The performance of several alkyl-bonded alumina-based stationary phases was evaluated by comparing the separation of synthetic octapeptide and polypeptide mixtures and tryptic digests of larger proteins. These phases were of differing pore diameter, alkyl chain length modification and particle shape and size. The separations were compared to standard silica phases. The narrow pore octadecyl bonded alumina phase outperformed the other alumina and silica phases in terms of separation efficiency and mobile phase resistance. Superior performance is attributed to the enhanced solute mass transfer properties and the unique morphology of the microplatelet alumina particles. The mechanism of separation gradually changes with increasing size of the peptide.