HPLC analysis of carotenoids in particular carrot (Daucus Carota L.) cultivars

UDC 635.132; 633.432; 547.912

HPLC analysis of carotenoids in particular carrot (Daucus Carota L.) cultivars

Zelenkova E. N., Yegorova Z. E., Shabunya P. S., Fatykhava S. A.

Optimal conditions for analysis of carotenoids in carrot by HPLC, including sample preparation, were selected. Chromatograph Agilent 1200 with diode array detector was used. The sample components were separated on column ZORBAX Eclipse Plus C18 (3.0×100 mm; 1.8 μm) at temperature 22 deg C. The temperature in the autosampler was 10 deg C, injection volume – 2 μl. The isocratic solvent system consisting of acetonitrile – methanol – ethyl acetate (73:20:7, v/v/v) was chosen as the mobile phase. The flow rate of eluent was 0.4 ml/min. It is shown that 50 ml of acetone was needed for extraction of carotenoids from the 5 g of homogenized carrot samples (10-multiple excess). Separating capacity of the solvents (acetone, the mixture acetonitrile – methanol – ethyl acetate, dichloromethane) and their compatibility with the mobile phase was investigated. It was established experimentally that the acetone is the best solvent to recover dry residue of pigments. Developed method was used for the study of carotenoids in eleven carrot cultivars, zoned in Belarus. The main carotenoids of carrots were distributed as follows: α-carotene – 30-40 %, β-carotene – 55-68 %, lutein – 1-6% of carotenoid total amount. Cultivars Vitaminnaya-6 and Dordon were characterized by significant amount of β-carotene (90–100 µg/g) and α-carotene (53–61 µg/g). The largest quantity of lutein (4.3–6.5 µg/g) was contained in cultivars Dordon, Morelia, Nerac and Niland. The highest content of carotenoids (~155 µg/g) was observed in cultivars Dordon and Vitaminnaya-6. Carrot cultivars grown in Belarus were characterized by average contents of β-carotene, but the carotenoid total amount was high. Described HPLC method can be applied for the determination of the main carotenoids of carrot.

Keywords: carotenoids, analysis, HPLC method, carrot, cultivar

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