A Novel Validated Stability Indicating Chromatographic Method for the Simultaneous Estimation of Ascorbic Acid and Gallic Acid in the Ayurvedic Capsule Dosage form of Amla by UFLC

An Ultra Fast liquid chromatographic method has been developed and validated for simultaneous estimation of ascorbic and gallic acid in both API and Ayurvedic formulation. Chromatographic separation of compounds was carried out with C8 column by using a mobile phase of methanol: phosphate buffer, PH 3.0 (1: 5) at a flow rate of 1.0 ml/ min. UV detection was performed using PDA detectors at 264 nm. The method was validated for accuracy, precision, linearity, LOD, LOQ and robustness in accordance with ICH guidelines. Amounts of ascorbic and gallic acid detected in capsule were 99.20% and 99.45%. Total run time was below 3 min, ascorbic and gallic acid was eluted with retention times of 1.541 and 2.591 min respectively. Validation revealed that the method is specific, accurate, precise, reliable and reproducible. Calibration plots were linear over the concentration ranges 1-9 μg/ ml for ascorbic acid and for gallic acid, respectively. Limits of detection were 0.0382 and 0.14811 μg/ ml and limits of quantification were 0.1159 and 0.4488 μg/ml for ascorbic and gallic acid, respectively. Recovery was 99.60 - 100.28% and 100.26 -101.35% for ascorbic and gallic acid, respectively. The stress degradation studies were performed for both API and Ayurvedic as per ICH guidelines, the degradation was observed in oxidative, photolytic, hydrolytic degradation under acidic, alkaline conditions and dry heat induced studies. The proposed method is rapid, simple and also it can be applied for the routine analysis of herbal formulations.