Abstract: OBJECTIVE To analyze and evaluate the fatty acids in Paeonia rockii pollen from Gansu province. METHODS The fatty acids in Paeonia rockii pollen collected from Gansu province and it's wall-broken pollen samples were extracted by ultrasonics method with cyclohexane, then methyl esterificated via BF₃-methanol method, and finally detected by GC-MS-SIM. HP-5MS capillary column(30 m×0.25 mm, 0.25 μm) was adopted, and the heating program was 50 ℃ programmed to 155 ℃ at 8 ℃·min^-1 and kept for 2 min, then increased to 175 ℃ at 10 ℃·min^-1 and kept for 2 min, and raised to 190 ℃ at 1.3 ℃·min^-1 and kept for 8 min, finally increased to 280 ℃ at 6 ℃·min^-1 and kept for 5 min. The sample inlet temperature was set at 260 ℃. The injection volume was 1 μL with splitless mode. High purity helium was used as the carrier gas. RESULTS Paeonia rockii pollen contained 18 kinds of fatty acids (C8-C24). The content of caprylic acid, myristic acid, palmitic acid, linoleic acid, oleic acid and γ-linolenic acid were the more higher content among 18 kinds of fatty acids. The fatty acids in Paeonia rockii pollen were comprised of polyunsaturated fatty acids which takes up 50.45%-73.55% of the total fatty acids. The wall-broken pollen was almost 6-7 times as amounts of fatty acids as pollen. CONCLUSION The method is simple, accurate, which can be used for fatty acids analysis in Paeonia rockii pollen.