AYUSH‐64, a Coded Polyherbal Formulation: High‑Performance Thin‑Layer Chromatographic Standardization, Quantification of Marker Compounds, GC–MS Profiling, and In Silico Studies

Standardization of any herbal drug is crucial for maintaining quality control, enhancing product safety, and ensuring that the drug or formulation performs as intended. In the present study, AYUSH‐64 was standardized using high‐performance thin‐layer chromatography (HPTLC) in detail with respect to its raw drugs and their aqueous extracts, and quantified two marker compounds, ursolic acid and lupeol. Additionally, GC–MS analysis of the ethyl acetate and acetonitrile extracts confirmed the presence of 13‐docosenamide, (Z)‐γ‐sitosterol, β‐amyrin, lup‐20(29)‐en‐3‐ol acetate, and (3β)‐components in both extracts using the NIST library. Furthermore, in silico studies were conducted on lupeol and ursolic acid to predict their interactions with the ACE2 membrane protein. The study demonstrated favorable binding interactions between the identified phytochemicals and the ACE2‐targeting monoclonal antibody (7V61), indicating their potential as broad‐spectrum coronavirus‐blocking candidates. The observed binding energies of −6.58 kcal/mol and −6.21 kcal/mol suggest appreciable binding affinity and stable interactions within the predicted binding site. These findings provide encouraging preliminary evidence supporting the potential biological relevance of these phytochemicals and establish a strong basis for further investigation. Future studies involving in vitro, cell‐based experiments, and other biological validation approaches will be valuable for confirming these computational predictions and further elucidating their therapeutic potential.