Aquatic plants, vital components of aquatic ecosystems, play pivotal roles in nutrient cycling, habitat provision, and overall ecological stability. This study presents a comprehensive investigation into the leaf biochemical composition and phytoconstituents of six distinct aquatic plant species viz. Nymphaea mexicana, Trapa natans, Nymphoides peltata, Azolla cristata, Salvinia natans and Ceratophyllum demersum, using standardized methodologies. The findings unveiled intriguing variations in the biochemical composition across the six aquatic plant species, highlighting their unique metabolic signatures. Soluble carbohydrate, carbon and total chlorophyll were found maximum in N. mexicana. Soluble protein, lipid, nitrogen, and ash content were found maximum in T. natans, N. peltata, A. cristata, and C. demersum, respectively, while moisture content was found maximum in S. natans. The difference in chemical composition in these species can be attributed to several factors, including their adaptation to aquatic environments and local variations in environmental conditions like light, temperature, nutrient and carbon dioxide availability to these plants. The phytoconstituent analysis through GC-MS uncovered a spectrum of bioactive compounds, including terpenoids, alkaloids, phenolic compounds, and fatty acids, in varying abundance across the six species, each having a number of pharmacological potentialities with numerous health and therapeutic benefits. This research expands our understanding of the biochemical diversity within aquatic plant species and underscores their potential as valuable sources of bioactive substances for various applications in pharmacology, nutraceuticals, and other biotechnological fields.