Related substances method development and validation of an LCMS/ MS method for quantification of selexipag and its related impurities in rat plasma and its application to pharmacokinetic studies

摘要

The present application wish to seem at the event of validation of bio analytical method and pharmacokinetic study ofselexipag and its related impurities in rat plasma using LC–MS/MS. The optimized method contains gradient elution ofselexipag with a flow rate of 1 ml/min and X-Bridge phenyl column (150 × 4.6 mm, 3.5 μ). A buffer of 1 mL formic acid inl liter water and acetonitrile mixture is used as mobile phase. 30 min run time was used for separation of selexipag andits related impurities with Ambrisentan as internal standard and impurity-D as active metabolite. The linearity curves arelinear in between the percentages of 10 to 200% of rat plasma and R~2 value of each analyte was observed as 0.999. Thisapplication denotes all the parameters like precision, accuracy, recovery and stability were got the results within the limitof USFDA guidelines. This method applies effectively for the investigation of pharmacokinetic studies using rat plasma.