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Developement and optimalization of methods for extraction of selected compounds from Hypericum perforatum
Chytil, Dalibor ; Štursa, Václav (referee) ; Pořízka, Jaromír (advisor)
Bachelor's thesis deals with the optimization of processes for extraction of phenolic compounds from St. John´s Wort (Hypericum perforatum). The theoretical part of the work focuses on characterization of this herb, its botanical classification, traditional use, chemical composition and curative effects. Increased attention is further devoted to a theory of preparation of samples for extraction of phenolic compounds and suitable techniques for extraction and characterization of phenolic compounds from extracts of St. John´s Wort. The experimental part of the thesis deals with application of theoretical knowledge. When optimizing the emphasis was on influence of solvent (water acified to pH 4, pH 5, pH 6), influence of extraction technique (ultrasound extraction, subcritical water extraction, maceration) and influence of extraction period (10, 20, 30 min). Extracts were subjected to determination of selected parameters (total phenolic content, total flavonoid content, antioxidant activity) in leaves of St John´s Wort harvested in June 2016 using UV-VIS spectroscopy. The used extraction techniques are significantly different from each other.
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The effect of abiotic elicitors on secondary metabolites content in plant cultures in vitro - I.
Teplá, Klára ; Tůmová, Lenka (advisor) ; Siatka, Tomáš (referee)
Plants are a source of a wide range of secondary substances, which due to their effects find use in many areas of focus. By a method called elicitation, we can achieve their higher and thus more efficient production. This diploma thesis aimed to determine whether the abiotic elicitor 2-(4-chlorophenyl)-N-(5-chloropyridin-2-yl)acetamide can positively affect the production of the flavonoid hyperoside in callus and suspension culture of Hypericum perforatum L. The elicitor was added to the in vitro cultures in three concentrations: C1 = 3,571.10-3 mol/l; C2 = 3,571.10-4 mol/l and C3 = 3,571.10-5 mol/l. A sample was taken at regular intervals after 6, 24, 48, 72 and 168 hours of elicitor treatment. Control samples were taken after 24 and 168 hours. The content of hyperoside produced was subsequently determined using High Performance Liquid Chromatography. Simultaneously, the amount of hyperoside released into the nutrient media of both plant cultures was also monitored. Maximum hyperoside production was recorded in suspension culture after 6 (17,7 µg/g DW) and 48 hours (3,69 µg/g DW) of elicitor treatment with the lowest concentration of C3 (3,571.10-5 mol/l). The content of hyperoside in the first case was 1770 % higher compared to the control sample. There was a significant release of hyperoside...
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The effect of pyrazine derivatives on secondary metabolites content in plant cultures in vitro II.
Hanzlíková, Tereza ; Tůmová, Lenka (advisor) ; Karlíčková, Jana (referee)
The main purpose of the theses was to find if the abiotic elicitor 2,4,6-trimethyl-N-(pyrazine- 2-yl)benzenesulfonamide has any influence on the secondary metabolites production in callus and suspension cultures of Hypericum perforatum L. The cultivation was taking place in the Murashige a Skoog (MS) nutrient medium enriched by the growth regulator - alpha-naphthyl acetic acid at the concentration of 1 mg.L-1 . The elicitor was added to the cultures at the three levels of concentration: c1= 100.0 mg/100 ml; c2= 10.0 mg/100 ml; c3= 1.00 mg/100 ml. The individual samples were taken after 6, 24, 48, 72 and 168 hours of the elicitor application. The control samples without the elicitor were taken after 6, 48 and 168 hours. The collected samples were dried and subsequently transformed into methanol extracts in order to determine secondary metabolites content (rutin, hyperoside and quercetin) by HPLC method. Release of these metabolites into nutrient media was also subject of this observation. The elicitation has influenced production of the secondary metabolites, particularly in callus cultures, wherein several statistically significant values, characterizing increase in their production, were measured. The highest content of rutin (0.169 mg.g-1 DW) was determined in callus culture after 168 hours when...
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Hypericin production in tissue cultures of Hypericum perforatum
Sazama, Pavel ; Martin, Jan (advisor) ; Kašparová, Marie (referee)
Production of hypericine in explantate cultures of Hypericum perforatum Pavel Sazama Diploma thesis Charles university in Prague, Faculty of Pharmacy in Hradec Králové, Pharmacy Key words: St. John's Wort, precursors, hypericine, flavonoids, acetate, cinnamic acid, cinnamate, tyrosine, shikimic acid The goal of this work was to affect the production of hypericin (naphto-dianthrone), hyperoside and quercitrin (flavonoids) in the suspensional Hypericum perforatum explantate cell cultures. The method of precursor feeding was used. The precursors of naphtodianthrones and flavonoids were added into the medium in final concentrations 10 mg.l-1, 50 mg.l-1 and 100 mg.l-1. Samples were taken after 72 and 168 hours after adding the precursor. Potassium acetate, cinnamic acid, sodium cinnamate, tyrosine and shikimic acid were used as precursors. Cultures were cultivated on the Murashige and Skoog medium with the addition of the growth stimulator α-NAA. Concentration of hypericin, hyperosid and quercitrin was measured by HPLC analysis. The highest influence on the production of hypericin in cultures was detected after adding of tyrosine. The concetration of hypericin in cells raised in this experiment from 0,003% (control culture) to 0,03% (culture with tyrosine concen-tration of 100 mg.l-1). This...
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Developement and optimalization of methods for extraction of selected compounds from Hypericum perforatum
Chytil, Dalibor ; Štursa, Václav (referee) ; Pořízka, Jaromír (advisor)
Bachelor's thesis deals with the optimization of processes for extraction of phenolic compounds from St. John´s Wort (Hypericum perforatum). The theoretical part of the work focuses on characterization of this herb, its botanical classification, traditional use, chemical composition and curative effects. Increased attention is further devoted to a theory of preparation of samples for extraction of phenolic compounds and suitable techniques for extraction and characterization of phenolic compounds from extracts of St. John´s Wort. The experimental part of the thesis deals with application of theoretical knowledge. When optimizing the emphasis was on influence of solvent (water acified to pH 4, pH 5, pH 6), influence of extraction technique (ultrasound extraction, subcritical water extraction, maceration) and influence of extraction period (10, 20, 30 min). Extracts were subjected to determination of selected parameters (total phenolic content, total flavonoid content, antioxidant activity) in leaves of St John´s Wort harvested in June 2016 using UV-VIS spectroscopy. The used extraction techniques are significantly different from each other.
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