The aim was to establish a method for the simultaneous determination of 2kinds of cholesterol oxidation products (COPs:7-ketocholesterol,cholesta-4,6-dien-3-one) by HPLC.MethodsThe mixture of samples and Florisil packing were grinded, then packed to a syringe column. The extraction and purification of the samples were carried out with one-step using acetonitrile as eluting reagent. With anhydrous ethanol as mobile phase, the two components were determined by liquid chromatography under 240-285nm. ResultsThere was a good linearity in the range of 0.25-100μg/ml and the correlation coefficients were greater than 0.9993. The detection limits for 2COPs were 0.015and 0.045μg/g, respectively. Under the optimal conditions, the recovery rates of the two target analytes were between 74.1%-97.0% and the relative standard deviations (RSDs) were no more than 5.57%.ConclusionThis method is suitable for the determination of cholesterol oxides in chicken liver.