|
|
Utilization of vibrational spectroscopy in study on interactions between humic substances and organic ions.
Stehlíková, Kristína ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
The main objective of this bachelor thesis was to validate the potential of the utilization of vibrational spectroscopy in the study on interactions between humic substances and organic ions. Unmodified (HA) and methylated humic acids (mHA) were chosen as model humic substances and organic ions were represented by methylene blue, rhodamine 6G and Septonex. Firstly, a suitable infrared spectroscopy technique was chosen. The results of optimization measurements showed that spectra with the best resolution can be obtained using the DRIFTS technique. The next part of the thesis discusses the effect of methylation in the structure of humic acids. Significant differences were recorded in mHA compared to the unmodified humic acids. Finally, the spectra of humic acid – organic ion complexes were compared to the spectra of prepared blank samples. It was noted that the spectra of complexes are not only a simple sum of the pure individual substances’ spectra, but also shifts and changes in the intensity of different absorption bands were recorded there. It suggests that non-covalent interactions played an important role in the complexes. Infrared spectroscopy was proved to be a useful method for investigating these interactions.
|
|
|
Raman spectroscopy as a tool for analysis of biotechnologically relevant microorganisms
Záhorská, Linda ; Enev, Vojtěch (referee) ; Mgr.Ota Samek, Ph.D. (advisor)
The diploma thesis deals with the study of biotechnologically significant microorganisms, using the Raman spectroscopy. Content of the theoretical part is brief characteristic of Raman spectroscopy as a method, its use in practice and also use as a tool for monitoring of biotechnologically processes. Thesis was further focus on the biotechnologically significant microorganism Aureobasidium pullulans, its use in biotechnology and also for over-produced substances and in particular poly-L-maleic acid and pullulan. The content of the experimental part was study of selected strains A. pullulans, specifically stains as DSMZ, CCM F148 and CCM 8182, using Raman spectroscopy on the various types of culture media. Subject of practical part research was too production of extracellular polymers, acid poly-L-apple and pullulan, by selected strains A. pullulans. Objective of my thesis was described and determinate, spectra of individual strains as well as extracellular products, mainly pullulan, and then choose suitable production medium and optimal production strain A. pullulans. During experimental work was found, that optimal production strain was DSMZ strain culture on the mineral medium with the addition of yeast autolysate, which was optimal medium type. The content of the pullulan produced was for gravimetric determination, 6,3g/L, which also confirmed the results of the HPLC method. It was experimentally found, that Raman spectroscopy isn´t suitable method for quantification of extracellular products, but is appropriate and was used for PCA analysis of individual strains.
|
|
|
Transport of pharmaceuticals in soil systems
Janebová, Denisa ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
Drugs have a negative effect on nature. If they do not degrade in wastewater treatment plants, they can contaminate lakes, rivers and even drinking water. However, the soil contains soil organic matter, which has chelating properties and can prevent the migration of the drug. The aim of this bachelor thesis was to map the transport of the selected drug in the soil column and try to determine its diffusivity. Sulfapyridine was chosen for transport. Sulfapyridine belongs to a group of sulfonamide antibiotics that are often detected in the environment. It is mainly used in veterinary medicine to treat various infections. The partial goal was to describe the basic properties of the soil depending on the depth of the soil layer. The drug concentration profile in the soil column was determined by UVVIS spectrometry in soil extracts. The following reagents were used for leaching: distilled water, as a pure solvent without the presence of other substances, tap water as a model reagent replacing the soil solution and magnesium chloride solution, which is used for soil extracts of freely moving and ionically bound substances in the soil.
|
|
|
Fractionation and molecular organization of humic acids
Chytilová, Aneta ; Enev, Vojtěch (referee) ; Klučáková, Martina (advisor)
Humic acids are part of the natural organic matter occurring all around us. The aim of this thesis is to study the molecular organization, conformation of humic acids in aqueous solutions, that always raise a number of questions. For a long time, the scientists all over the world argue, if humic acid are polymers, micelles or supramolecules. Over time, thanks to new technologies, their opinions are moving away from a polymer model and tend rather to supramolecular arrangement of humic acids. Studying humic complex systems is not easy, because they are polydisperse and heterogeneous, which significantly complicates any characterization. Moreover, its molecular organization is affected by many factors such as e.g. pH, ionic strength and etc. For the study of the conformation of humic acids, concentration series of IHSS (International humic substances asociation) Leonardite humic acids stamdards in four different mediums were prepared: 0,1 M NaOH, humic acids in water with pH modified to 12 (basic medium), 0,1 M NaOH + 0,1 M HCl, 0,1 M NaCl (neutral environment). Furthermore it has been performed the fractionation of humic acids for the purpose of simplifying the complicated structure. Prepared concentration series were characterized with several analytical methods such as ultraviolet and visible spectroscopy, dynamic light scattering, electrophoretic light scattering, microrheology, gel permeation chromatography, potentiometric pH determination and direct conductometry. Diploma thesis is built on the previous bachelor thesis in which different sample of humic acids was studied. The measured results indicate that the studied systems show supramolecular behavior and in some cases are subject to aggregation into larger units (micelles).
|
|
|
Study of bulk and hydration water in hydrogel systems suitable for medical applications
Řihák, Marek ; Sovová, Šárka (referee) ; Enev, Vojtěch (advisor)
This bachelor thesis focuses on a preparation of hydrogel systems based on hyaluronan and Septonex and on a design and optimalization of quantitative analysis of hyaluronan and Septonex in a supernatant after gelation. This work compares hydrogels formed from hyaluronan of two different molecular weights. The calibration curves of hyaluronan and Septonex were used to evaluate the experimental data. The thermogravimetric analysis and Fourier-transform infrared spectroscopy were used for the characterization of properties of the samples. The utilisation of the mentioned techniques to study molecular water subpopulations was discussed according to the obtained results.
|
|
|
Distribution of the Organic Matter in a Sample of Biochar
Novotný, Maximilián ; Enev, Vojtěch (referee) ; Kalina, Michal (advisor)
This bachelor’s thesis is focused on characterization of the content and distribution of organic matter in a sample of biochar. For these purposes, the organic matter from a biochar sample was isolated using two procedures, specifically sequential chemical fractionation, and alkali extraction. Prepared samples were characterized by elemental analysis and thermogravimetry. Analysis results showed that sequential chemical fractionation brings higher yields than alkali extraction, moreover, it provides more detailed image of organic matter in biochar thanks to obtained separated fractions according to their linkage to the original biochar matrix. Secondly, studied biochar has a great potential as soil supplement, it contains almost 70 % of organic carbon, whilst isolated amount, which is available represent only small amount, the rest is strongly bound. On the other hand, the study showed that this specific biochar has rigid structure, which brings benefits in the context of using biochar in agriculture in long-term consideration.
|
|
|
Bioactive substances for face cream: Determination of stability of face cream incorporated with goat's milk
Dostálová, Tereza ; Sovová, Šárka (referee) ; Enev, Vojtěch (advisor)
The aim of this bachelor thesis is a stability trial of a skin cream incorporated with goat milk lyophilizates prepared in usual laboratory conditions. Firstly, the goat milk was denatured, to be further analysed via FTIR, elemental analysis and thermogravimetry. To compare the denaturation effect, measurements with native goat milk sample were performed as well. Next, an emulsion was prepared. First stability tests were performed using analytical centrifuge and the cream preparation procedure and component ratio was optimized. Stability trials were carried out at specified time intervals. Rheological properties – especially yield stress values and structure strength, determined by linear viscoelastic region and complex modulus – thermal endurance measured via differential scanning calorimetry and visual validation via optical microscopy and sample observation were tested and performed. Skin cream stability of sample stored at laboratory temperature was demonstrated, showing moderate sings of structure destabilisation. Product suitability for dry skin was proved by sensory analysis. The skin feeling was pleasant, and the cream moisturized the surface area, that it was applied to, according to evaluators. Visually, the emulsion appeared to maintain its properties during the experiment time range, it did not undergo a complete degradation of separation into two original phases, nor it was degraded, for example by mold.
|
|
|
Sequential fractionation of organic matter of humic acid isolated from Leonardite
Krist, Tomáš ; Doskočil, Leoš (referee) ; Enev, Vojtěch (advisor)
The aim of the diploma thesis was to optimize the sequential fractionation method of organic matter to be used for physico-chemical characterization of extracted fractions. Humic acid isolated from oxidized brown coal of Leonardite was used as a source matrix of organic matter. An eluotropic series was assembled and sequential fractionation was performed by extraction on a Soxhlet apparatus. The original humic acid and fractions were characterized by elemental analysis (EA) and thermogravimetric analysis (TGA), followed by Fourier transform infrared spectrometry (FTIR), molecular absorption spectrometry (UV/VIS), fluorescence spectrometry and potentiometric titration. Atomic ratios were determined from the results of the elemental analysis. From the measured UV/Vis and fluorescence excitation and emission spectra, the absorption coefficients, resp. fluorescence coefficients. Used fractionation method proved to be a suitable method for studying HA structure. A total of 62 wt. % of initial materiál was extracted, indiivdual fraction amounted from 0.36–30.92 wt. %. From the results of the structural analysis, it is clear that with increasing polarity of the organic solvent, fractions with long aliphatic chains were first isolated and their aromaticity graddualy increased. Non-polar organic solvents were suitable for the extraction of lipid-like coumpounds, while the most polar organic fractions were rich in polar groups and their structual parameters were close to the original humic acid. The fraction extracted with acetonitrile was the most unique fraction. This fraction was rich on nitrogen and amine groups and was similar to protein-like structures. In the last two fractions, extracted with alcohols, a significant bathochromic shift typical of fluorophore type V was observed. Among other things, they were also characterized by a higher content of plant carbohydrate residues.
|
|
|
Utilization of physico-chemical and spectroscopical methods in study on microbial extremofiles
Dobešová, Kateřina ; Enev, Vojtěch (referee) ; Sedláček, Petr (advisor)
The aim of this bachelor thesis is the determination od polyhydroxyalkanoates in the cells of extremophille bacterie, preformed by several methods. The chosen bacterium was Schlegelella thermodepolymerans DSM 15 344 Thermogravimetric analysis, infrared spectroscopy with Fourier transformation, and Raman spectroscopy were used in this thesis. The TGA method was used for the determination of extracellular and intracellular water in samples. The FTIR method was used in order to observe the amount od PHA, but aslo to observe the physical properties of the polymer in the sample, specifically to determine either amorphous or crystalline featuers of this polymer. Raman spectroscopy was used in oder to confirm the presence of PHA in the samples. All results e´were compared eith the results accomplished by gas chromatografy.
|
|
|
Program-controlled freeze drying for the preparation of the delivery system of native hyaluronan and hydrophobic molecules
Waczulíková, Kristína ; Sedláček, Petr (referee) ; Enev, Vojtěch (advisor)
The amphiphilic nature of the hyaluronan biopolymer facilitates interactions between its hydrophobic portions of the chain and the hydrophobic ligands, making it one of the promising compounds that can be used as a suitable carrier for drug delivery. Therefore, the aim of this diploma work was by means of a program-controlled freeze-drying to prepare a system based on native hyaluronan, which would be capable of transporting hydrophobic drugs. Perylene, pyrene, prodan and 1,6-diphenyl-1,3,5-hexatriene fluorescence probes were used as model compounds to simulate hydrophobic drug-hyaluronan interactions. Freeze-drying efficiency was studied using infrared spectrometry and scanning electron microscopy. ATR-FTIR spectrometry confirmed for all probes but pyrene the hydrophobic interactions between the hyaluronan and probe molecules based on the presence of the C=C bond valence vibration absorption band in the aromatic probe rings. From the IR spectra, it was not possible to confirm with certainty the presence of residual tert-butyl alcohol in the samples. SEM photographs revealed that increasing the pressure during drying process had a positive effect on the quality of the lyophilized sample in the second series of samples. Interactions of individual fluorescent probes with hyaluronan were studied using steady-state and time-resolved fluorescence spectrometry. Positive results have been obtained, indicating the presence of fluorescent probes interacting with the hydrophobic regions of the hyaluronan chains. The results of this work can contribute to the development of carrier systems in the field of targeted drug distribution.
|
guest ::
