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Aflatoxins count to the most toxic known mycotoxins and are a threat to food safety especially in regions with a warm and humid climate. Contaminated food reaches consumers globally due to international trade, leading to stringent regulatory limits of aflatoxins in food. While the formation of aflatoxin (AF) B 1  by the filamentous fungus  Aspergillus flavus  is well investigated, less is known about the formation kinetics of its precursors and further aflatoxins. In this study, autoclaved maize kernels were inoculated with  A. flavus  and incubated at 25 °C for up to 10 days. Aflatoxins and precursors were analyzed by a validated UHPLC-MS method. Additional to AFB 1  and AFB 2 , AFM 1  and AFM 2  were detected, confirming the ability of the formation of M-group aflatoxins on cereals by  A. flavus . The measured relative levels of AFB 2 , AFM 1 , and AFM 2  on maize compared to the level of AFB 1  (mean of days 5, 7, and 10 of incubation) were 3.3%, 1.5%, and 0.2%, respectively. The occurrence and kinetics of the measured aflatoxins and their precursors sterigmatocystin, O-methylsterigmatocystin, 11-hydroxy-O-methylsterigmatocystin, aspertoxin, and 11-hydroxyaspertoxin (group 1) as well as of dihydrosterigmatocystin and dihydro-O-methylsterigmatocystin (group 2) supported the so far postulated biosynthetic pathway. Remarkable high levels of O-methylsterigmatocystin and aspertoxin (17.4% and 4.9% compared to AFB 1 ) were found, raising the question about the toxicological relevance of these intermediates. In conclusion, based on the study results, the monitoring of O-methylsterigmatocystin and aspertoxin as well as M-group aflatoxins in food is recommended.

Formation of B- and M-group aflatoxins and precursors by Aspergillus flavus on maize and its implication for food safety