Development and Validation of Novel Rp-HPLC Method for the Simultaneous Estimation of Ellagic Acid and Quercetin in an Ayurvedic Formulation

Abstract

Objective: To develop a novel, accurate, precise and linear reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous qualitative and quantitative estimation of ellagic acid and quercetin in an ayurvedic formulation and validate as per international conference on harmonization (ICH) guidelines.Methods: In the present work, good chromatographic separation was achieved isocratically using a shim-pack HPLC C18 column (4.6 x 250 mm, 5μm) and a mobile phase consisting of 0.02 M potassium dihydrogen orthophosphate buffer (pH adjusted to 3.5 with orthophosphoric acid) and acetonitrile in the ratio 60:40, at flow rate of 1.2 ml/min and column temperature maintained at 35 °C. The effluents obtained were monitored at 255 nm with UV-visible detector.Results: The retention time of ellagic acid and quercetin were found to be 1.65 min and 2.94 min respectively. Linearity of ellagic acid and quercetin were tested in the range of 6-14 ppm and 3-11 ppm respectively. The correlation coefficient for ellagic acid and quercetin were 0.997 and 0.993 respectively. The high recovery values (98 %-102 %) indicate a satisfactory accuracy. The low percent relative standard deviation (% RSD) values in the precision study reveals that the method is precise.Conclusion: The developed method is novel, simple, precise, rapid, accurate and reproducible for simultaneous quantitative estimation of ellagic acid and quercetin in an ayurvedic formulation. Hence the developed method can be used for quantitative analysis and quality control of extracts and commercial samples of other plant species and formulation containing these two markers.

Development and Validation of HPLC And HPTLC for Simultaneous Analysis of E and Z Guggulsterone, A-11–KBA And 11–KBA from Herbal Formulation

“…The continuous flow rate was 150 µL/s. 5 At room temperature, the linear growth was nurtured in a 20x10 cm glass twin trough container (25°C ± 2) and relative humidity of 60 ± 5% using a solvent system (17 mL) comprised of n-nexane, chloroform, ethyl acetate, and methanol (10:3:3:1, v/v/v). About 80 mm of the chromatogram was extrapolated from the position of the sample.…”

A Novel Rp-HPLC Method for Simultaneous Estimation of Berberine, Quercetin, and Piperine in an Ayurvedic Formulation

“…The literature survey reveals that spectrophotometric [6][7], highperformance thin layer chromatography (HPTLC) [8], reverse-phase high-performance liquid chromatography (RP-HPLC) [9][10] based method based methods have been reported for analysis of these drugs alone and in combination with other drugs. However, no HPLC method has been reported for simultaneous quantitative estimation of berberine, quercetin, and piperine in any ayurvedic formulation.…”