Sambiloto (Andrographis paniculata) has been used to treat malaria. In vitro research using parasite or through heme polymerization inhibition using andrografolide and in vivo using infected animal test have been done widely, however, heme polymerization inhibition from extracts with different polarity levels has not been studied yet. The aims of this study were to investigate the heme polymerization inhibition activity and toxicity of sambiloto leaf extracts. Sambiloto leaf extracted with gradually maceration using n-hexane, ethyl acetate, and 70% ethanol respectively. Heme polymerization inhibition activity was used as in vitro antimalarial test. Brine shrimp lethality test (BSLT) was used to determine toxicity of the extracts. Phytochemically screening was done for all extracts qualitatively. The results of this study were n-hexane, ethyl acetate, and 70% ethanol extracts had heme polymerization inhibition activities with IC50 values at 2,196.57; 1,235.54; and 1,157.24 μg/mL respectively. N-hexane, ethyl acetate, and 70% ethanol have LC50 values at 1,155.79; 1,133.89; and 5,229.15 μg/mL respectively. 70% ethanol extract contains alkaloid, flavonoid, steroid/triterpenoid, saponin, and tannin. N-hexane and ethyl acetate extracts has only contains alkaloid and flavonoid. 70% ethanol, ethyl acetate, and n-hexane extracts of sambiloto leaf have ability to inhibit heme polymerization and also non toxic to Artemia salina larvae.