Vývoj a optimalizace nové chromatografické metody pro stanovení retinolu v moči

• Main Author: Mikanová, Zuzana
• Other Authors: Kujovská Krčmová, Lenka
• Format: Dissertation
• Language: Czech
• Published: 2018
• Online Access: http://www.nusl.cz/ntk/nusl-380637

Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Zuzana Mikanová Supervisor: assoc. prof. RNDr. Lenka Kujovská Krčmová, Ph.D. Title of the Diploma Thesis: Development and optimization of new chromatographic method for determination of retinol in human urine This diploma thesis is based on the method development for determination of retinol and creatinine using HPLC. The aim of this project was to find optimal HPLC-MS/MS conditions for clinical research. Experiment was carried out using UHPLC set Nexera with mass spectrometer LCMS- 8030, (Shimadzu, Japan). Three stationary phases were tested. The chromatographic separation was achieved using a Kinetex 2.6 µm PFP 100A 100x4.6 mm (Phenomenex, USA). The used mobile phase consisted of acetonitrile (with addition of formic acid 0.005 M) and ammonium acetate (pH 6.69 ) in the ratio 78:22 (v/v). The temperature was maintained at 25řC, flow rate was set at 0.5 ml/min and 3 µl of sample was injected. After optimization of separation conditions, the method was applied to urine samples, and simple sample preparation procedures were tested In these days is the work on method finishing and the new method will be used for determination of renal damage in clinical research and practice. The inclusion of...