Validation, Optimization and Hepatoprotective Effects of Boeravinone B and Caffeic Acid Compounds from Boerhavia diffusa Linn
Kamal Y. Thajudeen, Abdulrhman Alsayari, Shehla Nasar Mir Najib Ullah, Shahana Salam, Muhammed Elayadeth‐Meethal, Ilyas Uoorakkottil
Abstract
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.