Bala (Sida spp.), a key herb in Ayurvedic system of medicine, is traditionally used for its anti-inflammatory and analgesic properties, particularly in the management of arthritis and related musculoskeletal disorders. High-Performance Thin-Layer Chromatography (HPTLC) is an effective analytical technique for the quantitative evaluation of bioactive compounds. The present study focuses on the validation of an HPTLC method for the quantification of Caffeic acid and Vasicinone in the methanolic extracts of the roots of Sida cordifolia , Sida rhombifolia subsp. alnifolia, Sida acuta and Sida rhombifolia . Optimized mobile phases toluene: ethyl acetate: methanol: formic acid (6:4:1:0.3, v /v/ v /v) for Caffeic acid, and toluene: ethyl acetate: methanol (7:3:0.5, v/v/v) for Vasicinone provided good separation of the target compounds. The method was validated in accordance with ICH 2023 guidelines. The developed method exhibited excellent linearity, with correlation coefficients of 0.9996 for Caffeic acid (50–300 ng/band) and 0.997 for Vasicinone (100–500 ng/band). LOD for caffeic acid and vasicinone were found to be 15.27 and 33.2 ng/band respectively, while LOQ was noted as 46.28 and 100.7 ng/band. Recovery studies confirmed high accuracy, with percentage recoveries ranging from 98.6% to 99.9% for Caffeic acid and 99.4% to 99.8% for Vasicinone. • Bala (Sida spp.), a key herb in Ayurvedic system of medicine, is traditionally used for its anti-inflammatory and analgesic properties. • Densitometry based HPTLC method has been developed and validated in accordance with ICH guidelines for the quantitation of two active marker compounds such as Caffeic acid and Vasicinone in selected species used as source plant for Bala. • The developed method can be efficiently used for the quality standardization and to ensure the presence of therapeutically active compounds in selected species.
Saji et al. (Sun,) studied this question.