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This bachelor's thesis deals with the development of a method for the separation of carotenoid pigments, especially astaxanthin, β-carotene and lutein, produced by the microscopic green alga Tetracystis pulchra. The work compares published methods for the separation of carotenoid pigments and attempts to optimize a method suitable for the analysis of pigment extracts from Tetracystis pulchra cells. Optimized conditions for HPLC-DAD separation of carotenoid pigments were as follows: mobile phase A - ACN/MeOH/0,05M Tris-HCl buffer (pH = 8,0): 94/2/4, mobile phase B - MeOH/EtOAc: 68/32 under gradient elution, flow rate 0,25 ml min˗1 , column temperature 28 řC. The column used was Kinetex C18 (100 × 2,1 mm, 2,6 µm), injection volume was 20 µl and UV detection was performed at 470 nm. Total analysis time was 23 minutes. Key words: HPLC, astaxanthin, β-carotene, lutein |
