Abstract: The objective of this study was to establish a HPLC method for analyzing the content of steviol in steviol glycosides.The chromatographic conditions were as follows:column Phenomenex C18 (4.6 × 250 mm,5 μm);mobile phase acetonitrile:water (50:50);flow rate 1.0 mL/min;UV detection wavelength 213 nm;column oven temperature 30 ℃;injection volume 10 μL.The method was linear within the range of 1.046 μg/mL~52.3 μg/mL (r=0.9997),average recovery was 96.60%,RSD of 0.51% (n=6).This method was accurate,precise,repeatable and easy to operate.It can be used for analyzing content of steviol in steviol glycosides.

Key words: steviol, steviol glycosides, HPLC