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Preparation of food supplement containing probiotic bacteria and components with anti-inflammatory effect
Horňáková, Nikola ; Němcová, Andrea (oponent) ; Skoumalová, Petra (vedoucí práce)
The presented thesis discusses the issues of chronic inflammatory diseases of the digestive system and suggests the possibility of the alternative natural remedies improving the health conditions or prolonging the remission phase of IBD. The main goal is to design a probiotic supplement enriched by natural anti-inflammatory agents. Phytochemicals, concretely phenolic compounds, flavonoids, and carotenoids suppress harmful inflammatory processes by direct targeting the function of the immune cells or by inhibiting damaging oxidative stress in general. Therefore, there were several plants potentially rich for these biologically active substances selected. Concretely, blueberry (Vaccinium myrtillus), lingonberry (Vaccinium vitis-idaea), turmeric (Curcuma longa), peppermint (Mentha piperita), chamomile (Matricaria recutita), cinnamon (Cinnamomum zeylanicum), ginger (Zingiber officinale) and flax (Linum usitatissimum). The suitable parts of these plants were extracted by using a total of three different solvents – water, ethanol, and hexane for obtaining lipidic extracts. The concentration of total polyphenols, total flavonoids, total carotenoids, and the ascertainment of specific polyphenols was determined in the prepared extracts. These characteristics were enhanced by the measurement of antioxidant activity of the aqueous and ethanolic extracts. The interaction of individual samples with probiotic cultures (Bifidobacterium breve, Lactobacillus acidophilus) and the ability of extracts to influence the viability of probiotics in the model digestion process has been examined. The most interesting samples (aqueous extracts of blueberries, mint, chamomile, and cinnamon) were selected for further experiments. The possible cytotoxicity towards human intestinal epithelial cells was tested in vitro by the MTT tests utilizing the CaCo-2 cell line. Extracts showing the highest levels of beneficial phytochemicals and antioxidant activity, supporting the growth of probiotic cultures, and showing minimal cytotoxic effects on human intestinal cells were then co-encapsulated with probiotics into alginate particles of a diameter of 1 mm. Water extracts of mint, chamomile, and cinnamon have been chosen. The encapsulation efficiency of successfully entrapped probiotics and phenolic compounds was determined in prepared particles. Furthermore, the particles were studied during the process of model digestion, when the release of the desired substances in the various parts of the digestive system was observed and assessed whether the components would reach the crucial point of action – the colon. For the use of the proposed probiotic mixture as a dietary supplement, a recommended dose of 1 g has been determined. Lastly, possible adjustments such as lyophilization or sheathing by an extra protective polymerous layer, e.g. chitosan, were suggested to prolong the shelf life of the particles and volatile substances stability.
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Preparation of food supplement containing probiotic bacteria and components with anti-inflammatory effect
Horňáková, Nikola ; Němcová, Andrea (oponent) ; Skoumalová, Petra (vedoucí práce)
The presented thesis discusses the issues of chronic inflammatory diseases of the digestive system and suggests the possibility of the alternative natural remedies improving the health conditions or prolonging the remission phase of IBD. The main goal is to design a probiotic supplement enriched by natural anti-inflammatory agents. Phytochemicals, concretely phenolic compounds, flavonoids, and carotenoids suppress harmful inflammatory processes by direct targeting the function of the immune cells or by inhibiting damaging oxidative stress in general. Therefore, there were several plants potentially rich for these biologically active substances selected. Concretely, blueberry (Vaccinium myrtillus), lingonberry (Vaccinium vitis-idaea), turmeric (Curcuma longa), peppermint (Mentha piperita), chamomile (Matricaria recutita), cinnamon (Cinnamomum zeylanicum), ginger (Zingiber officinale) and flax (Linum usitatissimum). The suitable parts of these plants were extracted by using a total of three different solvents – water, ethanol, and hexane for obtaining lipidic extracts. The concentration of total polyphenols, total flavonoids, total carotenoids, and the ascertainment of specific polyphenols was determined in the prepared extracts. These characteristics were enhanced by the measurement of antioxidant activity of the aqueous and ethanolic extracts. The interaction of individual samples with probiotic cultures (Bifidobacterium breve, Lactobacillus acidophilus) and the ability of extracts to influence the viability of probiotics in the model digestion process has been examined. The most interesting samples (aqueous extracts of blueberries, mint, chamomile, and cinnamon) were selected for further experiments. The possible cytotoxicity towards human intestinal epithelial cells was tested in vitro by the MTT tests utilizing the CaCo-2 cell line. Extracts showing the highest levels of beneficial phytochemicals and antioxidant activity, supporting the growth of probiotic cultures, and showing minimal cytotoxic effects on human intestinal cells were then co-encapsulated with probiotics into alginate particles of a diameter of 1 mm. Water extracts of mint, chamomile, and cinnamon have been chosen. The encapsulation efficiency of successfully entrapped probiotics and phenolic compounds was determined in prepared particles. Furthermore, the particles were studied during the process of model digestion, when the release of the desired substances in the various parts of the digestive system was observed and assessed whether the components would reach the crucial point of action – the colon. For the use of the proposed probiotic mixture as a dietary supplement, a recommended dose of 1 g has been determined. Lastly, possible adjustments such as lyophilization or sheathing by an extra protective polymerous layer, e.g. chitosan, were suggested to prolong the shelf life of the particles and volatile substances stability.
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