The invention relates to a preparation method of a sea intestine flavor. Various components are mixed parts by weight in a container and are heated to 70 to 85 DEG C, the temperature is kept for 5 to 15 minutes, thus the sea intestine flavor is prepared: 100 shares of sea intestine liquid, 0.3 to 0.6 shares of carnine-5(1)-disodium hydrogen phosphate, 0.3 to 0.6 shares of guanosine-5(1) -disodium hydrogen phosphate, 6 to 12 shares of monosodium glutamate, 4 to 10 shares of common salt, 3 to 10 shares of zymoid monosodium glutamate, 10 to 16 shares of grape sugar, 5 to 7 shares of glycin, 2 to 6 shares of methionine, 5 to 8 shares of proline, 5 to 8 shares of lactamine, and 5 to 8 shares of arginine. The preparation method of the sea intestine liquid is as follows: the sea intestine is made by sea intestine powder added with water in which the weight of the sea intestine powder is 500 to 800 percent. The flavor keeps the original taste of the sea intestine, the preparation technology is simple, the cold storage is not needed, the cost is low, the popularization is easy, and the taste characteristic of the Jiaodong cuisine (a typical cuisine of the Shandong cuisine) is better kept.