Future-proofing RP-HPLC method for quantification of beta-sitosterol in Pygeum extract and cholecalciferol: Promising approaches for prostate health and dietary supplementation

Abstract The preponderance of benign prostatic hyperplasia (BPH) and prostate cancer has increased in prevalence among aging men; prostate health management is becoming increasingly important. Studies indicate that substances such as beta-sitosterol and cholecalciferol may have therapeutic benefits for prostate disorders. This study aims to innovate the HPLC approach for the coexisting quantification of beta-sitosterol in Pygeum extract ( Prunus africana ) and cholecalciferol, providing a comprehensive strategy for evaluating the quality of herbal products targeting prostate health. The RP-HPLC technique was appraised using modified green star area (MoGSA) and BAGI metrics based on an Inertsil ODS-3 C18 column with a mobile phase of methanol and acetonitrile in a 95:5 ratio and detection at 210 nm. The strategy exhibited outstanding resolution, precision, and robustness, with a 12-min runtime. The method's linearity was confirmed with R 2 above 0.99, and the LOD and LOQ values were 0.61 μg mL −1 , 0.76 μg mL −1 , and 1.85 μg mL −1 , 2.31 μg mL −1 for beta-sitosterol and cholecalciferol, respectively. Precision intra- and inter-day testing revealed RSD below 2%, confirming the method's reliability. Recovery studies indicated high accuracy, with recoveries of 101.50, 100.99, and 101.17, 100.92% in the pharmaceutical formulations and the supplement formulations for beta-sitosterol and cholecalciferol, respectively. This innovative dual-action approach may provide a promising therapeutic strategy by targeting multiple pathways involved in prostate function and disease prevention.