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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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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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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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