Separation of Retinol Acetate, Vitamin a Palmitate, Α- and Γ- Tocopherol by Reverse-Phase High Performance Liquid Chromatography and Determination of These Vitamins in Van Lake Fish (Chalcalburnus Tarichi)

Abstract

The resolution properties of several commercial columns for vitamin A and E standards by reverse phase chromatography methods have been examined. Hypersil MOS column with methanol-water (96:4) and acetonitrile-water (90:10) mobile phase, Shimadzu-ODS column with methanol-water (96:4) mobile phase have acceptable resoluion in separation of vitamin A esters. Shimadzu-ODS column/96 % methanol and Shimadzu-C8/90% acetonitrile have the acceptable resolution ratio for vitamin E isomers. Methanol extracts of heart muscle and gill in Van Lake fish (Chalcalburnus tarichi) have been examined for vitamin A and E by high performance liquid chromatography. Avarage retinol acetate, vitamin A palmitate, alpha tocopherol, gamma tocopherol in heart muscle of Van Lake fish, have been found as 162±23 ng/g, 18±2.2 ng/g, 8.5±1.0 μg/g, 505.7±94 ng/g, respectively. The vitamin levels in gills have been determined as retinol acetate 231.5±38 ng/g, vitamin A palmitate 23.8±3.5 ng/g, alpha tocopherol 12.4±0.77 μg/g, gamma tocopherol 350.1±97 ng/g. A significant differences have been found among the retinol acetate-vitamin A patmitate and alpha-gamma tocopherol values in heart muscles and gills (P<0.05). A negative correlation for retinol acetate between heart muscle and gill (r:-.86), vitamin A palmitate between heart muscle and gill (r:-.73) and between retinol acetate-alpla tocopherol in heart muscles (r:-.71) have been observed. A positiv correlation between retinol acetate and alpha tocopherol in gills (r:+.52) has been found. No correlation has seen between heart muscles and gills for gamma tocopherol values.