The retention behavior of five cephalosporins was investigated by thin-laye
r reversed-phase ion-pair chromatography (RP-IPC). Optimization of the rete
ntion and selectivity of these compounds was carried out by changing the pH
, the concentration of the ion-pairing counter-ion, and the concentration o
f the organic solvent in the aqueous mobile phase. The effects of various c
ationic and anionic pairing reagents in the mobile phase and the stationary
phase were investigated. The use of stepwise gradient elution improved the
spot shape and the selectivity of the separation.