With introduction of a diisopropyloxy phosphoryl group into the N terminal of amino acids, it was found that proton affinity (PA) of amino acid was enhanced in mass spectrometry. Density functional theory calculations showed that the energy for protonation of DIPP-amino acid is lower than that of amino acid, which means PA of DIPP-AA is higher than that of corresponding amino acid. These results, coincident with our empirical results, offer a useful interpretation of experimental observations.