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1 Wiadomości Chemiczne

1 2001

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Wykorzystanie HPLC do oznaczania witamin rozpuszczalnych w tłuszczach

Wieliński S., Olszanowski A.

Wiadomości Chemiczne

2001

[Z] 55, 11-12

1075-1102

In this review, selected chemical techniques such as TLC, UV-VIS, GC, HPLC and microbiological procedures (published since 1960) used for simultaneous determination of fat-soluble vitamins in different matrices have been discussed. Particular attention has been given to HPLC methods, not only in terms of chromatographic conditions, but also in comparison with other separation techniques. HPLC methods have been shown to have clear advantages over existing chemical or microbiological assays in terms of sensitivity, specificity and sample throughput for the analysis of these vitamins. In order to assist the analyst in the selection of a particular procedure, certain chromatographic conditions have been listed for this purpose. However, as currently, no specific HPLC procedures can be recommended, it was decided to develop new methods for these vitamins. Within the past 30 years, HPLC has become the predominant method for separation and quantification of fat-soluble vitamins. The two major, distinct methods of HPLC separation are referred to as normal-phase chromatography and reverse-phase chromatography. To generalise, reverse-phase HPLC is frequently preferred for analysis of biological samples as the columns are more easily purged of any contaminants and sample separation is usually less sensitive to slight changes in mobile-phase composition. A summary of HPLC systems used for fat-soluble vitamins analysis is provided in Table 1 [60-106] and 2 [107-152] in format: compounds determined, matrix, sample preparation and clean up, column, mobile phase, detection, time of analysis, reference. Finally, this paper describes a procedure for the simultaneous determination of vitamin A (retinol acetate, palmitate), vitamin D3 (cholecalciferol), vitamin E (alpha-tocopherol acetate) and alphacalcidol in capsules from a single sample extract using reverse phase HPLC and column backflushing techniques [162-165]. The procedure eliminates saponification, lengthy extractions and sample workup. It is specific to the compounds of interest, shows very good internal precision and is free of interference compared to current published methods.