The invention relates to an oligopeptide phase reversal chromatography stationary phase and a preparation method thereof. The preparation method adopts 'link chemistry' as a bonding reaction method to bond oligopeptide molecules and comprises the following steps: firstly, introducing terminal alkynyl on a silica gel surface; and then, taking methanol, acetonitrile, dimethyl sulfoxide or a mixture of the solvents as a reaction solvent to bond oligopeptide modified with azide groups on the silica gel surface, thereby obtaining the oligopeptide phase reversal chromatography stationary phase. The oligopeptide phase reversal chromatography stationary phase has orderly structure and single reaction site; moreover, the structure of the stationary phase introduces triazole rings, urea bonds, benzene rings, peptide bonds and ester groups to generate multiple kinds of acting force, such as hydrogen-bonding action, electrostatic interaction, Pi-Pi action, dipole-dipole interaction, hydrophobic interaction and the like, thereby realizing orthogonal separation on complex compositions in a two-dimensional structure constructed with C18; and 'link chemistry' is adopted as the bonding reaction method to realize immobilization of high selectivity and high conversation efficiency under moderate conditions.