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Extraction of Tall Soap by Use of Selected Organic Solvents
Rousková, Milena ; Heyberger, Aleš
The aim of this study was to compare yields of phytosterols by use of liquid-liquid-extraction when several extraction solvents, recommended in literature sources, were used. Hexane, limonene, isobutanol, methyl isobutyl ketone and methyl tert-butyl ether were tested. These solvents differ in polarity. Largest yields of total phytosterols (67%) and especially of the beta sitosterol (89%) were reached when isobutanol was used. Disadvantages of most of the tested solvents were formation of the undesirable solid interphase, which could not be removed, their partial solubility in water phase and/or their high boiling point.
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Novel protease in the reduced mitochondria of Giardia intestinalis.
Vondráčková, Pavlína ; Doležal, Pavel (advisor) ; Hrdý, Ivan (referee)
The proteom of Giardia mitosomes is represented by twenty proteins, which are imported from the cytoplasm, as mitochondrial genome is missing. The N-terminal sequences, which target proteins to mitosomes are removed upon translocation and the remaining protein is folded into its native comformation. Out of three types of mitochondrial processing peptidases only functional half of dominant matrix peptidase is present in the mitosomes of G. intestinalis - feature unique among all eukaryotes. Based on the proteomic analyses, we identified novel putative mitochondrial peptidase (peptidase X) in G. intestinalis mitosomes. Overexpressing the protein with hemagglutinin tag we confirmed its mitosomal localization. Using specific polyclonal antibody combined with carbonate extraction and protease protection assay we have specified the distribution of the protein in the mitosomal subcompartments. Based on these results, peptidase X is anchored in the inner mitosomal mambrane via its C-terminus facing the mitosomal matrix. Peptidase X has mitochondrial targeting sekvence on the N-terminus, which is also capable of targeting protein into yeast mitochondria. By using bioinformatic tools, we are trying to detect structural similarities of peptidase X to other proteins in order to identify its substrate specifity and thus...
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Solid-phase extraction and its miniaturization by method Lab-On-Valve for determination of pharmaceutical substances
Jelínková, Iva ; Chocholouš, Petr (advisor) ; Vlčková, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical chemistry Candidate: Iva Jelínková Supervisor: PharmDr. Petr Chocholouš, Ph.D. Title of diploma thesis: Solid-phase extraction and its miniaturization by method Lab-On- Valve for determination of pharmaceutical substances This thesis deals with the development of the solid-phase extraction (SPE) method with the use of microextraction by packed sorbent (MEPS) for the determination of vitamins A, D, E and the possibility to automate this method. The determination was based on absorption of vitamins on microcolumn MEPS, interfering components were removed by washing solution (dilute acetic acid of pH 3) and extraction was performed by eluent solution (100% ACN). Detection was made by UV spectrophotometer at wave lengths of absorbing maximum for vitamin A - 325 nm, for vitamin D - 265 nm, for vitamin E - 295 nm. The method was optimized. There was developed a program, where concentrations of individual vitamins, dosing volumes, flow speed and composition solvents were tested. After the optimization method there was performed the extraction and the subsequent determination of blood plasma fortified with vitamins A, D, E by the sequential injection chromatography method. Recovery of the method for vitamin A was...
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Dental hygiene of pre-school children in the kindergartens in the district Jindřichův Hradec
ŠTONCNEROVÁ, Helena
The theme of dental hygiene of preschool children is a very current issue nowadays as ever growing importance is being ascribed to it. Regular and thorough removal of microbial dental plaque using appropriate technique plays an irreplaceable role in the prevention of dental caries, along with regular appointments with a dentist. The baccalaureate paper briefly maps the history and status quo of dental medicine, focusing primarily on the state of oral health of preschool children and prevention of dental caries. The theoretical chapter contains a description of teeth, the difference between temporary and permanent dentition, correct tooth cleaning technique, the frequency of tooth cleaning, and dental hygiene aids. The objective of the baccalaureate paper was to find out how and how often the nursery schools in the former district of Jindřichův Hradec attend to the dental hygiene of their pupils. Whether they are acquainted with the correct technique of tooth cleaning, the right grip of the tooth brush, and whether the subject of dental hygiene forms part of specialized lectures in these nursery schools were themes of my interest, too. Another part of my paper contains a draft of an educational program about dental hygiene, based on the results of a questionnaire that was part of my survey, with the aim of devising methodological instructions for preschool teachers as a way of promoting the adoption of proper habits in dental hygiene amongst preschool children.
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In vitro phytoextraction of non-steroidal antirheumatics
Houdková, Barbora ; Břichnáčová Habartová, Věra (referee) ; Smrček, Stanislav (advisor)
The most important question in the environmental research is presently the occurrence of the residues of pharmaceutical substances in ecosystem. These residues of drugs and their metabolites are entering our ecosystem by sewage water and lot of them are considered to be endocrine disruptors. The rhizofiltration is a possible method for the removal of this contamination. This work has researched the posibility of the phytoextraction of massively used non-steroidal anti-inflammatory drugs (NSAID) like diclofenac, naproxen and ibuprofen by Helianthus annus, Zea mays and Pisum sativum plants.The course of the phytoextraction of these substance in in vitro sterile model was compared to the extraction of their mixtures and we proved the reciprocal influence of these substance to the root systém. The presence of diclofenac or ibuprofen in Helianthus annuus plants increased the extraction of naproxen from the cultivation medium, whereas ibuprofen and diclofenac mutually decreased the phytoextraction efficiency. The experiments with Zea mays showed inhibitive trends whereas the experiments with Pisum sativum showed no influence in extraction of the mixture in comparison to the trials with only one substance. The mutual influence of the extraction efficiency of the solution with more substance is probably...
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Study of Pharmaceuticals Degradationby Advanced Oxidation Processes
Bílková, Zuzana ; Čáslavský, Josef (referee) ; Beklová, Miroslava (referee) ; Zachariášová,, Milena (referee) ; Vávrová, Milada (advisor)
At present, the issue of occurrence of female sex hormones, estrogens and progestogens, in aquatic ecosystems is often discussed by experts and the general public. These substances of steroid structure can be difficult to remove completely by conventional wastewater and drinking water treatment technologies. In given context advanced oxidation processes based on in situ generation of highly reactive hydroxyl radicals can be a suitable technique. This thesis deals with the study of kinetics and degradation products of photocatalytic decomposition of seven female sex hormones (estrone, -estradiol, estriol, ethinylestradiol, diethylstilbestrol, progesterone and norethindrone). Experiments were conducted in a laboratory glass reactor, which was equipped with an energy efficient UV-A LED light source (365 nm emission wavelength) and an immobilised photocatalyst in a form of TiO2 five-layer film deposited on glass. Model samples of water with the initial hormone concentration of 1 mg·L-1 were used and the degradation process was monitored by an HPLC-MS method. In the given system all compounds of interest except estriol had very significant tendency to be adsorb. In the case of estriol the formal rate constant of photocatalytic decomposition was determined based on the Langmuir-Hinshelwood model for two different initial concentrations, 0.5527 hour-1 (1 mg·L-1) and 0.1929 hour-1 (5 mg·L-1), and by comparison of these values it was found that the higher degraded compound concentration, the slower decomposition (fivefold increase of the initial concentration resulted in the constant decrease to almost one-third). Moreover nine degradation products of estriol photocatalytic decomposition were recorded and their structure was designed based on mass spectra. In the second thematic part of the thesis attention was paid to development of a SPE-HPLC-MS method for simultaneous determination of female sex hormones in water ecosystems, with emphasis on an extraction part optimization. The final samples treatment process included besides extraction with Supel™ Select HLB 200 mg SPE cartridges also mechanical impurities removal, hormones extraction from solids trapped on filtration material, sample acidification and extract purification with Supelclean™ ENVI-Florisil® cartridges. Optimised method was used for determination of female sex hormones in two Brno rivers, Svitava and Svratka. In the most cases the concentration was below the detection or quantification limit.
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Application of gas chromatography for determination of drug residuals
Sýkora, Richard ; Lisá, Hana (referee) ; Vávrová, Milada (advisor)
Analgesics are nowadays widely used group of drugs in human and also in veterinary medicine. Residuals of these drugs enter surface waters via discharges from the waste water treatment plants as the result of insufficient efficiency of their removal during cleaning process. In this bachelor thesis the possibilities of the analgesics residuals determination in waters using derivatization followed by gas chromatography with mass spectrometric detection were experimentally evaluated. Individual derivatization procedures were compared too. Several derivatization agents were tested and experimental parameters for selected analgesics were optimized (reaction time and temperature, amount of derivatizing agent). Further, several procedures of isolation of target compounds from water based on Solid Phase Extraction were verified. In all cases, gas chromatography with TOF-based mass spectrometric detection acts as final analytical method.
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Periodontal Disease of Dogs
Škutová, Šárka ; Barták, Miroslav (advisor) ; Olekšáková, Tereza (referee)
Periodontal disease (PD) affects periodontium and thanks to its prevalence it is considered to be one of the most occurred diseases in small animals and even in human. The uppermost etiological agens presents bacterial plaque. Bacterial plaque strongly accumulate if it is not eliminated and it might be the cause of development of gingivitis or even periodontitis over time.
The anatomy itself is relatively large, since the oral cavity is comprised of many kinds of tissues and a detailed description thereof would cover 70 pages at least. That is why the main aim of this work was to attempt to select the most basic information only.
Due to dividing of anatomical structures into bones, soft tissues, salivary glands and teeth, the individual components of supporting apparatus (periodontium) are not described in chapter called ,,periodontium,,. Here is explained its purpose and a description and function of periodontal ligament, which has not been included above. Particular attention was paid to the teeth. Determining the type and the number of individual teeth, the knowledge of their structure and the number of roots, all of that is important when it comes to a tooth extraction. The knowledge of tooth marking, dental formulas, identification systems and odontogenesis itself is equally important.
As for each disease, the etiology is the essential part for understanding the formation of disease. Bacterial plaque, which adheres to enamel of teeth is regarded (as already mentioned) as initial etiological agens. It can mineralize in tartar in varying degrees depending on every individual subject (animal). There are involved other risk factors at onset of the disease such as age, breed, number and line-up of the teeth, their pathology, diet, but also some diseases.
PD can be classified in two basic forms (gingivitis, periodontitis). Gum inflammation, or gingivitis, is often denoted as initial and totally reversible stage of disease. It is possible that there may not be present any symptoms or just slight redness at the beginning. Later stage leads to swelling, pain and significant bleeding, whereas it can manifest in periodontitis if therapy is absent.
Periodontitis affects the entire periodontium and it is often called as an irreversible stage. Although thanks to guided tissue and bone regeneration it is possible to achieve at least slight improvement these days. In most patients disease passes into chronic form, causes them not only pain during eating, but may also result in various systemic diseases. A special form which affects younger individuals is called ,,juvenile periodontitis,,. Diagnosis is primarily based on the medical history and examination of the oral cavity. Tartar index, gingivitis index, grades of tooth mobility and other parameters, which helps to determine the extension of disease and stage can be obtained using the intraoral radiographic, visual examination and measurement by periodontal probe. All collected information can be used to devise recommended therapy.
The sole therapy depends on the state of the patient and the requirements of the owner. It consists of several steps. The essence is especially removal of any deposits of bacterial plaque and tartar, correction of teeth pathologies (smoothing rough surfaces) and tissues pathologies to eventual tooth extraction. As part of the therapy may also be chosen application of antibiotics and other preparations supporting wound healing. Many specialists have been developing new treatments over the past 10 years, but most of them are still not fully finished. These include the development of vaccines, photodynamic therapy, or guided tissue and bone regeneration, which were already mentioned.
An integral part of therapy is home dental care. Without consistent home care there is a high probability of relapse.
An important weapon against the rise of PD are various preventive measures. The most effective method of prevention is probably regular tooth brushing using a brush and a special paste made for animals. However, many owners prefer to avoid this method. They rather elect to feed dry diets, dental treats and other products which help to reduce the amount of plaque and tartar. Another option is a professional teeth cleaning performed by a veterinarian.
A survey via questionnaire consisted of 16 questions, which were aimed at awareness of the disease, age, sex, breed, size, diet, preparations which help to control amount of plaque and tartar, systemic diseases and especially the presence of PD, age at first hit by disease, the circumstances of diagnosis, stage, treatment and relapse.
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Study of Transfer of Musk Compounds in Abiotic and Biotic Components of Aquatic Ecosystems
Zouhar, Libor ; Hajšlová, Jana (referee) ; Sokol, Jozef (referee) ; Večeřa, Zbyněk (referee) ; Vávrová, Milada (advisor)
This work was focused on the determination of musk compounds in various components of the environment. Synthetic musk compounds are organic substances commonly used as fragrant constituents of detergents, perfumes, cosmetics and other personal care products (PCPs). These compounds contaminate the aquatic environment through municipal wastewater from sewage treatment plants. The result is accumulation of these analytes in the environment and their occurrence in food chain. Synthetic fragrances are commonly detected also in human body; these substances generally belong among relatively new persistent organic pollutants (POPs) because of their persistence, bioaccumulation, and potential toxicity. The aim of this work was to optimize the methods for the determination of selected synthetic fragrances in wastewater (municipal, industrial) and in fish muscle. These analytes were determined in wastewater [the wastewater treatment plant (WWTP) Brno – Modřice, cosmetic manufacturing plant] and in fish Squalius cephalus that had been caught in the river Svratka. The selected analytes (14 substances) originate from the group of nitromusk, polycyclic musk and linear musk compounds. Solid phase microextraction (SPME) [for wastewater samples] and pressurized solvent extraction (PSE) [for fish samples] were used for isolation of analytes from real samples. The identification and quantification of analytes were carried out by high resolution gas chromatography coupled with mass spectrometry detection (GC-MS). The results show that galaxolide (g.L-1) and tonalide (tenths of g.L-1) were polycyclic musk compounds which were found at the highest concentration level in influent and also in effluent samples from the WWTP Brno – Modřice. The most extended nitromusk compound was musk ketone (hundredths of g.L-1). Linear musk compounds were detected in all influent samples (g.L-1). The overall removal efficiency of synthetic musk compounds from wastewater in the WWTP varies within the following ranges: nitromusk compounds 34.4 % – 85.4 %, polycyclic musk compounds 36.7 % – 53.3 % and linear musk compounds 90.5 % – 99.9 %. On the basis of the results it can be stated that wastewater (effluent from the WWTP) is the primary source of musk compounds, especially of the polycyclic ones in the environment. It has been also shown that ultrafiltration used in a cosmetic plant is a highly efficient technology for removing nitromusk and polycyclic musk compounds from industry wastewater. The results from evaluation of the fish contamination by musk compounds show that galaxolide was the substance which reached the highest concentration levels (tens of g.kg-1 l.w.). The results corroborate ubiquitous occurrence of musk compounds in the environment and reduction of using of nitromusk compounds due to their potential toxic properties. It has been shown that the insufficiency of the purifying procedures used in the WWTP Brno – Modřice has a significant influence on the concentration level of polycyclic musk compounds (tonalide, galaxolide) in fish caught from the river Svratka.
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Use of Capillary Zone Electrophoresis for Determination of Selected Analgetics in Water
Čapka, Lukáš ; Opatřilová, Radka (referee) ; Márová, Ivana (referee) ; Vávrová, Milada (advisor)
From viewpoint of environmental analysis in the whole world became popular in the latest years the question of drugs’ breakthrough to the component of environment. These contami-nants belong to the biological active compounds, with different physical-chemical and biolog-ical properties and evince great tendency to bioaccumulation. They penetrate to the environ-ment because of their increasing of usage and wrong techniques of liquidation. The most often use drugs are preparations against pain – analgetics, and from this category there are non-steroidal anti-inflammatory drugs. The frequent usage of this compounds relate to their easy accessibility. From this large group of compounds was chosen for monitoring: diclofenac, ibuprofen, ketoprofen, salicylic acid, naproxen and acetaminophen; because they include in favorite preparations. The monitoring matrix was the wastewater from two waste water treat-ment plants (WWTP). The sampling was performed in inflow and outflow because the com-paring of concentration of selected contaminants and discovering of efficiency of removing the polutants reliance on treatment technology. For extraction of selected contaminants was used solid phase extraction (SPE) and for determination was used capillary zone electrophore-sis (CZE) with diode array detection (DAD). There was identified and quantified all of se-lected analgetics in inflow and so in outflow of WWTP. That means, this polutants infuse into surface water and then into other components of environment.
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