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Development of HPLC method for evaluation of tryptophan metabolites
Pšenčíková, Barbora ; Kastner, Petr (advisor) ; Kučera, Radim (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Barbora Pšenčíková Supervisor: PharmDr. Petr Kastner, Ph.D. Title of thesis: Development of HPLC method for evaluation of tryptophan metabolites The essential amino acid tryptophan is a precursor of several important bioactive compounds. In recent years it was found out that tryptophan metabolites are involved in the pathogenesis of many diseases, including neurodegenerative and psychiatric disorders, autoimmune diseases, or cancer. Therefore, the analytical methods for simultaneous determination of tryptophan and its metabolites are highly required. The purpose of this diploma thesis was to develop an optimal HPLC method for determination of tryptophan and its metabolites (kynurenine, kynurenic acid, serotonin and 5-hydroxyindole-3-acetic acid) and to consider all the possible options of determination of quinolinic acid and melatonin. The chromatographic separation was carried out by Supelco Ascentis Express F5 column, 2.7 μm particle size, 15 cm × 3 mm, using spectrophotometric and fluorescence detection. Parameters of detection were for kynurenine UV detection 369 nm, 254 nm, fluorescence detection Ex/Em: 369/475; for kynurenic acid UV detection 244 nm; for...
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Development of HPLC method for evaluation of tryptophan metabolites
Pšenčíková, Barbora ; Kastner, Petr (advisor) ; Kučera, Radim (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Pharmaceutical Analysis Candidate: Barbora Pšenčíková Supervisor: PharmDr. Petr Kastner, Ph.D. Title of thesis: Development of HPLC method for evaluation of tryptophan metabolites The essential amino acid tryptophan is a precursor of several important bioactive compounds. In recent years it was found out that tryptophan metabolites are involved in the pathogenesis of many diseases, including neurodegenerative and psychiatric disorders, autoimmune diseases, or cancer. Therefore, the analytical methods for simultaneous determination of tryptophan and its metabolites are highly required. The purpose of this diploma thesis was to develop an optimal HPLC method for determination of tryptophan and its metabolites (kynurenine, kynurenic acid, serotonin and 5-hydroxyindole-3-acetic acid) and to consider all the possible options of determination of quinolinic acid and melatonin. The chromatographic separation was carried out by Supelco Ascentis Express F5 column, 2.7 μm particle size, 15 cm × 3 mm, using spectrophotometric and fluorescence detection. Parameters of detection were for kynurenine UV detection 369 nm, 254 nm, fluorescence detection Ex/Em: 369/475; for kynurenic acid UV detection 244 nm; for...
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