Abstract: A multi-target method of ultra high performance liquid chromatography-linear ion trap/Orbitrap high resolution mass spectrometry (UPLC-LTQ/Orbitrap MS) was established and validated for the detection of 89 prohibited compounds regulated by (EC) No 1223/2009 on cosmetic products, including antibiotics, hormones, phthalic acid esters (PAEs), pesticide residuals, malachite green and violet. The target compounds were extracted with methanol using ultrasonic extraction. The separation was carried out on a Waters Acquity UPLC BEH C18 column (2.1 mm×100 mm×1.7 μm) with a mobile phase consisting of 5 mmol/L ammonium acetate (0.1% formic acid) and acetonitrile in positive mode and 5 mmol/L ammonium acetate and acetonitrile in negative mode. The retention time, accurate mass of parent ion and relative isotope abundance were used for fast screening, while the generated fragments by collision induced dissociation (CID) were used for comfirmation. With the assistance of Mass Frontier software, fragments generated in MS² collision were deduced and their theoretical assignments were used, which can greatly improve the confirmative accuracy of compounds. The results show that each target has good linearity in its own concentration range and the correlation coefficients is higher than 0.99. The limit of quantification (LOQ) for most compounds are in the range of 5-10 μg/kg except for diphenyl phthalate. For the most compounds, the average recovery at three spiked levels of 1 LOQ,2 LOQ and 10 LOQ range from 60% to 117%, and the relative standard deviation (RSD) is less than 13%. This method is simple, rapid and reliable, which can be used for determination of multi-class prohibited compounds in cosmetics.