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Idiosyncratic drug reactions have often caused late stage clinical program failure and drug withdrawal. Formation of oxidative and electrophilic reactive metabolites or intermediates is considered a likely cause of idiosyncratic drug reactions. Therefore, it is reasonable to screen drug candidates in the early stage to eliminate compounds likely to produce reactive metabolites from further development.
Reactive metabolites form conjugates with glutathione (GSH), which can be detected by LC-MS/MS using a constant neutral loss scan of 129 Dalton.
In our glutathione conjugation assay, acetaminophen is used as a positive control. A test compound is incubated in human liver microsomes with the presence of glutathione. Glutathione conjugates are detected by LC-MS/MS. The structure of the conjugates can be elucidated by further MS fragmentation experiments.