Integrated Pharmacognostic, Phytochemical, and HPTLC Investigation of Hibiscus hispidissimus Griff. Leaves and Stem with Emphasis on β-Sitosterol

Abstract BACKGROUND: Hibiscus hispidissimus Griff. is a traditionally valued medicinal plant widely used by tribal healers in Kerala, India, for the treatment of inflammatory disorders, helminthic infections, and liver-related ailments. Despite its ethnomedicinal significance, there is a lack of systematic pharmacognostic and phytochemical standardization. This study aimed to establish comprehensive botanical, pharmacognostic, physicochemical, and phytochemical profiles of the leaves and stem of H. hispidissimus, including quantification of total phenolic content (TPC), total flavonoid content (TFC), and high-performance thin-layer chromatography (HPTLC) fingerprinting with marker compound analysis. METHODS: Collected plant specimens were subjected to macroscopic and microscopic examinations to establish diagnostic pharmacognostic characteristics. Physicochemical parameters and preliminary phytochemical screening were performed for both leaves and stem. Quantitative estimation of TPC and TFC was carried out using spectrophotometric methods. HPTLC fingerprinting was conducted using a solvent system of benzene: ethyl acetate: formic acid (6:4:0.5 v/v/v), and β-sitosterol was detected and quantified as a marker compound. RESULTS: The study revealed distinct differences between the leaves and stem in terms of physicochemical properties and phytochemical composition. HPTLC fingerprinting demonstrated band variation at 254 nm (Leaves: 13 bands; Stem: 9 bands), 366 nm (Leaves: 14 bands; Stem: 12 bands), and post-derivatization at 545 nm (Leaves: 15 bands; Stem: 14 bands). Quantitative analysis showed that the leaves contained higher TPC (12.16 ± 0.57 mg/g) than the stem (6.47 ± 0.05 mg/g), the leaves also exhibited significantly higher TFC (6.43 ± 0.05 mg/g) compared to the stem (0.21 ± 0.01mg/g). β-Sitosterol was present in both plant parts, with a higher concentration in the stem (10.72 ± 0.3195 µg/100 mg) than in the leaves (8.76 ± 0.3553 µg/100 mg). CONCLUSION: This investigation highlights the pharmacognostic and phytochemical distinctions between the leaves and stem of H. hispidissimus , emphasizing the differential distribution of bioactive constituents. The leaves are richer in phenolic compounds, while the stem contains more flavonoids and β-sitosterol. These findings provide a scientific foundation for the standardization and potential therapeutic applications of H. hispidissimus in traditional and modern herbal medicine.