Hop extract is a complex mixture of alpha and beta acids, essential oils, tannins, and waxes. Components of the hop extract were separated by mixed-mode HPLC on a Primesep SB column. Compounds on this column are retained by combination of reversed-phase and anion-exchange mechanism. At higher pH (3-5), alpha and beta acids are ionized. Retention time can be controlled by amount of acetonitrile, buffer pH, and buffer concentration. Primesep SB column is fully ionized within operational range of the column.