The interaction between cephalosporins and the dextran matrix of Sephadex gel in gel filtration chromatography has been thoroughly studied. Twelve cephalosporins with specific structures were examined under different chromatographic conditions, including 12 different mobile phases comprising inorganic or organic compounds of different charge or/and density of electrons on their negative ions, different types of Sephadex gel (Sephadex G-10 and Sephadex G-50) and different flow rates. It was found that the more negative the charge or/and density of electrons on the negative ions of buffer components, the more was the adsorption of cephalosporins on the solid phase; this indicated tht the mobile phase played an important rôle in gel filtration chromatography for cephalosporins. By choice of suitable chromatographic conditions, optimum separation of high molecular weight impurities from cephalosporins could be achieved. The novel method could be used as a routine method for the quality control of cephalosporin preparations.