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Determination of L-tryptophan in food supplements in high performace liquid chromatography
Pešková, Pavlína ; Kalíková, Květa (advisor) ; Kubíčková, Anna (referee)
This bachelor's thesis deals with optimalization of chromatographic conditions for determination of tryptophan by HPLC method with UV detection. The amount of tryptophan has been determined in two food supplements. To find suitable conditions for the determination of tryptophan, the influence of the composition of the mobile phase on analyte retention and peak symmetry was tested in two different chromatographic modes - reversed-phase mode using the Atlantis dC18 column and mixed-mode using the Atlantis PREMIER BEH C18 AX column. Specifically, the effect of the type and amount of organic modifier (acetonitrile/methanol), the effect of the pH of the ammonium acetate buffer (4,7;5,9;7,0) and its concentration (5mM/10mM) in the mobile phase was tested. Based on the obtained data, the optimal chromatographic conditions were selected (in terms of analysis time and peak symmetry). In the optimized conditions selected validation parameters such as repeatability, reproducibility, limits of detection and quantification were determined. The optimized conditions were as follows: mixed-mode Atlantis PREMIER BEH C18 AX column, mobile phase acetonitrile/5mM ammonium acetate, pH 7,0 10/90 (v/v), separation temperature 25 řC, flow rate 1 ml/min and UV detection at 254 nm. Under these conditions, the tryptophan...
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