Tentative identification of the composition of Agaricus bisporus aqueous enzymatic extracts with antiviral activity against HCV: A study by liquid chromatography–tandem mass spectrometry in high resolution mode

• Sequential fractionation of AbAEE with anti-VCH protease NS3 activity.
• Use of different polarity extractants for efficient AbAEE fractionation.
• 123 compounds were tentatively identified by MS/MS information.
• 55 out of the 123 compounds tentatively identified had not been identified before.
• LC–QTOF MS/MS was confirmed as strategy for overall metabolome coverage of AbAEE.

Aqueous enzymatic extracts of edible mushroom Agaricus bisporus (AbAEE) have been studied as a key source of biological active compounds, among which those with antiviral activity against HCV are of special interest. Searching for a better knowledge of AbAEE, a new approach based on sequential fractionation with solvents of different polarity is here proposed to improve the tentative identification of AbAEE components by liquid chromatography–tandem mass spectrometry (LC–QTOF MS/MS) in high resolution mode. Fifty five out of one hundred twenty-three total metabolites tentatively identified by combination of MS and MS/MS information were identified for the first time. The newly identified compounds include amino acids, sugars, carboxylic, fatty and cinnamic acids, mono- and disaccharides, phospholipids, and purines as the most outstanding, thus demonstrating that fractionation based on sequential liquid–liquid extraction followed by LC–QTOF MS/MS is a suitable option to obtain a wide, representative snapshot of AbAEE composition.