In previous study, in vitro antiplasmodial activity fractions isolated from methanol extract of E. longifolia, Jack. have been evaluated. Among 5 isolates evaluated from the study, isolate 4 showed high in vitro antiplasmodial activity. However, which stage specificity of the isolates on P. falciparum cycles has not been evaluated. This study was intended to evaluate the stage specificity of the isolate on P. falciparum cycles. The study was conducted by observing the percentage of each stages of P. falciparum microscopically after 8, 16, 24, 32, 40, 48, 56, 64, and 72 hours incubation periods with 3 various concentration of isolate 4 compared with control. The result showed that isolate 4 of E. longifolia root methanol soluble fractions most potent at trophozoites stages of P. falciparum.