To compare and analyze the differences of unsaturated fatty acid in Rana chensinensis ovum,Oviductus Ranae,sea cucumber,bird's nest and Cordyceps sinensis,this work simultaneously determined the contents of six common unsaturated fatty acid by using high performance liquid chromatography (HPLC).The results showed that there were significant differences in the contents of unsaturated fatty acid in Rana chensinensis ovum,Oviductus Ranae,sea cucumber,bird's nest and Cordyceps sinensis.Six kinds of unsaturated fatty acids were detected in Rana chensinensis ovum,Oviductus Ranae,including eicosapentaenoic acid (EPA),α-linolenic acid (ALA),docosahexaenoic acid (DHA),arachidonic acid (ARA),linoleic acid (LA) and oleic acid (OA).Five kinds of unsaturated fatty acids,including EPA,DHA,ARA,LA and OA,were detected in sea cucumber samples,which were generally lower than those in Rana chensinensis ovum.Two kinds of unsaturated fatty acids,including LA and OA,were detected in bird's nest,while three kinds of unsaturated fatty acids,including ALA,LA and OA,were detected in Cordyceps sinensis,but they were lower than those in Rana chensinensis ovum,Oviductus Ranae.The results showed that compared with the other four tonics,Rana chensinensis ovum had relatively rich types and higher contents of unsaturated fatty acid,laying a theoretical foundation for the development and utilization of Rana chensinensis ovum.