Objective To develop an RP-HPLC method for simultaneous determination of mangiferin, tectoridin, iridin, tectorigenin, iristectorigenin B, iristectorigenin A, irigenin, dichotomin and irisflorentin in Belamcandae Rhizoma.Methods Separation was carried out on an LeapsilTM C18 column (100 mm×2.1 mm, 3 μm) with an isocratic mobile phase consisting of acetonotrile and formic acid at a flow rate of 0.5 mL/min; The detection wavelength was set at 265 nm; the column temperature was 40℃.Results The linear ranges of mangiferin, tectoridin, iridin, tectorigenin, iristectorigenin B, iristectorigenin A, irigenin, dichotomin and irisflorentin were 0.214 0– 2.568 μg (r=0.999 5), 0.437 0–5.244 μg (r=0.999 3), 0.460 0–5.520 μg (r=0.999 9), 0.078 40–0.940 8 μg (r=0.999 6), 0.138 0–1.656 μg (r=0.999 3), 0.051 00–0.612 0 μg (r=0.997 5), 0.113 0–1.356 μg (r=0.999 9), 0.051 63–0.619 6 μg (r=0.999 8) and 0.151 0–1.812 μg (r=0.999 9), respectively. The average recoveries were 97.73%, 96.81%, 97.78%, 97.55%, 96.86%, 98.60%, 97.77%, 98.04% and 97.89%, respectively; the relative standard deviations were 0.70%, 1.1%, 2.3%, 2.1%, 1.3%, 1.4%, 2.3%, 1.6% and 1.9%, respectively. This method was used to determine the contents of nine active ingrients in 5 batches of Belamcandae Rhizoma.Conclusion The method is accurate and reliable, which can be used for the quality control of Belamcandae Rhizoma.