Amanitin poisonings are among the most life-threatening
mushroom poisonings, and are mainly caused by the genus Amanita. Hepatotoxicity is the hallmark of
amanitins, powerful toxins contained in these mushrooms, and can require
liver transplant. Among amatoxins, α-
amanitin is the most studied. However, the hypothesis of a possible metabolism of
amanitins is still controversial in this pathophysiology. Therefore, there is a need of clarification using cutting-edge tools allowing metabolism study. Molecular network has emerged as powerful tool allowing metabolism study through organization and representation of untargeted tandem mass spectrometry (MS/MS) data in a graphical form. The aim of this study is to investigate
amanitin metabolism using molecular networking. In vivo (four positive
amanitin urine samples) and in vitro (differentiated HepaRG cells supernatant incubated with α-
amanitin 2 μM for 24 h) samples were extracted and analyzed by LC-HRMS/MS using a Q Exactive™ Orbitrap mass spectrometer. Using molecular networking on both in vitro and in vivo, we have demonstrated that α-
amanitin does not undergo metabolism in human. Thus, we provide solid evidence that a possible production of
amanitin metabolites cannot be involved in its toxicity pathways. These findings can help to settle the debate on
amanitin metabolism and toxicity.