Abstract
A high-performance liquid chromatographic electrochemical detection for the rapid and simultaneous determination of the vitamin A, D 3 and E is described. The separation is carried out by using a C 18 reversed-phase column and 0.1 M LiClO 4 in methanol-water (99:1, v/v) as the mobile phase. The compounds are eluted with good resolution in the above order within about 15 min and are determined by amperometric detection with a glassy carbon electrode at + 1050 mV ( vs. Ag/AgCl). The method gave reproducible results and the detection limits were of the order of 0.07, 4 and 0.2 ng of vitamin A, D 3 and E, respectively. The method was successfully applied to the determination of vitamin A, D 3 and E in liquid cow milk and milk powder samples. After saponification, fat-soluble vitamins were extracted with hexane and a methanolic solution of the dried extract was injected directly into the chromatographic system, avoiding the clean-up step that is necessary for vitamin D 3 when electrochemical detection is not used. Good recoveries were obtained.
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