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Among the hundred cannabinoids present in Cannabis sativa indica , cannabidiol (CBD) is a phytocannabinoid discovered in 1940, which can account for up to 40% of the plant's extract. Medically, it has been proposed to treat convulsions, inflammation, anxiety and nausea. Contrary to the hallucinogenic ingredient of the plant, delta-9-tetrahydrocannabinol, CBD does not seem to have a sedative effect, which can increase its popularity among users. The identification of CBD in blood and urine has been widely described in the scientific literature for several years. Only few data have been reported regarding CBD identification in alternative specimens, such as oral fluid, sweat, exhaled breath and hair. CBD capsules were purchased in the USA from a grocery store and a green capsule containing 22 mg of CBD was orally administered to a 59-year-old healthy man. Oral fluid was collected over 8 h using the NeoSal™ device. Sweat was collected using PharmCheck™ sweat patch technology over 7 days. Exhaled breath was collected with the ExaBreath® DrugTrap device over 8 h. Beard hair was collected 7 and 14 days after administration. Finally, urine specimens were collected over 48 h in plastic tubes without preservative. CBD was only detected in oral fluid at 15 min, at 20 pg/mL. Increasing concentrations, up to 96 pg/patch of CBD, were detected in the sweat patches. CBD was detected for 45 minutes in exhaled breath (Cmax of 302 pg/filter at 30 min). CBD produced a very low but significant chromatographic signal in beard hair, with concentrations <1 pg/mg. Finally, CBD tested positive in urine after enzymatic hydrolysis with a Cmax at 70 ng/mL, after 6 h. [ABSTRACT FROM AUTHOR]

The objective of this publication is to present the interest of collecting several keratinous specimens in order to document possible drug impairment at the time of the assault, when knowledge solely occurred 7 months after. A subject committed a murder and within minutes after the crime self-inflicted serious wounds. He was charged to the hospital where he slowly recovered. After several weeks, he was sent to prison. During this period, intelligence indicated possible drug impairment at the time of the assault after using 25I-NBOMe and 4-MMC. Head hair (4 cm), axillary hair, and toenails were collected 7 months after the crime. New psychoactive substances were tested in each specimen using LC-MS/MS, which revealed the presence of 25I-NBOMe and 4-MMC in axillary hair (2 and 6 pg/mg) and toenails (1 and 5 pg/mg). However, the perpetrator claimed that the positive findings were due to contamination in prison. Therefore, the head hair was also tested and results returned negative (LOQ at 1 pg/mg), demonstrating absence of contamination during the last 4 months before collection. Combining the window of drug detection in axillary hair (about 4 to 8 months) and the one of toenail clippings (up to 8 months), and excluding drug exposure during the previous 4 months as well as external contamination as the head hair results were negative, allowed us to conclude that the positive findings in axillary hair and toenails are more likely than not consistent with consumption of both 25I-NBOMe and 4-MMC at the time of the crime. [ABSTRACT FROM AUTHOR]

Mirtazapine is an antidepressant drug, used to treat depression, but also, in some specific conditions, to treat obsessive–compulsive disorder and anxiety. Although mirtazapine is not a hypnotic, it can make the subject feel drowsy. Children under the age of 18 should not take mirtazapine, but for some very special diseases, a physician can prescribe it for a limited period of time. The authors report a case involving 2 children (7‐ and 9‐year‐old) who were administered mirtazapine without consent by the mother, who was under daily therapy with this antidepressant. Hair specimens, collected from the children were tested by liquid chromatography coupled to tandem mass spectrometry for mirtazapine and its metabolite, N‐desmethylmirtazapine, on 3 × 1 cm segments. The hair test results (3 × 1 cm segments) have demonstrated that both children have been repetitively exposed to mirtazapine for approximately the last 3 months before hair collection, with concentrations in the range 1.32–3.79 and 0.64–2.54 ng/mg for mirtazapine and N‐desmethylmirtazapine, respectively. Environmental contamination was ruled out as the measured concentrations are highly variable according to the pattern of drug distribution and the washes were negative. Hair testing for drugs appears as an excellent diagnostic tool for child protection toward drug exposure. [ABSTRACT FROM AUTHOR]

An athlete challenged the result from an in-competition doping test which returned with an adverse analytical finding for stanozolol, claiming it was due to supplement contamination. Her lawyer asked the laboratory to analyze several hair specimens simultaneously collected from five different anatomical regions, head, arm, leg, pubis and armpit, to document the pattern of drug exposure. A specific UPLC–MS-MS method was developed. After decontamination with dichloromethane, stanozolol was extracted from hair in the presence of stanozolol-d3 used as internal standard, under alkaline conditions, with diethyl ether. Linearity was observed for concentrations ranging from 5 pg/mg to 10 ng/mg. The method has been validated according to linearity, precision and matrix effect. Concentrations of stanozolol in head hair, pubic hair, arm hair, leg hair and axillary hair were 73, 454, 238, 244 and 7,100 pg/mg, respectively. The concentration of stanozolol in head hair is in accordance with data published in the literature. When comparing the concentrations, body hair concentrations were higher than the concentration found in head hair. These results are consistent with a better incorporation rate of stanozolol in body hair when compared to head hair. The simultaneous positive concentrations in different hair types confirm the adverse analytical finding in urine of the top athlete, as the measured concentrations do not support the theory of contamination. For the first time, an anabolic agent was simultaneously tested in hair collected from five different anatomical regions from the same subject, with a large distribution of concentrations, due to anatomical variations, and these findings will help interpretation in further doping cases when documented with hair. [ABSTRACT FROM AUTHOR]

• 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]

Insulin glargine is a long‐acting insulin analog that is converted after enzymatic cleavage of the arginine pair of the β‐chain into its main metabolite M1 (21A‐Gly‐insulin), which is responsible for the hypoglycemic activity. In all the overdose cases described in the literature, only M1 concentrations have been reported, whereas insulin glargine was always absent or below the limit of quantitation. In this study, we present a case of suicide of a young nurse by injection of insulin glargine in which the parent molecule was found at a toxic concentration in blood. The determination and the discrimination of insulin glargine from human insulin and other synthetic analogs in the blood specimen were performed by liquid chromatography coupled to high‐resolution mass spectrometry (Waters XEVO G2‐XS QToF) and extraction after precipitation in the presence of bovine insulin (internal standard), with a mixture of acetonitrile/methanol +1% formic acid followed by purification on solid phase extraction cartridges C18. Glargine insulin tested highly positive in the blood with a concentration of 1.06 mg/L. Due to the difficulty in obtaining a M1 pure standard, the metabolite could not be dosed. This unique presence of the parent molecule, reported for the first time, can be explained by inter‐individual variability in the rate of conversion to metabolite. Intravenous injection versus subcutaneous injection can also explain the presence of insulin glargine. Finally, the dose injected may have been so high that saturation of the proteolytic enzymes responsible for conversion to M1 should have occurred. [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]

Insulin aspart (NovoRapid®, NovoMix®, Novolog® and Fiasp®) is a fast-acting analog of human insulin, indicated in the treatment of type I and II diabetes. It is administered before meals to mimic the physiological insulin secretion that follows a rise in blood glucose. Its misuse for the purposes of suicide and murder and in the context of factitious order has often been described. In forensic medicine, the identification of insulin in biological samples has always been complex. In this paper, we present a case of suicide of a 64-year-old man who died after the injection of insulin aspart. He was suffering from terminal lung cancer and left a letter explaining the reasons for his suicide. Four empty NovoRapid® pens were found near the body. Body examination was unremarkable, and the femoral blood was collected in two dry Vacutainer™ tubes (red cap) and two sodium fluoride (NaF) tubes (gray cap). A liquid chromatography coupled to high-resolution mass spectrometry method was used to identify and discriminate insulin aspart from human insulin after immunopurification in the blood samples and in the pens. Blood specimens tested positive for insulin aspart with the concentrations of 36 and 37 ng/mL in dry tubes and 58 and 71 ng/mL in tubes containing NaF when tested ∼3 weeks after the collection of the specimens. The contents of the pens also matched with insulin aspart. The stability of insulin in blood is a critical point in the interpretation of the concentrations due to their rapid decrease caused by the activity of proteases in blood. During a degradation study implemented to compare three preservatives and dry tubes, suitable insulin aspart stability was observed with disodium salt of ethylenediaminetetracetic acid and NaF. Given that NaF is standard in forensic toxicology for measuring blood alcohol concentrations, the authors suggest its use for blood collection when insulin intoxication is suspected. [ABSTRACT FROM AUTHOR]

Abstract: Easily accessible biomarkers for fetal stress biology are lacking. We here explore whether quantification of major fetal steroids, dehydroepiandrosterone (DHEA) or DHEA sulfate (DHEAS), with liquid chromatography/tandem mass spectrometry in infant nails is a tool to assess fetal stress biology in response to maternal stressful life events during pregnancy. Sufficient nail (≥1mg) was available from 80 infants (93% of those providing samples). The concentration of DHEA, but not DHEAS, was increased in infants of mothers with stressful life events during pregnancy (DHEA: F 1,41 =6.105, P =0.018; DHEAS: F 1,77 =0.767, P =0.384). DHEA concentrations were not related to maternal stress before pregnancy (F 1,41 =0.010, P =0.922). Infant nail DHEA may be a fetal biological correlate of intrauterine exposure to maternal stress. The method promises the first non-invasive retrospective biomarker for intrauterine stress biology, opening new ways for research and clinical applications in fetal medicine, endocrinology, obstetrics, gynecology, and for understanding the developmental origins of health and disease. [Copyright &y& Elsevier]

Ricochet of a bullet in the spinal canal is well known by neurosurgeons but relatively not a common event in usual medico-legal autopsy practice. This article presents a homicide case of a penetrating gunshot injury of the lumbar spine through the T12-L1 intervertebral foramen with active movement of the projectile within the spinal canal to the L5-S1 level. This case illustrates a bullet intradural and intramedullary active movement because of a ricochet of the body of T12 with active redirection of the path. In the current literature, different types of migration in caudal or cranial direction, intradural, or intramedullary are reported. If spontaneous migration of T10 to S1 seems to be more frequent, some authors reported a C1 to S2 migration. Such migration could be asymptomatic or induce neurological impairment. The medico-legal consequences of these migrations within the spinal canal are described. [ABSTRACT FROM AUTHOR]

Abstract: Biomechanics of human head has been widely studied since several decades. At a mechanical level, the use of engineering allowed investigating injury mechanisms developing numerical models of adult head. For children, the problem is more difficult and evaluating child injury mechanisms using data obtained from scaling adult injury criteria does not account for differences in morphology and structure between adults and children. During growth, child head undergoes different modifications in morphology and structure. The present paper compares the anthropometry and numerical simulations of a child head model based on medical CT scans to a child head model developed by scaling an adult head model using the method proposed by Mertz [H.J. Mertz, A procedure for normalizing impact response data, SAE paper 840884, 1984]. These analysis point out significant differences showing that scaling down an adult head to obtain a child head does not appear relevant. Biofidelic and specific child geometry is needed to investigate child injury mechanisms. [Copyright &y& Elsevier]

Abstract: Objective:: Since the 1960s, glucocorticoids are used by athletes to improve their performances. Their use is restricted in sports. Hair can document chronic abuse and can be therefore a complementary matrix for doping control. We have developed a new extraction, purification, and separation technique using liquid chromatography and mass spectrometry for the identification and quantification of two endogenous glucocorticoids: cortisol and cortisone. Methods:: Qualitative and quantitative investigations were achieved with 44 hair samples (17 males, 27 females; age ranging from 2 to 90 years). Hair strands were washed in methylene chloride, the first two centimeters of the strand were cut and pulverized in a ball mill. The powdered hair was incubated in 2 mL Soerensen buffer, pH 7.6, for 16 h at 40°C, in the presence of cortisol-d3 as an internal standard. Purification of the incubation medium was achieved on SPE C18 Isolute extraction columns followed by an alkaline liquid–liquid extraction with diethylether. The eluate was evaporated to dryness and resuspended in 25 μL of acetonitrile/ammonium formiate (1:1,v/v). The chromatography was operated on a LC Packings Superba Nucleosil C18 column using a linear gradient of acetonitrile from 30% to 70% in 10 min. The detector was a Perkin Elmer Sciex API 100 mass spectrometer. The detector''s response was linear for cortisol and cortisone concentrations ranging from 1 to 500 pg/mg. Extraction recovery at 50 pg/mg was 74% for cortisol and 32% for cortisone. Repeatability (CV values n = 8) at 7 pg/mg cortisol and at 50 pg/mg cortisone were 11% in both cases. Limit of detection and limit of quantification were 1 and 5 pg/mg, for both compounds, respectively. Results:: Cortisol concentrations in hair ranged from 5 to 91 pg/mg (mean 18 pg/mg). Cortisone concentrations in hair ranged from 12 to 163 pg/mg (mean 70 pg/mg). No influence of hair colour could be found. Influence of sex on cortisone concentrations seems possible but could not be statistically demonstrated. Finally, cortisone concentrations in hair are significantly higher before the age of 20. Incorporation of cortisol and cortisone in hair could follow a passive diffusion through sweat after conversion of part of cortisol to cortisone by Type 2 11-β-Hydroxysteroid-dehydrogenase in sweat glands. This issue was documented by these analyses. [Copyright &y& Elsevier]

Promazine is one of the oldest phenothiazine derivatives that have been proposed for the treatment of various psychiatric disorders. The drug is available as tablets, as syrups and in injectable forms. Despite its prescription to millions of subjects, its detection in human hair has seldom been reported. The aim of the present work is to develop a specific method to identify promazine in human hair by liquid chromatography–tandem mass spectrometry and to apply it to a patient who was self-medicating. The method involves overnight incubation of 20 mg of cut hair in 1 mL of pH 9.5 borate buffer in the presence of amitriptyline-d3 at 40°C. The chromatographic separation was performed using a reverse phase column HSS C18 with a gradient elution for 15 min. Linearity was verified from 0.5 to 500 pg/mg (r 2 = 0.9996), after spiking blank hair with the corresponding amounts of promazine. The limit of detection was estimated at 0.1 pg/mg. The precision was lower than 20%. Promazine was detected in the hair of a psychotic subject at 228–270 pg/mg in a 3 × 1 cm segment. Given this was a patient who was self-medicating, her physician requested an immediate drug discontinuation. In a fresh hair specimen collected 3 months later, the proximal segment (0–1 cm) tested positive at 0.9 pg/mg, clearly indicating that the time to obtain a negative result after promazine discontinuation is about 3–4 months. [ABSTRACT FROM AUTHOR]

Evidence of an insulin overdose is very complicated in the medico-legal field. The analysis and subsequent interpretation of results is complex, especially when treating postmortem blood samples. The instability of insulin, the special pre-analytical conditions and the absence of specific analytical methods has led most laboratories not to analyze insulin in their routine with a consequent underestimation of cases. This paper aims to assess the difficulties associated with the analytical characterization of insulin by describing a case that typically represents most of the inconveniences encountered following a suspected insulin overdose. The case concerns a man found dead at home by his brother. After an external examination, which did not reveal a specific cause of death, toxicological analysis was requested which did not reveal any substance of toxicological interest. Only 9 months later, it was reported to the toxicologist that the subject was diabetic, on insulin lispro treatment and that three empty syringes were found next to his body. Following analysis by LC-high-resolution mass spectrometry, the presence of insulin lispro at a concentration of 1.1 ng/mL, a therapeutic concentration, was evidenced. Despite the low concentration found, overdose cannot be excluded and this paper will describe the criteria evaluated to reach this conclusion. This case highlights that the interpretation of a postmortem insulin concentration is very complex and requires the evaluation of various elements including the circumstances of death, the subject's medical history, the interval between death and sampling and the sample storage. [ABSTRACT FROM AUTHOR]

Investigates homicide by manual occipitoatlantal (OA) dislocation. Prevalence of malicious OA dislocation; Circumstances that is used as basis by coroners to establish OA dislocation with homicidal origin; Method used in performing postmortem examination when injury to the cranio-cervical junction is suspected.

Presents a case report of an attack using a Japanese samurai sword. Claims of the attacker and victim; Evidence discussion; Suitability of the sword for stabbing and cutting; Potential of stabbing wounds to give fatal injuries; Difference between stab and incise wounds.

Delta 9 tetrahydrocannabinol (THC), the main component of cannabis, has adverse effects on the cardiovascular system, but whether concomitant ethanol (EtOH) and aging modulate its toxicity is unknown. We investigated dose responses of THC and its vehicle, EtOH, on mitochondrial respiration and reactive oxygen production in both young and old rat cardiac mitochondria (12 and 90 weeks). THC dose-dependently impaired mitochondrial respiration in both groups, and such impairment was enhanced in aged rats (−97.5 ± 1.4% vs. −75.6 ± 4.0% at 2 × 10−5 M, and IC50: 0.7 ± 0.05 vs. 1.3 ± 0.1 × 10−5 M, p < 0.01, for old and young rats, respectively). The EtOH-induced decrease in mitochondrial respiration was greater in old rats (−50.1 ± 2.4% vs. −19.8 ± 4.4% at 0.9 × 10−5 M, p < 0.0001). Further, mitochondrial hydrogen peroxide (H2O2) production was enhanced in old rats after THC injection (+46.6 ± 5.3 vs. + 17.9 ± 7.8%, p < 0.01, at 2 × 10−5 M). In conclusion, the deleterious cardiac effects of THC were enhanced with concomitant EtOH, particularly in old cardiac mitochondria, showing greater mitochondrial respiration impairment and ROS production. These data improve our knowledge of the mechanisms potentially involved in cannabis toxicity, and likely support additional caution when THC is used by elderly people who consume alcohol. [ABSTRACT FROM AUTHOR]

Contexte et problématique: La formation des étudiants en médecine doit non seulement leur permettre d'acquérir de solides connaissances scientifiques, mais surtout de mobiliser ces connaissances efficacement pour résoudre les problèmes, dans le cadre de ce que l'on nomme le raisonnement clinique. Le raisonnement clinique désigne l'activité intellectuelle selon laquelle un soignant collecte des données dans l'environnement et les confronte à ses connaissances et expériences antérieures dans le but de les utiliser pour prendre une décision diagnostique et de prise en charge. Les temps de supervision clinique des étudiants par les cliniciensenseignants constituent des moments privilégiés pour soutenir et accompagner le développement de cette compétence chez les étudiants. Plusieurs stratégies ont été décrites dans la littérature scientifique pour aider les étudiants à développer leur raisonnement clinique. Nous avons choisi d'expérimenter la méthode SNAPPS avec des étudiants de cinquième et sixième année de médecine, en stage dans le service médecine légale de la faculté de médecine de Strasbourg. Cette approche, développée par Wolpaw et al. (2003) permet de révéler le raisonnement clinique des étudiants ainsi que leurs incertitudes, tout en respectant le temps limité dont dispose le clinicien-enseignant pour offrir une telle supervision. Description: Les auteurs présenteront les principes de la méthode SNAPPS, ses modalités de mise en oeuvre et les ajustements réalisés afin de la déployer dans un service de médecine légale. Cette activité innovante à la faculté de médecine de Strasbourg a été jugée très utile par les étudiants qui plébiscitent son utilisation dans d'autre stage clinique. Ils soulignent en particulier, au même titre que les cliniciensenseignants, son intérêt pour les aider à se poser les bonnes questions, pour prendre le temps d'y répondre par eux-mêmes lors d'une analyse bibliographique, et pour conforter leurs hypothèses grâce au feedback ciblé du tuteur. [ABSTRACT FROM AUTHOR]

Anabolic steroids are synthetic derivatives of testosterone, and their abuse can have numerous health consequences. Identification of this group of drugs has found applications in forensic toxicology, clinical situations, psychiatric disorders, and of course, anti‐doping violations. Although anabolic steroids are generally tested in urine and very occasionally in head hair, collectors can face the lack of standard specimens, and therefore, nail clippings can be the unique alternative choice. Although there is no possibility to perform segmental analyses using nail clippings, the window of drug detection is generally much longer in nail when compared to head hair (particularly in male subjects), that is, 3–8 months and 4–12 months for finger and toenail clippings, respectively. A new method was developed, including nail pulverization in a ball mill, sonication for 90 min in methanol, and a combination of liquid–liquid and solid‐phase extractions, followed by gas and liquid chromatography coupled to tandem mass spectrometry. To document the application of steroid testing in nail clippings, the authors present 6 authentic cases of abuse, involving stanozolol (7 and 24 pg/mg), nandrolone (6 pg/mg), trenbolone (26, 67, 81, and 89 pg/mg), drostanolone (8 and 11 pg/mg), and testosterone enanthate (14 pg/mg). Given concentrations were always in the low pg/mg range, the use of tandem mass spectrometry appears as a prerequisite. [ABSTRACT FROM AUTHOR]

The article offers the authors' insight on a Swedish study which was suggested to be irrelevant due to its impact in the justice system, front-line clinicians, and child protection workers. Authors mentioned that the study, which concluded that abusive head trauma (AHT) was impossible to diagnose, fails to consider AHT evidence, have lacked of important details, and fails in respecting fairness and transparency.

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]

The dead body of a 54-year-old man was found at home by his partner. He was off work due to depression. A letter with suicidal intention was present on the scene. He was known to be a heavy drinker, and near the body, an empty bottle of whisky was found. In addition, 2 empty blisters of Eliquis (apixaban) 5 mg, corresponding to 40 tablets, were identified. Apixaban is an oral anticoagulant, acting as a factor Xa inhibitor. Autopsy findings were mostly unremarkable, except numerous bruises and some superficial self-inflected wounds. Histology showed hematomas of calyces and renal pelvis and in the liver, several areas of perivenular haemorrhagic necrosis. Others organs were congestive. Femoral venous blood alcohol was 0.11 g/L. In femoral venous blood, a toxic concentration of apixaban was measured at 1184 ng/mL using LC-MS/MS. Other drugs found at therapeutic concentrations included diazepam (99 ng/mL), nordiazepam (171 ng/mL), flecainide (447 ng/mL), and mianserine (65 ng/mL). Using liquid chromatography coupled to high-resolution mass spectrometry, 2 metabolites were identified, O-desmethyl-apixaban (61.8% of the apixaban response) and hydroxyl-apixaban (4.5% of the apixaban response). Long-term therapy was confirmed by a concentration of 10390 pg/mg in pubic hair. [ABSTRACT FROM AUTHOR]

Testing for drugs in hair raises several difficulties. Among them is the interpretation of the final concentration(s). In a post‐mortem case, analyses revealed the presence of furosemide (12 ng/mL) in femoral blood, although it was not part of the victim's treatment. The prosecutor requested our laboratory to undertake an additional analysis in hair to obtain information about the use of furosemide. A specific method was therefore developed and validated to identify and quantify furosemide in hair by UHPLC‐MS/MS. After decontamination of 30 mg of hair, incubation in acidic condition, extraction with ethyl acetate, the samples were analyzed by UHPLC‐MS/MS. Furosemide was found in the victim's hair at 225 pg/mg. However, it was not possible to interpret this concentration due to the absence of data in the literature. Therefore, the authors performed a controlled study in two parts. In order to establish the basis of interpretation, several volunteers were tested (four after a single 20 mg administration and twenty‐four under daily treatment). The first part indicated that a single dose is not detectable in hair using our method. The second part demonstrated concentrations ranging from 5 to 1110 pg/mg with no correlation between dosage and hair concentrations. The decedent's hair result was interpreted as repeated exposures. In the case of furosemide analysis, hair can provide information about its presence but cannot give information about dosage or frequency of use. [ABSTRACT FROM AUTHOR]

A 23-month-old boy was brought to a medical center by his mother, as she noticed that the father has gripped him around the neck and this had left marks. As a result of this, a child protection medical examination was requested. However, there was a significant chronology of mental health issues in the mother. Among the mother's medications, quetiapine and propranolol were the more active. Given a consultant pediatrician was concerned that the boy was vulnerable and potentially has experienced neglect and physical harm, the local authority instructed a hair test to document possible poisoning. However, this occurred several months later, due to court delays (postponed hearings and decisions) when the child was 32-month old. The laboratory received a strand of hair of the child (12 cm in length, light brown in color) and a strand of hair of the mother (>20 cm in length, dark in color) with the request to test both specimens by segmentation (12 x 1 cm) for quetiapine, an anti-psychotic drug and propranolol, a β-blocker agent. After decontamination and segmentation, the specimens were incubated in borate buffer pH 9.5 and extracted by a mixture of ether/dichloromethane/hexane/isoamyl alcohol to test for the drugs, including norquetiapine by a specific LC–MS-MS method. The first 3 cm segments of the child's hair were free of drug, roughly corresponding to the period he was no more in contact with the mother. Propranolol tested positive in the other segments at 15–72 pg/mg, with a linear increase from the proximal to the distal end. This was also observed for quetiapine, with concentrations in the range 10–18 pg/mg. Norquetiapine was never identified in the child's hair. The following concentrations were observed in the mother's hair: 6028–10,284, 910–4576 and 1116–6956 pg/mg for propranolol, quetiapine and norquetiapine, respectively. This confirmed that the donor was a long-term repetitive user of propranolol and quetiapine. The hair test results have indicated that the child was in contact with propranolol and quetiapine for a long period. It is not possible to put a temporal period for each segment, as the hair growth at the age of 32 months is not the same as for an adult (difference in the duration of the anagen period), nor to put any quantitative dosage or frequency of exposure(s) when interpreting the data. An increase of concentrations from root to tip was observed which is considered highly indicative of external contamination, with the older hair segments (those which are the more concentrated) being in contact for a longer time with contaminated items (hands of the mother, home items such as furniture, dishes, beddings, etc.). Overinterpreting drug findings in hair can have very serious legal implications in child protection cases, particularly when no other toxicological test and no clinical report exist to support voluntary administration of drugs. Whatever the findings, a proper interpretation of hair test results is critical and should be done ideally with other information available, such as medical history, witness statements and the available circumstances of the matter. A single hair test should not be used to determine long-term exposure to a drug. [ABSTRACT FROM AUTHOR]

The retrospective calendar of an individual's drug use requires a multisectional analysis in which the length of hair, corresponding to the full temporal window available, is cut into shorter sections to measure drug use during shorter periods of time (generally 1 cm corresponds to ~1 month). Segmental hair analysis is used to verify both previous drug history and recent enforced abstinence. However, after drug discontinuation, the fresh new hair growth segment cannot be immediately negative, due to the contribution of dormant hair. The objective of the study was to test hair samples from chronic tramadol and cannabis users after the discontinuation of both drugs and to evaluate the delay to wait until the hair will become negative. Hair specimens were obtained from eight subjects with a known history of tramadol abuse. Hair was collected 3–6 months after tramadol discontinuation. Tramadol was tested by liquid chromatography coupled to tandem mass spectrometry (LC–MS-MS) with a LOQ at 5 pg/mg. A second set of hair specimens were obtained from 15 subjects with a known history of cannabis abuse. Hair was collected 6–9 months after cannabis discontinuation. THC-COOH was tested by LC–MS-MS with a LOQ at 0.2 pg/mg. The hair stands were cut into L × 1 cm segments, according to their length (L), and tested for the respective drug. It was asked to each subject to clearly indicate the date of drug discontinuation. Assuming a rate of hair growth of 1 cm/month, the segment corresponding to the time of last drug use was calculated. The older segment just before this one was considered as the 100% of the response. THC-COOH and tramadol concentrations in this segment ranged from 2.3 to 8.9 and 895 to 21,010 pg/mg, respectively. After cessation of drug consumption, the presence of both drugs in new growing hair segments continued for a certain period with a more or less broad transition zone. Negative hair results were obtained ~3–4 and 6–7 months after cessation of tramadol and cannabis abuse. [ABSTRACT FROM AUTHOR]

Clomiphene or clomifene is a selective estrogen receptor modulator used to treat female fertility in case of ovulatory dysfunction. In sport, clomiphene is prohibited at all times for use by athletes and is listed in the section S4.2 (hormone and metabolic modulators) by the World Anti-Doping Agency. Indeed, clomiphene can indirectly increase testosterone levels in the body and can mitigate some side effects of synthetic steroid abuse. Despite its prescription to millions of subjects, its detection in human hair or nail clippings has never been reported. The aim of this study was to develop a specific method to identify clomiphene in hair and nail clippings by liquid chromatography–quadrupole tandem mass spectrometry. The procedure was then applied in a case of challenged doping results. The method involves sonication/incubation for 1 h of 30 mg of pulverized material in 1 mL of methanol in the presence of 2 ng diazepam-d5 used as internal standard. The chromatographic separation was performed using a HSS C18 column with a 15 min gradient elution. After spiking blank hair and nail with the corresponding amounts of clomiphene, linearity was verified from 1 to 500 pg/mg (r 2 = 0.9994 and 0.9995 for hair and nail, respectively). The limit of detection was estimated at 0.3 pg/mg for both matrices. No interference was noted from endogenous compounds, particularly steroids. Clomiphene was identified at 85 and 20 pg/mg in the pubic hair and the fingernail clippings, respectively, of a male athlete challenging an adverse analytical finding. [ABSTRACT FROM AUTHOR]

Background: The detection and analysis of drugs in hair has progressively emerged as a consequence of the enhanced sensitivity of analytical techniques used in forensic toxicology; a greater advantage in using this matrix respect to classical ones (i.e., urine and blood) is an easier and noninvasive sample collection, even when the careful supervision of law-enforcement officers is required to avoid the risk that the sample may be adulterated or replaced. Moreover, according to the length of the hair, the history of drug exposure can be retrospectively monitored fromfew weeks up to months or years since sample collection. Objective: Given the potential negative effects of pregabalin, an antiepileptic and analgesic drug with a high risk of misuse and abuse, the laboratory was asked to test for the drug in hair. Method: A new ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry was developed. Themethod involves incubation of 25 mg of cut hair in acetonitrile for 2 h in an ultrasonic bath and separation on an Acquity HSS C18 column (150×2.1mm×1.8 µm) maintained at 50°C in a thermostatically controlled oven. A gradient elution was performed. Results: The method was fully validated according to international standards. The limit of quantitation of the test was 10 pg/mg. Five authentic cases of pregabalin in hair segmentswere tested using the method and the resultswere in the range 17-1487 pg/mg. Conclusion: This new method was found suitable to monitor both patients under pregabalin therapy and dependent subjects. [ABSTRACT FROM AUTHOR]

3-Hydroxyphencyclidine (3-OH-PCP) is a hydroxy derivative of phencyclidine, synthesized in 1978 to investigate the structure–activity relationship of phencyclidine derivates. In vitro studies have shown that 3-OH-PCP, like phencyclidine, acts on the N-methyl-D-aspartate receptor and has a higher affinity for this receptor than phencyclidine. The authors report the case of a 38-year-old man, known for drug addiction, found dead at home with two plastic bags of powders found near his body. Using liquid chromatography coupled to tandem mass spectrometry, peripheral blood toxicological analysis revealed consumption of 3-OH-PCP with a concentration of 3-OH-PCP being 524 ng/mL. Blood also tested positive for nordiazepam, methylphenidate, amisulpride, methadone and benzoylecgonine, all at concentrations near those observed after recreational abuse. The blood concentration of 3-OH-PCP is the highest ever reported in the literature. Hair testing also revealed 3-OH-PCP, at 174 pg/mg, which may correspond to a chronic consumption of this molecule. A nuclear magnetic resonance analysis of the two powders highlighted 3-OH-PCP and 5-methoxy-dimethyltryptamine, estimated to have a purity of 85.4 and 91.3%, respectively, using the Electronic Reference To access In vivo Concentrations method. [ABSTRACT FROM AUTHOR]

The article discusses the chemical stability of Δ9-tetrahydrocannabinol (THC), the active ingredient and its two major metabolites, 11-hydroxy-Δ9-tetrahydrocannabinol (11-OH-THC) and 11-nor-9-carboxy-Δ9-tetrahydrocannabinol acid (THC-COOH), in whole blood in presence of formalin solution. It mentions that formalin has a high destructive effect on cannabinoids, both parent and metabolites; and also mentions that formalin, concentrations of both metabolites of cannabis are lower than THC.

A 29-year-old man with no previous medical history was found dead at home. Anabolic products (tablets and oily solutions) and syringes were found at the scene. The man was known to train regularly at a fitness club and to use anabolic drugs. Following an unremarkable autopsy with normal histology, toxicological analyses were requested by the local prosecutor to provide further information. Blood, head hair (5 cm, black), body hair (axillary and leg) and toe and finger nail clippings were submitted to liquid and gas chromatography coupled to tandem mass spectrometry (LC and GC–MS-MS) methods to test for anabolic steroids. Blood tested positive for testosterone (4 ng/mL), boldenone (26 ng/mL), stanozolol (3 ng/mL) and trenbolone (<1 ng/mL). Segmental head hair tests (2 × 2.5 cm) revealed a repeated consumption of testosterone (65–72 pg/mg), testosterone propionate (930–691 pg/mg), testosterone isocaproate (79 pg/mg to <5 pg/mg), nandrolone decanoate (202–64 pg/mg), boldenone (16 pg/mg), stanozolol (575–670 pg/mg), trenbolone (4 pg/mg–not detected), drostanolone (112–30 pg/mg), drostanolone enanthate (26–5 pg/mg) and drostanolone propionate (15–4 pg/mg). In addition to the substances identified in head hair, testosterone decanoate, testosterone cypionate and nandrolone were identified in both body hair and nails. The experts concluded that the manner of death can be listed as toxic due to massive repetitive use of anabolic steroids during the previous months. For anabolic agents, blood does not seem to be the best matrix to document a fatal intoxication. Indeed, these products are toxics when abused long term and are known to cause cardiac, hepatic and renal diseases. When compared to blood, hair and nails have a much larger window of detection. Therefore, keratinous matrices seem to be the best approach to test for anabolic steroids when a sudden death is observed in the context of possible abuse of steroids. [ABSTRACT FROM AUTHOR]

Fingerprints are invisible traces that result from a deposition of sweat and sebum present on the papillary ridges. As sweat and sebum contain drugs, fingerprints are promising since collection is rapid, non-invasive and difficult to falsify. Very limited data are available in the literature, and therefore, it seems opportune to study the transfer of xenobiotics onto the items taken in hand via the fingerprints. Two studies were implemented using the ballpoint pen as a model. The objective of the first study was to compare the nicotine concentrations found on the pens of three smokers and three non-smokers. Five pens, belonging to each subject and used regularly, were rubbed with a cotton swab dipped in methanol and analyzed by liquid chromatography coupled with tandem mass spectrometry (LC–MS-MS). The second study was to analyze the transfer via fingerprints of four volunteers, after administration of 30 mg of codeine. The objective was to determine the feasibility of this study and the time corresponding to the highest concentration of codeine. Over a 24-h period, new pens were handled for 5 min by the four volunteers, rubbed with a cotton swab dipped in methanol, and then analyzed by LC–MS-MS. The nicotine study showed a major difference between the nicotine concentrations obtained from smokers (between 6 and 276 ng/pen) and non-smokers (between 2 and 4 ng/pen). After administration of 30 mg of codeine, the analysis of the pens of the four volunteers allowed to demonstrate the presence of codeine up to 24 h between 9 and 544 pg/pen. Normal hygiene practices did not influence the final result. The highest concentration was observed after 2 h. Morphine was also detected (between 19 and 33 pg/pen). These preliminary results should be considered a demonstration of the interest of fingerprints testing to document drug exposure. [ABSTRACT FROM AUTHOR]

The article discusses the impact of tramadol's single exposure and application to a munchausen's syndrome by Proxy Case. It mentions long-term prognosis for the children depends on the degree of damage created by the illness or impairment and the amount of time it takes to recognize and diagnose MBPS. It also presents a case study a 8-month old boy suffering from intense sedation and dyspnea is presented.

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]

A 60-year-old man was found dead at home by his wife. The man was known to be a regular trainer in a fitness center and was described as a long-term user of anabolic steroids. The autopsy revealed heart failure with cardiomegaly, visceral congestion, and pulmonary edema. The local prosecutor ordered toxicological analyses but no anatomic pathology test. Initial analyses by the liquid chromatography coupled to quadrupole time-of-flight mass spectrometry system (UPLC--q-TOF-MS) system indicated the presence in femoral blood of trenbolone, an anabolic steroid. Trenbolone was quantified with a liquid chromatography system coupled with a tandem mass spectrometer in various autopsy specimens, including hair. Concentrations of trenbolone were 3.9, 3.2, 231, and lower than 0.5 ng/mL in femoral blood, cardiac blood, bile, and vitreous humor, respectively. Moreover, in head hair, the concentration was 9 pg/mg. The presence of trenbolone in hair can be interpreted as repetitive exposures when compared to the limited literature data. Repetitive misuse of trenbolone can explain advanced heart failure. This paper highlights the importance of testing biological specimens for anabolic steroids in case of heart failure. [ABSTRACT FROM AUTHOR]

The identification and quantification of insulin and its analogs have always been a challenge in the forensic field. Murder, suicide attempts and induced hypoglycemia in the context of factitious disorders have been described with the use of synthetic analogs of human insulin. There is very less information in the literature about aspart insulin concentrations in overdose cases. In this paper, we present a case of a nurse who tried to murder her 10-year-old daughter by injecting her aspart insulin and who, later, tried to commit suicide by injecting herself the same hormone. Two empty syringes and a FIASP® Flextouch pen were found in the woman's apartment. A liquid chromatography coupled to high resolution mass spectrometry (LC–HRMS) method was developed in order to identify and discriminate aspart insulin from human insulin in blood samples as well as in syringes and pen, while an liquid chromatography coupled to tandem mass spectrometry (LC–MS-MS) method was developed for the quantification of insulin in blood samples. Aspart insulin tested positive at 5.7 and 2.4 ng/mL in the blood specimens of the mother and the child, respectively. The substance contained in the syringes and pen also corresponded to aspart insulin. Although the mother claims to have injected an overdose of aspart insulin, the concentrations found were in the therapeutic range for subjects under therapy. Due to the high instability of insulin and the long time elapsed between sampling and forensic analysis (8 months) due to administrative reasons, the concentration at the time of collection was probably much higher. In this case, it was possible to identify aspart insulin and discriminate it from human insulin in a context of attempted murder and subsequent attempted suicide using high-resolution mass spectrometry, which is of paramount importance in forensic medicine. [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]

Introduction: Fire death investigations attempt to determine whether a subject was alive or dead before the fire started. Therefore, it is essential to assess if the bone damage is traumatic or the result of exposure to heat. This observational study aims to expose the specific CT semiology of thermal bone lesions to allow the forensic radiologist to identify and distinguish them from traumatic lesions that would have preceded death. Materials and methods: We retrospectively reviewed the CT findings of 25 bodies with thermal bone lesions for which a postmortem computed tomography (PMCT) was performed prior to an autopsy. Imaging findings were correlated to the autopsy data to identify the specific features of thermal bone lesions. Results: Specific signs of thermal injuries to bone were identified on PMCT on all deceased cases. Thermal damages predominated in areas directly exposed to flames (rib cage, distal extremities) with less soft tissue coverage ("soft tissue shielding"). The mottled appearance of bone marrow was a constant sign of burned bones. Heat fractures such as trans-diploic fractures of flat bones and beveled ("flute-mouthpiece") fractures of extremities seemed specifically related to thermal mechanism. In addition, we provided a better description of superficial cortical fissures of flat bones ("ancient Chinese porcelain") and observed a "stair step" fracture of a long bone until now undescribed in radiological literature. Conclusion: Thermal bone lesions have specific CT findings, different on several points from traumatic injuries. Their knowledge is essential for radiologists and forensic physicians to provide an accurate report of injury and conclusions. [ABSTRACT FROM AUTHOR]

Coca tea is a popular drink in some countries of South America where it is reputed to have medicinal properties. This preparation is composed of natural cocaine (COC) alkaloids and therefore can be banned in some countries. During an anti-doping control in Peru the urine of an athlete tested positive for benzoylecgonine (BZE) ecgonine methyl ester (EME) and COC (400 180 and 0.5 ng/mL respectively). The athlete indicated that she had consumed coca tea in the morning before the competition. As her lawyer contacted us to assess the scientific aspects of the possible involvement of coca tea to explain the adverse analytical finding a study was implemented with similar tea bags urine specimens were collected for each subject for 3 days to follow the elimination of COC and metabolites (BZE and EME). All samples were analyzed byultra high performance liquid chromatography tandem mass spectrometry after alkaline extraction. Maximum detection times for COC was 20 h with concentrations ranging from 6 to 91 ng/mL. Maximum detection times for BZE and EME were 70 h and 60 h respectively with concentrations ranging from 6 to 3,730 ng/mL and from 6 to 1,738 ng/mL. The concentration profiles were identical for the five volunteers. This study supports the athlete's claims. In addition, the sample of hair strands of the five subjects was collected a month later and all the hair tests showed a negative result for COC with a limit of decision of 10 pg/mg. Although it is accepted that a 4 mg dose of COC has no significant pharmacological effect the consumption of coca tea can lead to significant legal consequences since the measured urine concentrations sometimes cannot be considered incidental. Therefore, discrimination between coca tea consumption and recreational COC abuse relies primarily on hair analysis. [ABSTRACT FROM AUTHOR]

A 59-year-old male was found dead at home, with two empty vials of an oily preparation obtained from a manufacturer from East Europe. There was no label on the vial. The subject was a former weightlifter, also known as an anabolic steroids abuser. The local prosecutor ordered a body examination, which was unremarkable, and allowed collecting femoral blood, urine and scalp hair (6 cm, brown). He was treated for cardiac insufficiency with quinidine. Biological specimens were submitted not only to standard toxicological analyses including a screening with liquid chromatography (LC)–quadrupole time of flight, but also to a specific LC–tandem mass spectrometry method for anabolic steroids testing. Ethanol was not found in both blood and urine. Quinidine blood concentration (791 ng/mL) was therapeutic. No drug of abuse was identified. In blood, testosterone was less that 1 ng/mL and no other steroid was identified. In urine, testosterone/epitestosterone was 1.56 and boldenone was present at a concentration of 9 ng/mL. The hair test results, performed on the whole length, demonstrated repetitive steroids abuse, including not only testosterone (140 pg/mg), testosterone propionate (605 pg/mg) and testosterone decanoate (249 pg/mg), but also boldenone (160 pg/mg), trenbolone (143 pg/mg) and metandienone (60 pg/mg). Since forensic laboratories have limited access to steroid urinary metabolite reference material due to specific regulations (to avoid testing athletes before anti-doping verifications), hair analyses seem to be the best approach to document anabolic agents abuse. Indeed, in hair, the target drug is the parent compound; in addition, when compared to blood or urine, this matrix has a much larger window of detection. The pathologist concluded cardiac insufficiency in a context involving repetitive abuse of anabolic drugs. This case indicates that more attention should be paid to anabolic steroids, in a context of sudden cardiac death. [ABSTRACT FROM AUTHOR]

• Letrozole can be diverted from its therapeutic use. • Letrozole counteracts feminizing effects due to anabolic steroids abuse. • Hair is the specimen of choice to document long-term drug exposure. • Two independent methods (LC-MS/MS and LC-HRMS) are essential for legal rules. Letrozole is a reversible aromatase inhibitor, used in the treatment of hormone-dependent woman cancer. No indication for medical use is available for men. In recent years, several cases of doping with letrozole have been observed, especially among high level athletes. Aromatase inhibitors reverse the harmful effects (feminizing) of the abuse of anabolic androgenic steroids. Letrozole is included on the list of products prohibited in- and out-competition by the World Anti-Doping Agency, under section S4.1. The aim of the present work was to develop a specific method to identify letrozole in human hair of a male amateur athlete by LC-MS/MS and confirmation by LC-HRMS, after incubation of 20 mg of matrix in 1 mL of methanol. The chromatographic separation was performed using a reverse phase column HSS C18 with a gradient elution of 15 min (from 87% to 5% of formate buffer adjusted to pH 3). Linearity was observed from 1 to 1000 pg/mg (r2 = 0.9999), after spiking blank hair with the corresponding amounts of letrozole. The limit of detection was estimated at 0.5 pg/mg and the lower limit of quantification was the first point of the calibration curve, i.e. 1 pg/mg. The precision was lower than 20% and there was no interference with the analytes by chemicals or any extractable endogenous materials present in hair. Letrozole was identified in the male amateur athlete hair at 310 pg/mg (segment 0–2 cm) and 245 pg/mg (segment 2–4 cm). [ABSTRACT FROM AUTHOR]

The discovery of exogenous particles in the broncho-pulmonary tree is frequently described in forensic literature, especially in lung samples, in the context of aspirated gastric content during the death agony period or during resuscitation. We report an original observation of a multi-visceral dispersion of exogenous particles detected, in an 8-year-old boy, who allegedly fell from a 2-m high brick-wall. The autopsy found major liver fracture and diaphragm rupture with massive internal hemorrhage without gastric wall rupture. The histological analyses have identified round to oval bodies in the lung bronchi, alveoli, and, rarely, in vascular sections, and also on the surface of several samples. These particles stained strongly pink by the periodic acid Schiff method, evoking dried vegetables. Two hypotheses were invoked: aspirated vegetable particles into the bronchial tree or parasitic infection, like pinworm larva. In order to characterize the nature of these particles, different legumes were cooked, embedded in paraffin wax, and examined under light microscope. Simultaneously, morphological comparison between the gastric content and pinworm larva and lentils was made and a PCR analysis was performed on gastric fluid sample. The DNA sequencing showed a Fabaceae plant family, Lens culinaris. The possibility of a hematogenous dissemination of the starch grains during a perimortem aspiration of gastric content seems unlikely, and a contamination from the gastric content of the organs samples during the autopsy or the pathologic macroscopic and microscopic processes seems to be the principal hypothesis. The formal identification of such particles is important to avoid the misdiagnosis of a potential parasitic infection. The risk of confusion can be detrimental in some circumstances. [ABSTRACT FROM AUTHOR]

Introduction: Chloramphenicol (2,2-dichloro-N-[1,3-dihydroxy-1-(4-nitrophenyl)porpan-2-yl]acetamide) is a bacteriostatic antibiotic of the phenicolated family, used in the past to treat meningitis, plague, cholera, or typhoid fever. Treatment with chloramphenicol can have life threatening side effects, the most serious of which is aplastic anemia, which may be fatal. For this reason, the antibiotic was removed from the French market in 2008. Case report: In this paper, the authors report the case of a woman consuming chloramphenicol possibly in the context of factitious disorder. After a capsule containing chloramphenicol was discovered in her hospital bed, a hair specimen (about 16 cm, brown, not oriented) was collected and sent to the toxicological laboratory in order to document exposure to chloramphenicol. Results: The drug was identified in the hair specimen of the subject at 13.7 ng/mg. Discussion: Identification of chloramphenicol in hair has not been reported in the literature. As consequence, the interpretation of the concentration, the dosage and the frequency of abuse are difficult to establish. Conclusion: Given the context, physicians considered the case as a possible factitious disorder, thus being a unique observation of using chloramphenicol in such a context. [ABSTRACT FROM AUTHOR]

Betel nut is the fruit of Areca palm, growing in Papua New Guinea. Mixed with limestone and stick mustard, arecoline and guvacoline, which are present in betel nut, are hydrolyzed into arecaidine and guvacine, respectively. As part of the study on dietary habits of Papuans residents, our laboratory was asked to analyze the four alkaloids in hair to document long-term exposure. Hair samples were collected from 19 adult subjects (males = 11; females = 8), by some of the authors, and were sent to the laboratory for analysis. The four alkaloids have very similar chemical structures. In order to accurately identify the drugs, two methods were developed. First, the compounds were identified using an ultra-high-performance liquid chromatography system coupled to time-of-flight mass spectrometry. Then, they were quantified by an ultra-high-performance liquid chromatography system coupled to tandem mass spectrometry. After decontamination with dichloromethane, hair samples were cut into very small segments and 20 mg were incubated in methanol for 2 h 30 min in an ultrasound bath. After cooling, the methanol was evaporated to dryness in presence of 20-μL octanol to prevent volatilization. Nicotine-d4 was used as an internal standard. Linearity was observed for concentrations ranging from the limit of quantification to 20 ng/mg for arecoline, arecaidine, guvacine and guvacoline. Measured concentrations were in the range 60 pg/mg to 18 ng/mg for arecoline (n  = 19), 14 pg/mg to 2.5 ng/mg for guvacoline (n  = 11), 63 pg/mg to 3.8 ng/mg for arecaidine (n  = 11) and 100 pg/mg to 3.2 ng/mg for guvacine (n  = 6). There was no correlation between concentrations of arecoline and arecaidine (ratio from 0.01 to 0.18) and guvacoline and guvacine (ratio from 0.06 to 3.50). However, the identification of these substances in hair is a good marker of consumption of betel nut and allows us to document a local practice that remains difficult to evaluate just by questioning. [ABSTRACT FROM AUTHOR]

The body of a 61-year-old man was found at his home by his wife, lying on the floor, near the bathroom, around midnight. He was known to be training for bodybuilding, using anabolic steroids. Police investigations revealed the presence of two types of tablets at home, one supposed to contain clenbuterol (0.040 mg) and the other stanozolol (10 mg). Testing the tablets revealed different dosages from what was expected, i.e. 0.073 and 11.5 mg/tablet, for clenbuterol and stanozolol, respectively. External body examination and autopsy, which was performed the next day, revealed generalized organ congestion and lack of any traumatic injury (confirmed by radiology). Cardiomegaly, with a heart weighing 692 g, was obvious. Anatomic pathology tests did not reveal evidence of malformations, but atheromatous plaque was identified in the coronaries during complete histology investigations. Femoral blood, urine, bile, gastric contents and two strands of hair (6 cm) were collected for toxicology. These specimens were submitted to standard analyses, but also to a specific LC–MS-MS method for clenbuterol and stanozolol testing. Clenbuterol was identified in all the tissues, including femoral blood (1.1 ng/mL), urine (7.2 ng/mL), bile (2.4 ng/mL), gastric content (3.2 ng/mL) and hair (23 pg/mg). Stanozolol only tested positive in hair (11 pg/mg). All other analyses were negative, including blood alcohol and drugs of abuse. The pathologists concluded to cardiac insufficiency with support of cardiomegaly, in a context involving repetitive abuse of anabolic drugs. This case indicates that more attention should be paid to clenbuterol, a drug widely used as a stimulant by people who want to lose weight, athletes and bodybuilding practitioners. [ABSTRACT FROM AUTHOR]

• Girls are better able to delay gratification than boys. • Boys with higher prenatal testosterone levels are less able to delay gratification. • Boys with higher prenatal testosterone levels show more attentions problems/overactive behavior. • Boys with more attention problems/overactive behavior are less able to delay gratification. Sex differences in self-control become apparent during preschool years. Girls are better able to delay their gratification and show less attention problems and overactive behavior than boys. In this context, organizational effects of gonadal steroids affecting the neural circuitry underlying self-control could be responsible for these early sex differences. In the present study testosterone levels measured in amniotic fluid (via ultra performance liquid chromatography and tandem mass spectrometry) were used to examine the role of organizational sex hormones on self-control. One hundred and twenty-two 40-month-old children participated in a delay of gratification task (DoG task) and their parents reported on their attention problems and overactive behavior. Girls waited significantly longer for their preferred reward than boys, and significantly more girls than boys waited the maximum period of time, providing evidence for sex differences in delay of gratification. Boys that were rated as suffering from more attention problems and overactive behavior waited significantly shorter in the DoG task. Amniotic testosterone measures were reliable in boys only. Most importantly, boys who waited shorter in the DoG task and boys who were reported to suffer from more attention problems and overactive behavior had higher prenatal testosterone levels. These findings extend our knowledge concerning organizational effects of testosterone on the brain circuitry underlying self-control in boys, and are of relevance for understanding how sex differences in behavioral disorders are connected with a lack of self-control. [ABSTRACT FROM AUTHOR]

In this article, two fatal cases related to the use of 3-methoxyphencyclidine (3-MeO-PCP) are described. This compound is a new psychoactive substance that belongs to the phencyclidine family. In the recent period, this dissociative drug has gained interest because of its proposal as a legally available alternative to phencyclidine in some countries. The scientific literature related to 3-MeO-PCP is very poor. Using standard ultra-performance liquid chromatography-mass spectrometry and ultra-performance liquid chromatography-tandem mass spectrometry, the authors focused on the detection of 3-MeO-PCP and its metabolites in human urine. 3-MeO-PCP metabolism was studied in vitro after drug incubation with human liver microsomes and the identified metabolites were investigated in the urine of the two forensic cases. 3-MeO-PCP metabolites, including O -demethyl-3-MeO-PCP, piperidine-hydroxy-3-MeO-PCP, O -demethyl-piperidine-di-hydroxy-3-MeO-PCP and piperidine-di-hydroxy-3-MeO-PCP, were detectable in the urine from both cases and the ratio between metabolites and parent 3-MeO-PCP, always lower than 1, were calculated to estimate the proportionality of metabolites. At this stage, one can conclude that testing for 3-MeO-PCP metabolites does not increase the window of detection of the drug. [ABSTRACT FROM AUTHOR]

The authors report the case of a 41-year-old man known as a polydrug addict and found dead at home. The initial toxicological screening was inconclusive for a toxic death. However, 7 small bags of unknown powders and crystals, supposed to be NPS, were found near the body and sent to the laboratory 3 weeks later. This article focuses on the interest of nuclear magnetic resonance (NMR) spectroscopy to solve this fatal case. The 7 items were identified by 1H NMR and their purities were measured by the quantitative NMR (qNMR) method using maleic acid as internal standard. The compounds were 4-chloro-α-pyrrolidinopentiophenone, dibutylone, 4-chloroethcathinone, 4-α-pyrrolidinohexuiphenone, 3,4-methylenedioxy-N-benzylcathinone, 3-methoxy-phencyclidine and α-pyrrolidinopropiophenone, with purities in the range 51-89%. In the biological specimens of the victim, only 3-methoxy-phencyclidine (3-MeO-PCP) was identified in cardiac blood, peripheral blood and urine at 743, 489 and 16,700 ng/mL. Hair (6 cm) concentration of the drug was 15,600 pg/mg. The other NPS were not detected. The peripheral blood concentration was higher than the values reported in previous intoxicated and fatal cases. The toxicological significance of the measured concentration in hair was difficult to establish because this is only the second case describing a test result for 3-MeO-PCP. [ABSTRACT FROM AUTHOR]

The abuse of new psychoactive substances (NPS) has been dramatically increasing all around the world since the late 2000s. The availability of hundreds of NPS in the past decade is challenging for both public health and global drug policies. A 39-year-old woman, known as a multidrug addict, was murdered by her partner by ligature strangulation. A comprehensive toxicological screening by gas chromatography and ultra-high performance liquid chromatography with tandem mass spectrometry revealed the simultaneous presence of ethanol (1.37 g/L), diazepam (157 ng/mL) and nordiazepam (204 ng/mL), cocaine (25 ng/mL) and benzoylecgonine (544 ng/mL), and (3-methoxy-(1-(1-phenylcyclohexyl)piperidine) or 3-MeO-PCP, a dissociative hallucinogen anesthetic drug. Concentrations of 3-MeO-PCP were 63, 64, and 94 ng/mL in femoral blood, bile, and urine, respectively. Hair tested also positive for 3-MeO-PCP on 3 × 2-cm segments at 731, 893, and 846 pg/mg, indicating long-term abuse of the drug. This seems to be the first ever reported hair concentrations. Major impairment of the victim, including visual hallucinations and alteration of behavior, was attributed to the mixture of all the drugs, with a major contribution of 3-MeO-PCP. The toxicological findings were compared to the few reports available in the medical literature. [ABSTRACT FROM AUTHOR]

Objective: Nowadays, post-mortem computed tomography (PMCT) has become an integral part of Forensic practice. The purpose of the study was to determine PMCT impact on diagnosis of the cause of death within the context of the external examination of the body, when autopsy has, at first, not been requested.Material and Methods: We reviewed the records of 145 cases for which unenhanced PMCT was performed in addition to the external examination of the body from January 2014 to July 2015 at the Institute of Forensic medicine in Strasbourg (France). We confronted final reports from forensic pathologist to the corresponding PMCT reports. Data were collected in a contingency table and the impact of PMCT on the final conclusions of the forensic pathologist was evaluated via a Chi2 test.Results: PMCT results significantly impact the final conclusions of forensic pathologist (p<0,001). In some cases, PMCT permits etiological diagnosis by revealing a cause of death hidden from external examination (mainly natural death) or by supporting the clinical findings of the forensic pathologist. In other cases (traumatic death), PMCT enables fast and exhaustive lesion assessment. Lastly, there are situations where PMCT may be ineffective (intoxication, hanging or some natural deaths).Conclusion: Performing PMCT within the context of the external examination of the body when autopsy has, at first, not been requested could bring significant benefits in diagnosing the cause of death. The impact of PMCT varies depending on the circumstances of death. [ABSTRACT FROM AUTHOR]