The invention belongs to the technical field of preparation of release coatings and discloses a water-based heat transfer printing release coating, and a preparation method and application thereof. The water-based heat transfer printing release coating comprises the following ingredients by mass percent: 40-60 percent of water-based resin, 8-15 percent of a coalescing agent, 0.3-2 percent of a scratch-resistant anti-wear agent, 0.05-0.2 percent of a foam killer, 0.2-0.6 percent of other additives and the balance of water, wherein the water-based resin is at least one of polyurethane resin, polyacrylic acid resin, polycarbonate, polymide resin, polyethersulfone resin, epoxy resin, thermoplastic polyester resin, a styrene-maleic anhydride copolymer and phenolic resin. According to the invention, application of the water-based formula to preparation of the water-based heat transfer printing release coating is realized, the material prescription is environment-friendly, and the preparation process is safe and environment-friendly. Compared with a conventional solvent product, the water-based heat transfer printing release coating is better in heat resistance, high in glossiness and better in printing ink adhesion, effectively enhances the printing ink protecting function and can be kept colorful in a long term.