Abstract: OBJECTIVE To establish an HPLC analytical method for the determination of peoniflorin, astilbin, chlorogenic acid, rosmarinic acid and glycyrrhizic acid in Shaoling Xiaoyin tablets simultaneously. METHODS The chromatographic condition included column Diamonsil C₁₈(250 mm×4.6 mm, 5 μm), acetonitrile(A)-0.1% formic acid(B) as the mobile phase, gradient elution, the column temperature 36 ℃ and detected various wavelengths simultaneously. RESULTS The retention time of peoniflorin, astilbin, chlorogenic acid, rosmarinic acid and glycyrrhizic acid were about 44.2, 57.5, 28.6, 72.3 and 90.6 min, respectively. The regress equation for peoniflorin was y=33 605 228x-124 483, r=0.999 9 and the linear range was 3.25-60.5 μg·mL^-1; astilbin was y=101 354 460x-100 4, r=0.999 9, the linear range was 6.5-130 μg·mL^-1. Chlorogenic acid was y=119 599 014x+22 521, r=1.000 0, the linear range was 0.98-19.6 μg·mL^-1. Rosmarinic acid was y=104 333 261x-365 9, r=0.999 9, the linear was 0.75-15.0 μg·mL^-1. Glycyrrhizinic acid was y=24 700 178x+7 185, r=0.999 9, the linear was 8-160 μg·mL^-1. The average recovery of peoniflorin, astilbin, chlorogenic acid, rosmarinic acid and glycyrrhizic acid was 98.7%, 100.4%, 99.6%, 102.1%, 102.3% and RSD was 2.17%, 0.74%, 2.64%, 1.39%, 1.93%, respectively. CONCLUSION The method is specific, accurate, reproducible, and can exclude interference from other ingredients, so it can be used for quality control.