反应详情

Sunflower oil (7.5 g) and retinyl palmitate (2.5 g) were mixed and water (0.03 g) and immobilised Lipase D (Rhizopus oryzae (Amano) immobilised on Accurel® EP100 macroporous polypropylene (Acordis) 0.1 g) or Candida rugosa AY (Amano) on Accurel® EP100 (Acordis) (CR; 0.1 g) added. The mixtures were placed in a shaking water bath at 55° C. for 20 or 23 hours. The immobilised lipase was then removed by simple filtration to directly yield the product, a solution of retinol esters in triglyceride oil. The composition of the retinyl esters was separated from the triglycerides by thin layer chromatography (0.5 mm silica G plates, Analtech Ltd.) using toluene as eluting solvent and visualised by spraying with 1% 2,7-dichlorofluorescein in ethanol. The ester band was scraped off and FAMEs were produced using 3 mL sodium methoxide and 1 mL toluene at 80° C. for 20 minutes followed by 5 minutes at 80° C. with boron trifluoride (2 mL volumes of reagent). The fatty acid content was then analysed by Fatty acid methyl ester Gas Chromatography (FAME-GC). GC conditions: Column—30 m/0.53 mm/0.5 um restek famewax, Helium carrier gas 15 kpa, flame ionisation detection, 260° C. PTV injection (Run: 3 min @ 260° C., 80° C. hold 1 min, +20/min to 180° C., +2/min to 220° C., +1/min to 230° C., +4 min hold at 230° C.).

1. customThe mixtures were placed in a shaking water bath at 55° C. for 20 or 23 hours
2. customThe immobilised lipase was then removed by simple filtration
3. customto directly yield the product
4. customThe composition of the retinyl esters was separated from the triglycerides by thin layer chromatography (0.5 mm silica G plates, Analtech Ltd.)
5. washas eluting solvent
6. customFAMEs were produced
7. customat 80° C.
8. customfor 20 minutes
9. customPTV injection (Run: 3 min @ 260° C., 80° C. hold 1 min, +20/min to 180° C., +2/min to 220° C., +1/min to 230° C., +4 min hold at 230° C.)