ABSTRACT
OBJECTIVE To establish the fingerprints of Danggui buxue pills a nd the method for the content determination of three indicative constituents （ferulic acid ，calycosin 7-O-β-D-glucoside and astragaloside Ⅳ）. METHODS Fifteen batches of Danggui buxue pills from two manufacturers were analyzed by high performance liquid chromatography （HPLC）. The determination was performed on a Hypersil ODS 2 C18 column with mobile phase consisted of acetonitrile- 0.2％ formic acid （gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 25 ℃，and the sample size was 20 μL. UV detector [detection wavelengths were 250 nm （calycosin 7-O-β-D-glucoside），323 nm （ferulic acid ）] and evaporative light scattering detector （astragaloside Ⅳ）were selected as detectors. HPLC fingerprints of 15 batches of Danggui buxue pills were established with Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）. The chromatographic peaks were identified and assigned by comparing with the chromatogram of the reference substance and reference medicinal material ；the contents of ferulic acid ，calycosin 7-O-β-D-glucoside and astragaloside Ⅳ were also determined. RESULTS There were 33 common peaks in the fingerprints of 15 batches of samples with the similarities not lower than 0.893. Ferulic acid and calycosin 7-O-β-D-glucoside were identified as peak 13 and 15，respectively. Compared with the chromatogram of reference medicinal material，it could be found that peaks 1-3，7，8，10，12，13（ferulic acid ），17-19，27-29，32 and 33 belonged to Angelica sinensis，and peak 14，15（calycosin 7-O-β-D-glucoside），20-23，25 belonged to Astragalus membranaceus . The methodology of content determination met the requirements. The mean contents of ferulic acid ，calycosin 7-O-β-D-glucoside and astragaloside Ⅳ in 15 batches of samples were 0.050 1，0.402 6，0.913 4 mg/g. CONCLUSIONS In this study ，HPLC fingerprints of Danggui buxue pills and the method of HPLC quantitative analysis for three indicative constituents are established. Established methods are accurate，reliable and repeatable.