Separation of Peptides in NP-HPLC By HCI Program

초록

The optimization of mobile phase condition for 4 biological-active peptides was demonstrated by HCI program. The binary system of water with 0.1% TFA and acetonitrile with 0.1% TFA was used. The elution profiles were calculated by the plate theory based on the linear (ln k = A+BF) and quadratic (ln k = A+BF+CF2) equations of retention factor, and F was the volume percent of acetonitrile. We modified the plate theory to calculate elution profile in both isocratic and gradient mode. By the calculated results, the first mobile phase composition was water with 0.1% TFA/acetonitrile with 0.1% TFA, 81/19 vol.%, then after 7 min to 8 min, the second composition of mobile phase was linear-changed to 79/21 vol.%, and finally after 8 min, it was kept at the isocratic mode. The agreements between the experimental data and the calculated values were relatively good.