The invention provides a purification method of tylvalosin. The method is characterized by comprising the following steps: (1), dissolving a tylvalosin crude product with a solvent, adding petroleum ether to perform crystallization, and performing filtering and drying, so as to obtain tylvalosin crystal, wherein the solvent is one or more of ethyl acetate, butyl acetate and tetrahydrofuran; and (2), adding the tylvalosin crystal into isopropanol, so as to be dissolved, dropwise adding petroleum ether to perform crystallization again, and performing filtering and drying, so as to obtain high-purity tylvalosin. The method is simple to operate, mild in condition, relatively high in yield, replaces column chromatography isolation in a crystallization mode, and greatly saves time and eluents, so that the purity of obtained tylvalosin can be 95% or higher.