Simultaneous Chiral Separation of Flavanone, Naringenin, and Hesperetin Enantiomers by RP-UHPLC-DAD.

A rapid and effective RP-UHPLC-DAD method for enantioseparation of three flavanones, i.e., flavanone, naringenin, and hesperetin, was developed and validated. Chromatographic separation of the analytes was performed using a Chiralpak AD-3R analytical column under reverse phase conditions with methanol as the mobile phase. The method was validated in the concentration range of 0.2 to 50 µg/mL for enantiomers of flavanone and 0.5 to 50 µg/mL for enantiomers of naringenin and hesperetin. The limits of quantification were between 0.03 to 0.5 µg/mL. Intraday and interday precision were below 14% and accuracy varied from 0.04 to 8.17%.