A deformable gel-packed chromatographic column was used to separate as-synthesized graphite oxide with different sizes. The synthesized gel (56 mu m) was deformed by pressure of the fluid flow and the gaps in the gels showed a range of sizes. A suspension of graphene oxide (0.1 g/L, 10 mL) was injected, and graphene oxide in the elution had a size at 0.56 mu m and 0.14 mu m, whereas in half upper and bottom domain of the gel layer graphene oxide had a size at 33 mu m and 2.9 mu m, respectively, demonstrating that graphene oxide suspension was separated by size through gel layer.