The characteristics of Cr3+andCr6+ ion separation in solution through biosorption process using green algal biomass Spirogyrasubsalsa with batch systems have been investigated. The study began with aqualitative analysis of functional groups in biomass using FTIR instrument, then followed by a study of the characteristics of influences by several variables, such as: the initial pH of the solution,the size of biosorben particles, stirring speed, the effect of heating the biosorben, the rate of absorption, and the effect of metal ion concentration in solution on the absorption capacity of algal biomass. Based on FTIR spectroscopy spectra gave a conclusion that biomass of green algae S.subsalsa contains carboxylate groups, amine, amide, amino, carbonyl and hydroxyl, in addition to silicon, sulfur, and phosphorus compounds. The results showed that the biosorption capacity was strongly influenced by pH, contact time, and initial concentration ofthe solution. The optimum biosorption of Cr3+cation occurred at pH 4.0 while that of Cr6+ions occurred at pH 2.0 and then decreased with the increasing pH of solution. The calculation of Langmuir isotherm equation showed that the maximum absorption capacity of algal biomass S.subsalsa for Cr3+and Cr6+ ion respectively was 1.82mg (0.035 mmol) and 1.51 mg (0.029 mmol) pergram of dry biomass. The kinetics of biosorption took place relatively quick, in which during the 30minutes time interval, each ion was absorbed approximately 95.7%; and 86.5%. The absorptive capacity of biomass was also influenced by stirring speed, while the size of particles and heating biosorben gave lessinfluence to the absorption of biomass.