Plantago major L. is a plant of the Plantaginaceae family which has been investigated on diethyl ether extract generate active spot as an antibacterial. The aim of the investigation was to identify the antibacterial compound from diethyl ether extract of daun sendok. as much 500 gram material was extracted maceration method was obtained 50 gram methanol extract, continued by partition with diethyl ether obtained 19.5 gram dried extract, the extract diethyl ether which isolated by suction column chromatography obtained 3 fractions (F1, F2 and F3). The third fraction (F3) was isolated by column chromatography obtained 2 fractions (3-1 and 3-2). The 3-1 active fraction (inhibit the Escherichia coli) isolated by preparative-TLC obtained 2 compounds. Compound 1 was active, single spot and visualization with Lieberman-Buchard and vanillin-sulfate acid. It have a steroid compound.