Development and Validation of RP-HPLC Method for Estimation of Epigallocatechin -3-gallate (EGCG) in Lipid based Nanoformulations

ABSTRACT:Epigallocatechin-3-gallate (EGCG) is a potent antioxidant and free radical scavenger present in green tea. It is extensively used as antioxidant, immunomodulator, anticancer, neuroprotective, cardioprotective, antiviral,antibacterial, photo protective and anti aging agent. The objective of the present work was to develop and validate an RP-HPLC method for determination of EGCG in lipid based nanoformulations. RP-HPLC analysis was performed by isocratic elution using a Phenomenex C18 column (4.6 × 250 mm, 5 µm) at 25 °C. A mixture of acetonitrile and 1% v/v acetic acid (25:75) of pH 2.8 was used as a mobile phase with a flow rate of 0.8 mL/min. The analysis was performed using a UV detector at 280 nm. The results showed that the peak area response was linear within the concentration range of 0.5-40 µg/mL (r =0.9998). The values of LOD (limit of detection) and LOQ (limit of quantification) were 0.125 µg/mL and 0.272 µg/mL respectively. The % RSD for peak area of intraday and interday precision was 0.08 and 0.2%, respectively. The results of accuracy for the different concentrations of EGCG (8, 10 and 12 μg/mL) were 100.29±0.19, 101.10±0.10 and 100.44±0.25 % respectively. The validated method was successfully applied for the estimation of EGCG content in lipid based nanoformulations.