HPLC Analysis of Vitamins

HPLC Analysis of Vitaminsin Tablets using HPLCAbstractFat-soluble vitamins, such as vitamins E, D, and A, and water-soluble vitamins, such as vitaminsC, B6, B2, B1, and B12, have been analyzed.Vitamins are biologically active compounds that act as controlling agents for an organism’snormal health and growth. The level of vitamins in food may be as low as a few micrograms per100 g. Vitamins often are accompanied by an excess of compounds with similar chemicalproperties. Thus not only quantification but also identification is mandatory for the detection ofvitamins in food. Vitamins generally are labile compounds that should not exposed to hightemperatures, light, or oxygen. HPLC separates and detects these compounds at roomtemperature and blocks oxygen and light.1 Through the use of spectral information, UV-visiblediode-array detection yields qualitative as well as quantitative data. Another highly sensitive andselective HPLC method for detecting vitamins is electrochemical detection.0 2 4 6 8 10 12Norm050010001500Citric acidStandardB1 B6Pantothenic acidB12BiotinRiboflavinFolic acid, diCaVitamintabletSaccharinTime [min]Column 100 ˘ 4 mm Hypersil BDS, 3 μmMobile phaseA= water with pH = 2.1 (H2SO4) = 99%B = ACN + 10% A = 1 %Gradientat 3.5 min 1% B; at 11 min 25% Bat 19 min 90% BPost time 6 minFlow rate 0.5 ml/minColumn compartment 30 ºCInjection vol 2–5 μlDetectorUV-DAD detection wavelength 220/30 nm,reference wavelength 400/100 nmSample preparationFiltrationConditionsAngelikaGratzfeld-HuesgenFoodFigure 1Analysis of water-soluble vitamins in a vitamin tabletAgilent TechnologiesInnovating the HP WaySample preparationDifferent food matrices require different extraction procedures.1 Forsimple matrices, such as vitamin tablets, water-soluble vitamins canbe extracted with water in an ultrasonic bath after homogenization ofthe food sample.Chromatographic conditionsThe HPLC method presented here was used to analysis vitamins in avitamin drink.HPLC method performanceLimit of detection