The invention relates to a composite preparation for improving fish body transportation stress capability as well as a preparation method and application thereof. The composite preparation for improving fish body transportation stress capability comprises the following ingredients in parts by weight: 40-60 parts of tryptophan, 4-8 parts of nucleotide, 20-30 parts of Schisandra chinensis, 0.5-1.5 parts of tea polyphenol, 1-3 parts of vitamin E and 14.5-17.5 parts of carrier. The preparation method comprises the following steps: preparing the ingredients in parts by weight; fully and evenly mixing all the ingredients; and smashing to prepare the composite preparation of dry powder of 100-120 meshes. The composite preparation is used in a culture water body before the fish body is transported and in a transportation water body in the transportation process, and the use concentration is 10-20ppm. The preparation method has the advantages of low energy consumption, simple technology, low production cost and the like, and the composite preparation has the functions of improving fish body transportation stress resistance capability and increasing the survival rate.