The present invention discloses a method for the isolation and purification of amatoxins in poisonous mushrooms. The specific steps include: S1. Firstly, separating the
mushroom extract containing amatoxins by high-speed counter-current
chromatography; S2. Secondly, separating the product after the first separation by preparative liquid
chromatography. The present invention uses HSCCC combined with Pre-HPLC to isolate and purify amatoxins in
Amanita mushrooms, collects the target substances in groups, and finally conducts purity analysis and structural confirmation, thereby establishing a method for isolating and purifying four components, namely α-AMA, β-AMA, carboxy-tricholoro-phalloidin, and carboxy-dihydroxy-phalloidin, in
Amanita mushrooms. This method is simple to operate, fast and efficient, with high purity, good reproducibility, and high sample
recovery rate.