The invention relates to β-carotene microcapsules rich in sardine oil, a preparation method and application thereof. In the present invention, denatured starch, chitosan, trehalose, sucrose, and monoglyceride succinate are sequentially added to water, stirred while adding, mixed evenly, and the solution is kept at a certain temperature to obtain the water phase, β-carotene crystals, Add lipophilic emulsifier Tween and sardine oil into the oil phase kettle, stir and dissolve to obtain the oil phase, slowly add the oil phase into the water phase, and make a microemulsion by shearing, and the microemulsion is prepared by concentrating, freeze-drying, grinding, etc. Microencapsulated beta-carotene enriched with sardine oil. Compared with the prior art, the obtained β-carotene microcapsules of the present invention not only improve the stability of β-carotene to factors such as light, heat and humidity, but also are rich in sardine oil in the microcapsules, which can combine with β-carotene Play a synergistic effect and improve the health care performance of microcapsules.