Development and Validation of a Novel High-Performance Liquid Chromatography Assay with Ultraviolet Detection for...

A rapid, simple and highly sensitive stability-indicating high-performance liquid chromatography (HPLC) method was developed and validated for the estimation of asciminib in human plasma. Plasma samples (100 μL) were processed by solid-phase extraction. Chromatographic separation was performed using a CAPCELL PAK C18 MGII column (4.6 × 250 mm, 5 μm); the mobile phase consisted of 0.5% potassium dihydrogen phosphate (KH2PO4, pH 3.5): acetonitrile: methanol (50:45:5, v/v/v) at a flow rate of 0.5 mL/min. Detection was carried out at a wavelength of 250 nm. The developed method showed good linearity over the concentration range of 10-5000 ng/mL (correlation coefficient: 0.9999). The method was then validated according to guidelines published by the US Food and Drug Administration. The coefficients of variation for intra- and interday assays for asciminib were less than 13.9%, whereas accuracies for intra- and interday assays were within 8.9%. The limit of quantification was 10 ng/mL. The mean (minimum - maximum) plasma trough concentration of asciminib in 29 Japanese patients with chronic myeloid leukemia who received repeated administration of 40 mg twice daily was 367 (101-807) ng/mL. The validated method can be used as a routine method to support pharmacokinetic studies of asciminib.