The invention discloses a composition for detecting the content of an ephedra source component in glue traditional Chinese medicines and a compound preparation of the glue traditional Chinese medicines, a kit and a detection method of the kit. The composition comprises polypeptide and a reference substance detection matrix, wherein the amino acid sequence of the polypeptide is Gly-Glu-Pro-Gly-Pro-Ala-Gly-Ser-Val-Gly-Pro-VaL-Gly-Ala-Val-Gly-Pro-Arg, and the reference substance detection matrix is fishbone glue. The polypeptide is specific to a fishbone; whether the ephedra source component is contained or not is judged by detecting the polypeptide; and the accurate detection of the content of the ephedra source component is achieved by adding the reference substance detection matrix. The method has the advantages of accurate content detection and fast operation, and has wide application prospect in the field of quality monitoring of the glue traditional Chinese medicines and a compound preparation product thereof.