Exploring Bioactive Phytochemicals of Hygrophila difformis Blume: Extraction, Isolation, and GC–MS Characterization

Hygrophila differformis Blume is a traditional plant of scientific interest due to the reported wide range of secondary metabolites. Systematic phytochemical profiling of its solvent fractions is essential to map phytoconstituent distribution, identify bioactive markers, and guide focused pharmacological investigations. Sequential solvent extraction with hexane and ethyl acetate was performed, and the fractions were separated with silica‐gel column chromatography to ascertain numerous fractions. GC–MS of the hexane fractions showed that long‐chain aliphatic hydrocarbons and primary alcohols were the major phytochemicals. Squalene and phytol were terpenoid compounds in significant amounts, which could have some biological importance. Both saturated and unsaturated hydrocarbons and primary alcohols were enriched in the ethyl acetate fractions, and the formation of compounds like neophytadiene and cis, cis, cis‐7,10,13‐hexadecatrienal was only observed in these fractions. Both hexane and ethyl acetate extracts contained fatty acids (n‐hexadecanoic acid), but only ethyl acetate fractions contained linoleic acid and α ‐linolenic acid. A further indication of the pharmacological potential of the plant is the identification of stigmasterol (4.76%) as a marker phytosterol. The GC–MS profiling of hexane and ethyl acetate extracts revealed several bioactive phytochemical classes, which may serve as a chemical reference for future studies focusing on compound isolation, structural characterization, and biological evaluation of this relatively underexplored species.