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Micro-electromembrane extraction across multiple aqueous and organic phases for selective pretreatment of raw biological samples
Kubáň, Pavel ; Boček, Petr ; Seip, K. F. ; Gjelstad, A. ; Pedersen-Bjergaard, S.
Micro-electromembrane extraction (μ-EME) across multiple aqueous and organic\nsolutions was developed for selective extractions of various analytes from untreated\nbiological samples. The novel approach requires μL volumes of samples and organic\nsolvents, which form consecutive immiscible plugs in a transparent polymeric capillary\nand the extraction process is accelerated by application of d.c. voltage. Number of the\nplugs, their composition, volume and sequence in the extraction capillary can be chosen\nspecifically for each particular application and high variability and selectivity of the\nextraction process can be achieved. In this contribution, we demonstrate suitability of\na five-phase μ-EME system for simultaneous extractions of cations and anions from\nurine and for selective extractions of basic analytes based on their acid-base strength\nfrom plasma.
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Capillary electrophoresis with contactless conductivity detection for rapid determination of meldonium in urine
Šlampová, Andrea ; Kubáň, Pavel
A simple, rapid and cheap capillary electrophoresis-capacitively coupled contactless\nconductivity detection (CE-C4D) method for determination of meldonium (MEL) in\nurine samples was developed. Sample pretreatment was minimized to dilution of urine\nsamples with DI water and MEL was determined by CE-C4D in background electrolyte\nconsisting of 2 M acetic acid (pH 2.3) in ng mL−1 to mg mL−1 concentration range.\nSample-to-sample analysis time was less than 4 min, limit of detection and\nquantification was 0.015 and 0.05 μg mL−1, respectively, and the method showed\nexcellent linearity (r2 ≥ 0.9998), recovery (97.6–99.9%) and intraday as well as interday\nrepeatability (RSD ≤ 4.7%).
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A novel nanospray liquid junction interface for versatile CE-MS
Křenková, Jana ; Haselberg, R. ; Somsen, G. W. ; Foret, František
In this work we present a new polyimide-based liquid junction nanospray interface, combining the advantages of sheath flow and sheathless designs. The liquid junction interface was prepared as a hybrid microfabricated plastic liquid junction sprayer allowing to perform analysis in standard (30-100 micro m ID) separation capillaries. The interface incorporates a self-aligning port for coupling of the separation capillary as well as ports for automated flushing of both the liquid junction and the separation capillary. The interface permits performing CE both with and without electroosmotic flow. The use of large bore capillaries allows injection and preconcentration of larger sample volumes, favoring detection sensitivity. Different compositions of the spray liquid and BGE can be used when needed.
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Determination of content substances for selected species and varieties of roses.
Slavíková, Eva ; Sus, Josef (advisor) ; Súkeníková, Tereza (referee)
This bachelor's work is focused on comparing the most significant content substance, vitamin C, in fruits of chosen rose kinds species and varieties. Herbal products are often seen by people as products of "second choice", when failure of allopathic treatment occurs, which is focused especially on suppressing symptoms of illness, but it ignores health of the human body as whole. That's why are herbal products often alternative for therapy of chronic diseases. The base of this work is extensive literary research and results of laboratory measurings. Literary part of this work includes description of botanical classification of rose plants. There is stated basic characteristic of individual rose species and their morphological features. In following chapters is composed overview of effective substances in rose fruits and their effect on human health. In practical part of this work are included results of laboratory tests for determining ascorbic acid content for chosen samples of rose species and it also includes results of weighing and measuring of fruit samples of chosen rose species. There are also stated chosen rose species and shortly their origin and description, in the beginning of practical part. For practical part were chosen samples of species. Results of practical part are compiled into statistic tables and graphs and it confirms high content of ascorbic acid in rose fruits. This content is in range of cca 300 - 900 mg.100 g-1. Due to growing interest in alternative treatment options and "clean" natural product, it can be assumed, that many plants of Rosaceae family will keep its place in treatment practice. Those plants will be used not only for treatment of acute and chronic diseases, but also for cosmetic products production, thanks to its large effects and process options.
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Development of electronisation of local administration of Zábřežsko micro region in relation with e-government
Filipová, Lucie ; Ulman, Miloš (advisor) ; Martin, Martin (referee)
Thesis "Development of an electronisation of local administration of Zábřežsko micro region in relation with eGovernment"examines the issues of the development of electronisation of autonomy in relation to legislation and eGovernment. The topic of the public administration has been increasingly discussed in recent years not only in the context of increasing demands for processing speed and information, but also to facilitate transparency and sub-processes in public administration. Theoretical part describes the role of eGovernment in the public administration, the process of electronisation of public administration and individual services of eGovernment (Czech POINT, data boxes, basic registers, communication infrastructure of public administration), which are being in place with municipalities. The practical part evaluates the situation of software and hardware equipment, legislatively bound on a contract software and service of eGovernment on a sample of villages Zábřežsko. On the basis of the results of the questionnaire researches are determinated recommendations for sample of 20 micro region Zábřežsko.
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Nutritional analysis and optimization of breeding of selected species of edible insects under conditions of the Czech Republic with regard to human health
Adámková, Anna ; Kouřimská, Lenka (advisor)
All over the world, the edible insects are considered a highly nutritious food with high protein and fat content. However, the nutritional value of insect is not constant. It can be affected by species, developmental stage, rearing technology or nutrition.
Therefore, this thesis was aimed at obtaining the selected nutritional value of edible insects. Analyses were focused on the determination of the crude protein content, fat content, fatty acid profile and sterols in selected species of edible insects, which can be commonly reared in the Czech Republic. At the same time we also analysed samples of insects reared on the island of Sumatra to evaluate the influence of the climate on the nutritional value. The main aim of the thesis was the determination of optimal breeding conditions, developmental stages and feed rations for obtaining good production of insects with nutritional properties suitable for human nutrition. The analyses showed a high nutritional value of selected insect species, but also confirmed the significant differences in the content of individual nutrients between different species depending on the climate conditions and developmental stage. Comparing the fat and crude protein content in edible insects and other conventional sources of meat it has been found, that the examined insect is similar to beef concerning the fat and crude protein content. The results obtained are the basis for determining the appropriate rearing conditions and developmental stages for obtaining insect with the desired nutritional properties for human nutrition.
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Freezing technology of bull sperm in relation to its survivability and fertilization ability
Doležalová, Martina ; Stádník, Luděk (advisor) ; Jiří, Jiří (referee)
The aim of optimalization the insemination doses production is to provide the highest fertilization ability of spermatozoa during the demanding proces of processing fresh semen and its subsequent cryopreservation. Temperature changes causes spermatozoa damage during the cooling and freezing. Spermatozoa is exposed to cold shock and many others limiting factors, which leads to cell death and therefore to decline of fertilization ability of thawed insemination doses. For increasing spermatozoa resistance, exactly the plasma membrane resistance against cold shock was fraction of egg yolk LDL cholesterol (low density lipoprotein) at various concentrations into the comercially produced diluents added. It is believed that LDL acts possitively to plasma membrane and helps to maintain the fertilization ability of spermatozoa after thawing. Following step in the proces of insemination doses production is slow cooling of diluted semen and equilibration, when the straws are store at cooling box for 30 minutes to 240 hours. This period is necessary to penetrate of certain diluent components into the spermatazoa also maintain the balance between their intracellular and extracellular concentration. Also important is subsequent freezing temperature gradient of insemination doses. The most suitable freezing method is based on computer controlled temperature decline in freezing chamber which allows the precise control of ice crystals formation that could tear and kill the cell.
During 2012 to 2016 was repeatedly collected semen from the group of breeding bulls (n = 27, Holstein and Czech Fleckvieh breed) at AI centre. Semen which fulfill the standard entrance conditions in first step was evenly into several parts divided. For dilution the three types of comercially diluents AndroMed, Bioxcell and Triladyl with and without LDL addition were used. Into the diluents AndroMed and Bioxcell the concentration of LDL 4 %, 6 % and 8% into the dilent Triladyl 6 %, 8 % and 10 % was added. Diluted semen was filled into the glass capillares with volume 0,1 ml and temperature +4 °C. Subsequently the sample was placed to cold bath (0°C) for 10 minutes. Then the volume of capillare with physiological solution (37 °C) was mixed and for next 120 minutes was incubate. The effect of cold shock to proportion of live spermatozoa was evaluated by using Eosin and Nigrosine staining technique during heat test of spermatozoa survivability after spermatozoa heating and after 120 minutes of incubation.
The more suitable semen diluents which provide the higher spermatozoa resistance against cold shock were AndroMed and Bioxcell. Together the possitive effect of LDL addition into the diluents to lower decrease of proportion of live spermatozoa during heat test was found (P<0.05). The most suitable LDL concentration which had a favorable influence at spermatozoa resistance against cold shock was 6 % in diluent Bioxcell. Values of the proportion of live sperm were higher at the beginning of the heat test (+1.31% to + 3.2%) and after 120 minute incubation (+5.82% to +8.41%) compared to other diluents with and without addition of LDL.
In the next step the process of equilibration was optimized, is an important part of insemination doses production. The effect of the length of equilibration for subsequent fertilization ability of spermatozoa was evaluated using spermatozoa motility based of CASA and proportion of live spermatozoa after thawing and during heat survival test lasting 120 minutes (37 ° C). Suitable semen was diluted by comercially used diluent AndroMed based on soya lecithin, filled into the straws (0.25 ml), cooled and equilibrated in cooling box for 30, 120 and 240 minutes and freezed in programmable freezing box applying four types of freezing curves differing in temperature rate decline. There was used standard and by producer recommended 3. phase freezing curve, then 2. phase freezing curve, and 3. phase freezing curve with slower as well as rapid decline of temperature rate in freezing chamber, compared with standard freezing curve. The highest spermatozoa motility was found using 240 minutes of equilibration by +2.72% and +4.58% compared to other lengths of equilibration (P <0.05 to 0.01). The highest proportion of live spermatozoa was found using 120 minutes of equilibration (+6.87 % and +8.68 %). The highest average spermatozoa motility during heat test after thawing was achieved by using 2. phase freezing curve (from +2.97% to +10.37%, P <0.05), also in the proportion of live spermatozoa (from + 4.37% to +8.82%, P <0.01). When evaluating interaction between the length of equilibration and freezing curve (standard 3. phase and 2 . phase freezing curve), the highest average spermatozoa motility and proportion of live spermatozoa using 240 minutes of equilibration by both freezing curves was reached, there was no statistically significant differences. As well as, in all evaluated parts of this study the individual differences between ejaculate of bulls and within semen from one bull (P <0.05) as secondary effect were found.
To maintain good fertilization ability of semen during cryopreservation is necessary to increase the spermatozoa resistance against cold shock using addition of correct concentration of LDL into the commercially used diluents AndroMed and Bioxcell. Subsequently the fertilization ability of insemination dose is influenced by cooling, the length of equilibration and freezing. The length of equilibration 120 minutes and more as well as gentle way of freezing according to freezing curve, which ensures a gradual decrease of temperature in freezing chamber provided the higher average spermatozoa motility and proportion of live spermatozoa.
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