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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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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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Extraction of selected elderberry flavonoids for food industry purposes
Grulichová, Hana ; Hrstka, Miroslav (referee) ; Vespalcová, Milena (advisor)
Amount of rutin and quercetin in bark and leaves from different type of Sambucus nigra was determined in this diploma thesis. Records of amounts were compared with amounts in significant sources of these flavonoids. Matters from raw matrix were obtained with pressurized solvent extraction (PSE) and pressurized hot water extraction (PHWE). There was use temperature 120°C, pressure 150 bar, three times 5-min cycles for PSE. There were used 80°C, 150 bar, three 5-min cycles for rutin extracted with PHWE and the same condition were used for extraction of quercetin only the temperature was changed to 100 °C. Analysis of rutin and quercetin were carry out high performance liquid chromatography (HPLC) with diode array detection (DAD), where was used SUPELCOSILTM LC 8DB column (5µm; 250 x 4.6 mm). Methanol: water: formic acid (36: 61.5: 2.5, pH 2.17 2.28) was used like a mobile phase. The flow of the mobile phase was set up 0.7 ml/min. Size of sample was 10 µl. The most amount of rutin was determined in the leaves of wild kind of Sambucus nigra with methanol extraction and hot water too. Extraction with PHWE was more effectively. There were determined 5.58 mg of rutin in one gram of sample from wild Sambucus nigra. There were determined the smallest amount of rutin in Sambucus nigra Körsör. There was found only 0.13 mg/g with extraction PHWE. The quercetin wasn’t determined in any Sambucus nigra leaves. There were found both of the flavonoids (rutin and quercetin) in the branch of Sambucus nigra. The rutin was determined in the small amount in wild kind but it was found in the important amount in Sambucus nigra cultivar. Whereas the quercetin was found in higher concentration than rutin when was finding the ideal conditions for extraction of branch. The quercetin was chosen like more important in light of amount. In the end the quercetin wasn’t detected in any Sambucus nigra cultivars. The most amount of quercetin was determined in wild sambucus nigra, 0.24 mg/g. The most amount of rutin was found in Sambucus nigra Albida, 2.33 mg/g.
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Possibility of determination of selected flavonoides
Smělá, Margita ; Vitoulová, Eva (referee) ; Vespalcová, Milena (advisor)
The theoretic part of the bachelor‘s thesis deals with description of selected flavonoids as apigenin, quercetin, luteolin, keampferol, rutin, myricetin and with options of their determinations. There were described typical properities and effects of several flavonoids at first. In the next part there are principles of methods, especially high-performance liquid chromatography and capillary electrophoresis. The biggest attention was made to literary survey of determination flavonoids coumpounds mentioned above, by high-performance liquid chromatography and capillary electroproresis. There were described the collection of plant material for extraction of flavonids in the practical part. The white and blue grapes were frozen by dry ice as soon as they were separated from a busch. Grapes were made in a nitrogenous atmosphere and lyophilizated after the frozen.
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Antioxidative parameters of fruit model product.
KRATOCHVÍLOVÁ, Lenka
The content of the main components (anthocyanins, quercetin, chlorogenic acid, L-ascorbic acid, rutin, total polyphenols) was determined in the syrup of elderberries (Sambucus nigra L.) by spectrophotometry or high-performanceliquid chromatography. Furthermore, the antioxidant activity of this syrup was determined using three methods: ABTS, FRAP, and DPPH. The measurement results were statistically processed and compared using correlation analysis, specifically using the Pearson correlation coefficient. It was found that the prepared product contains 2160 mg/kg anthocyanins, 201 mg/kg rutin, 26 mg/kg chlorogenic acid and 679 mg/l polyphenols. No free quercetin was found. The content of L-ascorbic acid in the original elderberries was relatively low and therefore for study purposes, the syrup was enriched with this substance in the amount of 0.5% (w/w) to study the connection of its content with other substances. It is clear from the statistical processing that the concentration of chlorogenic acid correlates with the antioxidant activity measured by the FRAP method and the concentration of polyphenols to the antioxidant activity measured by the DPPH method.
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Influence of growing conditions on the biologically active substances of non-traditional leafy vegetables.
FRIEDBERGEROVÁ, Markéta
Currently, most of the vegetables that are produced on the European continent are released in greenhouses. This method offers a considerable number of advantages, such as lower amount of water and heat consumption, protection against pests or meteorological phenomena, such as hail, and thus higher yields. However, one of the major disadvantages of these devices is the fact that glass is not permeable to UV-B radiation, which supports the biosynthesis of flavonoids, substances that have beneficial antioxidant effects for the human organism. This undesirable effect can be eliminated by installing LED (Light-Emitting Diode) lighting in greenhouses. This is because these lights provide a wide spectrum of electromagnetic radiation, i.e. wavelengths that plants use for a whole range of their processes. In this diploma thesis, the influence of growing conditions on the content of biologically active substances, especially quercetin and kaempferol in non-traditional types of Asian leafy vegetables, namely Namenia, Sagami, Golden Lion, Choy Sum and Tatsoi, was investigated. All the mentioned varieties were grown in two crops (spring and autumn) in the greenhouse and on the seedbed in the years 2019-2021. The highest concentration of total quercetin was recorded in the spring seed of Namenia grown in 2019 on the seedbed 1241 mg/kg of dry matter. Also, the highest content of total kaempferol 1753 mg/kg of dry matter was found in Namenia, it was the autumn period of 2020, also grown in a seedbed. When converted to fresh weight, the results were the same, the value for quercetin was 103 mg/kg of fresh weight and for kaempferol 142 mg/kg of fresh weight.
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Biologicky aktivní látky v netradiční listové zelenině.
FRIEDBERGEROVÁ, Markéta
Flavonoids are now increasingly coming to the forefront of interest not only from the scientific community but also of the public. They were found to be very beneficial in the field of prevention of cardiovascular or cancer diseases. In addition, they have proven to have antioxidant, anti-allergic or anti-inflammatory effects. In this bachelor 's thesis, the contents of substances, namely quercetin, kaempferol, morin, myricetin, apigenin and luteolin, were investigated using HPLC method. Lyophilized material of four varieties of Asian leafy vegetable from the cruciferous family was selected for analysis: Namenia, Sagami, Golden Lion and Tatsoi. All samples were grown in spring and autumn sowing period. Only two of the monitored aglycones, quercetin and kaempferol, were determined. The highest content of total quercetin was registered in the autumn sowing period of Namenia 1360 mg/kg of dry matter, the content of total kaempferol 1300 mg/kg of dry matter in the Golden Lion variety, also in autumn sowing period. After conversion to fresh mass, the highest concentrations of both aglycones were measured in Namenia during autumn sowing period. The amount of quercetin was determined to be 157 mg/kg of fresh weight, while the kaempferol content was 114 mg/kg of fresh weight.
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Enzymatic and Metabolic Transformation of Silybin and its Congeners
Purchartová, Kateřina
Natural flavonoids and flavonolignans feature beneficial properties for living organisms such as antioxidant and hepatoprotective effects, anticancer, chemoprotective, dermatoprotective and hypocholesterolemic activities. Their metabolism in mammals is complex, the exact structure of their metabolites still remains partly unclear and the standards are usually not commercially available. Hence, this project focused on the preparation of potential and defined biotransformation Phase II sulfated metabolites of silymarin flavonolignans: silybin, 2,3-dehydrosilybin, isosilybin, silychristin, silydianin and flavonoids quercetin, taxifolin, rutin and isoquercitrin. Pure sulfated derivatives were prepared using aryl sulfotransferase from Desulfitobacterium hafniense and aryl sulfotransferase from rat liver. Using heterologously expressed PAPS (3'-phosphoadenosine-5'-phosophosulfate) - independent arylsulfotransferase from Desulfitobacterium hafniense and cheap p-nitrophenyl sulfate as sulfate donor, sulfated flavonolignans and flavonoids were obtained in high yields. Silymarin flavonolignans afforded exclusively monosulfates at the position C-20 (C-19 in the case of silychristin), except 2,3-dehydrosilybin that yielded also the 7,20-O-disulfated derivative. Isoquercitrin and rutin were selectively sulfated...
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Monitoring of biologically active substances in Asian vegetables from Brassicaceae family
SVOBODOVÁ, Martina
The Bachelor thesis is monitoring the content of polyphenolic substances in non-traditional leafy vegetables at different times of the year. Specific samples were experimentally studied, such as: green mustard (Brassica rapa var. komatsuna), red mustard (Brassica juncea (L.) Czern.), mizuna and mibuna (Brassica rapa ssp. Japonica (L.)). Although the samples are Asian leafy vegetables, they are now widely used in the Czech Republic and can be found both in the form of seeds and as leaves for consumption. Therefore, the issue of beneficial biologically active substances is topical. The phenolic group includes flavonoids that are easily available and their biological activity is also significant. Vegetables have antioxidant properties due to flavonoids, which prevent lipid peroxidation and eliminate free radicals. This property is also beneficial to humans, and therefore flavonoid substances are also used in medicine. They can prevent the emergence of chronic diseases such as cardiovascular disease or atherosclerosis. Flavonoids are also attributed to preventive effects against cancer and diabetes mellitus. Samples of cruciferous leafy vegetables were grown in 2017 on the experimental plot of the Faculty of Agriculture of the University of South Bohemia in České Budějovice on the open area of the flowerbed in spring and autumn sowing. The high performance liquid chromatography method was used to measure the content of flavonoid substances. The following substances were studied in selected vegetables: myricetin, morin, luteolin, quercetin, apigenin and kaempferol. Unfortunately, myricetin, morin, luteolin and apigenin were not detected by HPLC. Concentrations of only quercetin and kaempferol were measured for final comparison with expert articles. The sowing time did not affect the measured contents quercetin and kaempferol in most of the samples. Only a specimen of Chinese red mustard from autumn sowing contained more biologically active substances.
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Biologically active phenolic compounds in small fruit.
LAXOVÁ, Lenka
This master´s thesis focuses at bioactive compounds in edible berries during its culinary processing. Elderberries (Sambucus nigra L.) were selected as a representative of edible berries variety. Elderberry is known for presence of significant bioactive compound content and usage in traditional medicine. Elderberries were processed acording to common culinary recipes. Amount od selected bioactive polyphenolic compounds (chlorogenic acid, quercetin, rutin, anthocyanes) was detected accordingly. Additionally content of ascorbic acid was determined. On the basis of the experimental determination, it was found that anthocyanes and rutin are the most common compounds found in elderberries. All analytes in juices and sirups gradually decreased during the treatments and long-term storage. On the contrary, the content of quercetin increased with gradual modifications and storage. The highest decrease was observed for vitamin C content. The lowest values in the products from elderberries were reached by quercetin and ascorbic acid.
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