A validated method for quantitation of psilocin in plasma by LC–MS/MS and study of stability

"Magic mushrooms" is the most common name given to hallucinogenic fungi containing the psychoactive alkaloids psilocybin and psilocin. In recent years, fungis' sclerotia, commonly called "magic truffles" have become a form of supply of psychoactive Psilocybe alkaloids since Psilocybe sclerotia are not specifically included in the laws banning the sale, the purchase and the use of such substances and mushrooms containing them. A liquid chromatography -tandem mass spectrometry (LC-MS/MS) method was developed for the rapid determination of psilocybin and psilocin in Psilocybe sclerotia. Following a simple step extraction with methanol, the alkaloids were separated on a reversed-phase column using a gradient of 0.1% formic acid - acetonitrile s a mobile phase at a flow rate of 0.2 mL/min.. Separated analytes were detected by electrospray ionization tandem mass spectrometry in the positive ion mode using multiple reaction monitoring. The developed method was linear over the calibration range for all two substances under investigation, with a r(2) > 0.99. The detection and quantification limits were 0.3 µg and 1 µg per 100 mg truffles, for both psilocin and psilocybin and the intra- and inter-day coefficients of variation were always better than 15%. Using this method, the presence of only psilocybin was demonstrated in examined Psilocybe sclerotia. The content of psilocybin was found to vary over a concentration range of 59.3 to 167.8 µg per 100 mg of fresh sclerotia.

Section: Resultsmentioning

confidence: 99%

Magic truffles or Philosopher's stones: a legal way to sell psilocybin?

Pellegrini

Rotolo

Marchei

et al. 2012

“…19 29 Martin et al 30 could overcome the psilocin instability in plasma by freezing. Stability of classic drugs of abuse, therapeutic drugs, and their metabolites in blood, plasma, or serum has been discussed elsewhere.…”

Section: Analyte Stabilitymentioning

confidence: 99%

“…cathinones, 26,27 piperazines, 27,28 and synthetic cannabinoids. 29 Martin et al 30 could overcome the psilocin instability in plasma by freezing. In contrast, psilocin glucuronide, the main psilocin metabolite, was stable in serum and urine and should thus be included in the approach for monitoring psilocin intake.…”

Section: Analyte Stabilitymentioning

confidence: 99%

Pitfalls in drug testing by hyphenated low‐ and high‐resolution mass spectrometry

Maurer

2020

This paper reviews various pitfalls observed during developing, validation, application, and interpretation of drug testing approaches using GC-MS and low-and high-resolution LC-MS. They include sampling and storage of body samples, sample adulteration and contamination, analyte stability, sample preparation without or with cleavage of conjugates, extraction, derivatization, internal standardization, false negative and positive results by GC-MS or LC-MS screening and/or confirmation procedures including artifact formation, ion suppression or enhancement by electrospray ionization, and finally pitfalls in data interpretation. Conclusions and prospects close the Tutorial. K E Y W O R D S

“…18 Psilocin and bufotenine glucuronide should be included as psilocin-O-glucuronide showed a higher stability in biosamples amongst others and as it can be distinguished from bufotenin-O-glucuronide via their MS 2 patterns. 18,29,30 A strategy should be developed and validated offering an easier of workflow and less turnaround time. Applicability should be tested by analyzing human urine samples obtained from suspected mushroom intoxication cases.…”

Section: Introductionmentioning

confidence: 99%

Further development of a liquid chromatography–high‐resolution mass spectrometry/mass spectrometry‐based strategy for analyzing eight biomarkers in human urine indicating toxic mushroom or Ricinus communis ingestions

Bambauer

Wagmann

Weber

et al. 2021

Recently, we presented a strategy for analysis of eight biomarkers in human urine to verify toxic mushroom or Ricinus communis ingestions. However, screening for the full panel is not always necessary. Thus, we aimed to develop a strategy to reduce analysis time and by focusing on two sets of analytes. One set (A) for biomarkers of late-onset syndromes, such as phalloides syndrome or the syndrome after castor bean intake. Another set (B) for biomarkers of early-onset syndromes, such as pantherine-muscaria syndrome and muscarine syndrome. Both analyses should be based on hydrophilic-interaction liquid chromatography coupled with high-resolution mass spectrometry (MS)/MS (HILIC-HRMS/MS). For A, urine samples were prepared by liquid-liquid extraction using dichloromethane and subsequent solid-phase extraction of the aqueous supernatant. For B urine was precipitated using acetonitrile. Method A was validated for ricinine and αand β-amanitin and method B for muscarine, muscimol, and ibotenic acid according to the specifications for qualitative analytical methods. In addition, robustness of recovery and normalized matrix factors to matrix variability measured by urinary creatinine was tested. Moreover, applicability was tested using 10 urine samples from patients after suspected mushroom intoxication. The analytes αand β-amanitin, muscarine, muscimol, and ibotenic acid could be successfully identified. Finally, psilocin-O-glucuronide could be identified in two samples and unambiguously distinguished from bufotenine-O-glucuronide via their MS 2 patterns. In summary, the current workflow offers several advantages towards the previous method, particularly being more labor-, time-, and costefficient, more robust, and more sensitive.