The present invention provides a method of screening anti-plasmodial activity of Acriflavin, comprising assessing growth inhibition of plasmodium in vitro in chloroquine susceptible and cloroquine resistant plasmodium by Acriflavin; or measuring in-vivo plasmodium killing ability of Acriflavin; assessing localization of Acriflavin at different stages; and analyzing effect of Acriflavin on gyrase activity wherein said method utilizes Acriflavin in nano-molar range. The present invention relates to potency of Acriflavin (Acriflavin) as an anti-malarial agent both in vitro parasite culture as well as in vivo. More specifically, the invention relates to a method o determining anti-plasmodial activity, Acriflavin as potent anti-malarial agent and also relates to composition(s) comprising Acriflavin.