Abstract
<title>Abstract</title> <p>Background Desidustat, a per-oral hypoxia-inducible factor prolyl hydroxylase inhibitor sanctioned for erythropoietic deficiency in chronic kidney disease, currently lacks a rigorously validated, design-guided chromatographic assay procedure. This investigation sought to establish and validate a systematic, A QBD-directed RP-HPLC technique for precise quantitative determination of desidustat in both bulk pharmaceutical material and solid oral tablets, addressing the need for reliable industrial quality surveillance and prospective clinical pharmacokinetic applications. Methods Physicochemical characterization encompassing UV spectrophotometric profiling, thermal melting point analysis, and aqueous solubility assessment was conducted to examine intrinsic drug substance attributes. A 3² full-factorial experimental matrix was implemented to rationally optimize oven temperature (30–50°C) and mobile phase flow velocity (0.8–1.2 mL/min) as independent input parameters, while peak retention time and tailing asymmetry constituted measurable chromatographic responses. Baseline resolution was accomplished using an Inertsil ODS-3V C18 stationary phase (150 × 4.6 mm, 5 µm), employing an isocratic Ethanol:Water binary eluent (70:30 v/v) with UV detection at 229 nm. Procedural performance attributes were fully verified per ICH Q2(R2) regulatory guidance. Results Composite desirability modeling pinpointed Run 8 conditions (50°C; 1.0 mL/min) as the globally superior setting, attaining a maximum desirability index of 1.0. The certified procedure exhibited outstanding linearity spanning 5–15 µg/mL (r² = 0.9997), trueness of 99.93 ± 0.26%, intra-day repeatability %RSD of 0.22%, inter-day intermediate precision %RSD of 0.16%, LOD of 0.14 µg/mL, and LOQ of 0.43 µg/mL. Enforced stress degradation investigations corroborated stability-indicating performance, with mass balance recovery surpassing 98%, while commercial tablet quantification returned 99.95 ± 0.22% of labeled potency. Conclusion The A QBD-engineered RP-HPLC procedure satisfies all ICH Q2(R2) specifications, exhibiting specificity, sensitivity, accuracy, precision, robustness, and stability-indicating character. Adoption of an ethanol-based mobile system upholds sustainable green analytical chemistry tenets, constituting a dependable quantitative platform for industrial quality oversight of desidustat and offering clear applicability to future bioequivalence, pharmacokinetic, and translational clinical research.</p>