High-resolution mass spectrometry-based suspect and nontarget screening of natural toxins in foodstuffs and risk assessment of dietary exposure

Daily dietary intake inevitably exposes individuals to various natural toxins, which may pose potential health threats. Focusing only on specific toxins could underestimate dietary risks. Therefore, we have developed a suspect and nontarget method based on ultrahigh-performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) to screen both known and unknown natural toxins in various foodstuffs. An in-house database containing 2952 natural toxins including fungal toxins, phytotoxins, animal toxins and cyanotoxins was established, facilitating suspect screening. Predicted retention time and mass spectrometry data were employed to enhance the confidence levels. Subsequently, Nontarget screening method was conducted based on molecular network analysis, annotating the edges and nodes through modified types and fragmentation characteristics. Finally, we analyzed 102 foodstuff samples and identified a total of 90 natural toxins, including mycotoxins and phytotoxins, with 65 identified by suspect screening and 25 by nontarget screening. Based on measured concentrations, the daily per capita dietary intake of total natural toxins was estimated, it was below risk doses for natural toxins with available reference values. Overall, this work established a novel method for the comprehensive identification of natural toxins in foodstuffs and emphasized the importance of dietary risk assessment for natural toxins.