Carotenoid Analysis by HPLC-DAD

HPLC-DAD separation was performed using a Shimadzu LC20 AT HPLC system (Shimadzu Corporation, Kyoto, Japan) with an SPDM20A diode array detector and a YMC C30 reversed-phase column (250 mm length, 4.6 mm inner diameter and 5 μm particle size). The experimental conditions for the separation and identification by HPLC-DAD were the same as described in a previous study [62 (link)]. Quantification of carotenoids was performed using external calibration with standards of β-carotene, lutein, β-cryptoxanthin, and zeaxanthin purchased from Extrasynthese (Lyon, France) in the range of 1–100 μg/mL. The HPLC-DAD analysis was performed three times for the tested sample and data are expressed in mg/100 g F.W (fresh weight) and presented as the mean ± SD of these three measurements.