A comprehensive standardization of a polyherbal Unani formulation: phytochemical screening, HPTLC fingerprint, an in vitro antioxidant and cell proliferation B16F10 cell line assay on hypopigmentation disorder

The global demand for traditional medicine, in particular Unani System of Medicine, requires the standardization of herbal drugs based on contemporary analytical parameters to ensure the quality, safety, and authenticity of classical Unani formulations. The study aimed to standardize the Ḥabb-e-Hindi (HH) and to meet the requisite standards in the polyherbal formulations for acceptability, safety, high quality, and efficacy as per WHO to prove scientifically. The HH was standardized and evaluated through pharmacognostic, phytochemical screening, and physicochemical parameters. HPTLC fingerprint profile helps in quality control developed in ethanolic extract showed bands at specific R f values indicates the presence of bioactive compounds. To evaluate the antioxidant scavenging assay such as ABTS, FRAP, DPPH, using validated methodologies. To evaluate cell cytotoxicity, melanin activity, tyrosinase activity of HH on B16F10 cell line was studied, and its effect on hypopigmentation disorder in vitro as no reports were available till date. The validated study for the antioxidant assay carried out using ascorbic acid (Merck, India) as standard are expressed in AAE/mL equivalents of extract. ABTS radical scavenging activity was similar in aqueous and ethanolic extract. In the FRAP method highest antioxidant capacity was shown by the methanolic extract (IC 50 107.77 μg/mL) whereas ethanolic extract (IC 50 185.18 μg/mL) in the DPPH method. Upon cell proliferation assay HH showed no cytotoxicity activity when applied on B16F10 cell lines in different solvent extracts showed IC 50 value 300–450 μg/mL. Further research is needed to identify the bioactive compounds and biological activities that are accountable for the therapeutic properties.