Simultaneous determination of nine marker substances was established for the quality control of “Byi-Liang-Tang” by high performance iquid chromatography (HPLC). These marker substances include ferulic acid from Cnidii Rhizoma, paeoniflorin from Paeoniae Radix, glycyrrhizin from Glycyrrhizae Radix, cinnamic acid and cinnamaldehyde from Cinnamomi Cortex, puerarin and daidzin from Puerariae Radix, and baicalin and baicalein from Scutellariae Radix. Extracted samples were analyzed with reverse-phase column (Inertsil 5 ODS 2, 4.6 mm i.d. × 250 mm) at 30˚C and eluted with a mixture of 20%, 50% and 90% acetonitrile (adjusted to pH 3.0 with phosphoric acid) aqueous solution in gradients at a flow-rate of 1.0 mL/min, and detected at 230 nm. Relative coefficients of variations of intra- and inter-day analysis were less than 5% and the relative errors were below 2.63% and 1.81%, respectively. All the recoveries were 90~108.67%. This method could be applied for the simultaneous determination of nine marker substances in “Byi-Liang-Tang” and comparisons to commercially preparations.