Simultaneous Determination of Pinoresinol Diglucoside, Amygdalin and Cinnamylaldehyde in Bushen Huoxue Granule

OBJECTIVE To establish an HPLC method for determination of pinoresinol diglucoside, amygdalin, and cinnamylaldehyde in Bushen Huoxue granule. METHODS Agilent Zorbax Extend-C₁₈(4.6 mm×150 mm, 5 μm) was adopted; the mobile phase was acetonitrile-water with gradient elution at a flow rate of 1.0 mL?min^-1, and the detection wavelength was 210 nm(0-5 min) for amygdalin, 227 nm(5?10 min) for pinoresinol diglucoside, 290 nm(10-14 min) for cinnamylaldehyde. The methodological study included sample pre-treatment process and method validation. RESULTS The components of pinoresinol diglucoside, amygdalin, and cinnamylaldehyde were well separated, the method had a good linearity in the range of 0.426 0-2.88 6 μg?mL^-1 7.455-86.60 μg?mL^-1and 0.440 6?13.22 μg?mL^-1(r≥0.999); the average recoveries were 98.3% (RSD=1.6%), 101.4%(RSD=1.1%), 100.0%(RSD=1.2%), respectively, and the RSDs of intra-day and inter-day assays were all ≤1.4%. CONCLUSION This method was simple, sensitive and reproducible and can be used for the quality control of Bushen Huoxue granule and its related preparation.