In this application note, the key factors that affect the separation of lycopene, beta-carotene, lutein, and coenzyme Q10 by UPC2 were investigated.
UPC2 is suitable for the analysis of carotenoids and coenzyme Q10 due to the superior solubility of these non-polar compounds in the supercritical CO2 mobile phase. A fast separation of these bioactive compounds on an ACQUITY UPC2 HSS C18 SB Column was achieved in less than 6 minutes. Design of experiments was employed to investigate the individual and combined effects of ethanol percentage, temperature and pressure on the retention and separation factors. Retention factors were found to be strongly dependent on the amount of ethanol and pressure, followed by temperature whereas the three factors affected the separation. In this study, it was demonstrated that UPC2 enables a fast method development that can be used in routine analysis of carotenoids and coenzyme Q10.