Tandem mass spectrometry imaging to assess illudin S distributions in Omphalotus illudens mushrooms

Poisoning metabolites within mushrooms typically exert varying physiological and biological effects; however, their distributions within mushrooms are seldom investigated. This study focused on imaging illudin S, a representative poisoning metabolite within Omphalotus illudens mushroom, using desorption electrospray ionization-tandem mass spectrometry (DESI-MS/MS) coupled with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Initially, LC-ESI-MS/MS conditions were established for the selective detection of illudin S (m/z 247 -> 159, positive mode) from extracted mushroom. Based on this MS/ MS condition, each mushroom was divided into nine sections, and each section was subjected to specific DESI-MS/MS mode imaging (pixel size: 200 x 200 mu m). The results revealed high concentrations of illudin S in the stalks of the mushrooms growing on trees. Further, re-evaluations using the LC-MS/MS assay revealed a seven-fold difference in the illudin S between the stalk and other regions. This study marks the first attempt at assessing toxic metabolite distributions within poisonous mushrooms, thus offering valuable insights including the efficient utilization of illudin analogs in biological applications.