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Abstract

SIMULTANEOUS QUANTITATIVE HPLC ANALYSIS OF ASCORBIC ACID, GALLIC ACID, AND CATECHIN IN PUNICA GRANATUM, TAMARINDUS INDICA AND PRUNUS DOMESTICA USING BOXBEHNKEN STATISTICAL DESIGN

Mohd Amir, Mohd Mujeeb*, Sayeed Ahmad, Mohd Akhtar, Kamal YT, Kamran Ashraf

ABSTRACT

A reversed-phase HPLC method has been developed and validated for simultaneous estimation of ascorbic acid, gallic acid and catechin in P. granatum, T. indica and Prunus domestica. A C18 column was used with isocratic elution of 0.1% glacial acetic acid: acetonitrile (70:30; v/v) as mobile phase at a flow rate of 1.0 mL/min with 15 minute run time. Validation of the method was performed in order to demonstrate its selectivity, linearity, precision, robustness, accuracy, limits of detection and quantification. Calibration plots were linear over the concentration ranges 1-500μg/mL for ascorbic acid, 5-500 μg/mL for gallic acid, and 50-500 μg/mL for catechin with correlation coefficients 0.9953, 0.9951 and 0.9976, respectively. Precision RSDs of peak areas were less than 2.41%. The recovery of ascorbic acid, gallic acid and catechin in sample was found in the range from 98.88 to 102.50%. Limits of detection were 0.31, 1.53, 15.16 μg/mL and limits of quantification were 1.03, 5.12, 50.31 μg/mL for ascorbic acid, gallic acid and catechin, respectively. The boxbehnken statistical design was used for the validation of robustness. It showed that proposed method was simple, precise and accurate and can be used for the quality control of the raw materials as well as formulations.

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