Validation and application of a liquid chromatography–tandem mass spectrometric method for the determination of SCH 211803 in rat and monkey plasma using automated 96-well protein precipitation

Abstract

A rapid, sensitive, specific, accurate, and reproducible automated liquid chromatography–tandem mass spectrometric (LC–MS/MS) method for the quantitative determination of 1′-(2-amino-3-methylbenzoyl)-4-[[[(3-chlorophenyl)sulfonyl]phenyl]methyl]-1,4′-bipiperidine hydrochloride (SCH 211803) in plasma has been developed. The method was validated in rat and monkey plasma over the concentration range of 0.5–250 ng/ml using 2 H 4 -SCH 211803 as the internal standard (IS). Automated 96-well plate protein precipitation (PP) with acetonitrile (ACN) was used for sample processing. The method employed a Betasil C18 column with a fast gradient for the separation of analyte and internal standard from the plasma matrix and a triple quadrupole mass spectrometer operated in positive ion multiple reaction monitoring (MRM) mode for detection. The method was used for the determination of SCH 211803 plasma concentrations to support pre-clinical studies.