This diploma thesis deals with sweet substances from Stevia rebaudiana and with their determination in beverages. The ambition was to adjust the separation method for major steviol glycosides (stevioside and rebaudioside A) by HPLC so as to be fast and effective enough, too. In experimental part was successfully developed the separatin method, so it is possible to determine the steviol glycosides and conservation substances, specifically benzoic and sorbic acid (eventually their sodium or potassium salts). At first, for analysis was used monolithic column, on which stevioside and rebaudioside A weren´t separated. Other analyses were performed on column Ascentis express C18 (150 x 4.6; 2.7 µm). The most suitable separation method was performed by mobile phase made up with 29:71 v/v mixture of acetonitrile and acetate buffer (with pH 4.2). The flow rate was 0.28 ml/min and the column temperature was kept constant at 50 °C. The analysis of steviol glycosides and conservation substances with these condition was verified on the samples of real beverages.

Comparative studies of taste perception have found that primates may differ markedly in their sensitivity for substances perceived as sweet by humans. These findings raise questions about the reason that may underlie these differences in sweet-taste sensitivity between species. The aim of the present study was to assess the taste responsiveness of chimpanzees (Pan troglodytes verus) to seven substances tasting sweet to humans and to compare the results with those of other primate species. Using a two-bottle preference test (1 min) I found that the taste preference thresholds of the chimpanzees for five food-associated carbohydrates ranged between 20-30 mM for sucrose, 20-50 mM for fructose, 60-80 mM for glucose, 50-80 mM for maltose, and 30-80 mM for lactose. Taste preference thresholds for two steviol glycosides ranged from 0.04-0.05 mM for stevioside, and 0.03-0.05 mM for rebaudioside A. The chimpanzees displayed clear preferences for all sweet-tasting substances presented. In line with data obtained in other primates, the taste preference threshold of the chimpanzees for sucrose was lower compared to the other carbohydrates presented and the taste preference thresholds for stevioside and rebaudioside A were lower compared to sucrose. In general, the taste sensitivity of the chimpanzees fell into the range of data reported in other nonhuman primate species. Interestingly, the taste preference thresholds of the chimpanzees reported here are similar to the taste detection thresholds obtained in humans, despite the fact that the former are only a conservative approximation of an animal’s taste sensitivity. This suggests that chimpanzees may be as sweet-taste sensitive as humans.

Dental caries affects people worldwide. The oral microflora has been found to be less responsible than the carbohydrate diet, which is considered the main cause of dental caries. Carbohydrates from sucrose are processed by oral bacteria and acids are produced as a byproduct. High sugar intake is a risk factor for systemic health and for overall health of the oral cavity. Artificial sweeteners are considered a healthy alternative for people who prefer the sweet taste but renounce sucrose. In the United States there are several artificial sweeteners approved by the Food and Drug Administration. Among these, stevia is the only natural sweetener. Stevia is extracted from the Stevia rebaudiana plant. Steviol glycosides are extracted from the stevia leaf and refined to get the best taste quality. There are many types of steviol glycosides that are extracted from the stevia plant and the most important steviol glycosides will be discussed in the thesis. Scientific research (PubMed, EMBASE), appropriate websites (Colgate, Pure Circle Stevia Institute), the World Health Organization, HealthInfo Clinical Advisers and vectorstock as well as books in English and in Russian (The Stevia Deception: The Hidden Dangers of Low Calorie Sweeteners By Bruce Fife, and Puti Izbavlenie of Bolezne by Neumyvakin) were consulted. Electronic searches were limited to publications in the past five years. The scope of the thesis is to review the basic statistics on dental caries, the types of caries and the number of people affected, discuss the advantages and benefits of stevia relative to other artificial sweeteners, review the safety of stevia including side effects and potential as an addictive sugar substitute and lastly, to assess and recommend which form of stevia is most effective against dental caries.

The aim of the diploma thesis was evaluation of sensoric attributes of blueberry and current jams sweetened by selected Steviol glycosides and their comparing with other sweeteners. For evaluation of sensoric attributes were used rebaudioside A, stevioside, aspartame, erythritol and sugar beet. Stevia rebaudiana var. Bertoni is a persistent plant, which contains sweetening Steviol glycosides, which have a use in food processing. They are available in the area of EU indicated as E960 since 2011. The most used Steviol glycosides are stevioside and rebaudioside A, which are demarcated with a high sweetness, which could be 200-400 times higher than saccharose. The usage of sweeteners is legislatively allowed, which means it is also allowed to do sensoric analyse and detect perception of added substances in food. These analyses evaluate food with the help of sense - perception. Methodical part of the work contains procedure, used methods for gaining evaluation of data from a sensoric analyse after the degustation of jams, which has taken place with the presence of ten judges - an academic students in the working compartment of University of South Bohemia. Respondents entered their evaluation into protocols, which were prepared in advance. The results of diploma thesis are evaluated by the method of Kramer, which is used for this kind of analyse. The critical scale between 20 and 40 with the value of probability P = 95% was defined according to this method. Evaluation is described with the help of graphs and tables, added with a wordily describtion. Conclusion of analyse of diploma thesis is, that the usage of Steviol glycosides is not most suitable way of sweetening jams. Acceptable valuations of jams containing Steviol glycosides were proven in parameters of colour and aroma. The reasons for less successful evaluation of Steviol glycosides could be due to bad pH of food or badly chosen dosing, without combination of other kinds of sweeteners. The best sensory characteristics traditionally showed sugar beet and aspartame followed by rebaudioside A and stevioside, which were placed on the third and forth place from five used sweeteners. The part of the diploma thesis was an evaluation of questionnaire, where the result was, that asked people preferred fruit ingredient of jams (type of fruit, amount) not their way of sweetening while choosing a jam.

The steviolglycosides are the natural, sweet substances from Stevia rebaudiana Bertoni. It affect human health positively and its sweetness is 300x stronger than the sweetness of sucrose. That's why it's used to sweetening the commercial products. Because of its potential properties it's good to have an appropriate method to isolate it. High performance liquid chromatography ( HPLC) is based on the separation of analyte between two immiscible phases with high pressure pump and appropriately chosen stationary phase in the column. Than the analytes come out from the column in different retention times. This master´s thesis follows up selection of the best HPLC system for isolation of the main steviolglycosides and its analysis in commercial products. In the theoretical part of the thesis is described the origin, basic characteristics, botanical description, cultivation and affect to the human health of Stevia rebaudiana Bertoni and its use in food industry. There are also concisely characterized the sweet substances contained in the plant, so called steviolglycosides. Than there are given the theoretical basics of high performance liquid chromatography, HPLC instrumentation and its specific applications at sleviolglycosides with the basic chromatographic parameters. The object of the first experimental part was to research the optimum conditions for time and separation effective chromatographic analysis and select the best chromatographic system for isolation of steviolglycosides. In the second experimental part, I have compared and defined the main steviolglycosides (stevioside, rebaudioside A) in nine selected products, commercially available in Czech republic, with the best chromatographic method. In these products was the contain of the stevioside or rebaudioside A confirmed or refused.