Abstract: A method of simultaneous determination of 5 active chemical constituents (adenosine, ginsenoside Rb₁, β-sitosterol, vanillic acid and liquiritigenin) in Polygonatum Sibiricum Red. at different growth stages was proposed by high performance liquid chromatography with dual wavelengths. The sample (5.000 0 g) was extracted ultrasonically for 30 min with 40 mL of methanol (70+30) solution. The extract was separated by HPLC with dual wavelengths (λ₁=252 nm, λ₂=280 nm) using Diamonsil C₁₈ column as the stationary phase, and a mixture of 0.1% (φ) formic acid solution and 0.1% (φ) formic acid-acetonitrile solution in different volume ratios as the mobile phase for gradient elution. Under the optimized test conditions, linear relationships between values of peak area and mass concentration of 5 active chemical constituents were found in the range of 0.5-100 mg·L^-1, with detection limits (3S/N) in the range of 0.36-0.47 mg·kg^-1. Test for recovery was made by standard addition method, giving results in the range of 96.0%-101% and RSDs (n=6) all less than 3.0%.