Provided is a method capable of separating a target cyclic macrolide compound without using a silver compound even when a compound having a double bond similar to the target cyclic macrolide compound is mixed. A method for separating a cyclic macrolide compound from a mixture comprising the cyclic macrolide compound and at least one analogous compound thereof, wherein the mixture is subjected to silica gel column chromatography using an asymmetry identifying agent-immobilized silica gel.