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Study of lipid biosynthesis during hypoxia using chromatography and mass spectrometry
Mušutová, Martina ; Cvačka, Josef (advisor) ; Čabala, Radomír (referee)
5 Abstract The topic of this diploma thesis is the study of lipide biosynthesis in a hypoxic environment with the use of chromatography and mass spectrography. The first part of the thesis explains the theoretical basis for the research stipulated in the study of OSA and its connection to Type 2 diabetes mellitus. I propose the theory that hypoxia leads to the heightened biosynthesis of fatty acids by way of the reductive citric acid cycle. The research of this reductive citric acid cycle was done by means of cultivated cells with added labeled [5-13 C] glutamine and its absorption into the fatty acids has been observed with the help of GC-MS. The lipides have been extracted from the cell samples, from which the fraction of triglyceride has been isolated with the help of thin layer chromatography. Furthermore, the transesterification to fatty acid methyl esters (FAME) has been performed. The collected data showed a 91 % increase in FAME 16:0 and a 102 % increase in FAME 16:1 in a hypoxic environment compared to the control group. Furthermore, it has been found that samples in a hypoxic environment contained 5.9 % more [13 C1] FAME 16:0, 12 % more [13 C1] FAME 16:1 and almost 3 % more [13 C2] FAME 16:1 compared to the control group. Keywords Lipide biosynthesis, Type 2 diabetes mellitus, FAME, GC-MS,...
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Study of lipid biosynthesis during hypoxia using chromatography and mass spectrometry
Mušutová, Martina ; Cvačka, Josef (advisor) ; Čabala, Radomír (referee)
5 Abstract The topic of this diploma thesis is the study of lipide biosynthesis in a hypoxic environment with the use of chromatography and mass spectrography. The first part of the thesis explains the theoretical basis for the research stipulated in the study of OSA and its connection to Type 2 diabetes mellitus. I propose the theory that hypoxia leads to the heightened biosynthesis of fatty acids by way of the reductive citric acid cycle. The research of this reductive citric acid cycle was done by means of cultivated cells with added labeled [5-13 C] glutamine and its absorption into the fatty acids has been observed with the help of GC-MS. The lipides have been extracted from the cell samples, from which the fraction of triglyceride has been isolated with the help of thin layer chromatography. Furthermore, the transesterification to fatty acid methyl esters (FAME) has been performed. The collected data showed a 91 % increase in FAME 16:0 and a 102 % increase in FAME 16:1 in a hypoxic environment compared to the control group. Furthermore, it has been found that samples in a hypoxic environment contained 5.9 % more [13 C1] FAME 16:0, 12 % more [13 C1] FAME 16:1 and almost 3 % more [13 C2] FAME 16:1 compared to the control group. Keywords Lipide biosynthesis, Type 2 diabetes mellitus, FAME, GC-MS,...
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Determination of Lead using Sequential Injection Analysis with Spectrophotometric Detection
Mušutová, Martina ; Hraníček, Jakub (advisor) ; Rychlovský, Petr (referee)
The main objective of the thesis was the determination of lead concentration in model aqueous solutions of 4-(2-Pyridylazo)-resorcinol complexating reaction with lead cation in 2-Amino-2-hydroxymethyl-propane-1,3-diol hydrochloride buffer (TRIS buffer). The UV/VIS molecular spectrometry in static arrangement was chosen with subsequent application of sequential injection analysis in flow arrangement. Calibration curve was measured and both methods were assessed. Detection threshold was 0,026 mg dm-3 in case of static arrangement and 0,70 mg dm-3 in case of flow arrangement. The interference analysis was performed, with metallic ions (Cu2+ , Zn2+ , Sn4+ ) being the most significant interferents, which also create a chromatic complex with 4-(2-Pyridylazo)-resorcinol. The interference effects were weak and almost not present for barium and potassium. Thesis concludes with determination of lead concentration in model specimen using flow arrangement. The determination by calibration curve was insufficient due to interfering ions and thus it had to be determined by standard addition method. Keywords Lead, 4-(2-pyridylazo)-resorcinol, TRIS buffer, flow sequential injection analysis, UV/VIS molecular spectrometry, interference effects
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