Validation of an optimized HPLC–UV method for the quantification of formononetin and biochanin A in Trifolium pratense extract

Abstract Here, two important isoflavones present in Trifolium pratense, formononetin and biochanin A, were analyzed by high performance liquid chromatography coupled with UV (HPLC–UV). These are marker compounds anti-inflammatory, anti-cancer, anti-oxidant properties, also potent phytoestrogens that can be used hormonal therapies. Till date, no study regarding rapid identification quantification of T. pratense extract has been conducted. This developed validated an optimized method for quantifying A using HPLC–UV. The results showed excellent linearity the calibration curve (r 2 ≥ 0.999), good resolutions chromatographic peaks obtained. Other validation parameters such as specificity, accuracy, precision demonstrated our had reliability sensitivity. Furthermore, is convenient.