Liquid chromatography–mass spectrometry in in vitro drug metabolite screening

Driving efficiency in a high-throughput metabolic stability assay through a generic high-resolution accurate mass method and automated data mining

“…DMPK study for early drug discovery involves various automated in vitro assays such as metabolic stability, cytochrome P450 (CYP) inhibition and induction, and cell permeability study (Di et al, 2009). Metabolic stability is regarded as the most important frontline screen as it plays an important role in determining the overall pharmacokinetic properties of a new drug substance (Tolonen et al, 2009). The metabolic properties of parent drugs as well as their relevant metabolites generated during microsomal incubation provide essential clues to pinpoint promising drug candidates and address their potential issues.…”

“…The metabolic properties of parent drugs as well as their relevant metabolites generated during microsomal incubation provide essential clues to pinpoint promising drug candidates and address their potential issues. As hundreds of discrete compounds need to be screened each week, improving the throughput of metabolic stability assays in a cost-effective manner is highly desired in early DMPK projects (Chu and Nomeir, 2006;Tolonen et al, 2009).…”

“…U-HPLC is typically connected with a triple quadrupole (QqQ) mass spectrometer (MS) or hybrid triplequadrupole linear ion trap (QqQ LIT ) MS so as to monitor the quantitative changes of parent drug concentrations by selected-reaction-monitoring (SRM) analysis. Despite of being a specific and sensitive approach for bioanalysis, SRM approach is laborious and time-consuming because SRM parameters have to be optimized for each compound in the queue and there are hundreds of them that need to be assayed on a weekly basis in a regular DMPK laboratory (Chu and Nomeir, 2006;Tolonen et al, 2009). An additional drawback of QqQ-based targeted analysis is that no data on the drug metabolites can be acquired unless a limited number of known metabolites were predicted prior to the experiment.…”

“…High resolution mass spectrometry (HRMS) has emerged as an attractive approach for early-phase DMPK applications (Tolonen et al, 2009). This approach relies on high resolution and high mass accuracy in full MS scans to derive or confirm the empirical formula of analytes within a wide mass range.…”

Analytical Methods for Quantification of Drug Metabolites in Biological Samples

“…A short overview of analytical strategies for identification of metabolites will be provided. More information regarding metabolite identification can be found in following review articles [7,[23][24][25][26][27]. The selection of suitable LC-MS instrumentation is needed for qualitative evaluation of metabolites.…”