Determination, Purification And Chemical Structure Identification Of Main Peptide Toxins From Amanitas

The determinnation, purification and chemical structure of amatoxins from Amanita were studied, the results are follow:1. The analysis of main peptide toxin from 28 species amanita which collected from China and A. phalloides collectedxfrom Germany showed that Amanita fulliginea, A. exitialis, A. subjunquillea S. Imai var. alba, endemic to East Asia, and A. phalloides contained manykinds of toxic peptides and display a very high amount of toxins. The total content of peptide toxins are 15327. 5 μ g/g, 7780. 3μ g/g, 1013. 9μ g/g and 7632. 4μ g/g dried weight carpophore respectively. These four amanitas may regard as lathel amanitas. There are no any amatoxins of stardard samples in other fifteen species, and there are trace amatoxins in other ten species and the contents were about 19. 5μ g/g~151. 2 11 g/g dried fruitbody.2. The content and distribution of the main amanitoxins and phallotoxins in three tissues (cap, stipe and volva) of the four lethel amanitas were evaluated. The result showed that the highest content of total tcxins was displayed in the pileus. The total toxins content in stipe was lower, whereas the volva had the lowest content of total toxins. The content of amanitoxins is always higher than the content of phallotoxins in any tissues of the four amanitas but A.phalloides. Furthermore, the relative content of phallotoxins isinceasing from cap to stipe to volvain in the four lathel amanitas.3. The total amatoxins concentration from carpophore of A. fuliginea collected in different development stages were studied. The result showed that the highest content of total amatoxins (23793. 3 μ g/g) was found in the carpophore with the cap free from veil but the gill concealed by veil and, the lowest content of total amatoxins (4674. 9μ g/g) was in the carpophore which has withered. The content of amatoxins decent rapidly when it is withering.4. The total amatoxins from A. fuliginea carpophore in different size were studied. The result showed that when the diameter of the carpophore no more than 5.0cm, the bigger the carpophore is, the more main amatoxins content is, but when diameter of the carpophore pileus reach 7.0cm, the total amatoxins content decent in some degree.5. Isolation and purification of four main peptide toxins from A. fuligiea were undergone by means of RP-HPLC, and idenfication of the chemical structure of α-amanitin and β-amanitin by nuclear magnetic resonance (NMR) showed that the the chemical structure were the same as the reference.