Comparison of conventional and supercritical isolation methods from bee pollen and their effects on the concentrations of bioactive compounds

The present work represents a profound investigation of material pretreatment, comparison of different isolation methods (conventional and supercritical), and characterization of phenolic compounds and amino acids from bee pollen. The determination of amino acids was carried out after developing a novel hydrophilic interaction chromatography combined with quadrupole time-of-flight tandem mass spectrometric method (HILIC-QTOF-MS/MS) with limits of detection over the range 1 mg/kg −5 mg/kg and recoveries over the range 87.6 % - 97.1 %. The optimal conditions for the isolation of amino acids are in the range of temperatures between 25°C to 55°C and at pressures between 300 bar and 350 bar. It can be concluded that the extraction temperature and the pressure affect antioxidant activity. The favored conditions are lower temperatures and higher pressures. • Bee pollen offers bioactive compounds with antioxidant and anticancer properties. • UAE and SCF showed the best recovery of target compounds optimizing TP and vit. C. • Optimal amino acid extraction: 25–55°C and 300–350 bar, favoring high efficiency. • ABTS method revealed the highest antioxidant values, dominated by hydrophilic phenols. • Extracts inhibited melanoma cells by up to 25 %, sparing healthy melanocytes.