Development and validation of a sensitive LC-MS/MS method with electrospray ionization using multiple ions for...

A highly sensitive and specific LC-MS/MS method has been developed and validated for the estimation of torcetrapib (TTB) with 100 microL hamster/dog plasma using DRL-16126 as an internal standard (IS). The API-4000 Q Trap LC-MS/MS was operated under multiple-reaction monitoring mode using the electrospray ionization technique. The assay procedure involved extraction of TTB and IS from plasma with acetonitrile, which yielded consistent recoveries of 65.73 and 94.01% for TTB and 79.68 and 90.70% for IS in hamster and dog plasma, respectively. The total chromatographic run time was 3.0 min and the elution of TTB and IS occurred at approximately 2.25 and 2.20 min, respectively. The resolution of peaks was achieved with 0.01 m ammonium acetate:acetonitrile (15:85, v/v) at a flow rate of 0.40 mL/min on an Inertsil ODS-3 column. The method was proved to be accurate and precise at linearity range of 1.00-200 ng/mL with a correlation coefficient (r) of > or = 0.993. The method was rugged with 1.00 ng/mL as the lower limit of quantitation. TTB was stable in the battery of stability studies. The application of the assay to preclinical pharmacokinetic studies confirmed the utility of the assay to derive hamster/dog pharmacokinetic parameters.