Analysis of the toxins of amanitin-containing mushrooms.

We have recently developed a method for the analysis of toxins in single specimens of the mush­ room Amanita phalloides (Fr.) Seer.1. Separation of the different toxins was achieved by column chro­ matography, on Sephadex LH20 in water, of a methanolic extract of fresh or dried mushrooms, followed by further purification on thinlayers of silica gel, spectrophotometric evaluation, and, final­ ly, identification of the toxins by amino acid analy­ ses of their y-hydroxylated leucines or isoleucines as lactones. This procedure afforded the quantitative determination of up to 10 toxins in 2 samples of A. phalloides from different sources. In the present study the treatment outlined above was extended successfully to all the amanitincontaining species of mushrooms known in Europe so far. We examined two more samples of A. phal­ loides collected near Hirschhorn (Neckar, W.-Ger­ many) in 1971 (sample III) and 1972 (sample IV), so that the analytical results from the 4 different samples of this species should shed light on the question as to what extent the toxic composition may differ, depending on location and season. A single small sample of the white species Amanita verna, of only 1.47 g of dry weight, was found near Würz­ burg (W.-Germany) in 19712. Two samples of withe Amanita virosa were collected near Trento, Italy, 1972 (sample I) 3 and in Hägnan, Femsjö (Smoland, Sweden) in 19724 (sample II) . We also investigated a sample of Galerina marginata (Fr.) Kühn, collected near Einsiedeln (Switzerland) in 19724; these small, brown mushrooms, which grow on coniferous trunks, are, together with the stirpes G. automnalis and G. venenata, the only amanitin-containing mushrooms outside the Amanita group known to date. This had already been re-