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Abstract
Stevia rebaudiana Bertoni (Asteraceae) is a perennial shrub of South America, produces diterpene glycosides used as low calorie sweeteners. Stevioside and extracts prepared from the leaves have been used widely all over the world as sweetening agents, taste modifiers and sugar substitutes. Gallic acid (GA) is reported for its anti-inflammatory, anticancer, antiviral and antimelanogenic activities and many of these are mediated by its antioxidative capacity. Presently a study has been conducted on the comparative High Performance Thin Layer Chromatography (HPTLC) analysis of gallic acid from the dried leaf powder of in-situ and in-vitro grown S. rebaudiana. Nodal explants of S. rebaudiana were cultured in vitro in MS media supplemented with 6-benzyladenine (BA; 8.5 mM) and indole-3-butyric acid (IBA; 9.2 mM). The quantitative determination of gallic acid was performed on silica gel 60 F254 HPTLC plates as stationary phase. The linear ascending development was carried out in a twin trough glass chamber saturated with a mobile phase consisting of chloroform: ethyl acetate: formic acid (5:4:1) at room temperature (22±2°C). CAMAG TLC scanner-3 equipped with CATS software (version: 1.4.4.6337) was used for spectrodensitometric scanning and analysis in the ultraviolet region at λ=280 nm. The method was validated for linearity, precision and accuracy. Correlation coefficient, limit of detection (LOD), limit of quantification (LOQ) as well as recovery values were found to be satisfactory. S. rebaudiana raised in vitro (0.042 %) was found to contain higher amount of GA than their naturally growing counterparts (0.022 %).