Occurrence of psilocybin in various higher fungi from several European countries.

Using high performance liquid and thin-layer chromatographic methods more than 100 species of fungi from Europe belonging to 18 genera were analysed for the hallucinogenic compound psilocybin and for some related tryptamine derivatives. Psilocybin, psilocin and/or baeocystin were found in 3 PSILOCYBE 1 PANAEOLUS, 5 INOCYBES and one PLUTEUS. 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.

Determination of alpha-, beta-, and gamma-amanitin by high performance thin-layer chromatography in Amanita phalloides (Vaill. ex Fr.) secr. from various origin.

A fast, sensitive high performance thin-layer chromatographic method for the determination of alpha-, beta-, and gamma-amanitin in crude, methanolic extracts of Amanita phalloides is described. The limit of detection is 50 ng of each amanitin. With this method amanitin was determined in 24 pooled samples of Amanita phalloides, collected between 1970 and 1977 in Germany and Switzerland. The total amanitin content varied between 2010 and 7300 mg/kg dry weight and the average value was 4430 mg/kg of which 43% was alpha-amanitin, 49% beta-amanitin and 8% gamma-amanitin. The origin of the fungi hardly influenced their amanitin content: in samples collected during the same year at different sites it fluctuated within a factor of 1.7. The amanitin content of samples from the same site, but collected in different years, maximally varied within a factor of 3.7. The partial decomposition of amanitins during prolonged storage of the lyophilized samples undoubtedly contributed to this variation. Phalloidin, which was determined by conventional thin-layer-chromatography, could not be detected in a sample from 1970, whereas its concentration in material collected during 1977 amounted to 2400 mg/kg dry weight. The toxicity of the samples (LD50 of lyophilized defatted methanolic extracts intravenously for mice) varied within a factor of 2.5.

Amanitin content and toxicity of Amantia verna Bull.

The alpha-, beta-, and gamma-amanitin content of 11 samples of Amanita verna Bull., collected during 1975--1978 in Germany and Switzerland, has been determined by high performance thin-layer chromatography (HPTLC) of crude methanolic extracts. The toxicity (i. v. LD50 for mice of defatted, lyophilized, methanolic extracts) of 3 samples has been compared with that of A. phalloides from the same site of collection. The amanitin content of A. verna ranged from 2250 to 4570 mg/kg dry weight; the fungi contained almost as much beta- as alpha-amanitin, whereas the gamma-amanitin content amounted to about 12% of the total amanitin. A verna contained less amanitin (65% on the average) than A. phalloides from the same collection site, but it was not significantly less toxic, since the phallotoxins contributed to the toxicity of either species in our tests.

Selenium content of mushrooms.

The selenium contents of 83 species of wild mushrooms were determined by oxygen combustion of the sample, followed by conversion of selenite to bromopiazselenol and final estimation by electron capture gas-liquid chromatography. Selenium concentration were found to range from 0.012-20.0 mg/kg dry weight. Selenium content was species-dependent. High concentrations were found in Agaricaceae and in certain Boletaceae of the genus Tubiporus, whereas in Russulaceae, Amanitaceae and Cantharellaceae selenium-rich species were absent or rare. Ascomycetes and all mushrooms growing on wood had a very low selenium content. The highest selenium concentrations (up to 20 ppm) were found in Boletus (Tubiporus) edulis, a most popular edible mushroom. Analyses of various parts of carpophores of B. edulis, Suillus luteus and Amanita muscaria indicate that in all three species the stalk contains less selenium than the fleshy part of the cap. In Boletus and Suillus the highest selenium content was found in the tubes.

Rapid determination of selenium in various substrates by electron capture gas-liquid chromatography.

In the proposed procedure for the determination of selenium, 0.5-1 g of sample is wet ashed with concentrated nitric acid. After adding 1,2-diamino-4,5-dichlorobenzene to the digest at pH 1, the resulting dichloropiazselenol derivatives is extracted with toluene. The extract is purified by column chromatography over Florisil and analyzed by gas-liquid chromatography with electron capture detection. Recoveries of selenium added to various substrates ranged from 72 to 102%. The limit of detection is approximately 0.01 ppm, but smaller amounts can be determined by increasing the sample size or by concentration of the final extract.