QbD-Driven Development and Validation of a Novel RP-HPLC Method for Quantitative Determination of Coenzyme Q10 in Single and Combined Dosage Forms

The objective of the study was to develop and validate an authentic RP-HPLC methods based on Quality by Design (QbD) for the quantitative analysis of Coenzyme Q10 (CoQ10) in both single and combined dosage forms with clomiphene citrate (CC). Risk assessment, Design of Experiments (DoE), and the creation of the Method Operable Design Region (MODR) were all part of a methodical QbD approach. Screening tests (n=18) improved flow rate, mobile phase composition, and column chemistry. The impact of flow rate and percentage CAN on Critical Quality Attributes (CQAs), including Retention Time (RT), Resolution (Rs), and Tailing Factor, was investigated using a DoE (32 factorial design). Under ideal chromatographic conditions, a C18 column (250 × 4.6 mm, 5 µm), 70% CAN, phosphate buffer (pH 3.0), and a flow rate of 1.0 mL/min with UV detection at 275 nm were used. The process was confirmed to satisfy ICH Q2 (R1) requirements. 86 minutes for CoQ10 and approximately 4.05 minutes for CC. Acceptance criteria were met by the system appropriateness metrics (Resolution >35, Tailing ~1.3, Theoretical Plates >30,000). Great linearity was demonstrated by the method (2.5–200 µm/mL; R2=0.9997 CoQ10, R2=0.9996 CC). Research on accuracy revealed recoveries at 80%, 100%, and 120% levels that fell between 98 and 102%. Precision was confirmed by intra- and inter-day %RSD being less than 2%. Minimal differences in system appropriateness with intentional changes in flow rate and %ACN were indicative of robustness. After 24 hours at room temperature and 48 hours in the refrigerator, the solutions held their stability. In comparison to previously published methods, its QbD–driven approach guarantees regulatory compliance and superior method performance.