Determination of Six Components in Shensu Xuanfei Wan by HPLC with Wavelength Switching Combined Gradient Elution Method

OBJECTIVE To develop an HPLC with wavelength switching combined gradient elution method for simultaneous determination of caffeic acid, rosmarinic acid, 3'-hydroxy puerarin, puerarin, 3'-methoxypuerarin and daidzin in Shensu Xuanfei Wan. METHODS ZOBAX SB-C₁₈ column (4.6 mm×250 mm,5 μm) was applied with methanol-acetonitrile (1:1) (A) and 0.1% glacial acetic acid solution(B) as mobile phase in gradient mode. The flow rate was 1.0 mL·min^-1. Caffeic acid and rosmarinic acid were detected at 320 nm, 3'-hydroxy puerarin, puerarin, 3'-methoxypuerarin and daidzin were detected at 250 nm. Injection volume was 20 μL. RESULTS The concentration and peak area of caffeic acid, rosmarinic acid, 3'-hydroxy puerarin, puerarin, 3'-methoxypuerarin and daidzin were at 2.32~46.40 μg·mL^-1(r=0.999 8), 3.06~61.20 μg·mL^-1 (r=0.999 5), 4.45~89.00 μg·mL^-1(r=0.999 9), 14.48~289.60 μg·mL^-1(r=0.999 9), 4.86~97.20 μg·mL^-1(r=0.999 7), 3.69~73.80 μg·mL^-1 (r=0.999 6), respectively, and showed a good linear relationship. The average recoveries and the corresponding RSD were 99.0%(1.6%), 97.5% (0.6%), 99.3%(0.9%), 98.6%(1.3%), 97.6%(1.3%), 97.0%(0.9%), respectively. CONCLUSION The method is accurate, quick, simple. It could be reliable for the quality control of Shensu Xuanfei Wan.