HPLC određivanje sfingozina i sfinganina u serumu kao potencijalnih biomarkera metaboličkog sindroma

master's thesis

• Title (english): HPLC determination of sphingosine and sphinganine in serum as potential biomarkers of metabolic syndrome
• Author: Andrea Križanac
• Mentor: Sunčica Beluhan (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biochemical Engineering, Industrial Microbiology and Malting and Brewing Technology); Zagreb
• Defense date and country: 2016-11-17, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

U ovom radu provedeno je određivanje koncentracije C18 sfingozina i C18-DL-erythrosfinganina ekstrahiranih iz uzoraka seruma zdravih ispitanika i ispitanika s dijagnosticiranim metaboličkim sindromom. Cilj rada je bio utvrditi mogu li navedene sfingoidne baze poslužiti kao specifični biomarkeri metaboličkog sindroma. Provedena je ekstrakcija ukupnih sfingolipida nakon čega su sfingoidne baze, C18 sfingozin i C18-DL-erythro-sfinganin, oslobođene iz kompleksnih sfingolipida metodom kisele... More hidrolize. Ekstrahirane sfingoidne baze analizirane su metodom tekućinske kromatagrafije visoke djelotvornosti (HPLC), a dobiveni rezultati su obrađeni statistički. HPLC analiza kod obje ispitivane skupine je pokazala sljedeće: od ukupnih sfingoidnih baza najzastupljenija baza je bila C18 sfingozin, zatim nepoznate OPA derivatizirane baze, dok je u odnosu na ukupne sfingoidne baze najmanje zastupljena bila baza C18-DL-erythro-sfinganin. Dobivene su vrijednosti koncentracije ukupnog C18 sfingozina kod zdravih ispitanika od 48,98 ± 12,80 nmol ml-1 (izraženo kao SV ± SD), a kod ispitanika s MetS 26,29 ± 9,63 nmol ml-1. Koncentracija ukupnog C18- DL-erythro-sfinganina kod zdravih ispitanika iznosila je 1,18 ± 0,54 nmol mL-1, dok je kod ispitanika s MetS iznosila 0,72 ± 0,40 nmol ml-1. Omjer C18-DL-erythro-sfinganina i C18 sfingozina kod zdravih ispitanika je iznosio 0,024 ± 0,009 , a kod ispitanika s MetS 0,027 ± 0,013. Pokazano je da ne postoji razlika u koncentracijama sfingoidnih baza između muškaraca i žena unutar svake ispitivane skupine. Utvrđena je statistički značajna razlika u koncentraciji C18 sfingozina (p<0,001) i C18-DLerythro-sfinganina (p=0,0032) izmeĎu dvije ispitivane skupine. Dobiveni rezultati ukazuju na promjenu metabolizma sfingolipida izazvanog metaboličkim poremećajem u organizmu. Less

Abstract (english)

In this work, quantitative changes of C18 sphinngosine and C18 sphingosine-DL-erythrosphinganine extracted from serum samples from healthy individuals and patients with diagnosed metabolic syndrome, have been determined. The aim of the study was to determine whether the sphingoid bases can serve as specific biomarkers of metabolic syndrome. Extraction was followed by acid hydrolysis to release the sphingoid base backbones, C18 sphingosine and C18-DL-erythrosphinganine, from the complex... More sphingolipids. Extracted sphingoid bases were determined by the method of high performance liquid chromatography (HPLC), and the results were statistically analysed. HPLC analysis in both groups showed the following: the most abundant base was C18 sphingosine followed by unknown OPA derivatized bases, while in relation to the total sphingoid base content, C18-DL-erythro-sphinganine was recovered as quantitatively minor fraction. The resulting value of concentration of total C18 sphingosine in healthy subjects was 48.98 ± 12.80 nmol mL-1 (expressed as SV ± SD) and in subjects with MetS 26.29 ± 9.63 nmol mL-1. The concentration of total C18-DL-erythro-sphinganine in healthy subjects was 1.18 ± 0.54 nmol mL-1 , whereas in subjects with MetS was 0.72 ± 0.40 nmol mL-1. The ratio of C18-DL-erythro-sphinganine and C18 sphingosine in healthy subjects was 0.024 ± 0.009, and in subjects with MetS 0.027 ± 0.013. Serum concentrations of total sphingoid bases showed no major variations between men and women within each group. There was statistically significant difference in the concentration of C18 sphingosine (p<0.001) and C18-DLerythro-sphinganine (p=0.0032) between two treatment groups. The results indicate a change of sphingolipid metabolism induced by metabolic disorder. Less

• Keywords: HPLC; metabolički sindrom; sfingolipidi; sfingozin; sfinganin
• Keywords (english): HPLC; metabolic syndrome; sphingolipids; sphingosine; sphinganine
• Language: croatian
• URN:NBN: urn:nbn:hr:159:258932
• Study programme: Title: Bioprocess Engineering; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva (magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2017-04-25 07:36:38