Porous anodic alumina (PAA) membranes were used as a nanometre-order diameter column for chromatography chip with acetonitrile-water mobile phase using gradient elution. In a chromatogram, 9-anthracenemethanol (AM), 9-anthracenecarboxylic acid (AC), and dansyl-glycine (DG) showed different retention times, indicating that PAA membranes are applicable to stationary phase for chromatography chip.