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.
Recommended ResourcesApplication of Glutathione (GSH) and Stable Isotope Labeled GSH for Trapping Reactive Metabolites in Cryopreserved and Fresh Hepatocytes Followed by Analysis Using High Resolution Mass Spectrometry (HRMS)
Effects of prototypical microsomal enzyme inducers on cytochrome P450 expression in cultured human hepatocytes
SafetySuite Metabolism and Transporter Package