Simultaneous Quantitative HPLC Analysis of Ascorbic Acid, Gallic Acid, and Catechin in Punica granatum, Tamarindus indica and Prunus domestica
Analysis of antioxidant compounds in Punica granatum, Tamarindus indica and Prunus domestica is very important because of commercial interest in the plant material as herbal drugs and in extracts for the nutraceutical and pharmacological value . A reversed-phase high-performance liquid chromatographic method has been developed and validated for simultaneous estimation of ascorbic acid, gallic acid and catechin in Punica granatum L (Lythraceae), Tamarindus indica L (Leguminosae) and Prunus domestica L (Rosaceae). A C18 column was used with isocratic elution of 0.1% glacial acetic acid in HPLC-grade water and acetonitrile (70:30; v/v) as the mobile phase at a flow rate of 1.0 mL/min with 15 minute run time. UV detection was performed at 254nm. Validation of the method was performed in order to demonstrate its selectivity, linearity, precision, accuracy, limits of detection and quantification (LOD and LOQ) . Calibration plots were linear over the concentration ranges 1 – 500 µg/mL for ascorbic acid, 5 – 500 µg/mL for gallic acid, and 50 – 500 µg/mL for catechin with correlation coefficients 0.995, 0.995 and 0.997, respectively. Repeatability, intra-day, inter-day and inter-analyst 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 RP-HPLC method was simple, precise and accurate and can be used for the quality control of the raw materials as well as formulations. Acknowledgements: The financial support from the CCRUM, Department of AYUSH, Ministry of health and family welfare, Government of India, New Delhi India is duly acknowledged. References:  Azzi A, Davies KJA, et al (2004) FEBS letters 558: 3 – 6.  International conference on harmonization (1996) Validation of Analytical Procedures Methodology Q2B.