Abstract: The present research reports the results of the phytochemical studies carried out to identify the diagnostic features of the E. alba. A high performance thin layer chromatographic method was standardized to determine the wedelolactone content in whole plant of E. alba. Methanolic extracts of samples from three different sources were used for analysis. The mean assay of wedelolactone was of range 0.481-0.702 mg gG1 of drug powder. Radical scavenging activity of methanolic extract was evaluated by DPPH assay method, superoxide radical scavenging activity in riboflavin/light/NBT system and nitric oxide radical scavenging activity in sodium nitroprusside/Griess reagent system. The assay results indicate that the DPPH, superoxide and nitric oxide scavenging activity were intense (19.25, 39.25 and 58.26 μg mL1, respectively). The phytochemical features identified in the present study can be used as identification markers of this important analgesic agent.