Evolvulus alsinoides (Linn.) Linn. is referred as “brain tonic” in Asia and America owing to its activity on the central nervous system. The plant is also found to possess ample of medical activities has been extensively used as traditional medicine in various culture aspects. In the present investigation, the impact of altitude and season of harvest on the phytochemicals and biological activities of E.alsinoides (Linn.) Linn. has been studied. The root, stem and leaves of the plant collected from different altitudes in the Western Ghats of Sathyamangalam, Tamilnadu during the month of July and December, 2016. The primary and secondary metabolites were analysed qualitatively and quantitatively using standard protocols. Minitab 17 Statistical software package was used for Statistical analysis. The altitude and month of plant harvest has an impact on the accumulation of the primary and secondary metabolite in the root, stem and leaves. The leaf samples are found to possess significant antioxidant activities which is further validated by the cytotoxicity activity on SH-SY5Y Neuroblastoma cell line with an IC50 of 119.783 ± 0.06 µg/ml. Isolation of individual molecules and its authentication on the biological activity could give prolific results.