Simultaneous Determination of Campanulin, Paeoniflorin and Hydroxysafflor Yellow A in Buyang Huanwu Decoction by HPLC

Objective To establish a HPLC method for the simultaneous determination of campanulin, paeoniflorin and hydroxysafflor yellow A in Buyang Huanwu Decoction. Methods A Kromasil C18(4.6 mm× 250 mm, 5 μ m) column was adopted. The mobile phase consisited of methanol and acetonitrile(26 ∶ 2)(A), and 0.7 % phosphoric acid solution(B), in gradient elution (0～ 10 min, 25 % → 35 % A; 10～ 25 min, 35 % → 40 % A).The flow rate was at 1.0 mL· min-1. The column temperature was 35 ℃ , and the detection wavelengths were set at 260nm for campanulin, 230 nm for paeoniflorin and 403 nm for hydroxysafflor yellow A. Results The linear ranges of campanulin, paeoniflorin and hydroxysafflor yellow A were 0.0539～ 1.078 μ g (r=0.999 5), 0.4160～ 8.320 μ g (r=0.999 8), and 0.0418～ 0.8352 μ g (r=0.999 5) respectively; the average recovery was 99.4 % (RSD=1.5 % ), 100.6 % (RSD=1.7 % ), and 101.0 % (RSD=1.9 % ) respectively. Conclusion The method is simple, feasible and reproducible, and can be used for the quality control of Buyang Huanwu Decoction.