RESUMEN
Ethanol is the most frequently psychoactive substance used in the world, leading to major public health problems with several millions of deaths attributed to alcohol consumption each year. Metabolism of ethanol occurs mainly in the liver via the predominant oxidative metabolism pathway involving phase I enzymes including alcohol dehydrogenases (ADH), cytochrome P450 (CYP) 2E1 and catalase. In a lesser extent, an alternative non-oxidative pathway also contributes to the metabolism of ethanol, which involves the uridine diphospho-glucuronosyltransferase (UGT) and sulfotransferase (SULT) phase II enzymes. Using liquid chromatography-high resolution mass spectrometry, ethylglucuronide (EtG) and ethylsulfate (EtS) produced respectively by UGT and SULT conjugation and detected in various biological samples are direct markers of alcohol consumption. We report herein the efficient non-oxidative metabolic pathway of ethanol in human differentiated HepaRG cells compared to primary human hepatocytes (HH). We showed dose- and time-dependent production of EtS and EtG after ethanol (25 or 50â¯mM) treatment in culture media of differentiated HepaRG cells and HH and a significant induction of CYP2E1 mRNA expression upon acute ethanol exposure in HepaRG cells. These differentiated hepatoma cells thus represent a suitable in vitro human liver cell model to explore ethanol metabolism and more particularly EtG and EtS production. In addition, using recombinant HepG2 cells expressing different UGT1A genes, we found that UGT1A9 was the major UGT involved in ethanol glucuronidation.
Asunto(s)
Etanol/farmacología , Glucuronatos/metabolismo , Glucuronosiltransferasa/metabolismo , Hepatocitos/metabolismo , Células Cultivadas , Glucurónidos/metabolismo , Humanos , Sulfotransferasas/metabolismo , Ésteres del Ácido Sulfúrico/metabolismo , UDP Glucuronosiltransferasa 1A9RESUMEN
BACKGROUND: Identification of metabolites is of importance in the challenge of new psychoactive substances (NPS) as it could improve the detection window in biological matrices in clinical and forensic cases of intoxication. Considering the numerous and diverse NPS reported each year, producers increasingly appear today to be targeting non-controlled synthetic opioids, involving fentanyl derivatives such as furanyl fentanyl (Fu-F). OBJECTIVE: This work aims to investigate and compare metabolites of Fu-F using two in vitro experimental approaches. METHODS: CYP- and UGT-dependent metabolites of Fu-F were investigated by means of analyses of both human liver microsome (HLM) and hepatic (HepaRG) cell line incubates using liquid chromatography with high-resolution mass detection and, subsequently, compared and confronted to recently published data. RESULTS: Seventeen Fu-F metabolites were produced and several metabolic pathways can be postulated. HLMs and HepaRG cultures appear to be complementary: HepaRG cells produced 9 additional metabolites, but which appear to be minor in vivo metabolites. Specific* and/or abundant Fu-F metabolites are dihydrodiol-Fu-F*, norFu-F* and despropionylfentanyl. However, norFu-F seems to be inconstantly observed in in vivo cases. Furthermore, a sulfate metabolite presents at significant rate in urine obtained from FU-F users was not identified here, as in another in vitro study. CONCLUSION: HLMs represent an acceptable first choice tool for a single NPS metabolism study in forensic laboratories. Dihydrodiol-Fu-F and despropionylfentanyl could be proposed as reliable metabolites to be recorded in HRMS libraries in order to improve detection of Fu-F users. Nevertheless, additional verifications of in vivo data remain necessary to confirm relevant blood and urinary metabolites of Fu-F.
Asunto(s)
Analgésicos Opioides/análisis , Fentanilo/análogos & derivados , Furanos/análisis , Microsomas Hepáticos/metabolismo , Detección de Abuso de Sustancias/métodos , Analgésicos Opioides/metabolismo , Biomarcadores/metabolismo , Biotransformación , Técnicas de Cultivo de Célula , Línea Celular Tumoral , Cromatografía Liquida , Fentanilo/análisis , Fentanilo/metabolismo , Furanos/metabolismo , Humanos , Espectrometría de MasasRESUMEN
Powdered roots of iboga (Tabernanthe iboga) contain ibogaine, an alkaloid that has been used to treat addictions. We report the case of a 30-year-old woman who died after ingesting a powder labeled as Tabernanthe iboga she had bought online. Analysis of the powder revealed the absence of ibogaine but the presence of toxic alkaloids (ajmaline, yohimbine and reserpine) found in Rauvolfia sp. plant species. An original and specific LC-MS/MS method developed to quantify ajmaline, yohimbine and reserpine showed respective concentrations of 109.1ng/mL, 98.2ng/mL and 30.8ng/mL in blood, and 1528.2ng/mL, 914.2ng/mL and 561.2ng/mL in bile. Moreover, systematic toxicological analyses of biological samples showed the presence of oxazepam at therapeutic concentration and cannabinoids. Death could be attributed to ingestion of a substantial quantity of crushed roots of Rauvolfia in association with concomitant drug withdrawal.
Asunto(s)
Etiquetado de Productos , Rauwolfia/envenenamiento , Alcaloides de Triptamina Secologanina/envenenamiento , Adulto , Cromatografía Liquida , Femenino , Toxicología Forense , Humanos , Polvos , Alcaloides de Triptamina Secologanina/aislamiento & purificación , Espectrometría de Masas en TándemRESUMEN
Chlormequat is a quaternary ammonium used as plant growth regulating agent. We report here the first suicide case involving a 45 year-old farmer man who intentionally self-injected C5SUN(®), containing chlormequat and choline. An original liquid chromatography high resolution mass spectrometry method (LC-HR-MS), using a hybrid quadrupole-orbitrap mass spectrometer, was developed for qualitative and quantitative analysis of chlormequat in different biological matrices. Toxicological analyses of post-mortem samples highlighted the presence of chlormequat in the blood (2.25mg/L) and the urine (4.45mg/L), in addition to ethanol impregnation blood (1.15g/L). The route of administration (subcutaneous injection) was confirmed by the detection of chlormequat in the abdominal fat sample (chlormequat: 10.04mg/g) taken from the traumatic injury location, as well as in the syringe found at the death scene, close to the victim's body. Based on the results of these post-mortem investigations, the cause of death was determined to be consecutive to cardiac dysrhythmia and cardiac arrest following chlormequat self-injection.