Status of methodology for the determination of fat-soluble vitamins in foods, dietary supplements, and vitamin premixes.

Fat-soluble vitamins (FSVs) include vitamin A, carotenoids, vitamins D, E, and K. New legislation is being introduced in many countries to reinforce regulatory compliance of declared concentrations of vitamins and other micronutrients in food products and dietary supplements. The levels of FSVs are likely to be more closely scrutinized due to their potential health risks associated with overdosing, in particular of vitamin D. However, a proviso of stricter regulatory compliance is that analytical methods must be fit-for-purpose, providing adequate accuracy and precision. Official methods have been published by organizations such as AOAC INTERNATIONAL, European Committee for Standardization, International Dairy Federation, U.S. Pharmacopeia, and International Organization for Standardization. The methods available for foods, dietary supplements, and vitamin premixes are evaluated in this review. In general, these methods show adequate precision for regulatory compliance; however, the field of application has not often been evaluated for a sufficiently large range of food matrixes. Gaps have been noted in the range of published official procedures, particularly for carotenoids and vitamin premixes. The potential of some recent developments in sample preparation and chromatographic techniques were evaluated to provide improved procedures for FSV analysis the future.

Simultaneous quantification of vitamins A, D3 and E in fortified infant formulae by liquid chromatography-mass spectrometry.

A novel method for the simultaneous quantification of Vitamins A, D3 and E in fortified infant formulae has been developed using isocratic normal-phase liquid chromatography with positive atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS). Food products were saponified and the vitamins were extracted by solid-phase extraction (SPE) on a Chromabond XTR cartridge. Quantification of Vitamins D3 and E were performed with Vitamin D2 and 5,7-dimethyltocol (DMT) as internal standards (IS), respectively while no IS was used for Vitamin A. Detection of the vitamins was made in the selected ion monitoring (SIM) mode. MS calibration curves were linear between 0.15 and 12 mg/l for Vitamin A, 5-400 microg/l for Vitamin D3 and 0.25-20 mg/l for Vitamin E with regression coefficient r2 > 0.996 and the limits of detection were below 1.4 ng. The repeatability (CV) obtained on a reference dietetic infant formula was 2.3% for Vitamin A, 2.6% for Vitamin E and 5.9% for Vitamin D3. The between-day variations (CV) over 6 days were in the ranges of 2.4-6.9% for the three vitamins. The mean recoveries from a reference infant formula spiked with all three vitamins ranged from 96 to 105% with a relative standard error less than 9%. The applicability of the method was demonstrated by analyzing a set of infant formula and infant cereals; similar results were obtained with the LC-MS method and reference HPLC methods.