Simultaneous Determination of Five Bioactive Compounds in Polygonum cuspidatum by HPLC
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Abstract:
Five bioactive compounds namely polydatin, resveratrol, emodin-8-O-β-D-glucoside, emodin and physcion in Polygonum cuspidatum were determined by HPLC. An Agilent Zorbax SB C18 column (250 mm×4.6 mm i.d., 5 μm) was used. The mobile phase consisted of acetonitrile (A) and water (B) with linear gradient program of 15.0~50.0% (A) in 0~40 min and 50.0~90.0% (A) in 40~60 min. The column temperature, detection wavelength and flow rate were set at 40 ℃, 290 nm and 1.0 mL/min respectively. The calibration curve of each analyte was constructed under six concentrations. The correlation coefficients were all more than 0.999, and the RSD of tR ranged from 0.14% to 1.67%, while RSD of the peak area was from 0.96% to 2.96%. The spike recovery of the five analytes was 96.8~104.7%. The extraction condition was optimized, and ultrasonic extraction with 60% ethanol for 30 min was chosen. The method was successfully applied for analysis of ten Polygonum cuspidatum samples. The results showed that polydatin and emodin-8-O-β-D-glucoside were the two most dominant constituents in Polygonum cuspidatum with contents ranges of 9.80~20.42 mg/g and 5.56~24.45 mg/g, respectively. The method has good repeatability and high precision, and can be used for simultaneous analysis of the five constitutes in Polygonum cuspidatum.
