Purification of High Purity Alpha Amanitin Using Preparative HPLC Method

Objective: Alpha-amanitin is purified only around 90% purity using existing methods. In this study, it was aimed to describe the method in order to obtain high-purity alpha-amanitin using preparative HPLC. Methods: Amanita phalloides mushroom was collected, extracted and purified 2 times using preparative HPLC. Validation of the toxin was performed by comparison of retention time and ultraviolet spectrum at HPLC. Results: Alpha-amanitin was obtained with 93% (+/- 1.24) purity after first purification process. Alpha-amanitin was obtained with 99,8% (+/- 0.26) purity after second purification process. It was seemed that purified toxin and standard were given maximum absorbance at 303 nm and minimum absorbance at 263 nm, and the structure of the spectrums for both was similar. Conclusion: Alpha-amanitin with >99% purity can be obtained by this method at low cost.