One step assay of the four plant ingredients in Yangxue Anshen syrup by HPLC method

Objective  To establish a HPLC method for simultaneous determination of quercitrin, luteoloside, rutin and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in Yangxue Anshen syrup. Methods  Waters symmetry C18 column (250 mm×4.6 mm, 5 μm) was used with 0.1% acetic acid (A) and methanol (B) as the mobile phase. Gradient elution was performed at a flow rate of 1.0 ml/min, 0-15 min, 95%-90%A; 15-35 min, 90%-70%A; 35-55 min, 70%-60%A; 55-85 min, 60%-50%A; 85-95 min, 10%A. The detection wavelengths were 256 nm and 320 nm. Column temperature was 30 ℃ and the injection volume was 10 μl. Results  Quercitrin, luteoloside, rutin and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside showed good linear relationship within the range of 10-300, 5.0-150.0, 5.0-150.0, 20.0-600.0 µg/ml(r≥0.9989), respectively. The average recovery was (96.75±1.41)%, (99.61±1.01)%, (97.18±1.96)% and(99.12±0.97)% (n=6), respectively. Conclusion  The established method is simple, accurate and stable, which can be used for the simultaneous determination of 4 components in Yangxue Anshen syrup.