Ferrous sulfate-loaded gelatin microspheres were prepared by premix membrane emulsification, using soybean oil with food grade as oil phase and glucose as solidification agent, the effects of preparation conditions on microspheres morphology and size distribution examined, and the optimal conditions of gelatin microspheres obtained. The results showed that under the optimal concentrations of gelatin solution and emulsifier at 0.200 and 0.07 g/mL respectively, homogenizing rate 10000 r/min, and solidification time 20 min, the gelatin microspheres loading ferrous sulfate were successfully prepared. The microspheres were spherical and uniform in size, and their mean size was 50 μm and encapsulation efficiency of ferrous sulfate 44.12%, containing 60.8% ferrous ion.