A new RP-HPLC method was developed for the simultaneous determination of 5 active constituents, including loganic acid, swertiamarin, gentiopicroside, sweroside, isoorientin in aerial part of Gentiana straminea. Analysis was achieved on a Hypersil ODS analytical column (4.6 mm x 250 mm, 5 microm) eluted with methanol and water containing 0.02% phosphoric acid in gradient elution. The flow rate was 1 mL x min(-1), the column temperature was set at 35 degrees C and detection wavelength was set at 254 nm. The results showed that 5 active components were separated well and showed good linearity. The corelation coefficints and concentration ranges of the calibration curves were as follow: r = 0.999 9, 0.364-2.18 microg for loganic acid. r = 0.999 9, 0.275-1.65 (microg for swertiamarin. r = 0.999 9, 0.614-3.68 microg for gentiopicroside. r = 0.999 9, 0.065 6-0.394 microg for sweroside. r = 0.999 9, 0.089 9-0.539 microg for isoorientin. The contents of loganic acid and gentiopicroside far exceed that of the other 3 active components, and made up 60% in the total contents. The developed method was proved to be rapid, sensitive, accurate, credible and repeatable. It can be applied to quality control of aerial part of Tibetan medicine Gentiana straminea.