The invention relates to a method for determining residue of quinolones in a cultivation/slaughter environment, which is realized through the following steps: a sample of water body or soil is collected from the cultivation/slaughter environment for cattle, sheep, pigs, chickens, fishes or shrimps, is treated through an immunoaffinity column, passes through an organic microporous filter membrane of 0.22 micrometer, and is analyzed and tested by means of liquid phase-mass spectrum/ mass spectrum, so that a chromatographic peak of the sample can be determined; then the determined chromatographic peak is contrasted with standard chromatographic peaks of all known quinolones, so that the specific type of a tested drug can be accurately determined. The method is used for determining residue of quinolones in the water body sample or the soil sample of the cultivation/slaughter environment, the pretreatment process of the sample is fast, simple and convenient, and the testing result of the treated sample is reliable and sensitive.