A Validated HPTLC Method for Quantification of Linoleic Acid and Beta-Sitosterol in Solanum nigrum Extract

Solanum nigrum, a member of the Solanaceae family, has long been used as both food and medicine. This plant has been recognized for its immunomodulatory, anti-ulcerogenic, hepatoprotective, and anti-hyperlipidemic effects. It contains various chemical components such as alkaloids, coumarins, flavonoids, tannins, saponins, proteins, carbohydrates, glycosides, and phytosterols. The objective of the current work was to create a repeatable and consistent HPTLC-validated technique for concurrently detecting Beta-Sitosterol (BTS) and Linoleic Acid (LIA) in S. nigrum in accordance with ICH recommendations. The HPTLC process involved using silica gel 60 F254 TLC plates as the stationary phase and a mobile phase consisting of n-hexane, ethyl acetate, and formic acid in a ratio of 6:4:0.2 (v/v/v). The resulting bands were uniformly visualized after derivatization with anisaldehyde sulfuric acid. The calibration curves of standard LIA and BTS exhibited satisfactory linearity within concentration ranges of 400-1150 ng/spot and 350-1200 ng/spot, respectively, with correlation coefficients (r2) of 0.99153 and 0.99389. LOD and LOQ were determined as 91.96 and 118.14 ng/spot for LIA, and 278.66 and 314.62 ng/spot for BTS, respectively. In conclusion, this HPTLC method proved to be efficient, simple, precise, and reproducible for measuring LIA and BTS in S. nigrum.