Abstract

DEVELOPMENT AND VALIDATION OF AN RP-HPLC METHOD FOR THE ESTIMATION OF BOERAVINONE B IN BOERHAAVIA DIFFUSA EXTRACT

Ukande A. D., Shinde R. N.*, Dr. Ingole R. D., Ture P. R., Shinde O. N.

ABSTRACT

Boerhaavia diffusa Linn. (Punarnava) is an important medicinal plant widely used in traditional ayurvedic medicine for the treatment of hepatic, renal, inflammatory, and urinary disorders. The plant contains several biologically active phytoconstituents, among which Boeravinone-B is considered a major marker compound responsible for its pharmacological activities. Standardization and quantitative estimation of such marker compounds are essential to ensure the quality, safety, and therapeutic efficacy of herbal formulations. The present study was aimed at the development and validation of a simple, precise, accurate, and reproducible Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method for the quantification of Boeravinone-B in the hydroalcoholic extract of Boerhaavia diffusa. The hydro alcoholic extract of Boerhaavia diffusa was prepared and subjected tophytochemical screening and chromatographic analysis. RP- HPLC analysis was performed using an Agilent Zorbax C18 column (250 × 4.6 mm, 5 μm) with gradient elution consisting of phosphate buffer and acetonitrile as mobile phase. The flow rate was maintained at 1.5 mL/min and detection was carried out at 270 nm using a PDA detector. Boeravinone-B showed a well-resolved sharp peak at a retention time of approximately 24.413 minutes. The developed method was validated according to ICH guidelines for parameters including linearity, specificity, precision, and system suitability. The calibration curve exhibited excellent linearity within the concentration range of 1.02–4.01 μg/mL with correlation coefficient (R²) close to 0.999. The percentage relative standard deviation (%RSD) values were found to be within acceptable limits, indicating good precision and reproducibility of the method.

Keywords: Boerhaavia diffusa, Boeravinone-B, RP-HPLC, Herbal Standardization, Method Validation, Phytochemical Analysis, HPTLC, Punarnava.