The invention relates to an antibiotic detection method, and in particular, relates to a method for simultaneously detecting various antibiotics in livestock and poultry manure by high performance liquid chromatography; the method comprises the steps: pretreating a livestock and poultry manure sample, to obtain a to-be-detected liquid; performing analyte separation of the to-be-detected liquid by a high performance liquid chromatograph, and calculating peak areas corresponding to separated residual antibiotics; and quantitatively calculating according to a standard curve, to obtain the residual content. The method is applied in detection of 11 kinds of residual antibiotics in the livestock and poultry manure; compared with a high performance liquid chromatography-mass spectrometry coupled technique, the method has the advantages of lower detection cost and high detection efficiency, can simultaneously separate 11 kinds of antibiotics, has analytes without interference of major interferents, and basically does not affect qualitative and quantitative analysis of the analytes.