Optimized HPLC extraction method of quercetin and berberine based on response surface analysis

Abstract

In order to establish a method for simultaneous determination and extraction of quercetin and berberine in soil, HPLC-PDA multi-wavelength method was used to detect the content of berberine and quercetin in soil solution. The detection wavelength was 210 nm and 347 nm. The column temperature was 30 °C, the mobile phase A was acetonitrile, the mobile phase B was 0.1% phosphoric acid aqueous solution, and the flow rate was 1.0 mL min⁻¹. Under the condition of isocratic elution, quercetin and berberine were completely separated within 20 min. The detection limit concentration of quercetin was 0.078 mg L⁻¹, and the detection limit of berberine was 0.019 mg L⁻¹. Both of them reached the trace level, and the recovery rate was between 97.2% and 107.4%. The response surface method was used to optimize the ultrasonic extraction method. The three main factors of extraction concentration, extraction temperature and solid–liquid ratio were optimized to obtain the highest extraction efficiency. The optimum extraction efficiency was as follows: 1 g soil sample was extracted with 80% ethanol aqueous solution, ultrasonic time was 10 min, ultrasonic temperature was 44 °C, and solid–liquid ratio was 1 : 17 g mL⁻¹. The extracted quercetin and berberine concentrations were close to the predicted values of response surface optimization. The method of extracting and determining berberine and quercetin from soil established in this experiment is simple, fast, low cost and high safety. The feedback of the results also further verifies the feasibility in practical production and application, and provides reference value for further research and analysis of different allelochemicals in soil.