Euphorbia hirta is a medicinal plant having rich source of secondary metabolites. Flavonoids are shown to have antioxidative activity, free radical scavenging capacity, coronary heart disease prevention, hepatoprotective, antiinflammatory, and anticancer activities, while some flavonoids exhibit potential antiviral activities. The present study was carried out on the qualitative and quantitative estimation of flavonoid content in suspension culture. The results showed that there was an increase in the flavonoid content as the days progressed.