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Testing of primers for real-time PCR-HRM analysis of fruit products containing one fruit species
Boháčová, Barbora ; Dzurendová, Simona (referee) ; Fialová, Lenka (advisor)
Determining the authenticity of fruit products, which are often counterfeited by substituting part of a more expensive fruit with a cheaper but botanically similar fruit, is a current topic in the food industry. A prime example is the dilution of apricot products with peach puree. This study focuses on testing specific primers AGS18 and PdCass to reveal the true proportion of apricot content in model products mixed with peach puree using PCR analysis followed by HRM analysis. Testing of these primers for authenticity determination revealed limited utility of AGS18 primers due to the formation of small amount or no specific products during reactions with fruit DNA. PdCass primers required PCR condition optimization, but subsequently, specific DNA sequences from fruit leaves could be amplified in sufficient quantities. PCR analysis of DNA from model products with PdCass primers provided specific products in samples with apricot content of 70% or less. HRM analysis of samples and calculation of GCP did not distinguish purees with 0%, 10%, and 30% apricot content. However, purees with 50% and 70% apricot content exhibited significantly different melting curves compared to other samples.
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Phloridzin content in selected apple varieties
Křehlíková, Jana ; Mikulíková, Renata (referee) ; Diviš, Pavel (advisor)
Phloridzin, a phenolic compound found in Malus genus, was targeted in this study for detection in apples and apple-based foods via HPLC. The research aimed to assess phlorizin’s potential as a marker for confirming apple presence in food, addressing food authenticity concerns. The methodology encompassed method optimization for sample preparation, considering solvent selection and oxygen impact on stability. Analysis revealed significant phloridzin distribution among apple varieties (Golden Delicious, Red Delicious, Granny Smith, Gala, Champion, Jonagold), averaging 17 ± 10 mgkg1. Further, 15 fruit products were surveyed for phloridzin content, with 5 non-apple-labelled foods also analysed. Phloridzin proved effective in detecting undeclared apple presence in jams, fillings, and fruit purees, highlighting potential for authenticity verification. However, fruit drinks and juices exhibited inconclusive results, suggesting a need for improved sample preparation and analysis methods.
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Authenticity of natural plant component in cosmetics products
Kubalová, Michaela ; Fialová, Lenka (referee) ; Němcová, Andrea (advisor)
The purpose of this thesis was to study the authenticity of selected natural ingredients in cosmetic products. These were specifically cosmetic products that contained citruses, mint or lavender. Commercially available isolation kits were used for DNA isolation. The presence of plant origin DNA was verified by PCR method using primers specific for the ITS2 region of plants. The presence of limonene, a significant allergen contained in said plants, was determined in the samples by PCR method using primers for limonene synthase. At the same time, its presence was verified by HPLC method. In addition, two primers were tested for lavender and monitored for their efficacy, with no significant difference in the usage.
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Study of geographical authenticity of Moravian white wines
Horáková, Kristýna ; Křikala, Jakub (referee) ; Diviš, Pavel (advisor)
The aim of this bachelor thesis was to verify geographical autenticity of Moravian white wines based on elemental analysis. This data were completed with the analysis of organic acids, fenolic compounds and atioxidants in wines. There were used two spektrometric methods for the elemental analysis (ICP-MS and ICP-OES). The concentration of organic acids was mesured by Ion chromatography. To determine the concentration of phenolic compounds in wines there was used the high performance liquid chromatography (HPLC-DAD), the content of antioxidants and the total content of phenols was measured on spectrophotometer. In total there were 33 samples of wine from 4 Moravian subregions. For the purpose of determination the samples in groups the dataset was firstly corrected by Grubbs test for remote results. From those data the box plots were created which represents the minimal, maximal and mean concentrations of each measured component. Furthermore the analysis of variance was applied, which picked up 12 statisticly valuable parameters from the whole 47. By appliyng the multidimensional statistical methods (Pearson correlation analysis (PCA) and discriminant analysis (DA)) were compiled mathematical models for the division of Moravian wines according to geographical origin. The success rate of the discrimination test was 96,97%. The cross-validation test showed an 81,82% success rate for classifying unknown samples.
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Development of methods for determination of authenticity of cosmetic products with a plant component
Langová, Denisa ; Brázda, Václav (referee) ; Vorlová, Lenka (referee) ; Márová, Ivana (advisor)
In the final thesis, methods were developed for determining the authenticity of cosmetic products and food products with a plant component. The presence of natural ingredients in a series of model and commercial cosmetic products was verified and their authenticity was verified, too. The focus of the work was the development of a molecular-biological method for the detection of plant components (herbs, fruits, essential oils) and probiotics present in cosmetic products. Commercially available isolation kits were used for the isolation of DNA from cosmetic products and the method was optimized for the isolation of plant DNA present in cosmetic products. The presence of a plant matrix in cosmetic products was detected by the polymerase chain reaction (PCR) method. Instrumental methods were used to detect the presence of fragrance allergens in cosmetic products. A method of quantifying these allergens using liquid chromatography was introduced and the measurement results were compared with the results of gas chromatography with mass detection. The proposed method of isolation and subsequent amplification of plant DNA is suitable for determining the authenticity of herbal ingredients in cosmetic products. The proposed HPLC method is a simple and inexpensive method suitable for determining the allergens present in the amount necessary to comply with EU analyte legislation and in the required sensitivity (except for hydroxycitronellal). Both methods are suitable to be used for the analysis of food products, too.
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Authenticity of natural plant component in cosmetics products
Kubalová, Michaela ; Fialová, Lenka (referee) ; Němcová, Andrea (advisor)
The purpose of this thesis was to study the authenticity of selected natural ingredients in cosmetic products. These were specifically cosmetic products that contained citruses, mint or lavender. Commercially available isolation kits were used for DNA isolation. The presence of plant origin DNA was verified by PCR method using primers specific for the ITS2 region of plants. The presence of limonene, a significant allergen contained in said plants, was determined in the samples by PCR method using primers for limonene synthase. At the same time, its presence was verified by HPLC method. In addition, two primers were tested for lavender and monitored for their efficacy, with no significant difference in the usage.
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Study of geographical authenticity of Moravian white wines
Horáková, Kristýna ; Křikala, Jakub (referee) ; Diviš, Pavel (advisor)
The aim of this bachelor thesis was to verify geographical autenticity of Moravian white wines based on elemental analysis. This data were completed with the analysis of organic acids, fenolic compounds and atioxidants in wines. There were used two spektrometric methods for the elemental analysis (ICP-MS and ICP-OES). The concentration of organic acids was mesured by Ion chromatography. To determine the concentration of phenolic compounds in wines there was used the high performance liquid chromatography (HPLC-DAD), the content of antioxidants and the total content of phenols was measured on spectrophotometer. In total there were 33 samples of wine from 4 Moravian subregions. For the purpose of determination the samples in groups the dataset was firstly corrected by Grubbs test for remote results. From those data the box plots were created which represents the minimal, maximal and mean concentrations of each measured component. Furthermore the analysis of variance was applied, which picked up 12 statisticly valuable parameters from the whole 47. By appliyng the multidimensional statistical methods (Pearson correlation analysis (PCA) and discriminant analysis (DA)) were compiled mathematical models for the division of Moravian wines according to geographical origin. The success rate of the discrimination test was 96,97%. The cross-validation test showed an 81,82% success rate for classifying unknown samples.
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Where are the Boundaries of Normality? The Search for Autistic Subjectivity
Polčová, Lenka ; Stöckelová, Tereza (advisor) ; Szénássy, Edit (referee)
Despite of the growing body of multidisciplinary literature dealing with autism, encounters with a reality on the spectrum remains an under researched area. During my two years long fieldwork I discovered the controversial nature of being on the spectrum. To be on the spectrum is at the same time similar and different from the kind of situations in which any human being is entangled. This diploma thesis follows the process of formation of being on the spectrum situated in specific "playful" environment. It focuses on heterogeneous interactions among human and nonhuman agents. It follows multiple (a)symmetries that emerge in this process. The aim of the thesis is to introduce the reality on the spectrum in an innovative and processually situated way and making it a leverage of exploring the nature of the social. The ethnographic research took place in four "playrooms" and mapped the material practices of the alternative therapy Son-Rise. Through mirroring, the main therapeutic practice, boundaries of similarity/otherness where the other is emerging as an autist subject are continuously negotiated, generated and blurred. Becoming similar comprises of multiple practices, including assuming other (autistic) sensitivity and inventing similar (body) competencies. In addition, the diploma thesis discusses...
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