Standardization and Evaluation of Triphala Juice and Quantification of Gallic Acid as a Biomarker by Analytical Techniques

Abstract

Standardization of Triphala Juice was performed by using the WHO Guidelines. The Parameters included Preliminary Analysis, Phytochemical Identification, Heavy Metal Estimation, etc. A new simple, specific, precise and accurate UV Spectrophotometric, High-Performance Liquid Methods: Chromatography and High-Performance Thin Layer Chromatography method has been developed for the Estimation of Gallic Acid in pure form. The UV- Spectrophotometric method was developed using Schimadzu 1800 UV - Visible spec-trophotometer using methanol as a solvent. The method was shown to be linear, with a detection wavelength of 273 nm for Gallic Acid. The separation was achieved on the Schimadzu Prominence-I RP-HPLC and the column used was C18 column using mobile phase consisting of mixture of Methanol: 0.1% OPA (50:50). The detection was carried out at 280 nm with a flow rate of 0.7ml/min. The retention time for Gallic Acid was found 3.89 minutes. The calibration curve was found linear (r2 = 0.999) for RP- HPLC method. The HPTLC method was developed using Aetron Sprayline instrument, Methanol as solvent and mobile phase consisting of Toluene: Ethyl Acetate: Formic Acid: Methanol (3:3:0.9:0.2). The method was found linear and the wavelength of detection for Gallic Acid was 254nm, respectively. The percentage recoveries for both methods were found in the 98.0- 102.0% range. The methods were validated in accordance with International Conference on harmoniza-tion acceptance criteria for specificity, linearity, precision, accuracy, robustness and system suita-bility. The excipients did not interfere in the determination of Gallic acid in Triphala Juice. The suggested approach was effectively implemented for the quantitative determina-tion of gallic acid in Triphala juice, which would aid in quality control