Development of a green chromatographic method for determination of fat-soluble vitamins in food and pharmaceutical supplement

Abstract

A green chromatographic analytical method for determination of fat-soluble vitamins (A, E, D 3 and K 1) in food and pharmaceutical supplement samples is proposed. The method is based on the modification of a C18 column with a 3.00% (w/v) sodium dodecyl sulphate (SDS) aqueous solution at pH 7 (0.02 mol L −1 phosphate buffer solution) and in the usage of the same surfactant solution as mobile phase with the presence of 15.0% (v/v) butyl alcohol as an organic solvent modifier. After the separation process, the vitamins are detected at 230 nm (K 1, D 3 and E), 280 nm (A, E, D 3 and K 1) and 300 nm (K 1, D 3 and E). The chromatographic procedure yielded precise results (better than 5%) and is able to run one sample in 25 min, consuming 1.5 g of SDS, 90 mg of phosphate and 7.5 mL of butyl alcohol. When the flow rate of the mobile phase is 2 mL min −1 the retention times are 4.0, 9.6, 13.0 and 22.7 min for D 3, A, E and K 1 vitamins, respectively; and all peak resolutions are higher than 2. The analytical curves present the following linear equations: area = 6290 + 34852 (vitamin A), R 2 = 0.9998; area = 4092 + 36333 (vitamin E), R 2 = 0.9997; area = −794 + 30382 (vitamin D 3) R 2 = 0.9998 and area = −7175 + 82621 (vitamin K 1), R 2 = 0.9996. The limits of detection and quantification for vitamins A, E, D 3 and K 1 were estimated for a test pharmaceutical vitamin supplement sample as 0.81, 1.12, 0.91 and 0.83 mg L −1 and 2.43, 3.36, 2.73 and 2.49, respectively. When the proposed method was applied to food and pharmaceutical sample analysis, precise results were obtained (R.S.D. < 5% and n = 3) and in agreement with those obtained by using the classical chromatographic method that uses methanol and acetonitrile as mobile phase. Here, the traditional usage of toxic organic solvent as mobile phase is avoided, which permits to classify the present method as green.