Development of an HPLC Analytical Method for β-Sitosterol Used as a Treatment of Burns and Skin Ulcers

This study focuses on developing a highly accurate and reproducible High-Performance Liquid Chromatography technique of β-sitosterol for the quantification. Widely, a compound used as a treatment of burns and skin ulcers. This developed method ensures reliable separation, identification, and quantification of β-sitosterol in pharmaceutical formulations. Accuracy was assessed through recovery studies at three levels (80%, 100%, and 120%). The acceptance criterion was a recovery rate between 98% and 102%, which was successfully met. The detection limit (LOD) is defined as the lowest concentration of an analyte that can be detected, though not necessarily quantified. Typically, a signal-to-noise (S/N) ratio of 3: 1 is considered acceptable for estimating the LOD. The quantification limit (LOQ) refers to the lowest concentration that can be quantitatively measured with acceptable precision and accuracy, usually determined at a signal-to-noise ratio of 10: 1. The LOD is calculated using the formula 3.3 × (Sy/x / b), while the LOQ is calculated as 10 × (Sy/x / b). In this method, the LOD was found to be 2 µg/mL, and the LOQ was 6 µg/mL. The LOD and LOQ were determined based on the standard deviation of the response (σ) and the slope (S) of the calibration curve. The calculated LOD was 1.52 µg/mL.