Quality assessment of Zaoren-an-shen granule by High-performance liquid chromatography (HPLC) fingerprinting and quantitative analysis

Abstract

To control the quality of Zaoren-anshen granule (ZRG), a simple, reliable and reproducible method of High-Performance Liquid Chromatography equipped with Photodiode Array Detector (HPLC-DAD) was developed for fingerprint analysis and quantitative determination of three major classes of constituent’s namely phenolic acids, flavonoids and lignanoids. In the fingerprint analysis, nineteen peaks were selected as characteristic peaks and the chemical characteristics of three herbs in ZRG were presented in the HPLC chromatographic file. In the quantitative analysis, one flavonoid spinosin, four phenolic acids including danshensu, protocatechuic acid, protocatechuic aldehyde and salvianolic B and four lignanoids including schisandrin, gomisin A, deoxyschizandrin and gomisin N were successfully separated on an UltimateTM XB-C18 column (250 × 4.6 mm I.D., 5.0 μm) at 35°C. The mobile phase was acetonitrile-water (containing 0.03% phosphoric acid) employing a gradient elution at a flow rate of 1.0 mL/ min, and the detection wavelength was set at 280 nm. Regression equations revealed good linear relationship between the peak areas of the analytes and their concentrations (r > 0.9991) and the recoveries were in the range of 95.5~103.7%. The results indicated that the developed assay method could be considered as a suitable quality control method for ZRG. Key words: Zaoren-anshen granule, high-performance liquid chromatography, quality control, fingerprint.