Trapping and Detection of the Reactive Metabolites
Metabolism of compounds can lead to creation of reactive metabolites which will alter cellular function though binding to cellular proteins as well as nucleic acids. Recognition of the formation of metabolites as well as identification of the metabolites can help to predict adverse effects of a drug.
Incubation of test compound with microsomes in the presence of GSH (unlabeled and stable isotope labeled) will “trap” reactive metabolites and these are detected via characteristic isotopic mass splitting using Orbitrap. “Classical” LC-MS/MS detection methodologies (NL=129; Pr=-272) are also available. Complementary trapping agents (cyanide, e.g.) could be used to expand the scope of the offered assay.