Bioanalytical Method Development and Validation of Selinexor in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry

Pharmaceutical Science-Pharmaceutical Analysis

Abstract

A simple, rapid, precise, sensitive, and reproducible reverse-phase high-performance liquid chromatography (RPHPLC) LCMS/MS method has been developed for the bioanalytical method for Selinexor with D6-Selinexor as Internal Standard in the pharmaceutical dosage form. Chromatographic separation of Selinexor was achieved on Waters Alliance-e2695, by using X-Bridge phenyl, 150x4.6mm, 3.5μm column, and the mobile phase containing 0.1% Formic acid & Acetonitrile in the ratio of 80:20% v/v. The flow rate was 1.0 ml/min; detection was carried out by absorption at 225nm using a photodiode array detector at ambient temperature. The method was validated to fulfill International Conference on Harmonization (ICH) requirements and this validation included specificity, selectivity, matrix effect, linearity, the limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy. The proposed method was Bio-analytical validated according to USFDA guidelines. This method was found to be a very simple, economical, suitable, precise, accurate, and stable method for pharmacokinetic analysis of Selinexor and study of its stability. The calibration curve was linear over the concentration range from 0 to 40 ng/ml, and the lower limit of detection of 12.5 ng/ml. The accuracy and precision of the method were within the acceptable limit of ±20% at the lower limit of quantitation and ±15% at other concentrations. Selinexor was unstable at room temperature it showed more than 25% loss after 24 h. While, Selinexor is very stable at refrigerator 40C auto-sampler, freeze/thaw cycles, and 30 days storage in a freezer at 35 ± 20C. All results were acceptable and this confirmed that the method is suitable for its intended use in routine quality control and an assay of drugs.