Quantitative determination of a selective alpha-1a receptor antagonist in human plasma by high-performance liquid chromatography with tandem mass spectrometric detection

Solid-phase extraction, utilizing a 96-well plate format, was used to isolate an alpha-1a receptor antagonist and internal standard from human plasma. Following the isolation procedure, the analyte and internal standard were separated and detected using reversed-phase HPLC coupled with atmospheric pressure chemical ionization (APCI) mass spectrometry operated in the positive ion multiple reaction monitoring (MRM) mode. Based upon the peak area ratio (analyte: internal standard) the analyte was quantified over a concentration range of 0.02–2 ng/ml. Assay validation results including parameters such as precision and accuracy are presented. The validated method was subsequently used to support human pharmacokinetic studies.