Development, validation, and application of an ultra-high performance liquid chromatography-tandem mass spectrometry method for the estimation of Ropanicant in human plasma and urine

Abstract

SUVN-911 (Ropanicant) is a novel alpha-4 beta-2 nicotinic acetylcholine receptor (α4β2 nAChR) antagonist being developed for the treatment of major depressive disorders (MDD). An Ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated for the quantification of SUVN-911 in human plasma and urine. Liquid-liquid extraction technique was used to extract the SUVN-911 from human plasma and urine. The separation was performed on an Acquity UPLC® BEH C18 (2.1 × 50 mm, 1.7 µm) column with gradient elution using a mobile phase consisting of 0.01M ammonium acetate and acetonitrile. The QTRAP mass spectrometer which is operated in a positive ionization mode utilizing selective reaction monitoring (SRM) mode has been used for the quantification. The developed method was linear across the standard curve concentrations ranging from 0.020 to 50 ng mL−1 in human plasma and 5 to 5000 ng mL−1 in human urine. The accuracy and precision results were well within the acceptance criteria for experiments performed on different days. The average extraction recovery for plasma and urine were 73.9% (% CV 10.1), and 42.7% (% CV 4.9), respectively, and the average matrix factor and internal standard (IS) normalized mean matrix factor were within the acceptable range of 0.9–1.1. The stability of SUVN-911 in blood, plasma, and urine was established at different storage conditions. The developed and validated UHPLC-MS/MS method was utilized for the quantification of SUVN-911 in plasma and urine samples from the first-in-human (FIH) study.