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Analysis and phytoextraction of fluoxetine in water ecosystem
Rathouská, Lubomíra ; Jelínek, Ivan (advisor) ; Soudek, Petr (referee)
At present time, the main interest of eco-chemical studies focuses on environmental fate of residues of typical representatives of various pharmacological groups. The goal of these studies is to develop analytical methods for their precise determination and methods for elimination or lowering of the residuals concentrations in ecosystem. In this work we studied the possibilities of pre-concentration of Fluoxetine (antidepressant) from various analytical matrices by two common SPE resins (reversed phase (RP, Varian) and DVB-N-vinylpyrrolidone copolymer (Oasis HLB, Waters)). Possible phytoextraction of Fluoxetin by sunflower (Helianthus annuus L.) and maize (Zea mays L.) was also studied in model systems.The Oasis Hlb columns with subsequent elution with acetone were found as practically the most suitable system for Fluoxetin pre-concentration. This method allows threefold use of the colmuns. The phytoextraction showed positive results. The yield of Fluoxetin phytoextraction by sunflower was in order of 30-40 % in five- days cultivation and 20 % by maize under the same conditions.These results show that Fluoxetine can be extracted from aqueous solutions, even with slightly lower yield compared to other pharmaceuticals.
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Detection of microorganisms in the soil using CARD-FISH (Catalyzed Reporter Deposition - Fluorescence in situ hybridization)
Krištůfek, Václav ; Kasalický, Vojtěch ; Chroňáková, Alica ; Elhottová, Dana ; Němec, Jan
CARD-FISH protocol (Catalyzed Reporter Deposition - Fluorescence in situ Hybridization) for detection of soil microbial community composition was successfully established in the Institute of Soil Biology. CARD-FISH is the relatively new molecular biology tool permitting to quantify distinct groups of microorganisms by hybridization of specific oligonucleotide probes. Compared to classic FISH-protocol the probe-attached enzyme amplifies the fluorescent signal and enables to detect nearly all microorganisms in soil or in organic compounds. As the first, we used this method to determine the bacterial community composition of a dated layer of guano-heap from Domica Cave (Slovak Carst National Park, Slovakia). We quantified the percentuel composition of Archaea, Eubacteria (.i.Planktomycetales, Verrucomicrobiales./i. included) and major bacterial groups (Actinobacteria, proteobacteria, .i.Cytophaga-Flavobacter./i.-Bacteroidetes, Sphingobacteria).
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NMR Analysis of the Substances Isolated from a Corydalis Cava Plant (Fumariaceae) and from an Eschscholtzia Californica Plant (Papaveraceae)
Dohnalová, Hana ; Kuneš, Jiří (advisor) ; Špulák, Marcel (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of inorganic and organic chemistry Candidate: : Mgr. Hana Dohnalová Consultant: Doc. PharmDr. Jiří Kuneš, CSc. Title of Thesis: NMR analysis of the substances isolated from a Corydalis Cava plant (Fumariaceae) and from an Eschscholtzia Californica plant (Papaveraceae) This Rigorous work is dedicated to the determination of the structure of unknown substances. The isolation of alkaloids from a Corydalis Cava plant (Fumariaceae) and from an Eschscholtzia Californica plant (Papaveraceae) is carried out successfully at the Departement of Pharmaceutical Botanic and Ecology of the Pharmaceutical Faculty in Hradec Králové. A large number of alkaloids of several structural types was isolated there. Some of them were passed to the Laboratory of the structure and interaction of the biological aktive molecules along with the request for the structure analysis. The main object of this Rigorous work was the identification of three of unknown subjects by the help of NMR analysis. The structures of unknown substances were identified on the basis of the NMR experiments (1 H - NMR, 13 C - NMR, DEPT, ghmbc, ghsqc, ghmqfcops a dpfgnoe). One structure was confirmed based on the comparison with the data provided in the subject publications....
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Interakce xenobiotik s MDR pumpami u S. cerevisiae
Hendrych, T. ; Zlámalová, Marta ; Gášková, Dana ; Pichová, Alena ; Sigler, Karel
The interactions of a number of xenobiotics with two main S. cerevisiae MDR pumps, Pdr5p and Snq2p were studied. The results showed that some compounds do not interact with MDR pumps whereas others affect one or another of the pumps, causing a loss of their ability to transport an indicator substrate out of the cells. This apparent inactivation of the pumps can be due to competition for transport with the indicator substrate or to actual (noncompetitive) inhibition of the pumps
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Enzymes of Candida tropicalis yeast biodegrading phenol
Koubková, Zuzana ; Stiborová, Marie (advisor) ; Turek, Michal (referee)
Effluents of industrial wastewaters from oil refineries, paper mills, dyes, ceramic factories, resins, textiles and plastic contain high concentrations of aromatic compounds, which are toxic to organisms. Degradation of these compounds to tolerant limits before releasing them into the environment is an urgent requirement. Candida tropicalis yeast is an important representative of eucaryotic microorganisms that are able to utilize phenol. During the first phase of phenol biodegradation, cytoplasmatic NADPH-dependent phenol hydroxylase of C. tropicatis oxidizes phenol to catechol. Catechol is in the second phase of biodegradative process oxidized to cis,cis-muconic acid by the reaction catalyzed with catechol-1,2-dioxygenase. In this diploma thesis we investigated the effect of the heavy metal ions on NADPH-dependent phenol hydroxylase and catechol-1,2-dioxygenase of C. tropicalis. Phenol hydroxylase was inhibited by Cu2+ and Pb2+ ions. Catechol dioxygenase was inhibited by all substances containing heavy metal ions (Fe2+ , Mn2+ , Cd2+ , Cu2+ and Pb2+ ), which were tested in this work. The most effective inhibition was produced by Pb2+ followed by Mn2+ , Cd2+ Fe2+ and Cu2+ ions. The higher sensitivity of catechol-1,2-dioxygenase to heavy metal ions might follow from the presence of histidine residue...
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Use of NMR in structural analysis of natural products
Dohnalová, Hana ; Špulák, Marcel (referee) ; Kuneš, Jiří (advisor)
This Diploma work is dedicated to the determination of the structure of unknown substances. A large number of alkaloids was isolated from an Eschscholtzia Californica plant (Papaveraceae) at the Department of Pharmaceutical Botanic and Ecology of the Pharmaceutical Faculty in Hradec Králové. Some of them were passed to the Laboratory of the structure and interaction of the biological aktive molecules along with the request for the structure analysis.The main object of this Diploma work was the identification of three of unknown subjects by the help of NMR analysis. The structures of unknown substances were identified on the basis of the NMR experiments (1 H-NMR, 13 C-NMR, DEPT, ghmqc, ghsqc, ghmqfcops a dpfgnoe). These structures were confirmed based on the comparison with the data provided in the subject publications. Now identified substances belong to isochinoline alkaloids. Namely, it is scoulerine and retikuline. The third substance was not specified in the subject publication.
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Diffusion parameters of brain tissue during ischemia and during status epilepticus
Šlais, Karel ; Syková, Eva (advisor) ; Otáhal, Jakub (referee) ; Rokyta, Richard (referee) ; Dezortová, Monika (referee)
Difusion of neuroactive substances in the extracellular space (ECS) is constrained by two factors: extracellular space volume and geometry. We have shown changes of diffusion parameters and extracellular concentrations of energetic metabolites and glutamate in two pathological states accompanied by significant ECS volume changes - in combined hypoxia/ischemia and in status epilepticus. In the model of hypoxia/ischemia, we have shown time courses of diffusion parameters, concentrations of glucose, lactate, lactate/pyruvate ratio and glutamate during hypoxic/ischemic insult and after reperfusion. The time course of glutamate extracellular concencetration in transient hypoxia/ischemia correlated well with time course of changes in diffusion parameters. The decrease in the ECS volume fraction can therefore contribute to an increased accumulation of toxic metabolites, which may aggravate functional deficits and lead to damage of the central nervous system. In the model of pilocarpine-induced status epilepticus our results show changes in the extracellular space diffusion parameters, K+, energy-related metabolites and glutamate during the initiation and first hours of the propagation. Our results also show that the first minutes after a pilocarpine injection are followed by an increase in extracellular K+ and a...
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Phytochemical study of Evolvulus alsinoides IV
Svačinová, Jana ; Karlíčková, Jana (advisor) ; Opletal, Lubomír (referee)
SOUHRN Provedla jsem výzkum petroletherového extraktu Evolvulus alsinoides L. Sloupcovou chromatografií jsem získala celkem 113 frakcí, z nichž pro další výzkum jsem zvolila frakce 3-4, 5-7, 15-16, 17, 36-40, které se jevily jako perspektivní. Sumární petroletherový (Pe) extrakt byl podroben testování na akutní toxicitu. Test akutní toxicity nebylo možno s extraktem adekvátně provést, jelikož podmínkou jeho provedení je rozpustnost vzorku ve vodě. U emulze Pe extraktu s vodou byl výsledek negativní. Pe extrakt vykazoval slabou antioxidační aktivitu. Výsledky testování na antioxidační aktivitu mohly být zkresleny vzhledem k malé rozpustnosti extraktu v použitém rozpouštědle. Z frakce 3-4 (1,3244 g) bylo získáno 0,02 g čistých krystalů bílé barvy a voskovité konzistence s teplotou tání 58-82 řC, které byly dle provedené TLC pravděpodobně směsí tří látek. Srovnávací TLC ukázala, že se nejednalo o kyselinu palmitovou a stearovou. Pomocí GC/MS bylo zjištěno, že krystalický podíl frakce 3-4 představuje směs látek. Jedná se o kyselinu 8-methyldekanovou, kyselinu olejovou, alifatický uhlovodík a polynenasycené mastné kyseliny, které nebylo možno pro jejich malé množství blíže specifikovat. Stanovení antioxidační aktivity u krystalického podílu frakce 3-4 bylo negativní. Frakce 5-7 (2,7028 g) poskytla 0,0052 g...
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