SIMULTANEOUS ESTIMATION OF RUTIN USING HPLC METHOD FROM AYURVEDIC FORMULATION

Abstract

A precise and reliable High-Performance Liquid Chromatography (HPLC) method was developed and validated for the simultaneous estimation of Rutin in Ayurvedic formulations, including syrup and tablet dosage forms. Rutin was used as a reference standard, and standard stock and working solutions were prepared using methanol. A calibration curve was constructed across a concentration range of 6–18 µg/mL, demonstrating excellent linearity with a correlation coefficient (r²) of 0.9985. The mobile phase, optimized for sharp peak resolution, consisted of 70% methanol and acetate buffer (pH 4.5) in a 95:5 (v/v) ratio. Sample solutions were prepared by ultrasonic extraction followed by filtration. The method was validated in accordance with ICH Q2(R1) guidelines. Specificity was confirmed by the absence of interference at Rutin's retention time. Precision studies, including system precision, method precision, and intermediate precision, showed %RSD values below 2%, indicating high reproducibility. The LOD and LOQ were determined to be 0.321 µg/mL and 0.921 µg/mL, respectively. Accuracy was established through recovery studies at 80%, 100%, and 120% levels, with mean recoveries ranging from 99.35% to 100.73%. Robustness testing under varied flow rates, detection wavelengths, and column temperatures demonstrated the method's stability and reliability. This validated HPLC method is suitable for routine quality control of herbal and Ayurvedic formulations containing Rutin, ensuring both accuracy and precision in quantification.