PHARMACEUTICO-ANALYTICAL STUDY OF RASAMANIKYA AND ITS IN VITRO CYTOTOXIC ACTIVITY ON A431 SKIN CANCER CELL LINE

Introduction: Skin cancer is among the most common malignancies worldwide, and squamous cell carcinoma constitutes a major type of non-melanoma skin cancer. In Ayurveda, most dermatological disorders are described under the broad concept of Kushtha. Rasamanikya, a classical Ayurvedic herbo-mineral formulation prepared from Shodhita Hartala, is indicated in disorders such as Kushtha, Vicharchika, Nadi Vrana, and Dustha Vrana. Therefore, the present study was undertaken to evaluate the in vitro cytotoxic activity of Rasamanikya on A431 skin cancer cell line. Aim: To prepare and analyze Rasamanikya according to Rasendra Chintamani and to study its in vitro cytotoxic activity on A431 skin cancer cell line. Materials and Methods: Rasamanikya was prepared from Shodhita Hartala processed in Kushmanda Swarasa and Amla Dadhi as per the classical method mentioned in Rasendra Chintamani using Antardhoom procedure in closed Sharava. The prepared formulation was subjected to organoleptic, physicochemical, X-ray fluorescence (XRF), X-ray diffraction (XRD), and particle size analysis. In vitro cytotoxic activity was assessed on A431 human skin cancer cell line using MTT assay at concentrations of 1000, 500, 250, 125, and 62.5µg/mL. Results: Rasamanikya was obtained as a light yellowish-colored, odorless, tasteless fine powder. Physicochemical analysis showed moisture content 0.16%, total ash 3.16%, acid insoluble ash 1.96%, water soluble extractive 1.46%, alcohol soluble extractive 1.55%, pH 5.6, specific gravity 0.998, and volatile matter 96.67%. XRF analysis revealed arsenic (64.4%) and sulphur (28.3%) as major constituents. XRD analysis demonstrated transformation of raw Hartala from orpiment (As₂S₃) to dimorphite (As₄S₃) form in Rasamanikya. Particle size analysis showed a primary particle size around 385 nm with Z-average of 686 nm and polydispersity index of 0.602. The MTT assay demonstrated dose-dependent cytotoxic activity against A431 cell line with progressive reduction in cell viability at increasing concentrations. Rasamanikya exhibited an IC₅₀ value of 197.6 µg/mL. Conclusion: The present study establishes successful preparation and analytical standardization of Rasamanikya. The formulation exhibited significant dose-dependent cytotoxic activity against A431 skin cancer cell line, indicating its potential anticancer activity. Further in vivo studies and mechanistic investigations are required to validate its therapeutic applicability in skin cancer management.