Resistance is inherited and has proved to be the biggest single barrier to successful chemical control of insect vectors. The continuity of along time period insecticide usage can produce mosquitoes resistance. Resistance to insecticide as a results from three main mechanism : 1) insecticide penetration is reduce, 2) the insecticides is more efficiently metabolized by esterases, mixed function oxidases, or glutathione transferase enzyme and, 3) the target of the insecticide is modified (insensitive acetylcholinesterase). The objectives of this study was to determine the potency of malaria vector from East Java Province to be resistant to organophosphate, carbamate and pyrethroid insecticides. The research methods used were biochemical assays (microplate assays) for elevated esterase and insensitive acetylcholinesterase. The esterase activity and insensitive acetylcholinesterase were measured at 450 nm and 405 nm with a Dytech Elisa plate reader. Biochemical assays indicated that susceptibility (resistant or tolerance), of the malaria vector collected from East Java Province natural population against insecticide were mostly decreased, although there were different level and mechanism occurs. Microplate enzymatic assay on individual Anopheles sundaicus collected from Banyuwangi, Pacitan, Jember, Malang Regency revealed that 31,25 %, 12,5 %, 45,2 % and 15,25 % population were resistant respectively due to elevated esterase activity mechanism. Base on the susceptibility test which was held using WHO method (as a cross-check) An. sundaicus from Teleng village, Pacitan Regency was proven to have a double resistance agains Malathion 0,5 % (cause mortality 28 %) and Bendiocarb 0,1 % (cause mortality 66 %). The percentage resistance of Anopheles aconitus population collected from Pacitan and Trenggalek Regency were 35,42 % and 29,17 % population respectively due to elevated esterase activity mechanism. There was no evidence of an altered acetylcholinesterase (insensitive acetylcholinesterase) mechanism of the malaria vector population in East Java.