Development and validation of a sensitive LC-MS-MS method to quantify psilocin in authentic oral fluid samples.

Psilocin is an active substance and a dephosphorylated product of psilocybin formed after the ingestion of mushrooms. The low stability caused by the quick oxidation of this analyte requires sensitive methods for its determination in biological matrices. In this work, we described the development, optimization and validation of a method for the quantification of psilocin in authentic oral fluid samples by liquid chromatography-tandem mass spectrometry. Liquid-liquid extraction was performed using 100 µL of oral fluid samples collected with a Quantisal™ device and t-butyl methyl ether as the extraction solvent. The method showed acceptable performance, with limits of detection and quantification of 0.05 ng/mL, and the calibration model was achieved between 0.05 and 10 ng/mL. Bias and imprecision results were below -14.2% and 10.7%, respectively. Ionization suppression/enhancement was lower than -30.5%, and recovery was >54.5%. Dilution integrity bias was <14.4%. No endogenous and exogenous interferences were observed upon analyzing oral fluid from 10 different sources and 56 pharmaceuticals and drugs of abuse, respectively. No carryover was observed at 10 ng/mL. Psilocin was stable in oral fluid at -20°C, 4°C and 24°C up to 24, 72 and 24 h, respectively, with variations <17.7%. The analyte was not stable after three freeze/thaw cycles, with variations between -73% and -60%. This suggests the instability of psilocin in oral fluid samples, which requires timely analysis, as soon as possible after the collection. The analyte remained stable in processed samples in an autosampler (at 10°C) for up to 18 h. The method was successfully applied for the quantification of five authentic samples collected from volunteers attending parties and electronic music festivals. Psilocin concentrations ranged from 0.08 to 36.4 ng/mL. This is the first work to report psilocin concentrations in authentic oral fluid samples.

Is There Enough Knowledge to Standardize a Cannabis sativa L. Medicinal Oil Preparation with a High Content of Cannabinoids?

Introduction: Cannabis sativa L. medicinal oils are good therapeutic options due to their wide spectrum of pharmacological applications and the easy adjustment of individual doses. The lack of standardization of methodology in the preparation of medicinal oil using the Cannabis crude extract results in elevated variability of cannabinoid concentration in the final product. The elevated variability impairs the understanding of beneficial and adverse effects related to dose-response pharmacological activities. Objective: This study aimed to conduct a review on the current methods of Cannabis oil preparation present in the literature, to demonstrate the most appropriate methodologies to ensure a product with high content of cannabinoids and terpenes. Results: The decarboxylation stage is essential for the conversion of acid cannabinoids into neutral cannabinoids, which are substances with the highest bioavailability. Lower temperatures for longer periods of time instead of high temperatures in less time are highly recommended to ensure that all the acidic cannabinoids have passed through decarboxylation. For the guarantee of a high terpene content, the separate addition of essential oil to the fixed oil prepared from the crude extract should be considered. Ultrasound-assisted extraction is one of the best performing methodologies because it is cheaper than other techniques, such as supercritical fluid extraction, besides that, ultrasound extraction is effective in short extraction times and uses small amounts of solvent when compared with other techniques. Conclusion: Although the literature about the methods of preparation of Cannabis medicinal oil is scarce, it is possible to standardize an optimized, low-cost, and effective Cannabis extractive methodology from the results found in the literature; however, this will depend on new research for methodological validation.

Prevalence of new psychoactive substances (NPS) in Brazil based on oral fluid analysis of samples collected at electronic music festivals and parties.

BACKGROUND: New psychoactive substances (NPS) use is a worldwide public health issue. Knowing the prevalence of NPS guides public health and legal policies to address the problem. The objective of this study was to identify NPS in Brazil through the analysis of oral fluid (OF) samples collected at parties and electronic music festivals. METHODS: Anonymous questionnaires and oral fluid samples were collected from volunteers (≥18 years) who reported the consumption of at least one illicit psychoactive substance in the last 24 h. Oral fluid sample collections occurred at eleven parties and two electronic music festivals over 16 months (2018-2020). Questionnaire answers were matched to oral fluid toxicological results. RESULTS: Of 462 oral fluid samples, 39.2 % were positive for at least one NPS by liquid chromatography‒tandem mass spectrometry (LC-MS/MS). The most prevalent NPS was ketamine (29.4 %), followed by methylone (6.1 %) and N-ethylpentylone (4.1 %); however, MDMA was the most commonly identified (88.5 %) illicit psychoactive substance. More than one drug was identified in 79.9 % of samples, with two (34.2 %) and three (23.4 %) substances most commonly observed. Only 5 % of volunteers reported recent NPS consumption. CONCLUSION: MDMA is still the most common party and electronic music festival drug, although NPS were identified in more than one-third of oral fluid samples.