Anti-Urolithiatic Potential of Hydrocotyle verticillata: Phytochemical Profiling and In Vitro Evaluation of Calcium Oxalate Crystallization Inhibition

Abstract

Urolithiasis, commonly known as kidney stone disease, is a prevalent and recurrent urinary disorder characterized
primarily by the formation of calcium oxalate crystals within the renal system. The increasing incidence of this
condition, coupled with the limitations of conventional treatments such as recurrence, adverse effects and economic
burden, necessitates the exploration of safer and more effective alternative therapies. The present study evaluates the
anti-urolithiatic potential of Hydrocotyle verticillata through pharmacognostic, phytochemical and in vitro
experimental approaches. Preliminary qualitative phytochemical screening of the ethanolic extract revealed the
presence of flavonoids, phenolics, alkaloids, tannins, saponins and glycosides, which are known for their relevance
in urolithiasis management. GC-MS characterization identified major compounds such as benzyl alcohol, glycerin,
D-limonene, dodecanoic acid and 5-hydroxymethylfurfural, which may contribute to antioxidant and crystal-
inhibitory activity. The anti-urolithiatic activity was assessed using in vitro calcium oxalate nucleation and aggregation
inhibition assays, with potassium citrate used as the standard drug. The extract showed strong nucleation inhibition
of 80-90%, compared with 75-85% inhibition produced by potassium citrate. These findings indicate that the extract
can interfere with the early stages of stone formation and crystal aggregation. In conclusion, Hydrocotyle verticillata
exhibits promising anti-urolithiatic potential supported by its phytochemical composition and experimental evidence
of calcium oxalate crystallization inhibition, suggesting its possible use as a natural therapeutic candidate for the
prevention and management of urolithiasis.