Analysis of cephalosporins by hydrophilic interaction chromatography

A simple hydrophilic interaction chromatography (HILIC) method was developed to analyze seven cephalosporins. These seven cephalosporins could be separated well on the Click β-CD column and Atlantis HILIC Silica column. The effects of buffer concentration and pH on the retention under HILIC mode were studied. Except cefepime hydrochloride (4), the retention of other six cephalosporins increased with increasing buffer concentration, while decreased with increasing pH. Different separation selectivities could be observed on the Click β-CD column and Atlantis HILIC Silica column, and changing pH also resulted in the changing of separation selectivity. The separations of cephalosporins by HILIC and reversed-phase high performance liquid chromatography (RP-HPLC) were compared, and the two separation modes had good orthogonality. In addition, cefotaxime sodium (1) and its degradation were separated well on the Click β-CD column, which indicated that the Click β-CD column by HILIC can be used for studying the stability of cephalosporins.