A HPLC chromatography for the determination of erythromycin A oxime and relative compounds was studied, and the effect of chromatography systems including a HITACHI L-7100, a Shimadzu LC-6A, a Waters 474 and relative columns was analyzed. It was revealed that different HPLC apparatus and columns have obvious impact on the peak separation and retention time under the general chromatographic condition. The suitable chromatographic conditions for several different chromatography systems were summarized with good linear relationship, which is very significant to the quality control of erythromycin A oxime and relative compounds.