Introduction: The Cayratiaauriculata has been used as a folk medicine to treat various pathophysiological conditions. Aim: In the present study, we evaluated the presence of major phytochemicals, cytotoxicity and acute toxicity effect of ethanolic extract of Cayratiaauriculata leaves. Methodology: The phytochemical screening was carried out using chemical methods; gas chromatography-mass spectrometry (GC-MS) analysis was performed to identify the individual phytoconstituents present in it; cytotoxicity assay was performed in A549 cells and acute toxicity test was performed in the zebrafish model. Results: The results of the qualitative analysis revealed the presence of flavonoids, phenols, coumarin, saponins, tannins, terpenoids, steroids and glycosides. C. auriculatawas found to contain a significant amount of total flavonoid and phenol contents in quantitative analysis. Fifteen different phytoconstituents were expressed in GCMS analysis. In the acute toxicity test, the methanolic extract of C.auriculatadid does not cause mortality or any clinical signs of toxicity in zebrafish to the maximum concentration of 100 mg/L.Therefore the LC50 value of extract was found to be ?100 mg/L. Hence it can be considered safe. Conclusion: This paper will help in considering C. auriculate or further pharmacological studies in future.