Identification of Anticholinesterase Active Compounds from the Ethylacetate Fraction of Hydroalcoholic Extract of Itrifal Sana Using TLC–bioautography–MS and Its Validation Using an In Silico Molecular Approach.

This article focuses on the phytochemical characterization and acetylcholinesterase (AChE) inhibitory potential of Itrifal Sana (IS), a traditional Unani polyherbal formulation used for various health conditions. Using a combination of thin-layer chromatography (TLC)–bioautography–mass spectrometry (MS), ultra-performance liquid chromatography-mass spectrometry (UPLC–MS), and in silico molecular docking, the study identified rosmarinic acid, apigenin, and catechin as key bioactive compounds responsible for AChE inhibition, with lobeline and rosmarinic acid showing the highest binding affinity in computational models. These findings suggest that IS may have therapeutic potential in managing Alzheimer's disease by inhibiting AChE, although further in vitro, in vivo, pharmacokinetic, and toxicity studies are needed to fully validate its efficacy and safety. The study also highlights the utility of integrated chromatographic and computational approaches for quality control and bioactivity assessment of traditional herbal medicines.