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Isolation of active substances with antioxidative properties from lignocellulosic waste
Jurová, Karolína ; Vítová, Eva (referee) ; Kovalčík, Adriána (advisor)
This thesis focuses on isolating active substances with antioxidant properties from lignocellulosic wastes. This work aimed to modify bacterial cellulose (BC) with extracts from grape pomace, which show high antioxidant activity. Grape pomace belongs to the group of lignocellulosic wastes with high content of bioactive compounds such as polyphenols, which contribute significantly to antioxidant efficacy. This work tested the hypothesis that grape extracts can provide antioxidative efficiency to BC. Grape pomace extracts were prepared from grape pomace frozen and lyophilized using a 70% acetone, a 70% ethanol or a 50% ethanol solvent. The highest values of antioxidant activity were determined for the lyophilized extract prepared by extracting grape pomace with 70% acetone. The extracts were further used to modify bacterial cellulose in the form of films (dry BC) and hydrogels (wet BC). It was shown that both forms of BC could be effectively modified using grape pomace extracts, but BC hydrogel achieved higher antioxidant activity compared to BC film. Furthermore, BC with antioxidant activity can be used in various food, cosmetics, and medicine applications.
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Influence of production environment on the occurrence of contaminating Penicillium in smear ripened cheese
Veselá, Vendula ; Brázda, Václav (referee) ; Vítová, Eva (advisor)
The topic of this thesis is the monitoring of the environment in a cheese production plant specializing in cheese ripened under a smear rind. The aim is to evaluate the occurrence of Penicillium mold as the main contaminant microorganism, depending on the sampling location and production time. Air samples were collected throughout the production process using the sedimentation method at 21 designated locations. The total number of microorganisms, selected groups of microorganisms, and the target microorganism (Penicillium mold) were monitored in the collected samples using cultivation (according to ČSN standards), microscopic, and molecular diagnostic techniques. The aim was to evaluate the occurrence of Penicillium citrinum, chrysogenum and commune species in dependence on the sampling location and production time. As part of the production screening, the broader microflora was first evaluated, followed by monitoring for contaminating molds. Using macroscopic and microscopic methods, 36 morphologically distinct colonies were selected. Microscopic analysis confirmed the presence of bacteria, yeasts and molds of the genera Penicillium, Cladosporium and Aspergillus. In order to identify the microorganisms present, an analysis using the polymerase chain reaction (PCR) method was performed, preceded by DNA isolation using the NucleoSpin Microbial DNA kit and phenol extraction. Based on real-time PCR results, the occurrence of contaminating Penicillium and Cladosporium molds was detected even in the early stages of production. Molds from these genera are found throughout the entire facility except for the single room that serves as a curd cooler before forming. Aspergillus mold, in the form of yellow mold with white mycelium on the edge of the colony, was detected only once in the drying room where cheese ripening occurs, and confirmed by microscopic observation. The genus Cladosporium was found in five morphologically distinct forms in the production environment. In addition, DNA was isolated from 11 morphologically distinct bacteria. Real-time PCR identified this DNA as belonging to coryneforms, with four of them belonging to the genus Brevibacterium. The genus Penicillium was represented by 13 morphologically distinct colonies, real-time PCR confirmed the presence of Penicillium commune and Penicillium chrysogenum. In addition to the mentioned molds, 11 morphologically distinct bacteria were detected, from which DNA was isolated. Real-time PCR identified this DNA as belonging to coryneforms, with 4 of them belonging to the genus Brevibacterium. The results of this work completely map the contamination of the production environment of the plant, and based on these results, methods for environmental sanitation were proposed to ensure the decontamination of the production process.
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Developement and characterization of high protein plant-based yogurt
Smatana, Igor ; Vítová, Eva (referee) ; Pořízka, Jaromír (advisor)
The thesis was focused on the production and characterization of plant-based yogurts with increased protein content. The aim was to design and optimize the formula of new plant-based products. The motivation for this work was the ever-increasing demand for plant-based food alternatives in recent years. The theoretical part was focused on the trend of development of dairy alternatives as well as the nutritional challenges associated with modern diets such as vegetarianism and veganism. Focus was also on the nutritional importance of protein and the composition of proteins of plant origin. The impact of live bacteria on human health, as well as common analytical methods for determining rheological and sensory properties was also included. In the experimental part, the methods used for the preparation and characterization of novel plant-based high-protein yogurts were described. The results showed that the biggest challenge in the production of plant-based yogurts, without added stabilizing agents, is to achieve an acceptable texture, so that the products are neither too watery nor too tough, while maintaining suitable sensory properties. For the products with the addition of 20 g of almond protein, this texture was almost achieved, while maintaining the appropriate sensory characteristics. In contrast, for products containing 10 g almond protein and 6 g of pea or rice protein, the texture was unacceptable and the products were splitting into 2 phases.
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Jelly candies with the addition of bilberry (Vaccinium myrtillus) extract
Knapovská, Pavlína ; Pořízka, Jaromír (referee) ; Vítová, Eva (advisor)
This thesis deals with the application of bilberry fruits (Vaccinium myrtillus) and their extract to jelly candies and the subsequent characterization of these candies in terms of volatile compounds, total phenolic compounds and anthocyanins, antioxidant activity and sensory quality. The volatiles were determined by headspace solid-phase microextraction coupled to gas chromatography with mass spectrometry and standard spectrophotometric methods were used for the determination of bioactive compounds. Total phenolic content was determined by the Folin-Ciocalteu method, antioxidant activity by the TEAC method and total anthocyanin content by pH differential analysis. The sensory quality of the samples was evaluated using unstructured graphical scales. The preparation of the bilberry extract was optimized on the basis of total phenolic content and the following conditions were selected as the optimal: a weight of 1000 grams of bilberries per litre of water, a temperature of 50 °C, and an extraction time of 2 hours. The optimum additions to candies were selected, by sensory analysis, as 25 wt. % of extract and 15 wt. % of bilberries. A total of 45 volatile compounds were identified in the candy with bilberry extract, the most abundant being (E)-9-hexadecen-1-ol (22,7 %), furfural (10,9 %) and 2-ethylhexan-1-ol (10,0 %). The total phenolic content was 1141 ± 11 gGAE · g-1, the antioxidant activity was 660 ± 66 gTE · g-1 and the total anthocyanin content was 418 ± 23 gC3G · g-1. A total of 40 volatile compounds were identified in the candy with bilberry and the most abundant were linalool (22,7 %), (E)-9-hexadecene-1-ol (16,1 %) and 2-ethylhexan-1-ol (8,4 %). The total phenolic content was 1735 ± 56 gGAE · g-1, the antioxidant activity was 1109 ± 32 gTE · g-1 and the total anthocyanin content was 745 ± 23 gC3G · g-1. With the addition of both the extract and the bilberries, the candies showed a significant increase in bioactive and volatile compounds content, and the colour and flavour of the enriched samples were positively evaluated, thus improving their overall acceptability.
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Determination of selected bioactive substances in grapes
Havlíková, Markéta ; Trachtová, Štěpánka (referee) ; Vítová, Eva (advisor)
The subject of this thesis was the determination of selected bioactive substances in grapes and the subsequent comparison of differences between the samples. Two fractions of grapes (skins and pulp with seeds) of 4 grape varieties - Hibernal, Johanniter, Riesling and Aurelius - were analysed. Two methods were used for the determination: for phenolic substances, high-performance liquid chromatography with mass spectrometry and for volatile substances, headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry. For each of these methods, the optimal extraction technique was first selected and then used for the analysis of all samples. The linearity of the external standard method was also verified with a reliability coefficient R2 in the range of 0.9650-0.9989 for high-performance liquid chromatography with mass spectrometry and 0.8571-0.9999 for headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry. 63 volatile and 9 phenolic compounds were identified in the samples. From these, specific compounds of grapes (20 volatile compounds and 5 phenolic compounds) were selected. On the basis of the amount of observed compounds, the two grape fractions and subsequently the grape varieties were compared with each other. There were differences in the content of selected compounds between grape fractions and across varieties, which could probably be due to the geographical origin of the samples.
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Classic vs. bulgarian yogurt - sensory quality vs. chemical and microbiological composition
Podloučková, Michaela ; Němcová, Andrea (referee) ; Vítová, Eva (advisor)
This thesis compares classic and Bulgarian yoghurt in terms of volatile content, sensory quality, and microbial profile. The volatiles were identified and semi-quantified by gas chromatography with mass detection in conjunction with solid phase microextraction, whereas classical culture followed by real-time polymerase chain reaction (PCR) was used to determine the microbial profile. Sensory analysis consisted of evaluation by graphic scales, profile and ordinal tests. In the experimental part, 3 samples were produced, a "classic" white and Bulgarian yoghurt made from commercial yoghurt cultures, and a traditional Bulgarian yoghurt imported directly from Bulgaria. Based on the PCR analysis performed, bacterial DNA was confirmed in all yogurts, only the traditional Bulgarian yogurt also contained yeast DNA. The Bulgarian traditional yoghurt thus differed in its detectable yeast taste, but above all in the qualitative and quantitative composition of the volatile substances. Of the 29 compounds identified, 20 were present in this yoghurt; esters were the main group, while ketones predominated in the other yoghurts. From a sensory point of view, the Bulgarian yoghurt made from commercial culture was the best evaluated, with a pleasant appearance, a pleasant smell and a pleasantly sour taste and an overall excellent sensory quality.
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Study of the effect of adding coffee grounds to the soil on the growth of selected plants
Kopková, Pavlína ; Vítová, Eva (referee) ; Diviš, Pavel (advisor)
Due to the globally high consumption of coffee, a large amount of spent coffee grounds is generated. This study aims to evaluate the potential use of this waste material in agriculture as fertiliser for field crops. Growth experiments were conducted with brown soil enriched with 2.5 vol.% of coffee grounds or modified coffee grounds, followed by soil and lettuce (Lactuca sativa L.) and corn (Zea mays) plant characterisation. Although germination tests did not demonstrate phytotoxicity of the soil mixtures, the addition of any modified coffee grounds led to a decrease in plant biomass. Differences between lettuce and corn were observed during growth. After harvesting, a decreased chlorophyll content or mineral substances was found in the cultivated plants. Prepared soil mixtures affected soil pH, with the greatest pH decrease measured after the addition of oxidized coffee grounds. Elemental analysis of the soils before cultivation showed an increase in the content of some essential mineral elements for plants (K, Ca, Na, Mg), but at the same time, there was a decrease in the concentration of one of the most important elements, phosphorus (P).
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Optimizing the production of non-chocolate sweets with the addition of Filipendula ulmaria extract using various sweeteners
Malyszová, Markéta ; Diviš, Pavel (referee) ; Vítová, Eva (advisor)
The aim of this master's thesis was to optimize the production process of jelly candies using various alternative sweeteners and subsequently adding herbal extract of meadowsweet, achieving not only better nutritional properties but also enhanced sensory characteristics of the final product. Sensory analysis was utilized to determine the ideal combination of sweetener type and amount, which most closely aligns with consumer preferences for sweetness and texture, while maintaining a low energy component content. Based on these criteria, maltitol was selected as the sweetener in a 50% w/w solution. The study also focused on optimizing the addition of Filipendula ulmaria extract to the candies made with the "ideal" sweetener. Again, using sensory analysis, the optimal concentration of the extract and the type of gelling agent were selected to achieve the desired consistency, taste, and other sensory parameters that make the product attractive for consumers. Following these experiments, the ideal values were determined to be 50% w/w for sweetener, a 5 % addition of Filipendula ulmaria extract, and the use of gelatin as the gelling agent. Subsequently, a chemical analysis of the produced samples and meadowsweet extract was also carried out. Identification and quantification of volatile aromatic compounds were performed using the HS SPME GC-MS method, which identified 39 different substances, predominantly from the group of alcohols and aldehydes. To determine the total content of phenolic compounds, the Folin-Ciocalteu spectrophotometric method was used, and the pH differential method was employed to determine the content of anthocyanins. The total content of phenolic compounds in the extract amounted to 1418.16 ± 6.56 gml-1, and the total anthocyanin content was determined to be 249.19 ± 27.40 gml-1. Antioxidant activity was tested using two methods, TEAC and DPPH. The results of the antioxidant activity measured by the DPPH method were higher, confirming the greater robustness of this method. However, both tests confirmed the antioxidant activity of the tested samples. Finally, the antimicrobial activity of the samples was tested using the diffusion well method against strains of Bacillus cereus and Escherichia coli. The samples exhibited antimicrobial activity against both tested microorganisms, but showed more effective action against Bacillus cereus, which may be due to the simpler structure of their cell wall.
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Assessment of fatty acids in selected type of cheese
Chmelařová, Adéla ; Sůkalová, Kateřina (referee) ; Vítová, Eva (advisor)
This bachelor thesis is focused on identification and quantification of free and bound fatty acids in commercially available white mold cheeses: Hermelín, Kamadet and Olmín, using gass chromatography withflame ionization detection. Theoretical part of the thesis is dealing with the classification of cheeses with the main attention to white mold cheeses. In a following part cheese production technology is described, lipids and fatty acids are characterised including the possible methods for their assessment, mainly gass chromatography. Experimental part the contents of free and bound fatty acids in selected samples of white mould cheeses were compared. The method in accordance with ČSN EN ISO 1735:2005 was selected for lipid extraction. Fatty acids were identified as methylesters using acid esterification with methanolic solution of bortrifluorid as a catalyst. In total 21 free and bound fatty acids were detected in samples.
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Characterisation and maturation process of Edam type cheese
Dostálková, Andrea ; Vespalcová, Milena (referee) ; Vítová, Eva (advisor)
This diploma thesis deals with the assessment of aroma compounds and free/bound fatty acids in model samples of Edam type cheeses. Various combinations of microbial cultures, basic mesophilic and four selected strains of thermophilic Lactobacillus casei and Lactobacillus plantarum, were used for cheese production. The main aim was to judge the influence of microbial cultures on content of measured parameters, the changes during ripening were also followed. The HS-SPME-GC-MS method was used for identification and semiquantification of aroma compounds. The lipids from cheese were extracted based on ISO 1735:2004, fatty acids were identified and quantified using GC-FID method after conversion to methyl esters by acid esterification with boron trifluoride in methanol. In total 50 volatile compounds were identified in samples, 11 alcohols, 5 aldehydes, 7 ketones, 6 acids, 10 esters, 3 lactones, 2 sulphur compounds and 6 „others“; all of them are considered as aroma active. About 30–40 compounds were identified in single samples, alcohols, acids and ketones were the predominant. The number as well as the content of compounds identified decreased slightly during ripening. In total 17 free/bound fatty acids were identified in samples, their content also decreased during ripening. Myristic, palmitic, stearic and oleic acids were the predominant. The significant differences (p
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