A semi-automated method for the integrated evaluation of half-life and metabolic soft spots of discovery compounds. (July 2012)

• Record Type: Journal Article
• Authors: Grubb, Mary F; Humphreys, William G; Josephs, Jonathan L
• Is Part Of: Bioanalysis. Volume 4:Number 14(2012)
• Page Start: 1747
• Page End: 1761
• Publication Date: 2012-07
• Subjects: Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio.12.134
• Languages: English
• ISSNs: 1757-6180

Abstract

Background: An integrated method that provides rates of both parent disappearance and metabolite formation was developed.Results: Buspirone, mirtazapine and verapamil were used as model compounds in developing the method. Incubations were carried out on a robotic platform. Qualitative analysis of metabolites in 30 µM samples was conducted by data-dependent HPLC–MS/MS on a high-resolution instrument. Quantitative analysis of the parent compound and metabolites in 0.5 µM samples was conducted by full-scan MS 2 with product ion extraction using an ion trap mass spectrometer. Data generated for the compounds included half-life and intrinsic clearance of the parent molecule, characterization of metabolites and relative rates of metabolite formation. A correction factor was used to convert MS responses of metabolites in 0.5 µM samples to UV areas in order to compare relative metabolite concentrations.Conclusion: The approach allows for the investigation of a set of six compounds simultaneously, with a turnaround time of 1 week or less.