Objective：To establish a method for determining the contents of Stellettin B and Stellettin D in total isomalabaricanes in different batches of Stelletta tenuis Lindgren. Methods： A high pressure liquid chromatography （HPLC） method was applied. The column YMC Pack-SIL （250 mm×4.6 mm, 5 μm） was used at room temperature, and the mobile phase was nhexane ： ethyl acetate （75 ： 25） with a flow rate at 1.0 ml · min^-1 for 30 min. UV detector wave length was at 360 nm. Resuits： The regression equation were：Y=52 181 633 X＋957 495（r=0. 996 9）for Stellettin B and Y=62 636 828 X-451 022 （r=0. 998 0） for Stellettin D,with the linear range being 2.4-30μg and 2.0-32μg, correspondingly. The RSD values for accuracy test, stability test and repeatability test were 3.07%-4.76%. The average contents of Stellettin B and Stellettin D in total isomalabaricanes in different batches of Stelletta tenuis Lindgren were 59. 93% and 30. 98%, respectively. Conclusion： This HPLC method provides a fast, accurate and effective way to determine the active contents in total isomalabaricanes from Stelletta tenuis Lindgren.