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• Antidiabetic intoxications can be fatal. • Hair analysis is important to analyze the historical consumption of antidiabetics in cas of factitious hypoglycaemias. • A method for identifying and quantifying antidiabetic agents in hair is needed. • In the method developed by the authors, it is possible to simultaneously identify and quantify 13 different antidiabetics. Oral antidiabetics are the drugs used to control blood sugar in diabetic subjects. The greatest risk of using these drugs is hypoglycaemia, which can be fatal if managed inappropriately. The diagnosis of hypoglycemia may be simple in diabetic subjects but can become a challenge in subjects with no history of exposure to these drugs. The major interest of testing for these compounds in hair is in the case of unexpected hypoglycaemias, as it enables discrimination between hypoglycaemias caused by antidiabetics and other reasons (e.g. insulinoma). Therefore it is important for a toxicology laboratory to screen for antidiabetics in hair due to the large window of detection this matrix allows associated to its long stability over time. In this study, a method has been developed and validated using liquid-chromatography coupled to tandem mass spectrometry for the analysis of 13 oral antidiabetics in hair. After addition of three different internal standards (hydroxy-tolbutamide-d9 for sulfonylureas, repaglinide-ethyl-d5 for glinides and vildagliptin-d3 for gliptins) and incubation in an ultrasonic bath in methanol, the hair was dissolved in NaOH and then subjected to liquid–liquid extraction. The validation procedure demonstrated an acceptable linearity for all compounds between 1 and 50,000 pg/mg. LOD and LOQ were between 0.5 and 5 pg/mg and 1–10 pg/mg respectively. Repeatability and reproducibility were below 20 % at two concentrations for all the analytes. The method was successfully applied to the hair of 18 diabetic patients under treatment of oral antidiabetics. The hair tested positive for gliclazide (3–21,400 pg/mg), sitagliptin (1.4–1.8 pg/mg), vildagliptin (3.3 – 1,740 pg/mg) and repaglinide (14.1 pg/mg). [ABSTRACT FROM AUTHOR]

• Development and validation of a rapid method for SARMs in hair using UHPLC-MS/MS. • Simultaneous analysis of 9 SARMs and 2 metabolic modulators in hair. • First identification of S23 in authentic human hair in a doping case. Selective Androgen Receptors Modulators (SARMs) are a new class of doping drugs that emerged in sport since 2008. Easy access on the Internet also leads to their misuse by amateurs. It seems important for a laboratory of toxicology to develop a targeted screening of SARMs, given their health risks. A method has been developed and validated for the analysis in hair of 9 SARMs (AC262536, ACP-105, andarine, LGD-4033, MK-0773, MK 677, ostarine, RAD 410 and S23) and 2 other metabolic modulators (GW501516, SR9009), using liquid chromatography coupled to tandem mass spectrometry. After addition of bicalutamide-D 4 used as internal standard and incubation in phosphate buffer pH = 9.5, 20 mg of hair samples were extracted with liquid/liquid extraction. Linearity was verified for all compounds between 0.5 and 50 and 2000 pg/mg. LOD and LOQ were determined between 0.1–20 and 0.5–50 pg/mg respectively, according to the various analytes. Intra- and inter-day precision (CV < 20%), matrix effects and recovery were evaluated for all compounds with CVs < 20%. The application and the interest of SARMs screening was demonstrated in a doping case. Three SARMs were detected namely andarine (120–1644 pg/mg), ostarine (1–9 pg/mg) and S23 (0.6–16 pg/mg) in 6x1 cm segments of the subject. [ABSTRACT FROM AUTHOR]

During the Belle Epoque, the use of cocaine, well known to the South American populations, spread among the European elite in the form of a tonic drink created from coca leaves macerated in Bordeaux wine, the Mariani wine. Nowadays, bottles in circulation are extremely rare but the investigations of a Parisian historian led him to the discovery of a bottle of wine that belonged to Angelo Mariani's estate at the end of the 19th century. Our laboratory was asked to analyse the contents of the bottle in order to search for the main active compounds and to estimate the state of preservation. The analysis of the centenarian product collected was aimed at identifying the alcohols and solvents by HS-GC/FID and HS-GC/MS after dilution in water, and the main alkaloids contained in all the samples by LC-HRMS screening and LC-MS/MS quantification. The Mariani wine presented an alcohol content of 15.7°. The screening and the subsequent dosage revealed the following substances: cocaine (86 ng/mL), benzoylecgonine (383 ng/mL), cocaethylene (17 ng/mL), ecgonine methyl ester (130 ng/mL), caffeine, cuscohygrine, cinnamoylcocaine and synephrine. The alkaloids found in the Mariani wine confirm the real presence of coca leaves in this drink. Their low concentrations may indicate drug instability. The presence of cocaethylene demonstrates that it can be formed without passing through the liver, which was supposed to be the site of production when cocaine and ethanol are present in the body at the same time. These appear to be the first report in the literature presenting the analysis of Mariani wine with the dosage of cocaine derivatives. • First analysis of a 19th century bottle of Mariani wine using MS technologies. • Determination of the real presence of cocaine and metabolites in Mariani wine. • Evidence of the presence of cocaethylene formed outside the liver system. [ABSTRACT FROM AUTHOR]

• Protonitazene is a novel synthetic opioid, which belongs to the nitazene family. • Nitazenes have re-emerged on the new psychoactive substances market and have been reported in several fatal intoxication cases. • Protonitazene metabolism was studied in vitro after drug incubation with human liver microsomes. • The identified metabolites were compared to the responses obtained in two authentic human urine specimens.. Protonitazene, or N,N-diethyl-5-nitro-2-[(4-propoxyphenyl)methyl]-1H-benzimidazole-1-ethanamine, is a novel synthetic opioid, which belongs to the nitazene family. Over the last four years, nitazenes have re-emerged on the new psychoactive substances market and have been reported in several fatal intoxication cases. The metabolism of several nitazene analogues have already been studied, but to date, no data exists regarding protonitazene. The aim of the study was the detection of protonitazene and its metabolites in authentic human urine collected in two fatal intoxication cases, comparing the data after in vitro incubation with human liver microsomes, and subsequent analysis by ultra-performance liquid chromatography-tandem mass spectrometry and ultra-performance liquid chromatography-high-resolution mass spectrometry. Protonitazene metabolites, including N-desethyl-protonitazene, 5-amino-protonitazene and 4-hydroxy-nitazene, were characterized in vitro and were identified in the urine of both cases. The ratios between metabolites and parent protonitazene, higher than 1, were calculated to estimate the proportionality of metabolites. The results suggest that testing protonitazene metabolites should increase the window detection of exposure to protonitazene. [ABSTRACT FROM AUTHOR]

The selective androgen receptor modulators are a recent class of anabolic agents, used to improve athletic performance. Among these molecules, there is (2 S)-N-(4-cyano-3-trifluoromethylphenyl)− 3-(3-fluoro-4-chlorophenoxyl)2-hydroxy-2-methyl-propanamide, commonly known as S-23. This molecule appeared very recently on the doping market. As a result, very few data are available in the literature, and nothing has been published about long-term effects of S-23. The authors focused on the detection of S-23 and its metabolites in human urine, following a single oral administration of approx. 8 mg to a volunteer, using standard ultra-performance liquid chromatography-triple quadrupole-mass spectrometry (UPLC-MS/MS), and ultra-performance liquid chromatography-quadrupole time of flight-mass spectrometry (UPLC-Q-TOF-MS). To the best of the authors knowledge, this seems to be the first study ever achieved on S-23. In vitro experiment was performed, using human liver microsomes, in order to investigate the potential CYP- and UGT-dependent S-23 metabolites. Four metabolites were produced, which were identified as hydroxy-S-23 (C 18 H 12 O 4 N 2 ClF 4 : m/z [M-H-] 431.0423); O -dephenylate-S-23 (C 12 H 10 O 3 N 2 F 3 : m/z [M-H-] 287.0647); S-23-glucuronide (C 24 H 20 O 9 N 2 ClF 4 : m/z [M-H-] 591.0794) and hydroxy-S-23-glucuronide (C 24 H 20 O 10 N 2 ClF 4 : m/z [M-H-] 607.0743). After consumption of S-23, the parent drug was detectable in hydrolyzed urine from 2 h post administration up to 28 days, with concentrations ranging between 0.5 and 93 ng/mL. In the urine, only one of the four metabolites identified in vitro was detected, hydroxy-S-23. This metabolite was detected up to 28 days. It does not seem to increase the window of detection of S-23 as the ratio between hydroxy-S-23 and the parent drug was always lower than 1. Another metabolite, dihydroxy-S-23, not identified in vitro, was identified in the urine of the volunteer. Hair sample, collected one month after the consumption of a single tablet, was negative for S-23 and hydroxy-S-23, with a LOQ at 0.1 pg/mg. [Display omitted] • The authors focused on the detection of S-23 and its metabolites in human urine, following a oral administration of 8 mg. • Four metabolites were produced in vitro, using human liver microsomes. • Parent S-23 was detectable in hydrolyzed urine up to 28 days. • Hydroxy-S-23 was detected up to 28 days, in urine. Dihydroxy-S-23 was detected from + 8 hours up to 5 days. • Hair sample, collected one month after the exposure, was negative for S-23 and metabolites, with a LOQ at 0.1 pg/mg. [ABSTRACT FROM AUTHOR]

Clomiphene is a selective estrogen receptor modulator. It is indicated for the treatment of female infertility issues but in sport, it can be misused to stimulate endogenous testosterone secretion in men. Therefore, it has been prohibited at all times by the World Anti-doping Agency. The aim of this study was to get data to be able to interpret concentrations in athletes. A healthy volunteer (male, 62 years-old) ingested a single therapeutic dose of clomiphene (Clomid™, 50 mg). Strands of hair (blond, 4 cm) were collected one month after the ingestion. Body hair (beard, axillary, pubic and chest hair), and finger and toenails were collected over 4–5 months. A previous method was modified to identify and quantify clomiphene in keratinous matrices. 30 mg of specimen were sonicated and incubated in 1 mL of methanol, in presence of 200 pg of clomiphene-D 5 (internal standard). After centrifugation and evaporation of the organic phase, the samples were analyzed using LC-MS/MS. Linearity was verified in hair and nail clippings between 1 and 500 pg/mg. The limits of detection and quantification were determined at 0.3 and 1 pg/mg respectively. The study demonstrated that clomiphene tested positive in all the analyzed specimens at 9 pg/mg in head hair, from 28 to 486 pg/mg (body hair) and from 4 to 57 pg/mg (nails). Clomiphene was identified for the first time in multiple keratinous matrices. This study demonstrated that a single oral therapeutic dose is detectable in keratinous matrices over a long period of time. • A single dose of clomiphene (50 mg) is detectable in head hair, body hair and nail clippings. • A single therapeutic dose is detectable for a long period in keratinous matrices. • Higher concentrations of clomiphene are identified in body hair than in head hair. [ABSTRACT FROM AUTHOR]

Determining fetal death causes is a complex problem for the forensic pathologist. Beyond the medico-legal context, the expert must be able to evaluate the viability of the fetus at the time of death, to eliminate in-utero fetal death and to determine if the death is related to a fetal, a maternal, a placental cause, or simply related to obstetrical complications. The authors present the case of a 21-year-old woman who unexpectedly gave birth to a fetus during a party. As pregnancy was not acknowledged by the mother (regular menstrual cycles and use of hormonal contraception), no obstetrical check-up had been performed. She would have presented violent abdominal pain and expelled a mass in the toilet. The fetus body, enclosed in the amniotic pouch, and the placenta were found in the toilet. A forensic autopsy was performed jointly by a forensic pathologist and a specialist in fetal pathology. Histological, toxicological and genetic samples were collected. Body morphometry and bone maturation indicated a gestational age of 31–32 weeks of amenorrhea. A significant asphyxia syndrome and non-specific multi-visceral congestion were noted at autopsy. Histological analysis of the fetal tissues revealed a lung and skeletal muscle maturation in accordance with the estimated term. At the brain level, there were signs of anoxia and abnormal cortical development with periventricular nodular heterotopia areas. The placenta microscopic analysis revealed acute chorioamniotitis, the probable cause of the premature fetal expulsion. Toxicological analyses revealed the presence of ecstasy (48 ng/mL) and its metabolite MDA (2 ng/mL) in fetal blood. Although negative in blood, THC-COOH tested positive in urine (9 ng/mL). The fetus was repetitively exposed to cannabis, as Δ9-THC tested positive in hair (51 pg/mg). Maternal hair analysis on 4 × 3 cm evidenced a long-term use of cannabis, while results support single massive exposure to ecstasy. In this article, the authors try to explain the reflexive pathway carried out to establish death causes and the maternal toxic consumption imputability on the cerebral malformations and fetal death. This case illustrates both the interest of toxicological analyses in cases of fetal death and the importance of a collaborative work between forensic and fetal pathologists and toxicologists, which appeared critical to answer in the best conditions to the magistrates questions, as well as to the bereaved families. [ABSTRACT FROM AUTHOR]

A case of fatal left iliac penetrating wound in a thirty-year-old man, during a fight, was presented. The medical file described a transfixing five-centimeter large wound on the left iliac vein and a puncture wound on the left iliac artery. After performing an autopsy, we concluded that the death was secondary to multivisceral failure, secondary to hemorrhagic shock. The judge investigating the case requested a detailed forensic medical reconstructive opinion focusing on the sequence of events leading to the fatal blow. Our having access to the videorecording and the photographs of the crime scene, added to the testimonies of witnesses proved to be a great help to the investigation. The data from the crime scene video recordings allowed us to estimate the victim's physical activity after the fatal stabbing (running between 20 and 140 meters) and the time when he collapsed (between 32s and 1 min) after the stabbing which is rarely described with methods using Closed-Circuit Television material, especially in cases of fatal vascular injuries. Those data are usually estimated from eyewitnesses' accounts, which remain an unreliable method. • Using CTTV cameras allow documenting physical activity after fatal vascular injury. • In vascular injuries the vital prognosis depends largely on the venous lesions. • Further importance of the blood traces photography and vascular lesions description. [ABSTRACT FROM AUTHOR]