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Zavedení a optimalizace metodiky pro stanovení lipofilních a hydrofilních vitamínů v biologických vzorcích
Kalábová, Vendula
The main scope of this diploma thesis are the methods used for building vitamins, especially priorizing chromatographic methods. You can also find the characteristics of vitamins dissolvable in fat and water, their physiological meaning, daily use, presence in foodstuffs and methods, which are suitable for their analysis. The experimental part describes the sample preparation, their vitamin extraction methods, the requierements for chromatographic assessions, methods of qualitative and quantitative evaluation of chromatographs with the focus on the calibration curve method. The work also puts an emphasis on establishing the limit of detection and the limit of quatification, and determining the lipophilic vitamins in matrices rich of these vitamins using the method of HPLC.
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The use of HPLC technique in the analysis of phenolic compounds in plant material of fruit trees
Adamcová, Anežka ; Šatínský, Dalibor (advisor) ; Sklenářová, Hana (referee) ; Vávra, Radek (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Anežka Adamcová Supervisor: Prof. RNDr. Dalibor Šatínský, Ph.D. Title of the Dissertation Thesis: The use of HPLC technique in the analysis of phenolic compounds in plant material of fruit trees Although fruits are well studied and known for their high content of antioxidants and positive health benefits, the main aim of the presented dissertation thesis consists in the characterization of biological active compounds in individual parts of fruit tree material (leaves, bark, buds, flowers, and wood chips). The theoretical part is focused on fruit trees and the characterization of phenolic compounds. Their properties and the effects based on current scientific studies are shown. A large part of thesis is dealing with the chromatographic methods for the separation of phenolic substances including the current trends. The extraction process is also included. The practical part deals with the validation and development of screening HPLC-DAD method for the analysis of monitored phenolic compounds (phloridzin, phloretin, chlorogenic acid, quercitrin, rutin, arbutin, 1,5-dicafeoylquinic acid, and 3,5-dicafeoylquinic acid) in individual parts of fruit trees. The main objective of this thesis is to...
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Assassement of the Active Ingredient Delta-9 THC in Biological Marerial and Hemp Drugs
VONDRÁŠKOVÁ, Eliška
This bachelor's thesis deals with the matter of abusing hemp drugs and its products in the past as well as at present day. The thesis focuses on possible contributions to well-being of using hemp but also health risks, which come with the abuse of hemp and hemp drugs. The aim of this thesis is to partly chart the data about THC abuse from three South Bohemian hospitals. The hospitals are: the hospital of České Budějovice, the hospital of Písek and the hospital of Strakonice. The data involve the number of tests done in hospital facilities in order to confirm or to confute the presence of THC in biological material, most commonly in urine. The thesis describes preanalytical and analytical phase of sample treating where mistakes must be avoided. The bachelor's thesis is than occupied with the principles of the assessment of THC in biological material in both qualitative and quantitative ways of assessment of active ingredients. It contains findings gained during practice in a form of personal assistance and data from which is then made the conclusion of this thesis. The aim of this bachelor's thesis also involves charting of hemp abuse in a part of South Bohemian Region during past few years. This bachelor's thesis can serve as a base for those, who are interested in the discussed topic. Another contribution can be an extension of general knowledge amongst students and public, considering the curative benefits and the effect of hemp abuse at the same time. In the conclusion the bachelor's thesis concerns with how many tests were positive and how many were negative during the years charted in a certain hospital but also compares the hospitals mutually. It involves the development of hemp drug abuse through time in each region.
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Příprava mutantního serpinu z klíštěte \kur{Ixodes ricinus}
EDEROVÁ, Monika
Point mutation altering arginin for tryptophan amino acid residue in P1 site of tick salivary serpin Iripin-1 was created using specific primers. Recombinant protein with this mutation in nucleotide sequence was then expressed in chemically competent Escherichia coli cells, extracted from them and purified by affinity and size-exclusion chromatography. To see the impact of the mutation on inhibitory function of Iripin-1, its ability to bind trypsin and form covalent complexes was evaluated.
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Determination of glycohemoglobin by HPLC method
KUNÍKOVÁ, Martina
This bachelor thesis deals with the determination of the glycated hemoglobin by the HPLC method at two analyzers D-10 from the company Bio-Rad. The sample values for the internal control quality were measured and evaluated for two devices in the period from November 2019 to March 2020 in this bachelor thesis. The regular verification of the method was realized for both analyzers. The SEKK samples were measured in the planned survey of external quality assesment SEKK Compensation of diabetes KD 1/20. The regular comparation of both analyzers was performed by the measurement of laboratory samples once in the month. Overall 50 samples were measured in this period. The results of this bachelor thesis are part of a monitoring of the laboratory determination quality control and they are the base for the improving of the work quality in the laboratory and the base for the increase of the effectiveness of the laboratory's quality management system. Key words
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Characterization of Photosystem I in the Red Alga Porphyridium purpureum
KOSMAS, Panagiotis
Photosynthesis, a fundamental biochemical process on planet Earth, has passed through several steps during the course of evolution since beginning of time. Red algae represent a primitive stage of evolution of photosynthesis in eukaryotes, hence they are important for our understanding of functioning and evolution of photosynthetic energy conversion in general. In the current work, isolation and characterization of the supramolecular complex consisting of photosystem I with three antenna complexes from the red alga Porphyridium purpureum is presented. The purification consisted of sucrose gradient centrifugation followed by anion exchange and size-exclusion chromatography. The composition of the PSI-antenna supercomplex was confirmed using electrophoresis and reverse-phase chromatography. Function of the isolated supercomplex was further investigated using time-resolved absorption and fluorescence spectroscopy.
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The use of HPLC technique in the analysis of phenolic compounds in plant material of fruit trees
Adamcová, Anežka ; Šatínský, Dalibor (advisor) ; Sklenářová, Hana (referee) ; Vávra, Radek (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Mgr. Anežka Adamcová Supervisor: Prof. RNDr. Dalibor Šatínský, Ph.D. Title of the Dissertation Thesis: The use of HPLC technique in the analysis of phenolic compounds in plant material of fruit trees Although fruits are well studied and known for their high content of antioxidants and positive health benefits, the main aim of the presented dissertation thesis consists in the characterization of biological active compounds in individual parts of fruit tree material (leaves, bark, buds, flowers, and wood chips). The theoretical part is focused on fruit trees and the characterization of phenolic compounds. Their properties and the effects based on current scientific studies are shown. A large part of thesis is dealing with the chromatographic methods for the separation of phenolic substances including the current trends. The extraction process is also included. The practical part deals with the validation and development of screening HPLC-DAD method for the analysis of monitored phenolic compounds (phloridzin, phloretin, chlorogenic acid, quercitrin, rutin, arbutin, 1,5-dicafeoylquinic acid, and 3,5-dicafeoylquinic acid) in individual parts of fruit trees. The main objective of this thesis is to...
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Determination of methanol derivatives in drying oils
Horká, Hana ; Hraníček, Jakub (advisor) ; Janoš, Pavel (referee)
The formation of metal carboxylates is a major problem for historical and contemporary paintings, and therefore understanding the mechanism of their formation is a crucial step for developing appropriate conservation strategies and preserving the world's cultural heritage. The determination of reactive components in drying oils is an important step towards understanding the reaction mechanism. This work is aimed at developing a simple and rapid methodology for HPLC-DAD determination of low molecular weight degradation products of traditional drying oils, namely methanol, formaldehyde and formic acid. In this work, QuEChERS conditions for the extraction of methanol, formaldehyde and formic acid from various oil samples (olive, walnut, poppy and linseed oils) were optimized followed by purification of the samples from the oil matrix using a commercial d-SPE sorbent (Z-Sep/C18) and two nanostructured sorbents, CeO2 and ZrO2.
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Determination of non-esterified fatty acids in plasma
Lacinová, Petra ; Pacáková, Věra (advisor) ; Staňková, Barbora (referee)
The determination of non-esterified fatty acids (NEFA) in plasma is possible by many routine biochemical methods. For more detailed metabolic studies it is required to analyse the fatty acid (FA) profile. The NEFA profile determination is usually performed by the capillary gas chromatography (GC), but the sample preparation is not uniform. NEFA may be isolated by the extraction process or by the preparative thin-layer chromatography (TLC). The aim of this study was to compare these two separation procedures. The samples of pooled plasma from volunteer donors were analyzed by capillary GC after previous separation procedure by the TLC and the liquid-liquid extraction. The results were compared by the t-test for both the absolute concentration of individual FA provided by the internal standard (margaric acid) method and the relative abundance (Rel%). The reproducibility of the results was significantly better for the liquid-liquid extraction method than the TLC. The relative standard deviations (RSD) of the FA groups (unsaturated, monosaturated and polysaturated) were from 4.3 to 11.3 % vs. 8.6 to 33.8 % for the relative abundance and from 16.5 to 25.5% vs. 15.4 to 47.4% for the absolute concentration. For each FA, which were represented by more than 0.1 Rel %, RSD ranged from 4.5 to 47.8% vs. 9.5 to...
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Optimization of microextraction technique for analysis of water by chromatographic methods
Otrubová, Lucie ; Čabala, Radomír (advisor) ; Sobotníková, Jana (referee)
New liquid phase microextraction (LPME) has been optimized. Plastic adapter with tapered tip was used as extraction device. Concentration of analytes was determined by GC-MS. Face centered design was used to optimize LPME and the results were evaluated by Minitab 16 programme. Volume of sample, volume of extraction solvent, stirring rate, extraction time, ionic strength and time after microextraction were optimized. Response of system was defined as the sum of the peak relative areas. Toluene, tetrachlorethylene, ethylbenzene, xylenes, mesitylene and naphthalene were the studied analytes. Methylhexadecanoate was used as an internal standard. Optimal system conditions were as follows: 20 mL sample volume, 300 μL extraction solvent volume, 20 minutes, stirring speed 700 rpm, no addition of salt, and time after microextraction 3.5 minutes. Determination coefficient was 0.9700 and the lack-of-fit was insignificant which indicated good agreement of the model with the experimental data. The preconcentration factor ranged from 26 (tetrachlorethylene) to 39 (xylenes). The microextraction yield was varied from 39.5% (tetrachlorethylene) to 59.1% (p-xylene and m-xylene). The new method was tested by analyzing real samples of tap water, river water and water from the sewage treatment plant. The samples were...
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