The invention discloses fat globule membrane powder, a preparation method of the fat globule membrane powder and application of the fat globule membrane powder. The preparation method comprises the following steps of adjusting the pH value of raw materials to be 7-8, performing separation and concentration by utilization of a membrane material with pore diameter of 0.1-1.5 microns, filtering, washing, collecting a concentrated solution in an intercepted section, freezing and drying. The raw materials are fluid milk products containing the fat globule membrane powder. The fat globule membrane powder is prepared by utilization of a membrane separation method, the preparation method is simple, the separation efficiency is high, the industrialized production is conveniently performed, the content of a fat globule membrane ingredient is high, and the damage to a fat globule membrane structure as well as the oxidization of lipid components in the fat globule membrane are both avoided. The fat globule membrane powder is applied to UHT (Ultra High Temperature) single cream production; the prepared single cream is rich in natural phospholipid ingredients, can soften blood vessels, is beneficial to human health and has higher nutritional properties; the fat globule membrane powder has a better stability, has better application performance before the expiration date, has a shelf life of 6-9 months, cannot cause the phenomena such as fat floating, and has a better storage stability and application prospects.