High resolution mass spectrometry workflow for the analysis of food contaminants: Application to plant toxins, mycotoxins and phytoestrogens in plant-based ingredients

ABSTRACT

An analytical workflow including mass spectral library, generic sample preparation, chromatographic separation, and analysis by high-resolution mass spectrometry (HRMS) was developed to gain insight into the occurrence of plant toxins, mycotoxins and phytoestrogens in plant-based food. This workflow was applied to 156 compounds including 90 plant toxins (pyrrolizidine alkaloids, tropane alkaloids, glycoalkaloids, isoquinoline alkaloids and aristolochic acids), 54 mycotoxins (including ergot alkaloids and Alternaria toxins) and 12 phytoestrogens (including isoflavones, lignans and coumestan) in plant-based protein ingredients, cereal and pseudo-cereal products. A mass spectral library was built based on fragmentation spectra collected at 10 different collision energies in both positive and negative ionisation modes for each toxin. Emphasis was put on a generic QuEChERS-like sample preparation followed by ultra-high-pressure liquid chromatography using alkaline mobile phase allowing the separation of more than 50 toxic pyrrolizidine alkaloids. HRMS acquisition comprised a full-scan event for toxins detection followed by data-dependent MS2 for toxin identification against mass spectrum. Method performance was evaluated using fortified samples in terms of sensitivity, repeatability, reproducibility and recovery. All toxins were positively identified at levels ranging from 1 µg kg⁻¹ to 100 µg kg⁻¹. Quantitative results obtained by a standard addition approach met SANTE/12682/2019 criteria for 132 out of 156 toxins. Such a workflow using generic, sensitive and selective multi-residue method allows a better insight into the occurrence of regulated and non-regulated toxins in plant-based foods and to conduct safety evaluation and risk assessments when needed.

Graphical Abstract

KEYWORDS:

• Natural toxins
• plant toxins
• mycotoxins
• pyrrolizidine alkaloids
• tropane alkaloids
• high-resolution mass spectrometry

Acknowledgments

The authors would like to thank Dr. Thierry Delatour and M. Pascal Mottier for their valuable input in the preparation of this manuscript.

Credit author statement

Thomas Bessaire: Methodology, Software, Formal analysis, Investigation, Writing – Original Draft.

Marion Ernest: Methodology, Software, Formal analysis, Investigation, Writing – Original Draft.

Nicolas Christinat: Methodology, Software, Formal analysis, Data Curation, Investigation, Writing – Original Draft.

Benoit Carrères: Software, Writing – review & editing.

Alexandre Panchaud: Conceptualisation, Resources, Supervision, Writing – review & editing.

Flavia Badoudy: Conceptualisation, Methodology, Software, Formal analysis, Investigation, Writing – Original Draft, Supervision, Project administration.

Supplementary material

Supplemental data for this article can be accessed on the publisher’s website.