Search

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]

Pediatric poisoning represents a serious problem all around the world. Abuse or neglect of children by adults must be highlighted in children exposed to drugs to which they would not normally have access. Usually, segmental hair analysis would allow in these contexts to determine whether the exposure was unique or repetitive. Hair and nail samples from a 9‐month‐old girl were received in our laboratory for analysis, after the child was hospitalized due to severe dehydration caused by her mother's neglect. At the admission, flecainide, an antiarrhythmic never prescribed to the child, was identified in the daughter urine. Using an LC–MS/MS method, flecainide tested positive in the child's hair at the following concentrations: 66 pg/mg (root to 1 cm), 61 pg/mg (1–2 cm), and 125 pg/mg (2–3 cm). Traces below the limit of quantification (1 pg/mg) were also present in the nail clippings. These concentrations are much lower than those obtained in adults under daily treatment. Given the different pharmacokinetic and dynamic parameters in children, the different rate of hair growth, and the greater porosity of the hair, which makes it more prone to external contamination, the interpretation of hair findings in children remains very complicated. In this case, it can be assumed that the presence of the drug in the urine indicates systemic incorporation and that administration had occurred for some months (three positive segments). The interpretation of hair tests from young children needs a global review of all the findings, as a positive result cannot stand alone to claim repetitive exposures. [ABSTRACT FROM AUTHOR]

Sodium‐glucose cotransporter 1 inhibitors are a new class of drugs used for the treatment of type II diabetes. Due to their diuretic capabilities and the glycosuria they induce, these molecules cause effective weight loss that could attract the interest of a wider public than diabetics with all the health consequences knowing the adverse effects of these substances. In order to reveal a past exposure to these substances, hair analysis can be very useful especially in the medicolegal context. There are no data in the literature about gliflozin testing in hair. In this study, a method was developed for the analysis of three molecules belonging to the gliflozin family (dapagliflozin, empagliflozin and canagliflozin) using a liquid chromatography system coupled to tandem mass spectrometry. After decontamination with dichloromethane, gliflozins were extracted from hair following incubation in methanol in the presence of dapagliflozin‐d5. Validation showed acceptable linearity for all compounds between 10 and 10,000 pg/mg, with limit of detection and limit of quantification at 5 and 10 pg/mg, respectively. Repeatability and reproducibility were below 20% at three concentrations for all analytes. The method was subsequently applied to the hair of two diabetic subjects under dapagliflozin treatment. In one of the two cases, the result was negative, while in the second case, the concentration was 12 pg/mg. Due to the absence of data, it is difficult to explain the absence of dapagliflozin in the hair of the first case. Physico‐chemical characteristics of dapagliflozin could explain its bad incorporation in hair, making detection difficult even after daily treatment. [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]

Early November 2020 an Olympic gold medal winner returned during an out‐competition control an adverse analytical finding for dorzolamide, a diuretic mostly used to treat glaucoma. Estimated urine concentrations were 2.2 and 1.6 ng/ml in the A and B specimens, respectively. As the athlete denied any use of dorzolamide, a complex forensic investigation was suggested. It revealed that the athlete was severely injured during a car crash 6 months before where he received 2× 500 ml of red blood cells transfusion. One of the blood donors declared using dorzolamide. A plasma aliquot, stored for legal purposes, was tested by LC‐MS/MS several months later and contained 4.3 ng/ml of dorzolamide. Given the very long half‐life of the drug, up to 150 days and its reported incorporation into erythrocyte, it was accepted by the French antidoping administration (AFLD) that the source of contamination was this blood transfusion and that the antidoping rule violation was unintended. [ABSTRACT FROM AUTHOR]

Tizanidine, an imidazoline derivative close to clonidine, is a central alpha‐2 adrenergic receptor agonist. Therapeutically, the drug is used as a muscle relaxant under the trade names Sirdalud™ or Zanaflex™. The drug is not prohibited by the World Anti‐Doping Agency but, for therapeutic purposes, can only be obtained via a nominative temporary use authorization. The French public health police requested the laboratory to test for tizanidine in head hair specimens collected from international racing cyclists. Using Liquid chromatography–tandem mass spectrometry (LC/MS–MS) and confirmation by liquid chromatography–high‐resolution mass spectrometry (LC/HRMS), after pH 9.5 borate buffer overnight incubation of 20 mg and subsequent solvents extraction, tizanidine was identified in the hair of three athletes at 1.1, 3.7, and 11.1 pg/mg. This is the first evidence that tizanidine is incorporated in human hair. However, it was not possible to interpret the data in terms of doses and frequency of use due to a lack of controlled study. [ABSTRACT FROM AUTHOR]

Since the late 90s, hair testing for anabolic steroids in humans has found numerous forensic, clinical, and anti‐doping applications. In most cases, analyses were performed on head hair, collected in the vertex regions. However, for various reasons (shaved subject, bald subject, religious belief, cosmetic treatment and aesthetic reason), hair collectors can face the lack of head hair, and therefore, body hair can be the unique alternative choice. Although there is no possibility to perform segmental analyses with body hair, their use has two major advantages: (1) In most cases, anabolic steroids are more concentrated in body hair when compared with head hair, which allows detecting abuse at lower frequency and for lower dosages; and (2) the window of drug detection is generally much longer in body hair when compared with head hair, particularly in male athlete presenting short head hair. To document the relevance of simultaneous collection of head and body hair, the authors present eight authentic cases of anabolic steroids abuse, including clostebol (one case), drostanolone (one case), metandienone (one case), 19‐norandrostenedione (one case), stanozolol (two cases) and trenbolone (three cases). In all cases, body hair concentrations were higher than head hair concentrations. Even in three cases, no steroid was identified in head hair, although present in body hair. [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]

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]

For 40 years, hair tests have been presented as the best approach to document long‐term consumption of a drug. This unique property has found numerous applications in clinical, forensic, and occupational toxicology. However, since the beginning of its implementation in biology, external contamination, with an associated risk of false positive result, has been presented as the key in the final interpretation. Evidence of environmental contamination and subsequent health issues can be the task of any toxicologist. Because of recent progress of analytical equipment, it is now possible to quantify drugs in hair with high level of accuracy and specificity at the pg/mg range. Therefore, segmental hair tests can be used to document environmental contamination and are the objective of this publication. In a family law case, N,N‐dimethyltryptamine (DMT), a powerful hallucinogen, has been found in the hair of the partner of a repetitive DMT smoker at 4 to 13 pg/mg in 6 × 1 cm segments, with a regular increase of concentrations from the proximal to the distal hair end. The low measured concentrations and the particular pattern of DMT distribution along the hair shaft seem to be typical of environmental contamination, the older hair (those of the distal part) being for a longer time in contact with the drug. Despite strong decontamination, drugs from the environment can remain bound to the hair matrix and therefore be able to be used to document environmental contamination. [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]

Ostarine, also known as S22 or MK2866 and enobosarm, is a selective androgen receptor modulator (SARM). It has high anabolic potency, in addition to limited androgenic effects. At this time, ostarine has no therapeutic use, but can be abused for performance‐enhancing purposes using the oral route, at dosages of 10–25 mg per day. As the drug can easily be obtained via the Internet or some fitness centers, athletes and more and more amateurs can use it without undergoing the deleterious physiological side effects that are generally associated with testosterone‐related compounds. Since 2008, the World Anti‐Doping Agency (WADA) has prohibited SARMs at‐all‐times in the category of "other anabolic agents" under section S1.2 of the WADA List. In a case of trafficking/abuse, ostarine was identified in nail (subject 1) and hair (subject 2) by LC/HRMS after incubation of 50 mg of matrix in a pH 9.5 buffer, followed by extraction with organic solvents. The drug was quantitated by LC–MS/MS. Ostarine tested positive at 61 pg/mg (toenails) and 111 pg/mg (fingernails) for subject 1. Ostarine was present at 146, 168, 93, and 101 pg/mg in the 4 × 3 cm hair sections of subject 2, clearly demonstrating long‐term use. [ABSTRACT FROM AUTHOR]

GW501516, also known as GW‐1516 or cardarine and endurobol, is a peroxisome proliferator‐activated receptor delta (PPAR‐δ) agonist. Activation of the receptor will increase fat‐burning capacity and muscle production, as it changes the body's fuel preference from glucose to lipids. GW501516 has no therapeutic use, but can be abused for performance‐enhancing purposes using the oral route, at dosages of 10 to 20 mg per day, for 6 to 8 weeks. Both athletes and amateurs can abuse GW501516 as the drug can be easily obtained via the Internet. Since January 2009, the list of prohibited substances and methods of doping as established by the World Anti‐Doping Agency includes GW‐501516, first as a gene doping substance and now in the S4.5 Metabolic modulators class. It is prohibited at all times. Using LC/MS–MS and confirmation by LC/HRM, after methanol incubation of 20 mg with ultrasound for 1 hour, GW501516 was identified in the hair of a male abuser at 32 and 22 pg/mg in 2 × 2 cm segments. The result is the first evidence that this compound with a carboxylic acid function is incorporated in human hair. [ABSTRACT FROM AUTHOR]

The stability of compounds in formalin solution is an important factor for drug analysis in a toxicological investigation. In this article, the authors report a complex medico‐legal case involving midazolam and oxycodone. The complexity of this case comes from the fact that the body was embalmed with formalin solution before the autopsy. This technique, called thanatopraxy, allows the preservation of corpses from decomposition, the destruction of a maximal number of micro‐organisms, and the presentation of the body with a natural appearance to the family. Unfortunately, when thanatopraxy is performed before the collection of biological specimens, the toxicological results are not representative of the time of the death. In addition, the interpretation of the results is difficult, because formalin can cause oxidation of xenobiotics present in the body at the time of the death, alter the pH of the tissues and dilute the compounds. To document the chemical stability of midazolam and oxycodone in formalin solution and interpret the results, a stability study was conducted for 21 days. Blood containing midazolam and oxycodone was spiked with formalin, kept at 4°C and regularly tested for both drugs. This study showed a rapid degradation of midazolam and oxycodone (85% during the first 24 hours for oxycodone). In the peripheral blood of the victim, methanol (1.31 g/L), midazolam (74ng/mL) and oxycodone (152 ng/mL) were identified. According to the stability study, the measured concentrations in formalin fixed‐tissues are to be interpreted very carefully, knowing that significant degradation has occurred. [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.

Designer benzodiazepines have emerged as recreational drugs. They are available via the Internet without control and are found in the form of falsified (fake) medicines. For some of them, limited information concerning their effects, their toxicity, and their detection in bio fluids is available in the literature. For others, nothing has been published, as in the case of flunitrazolam (FNTZ). To gain preliminary data on its elimination parameters in urine and to investigate its metabolism, one of the authors ingested one pink tablet bought on the Internet, after confirming the absence of other compounds and agreement with the labeled dosage (0.25 mg) by nuclear magnetic resonance (NMR). A software algorithm (MetaboLynx, Waters, Milford, MA, USA) was used to predict FNTZ biotransformation and four potential metabolites were proposed: 4‐hydroxy‐FNTZ, desnitro‐FNTZ, 7‐amino‐FNTZ, and 7‐acetamido‐FNTZ. Urine samples were collected over 72 hours following oral administration of one tablet. After liquid/liquid extraction at pH 9.5, FNTZ concentrations were determined using ultra performance liquid chromatography–triple quadrupole–mass spectrometry (UPLC–QqQ–MS/MS). FNTZ remained detectable in hydrolyzed urine for 21 hours after ingestion, with concentrations ranging between 1 and 18 ng/mL. About 3% of the initial dose was excreted in urine as total unchanged FNTZ during this period. In vitro experiments (HLM incubations) were performed using ultra performance liquid chromatography–quadrupole time of flight‐mass spectrometry (UPLC–QTOF–MS) in order to investigate the potential CYP‐ and UGT‐dependent metabolites where only 7‐amino‐FNTZ was detected as the only metabolite. However, in the urine specimens, desnitro‐FNTZ, 7‐acetamido‐FNTZ and 7‐amino‐FNTZ were the main detected compounds. The identification of FNTZ metabolites dramatically improves the detection windows of the drug up to 37 hours. Flunitrazolam is a designer benzodiazepine for which no information is available in the scientific literature. In order to gain data on its elimination in urine and to investigate its metabolism, one subject ingested one tablet (0.25 mg) bought on the Internet. Flunitrazolam remained detectable in hydrolyzed urine for 21 hours and its metabolites (7‐amino‐ flunitrazolam and 7‐acetamido‐flunitrazolam) improve the detection windows of the drug up to 37 hours. [ABSTRACT FROM AUTHOR]

Designer benzodiazepines provide an attractive alternative to prescribed benzodiazepines for abuse purposes as they are readily available via the Internet without control. Metizolam was ordered via the Internet and a 2 mg blue tablet was orally administered to a 54-year-old man. Urine samples were collected over 6 days in polypropylene tubes. After liquid/liquid extraction at pH 9.5, metizolam was analyzed by ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS) using a standard method devoted to benzodiazepines, and ions transitions, at m/ z 328. 9 > 275. 0 and 328. 9 > 300. 0. Metizolam was detectable in hydrolyzed urine during the 46-h period, with concentrations always lower than 11 ng/mL. About 0.3% of the initial dose was excreted in urines as total unchanged metizolam during the first 24 h. The most relevant potential CYP- and UGT-dependent metabolites of metizolam were investigated in vitro using human liver microsome incubation and, subsequently, liquid chromatography coupled with quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF-MS) analysis. Three mono-hydroxylated metabolites were produced including a hydroxylation compound at the 2-ethyl moiety of metizolam (M1) as quantitatively main metabolite, and a N-hydroxymetiazolam (M2). The structure of the third metabolite (M3) could not be elucidated because of a too low experimental production rate. Two authentic urine samples were analyzed using the same analytical method to search for metabolites of metizolam. M1, together with its glucuronide (M1-Glu), and M2 were observed in urine at the 8 h mark, whereas only M1 and M1-Glu were still detected in urine at 30 h post administration. Copyright © 2016 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

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]

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.