ABSTRACT Hygrophila spinosa is a medicinal plant traditionally used for its therapeutic properties, attributed to various bioactive phytoconstituents, including lupeol and quercetin. Accurate quantification of these compounds is essential for standardizing plant‐based formulations and ensuring quality control. The objective of this study was to develop and validate a simple, rapid, and accurate high‐performance liquid chromatography method for quantifying lupeol and quercetin in the hydroalcoholic seed extract of H. spinosa . The chromatographic analysis was performed on a Shimadzu system equipped with a C18 column. The mobile phase consisted of methanol and 0.1% o‐phosphoric acid (70:30 v/v), with a flow rate of 1.0 mL/min. Detection was carried out at 210 nm for lupeol and 370 nm for quercetin. Retention times were compared between standard solutions and extract samples. The method was validated following the International Conference on Harmonisation guidelines for parameters such as linearity, precision, accuracy, specificity, limit of detection, and limit of quantification. Lupeol and quercetin were effectively separated, with retention times of 4.3 and 5.5 min, respectively. The concentrations of lupeol and quercetin in the hydroalcoholic seed extract were found to be 13.68 and 16.50 mg/100 g, respectively. Calibration curves were linear over the concentration range of 20–70 µg/mL for lupeol and 50–100 µg/mL for quercetin. The validated HPLC method demonstrated excellent precision, accuracy, and sensitivity for quantifying lupeol and quercetin in H. spinosa seed extract. This method is suitable for routine quality control and standardization of formulations containing this plant. Future studies may explore more advanced extraction procedures and in vivo evaluation of the therapeutic potential of the bioactive.
GAIKWAD et al. (Thu,) studied this question.