Biopharmaceuticals require exceptional purity, however, purification process is a challenging task. To achieve the required levels of purity, several chromatographic steps need to be implemented. Liquid chromatography is still main technique of choice used for purification of biological substances. Quality of packed chromatographic column plays a significant role in the performance of the chromatographic step. In the present study we determined the impact of packing of chromatographic columns on product quality. For this purpose, we packed ten columns differently in order to obtain various quality of packed chromatographic columns. After the columns were packed, we tested them before and after each separation using a pulse test in order to assess their quality defined by three measures: number of plates, height equivalent to the theoretical plate and asymmetry. We intentionally packed columns that differed in all parameters and deviated from recommended values. After column preparation, a separation using the hydrophobic interaction chromatography was carried out on each of packed columns. The analysis of the eluates showed that the content of impurities was consistent between individual chromatographic columns. Taking into account the uncertainty of the analysis, we concluded, that all of the packed columns are equivalent concerning the purification of product; moreover, each of the columns purified the product to proper levels of purity.