Development and Validation of Analytical Method for Standardisation of Curcumin Using RP-HPLC Method in Prepared Extract

Abstract

Objective: The main objective of present work is to develop and validate simple, precise and accurate analytical method for identification of curcumin using RP-HPLC method in prepared extract. Methods: Spectrophotometric determination was performed on a Perkin-Elmer UV-VIS Double Beam Spectrophotometer to know the maximum absorbance of the compounds. Chromatographic separation was achieved using merck C18 analytical column (5 µm, 250 mm x 4.6 mm, i.d). The optimized solvent system consisted of methanol, acetonitrile in the ratio of 30:70 v/v. The effluents were detected by means of UV detector at 415nm. The proposed method was validated in accordance with International Conference on Harmonisation (ICH) guidelines. Results: Linearity was observed at a concentration range of 2.5-15µg/ml with a regression equation of y = 77703x – 586.13 and correlation coefficient of 0.9993. The retention time was found to be at 1.72min by means of the proposed method. The method was validated according to ICH guidelines and was found to contain the %RSD values below 2% which shows that the method was precise, accurate and specific. Conclusion: The developed method was validated as per the ICH guidelines which show that the method is sensitive, simple, precise and accurate. Thus the method is applied for the identification of curcumin in prepared extracts.