Two independent monoclonal antibodies were generated against psilocybin and its dephosphorylated metabolite psilocin, the psychoactive compounds in hallucinogenic mushrooms. Novel immunogenic conjugates were prepared by modifying the side chains of these molecules and linking them to carrier proteins. Mice were immunized, and hybridoma clones secreting the specific antibodies were established. Competitive enzyme-linked immunosorbent assays (ELISAs) using these antibodies enabled detection of psilocybin and psilocin at ranges of approximately 0.20–20 μg/assay and 0.040–2.0 μg/assay, respectively, with low cross-reactivity between the two compounds. In dried Psilocybe cubensis powder, psilocybin and psilocin contents were 0.39% and 0.32% by weight. These ELISAs offer a promising tool for identifying illegal hallucinogenic mushrooms.
A new test detects psilocin, the main psychoactive compound in hallucinogenic mushrooms, with much higher sensitivity than before. The method first converts psilocin into a heavier chemical form (TBS/Psi), then uses an antibody that binds strongly to this modified compound. The antibody showed 69-fold higher affinity than an earlier version, and the test's detection midpoint was over 100-fold lower than the previous assay, reaching the desired low-picomole sensitivity. When applied to dried Psilocybe cubensis mushroom powder, the test gave positive signals indicating expected psilocin levels, while four edible mushroom species produced no detectable response.
Japan's Ministry of Health, Labor and Welfare amended the Pharmaceutical Affairs Law in 2006 to control 31 non-controlled psychotropic substances and one plant as "Designated Substances" as of April 2007, with five more compounds added in January 2008. These substances had been sold as video cleaners, incense, and reagents via the Internet or in video shops. The authors developed simultaneous analytical methods using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) and present retention times, UV spectra, and mass spectrometry data for these substances.