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Determination of athenticity of plant foods by molecular techniques
Plášková, Anna ; Mikulíková, Renata (referee) ; Němcová, Andrea (advisor)
The aim of presented diploma thesis was to determination of authenticity of fruit baby foods for early infant feeding using molecular methods. In the experimental part, isolation kit was used for isolation of plant DNA from fruits (strawberry, apricot, raspberry, apple) and from six commercial fruit products for children. Isolated DNA was characterized and verified using PCR methods with primers specific for plant rDNA (ITS2). Specific primer pairs were designed to amplify DNA for the detection of one fruit species. Primer specificity was assessed with four fruit species. A mixture of fruit puree from the two fruits was used to determine the sensitivity of the multiplex PCR assay. Six commercial fruit products were evaluated to verify the applicability of the multiplex PCR assay. The methodology of molecular detection of fruit DNA by qPCR and multiplex qPCR (duplex) includes approaches, which enable to detect two fruits (strawberry-raspberry, apricot-apple) in one reaction and thus reduces time and money requirements.
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Detection of selected fruit species in plant-based foodstuffs through instrumental analysis and methods of molecular biology
Ondruch, Petr ; Langová, Denisa (referee) ; Fialová, Lenka (advisor)
Food is often subject to adulteration, including fruit products containing strawberries. This work deals with the detection of strawberries in model and real fruit products, so that possible falsification can be detected. The aim of the work was the preparation of model and real fruit products, their analysis using selected instrumental and molecular-biological methods and mutual comparison of the results of these methods. Model mixtures of fruit products were prepared and commercial ones were purchased. The content of polyphenols and flavonoids was determined by UV-VIS spectrophotometry, which were characterized in more detail by HPLC. DNA was isolated from the fruit purees, suitable primers confirmed the amplifiability and formation of a specific product for strawberry, and this product was verified by electrophoretic control. In the instrumental part, certain substances were detected by extraction from model and commercial mixtures and the HPLC method, and the amount of selected substances was determined. The lowest amount of strawberry DNA that could be detected by selected primers in the PCR method was 0.7 ng /µl, in commercial mixtures this DNA could not be detected. In the HPLC method, the strawberry was detected in all samples by focusing on a particular fingerprint and the amount of pelargonidin-3-glucoside in the samples. The instrumental method can be considered more suitable in this case.
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Determination of athenticity of plant foods by molecular techniques
Plášková, Anna ; Mikulíková, Renata (referee) ; Němcová, Andrea (advisor)
The aim of presented diploma thesis was to determination of authenticity of fruit baby foods for early infant feeding using molecular methods. In the experimental part, isolation kit was used for isolation of plant DNA from fruits (strawberry, apricot, raspberry, apple) and from six commercial fruit products for children. Isolated DNA was characterized and verified using PCR methods with primers specific for plant rDNA (ITS2). Specific primer pairs were designed to amplify DNA for the detection of one fruit species. Primer specificity was assessed with four fruit species. A mixture of fruit puree from the two fruits was used to determine the sensitivity of the multiplex PCR assay. Six commercial fruit products were evaluated to verify the applicability of the multiplex PCR assay. The methodology of molecular detection of fruit DNA by qPCR and multiplex qPCR (duplex) includes approaches, which enable to detect two fruits (strawberry-raspberry, apricot-apple) in one reaction and thus reduces time and money requirements.
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|
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Detection of selected fruit species in plant-based foodstuffs through instrumental analysis and methods of molecular biology
Ondruch, Petr ; Langová, Denisa (referee) ; Fialová, Lenka (advisor)
Food is often subject to adulteration, including fruit products containing strawberries. This work deals with the detection of strawberries in model and real fruit products, so that possible falsification can be detected. The aim of the work was the preparation of model and real fruit products, their analysis using selected instrumental and molecular-biological methods and mutual comparison of the results of these methods. Model mixtures of fruit products were prepared and commercial ones were purchased. The content of polyphenols and flavonoids was determined by UV-VIS spectrophotometry, which were characterized in more detail by HPLC. DNA was isolated from the fruit purees, suitable primers confirmed the amplifiability and formation of a specific product for strawberry, and this product was verified by electrophoretic control. In the instrumental part, certain substances were detected by extraction from model and commercial mixtures and the HPLC method, and the amount of selected substances was determined. The lowest amount of strawberry DNA that could be detected by selected primers in the PCR method was 0.7 ng /µl, in commercial mixtures this DNA could not be detected. In the HPLC method, the strawberry was detected in all samples by focusing on a particular fingerprint and the amount of pelargonidin-3-glucoside in the samples. The instrumental method can be considered more suitable in this case.
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