Standardization and High-performance Thin-layer Chromatography Fingerprinting of Unani Formulation Habb-e-Zahar Mohra

Background: Habb-e-Zahar Mohra is a classical herbo-mineral Unani formulation traditionally used for its qābiḍ (astringent) action in managing conditions such as Ishal-e-Atfal (diarrhea in children) and Atash-e-Mufrit (excessive thirst). Despite its therapeutic applications, the lack of standardization and quality assessment poses challenges to its efficacy and safety. Objective: This study aimed to standardize and evaluate the physicochemical, phytochemical, and chromatographic characteristics of Habb-e-Zahar Mohra to ensure its quality, safety, and reproducibility in adherence to Unani pharmacopoeial standards. Materials and Methods: The formulation was prepared following the National Formulary of Unani Medicine guidelines. Organoleptic, physicochemical (pH, moisture content, ash values, and extractive values), and phytochemical properties were evaluated. High-performance thin-layer chromatography (HPTLC) fingerprinting was performed using extracts in petroleum ether, chloroform, methanol, and aqueous solvents to establish a fingerprint profile for quality control. Results: Organoleptic and physicochemical properties: The prepared formulation exhibited consistent organoleptic characteristics (color: clay, odor: agreeable, texture: uniform) and physicochemical properties. The pH was 6.30, moisture content was 8.1%, total ash value was 8.31%, acid-insoluble ash was 6.15%, and water-soluble ash was 0.12%. Extractive Values: Extractive values were highest in aqueous extract (12.65%), followed by methanol (4.26%), chloroform (4.02%), and petroleum ether (1.85%). HPTLC fingerprinting: HPTLC analysis revealed distinct peaks under ultraviolet light at 254 nm and 366 nm, with variations in Rf values and area percentages across solvent extracts. Methanol and aqueous extracts showed the highest number of peaks, indicating a rich phytochemical profile. Conclusion: The study successfully standardized Habb-e-Zahar Mohra , ensuring its consistency, quality, and safety. HPTLC fingerprinting established a reliable chemical profile, providing a scientific basis for the formulation’s therapeutic claims. This approach supports the global acceptance of traditional Unani medicine by aligning with modern quality control practices.