Quantitative analysis of D‐24851, a novel anticancer agent, in human plasma and urine by liquid chromatography coupled with tandem mass spectrometry

The development of a liquid chromatography/tandem mass spectrometric assay for the quantitative analysis of the novel tubulin inhibitor D-24851 in human plasma and urine is described. D-24851 and the deuterated internal standard were extracted from 250 μL of plasma or urine using hexane/ether (1:1, v/v). Subsequently, 10-μL aliquots of reconstituted extracts were injected onto an Inertsil ODS analytical column (50 × 2.0 mm i.d., 5 μm particle size). An eluent consisting of methanol/5 mM ammonium acetate, 0.004% formic acid in water (80:20, v/v) was pumped at a flow rate of 0.2 mL/min. An API 365 triple quadrupole mass spectrometer was used in the multiple reaction monitoring mode for sensitive detection. For human plasma a dynamic range of 1–1000 ng/mL was validated, and for human urine a range of 0.25–50 ng/mL. Validation was performed according to the most recent FDA guidelines and all results were within requirements. The assay has been successfully applied to support a phase I clinical trial with orally administered D-24851. Copyright © 2004 John Wiley & Sons, Ltd.