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The Development of HPLC-FLD Method for the Determination of the Bile Acids in the Bile of Mice
Kacerovská, Veronika ; Nesměrák, Karel (advisor) ; Bosáková, Zuzana (referee)
Aim of the master thesis was the development and the optimization of the HPLC method with fluorescence detection for the determination of the bile acids in the bile of mice. On the basis of literature, the HPLC-FLD method has been proposed and optimized for the quantification of eight bile acids (cholic acid, glycocholic acid, deoxycholic acid, glycodeoxycholic acid, chenodeoxycholic acid, glycochenodeoxycholic acid, α-muricholic acid, and λ-muricholic acid) in the biological material. 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-p-toluenesulfonate is suitable derivatization reagent for labeling of the bile acids. The calibration dependences, limits of detection and quantification (in the range of 0.88-1.78 μmol dm -3 ) for all bile acids are measured. In the biological sample five bile acids have been successfully identified and then determined (cholic acid, glycocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, and α-muricholic acid). Key words: bile acids, HPLC, fluorescence detector, derivatization reaction, bile
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