DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF RESVERATROL AND CURCUMIN IN HERBAL FORMULATION
*Shaheen Sheikh and Vandana Jain
ABSTRACT
A simple, precise, cost effective RP-HPLC method has been developed and validated for the determination of Resveratrol and Curcumin in Herbal formulation. The chromatographic separation was achieved using a Prontosil C18 column (250 x 4.6mm, 5μm) and mobile phase consisting of acetonitrile: 0.02M potassium dihydrogen phosphate buffer (60:40), pH adjusted to 3.0 with Orthophosphoric acid at flow rate 1ml/min was used. Detection wavelength was set at 320 nm for both the drugs. The retention time of Resveratrol and Curcumin were found to be 2.703 and 6.553 minute. The resolution of Resveratrol and Curcumin peak was found to be more than 2. The calibration curves showed good linear correlation coefficients (r2 > 0.9997) within the tested ranges. The recoveries were found to be between 99.2-101 % for Resveratrol and 98.9-100.8 % for Curcumin. The %RSD for precision and accuracy of the method was found to be less than 2%. The developed method was successfully used for the quantitative estimation of these two markers in a marketed herbal formulation.
Keywords: Resveratrol, Curcumin, simultaneous determination, RP-HPLC, validation, ICH guidelines.
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