The invention provides a preparation method of
enzyme-linked immunosorbent
assay (ELISA) kit for detecting
cyproheptadine hydrochloride. The kit is sensitive, accurate and rapid in detection, simple and convenient to operate, high in specificity and suitable for detecting samples in large batch, and comprises an ELISA plate coated with a
cyproheptadine hydrochloride antigen, a
cyproheptadine hydrochloride standard substance, a cyproheptadine hydrochloride
enzyme-labeled
antibody working solution, a substrate solution A, a substrate solution B, a stop solution and a concentrated scrubbing solution. According the principle of the kit, an ELISA reaction is performed through direct
solid-phase competition; an extracted sample and the
enzyme-labeled
antibody working solution are added into a corresponding ELISA hole; after a period of incubation, the ELISA plate is scrubbed, and the substrate solution A and the substrate solution B are added; under the enzymatic action, a blue color appears in the ELISA hole; after the stop solution is added, the
color changes from blue to yellow, and the
color depth has an inversely proportional relationship with the content of cyproheptadine hydrochloride in the standard substance or the sample. The principle can be directly utilized for detecting the content of cyproheptadine hydrochloride in a
swine urine sample.