Validation, Optimization and Hepatoprotective Effects of Boeravinone B and Caffeic Acid Compounds from Boerhavia diffusa Linn

Boerhavia diffusa, also known as Punarnava, is a plant of the Nyctaginaceae family that has been utilized in traditional medicine to cure a variety of ailments. The goal of this study was to use response surface methodology (RSM) to optimize the maximum percentage yield of boeravinone B and caffeic acid from Boerhavia diffusa roots, and simultaneous determination of boeravinone B and caffeic acid in newly developed single solvent system and demonstrate the hepatoprotective benefits of boeravinone B and caffeic acid. The extraction process examined extraction time, extraction temperature and solvent concentration, which were optimized via Box–Behnken experimental design. The proposed HPTLC method for the quantification of boeravinone B and caffeic acid were successfully validated and developed. The method was validated in term of linearity and detection limit, quantification limit, range, precision, specificity and accuracy. The separation of boeravinone B and caffeic acid bands was achieved on HPTLC plate using formic acid: ethyl acetate: toluene (1:3:5 v/v) as developing system. Densitometric analyses of boeravinone B and caffeic acid was carried out in the absorbance mode at 254 nm. The maximum percentage yield of caffeic acid and boeravinone B from Boerhavia diffusa require appropriate extraction parameters such as temperature, time, organic solvents and water content, which can be achieved using the Box-Behnken statistical design provide time: temperature: solvent ratio (30:45:40 v/v) for extraction of caffeic acid and 60:60:40 v/v for extraction of boeravinone B. The boeravinone B (200 µg/mL) and caffeic acid (200 µg/mL) showed the most significant hepatoprotective activity compared with standard sylimarin in HepG2 cell induced with galactosamine 40 mM toxicity. The findings supported B. diffusa’s traditional use as a functional food forhuman health benefits.