Development and validation of Stability Indicating HPLC method for determination of Ellagic and Gallic acid in Jambul seed
Keywords:Ellagic acid (EA), Gallic acid (GA), High Performance liquid Chromatography, Stability indicating method.
Ellagic and Gallic acid are main phytoconstituents of S.cumini seeds. These are the phenolic compounds. An approach for the stress degradation was successfully applied for the development of stability indicating HPLC method for the determination of Ellagic and Gallic acid. Sample was resolved on a Hypersil C18 (250*4.6 mm particle size 5?) column. The mobile phase consisted of 1% OPA and ACN and in the ratio of 70:30 v/v which was sonicated to degas and delivered at a flow rate of 1ml/min at ambient temperature. The retention time of Ellagic acid and Gallic acid was 3.1±0.05 & 4.1±0.05 minutes. Studies were performed using an HPLC system equipped with a UV detector; the response was monitored at 271nm. The method is specific to Ellagic and Gallic acid; it is able to resolve the peak from ethanolic extract of s.cumini seeds and formulation. The calibration curve was linear over the concentration range of 8-24 ?g/ml (r2=0.997, 0.998 resp). The limit of detection for Ellagic acid and Gallic acid was found to be 0.25?g/ml, 0.15?g/ml resp. and the quantification limit was about 0.75?g/m, 0.49?g/ml. The accuracy of the method was established based on the recovery studies. The markers were subjected to acid, base, neutral hydrolysis, oxidation, thermal degradation and photolysis. The method was successfully validated according to ICH guidelines Q2 (R1).
Int J Appl Sci Biotechnol, Vol 3(3): 434-438