LC-MS/MS based bio analysis of desidustat: from development to application

Abstract

Background: Desidustat's potential as a medicinal treatment for anaemia has generated considerable interest. For pharmacokinetic analysis, therapeutic monitoring, and drug development research, precise quantification of Desidustat in biological matrices is crucial. Hence, more advanced methods are necessary for detection at minute concentrations (ng/mL level) in plasma. Methodology: A reverse-phase C18 column with an isocratic elution technique utilising acetonitrile and an aqueous buffer as mobile phase to accomplish chromatographic separation. To produce characteristic ion transitions for quantification, after electrospray ionization (ESI), tandem mass spectroscopy in positive ion mode was chosen for detection. The technique's linearity, sensitivity, precision, and accuracy were systematically improved and verified, and the validated method was successfully applied to pharmacokinetic studies. Results and Discussion: The developed method’s validation results confirmed good selectivity, sensitivity (LLOQ 6 ng/ml), excellent linearity, and acceptable precision and accuracy, with a remarkably low lower limit of quantification (LLOQ). Recovery and matrix effect studies showed minimal ion suppression and reproducible extraction efficiency. The stability of desidustat is demonstrated in rat plasma. Intra-day and inter-day precision and accuracy values were well within the acceptable limits prescribed by regulatory guidelines, confirming the method's reliability and reproducibility. The optimized LC–MS/MS conditions ensured high sensitivity and selectivity, making the method suitable for bioanalytical applications. Conclusion: A novel, highly sensitive, and reliable LC–MS/MS bioanalytical method for estimation of Desidustat in rat plasma was successfully developed and validated. The method provides a robust analytical tool to support pharmacokinetic studies, therapeutic monitoring, and future research related to Desidustat.