In this paper, three higher content of flavonol glycosides from the stems and leaves of Tribulus terrestris L. were determined by reversed phase HPLC. The results showed that there was good liner relationship between the peak areas and the sample concentration at the ranges of 0.011 2~0.280 0 μg (r=0.999 6), 0.064 8~2.592 0 μg (r=0.999 8) and 0.018 4~0.460 0 μg (r=0.999 8) for isorhamnetin-3-O-β-D-gentiobioside-7-O-β-D-glucoside, quercetin-3-O-β-D-gentiobioside and isorhamnetin-3-O-β-D-gentiobioside respectively. The verage recovery rates (n=9) of three flavonol glycosides compounds were 100.15%(RSD=1.32%), 100.02% (RSD=1.14%), 99.77% (RSD=1.16%), respectively. This method is considered to be simple, accurate, stable, good precision and reproducibility, which is available for the quality control and evaluation of T. terrestris L.