydroxy-alprazolam : 12 ng/mL), mirtazapine : 816 ng/mL (desmethyl-mirtazapine : 790 ng/mL) et acide valproique : 311 mg/L. Conclusion Nous avons developpe et valide une methode de MELLD sensible et rapide, utilisant un faible volume de solvant et permettant la quantification de 7 neuroleptiques dans le plasma et le sang total. Celle-ci a ete appliquee avec succes au STP de l’aripiprazole et a l’analyse de prelevements realises en post-mortem.

Objectif La consommation de drogue semble se generaliser parmi les hommes ayant des relations sexuelles avec les hommes (HSH) notamment lors de « chemsex ». Cette pratique serait associee a des comportements a risque plus importants. Toutes les etudes reposent toutefois sur le declaratif des HSH. Cette etude a pour objectif de mesurer la proportion de consommation de drogues dans cette population par analyse capillaire et son association aux pratiques a risque. Methodes Dans le cadre de l’etude ANRS Ipergay (evaluant la PrEP, prophylaxie pre-exposition contre le VIH), il a ete preleve chez des participants a une sous-etude 1 meche de cheveux tous les 4 mois afin de determiner leur consommation de substances (excepte GHB/GBL, poppers et THC). Une analyse segmentaire a ete pratiquee lorsque la longueur de la meche le permettait (≥ 3 cm). Apres decontamination et extraction liquide/liquide, un criblage en LC-MS/MS Haute Resolution Q-Exactive et une quantification par triple quadripole TSQ Vantage (Thermofisher®) ont ete effectues, incluant les stupefiants classiques (opiaces, cocaine, amphetamines, ketamine) mais egalement les nouveaux produits de synthese (NPS). Resultats Soixante-neuf volontaires ont ete inclus avec des caracteristiques demographiques similaires aux 429 participants de l’etude Ipergay. L’âge median est de 35 ans [28;41]. Deux cent dix-neuf segments de cheveux (1,5 a 2,5 cm) provenant de 137 meches ont ete analyses correspondant chacun a une consommation entre 1,5 et 2,5 mois. Trente-deux molecules ont ete identifiees, 9 considerees comme therapeutiques bien que probablement utilisees comme drogues recreatives (sildenafil, vardenafil, tadalafil). Huit stupefiants classiques ou substances detournees ont ete detectes sur au moins 1 segment chez 53 volontaires, soit une prevalence de 77 % : cocaine (n = 47, 68 %), MDMA (31, 45 %), ketamine (26, 38 %), metamphetamine (6), amphetamine (4), dextrometorphane (4), methylphenidate et butorphanol (1). Quinze NPS ont ete mis en evidence chez 27 patients, correspondant a une prevalence de 39 %. Par ordre de frequence, on retrouve la mephedrone (14), 4-MEC (11), ethylphenidate (5), methylone (4), methoxetamine (4), methiopropamine (3), PMMA (3), MDPV (3), metamfepramone (1), 5F-PB22 (1), diphenidine (1), phendimetrazine (1), phentermine (1), N-methyl-2-AI (1) et dimethylone (1). Aucune piperazine type TFMPP ou m-CPP n’a ete retrouvee. Aucun cas de consommation de NPS seul n’a ete retrouve : les NPS sont associes a la cocaine dans 25/27 cas (93 %), a la MDMA dans 20/27 (74 %) ou la ketamine dans 19/27 (70 %). L’analyse capillaire permet une meilleure detection des consommateurs que les auto-questionnaires puisque 21/53 (40 %) ayant des drogues dans les cheveux avaient declare ne pas en consommer. Les HSH consommant des drogues ont un nombre declare de partenaires dans les 2 mois precedant le prelevement significativement plus eleve que les non consommateurs (mediane = 7 [4;15] vs 5 [2;10], p Conclusion La prevalence de consommation des NPS et en particulier des cathinones chez les HSH utilisant la PrEP est elevee, mais moindre que celle des drogues classiques comme la cocaine et la MDMA. L’analyse capillaire est la seule methode qui permet d’evaluer precisement cette consommation, car sous-declaree. Cela est d’autant plus important que cette consommation s’accompagne d’une augmentation des pratiques a risque, en particulier du nombre de partenaires.

Background The aim of the present study is to describe the prevalence of NPS and conventional DOA in Paris and its suburbs over a six-year period using hair testing approach. Method Hair was sampled in patients admitted to different departments of Paris hospitals between 2012 and 2017. Two high-risk populations were mainly considered: 1) drug-dependent and 2) acutely intoxicated patients. Segmental hair analysis was performed by validated LC–MS/MS method to screen for DOA and 83 NPS. Results 480 patients (280 M/200 F, 15–70 years) were included. 141 patients tested positive for NPS (99 M/42 F; median age: 33). NPS prevalence was 29%, that of amphetamines, cocaine and opioids were 32%, 38.5% and 52%, respectively. 27 NPS were identified, 4-MEC and mephedrone (number of cases n = 24 each) were the most detected cathinones. JWH-122 (n = 1) was the only detected synthetic cannabinoid while ketamine (n = 104) was present in numerous NPS users (67%). 3-fluorofentanyl (n = 1), furanylfentanyl (n = 1), N-ethylpentylone (n = 2), pentedrone (n = 2), mexedrone (n = 1), methcathinone (n = 3), 6-APDB (n = 2), TFMPP (n = 2), 2-CE (n = 1), 3,4-MD-αPHP (n = 1) and dextromethorphan (n = 27) were identified for the first time in hair. Users were found to have more than one NPS in 53% of cases, mostly in combination with conventional DOA. The number of detected NPS rose from 5 in 2012 to 42 in 2017. A broad range of hair concentrations (0.001–318 ng/mg) was found, but the low median concentrations seem to show an occasional exposure more than chronic use. Conclusion NPS screening should be assessed in routine clinical practice, especially in high-risk populations.

We developed and validated a method to detect and quantify 12 anabolic steroids in blood (androstenedione, dihydrotestosterone, boldenone, epitestosterone, mesterolone, methandienone, nandrolone, stanozolol, norandrostenedione, tamoxifene, testosterone, trenbolone) and eight more in hair samples (nandrolone phenylpropionate, nandrolone decanoate, testosterone propionate, testosterone benzoate, testosterone cypionate, testosterone decanoate, testosterone phenylpropionate, testosterone undecanoate) using liquid chromatography coupled to high-resolution mass spectrometry. This method used a benchtop Orbitrap mass spectrometer operating with an APCI probe under positive ionization mode. Analysis was realized in full scan experiment with a nominal resolving power of 140,000. After addition of the internal standard (testosterone-D3) and incubation in phosphate buffer pH = 5 for hair, 200 μL of blood and 30 mg of hair samples were extracted with heptane. LOQ and LOD were determined at 5 and 1 ng mL−1 in whole blood and 10 to 100 pg mg−1 and 2 to 20 pg mg−1 in hair according to the compounds, respectively. The method was linear in the 5–1000 ng mL−1 range in whole blood and between 10 or 100 pg mg−1 and 1000 pg mg−1 in hair with correlation coefficients >0.99, and intra- and inter-day accuracy and precision were

4-Methylethcathinone (4-MEC) and 3,4-methylenedioxypyrovalerone (MDPV) are synthetic cathinones. The objective of this study was to develop a method in order to measure these compounds in hair of a patient. After decontamination, 20mg of hair were grinded and incubated in phosphate buffer pH 5.0 in presence of 100ng of MDMA-d5 used as internal standard. Double basic liquid-liquid extraction was performed. Samples were separated on a 1.9μm Hypersil GOLD PFP column (100×2.1mm) using gradient elution. Compounds were detected by a LCQ TSQ Vantage XP triple-quadrupole mass spectrometer. SRM transitions m/z 192.1→146.1 and 174.2, m/z 276.1→175.0 and 205.1 and m/z 199.1→165.1 were used for 4-MEC, MDPV and IS, respectively. The assay was accurate and precise over the range 0.001 (lower limit of quantification) to 1ng/mg in hair. No matrix effect was observed. The method has been applied to a 30-year-old man who usually consumed cathinones for 6 months administered intravenously and was admitted to a general hospital for delirious and tachycardia after absorption of 10g of a powder sold as 4-MEC and 5g of MDPV. Both 4-MEC (30ng/mg) and MDPV (1ng/mg) were identified in the hair at high concentrations showing a regular consumption of these drugs. Many others compounds were also identified (mephedrone, MDMA, MDA, cocaine and metabolites, tramadol, hydroxyzine, aripiprazole, haloperidol). Few data are available on concentration of these new designer drugs in hair however important in order to determine the acute or chronic consumption of these drugs.