Applying green analytical chemistry for development and validation of RP-HPLC stability indicating assay method for estimation of fenoverine in bulk and dosage form using quality by design approach

Abstract

ABSTRACTA green and robust reverse-phase liquid chromatographic method has been developed for the determination of fenoverine (FEN), by applying combined principles of green analytical chemistry and quality by design approaches on a Spherisorb C18 column (150 × 4.6 mm, 3 µm) with UV detection at 262 nm. A two level fractional factorial design (2^7-3) Res IV was used for screening of influential chromatographic factors. The critical method parameters actively affecting critical quality attributes (CQAs) were identified and further optimized using Box–Behnken design. The predicted optimum assay conditions comprised of methanol and ammonium acetate buffer 20 mM, in an extent of 81:19% v/v individually having a flow rate of 1.0 mL/min with a column oven temperature of 33°C. The drug was stressed in hydrolytic, oxidative, reductive, thermal, and photolytic conditions. The developed method was validated successfully. The detector response was linear in the concentration of 0.5–160 µg/mL with a limit of detection (LOD) and limit of quantitation (LOQ) as 0.1 and 0.3 µg/mL, respectively. The % recovery was found to be 99.7%. The analytical method volume intensity value for developed method was 45 mL and the environment assessment tool (EAT) score was 41.07. The method is simple, environmentally benign, rapid, and robust for the determination of FEN in bulk and in its dosage form.