HPLC Analytical Research Method Development and Validation of Inosine Pranobex by using the QbD Approach.

The study highlights the development of a high-performance liquid chromatographic (HPLC) method for estimating Inosine Pranobex using a Quality by Design (QbD) approach. This risk-based method development emphasizes risk assessment and management to achieve robustness. A 2-factorial experimental design, utilizing a central composite design, was used to optimize the mobile phase and flow rate, aided by Design Expert software. Optimal conditions included a C18 column (4.6 x 100 mm), a mobile phase of methanol to 0.1% OPA in a 60:40 ratio, and a flow rate of 1 ml/min, resulting in a retention time of 6.1 minutes. The method showed linearity (r2 = 0.999) in the 10–50 μg/ml range at a detection wavelength of 259 nm. Intraday and interday precision %RSD values were between 0.40–1.5 and 0.0–0.61, respectively. Robustness values were under 2%, and the assay result was 99.97 ± 0.03%. The chromatographic peak purity analysis confirmed the absence of co-eluting peaks with Inosine Pranobex. The method validation complied with ICH guidelines, demonstrating a successful HPLC method development with an improved understanding of performance-related variables.