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Voltammetric detection of 27-hydroxycholesterol on boron-doped diamond electrode
Michnová, Kristýna ; Schwarzová, Karolina (advisor) ; Nesměrák, Karel (referee)
The aim of this bachelor thesis was to study the voltammetric behavior of 27-hydroxycholesterol using boron-doped diamond electrode. The influence of several factors on the current response of 27-hydroxycholesterol was investigated: the effect of the supporting electrolyte, concentration of perchloric acid, polarization rate, water content, and pH effect. The study was conducted using cyclic voltammetry in a perchloric acid or sodium perchlorate in acetonitrile. 27-hydroxycholesterol provided a current response in the anodic region with a potential around +1450 mV vs. Ag/0.01 mol l-1 AgNO3 in 1 mol l-1 NaClO4 in acetonitrile in the presence of 0.1 mol l−1 perchloric acid in acetonitrile (water content 0.43 %) and 0.1 mol l−1 sodium perchlorate in acetonitrile. Furthermore, concentration dependence was measured using differential pulse voltammetry resulting in a limit of detection (LOD) of 2.71 μmol l−1 and a limit of quantitation (LOQ) of 9.04 μmol l−1 . It was demonstrated that the direct current voltammetry method does not achieve comparable limit values. Key words 27-hydroxycholesterol, boron-doped diamond electrode, oxidation, oxysterols, voltammetry
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Voltammetry and liquid chromatography of cholesterol, its precursors, and oxidation products
Kubešová, Jana ; Schwarzová, Karolina (advisor) ; Baluchová, Simona (referee)
Electrochemical detection of sterols is difficult. This thesis aims to study the voltammetric behaviour of selected oxysterols, i.e. 25-hydroxycholesterol and 7-ketocholesterol on the boron-doped diamond electrode. Further, an HPLC method with spectrophotometric and electrochemical detection was developed for separation of selected sterols. The study was performed using cyclic and differential pulse voltammetry. 0.1 mol l−1 perchloric acid in acetonitrile (water content 0.55 %) was chosen as the most suitable electrolyte. 25- hydroxycholesterol provides anodic signal at the potential of +1500 mV and 7-ketocholesterol at +1900 mV in that media. Electrochemical reduction of 7-ketocholesterol was also studied. However, the observed cathodic signal isn't reproducible. The stability of oxysterols in the presence of perchloric acid was confirmed by UV-VIS spectroscopy. Calibration dependence of oxysterols was measured using differential pulse voltammetry and resulted in the limit of detection of 0.82 µmol l−1 for 25-hydroxycholesterol and 0.66 µmol l−1 for 7-ketocholesterol. In the second part of the thesis, an HPLC method was developed for the separation of a sterol mixture on a reversed-phase C18 column using a mobile phase containing 50 mmol l−1 sodium perchlorate in acetonitrile with 0% or 6% water...
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Voltammetric determination of sulfathiazole on boron doped diamond electrode
Zich, Mikuláš ; Fischer, Jan (advisor) ; Schwarzová, Karolina (referee)
This bachelor thesis is dedicated to study of the electrochemically active substance sulfathiazole. Sulfathiazole is an antibacterial agent added to animal feed. As a result, it contaminates soil, water bodies and food. A boron-doped diamond electrode was chosen as the working electrode for voltammetric determination. First, the optimal pH for the study of sulfathiazole in the range of 2 to 12 by the technique differential pulse voltammetry was determined and the optimal pretreatment of the electrode before measurements. Subsequently, the calibration dependence was measured. Sulfathiazole was successfully dissolved in water at a concentration of c = 1·10-3 mol l-1 and Britton-Robinson buffer was used as the electrolyte. As the optimal pH was determined pH 6,0. At this pH, a relative standard deviation of sr = 5% was achieved. Standart deviation of 5 % has been achieved when the working electrode was polished before each measurement. Under these conditions, was calibration dependence measured in concentration range from 0,1 to 100 μmol l-1 . Linear dynamic range was calculated between 0,1 - 20 μmol l-1 . Limit of quantitation was found to be 97 nmol l-1 .
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Voltammetric detection of 7α-hydroxycholesterol on boron doped diamond electrode
Serbanová, Lucie ; Schwarzová, Karolina (advisor) ; Nesměrák, Karel (referee)
This bachelor's thesis is dedicated to the electrochemical determination of 7α-hydroxycholesterol (7α-OHC) using boron-doped diamond electrode (BDDE). The work includes the optimization of experimental conditions and the measurement of the concentration dependence of 7α-OHC. The influence of various factors, such as water content in the solution and scan rate, on the electrochemical response of 7α-OHC was investigated. Various electrochemical methods, including cyclic voltammetry and differential pulse voltammetry, were used in the experiments. The electrochemical behavior of 7α-OHC in a solution of chloric acid and acetonitrile were analyzed. Important factors such as electrode passivation and response stability were carefully evaluated. The measurement of concentration dependence of 7α-OHC exhibited a linear response within the concentration range from 2∙10−5 mol l−1 to 1∙10−4 mol l−1 . Limits of detection (LOD) and quantification (LOQ) of 1,33 µmol l−1 and 4,43 µmol l−1 were determined. Comparison with other methods revealed that the electrochemical determination of 7α-OHC on BDDE achieves comparable sensitivity. The results of this work provide crucial insights into the electrochemical behavior of 7α-OHC, highlighting the advantages of BDDE for sensitive and reliable determination of...
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Modification of boron doped diamond electrode by electrochemical oxidation of p-aminobenzoic acid
Švárová, Eliška ; Fischer, Jan (advisor) ; Dejmková, Hana (referee)
This bachelor's thesis deals with the modification of a boron-doped diamond electrode (BDDE) by the electrochemical oxidation of p-aminobenzoic acid. Subsequently, the stability of the modified electrode was investigated by a study of various parameters. The modification of the electrode was performed using cyclic voltammetry (CV) in LiClO4 electrolyte. The success of the modification and its subsequent stability was investigated by the CV method in electrolyte KCl in the presence of the redox system [Fe(CN)6]4-/3- . Stability was tested in terms of ultrasonic cleaning in isopropanol and anodic cleaning in 0.5 mol l-1 H2SO4. It was found that the modification of the surface cannot be completely removed by ultrasonic cleaning in isopropanol, in contrast, pulses of period 2 seconds with a high positive potential are enough to completely remove the modification. Optimal conditions for surface stability were determined. The ideal number of cycles for surface modification was set at 5 cycles. The use of different electrolytes during modification (KClO4 and LiClO4) determined that in the case of modification in the KClO4 electrolyte, it is possible to measure in the range of the potential window from -400 to +1600 mV and in the case of LiClO4 in the range of -400 to +1500 mV without breaking the modified...
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Electrochemical study of the DNA desorption from the surface of carbon electrodes
Průcha, Jakub ; Dejmková, Hana (advisor) ; Schwarzová, Karolina (referee)
This bachelor's thesis deals with the possibility of DNA desorption from the surface of carbon electrodes. Measurements were performed using linear scan voltammetry in a three- electrode configuration, with single-stranded DNA (ssDNA) of 31 nucleotides as the sample. The working electrodes were a boron-doped diamond electrode and an electrode made of highly oriented pyrolytic graphite. The efficiency of desorption was assessed by comparing the heights of oxidation signals of adsorbed nucleic bases, which were measured before and after the application of the desorption procedure. The first approach involved desorption through mixing or rinsing in solutions with different properties, specifically in organic solvents: methanol, acetone, toluene, and DMSO; in acids and bases: HCl, HNO3, NaOH; and subsequently in surfactants and detergents: Triton X-100, TWEEN 80, CTAB, and Tergazyme. Among the tested substances, a 0.1 mol·l-1 solution of NaOH and a 1 % solution of Tergazyme proved to be effective. Subsequently, the time and temperature dependence of the rinsing strength of these two solutions on ssDNA desorption were examined. From the conducted experiments, it was determined that time and temperature of the solution do not increase the efficiency of the desorption by NaOH solution. For rinsing with a...
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Testing of the electrochemical properties of flow-through diamond detector
Pecák, Jakub ; Dejmková, Hana (advisor) ; Vyskočil, Vlastimil (referee)
In the presented bachelor thesis, the functionality of a flow-through multitubular boron- doped diamond electrode (mtBDDE) was verified on two selected neurotransmitters, DA and NE, using the HPLC technique with isocratic elution. An aqueous solution of Britton-Robinson (B-R) buffer was chosen as the mobile phase and a Kinetex EVO C18 reverse column (150 x 4.6 mm) as the stationary phase. The flow rate of the mobile phase was set at 1 ml/min. For electrochemical detection, mtBDDE was used and UV detection at 280 nm was applied as an assistant to the main detector. The electrochemical response of the DA and NE solution (cΘ = 1.10-4 mol/dm3 ) was investigated in the form of hydrodynamic voltammograms over a potential range from 0.1 V to 1.3 V in different pH ranges of the mobile phase (2.5; 4.0; 6.0; 8.0 and 10.0). It was found that the potential wave shifts to lower potentials with increasing pH of the mobile phase, while the response of individual analytes does not change significantly with pH. Under optimal conditions, i.e., mobile phase B-R buffer at pH = 6 and a potential loaded on the mtBDDE of 0.6 V, the repeatability and reproducibility of the measurement results were verified. The electrode gave repeatable results (RSD < 4 %) comparable to the UV detector used (RSD ≤ 4 %) over the series of...
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A new approach to the electroanalysis of primary bile acids and related steroids
Klouda, Jan ; Schwarzová, Karolina (advisor) ; Skopalová, Jana (referee) ; Vyskočil, Vlastimil (referee)
In this doctoral thesis, a novel method for the determination of primary bile acids cholic acid and chenodeoxycholic acid is presented. Bile acids play various vital roles in the mammalian body. Moreover, their determination is extremely helpful in liver and biliary disease diagnosis and management. These saturated organic compounds lack strong chromophores and fluorophores in their structure, and thus are usually hard to detect in spectroscopy. For this reason, either instrumentally advanced but expensive methods, such as mass spectrometry, or less reliable enzymatic methods are commonly employed in bile acids quantitation. Hence, the demand for simple and reliable methods for their determination is strong. Bile acids are also known to be virtually inert for direct electrochemical oxidation. Herein, a simple method for their chemical activation for electrochemical oxidation on bare electrode materials was developed, optimized and applied to cholic acid and chenodeoxycholic acid determination. The activation is based on a dehydration reaction of a primary bile acid with 0.1 mol L-1 HClO4 in acetonitrile (water content 0.55%) that introduces double bond(s) into the originally fully saturated steroid core. This naturally increases the electron density in the structure, and thus allows electrochemical...
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Development of voltammetric methods for detection of phytosterols on carbon-based electrodes
Zelenský, Michal ; Schwarzová, Karolina (advisor) ; Fischer, Jan (referee)
The aim of this diploma thesis was to develop an electrochemical method for three phytosterols, namely stigmasterol β-sitosterol and ergosterol using voltammetry techniques on glassy carbon electrode and boron doped diamond electrode. The work builds on previous results, where it was shown that the electrochemical oxidation of phytosterols requires perchloric acid in acetonitrile. When this acid is used, a dehydration reaction occurs. Calibration dependences were measured here for all phytosterols in an environment with a water content of 0.43%. The stability of the signal with increasing distilled water content was further investigated. Then the calibration dependences in the increased water content were measured again. If we want to use HPLC-type separation techniques, we must first find out how the phytosterols will behave in this environment, and therefore the signal stability on the buffer content was investigated. The buffers used were phosphate and acetate buffer. Subsequently, the concentration dependence was measured in an environment with a higher content of phosphate buffer. Furthermore, the possibility of detecting two phytosterols at once was investigated. Key words: dehydration, glassy carbon electrode, boron doped diamond electrode, oxidation, phytosterols, voltammetry
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Development of electroanalytical methods for detection of bile acids possessing 7α hydroxyl group
Jelšíková, Kristýna ; Schwarzová, Karolina (advisor) ; Dejmková, Hana (referee)
This master's thesis contains a study of electrochemical processes of selected bile acids possessing 7 hydroxyl group (cholic, chenodeoxycholic and −muricholic). The measurements were performed on boron−doped diamond electrode in the non-aqueous medium of acetonitrile and perchloric acid (water content 0.55 %) by cyclic voltammetry. It is known that the electrochemical activity of 7 bile acids is increased by a dehydration reaction between perchloric acid and the 7 bile acid. The subject of the study was the stability of the voltammetric response of chemically activated bile acids in the region of negative potentials. It was found that the presence of oxygen in the measured solution is an important factor for obtaining the cathodic signal of 7 bile acids. It probably performs a regenerative function; the product of the electrochemical reduction is re-oxidized in its presence, which leads to an increase in the voltammetric response. At the same time, it is important that the direction of the scan in cyclic voltammetry first proceeds to positive values. A potential of +2.0 V (vs. Ag/AgNO3 in acetonitrile) must be reached for the HO● radicals to be formed. It is these radicals that presumably lead to the formation of the product(s) of bile acids electrochemical oxidation that can be subsequently...
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