Objective To simultaneously determine the contents of six marker components, including benzoylmesaconine,benzoylaconine, benzoylhyaconine, isoliquiritigenin, glycyrrhizic acid, and 6-gingerol, in Sini decoction by high performance liquid chromatography (HPLC). Methods The HPLC condition was as follows: column: Waters Terra C₁₈ (3.0 mm×100 mm,3.5 μm); the detective wavelength was set at 370 nm for isoliquiritigenin and 235 nm for the other five components. Mobile phase: A was 95% acetonitrile+5% H₂O (5 mmol/L ammonium acetate), B was 0.1% formic acid aqueous solution (5 mmol/L ammonium acetate), with gradient elution, the gradient of A phase:25%-35% (0-5 min),35%-50% (5-15 min), 50%-85% (15-20 min); flow speed: 0.5 mL/min; temperature of column: 25℃; injection volume:5 μL. Sini decoction was obtained by using the traditional decoction preparation method. Results Benzoylmesaconine, benzoylaconine, benzoylhyaconine, isoliquiritigenin, glycyrrhizic acid and 6-gingerol were separated at baseline within 15 min, showing good linearity (r>0.999 0) between (5.600-112.0) μg/mL, (6.560-131.2) μg/mL, (6.130-122.6) μg/mL, (4.590-91.8) μg/mL, (31.00-620.0) μg/mL, and (4.920-98.4) μg/mL, respectively. The results of intra-day and inter-day precisions were at normal range (RSD<5%), with the recovery rates (n=6) being 101.07% (RSD=1.3%), 98.72% (RSD=1.1%), 101.57% (RSD=1.8%), 101.71% (RSD=3.6%), 102.12% (RSD=2.3%), and 99.58% (RSD=3.8%), respectively. Conclusion The present method is rapid, simple, and accurate and it can be used to determine the above 6 components in Sini decoction.