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- Suicide by self-administration of pethidine, propofol and anxiolytics : a case reportPublication . Proença, Paula; Castanheira, Alice; Castanheira, Fernando; Monteiro, Carla; Simoes Da Silva, Beatriz; Franco, João Miguel; Corte Real, FranciscoBackground and aim: Recreational use of anesthetic and sedative agents in health care practitioners is becoming an increasing problem due to its easy availability and, therefore, they become vulnerable to misusing or abusing of these drugs. The authors present a fatal case involving a a 50-year-old man, nurse, found dead in a hospital room, in supine position, with intravenous access devices in the forearm that were attached to two packages of saline solutions. Some empty packages were close to the victim. A white viscous fluid found at the death scene was send to analysis. The expert's investigation at the scene indicated possible propofol poisoning. Methods: The blood samples were initially subjected to a qualitative analysis. A systematic toxicological drug screening was carried out in blood with a combination of enzyme immunoassays (ELISA) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Femoral blood alcohol concentration was measured by gas chromatography with headspace (GC/FID) technique. Due to suspected propofol intoxication, a specific analytical methodology was developed in order to detect and quantify this substance in the postmortem samples. Chromatographic analysis of propofol was preceded by protein precipitation of 0.1 mL of blood samples with acetonitrile. Chromatographic separation was achieved with a reversed-phase column, using a gradient elution of 2 mM aqueous ammonium formate (0.1% formic acid), and 2mM ammonium fluoride in methanol, at a flow rate of 0.4 mL/min, with a run time of 10 minutes. Results and discussion: Toxicological results revealed the presence of the anesthetic (propofol) in femoral blood (33.5 mg/L) and heart blood (113.3 mg/L), and also in the white viscous fluid found at the death scene. The remaining laboratory analyses revealed the presence of nordiazepam, temazepam and metoclopramide in therapeutic concentrations, but toxic blood concentrations of midazolam (1473 μg/L), diazepam (5182 μg/L) and pethidine (8622 μg/L). The blood alcohol analysis result was positive (0.18 g/L) as well as positive for opiates (morfine < 25 μg/L). Pesticides were not present in the analysed samples. Conclusion: The development of the method for confirmation and quantification of the anesthetic propofol was crucial given the suspicion of poisoning by this substance. However, laboratory analysis has also revealed the simultaneous presence of an opioid analgesic agent (pethidine) and anxiolytics in high blood concentrations. These findings have allowed to identify a case where several drugs were abused and whose synergy of effects and intoxication, that were not considered at first, were lately found to have contributed to the medico-legal etiology of suicide.
- A retrospectÍve analysis of data from cannabinoid forensic cases in the Centre of Portugal between 2020 and 2023Publication . Proença, Paula; Teixeira, Helena M.; Monteiro, Carla; Monsanto, Paula; Coelho, Maria; Franco, João Miguel; Corte Real, FranciscoBackground and aim: Marijuana, hashish and other psychoactive products obtained from Cannabis are the most produced and trafficked illicit drugs around the world. Methods: The authors studied all the positive cases for cannabinoids in the Centre of Portugal; from autopsies at the Forensic Clinical and Pathology Service of Coimbra’s Delegation of the National Institute of Legal Medicine and Forensic Sciences (NILMFS), from other autopsies in the Centre of Portugal, and road traffic and labour accidents from June 2020 until March 2023. Results were from blood samples routinely tested for the presence of cannabinoids by enzyme immunoassay (ELISA) and confirmed by LC-MS/MS. When requested, other substances were analysed. Results and discussion: During the study period, a total of 4599 toxicological requests for cannabinoids were received at the Forensic Toxicology Laboratory. 461 cases (10.0%) tested positive in screening tests, all confirmed by LC-MS/MS. Among positive cases, 77% were road traffic surveillance cases , and 24.9% related to other antemortem and postmortem cases.We observed that there was an accentuated consumption of cannabinoids between ages 21 and 30 years old, average age 33 years, and a higher prevalence in men (92.4%). THC was confirmed and quantified in 76.5% of the cases, with an average blood concentration of 13.5 ng/mL; the mean and percent for the active metabolites, 11-OH-THC, was 5.9 ng/mL in 44.5% and for THCCOOH was 20.1 ng/mL in 56.2% of the positive cases. It is important to highlight that in the 75.1% positive road traffic surveillance cases the achieved THC average was 11.6 ng/mL and in postmortem cases related to road traffic accidents this value increased to 16.0 ng/mL. 25.4% of the cases (n=117) had cannabinoids together with other psychoactive substances and ethanol was present in 162 of the cases, with 33.9% presenting a BAC ≥0.5 <1.2 g/L and 50.6% a BAC ≥1.2 g/L. 20.9% of the Forensic Pathology and Clinic cases were road accidents with 79.2% drivers. Conclusion: Cannabis is frequently used before and during driving with high concentrations of the active compounds demonstrating impairment increasing theof a crash. It is also important to be aware of the results showing simultaneous use of THC and alcohol increasing accident risk.
- Determinação de rivastigmina, inibidor seletivo da colinesterase cerebral, em amostras de sangue. Aplicação a casos forenses.Publication . Proença, Paula; Castanheira, Alice; Mustra, Carla; Castanheira, Fernando; Franco, João Miguel; Corte Real Gonçalves, FranciscoIntodução: Sempre que ocorre uma situação relevante de pesquisa de um fármaco não incluído no leque de substâncias detetadas, o laboratório dedica-se a implementar a sua técnica de modo a poder ser dada uma resposta à perícia solicitada. A rivastigmina é um inibidor das colinesterases do tipo carbamato, utilizada no tratamento de demência associada à doença de Alzheimer e à doença de Parkinson. Material e Métodos: No procedimento analítico de determinação de rivastigmina, as amostras biológicas foram previamente extraídas por precipitação proteica, utilizando 0,1 mL de amostra, e posteriormente analisadas por LC-MS/MS. A separação cromatográfica foi efetuada com uma coluna de fase reversa e uma fase móvel, em gradiente, constituída por formato de amónia aquoso 2mM e por metanol, a um fluxo de 0,4 mL/min. Na análise por MS/MS foi utilizado um triplo quadrupolo com ionização por electrospray (ESI). A aquisição de dados por monitorização de reações múltiplas (MRM) foi utilizada na deteção e quantificação de rivastigmina (RT= 4,22), de acordo com as seguintes transições: m/z 251,2>206,1 e m/z 251,2>86,2, em modo ESI positivo. Resultados e Discussão: Na validação da técnica analítica apresentada foram avaliados os parâmetros incluídos no procedimento interno de validação de métodos qualitativos e quantitativos em vigor no laboratório de toxicologia forense. Para a quantificação de rivastigmina foi utilizada uma curva de calibração com concentrações compreendidas entre 5 a 100 ng/mL e controlos ajustados a essa gama de trabalho. Foi ainda avaliada a precisão a dois níveis, a 10 e a 50 ng/mL, a precisão intermédia e o limite de repetibilidade (<20%). Quanto à exatidão, a rivastigmina apresenta recuperações médias estatisticamente semelhantes a 100%, comprovando a inexistência de erro sistemático. O método foi aplicado a 8 casos forenses com concentrações sanguíneas de rivastigmina entre 8,1 a 78 ng/mL. Os resultados obtidos da análise de rivastigmina por procedimentos de triagem, de confirmação e de quantificação foram avaliados. Conclusões: O método validado demonstrou ser seletivo, específico, exato, preciso, robusto e linear na gama de trabalho, cumprindo todos os critérios de aceitação exigidos na determinação deste tipo de substâncias em amostras biológicas analisadas no âmbito da toxicologia forense.