A pre-irradiation separation procedure, for gallium determination in biological and environmental materials by neutron activation analysis (NAA) is reported. The proposed method, based on ion-exchange chromatography, allows to eliminate the radiation risk, while taking the advantages of high sensitivity of NAA. The single comparator method is employed for gallium quantitative evaluation. Since no standard reference materials with certified gallium contents were available, the reliability of the proposed method was extensively investigated by various approaches. Four biological and one environmental standard reference materials, with gallium contents ranging from 54 g/g-3 ng/g, have been analyzed.