Journal of Chromatography B: Biomedical Sciences and Applications
Determination of psilocybin in Psilocybe semilanceata by capillary zone electrophoresis
Introduction
Psilocybe semilanceata is the most abused hallucinogenic mushroom in the Scandinavian countries. Dried mushrooms have been shown to contain up to 2% of psilocybin (Fig. 1A) which is the major active constituent. The dephosphorylated compound, psilocin (Fig. 1B), is only found in trace amounts. However, up to 0.3% of the demethylated derivative of psilocybin, baeocystin (Fig. 1C), has been detected [1]. In a study of mushrooms from several European countries, Psilocybe semilanceata and Panaeolus subbalteatus proved to be the only psilocybin-containing fungi that can be gathered in Middle and Northern Europe in sufficient quantities to permit abuse [2]. However, psilocybin has been detected worldwide in a variety of mushrooms belonging to the genera of Psilocybe, Panaeolina, Panaeolus, Copelandia, Conocybe, Gymnopilus, Stropharia and Pluteus 3, 4. Some recent reports include the detection of psilocybin in mushrooms from Thailand [5], the Venezuelan Andes [6], the Hawaiian islands [7]and Thailand [8].
Since psilocybin is a controlled compound and considered a narcotic drug, it must be reliably identified and quantified in mushroom samples. Identification of psilocybin, psilocin and baeocystin has been carried out by thin-layer chromatography 2, 9. Due to their high polarity and amphoteric nature, high-performance liquid chromatography (HPLC) either in the normal-phase mode [10]or in the reversed-phase mode [11]has been a preferred method for their quantification, utilizing either UV, fluorescence or electrochemical detection [12]. Gas chromatography has been carried out after derivatization [9], and GC–MS analysis has been performed for recording of mass spectra for court evidence [13].
Capillary electrophoresis (CE) has been used increasingly to analyse seized drugs, and a variety of methods has been worked out for narcotic drug analysis [14]. CE can analyse drugs which are nonvolatile, polar and thermally degradable, and would in principle be an excellent technique for the analysis of psilocybin and baeocystin in mushroom samples. Psilocybin has previously been separated from other narcotic drugs in a MEKC forensic drug screen using SDS as surfactant [15]. No CE method has, however, been reported for the separation of the closely related compounds psilocybin and baeocystin in hallucinogenic mushrooms. Therefore, the aim of the present study was to develop a CE method for the rapid separation, identification and quantification of psilocybin in mushroom samples. In order to maintain analytical simplicity, the work was carried out with capillary zone electrophoresis. Because of high importance within the European countries, the assay was focused on the mushroom Psilocybe semilanceata.
Section snippets
Chemicals
Psilocybin was supplied by Sandoz (Basel, Switzerland). Methanol of HPLC grade and analytical grades of sodium tetraborate and sodium dihydrogenphosphate were from Merck (Darmstadt, Germany). The internal standard barbital was purchased from Norsk Medisinaldepot (Oslo, Norway). Deionized water was obtained from a Milli-Q system (Millipore, MA, USA), and samples of Psilocybe semilanceata were supplied by The Bureau of Crime Investigation (Oslo, Norway).
CE system
The CE system was a P/ACE 5000 Series from
Separation considerations
As discussed in Section 1, the zwitterionic compound psilocybin (Fig. 1A) is the major constituent contributing to the hallucinogenic effect of the mushroom Psilocybe semilanceata. Therefore, the method developed in the present work was primarily focused on the rapid and unequivocal determination of this particular compound. However, Psilocybe semilanceata also contains considerable amounts of baeocystin (Fig. 1C), which is a demethylated derivative of psilocybin. Although required for a
Conclusions
The present work has demonstrated a new, fast and highly reliable method based on capillary zone electrophoresis for the determination of psilocybin in seized mushrooms. Excellent separation of psilocybin and baeocystin was obtained in free solution with a 10 mM borate–phosphate running buffer adjusted to pH 11.5. Reliable identification was based on migration times and UV spectra, while repeatable and reproducible quantification was performed utilizing barbital as internal standard. Compared
Acknowledgements
This work has been financially supported by The Department of Justice (Oslo, Norway). The Bureau of Crime Investigation (Oslo, Norway) and Professor Klaus Høiland are acknowledged for supplying the samples of Psilocybe semilanceata.
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2015, Studies in Natural Products ChemistryCitation Excerpt :These include liquid–liquid extraction (LLE), Soxhlet extraction, solid-phase extraction (SPE), or supercritical fluid extraction (SFE). The most popular extraction methods for isolation hallucinogenic substances from mushrooms are ultrasonic-assisted extraction and SPE (Table 5.1) [10,26–37]. Ultrasounds are the source of additional energy, which allows separating the analytes from the sample matrix by destroying mushroom cells.
Chapter 4 Hallucinogens
2008, Handbook of Analytical SeparationsCitation Excerpt :Another method using methanol to extract psilocybin from the plant material has been reported. This method also used CZE at pH 11.5 with barbital as internal standard giving a linear range 0.01–1.0 mg/mL [48]. Methanol was also used in a single extraction step by other investigators who concluded it to be the best for analysis of psilocybin from mushrooms [49].