Highly sensitive determination of TS-962 (HL-004), a novel acyl-CoA : cholesterol acyltransferase inhibitor, in rat and rabbit plasma by liquid chromatography and atmospheric pressure chemical ionization-tandem mass spectrometry combined with a colum

摘要

A quantitative bioanalytical method with excellent specificity using Liquid chromatography (LC) atmospheric pressure chemical ionization-tandem mass spectrometry (APCI-MS-MS) combined with a column-switching technique has been developed for the highly sensitive and reliable determination of TS-962 (HL-004), a novel acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor, in rat and rabbit plasma. The method involves protein precipitation of a 25-mul aliquot of plasma sample with eight volumes of methanol containing a deuterium-labeled internal standard, the direct injection of a methanolic supernatant into the analytical instrumentation with no sample evaporation and reconstitution steps, automated on-line clean-up on a C-18 short trapping column (10 mmx4.0 mm I.D.) followed by separation on a C-18 analytical column (56 mmx4.6 mm I.D.), and detection with APCI-MS-MS using m/z 448 ([M+H](+)) as a precursor ion and m/z 178 as a product ion in a selected reaction monitoring mode. The lower limit of quantification was 1 ng/ml, and good linearity of the calibration graph was obtained in the range of 1 similar to 490 ng/ml with excellent reliability. The developed method enabled pharmacokinetic profiles to be determined for rats and rabbits with sequential plasma collection from an individual animal. (C) 2001 Elsevier Science B.V. All rights reserved. [References: 10]