The invention relates to a method for simultaneously screening multiples categories of drug residues in fish by using ultra performance liquid chromatography-quadrupole rod time-of-flight mass spectrometry. The multiples categories of drugs screened simultaneously comprise enrofloxacin, danofloxacin, tetracycline, oxytetracycline , chlortetracycline, doxycycline, chloramphenicol, thiamphenicol, florfenicol, sulfamethoxazole, sulfamethoxazole, sulfathiazole, sulfadiazine, sulfadoxine, sulfisoxazole, sulfaphenirazole, sulfacetamide, sulfamethazine, azithromycin, tilmicosin, medimycin, roxithromycin, acetylspiramycin, doramectin, sudan I, sudan II, sudan III, sudan IV and rhodamine B. By optimizing the parameters of a d-SPE pretreatment method, all target drugs can obtain good recovery rates.29 kinds of target drugs are simultaneously screened by an optimized chromatographic mass spectrometry condition, thereby achieving simultaneous screening of common multiples categories of drug residues in fish. The method has a good application prospect in the field of fish food.