Phase I metabolites (organic acids) of gamma‐hydroxybutyric acid–validated quantification using GC–MS and description of endogenous concentration ranges

Gamma-hydroxybutyric acid (GHB) is a sedative drug used in drug-facilitated crimes. Its detection window is very short. GHB undergoes intensive phase I metabolism to organic acids (glycolic acid, succinic acid, dihydroxybutyric acids). These could be potential analytical targets to broaden the detection window. The aim of the present study was to enable the detection of endogenous levels of these metabolites in biological samples (blood and urine). A gas chromatographic–mass spectrometric method using liquid–liquid extraction and derivatization with N-methyl-N-tri-methylsilyltrifluoracetamide was developed for the quantification. Validation results were consistent with international guidelines, and the method was able to quantify endogenous levels of the substances in both urine and blood. Endogenous concentrations were shown to be <0.03–4.92 mg/L for glycolic acid, <0.03–1.28 mg/L for GHB, <0.28–18.1 mg/L for succinic acid, <0.12–1.38 mg/L for 2,4-dihydroxybutyric acid, and <0.13–2.59 mg/L for 3,4-dihydroxybutyric acid in serum samples of 101 volunteers. Urinary endogenous concentrations were shown to be 1.30–400 mg/L for glycolic acid, <0.03–1.94 mg/L for GHB, 1.17–2.73 mg/L for succinic acid, 0.72–26.2 mg/L for 2,4-dihydroxybutyric acid, and 1.88–122 mg/L for 3,4-dihydroxybutyric acid in urine samples of 132 volunteers. These endogenous concentrations represent a basis to which concentrations after the intake of GHB can be compared to in order to prove the intake of this substance

Identification of S22 (ostarine) in human nails and hair using LC‐HRMS. Application to two authentic cases

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

Fatal inhalation of nitrogen inside a closed environment: toxicological issues about the cause of death

Asphyxia due to inhalation of nitrogen, as accidental or suicidal event, has been seldom reported in the forensic field. Death usually occurs because of a displacement and decrease of environmental and alveolar oxygen concentrations, but taking into account that nitrogen is a normal component of atmosphere, autopsy findings, which may be in certain cases unremarkable, must be corroborated with a careful scene investigation in order to determine the cause of death. We present a case of a 37-year old male found unresponsive inside his car with five liquid nitrogen tanks placed in the trunk, three of them with clear losses because of incomplete lock. Resuscitation efforts were unsuccessful. Autopsy findings and histological analyses were unremarkable, but toxicological analyses were crucial. Percentages of nitrogen were high in blood collected from the inferior vena cava (85.2%), left ventricle (81.01%) and the right lung (80.73%). Concentrations of nitrogen were higher than those detected in control samples: 14 autopsy cases, room air and water. The cause of death was identified as an inert gas asphyxiation, which was classified as accidental in accordance with the police report