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Abiotic elicitation of explant culture of Trifolium pratense L.
Fiedlerová, Gabriela ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Abiotic elicitation of explant culture of Trifolium pratense L. Summary The objective of this dissertation was to observe the effect of the abiotic elicitors of mercuric chloride and cadmium chloride on the production of flavonoids, and also of the effect of the mercuric chloride elicitor on the production of isoflavonoids in the Trifolium pratense L. explant culture (variety DO−8). The cultures were cultivated in the Gamborg nutrient media with the addition of 2,4−dichlorphenoxyacetic acid (2 mg.l-1 ) and 6−benzylaminopurine (2 mg.l-1 ), in the temperature of 25 řC and during a 16-hours light/8-hours dark period. The quantity of flavonoids was determined spectrophotometrically according to The Czech Pharmacopoeia 2005. The quantity of isoflavonoids was determined by the HPLC method. We can see from the results of this dissertation that the concentration 10 μmol of both of the used abiotic elicitors had the greatest positive effect on the production of flavonoids. The largest quantity of flavonoids (0,576 %) was detected during the elicitation of cadmium chloride of a 10-μmol concentration and after 48−hour application when the quantity of flavonoids increased by statistically significant 294 % in relation to the control sample. The quantity of isoflavonoids was determined by the HPLC method during the...
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Alkaloids from Narcissus pseudonarcissus L. cv. Dutch Master and their cholinesterase and prolyl oligopeptidase inhibitory activity.
Farkašovský, Marek ; Chlebek, Jakub (advisor) ; Siatka, Tomáš (referee)
Farkašovský Marek: Alkaloids from Narcissus pseudonarcissus L. cv. Dutch Master and their cholinesterase and prolyl oligopeptidase inhibition activity. Rigorous thesis 2017, Charles university, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology. The content of this work was isolation of minimum two compound from alkaloid fraction ND 3 - 5/2, obtained by column chromatography of Narcissus pseudonarcissus L. cv. Dutch Master alkaloid extract. Preparation of extract and its column chromatography was performed by Mgr. Daniela Hulcová as a part of her doctoral studies. Two different alkaloids were isolated with preparative TLC method. Their structure was determined with optical rotation, MS (EI, ESI), 1H NMR and 13C NMR analysis. Obtained data were compared with facts with those in literature. The first isolated compound was lycorine type alkaloid O-acetylpluvine. Its inhibition activity compared to positive standards (galanthamine, huperzine A, physostigmine, berberine and Z-pro-prolinal) was mild - IC50 AChE: 684 ± 54 µM, IC50 BuChE: 603 ± 49 µM, IC50 POP ˃1000 µM. From the point of its cholinesterase and POP inhibition, this results make O-acetylpluvine unusable compound in treatment of AD. The second isolated compound was a new alkaloid, which has not been...
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Biological activity of plant metabolites. XV. Influence of alkaloids from Chelidonium majus L.on acetylcholinesterase
Nagyová, Jana ; Opletal, Lubomír (advisor) ; Siatka, Tomáš (referee)
Nagyová, J.: Biological activity of plant metabolites XV. Influence of alkaloids from Chelidonium majus L. on acetylcholinesterase, Rigorous work, Charles University in Prague, Faculty of Pharmacy Hradec Králové, 67 pp. Key words: Alzheimer's disease, alkaloids, Chelidonium majus, acetylcholinesterase, biological aktivity. Within the screening of plants that contain alkaloids inhibiting the activity of the human erythrocytic acetylcholinesterase Greater celadine (Chelidonium majus L., Papaveraceae) was studied. 41.8 kg of the whole dry plant (the aerial part with the roots) served for the isolation of alkaloids. Mixtures of pseudocyanides of benzophenanthridine alkaloids, phenolic bases, and the alkaloids precipitated from either chloroform-soluble or chloroform-insoluble chlorides were obtained from the purified ethanolic extract. My task was to separate benzophenantridine alkaloids from pseudocyanides. Using both column and thin layer chromatography chelerythrine as a chloride was isolated. The identity of the isolated alkaloid was determined by means of an authentic standard and by comparing its physico-chemical characteristics with the published data. The isolated compound inhibited the human erythrocytic acetylcholinesterase with IC50 5.89.10-6 M. Comparing its biological activity with that of...
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Basic phytochemical investigation of some taxons division Eumycota with respect to study of antioxidant activity
Mrvíková, Hana ; Opletal, Lubomír (advisor) ; Siatka, Tomáš (referee)
SOUHRN Tato práce si kladla za cíl provést základní fytochemický výzkum vybraných 14 taxonů hub oddělení Eumycota. Pojednávané druhy hub patří celkem do sedmi čeledí: Boletaceae, Hydnaceae, Pleurotaceae, Polyporaceae, Russulaceae, Strophariaceae a Telephoraceae. Nejpočetněji je zde zastoupena čeleď Russulaceae kam náleží osm hub ze sledovaných 14 druhů. Příprava extraktů pro účely v této práci spočívala jen v navážení potřebného množství již hotového lyofilizátu a přidání vhodného rozpouštědla. Práce se zabývala dvěma experimentálními základy. Tenkovrstvou chromatografií se skupinovými reakcemi a stanovením antioxidační aktivity pomocí DPPH testu s použitím metody SIA. Literární rešerže byla zaměřena jak na informace o obsahových látkách sledovaných taxonů tak na údaje o jejich biologické aktivitě a případné toxicitě. Z hlediska antioxidační aktivity se zdá být zajímavá houba Albatrellus ovinus. Ve studii 3- hydroxyneogrifolin a 1-formyl-3-hydroxyneogrifolin, látky obsažené v její plodnici, vykazovaly silnější antioxidační aktivitu než α - tokoferol nebo BHA. 14 Při použití metody SIA pro stanovení antioxidační aktivity DPPH testem byly použity koncentrace sumárních extraktů 0,1 mg/ml, 0,5 mg/ml, 0,25 mg/ml a 0,1 mg/ml. Standardem byl Trolox měřený při koncentracích 0,5 mg/ml, 0,1 mg/ml, 0,05 mg/ml a 0,025...
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The Influence of Elicitation on the Production of Suspension Culture of Trifolium pratense L.
Keřková, Adéla ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Adéla Keřková The Influence of Elicitation on the Production of Suspension Culture of Trifolium pratense L. Summary The paper examined the effect of four concentrations of cadmium chloride on the production of flavonoids and isoflavonoids by the Trifolium pratense L. suspension culture (variety DO-9 and SPRINT). The cultures were cultivated in the Gamborg nutrient media with the addition of 2 mg.l-1 of 2,4-dichlorophenoxyacetic acid and 2 mg.l-1 of 6-benzylaminopurine, at the temperature of 25 o C, 16-hr light/ 8-hr dark period. The elicited and the inspection samples underwent the photometric determination of flavonoids in accordance with the Czech Pharmacopoeia 2005 and the determination of flavonoids via the HPLC metod. The results show that the optimal effect of cadmium chloride on the production of flavonoids and isoflavonoids was manifestated after a 168-hour application of a concentration 0,1µmol (variety DO-9) and after 6-hour application (variety SPRINT).
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Alkaloids of the family Amaryllidaceae and their biological activity 1.
Klímová, Kamila ; Cahlíková, Lucie (advisor) ; Siatka, Tomáš (referee)
Mišáková K: Alkaloids of the Amaryllidaceae family and their biological activity 1. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology, Hradec Králové, 2014, 63 p. The aim of the diploma thesis was a preparation of alkaloidal extract from Nerine Bowdenii to izolation of 2 pure alkaloids by column and thin layer chromatography. Subsequently these two alkaloids were subjected to structural MS and NMR analysis and tested for biological activity against human cholinesterases (HuAChE and HuBuChE) and for antioxidant and cytotoxic activity. Isolated substances were identified as ambelline and undulatine alkaloids, which have been already described previously in this plant. In the test for determining cholinesterase inhibitory activity of ambelline and undulatine the following values were measured; for ambelline: IC50, HuAChE = 169 ± 7 μM a IC50, HuBuChE = 985 ± 25 μM, for undulatine: IC50, HuAChE = 23,52 ± 1,19 μM a IC50, HuBuChE > 1000 μM. Galanthamin (IC50, HuAChE = 1,71 ± 0,007 μM, IC50, HuBuChE = 42,30± 1,30 μM), huperzin A (IC50, HuAChE = 0,033 ± 0,001 μM, IC50, HuBuChE > 1000 μM) and eserin (IC50, HuAChE = 0,063 ± 0,001 μM, IC50, HuBuChE = 0,130 ± 0,004 μM) were used like a positive controls. In the test for...
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