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Analysis of arginine and its methylated derivatives in clinical samples using LC-MS/MS
Prokop, Pavel ; Boušová, Iva (advisor) ; Raisová Stuchlíková, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Pavel Prokop, DiS. Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of diploma thesis: Analysis of arginine and its methylated derivatives in clinical samples using LC-MS/MS Asymmetric dimethylarginine (ADMA) and its enantiomer, symmetric dimethylarginine (SDMA), are naturally occurring amino acids that are generated intracellularly from L-arginine. ADMA is an endogenous competitive inhibitor of nitric oxide synthase. Increased levels of ADMA and SDMA are associated with endothelial dysfunction, hypertension, cardiovascular and renal diseases. Both compounds are accumulating in chronic kidney disease (CKD) patients. The aim of this diploma thesis was to develop and validate a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for the determination of L-arginine, ADMA and SDMA in human plasma. Subsequently, LC-MS/MS method was compared with enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography with fluorescence detection (HPLC-FL) method. Chromatographic separation was performed on Dionex UltiMate 3000 UHPLC liquid chromatograph, followed by tandem mass spectrometric detection on amaZon SL mass spectrometer. L-Arginine-13 C6, ADMA-d6 and...
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