Ultra-fast quantitative bioanalysis of a pharmaceutical compound using liquid chromatography-tandem mass spectrometry

Abstract

This paper describes the ultra-fast determination of a pharmaceutical compound using TurboIonSpray™ LC-MS-MS on an API 4000 mass spectrometer. Sample preparation consisted of plasma protein precipitation, centrifugation and dilution of the supernatant. The use of small analytical column dimensions ( 2.1 mm×10 mm) and high eluent flow rates (up to 2.2 ml/min) in isocratic mode led to a retention time of 9 s. A sample-to-sample cycle time of only 10 s was achieved by coupling two autosamplers. Partial separation of the drug and its main metabolite could be obtained. The d5-labeled drug used as internal standard compensated for matrix suppression effects. The assay was linear in the concentration range 1–1000 ng/ml, using standards prepared in human plasma. Inter-assay accuracy and precision were 98.5 and 6.2%, respectively. Mean intra-assay accuracy and precision calculated from quality control (QC) samples in human, rat and dog plasma at 3, 30 and 800 ng/ml were 100.8 and 3.8%, respectively. The ultra-fast LC-MS-MS method was successfully cross-validated against a commonly used column-switching LC-MS-MS assay with 2.3 min run time by analyzing real study samples.