Phytosterols are bioactive compounds that are naturally present in plant cell membranes with a similar chemical structure to the mammalian cell- derived cholesterol. β-sitosterol stands out as the primary phytosterol compound abundant in plants. The reason it is present in so many plant and vegetable oils is due to its steroidal lipophilic nature. From various in-vitro and in vivo studies it is found to possess important activities as Anticancer, Antioxidant, Antimicrobial, immunomodulatory, Anti-inflammatory, lipid lowering, hepato-protection. A properly developed and validated method of quantification can be used for phytochemical profiling and estimation of the standard compound. Analysts are able to improve their comprehension and tailor their own experimental strategies for drug analysis by thoroughly evaluating the procedures used in earlier research. Analysts who are looking for information on the precise set of instruments and solvents used in the analysis of beta sitosterol may find this review useful. Thus article aims to exhaustively explore analytical methods for β-sitosterol, encompassing methods such as GC-MS, HPLC, and HPTLC.