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Content of rutin in selected elderberry cultivars
Píšťková, Magdalena ; Hrstka, Miroslav (referee) ; Vespalcová, Milena (advisor)
The aim of this diploma thesis was to determine the rutin content in springs and leaves of 19 elderberry species (Sambucus nigra L.) with use of liquid chromatography. Furthermore the rutin content was determined in its significant sources – rue and buckwheat. The observed values were compared in order to answer the question whether it is useful to use the elderberry spring and leaves as a new source of rutin. Theoretical part of the thesis describes elderberry in general, its chemical composition and its usage in medicine. Following part concentrates on important substances contained in elderberry – flavonoids. Also principles of extraction techniques and HPLC are discussed. In practice the rutin was gained from plant matrix with use of pressurized hot water extraction (PHWE) which was performed in three five minutes long cycles. Conditions during the PHWE were set up to 80 °C and 150 bars. Obtained extract was purified with use of SPE columns and analyzed by HPLC with DAD detector (wave length – 360 nm). HPLC was performed with SUPELCOSILTMLC-8DB column. The mobile phase was composed of MeOH, H2O, and HCOOH in ration of 36:61,5:2,5. The set up flow of mobile phase was 1 ml/min. In the leaves the highest rutin content was determined in Pregarten – 26,34 ± 0,50 mg/g. On the other hand the lowest rutin content in leaves was determined in Körsör – 3,74 ± 0,13 mg/g. In springs the obtained results were as follow. The highest content was determined in Samyl – 4,87 ± 0,01 mg/g and the lowest content was determined in Heidegg 13 springs – 0,56 ± 0,01 mg/g. Also the HPLC detection limit – 6,3 . 10-5 mg/ml and quantitation limit – 2,1 . 10-4 mg/ml were determined for rutin.
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Characterisation of elderberry fruits
Christovová, Silvia ; Diviš, Pavel (referee) ; Vítová, Eva (advisor)
This master's thesis deals with the characterization of elderberry fruit (Sambucus nigra L.) with emphasis on the aroma active compounds (AAC) and vitamin C (ascorbic acid (AA), dehydroascorbic acid (DHA)). The theoretical part gives a brief outlook on the chemical composition and attributes of Sambucus berries, its healing properties and the means of processing. The rest of the theoretical part thoroughly examines the AAC and vitamin C. The suitable assessment methods are also discussed. The experimental part of the work consists of two main tasks. The first one was to assess the aroma profile of Sambucus berries using the HS-SPME-GC-FID. 18 samples were examined (the wild elderberry and 17 cultivated varieties). In total, 24 different volatile aroma active compounds were identified and quantified, 12 of which belonged to alcohols, 5 to aldehydes, 4 to esters, 1 to acids and 2 “other” compounds (linalool and -damascenone). The chemical composition of the individual samples was not uniform; no single sample contained all of the identified compounds. The second task was to assess the total vitamin C content (the sum of DHA and AA) using the HPLC. For the purpose of reducing DHA to AA three reducing agents were examined (DTT, TCEP and L-cysteine) at two different pH levels (7 and 4). TCEP at pH 4 turned out to be most efficient and was subsequently applied on the samples of 5 selected elderberry varieties. The measured vitamin C content was within the range of 5,1 – 13,0 mg per 100 g of fruit.
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Assessment of rutin in elderberries
Kaňová, Kateřina ; Hrstka, Miroslav (referee) ; Vítová, Eva (advisor)
This work is focused on determination of the concentration of rutin in the fruits of elderberry (Sambucus nigra L.). The theoretical part provides an overview of the properties, occurrence and use of elderberry in natural healing, medicine and food industry. Furthermore, important substances contained in elderberry, especially flavonoids, including rutin, are mentioned. Properties, biological effects and the possibility of determination of rutin are briefly described. In the experimental part the optimization and validation of the method of high performance liquid chromatography (HPLC), which is used for identification and quantification of rutin in fruits of 18 elderberry species, was performed. The pressurized hot water extraction was used for the preparation and solid phase extraction (SPE) for purification of the sample. The largest amount of rutin was found in a variety of elderberry Albida, 6,70 mg per 1 g of dry matter. High concentrations of rutin included varieties Haschberg, Sambu, Pregarten, Sambo and Sampo. On the other hand, the lowest concentration was found in variety Aurea, 1,24 mg.g-1. Finally, rutin content in elderberries was compared with the content of rutin in elder leaves and branches andwith significant sources of rutin – buckwheat and rue.
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Content of rutin in the biological waste material from plantations of elderberry
Boháčová, Hana ; Vránová, Dana (referee) ; Vespalcová, Milena (advisor)
This thesis is focused on determining the concentration of rutin in twigs elderberry (Sambucus nigra L.). The theoretical part provides an overview of the properties, occurrence and use of elderberry in folk medicine, medicine and food. Furthermore, there are the most important substances contained in elderberry, especially flavonoids, including rutin and quercetin as well. There are briefly described their properties, biological effects and possibilities of determination. The content of the experimental part of the thesis was developed cheap extraction methods to obtain the maximum yield of dried twigs rutin elderberry. Rutin content was observed in twigs cultivated elder varieties and wild elderberry. The identification and quantification of rutin in extracts was optimized and validated method of high performance liquid chromatography (HPLC). As a comparative method to develop extraction was used a pressurized hot water extraction (PHWE). There were tested three particle sizes of shredded twigs. The particles were extracted differently hot water varying duration. Most rutin was found in samples milled at 4 mm particles. The optimum time and temperature of extraction was 7 hours at 80°C. The highest amount of rutin in bred elderberry was determined in a sample Mladší 2014, digestion using 1,49 ± 0,004 mg.100g-1 and method PHWE 5,75 ± 0,015 mg.100g-1. There was determined the most rutin in a sample Září 2014 in the wild variant of elderberry. By digestion of ground twigs was obtained 0,90 ± 0,022 mg.100g-1 of rutin, using PHWE 2,98 ± 0,022 mg.100g-1.
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The possibility of using of elderberry fruit pomace
Ondrušová, Pavlína ; Diviš, Pavel (referee) ; Vespalcová, Milena (advisor)
This diploma thesis concerns the determination of anthocyanin pigments in the elderberry pomace (Sambucus nigra L). The theoretical part describes in general the plant specification, its chemical composition, botanical characteristics and the possibility of utilization of elderberry in food processing industry and in pharmacy. However, the main concern of this thesis are the anthocyanins. This part is closely focused on chemical structure and factors that have significant influence over their stability. There are briefly summarized the positive effects of these colourants and their use in the food industry. In the experimental part, the elderberry juice and the extracts from the rhizomes were prepared. Two different solvents, a 1: 1 water / ethanol mixture and a mixture of water and methanol in the same ratio were used as extractors. To identify and quantify the dyes in the prepared samples, a chromatographic separation system (HPLC) with a Kinetex EVO C18 column and isocratic elution of the combined mobile phases of water of acetonitrile and formic acid was developed. The analysis only took 12 minutes. Two major dyes cyanidin-3-glucoside and cyanidin-3-sambubioside were identified and quantified by HPLC. The largest amount of anthocyanins was obtained by extracting mixture methanol / water (594.6 ± 0.42 mg.100 g-1). The content of anthocyanin dyes in dry extracts and juice was also determined by the often-used pH-differential method. Both results were compared. The HPLC method provided very accurate results against the less demanding pH-differential assay method, which proved to be suitable for the indicative measurement of anthocyanin content in real samples.
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Determination of cyanide content in different parts of elderberry plant
Růžičková, Kristýna ; Štursa,, Václav (referee) ; Diviš, Pavel (advisor)
Elderberry is an abundant shrub or a small tree widely spread in the northern hemisphere. Its fruit and flowers have attracted increasing interest from consumers, pharmaceutical and food industry thanks to their high content of minerals and bioactive compounds. However, elderberry also contain potentially harmful cyanogenic glycosides which can release toxic hydrogen cyanide. The aim of this experiment was to analyse and compare total cyanide content in different plant parts of numerous interspecific hybrids, which were created at University of Maribor. Investigated plants comprised 4 subspecies of Sambucus nigra and 18 interspecific hybrids giving in total 46 elderberry genotypes. Cyanide content was determined by picrate paper method which is based on reaction of picric acid with hydrogen cyanide. The colour change of the picrate paper resulting from the reaction was further analysed using spectrophotometry. The highest cyanide levels were found in leaves reaching up to 1155.9 ± 44.8 CN- mg/kg, followed by inflorescences with maximum of 53.9 ± 3.8 CN- mg/kg, shoots and berries with highest levels of 20.3 ± 1.3 CN- mg/kg and 6.4 ± 0.0 CN- mg/kg, respectively.
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Characterization of elder juice on the basis of some chemical parameters
Mizerovská, Lucie ; Vespalcová, Milena (referee) ; Hrstka, Miroslav (advisor)
The aim of this thesis was to determine selected chemical parameters of elderberry juice namely its wild variety and four cultivated types Albida 08, Albida 09, Dana and Haschberg. The relative density was the lowest for cultivar Albida 09 (1.0149) and the highest for the wild variety of elder from the surroundings of Padochov (1.0337). Values of pH ran within the range of 3.62 for cultivar Haschberg to 4.85 for cultivar Albida 08. The highest titrating acidity was determined by cultivar Haschberg (13.12 g l-1), the lowest content of acids was determined by cultivar Albida 08 (3.47 g l-1). The highest content of reducing sugars had cultivar Albida 08 (9.64%) together with the wild variety of elder from the surroundings of Padochov (9.47 %), the minimum amount of sugars was determined by cultivar Dana (4.79 %). The content of citric acid varied between 4.76 g l-1 by cultivar Dana and 13.48 g l-1 by cultivar Haschberg. The total amino acid content, expressed as a formole number, was the lowest in cultivar Haschberg (12.9) and the highest in the wild variety of elder from the surroundings of Padochov (59.2).
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Selected alcohol extracts of Sambucus nigra for use in cosmetology
Balonková, Daniela ; Doležalová Weissmannová, Helena (referee) ; Zemanová, Jana (advisor)
This thesis focuses on the preparation of selected alcohol extracts of elderberry and their use in cosmetology. In the theoretical part are described the botanical properties of the elderberry (Sambucus nigra). There are introduced and characterized bioactive compounds and toxic compounds contained in all vegetative parts of the plant. Attention is mostly focused on compounds with antioxidant properties. The theoretical part also contains a description of basic analytical methods, which are used in the practical part to characterize compounds contained in elder flowers. The practical part focuses on the optimization of the preparation of alcohol extracts with the aim to prepare the extract with the greatest possible content of polyphenols with respect to economy and production costs. Factors such as selection of the appropriate solvent, solvent to plant material ratio, solvent temperature, and extraction time are examined. Based on the total polyphenol concentration measured spectrophotometrically by the Folin-Ciocalteu method, the most suitable solvent is 60% ethanol, the ratio of plant material to solvent is 1:8, the laboratory temperature of solvent and the extraction time of 24 hours.Under these conditions, an ethanol extract is prepared, analyzed and determined selected properties, such as refractive index and density. Rutin, chlorogenic, caffeic and ferulic acids are identified by HPLC with a DAD detector. The pH value of alcohol extract is 5.743 ± 0.006, the refractive index is 1.365 ± 0.02 at 22.7 ° C and the density is 0.9288 gcm-3. By liquid chromatography the concentration of chlorogenic acids is quantified as 0.104 gl-1, caffeic acid as 0.086 gl-1 and ferulic acid as 0.060 gl-1. The extract is incorporated into cosmetic products, that are used in balneology. Prepared products are toilet soap and bath salt.
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Determination of anthocyanins in selected kind of fruit
Forejtarová, Darina ; Vítová, Eva (referee) ; Vespalcová, Milena (advisor)
Contents of this diploma thesis was introduction of method determination of anthocyanin pigments by liquid chromatography. Anthocyanin pigments was determined in various cultivars of elderberry (Sambucus nigra L.) and in selected samples of red wine. In the first part of the theory is described elderberry, its species and chemical composition. Some possibilities of processing and utilization of elderberry in medicine are mentioned here too. The next part of the theory is aimed at chemical agents in wine, characteristics of anthocyanins and their utilization in food processing industry. The methods for determination of anthocyanin pigments with focus on HPLC are also mentioned there. The main objective of the experimental part of this thesis was comparison of content of anthocyanins in various cultivars of elderberry and in selected samples of red wine. In samples of red wine was observed not only cultivars´ influence, but also influence of the crop year on the content of anthocyanin pigments. The separation of the anthocyanin pigments by high-performance liquid chromatography with diode array detection (HPLC/DAD) was performed. The highest content of anthocyanin pigments in elderberry was determined in the cultivar Haschberg. In the case of red wine, the richest source of anthocyanins was the cultivar André 2009 – Rakvice. Higher content of anthocyanins was explicitly proved in young wines rather than in older wines.
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Evaluation of Physical and Chemical Parameters of Still Less Exploited Species of Small Fruit and Proposal of a New Soft Drink from this Fruit
Cetkovská, Jitka ; Krška,, Boris (referee) ; Barták,, Petr (referee) ; Omelková, Jiřina (advisor)
This thesis deals with still less expolited species of small fruit, evaluation of its physical and chemical parameters and proposal of a new soft drink from one of the studied species of furit. Firstly, for the characterisation of basic nutrition parameters (dry matter, content of organic acids and saccharides) and biologicaly active substances (vitamin C, anthocyanins and phenolic compounds) suitable titration, spectrophotometric, chromatographic and electrophoretic methods were chosen, optimized and validated. The sample preparation procedure was optimized as well. For three years (2010–2012) the content of listed parameters was monitored in some cultivars of five species of small fruit: in seven cultivars of sea buckthorn, in ten cultivars of cornelian cherry, in twelve cultivars of rowanberry, in two cultivars of chokeberry and in seventeen cultivars of elderberry. By means of Duncan’s test, analysis of principal components (PCA) and cluster analysis (CLU) the main differences between studied cultivars were observed mainly in sea buckthorn and rowanberry, where single cultivars were easily distinguishable from others on the basis of genetic origin. On the basis of comparison of determined parameters in studied species of fruit and with collaboration with fruit processing company the suitable species of fruit for commercial utilization was chosen – elderberry, as a important source of anthocyanins and phenolic compounds. A new soft drink as a proposal of utilization of this fruit was created – fruit juice or nectar with portion of elderberry juice. Mixed juices and nectars with various composition were prepared from elderberry and grape juice and the sensory analysis of these drinks was performed. The best evaluated were 100% fruit juices prepared with 30–50% portion of elderberry juice. The composition was jurally protected. On the basis of this protection the product consisted from 100% fruit juice with 10% of elderberry component mixed with apple and grape juice was introduced to the market by the fruit processing company.
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