Abstract: OBJECTIVE To establish a HPLC method for the simultaneous content determination of (R,S)-goitrin and clemastanin B in banlangen granules. METHODS The analysis methanol was performed on a Agilent ZORBAX SB-C₁₈ column(250 mm×4.6 mm, 5 µm), the mobile phase was methanol-water flowing of gradient elution, the flow rate was 0.8 mL·min^-1, the injection volume was 10 µL, the column temperature was 30℃, and the detection wavelength was 230 nm. RESULTS (R,S)-goitrin and clemastanin B showed good linear relationships within their own ranges of 2.39-47.80 µg·mL^-1 (r=1.000 0) and 3.18-63.60 µg·mL^-1(r=0.999 9). The average recoveries were 100.90%(RSD=1.56%) and 99.48%(RSD=1.08%). CONCLUSION This method is simple, accurate, specific and reproducible, can provide a basis for raising banlangen granules quality standards.