The purpose of this study was to analyze the effect of the type of extraction solution (water, different concentrations of ethanol), temperature and time on the polyphenol content and antioxidant properties of red clover extracts and the effect of the addition of selected extracts on the antioxidant properties of enriched blackcurrant beverages. In both the extractions carried out under different conditions and in the enriched beverages, the content of selected polyphenols was determined by HPLC. This study confirmed the significant effect of the alcohol content of the extract, extraction time and temperature on the antioxidant properties of clover extracts. Ethanolic extracts had better antioxidant properties than aqueous extracts. The addition of ethanol extracts had a significant effect on the antioxidant properties of the fortified beverages. Increasing the temperature, time or ethanol content in the extracts mostly resulted in an increase in the total polyphenol content in the obtained extracts. Based on the analysis of the response surface, it was found that for the DPPH radical, the best activity was obtained by extraction for 20 min with a solution of approximately 65% at low temperatures. In the case of the ABTS radical, the best antiradical activity was obtained after extraction for 60 min at 80 °C with a solution of approximately 50% ethanol. It was also found that the use of a solution of approximately 60% ethanol after extraction for 60 min at 80 °C would provide an extract with high antiradical activity against both radicals.
Keywords: HPLC; antioxidant activity; extraction conditions; optimization; polyphenols; red clover.