Thermostability of Alpha Amanitin in Water and Methanol

Alpha amanitin mostly found toxin in Amanita phalloides mushroom is responsible for toxicity. In this study, the chemical stability of the alpha amanitin in water and methanol at different temperatures was investigated. Alpha amanitin was purified by the method mentioned earlier from Amanita phalloides extract, and compared to the standard. Obtained toxin was dissolved in distilled water and methanol, and separated into 42 vials. Vials divided into 7 groups randomly. The stability of the toxin was measured at room temperature (25 °C), in the freezer (-20 °C), refrigerated (4 °C) and boiling (only water) (98 °C) using analytical HPLC system. In this method, 4.6 x50mm reverse phase C18 column, UV detector at a wavelength of 303nm, 1 mL/min flow rate of mobile phase (acetonitrile ammonium acetate + methanol + 80 +10 +10) were used. The initial amount of toxin was taken as100%, and toxin ratio was calculated according to the time, reduction in the amount of toxin. The remaining toxin level was only 5% after 6 hours in boiled water. The remaining toxin level was 86%, and 96% after 6 mounts in water, and methanol respectively at room temperature. The remaining toxin level was 91%, and 97% after 6 mounts in water, and methanol respectively at 4 °C. The remaining toxin level was 76%, and 97% after 6 mounts in water, and methanol respectively at -20 °C. Almost all of the toxin with boiling 6 hours changed to another chemical structure, but this does not imply toxicity completely disappeared. The most stability in the water was measured at 4 °C. However, the most stability in the methanol was measured at all temperatures. The chemical degradation does not mean loss of toxicity. To clarify it needs further invesitagation.