Novel stationary phases in HPLC for the analysis of biologically active compounds - determination of vitamine E

Bc. Jiří Plíšek, 2008 Application of technology new stationary phases in HPLC analysis of biologically active compounds - determination of vitamin E (Abstract) The aim of my graduation thesis was to find the acceptable stationary phase for pretreatment of biological material [blood plasma] and isolation of vitamin E (α-tocopherol) by SPE technique and subsequent determination by HPLC analysis by using the developed method. LLE was then altered by SPE. I used vitamin A (retinol) as standard for my own work, because there is a possibility that the simultaneous determination of both vitamins for the laboratory GMK FN HK will be an advantageous combination. The main emphasis was put on vitamin E. Sample preparation - optimized SPE procedure: Blood samples were drawn from the peripheral vein after twelve hours of overnight fast. The samples were then centrifuged (1600×g, 10 min, 4řC) and plasma was separated. Then 250 μl of plasma was added and deproteinized by absolute ethanol (625 μl, 10 min, 4řC). After centrifugation (1600×g, 15 min, 4řC), the supernatant was separated and applied on the SPE column. Before application of the sample, the SPE column was activated by 1 ml methanol and then washed by 1 ml of purified water. After application of the sample the precipitate was diluted in 625 μl of absolute..