Abstract
Background: Madhuca longifolia is a medicinally significant plant widely used in Ayurveda for the management of wounds, inflammation, pain, and dermatological disorders. With the growing adoption of topical herbal formulations, particularly spray-based delivery systems, rigorous phytochemical standardisation is essential to ensure therapeutic consistency, quality control, and reproducibility. Objective: The present study aimed to perform phytochemical characterisation and chromatographic standardisation of M. longifolia flower extract using High-Performance Thin-Layer Chromatography (HPTLC), with particular emphasis on constituents associated with anti-inflammatory and analgesic activities Methods: The ethanolic extract of M. longifolia flowers was subjected to HPTLC fingerprinting using silica gel 60 F₂₅₄ as the stationary phase. Chromatographic development was carried out in a CAMAG twin trough chamber using toluene: ethyl acetate: formic acid (6:3:0.5) as the mobile phase. Detection was performed under ultraviolet light at 254 nm and 366 nm, and Rf values were recorded. Phytochemical classes were tentatively identified based on chromatographic behaviour and correlation with published literature. Results: HPTLC fingerprinting revealed multiple well-resolved phytochemical bands at 254 nm, indicative of phenolic acids, flavonoids, terpenoids, sterols, coumarins, and related constituents, while UV detection at 366 nm confirmed the presence of flavonoid and phenolic compounds. These phytochemical classes are widely reported for their analgesic, anti-inflammatory, and wound-healing activities. Conclusion: The established HPTLC fingerprint offers a reproducible analytical profile for M. longifolia flower extract, substantiating its traditional therapeutic applications and providing a scientific basis for the standardisation of topical Ayurveda formulations.

Key words: Madhuca longifolia flower extract, Mahua , HPTLC, Phytochemistry