Background: The quality, safety, and efficacy of herbal medicines rely heavily on proper identification and standardization. Even after accurate botanical identification, herbal drugs can be of substandard quality due to faulty collection practices, incorrect storage conditions, or improper preparation methods.

Objectives: The present study aimed to establish the physico-chemical standards, perform qualitative phytochemical screening, and develop a Thin Layer Chromatography (TLC) fingerprint for the heartwood of Padmaka (Prunus cerasoides D. Don.) to ensure its quality, purity, and authenticity.

Methods: The physico-chemical analysis of the Prunus cerasoides sample was conducted at the drug testing laboratory (ISM), Joginder Nagar. The sample was evaluated for foreign matter, loss on drying, total ash, acid-insoluble ash, water-soluble extractive (WSE), and alcohol-soluble extractive (ASE) values according to standard pharmacopoeial methods. Qualitative phytochemical screening was performed on the heartwood powder to detect the presence of primary and secondary metabolites, including alkaloids, flavonoids, steroids, and saponins. Furthermore, TLC was utilized as a qualitative separation technique to establish a characteristic chemical fingerprint using a Toluene:Ethylacetate (9:1) mobile phase.

Results: The sample was completely free from foreign matter. Physico-chemical evaluation revealed a total ash value of 0.54% and an acid-insoluble ash value of 0.38%, both well below the maximum limits set by the Ayurvedic Pharmacopoeia of India (API). The extractive values were notably high, with a WSE of 30.34% and an ASE of 15.79%.

Conclusion: The analyzed sample of Prunus cerasoides D. Don. meets all standard physico-chemical parameters laid out in the API, indicating its high purity and quality. The rich presence of therapeutic secondary metabolites (flavonoids, steroids, and alkaloids) strongly correlates with its traditional Ayurvedic indications for skin and bleeding disorders.