SECONDARY METABOLITE SCREENING OF Marsilea minuta L. FOR ALKALOID CONTENT: VALIDATION THROUGH TRIPLE CHROMOGENIC PRECIPITATION TECHNIQUES

Marsilea minuta L., commonly known as dwarf water clover and belonging to the Marsileaceae family, has established significance in traditional medicinal practices across various cultural systems, yet comprehensive phytochemical characterization of its alkaloid constituents remains substantially underexplored in contemporary scientific literature. This investigation aimed to systematically evaluate and assess alkaloid compounds present in methanolic extracts of Marsilea minuta through controlled extraction protocols and classical qualitative screening methodologies. Authenticated plant specimens underwent methanolic maceration under precisely controlled temperature conditions ranging from 25-30°C for an extended 72-hour period, followed by sequential filtration processes and systematic concentration procedures utilizing digital water bath evaporation at 80°C for three hours, culminating in final concentration at 67.5°C for one hour to achieve optimal extract consistency. The presence of alkaloid compounds was definitively confirmed through the application of three established classical precipitation tests, specifically Mayer's reagent test, Wagner's reagent test, and Dragendorff's reagent test, each targeting different alkaloid structural characteristics. Comprehensive screening results demonstrated consistently positive reactions across all three testing methodologies, with Mayer's test producing distinctive cream-colored precipitates, Wagner's test yielding characteristic reddish-brown precipitates, and Dragendorff's test exhibiting pronounced orange-red precipitates, collectively providing robust evidence for the presence of diverse alkaloid compounds within the methanolic extract. The successful methanolic extraction protocol effectively yielded alkaloid-rich extracts from Marsilea minuta plant material, as conclusively evidenced by positive reactions to all standard alkaloid detection reagents employed in this study. These preliminary findings establish crucial foundational data for subsequent detailed alkaloid characterization studies, structural elucidation investigations, and potential pharmaceutical applications, while supporting the ethnopharmacological significance attributed to this aquatic fern species. The confirmed alkaloid presence validates the therapeutic potential of Marsilea minuta and opens avenues for advanced analytical characterization and bioactivity evaluation studies.