Defatted pine nut meal was used as material, four single factors of NaCl extraction solvent (0.5%, 1.0%, 1.5%, 2.0%, 2.5% and 3.0%), ratio of material to solvent (1∶10, 1∶15, 1∶20, 1∶25, 1∶30, and 1∶35), extraction time(30, 60, 90, 120, 150, and 180 min) and extraction temperature (40, 45, 50, 55, 60, and 65 ℃) were designed to extract globulin by the method of Osborne and to improve extraction process by response surface method and to analyze relative molecular mass of globulin by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS–PAGE). The result showed that the optimum extraction conditions were 2.5% of NaCl, extraction at 56 ℃ for 150 min with ratio of material to solvent 1∶15, the extraction ratio of globulin was up to 15.32%, and the relative molecular mass of the proteins were 41 320, 27 520, 21 690, 16 310, 15 180.