DEVELOPMENT OF ANALYTICAL SPECIFICATIONS AND VALIDATION OF RUTOSIDE TRIHYDRATE USING RP-HPLC

Background: Rutoside Trihydrate is a flavonoid glycoside with vasoprotective and antioxidant properties. The present study aimed to develop and validate an RP-HPLC method for the quantitative estimation of Rutoside Trihydrate in pharmaceutical formulations. Materials and Methods: Rutoside Trihydrate was analyzed using an RP-HPLC system with a UV detector and C18 column (250 × 4.6 mm, 5 µm). Methanol:water (70:30 v/v) was used as the mobile phase at a flow rate of 1.0 mL/min. The injection volume was 20 µL and detection was carried out at 257 nm. The method was validated for system suitability, specificity, linearity, accuracy, precision, sensitivity, robustness, and forced degradation. Results: Rutoside Trihydrate showed a retention time of 4.26 min, with a theoretical plate count of 4862 and a tailing factor of 1.12. The method showed good linearity over 10–50 µg/mL with a correlation coefficient of 0.9997. The mean recovery was 99.90%. The limit of detection and limit of quantification were 0.0063 and 0.0191 µg/mL, respectively. Forced degradation showed 8.42% degradation under acidic, 11.36% under alkaline, 7.85% under oxidative, 5.26% under thermal, and 6.74% under photolytic conditions. The degradation products were separated from the main drug peak. Conclusion: The developed RP-HPLC method was found to be accurate, precise, sensitive, robust, and stability-indicating. The assay of the pharmaceutical formulation was 99.42% with a %RSD of 0.09%. The method is suitable for routine estimation and quality control of Rutoside Trihydrate in pharmaceutical formulations.