Isolation, anticancer activity test, and identification of the toxic isolate from ethanol extract of the sponge Hyrtios erecta taken from Pari Island beach (Jakarta) has conducted. Extraction of the sponges was carried out by 70% ethanol at room temperature. Partition and purification of the compounds were done by column chromatography with the stationary phase of silica gel and the mobile phase of n-hexane-chloroform (2:8). Toxicity screening test was done based on Bhrine Shrimp Lethality Test (BSLT). In vitro anticancer activity test of the isolate was carried out using HeLa cell line. Identification of the compounds was performed by Gas chromatography-mass spectroscopy (GC-MS). Based on the results, it was found that the toxic isolate of H. erecta sponges has anticancer activity with IC50 of 30,497 ppm. Four compounds was detected from the anticancer isolate i.e: 4-nonylphenol; dibutyl phthalate; hexanedioic acid bis(2- ethylhexyl) ester; and cholesterol.