ObjectiveTo establish a quantitative analysis of multi-components by single-marker(QAMS)for simultaneously determining uridine, guanosine, lucidenic acid C and ganoderic acid A in Compound Anoectochilus roxburghii(Wall.)Lindl (A.roxburghii)oral liquid, and validate its feasibility for quality evaluation.MethodsThe contents of uridine, guanosine, lucidenic acid C and ganoderic acid A in oral liquid were determined by high-performance liquid chromatography(HPLC), and the separation was performed on ZOBAX SB-Aq C 18 column(4.6 mm×250 mm, 5 μm).The mobile phase comprised acetonitrile-ammonium acetate(13 mmol/L ammonium acetate, pH=4)with the gradient elution and at a flow rate of 1.0 mL/min.The wavelength for detection and column temperature were set at 254 nm and 30 ℃, respectively.The injection volume was 20 μL.Using guanosine as the internal reference substance the relative correction factors(RCFs)for uridine, lucidenic acid C and ganoderic acid A was determined by HPLC-UV with good reproducibility.To verify the accuracy and feasibility of the QAMS, the contents of four active ingredients were also determined by the external standard method(ESM).ResultsThe linear range of uridine, guanosine, lucidenic acid C and ganoderic acid A were 2-40 μg/mL( r²=0.999 9), 3-60 μg/mL(r²=0.999 9), 1-20 μg/mL(r²=0.999 9)and 1-160 μg/mL( r²=0.999 9), respectively.RCFs of uridine, lucidenic acid C and ganoderic acid A with reference to guanosine were 1.02, 0.34, 0.40, respectively.No significant differences between the quantitative results of QAMS and ESM were observed(S=0.999 9).The average recoveries(n=3)were 94.5%, 104.0%, 86.6% and 105.0%, respectively.ConclusionThe established QAMS cab be used for the determination of four active ingredients in compound A.roxburghii oral liquid.