Squid is one of the export commodities in Indonesia. In general, the use of squid meat, while the ink is only as waste. In fact, Squid ink contain bioactive compound that potential as anti-inflammatory, antihypertensive, anti-diabetic,anti-microbial and anti-malaria agents. The purpose of the study is to determine the types of secondary metabolite compounds contained in n-hexane extract of Loligo sp. ink using maceration method to determine its antibacterial activity against Escherichia coli. The results of secondary metabolite compounds obtained from the n-hexane extract of Loligo sp. ink are alkaloid, saponins, glycosides and phenol. The results of antibacterial test against E. coli using the disc method obtained the average of inhibition zone diameter at the concentration of 4% is 6.3 mm (intermediate), concentration of 8% is 7.83 mm (intermediate), concentration of 16% is 14.5 mm (susceptible) and concentration of 32% is 10.83 mm (intermediate). The antibacterial activity in n-hexane extract of Loligo sp. ink is optimal at the concentration of 16% against E. coli bacteria. © Published under licence by IOP Publishing Ltd.