Quantitative analysis of metabolites in the aflatoxin biosynthesis pathway for early warning of aflatoxin contamination by UHPLC-HRMS combined with QAMS

Aflatoxins seriously threaten human health and food safety, and early warning benefits the reasonable use of control measures to reduce aflatoxin contamination. In this study, a novel method for quantifying aflatoxins and their precursors in the aflatoxin biosynthesis pathway was developed by combining ultra-high performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS) with quantitative analysis of multi-components by a single marker (QAMS). The stability of the relative correction factor (RCF) of QAMS was then systematically evaluated. The validation results showed that the relative deviation (RD) between QAMS and the external standard method (ESM) was less than 11.7%, indicating that the established QAMS method could replace ESM without the use of reference standards. This method was successfully employed to compare the time-course changes of metabolites in the aflatoxin biosynthesis pathway of Aspergillus flavus and Aspergillus parasitica. As a result, the precursors of (1'S,5'R)-5'-Hydroxyaverantin (HAVN) and Versicolorin B (VerB) could be used as potential markers for the early warning of aflatoxin contamination. This study provided a quantitative method of aflatoxins and their precursors in the biosynthesis pathway, and may serve as a reference for the extension of quantitative studies on other metabolic pathways.