A method using high-performance liquid chromatography with chemiluminescence detection was developed to measure psilocin and psilocybin in hallucinogenic mushrooms. Simple methanolic extraction proved most effective among tested methods. Separation was achieved on a C12 column with a methanol-ammonium formate mobile phase in five minutes. A dual chemiluminescence system using acidic potassium permanganate and tris(2,2′-bipyridyl)ruthenium(II) provided better detectability than UV absorption at 269 nm, with detection limits of 1.2 × 10⁻⁸ mol/L for psilocin and 3.5 × 10⁻⁹ mol/L for psilocybin. The procedure was applied to three Australian mushroom species.
A new method using flow injection analysis with acidic potassium permanganate and tris(2,2'-bipyridyl)ruthenium(II) chemiluminescence can detect psilocin and psilocybin at very low concentrations—limits of detection are 9×10⁻¹⁰ M for psilocin and 3×10⁻¹⁰ M for psilocybin. The technique is simple, rapid, and sensitive. The paper also describes a concise three-step synthetic route for psilocin from readily available starting materials, with structures confirmed by spectroscopic data.
Aqueous solutions of psilocin and psilocybin reference standards remain stable for up to seven days when protected from light, as measured by high-performance liquid chromatography. Excluding light significantly prolongs their useful life. The study used a methanol and ammonium formate mobile phase with detection at 269 nm over a fourteen-day period.