A method for the simultaneous determination of 12 banned and restricted substances，including enoxacin，ciprofloxacin，ofloxacin，lincomycin，clindamycin，clindamycin phosphate，chloramphenicol，p-nitropheny-2-amino-1-3-propanediol，sulfamethoxazole，metronidazole，tindazole and salicylic acid was developed by reversed phase high performance liquid chromatography（RP-HPLC） with diode array detector（DAD）.The above twelve substances were extracted ultrasonically with methanol-0.1 mol/L hydrochloric acid（1∶1）.The analytes were separated on an ACE C18（250 mm×4.6 mm，5 μm） column by gradient elution with 0.025 mol/L phosphoric acid triethylamine-acetonitrile as mobile phase,and detected at the wavelengths of 210 nm and 277 nm，respectively.The flow rate was set at 1.5 mL/min and the column temperature was 40 ℃. Identification of the 12 banned and restricted substances was achieved by the retention time and the DAD spectra.Quantification analysis was performed by the external standard method.Under the optimal conditions，the detection limits of twelve substances were in the range of 5-50 μg/g.The calibration curves were linear in the range of 25-1 000 mg/L for lincomycin，clindamycin，clindamycin phosphate， and 5-200 mg/L for the rest of analytes，with correlation coefficients more than 0.997.The average recoveries at two spiked levels ranged from 88% to 115% with RSDs of 0.4%-3.0%.The results indicated that the method was simple，sensitive and accurate，and was suitable for the determination of 12 banned and restricted substances in ant-acne cosmetics.