Nicotine in the button mushroom Agaricus bisporus, endogenous biosynthesis?

In early 2009 nicotine was unexpectedly detected in dried mushroom samples. As its origin has not yet been elucidated, this study addressed possible endogenous synthesis of nicotine. Therefore, Agaricus bisporus fruiting bodies were grown in a representative and controlled (nicotine-free) setup. Fruiting bodies (fresh versus stored, intact versus processed (sliced/cooked)) from different harvest days and flushes were analysed with a validated, sensitive dilute-and-shoot UHPLC-MS/MS methodology for nicotine and its precursors putrescine and nicotinic acid. Neither storage nor processing initiated any endogenous nicotine biosynthesis (detection limit 1.6 ng g^-1 fresh weight). In contrast, putrescine and nicotinic acid were detected in all samples, with increasing amounts in the different treatments. In silico analysis of the fully sequenced genome of A. bisporus confirmed its inability to produce nicotine. The data obtained do not provide evidence for natural, endogenous presence of nicotine in mushrooms, indicating an exogenous contamination source (e.g. contamination during hand-picking, sample preparation/analysis).