Development and Validation of a RP-HPLC Method for the Determination of Ellagic Acid in Terminalia bellirica Extract and Single Herb Capsule of Terminalia bellirica

Abstract:

Objective: The present work describes development and validation of a simple, specific, sensitive, precise, reproducible and robust high performance liquid chromatographic method of analysis of Ellagic acid, as in Terminalia bellirica extract and from capsule formulation. A validated rapid HPLC–PDA method was developed for identification and quantification of Ellagic acid (EA) in the extracts prepared from the fruits of Terminalia bellirica available in India. Methodology: The separation was achieved on a Waters HPLC system coupled with a PDA detector. The separation was achieved by using mobile phase in composition of n-Hexane sulfonic acid (20 mM) and 100% Methanol (Gradient programme) at flow rate of 0.7 ml/min was used. Results: The retention time for Ellagic acid was found to be 21 min. The system suitability parameters were calculated and were found to be within limits. Linear relationship was obtained between response and amount of drug with high correlation coefficient (R2) in the range 250-750 ppm for Ellagic acid (R2=0.9998).The results of formulation analysis were validated as per International Council of Harmonization (ICH) guidelines indicating high degree of accuracy. Conclusion: The % R.S.D. value which is less than to 2 for robustness study suggests that the developed HPLC methods are unaffected by small changes in process parameters.

Key words: Terminalia bellirica, Ellagic acid, RP-HPLC, Validation, ICH Guidelines.