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Andrographis paniculata Ness is one of the plants that is under explored and could contain potentially active substances to serve as an antimalarial. Structure investigation of major compounds that have responsibility for the antimalarial activity of Andrographis paniculata Ness’s n-hexane extract is very important so that it is known whether the antimalarial activity is synergistic or from the major compounds. Menthol and dioctyladipate as major component from n-hexane extract of Andrographis paniculata Ness have been succesfully isolated using simple and inexpensive methods. The solvent system used are n-hexane : ethyl acetate (10:1) and n-hexane: chloroform (10:3) consecutively either for column or preparative thin layer chromatography. FT-IR, 1H-NMR and GC-MS were used to determine the structure of compounds. The activity of n-hexane extract, menthol and dioctyladipate related to the inhibition of heme polymerization have been done comprehensively. The inhibition of heme polymerization activity of isolate compounds and n-hexane extract are classified to a good level (dioctyl adipate IC50 = 1.15 ± 0.41 mg/mL, menthol IC50 = 0.31 ± 0,01 mg/mL, and n-hexane extract IC50 = 0.07 ± 0.03 mg/mL) and potentially as antimalarial. From the IC50, the antimalarial activity of Andrographis paniculata Ness’s n-hexane extract is working synergistically, not by the major compounds.