Determination of Chlorogenic Acid, Forsythin, Rhein, Emodin and Chrysophanol in Lianhuan Qingwen Capsules by HPLC

OBJECTIVE To develop an HPLC method for determination of chlorogenic acid, forsythin, rhein, emodin and chrysophanol in Lianhuan Qingwen capsules. METHODS The sample was separated on an Agilent C₁₈ column(4.6 mm× 250 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.1% acetic acid in gradient elution(0 min→10min→20min→30min→45min→53min, acetonitrile 10%→10%→20%→30%→68%→75%) at the flow rate of 1.0 mL?min^-1；at 0-15 min, the detection wavelength was 327 nm, at 15-35 min, the detection wavelength was 205 nm, at 35-43 min, the detection wavelength was 230 nm, at 43-48 min, the detection wavelength was 221 nm, at 48-55 min, the detection wavelength was 224 nm, and the column temperature was 32 ℃. RESULTS The results showed that all the components were well separated in 55 min by HPLC. Each main component chromatographic peak separation resolution was >2.0, the linear ranges and correlation coefficients were 0.444 5-7.112 μg(r=0.999 6), 0.167 8-2.684 μg(r=0.999 9), 0.031 0-0.496 3 μg(r=0.999 5), 0.031 5-0.504 μg(r=0.999 7), 0.065 5-1.048 μg(r=0.999 9), respectively. The sums of average recovery(n=9) were 102.2%(RSD=0.5%), 101.0% (RSD=0.8%), 98.32%(RSD=0.5%), 95.83%(RSD=0.6%), 102.65%(RSD=0.3%). CONCLUSION The method is convenient, accurate, and can be used for quantitative determination of the preparation.