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Effect of post-harvest processing on quality of Sambuci fructus
Brňáková, Lenka ; Spilková, Jiřina (advisor) ; Kašparová, Marie (referee)
Elderberry is very popular plant, that is due to its beneficial effects to human organism very reputable. The main content substances are flavonoids and anthocyanins. Fruits of this flower are available only seasonally (which is usual for most of flowers), so we have to search for alternative solution of getting them. One of these solutions is conservation. Nowadays there are lots of types of postharvest adjustment, so we have more opportunities to find an optimal concept for every one specific plant. The diploma thesis is focused on assessment of content of phenolic substances harvested in different parts of Hradec Králové. They were conservated at laboratory temperature, elevated (40řC, 60řC) and reduced (-18řC) temperature. Next part of thesis is focused on assessing of the content of anthocyanins in fruits, that had been conservated for 3 years in refrigerator. The theme was finding the best concept of postharvestal adjustment. Despite to the individual places of harvest, there were not mentionable differences found out in content of phenolic substances in fruits. Main differences in content of substances were caused by different temperature of conservating area. With the upper temperature the content of phenolic substances was lower. Storage in the fridge for 3 years did not have any influence...
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Study on the interaction of chemopreventive compounds and food born carcinogens with cytochrome P450 enzymes
Brabencová, Eliška ; Hodek, Petr (advisor) ; Koblihová, Jitka (referee)
The use of food supplements containing natural chemopreventive compounds increased in recent years. Some of the most popular chemopreventive compounds are flavonoids. Due to their natural origin, flavonoids are generally accepted as safe compounds. They exert antioxidant, anti-cancer and anti-inflammatory properties. However, flavonoids should be considered as foreign compounds (xenobiotics). Flavonoids interact with many enzymes, among the most important belong cytochromes P450 (CYPs), key enzymes of the first phase of biotransformation of xenobiotics (e.g. drugs, carcinogens). CYPs catalyze reactions leading mainly to detoxification of xenobiotics. However, some CYPs are involved in the activation of carcinogens, particularly CYP1A1 and CYP1A2 activate e.g. heterocyclic amines. Flavonoids might enhance the activation of carcinogens via induction of these CYPs or stimulation of their activities and hence, increase the risk of a cancer development. The thesis is focused on the influence of flavonoids and food carcinogens on the induction and activity of CYP1A1 and CYP1A2 in liver and small intestine of rats. For the purpose of this study, the small intestine was dissected into three parts: proximal (nearest to stomach), middle and distal. Western blotting was used for the evaluation of CYP...
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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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Biological effects of various hemp fractions
Vacková, Hana ; Skoumalová, Petra (referee) ; Němcová, Andrea (advisor)
Cannabis is the only plant which contains cannabinoids and thanks to these compounds it has enormous potential. This thesis deals with the analysis of technical hemp. Effects of cannabinoids and methods used for cannabis analysis are discussed in the theoretical part. The experimental part includes spectrophotometric characterization of cannabis, it´s antimicrobial effects and thin layer chromatography analysis. Three sorts of Cannabis sativa L. were analyzed, namely Finola, Fedora and Kompolti. Firstly, the content of polyphenols, flavonoids and antioxidant activity in prepared tinctures were determined. Moreover, antimicrobial test were performed using disk test and turbidity determination. Gram-positive and Gram-negative bacteria and yeast organism were tested. It was found that cannabis tinctures possess good antimicrobial effects. Some of them are comparable with synthetic antibiotics. Finally, thin layer chromatography enabled visualization of cannabinoids in prepared tinctures.
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Sea buckthorn (Hippophae rhamnoides L.) as a source of biochemically active substances
Blecha, Jan ; Hudeček, Jiří (advisor) ; Hodek, Petr (referee)
Sea Buckthorn (Hippophae Rhamnoides L.) as a source of biochemically active substances ABSTRACT This work is primarily focused on summarizing of existing data in the literature about the chemical composition of sea buckthorn (Hippophae rhamnoides L.) fruits, in particular of its oil. It was found that the content of individual chemical components substantially varies in different cultivars and is also very dependent on the conditions in which the plant is grown. Various sea buckthorn products can be rich sources of antioxidants, such as the vitamin C (in amounts up to 2500 mg/100g of fresh fruit) and tocopherols (up to 481 mg/100g of fresh fruit), flavonoids (up to 1100 mg/100g of fresh fruit) and carotenoids (up to 2139 mg/100g of oil). The fatty acids of this plant are mostly unsaturated (up to 89% of all fatty acids), in dominant amounts linolenic and linoleic acid can be found in seed oil and palmitooleic and saturated palmitic acid in pulp oil. Content and effects of significant flavonoids in the plant are also summarized. Among the most important biological effects of sea buckthorn, its antioxidant activity can be included as well as effects on the function of cardiovascular system, hepatoprotective and anticancer effects proven for some of the substances. Thanks to that the sea buckthorn has a high...
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NADP-dehydrogenases of medicinal plants.
Bartošová, Martina ; Hýsková, Veronika (advisor) ; Kubíčková, Božena (referee)
Medicinal plants usually contain metabolites formed by secondary biosynthetic pathways for example phenylpropanoid and shikimate pathway, which depend on NAPDH availability. In this work total content of phenolic compounds and flavonoids was determined in 10 medicinal plants common for Czech Republic. Activity of enzymes providing NADPH was determined as well. These enzymes were glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49), NADP-isocitrate dehydrogenase (NADP-ICDH, EC 1.1.1.42), NADP-malic enzyme (oxalacetate decarboxylating) (NADP-ME, EC 1.1.1.40). The activity of shikimate dehydrogenase (SDH, EC 1.1.1.25), the main enzyme of shikimate pathway, was also determined. Extraction methods were optimized for each assessment. Applicable source for NADP-dependent enzymes are black radish (Raphanus sativus L. var. niger), garlic (Allium sativum L.) and mint (Mentha piperita L). The highest content of phenolic compounds and flavonoids was determined in sage (Salvia officinalis), lemon balm (Melissa officinalis L.) and Mentha citrata. Extraction with methanol was more effective than extraction with hot water. More extraction methods and studies are necessary for characterization of individual compounds and principals on which they work. Key words: medicinal plants, NADP-dependent enzymes, phenolic...
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Complexes of zinc salts and flavonoids
Křížová, Tereza ; Roithová, Jana (advisor) ; Volný, Michael (referee)
Modern mass spectrometry (MS) is very well suited for analysis of flavonoids and their complexes. Progress during the last two decades has made MS the most sensitive method for molecular analysis of flavonoids. MS has a potential to provide information on the exact molecular mass, as well as on the structure. Flavonoids are acting as chelating agents on various metal ions. The goal of this study was to discover, which types of ions are formed in solutions of zinc salts and flavonoids by using mass spectrometry attached with electrospray ionization. First, we have studies substructures of flavonoid - resorcin, pyrokatechin a pyron - in order to find out, how individual parts of flavonoid interact with zinc ions.
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Plant alkaloids and their effects on enzymes metabolizing xenobiotics
Višněvská, Kateřina ; Stiborová, Marie (advisor) ; Černá, Věra (referee)
Sanguinarine and chelerythrine are quaternary benzo[c]phenanthridine alkaloids. The first step in sanguinarine metabolism is its reduction to dihydrosanguinarin. Antimicrobial and anti-inflammatory activities of these alkaloids are used in dentistry and as feed additives. Sanguinarine and chelerythrine induce apoptosis of cells. Fluorescence of these alkaloids and intercalation into DNA could be utilized to use the alkaloids as supravital DNA probe. Negative effect of sanguinarine and chelerythrine is their genotoxicity. Cytochrome P450 and peroxidase oxidize ellipticine to detoxication and activation metabolites. Ellipticine is a potent antineoplastic agent exhibiting the multimodal mechanism of its action. Ellipticine intercalates into DNA and inhibits topoisomerase II. Covalent DNA aducts are mediated by CYP or peroxidase oxidation of ellipticine. The anti-tumor activity of ellipticine and its derivatives is caused by a combination mechanism of cell cycle arrest and induction of the apoptotic pathway. Pharmacological efficiencies and geneotoxic side effects of ellipticine is dependent on levels and activities of cytochrome P450 or peroxidase in target tissues. Aristolactams are the major metabolites of biotransformation of aristolochic acid. Nitroreduction is the crucial step in formation of an...
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The electrochemical study of the degradation of the natural dyes from historical objects
Ramešová, Šárka ; Schwarzová, Karolina (advisor) ; Navrátil, Tomáš (referee)
The oxidation of quercetin (3,3',4',5,7-pentahydroxyflavone) and luteolin (3',4',5,7-tetrahydroxyflavone) has been studied. These intensively coloured polyphenolic compounds belong to a large group of naturally occuring flavonoid compounds. They were used with advantage as colourants in old tapestries. The study concerns the analysis of degradation processes of the original dyes. Degradation products formed during the ageing process can be used as fingerprints for the determination of the original colours. The degradation processes of flavonoid compounds were studied by electrochemical methods, absorption spectrometry and separation methods in aqueous media. The oxidation mechanism involves electron transfer coupled with chemical reaction and is influenced by the acidobasic properties of quercetin. The dissociation constants pK1 and pK2 of quercetin and pK1 and pK2 of luteolin were determined by combining of independent methods: acid-base titration and UV-Vis spectroscopy. The stability of both compounds significantly depends on the presence of the oxygen. The distribution of the oxidation products of quercetin in aqueous media within the exposure to oxygen was monitored by LC-MS/MS and LC- DAD analysis.
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Preparative diastereomeric separation of silybin and its sulfates by RP-HPLC
Kolářová, Petra ; Tesařová, Eva (advisor) ; Kalíková, Květa (referee)
Silymarin is standardized extract isolated from fruits of Milk Thistle plant (Silybum marianum). The principal component of silymarin is silybin. This flavonolignan is mainly responsible for the medicinal effects of Milk Thistle fruits: antioxidant, hepatoprotective, anticancer and chemoprotective activities. Natural silybin exists as an equimolar mixture of diastereomers A and B whose preparative separation is very hard. It was shown that the biological activity of silybin A and B are different. Silybin in the blood conjugates mainly to sulfates. The structure or biological activity of the sulfates is not yet known. The aim of this work is to develop practically applicable method for preparative separation of diastereomers of silybin A and B, and sulfates, which are considered as one of the major metabolites of silybin. The preparative method for separation of silybin A and B in the mobile phase consisting of 50% MeOH on the chromatographic column Labio C18 25x250 mm, was developed. In addition, preparative method for separation of mixture of products accured from the sulphation of silybin-23-acetate in the mobile phase consisting of MeOH/H2O 60/40 (v/v) with addition of 10 ml/l HCOOH, was optimized.
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