ABSTRACT
Objective To establish a method for simultaneous determination of berberine, magnolol and emodin in the blend powder of Cortex Phellodendri Chinensis, Cortex Magnoliae Officinalis and Radix et Rhizoma Rhei, so as to provide evidence for preparation technique and quality control of simplified recipe of Ruyijinhuangsan. Methods High-performance liquid chromatography (HPLC) was used for simultaneous determination of berberine, magnolol and emodin at the following conditions: the column was ACE.5C18-AR, mobile phase was A: triethylamine, B: methanol. The flow rate was 1.0 mL/min, the temperature of column was 30°C, the detection wavelengths were set at 294 nm and 345 nm, and the injection volume was 10 μL. Using the yield ratio of three constituents as the assessment indices, we optimized the extraction technology by single factor test and orthogonal test L9 (34). Results The calibration curves were linear within the range of 0.292-292 mg/L for berberine (r2=0.999 8), 0.292-292 mg/L for magnolol (r2=0.999 7), and 0.256-256 mg/L for emodin (r2=0.999 5), with the average recovery rates being 98.26% (RSD 2.53%, n=6), 105.17%(RSD 2.06%, n=6), and 100.39% (RSD 2.60%, n=6), respectively. The results of precisions were (n=6) 0.23%, 0.17%, 0.18%, respectively. The sample solutions were stable within 12 h. The single factor experiment analyzed ultrasonic duration and times, concentration of alcohol, the amount of solvent and the concentration of HCl. The results showed that, with the increase of ultrasonic duration, times and the alcohol concentration, the yield ratio of the three consitituents was increased.Conclusion The optimum options for extracting is as follows: ultrasonic extraction 3 times by 95% alcohol containing 0.1% HCl, with the volume at 35 mL, 30 min each time.