Specific and sustained release of nutrients from capsules to the gastrointestinal tract has attracted many attentions in the field of food and drug delivery. In this work, we reported a monoaxial dispersion electrospraying-ionotropic gelation technique to prepare multicore millimeter-sized spherical capsules for specific and sustained release of fish oil. The spherical capsules had diameters from 2.05 mm to 0.35 mm with the increased applied voltages. The capsules consisted of uniform (at applied voltages of ≤ 10 kV) or nonuniform (at applied voltages of > 10 kV) multicores. The obtained capsules had reasonable loading ratios (9.7%−6.3%) due to the multicore structure. In addition, the obtained capsules had specific and sustained release behaviors of fish oil into the small intestinal phase of in vitro gastrointestinal tract and small intestinal tract models. The simple monoaxial dispersion electrospraying-ionotropic gelatin technique does not involve complicated preparation formulations and polymer modification, which makes the technique has a potential application prospect for the fish oil preparations and the encapsulation of functional active substances in the field of food and drug industries.