The present study was aimed at understanding the high pressure (HP) processing-induced interactions of milk proteins in whey protein concentrate (WPC) solutions, in skim milk and in pure protein systems. The changes in milk proteins induced by heat treatments in the same systems under selected conditions were also evaluated. The main approach taken was to elucidate changes in the whey proteins in heat- and pressure-treated samples from common aliquots, under identical conditions, using various one-dimensional (1D) and two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE) techniques in the absence or presence of a disulphide bond reducing agent. In some instances, the samples were also analysed using small deformation rheology, size exclusion chromatography (SEC) and transmission electron microscopy (TEM). The results of the present study indicated that, in general terms, heat treatment and HP treatment had common effects, i.e. denaturation and subsequent aggregation of whey proteins. Both heat treatment and HP treatment generated disulphide-bonded and hydrophobically bonded aggregates of whey proteins. However, the sensitivities of each of the whey proteins to heat treatment [immunoglobulin (Ig) > lactoferrin (LF) > bovine serum albumin (BSA) > β-laetoglobulin B (β-LG B) > β-LG A > α-lactalbumin (α-LA)] and pressure treatment (β-LG B > β-LG A > IgG > LF > BSA > α-LA) were considerably different. Also, HP treatment generated a comparatively greater proportion of smaller aggregates than did heat treatment. The effects of protein concentration, intensity of pressure treatment, holding time and pressurising temperature on whey protein aggregation in WPC solutions were investigated. The rate of aggregation of whey proteins increased with an increase in the concentration of protein in the WPC solution and the pressurising temperature. The combination of low protein concentration, mild pressure treatment (200 MPa) and low pressurising temperature (20°C) led to minimal loss of native-like and SDS-monomeric β-LG, whereas the combination of high protein concentration, severe pressure treatment (600 MPa) and higher pressuring temperature (40°C and higher) led to significant loss of both native-like and SDS-monomeric β-LG. The sensitivity of pressure-resistant whey proteins, such as α-LA and BSA, to the aggregation was significantly increased at pressurising temperatures of 40°C and higher. Self-supporting gels were formed when 8 or 12% (w/v) WPC solutions were pressure treated at 600-800 MPa. 20°C. Detailed analysis of the behaviour of the proteins during the formation of these gels revealed a novel pathway, suggesting that intermolecular disulphide bond formation occurred at high pressure but that hydrophobic association became important after the HP treatment. In the later part of the study, heat- and HP-induced interactions of caseins and whey proteins were studied in a more complex system, i.e. skim milk. With the application of modified PAGE techniques, it was possible to show that the high molecular weight disulphide-bonded aggregates that were formed by HP treatment of skim milk contained disulphide-linked complexes consisting of αS2-casein (αS2-CN) as well as κ-CN, β-LG and other whey proteins. The results showed that the effects of heat treatment and HP on the interactions of the caseins and whey proteins in milk were significantly different. The accessibility of αS2-CN and the formation of complexes involving αS2-CN, κ-CN and whey proteins in the HP-treated milk, as demonstrated using the modified 2D PAGE technique, and as explained by possible proposed reactions of the caseins and whey proteins in pressure-treated milk, was an important finding of the present study. Finally, a study on the effects of HP treatment in model systems using pure proteins in solution, both singly or in binary and ternary combinations, generated very useful information and clarified the role of each protein in pressure-induced aggregation and interactions of milk proteins in complex systems such as WPC and milk. It was found that the reactions of β-LG were not significantly affected by other proteins such as α-LA or BSA, but that the presence of β-LG in the system catalysed the reactions of other proteins such as α-LA or BSA.

A shelf life trial using a fully balanced factorial experimental design was used to analyse the effects of acidity and light on broccoli juice made on a semi commercial scale over an eight week period in simulated retail refrigerated storage conditions. The research focused on making broccoli juice on a pilot scale, and what happens to the colour, composition and flavour during storage. A pilot scale production of pasteurised broccoli juice was conducted and the juice satisfied microbiological safety limits for the eight week shelf life trial in retail storage conditions. The stability of the green colour of fresh broccoli through processing and storage was assessed. Neutral broccoli juice remained green for four weeks before the colour became more yellow. The acidified juice became yellow on acidification and did not change significantly during storage. Dietary fibre and pectin levels did not change during storage. Chlorophyll and carotenoids levels decreased during storage and were directly influencing the colour changes in the juices. Ascorbic acid levels decreased significantly during processing resulting in low ascorbic acid levels (12 - 15 mg /100ml of juice) at the start of the shelf life trial and dropped further to 2-6 mg /100ml of juice after eight weeks. Acidification and storage in the dark had a protective effect on the degradation of ascorbic acid with only a 58% reduction in ascorbic acid levels compared to an 84% reduction in neutral light stored broccoli juice. The effect of processing and storage on the flavour of the beverage was assessed using a trained sensory panel providing descriptive analysis. The sensory profiles for neutral and acidified juices were extremely different with the unbalanced acidity suppressing the perception of the basic tastes, sweet, salty and bitter. The neutral juice sensory profile only changed slightly in aroma attributes during storage for seven weeks. The astringent aftertaste of the acidified juice increased while the broccoli smell decreased during storage. The results from this research indicate that the production of a broccoli juice with a yellow green colour and some retained nutritional components is achievable with a refrigerated (4 °C) shelf life of 30 days in light excluding glass packaging. The neutral juice is recommended as it was greener and had a broccoli flavour.A shelf life trial using a fully balanced factorial experimental design was used to analyse the effects of acidity and light on broccoli juice made on a semi commercial scale over an eight week period in simulated retail refrigerated storage conditions. The research focused on making broccoli juice on a pilot scale, and what happens to the colour, composition and flavour during storage. A pilot scale production of pasteurised broccoli juice was conducted and the juice satisfied microbiological safety limits for the eight week shelf life trial in retail storage conditions. The stability of the green colour of fresh broccoli through processing and storage was assessed. Neutral broccoli juice remained green for four weeks before the colour became more yellow. The acidified juice became yellow on acidification and did not change significantly during storage. Dietary fibre and pectin levels did not change during storage. Chlorophyll and carotenoids levels decreased during storage and were directly influencing the colour changes in the juices. Ascorbic acid levels decreased significantly during processing resulting in low ascorbic acid levels (12 - 15 mg /100ml of juice) at the start of the shelf life trial and dropped further to 2-6 mg /100ml of juice after eight weeks. Acidification and storage in the dark had a protective effect on the degradation of ascorbic acid with only a 58% reduction in ascorbic acid levels compared to an 84% reduction in neutral light stored broccoli juice. The effect of processing and storage on the flavour of the beverage was assessed using a trained sensory panel providing descriptive analysis. The sensory profiles for neutral and acidified juices were extremely different with the unbalanced acidity suppressing the perception of the basic tastes, sweet, salty and bitter. The neutral juice sensory profile only changed slightly in aroma attributes during storage for seven weeks. The astringent aftertaste of the acidified juice increased while the broccoli smell decreased during storage. The results from this research indicate that the production of a broccoli juice with a yellow green colour and some retained nutritional components is achievable with a refrigerated (4 °C) shelf life of 30 days in light excluding glass packaging. The neutral juice is recommended as it was greener and had a broccoli flavour.

Emulsions containing three different types of milk fat fractions (MF13, MF27 and MF42) and anhydrous milk fat (AMF) were prepared at oil to water (O/W) ratios of 1:9, 3:7, 5:5 and 7:3 using rhamnolipids as emulsifiers. The prepared emulsions were analyzed for their storage stability and properties (colour, particle size, zeta potential and rheology). The effects of various factors (freezing/thawing, heating, pH, salts and ionic strength) on the stability of emulsions were also investigated. All emulsions prepared with an O/W ratio of 7:3, regardless of the type of milk fat, rendered a highly condensed, semi solid and cream-like substance whereas other emulsions containing less oil were in a liquid form. Among the four different O/W ratios tested, the highest emulsion stability during the storage of 12 weeks was observed from the emulsions containing 1:9 O/W ratios, due to a combine effect of smaller emulsion particle size and lower collision frequency between droplets. Interestingly, the emulsions with 7:3 O/W ratios were found to be more stable than the ones with 5:5 O/W ratios. This might be due to the limited movements of closely-packed emulsion droplets induced by the high oil concentration of 7:3 O/W ratios. The emulsion stability was significantly affected by low pH, especially at lower than pH 4, due to the loss of electrostatic repulsions between droplets leading to droplet coalescence and also possibly due to hydrolysis of rhamnolipid molecules. The presence of salts (NaCl, KCl and CaCl2) also rendered the emulsion unstable. The degree of instability was gradually increased with increasing salt concentrations. CaCl2 had the most significant effect even at a very low concentration. The viscosity of emulsions increased with increasing oil concentration but was not affected by the types of milk fats. Emulsions with 3:7, 5:5 and 7:3 O/W ratios exhibited non-Newtonian and shear thinning flow behaviour. At 7:3 O/W ratios, MF13 exhibited gel-like properties whereas both MF42 and AMF emulsions became more solid-like at higher frequency.

Milk proteins are known to bind volatile flavour compounds to varying extents, depending on the nature of the protein and flavour compound. Processing conditions, such as temperature and pH, are also known to have an influence on the interactions between milk proteins and flavour compounds. These interactions cause a great challenge for flavour scientists because they influence the perceived aroma profile of food products significantly, in particular in low-fat food products. The objectives of this research were to develop a headspace solid-phase microextraction (SPME) method followed by gas chromatography with flame ionisation detection (GC-FID) for the investigation of protein-flavour interactions, and to determine binding parameters of the hydrophobic flavour compound, 2-nonanone, to individual milk proteins - namely, β-lactoglobulin (β-lg), α-lactalbumin (α-la), bovine serum albumin (BSA), αs1-casein, and β-casein -, whey protein isolate (WPI), and sodium caseinate. Secondly, it was the aim to compare the binding of the structurally similar flavour compounds - 2-nonanone, 1-nonanal, and trans-2-nonenal – to WPI in aqueous solution, and to investigate the effect of heat and high pressure treatment, and pH on the extent of protein-flavour binding. The final objective was to investigate the in vivo release of the reversibly bound flavour compound, 2-nonanone, from WPI and sodium caseinate using proton transfer-reaction mass spectrometry (PTR-MS), and to understand the effect of viscosity on flavour release in vivo. The binding of the model flavour compound 2-nonanone to individual milk proteins, WPI, and sodium caseinate in aqueous solutions was investigated, using headspace SPME followed by GC-FID. The 2-nonanone binding capacities decreased in the order: BSA > β-lg > α-la > αs1-casein > β-casein, and the binding to WPI was stronger than the binding to sodium caseinate. All proteins appeared to have one binding site for 2-nonanone, except for BSA which possessed two classes of binding sites. The influence of heat treatment, high pressure processing and pH of the protein solutions on the binding of 2-nonanone, 1-nonanal, and trans-2-nonenal to WPI was determined. The binding of these compounds to WPI decreased in the order: trans-2-nonenal > 1-nonanal > 2-nonanone. The binding of 2-nonanone appears to involve hydrophobic interactions only, whereas the aldehydes, in particular trans-2-nonenal, also react through covalent binding. Upon both heat and high pressure denaturation, the binding of 2-nonanone to WPI decreased, the binding of 1-nonanal remained unchanged, while the binding of trans-2-nonenal increased. The binding affinity of the flavour compounds and WPI increased with increasing pH, which is likely to result from pH dependent conformational changes of whey proteins. The in vivo flavour (2-nonanone) release from solutions of WPI and sodium caseinate was investigated using proton-transfer-reaction mass spectrometry. During consumption, 2-nonanone was partly released from WPI, whereas there was no significant release from sodium caseinate. Even after swallowing of the samples, a substantial amount of flavour was detected in the breath, suggesting that the milk proteins interact with the mucosa in the mouth and throat, resulting in a further release of flavour from mucosa-bound proteins. An increase in viscosity of the protein solutions by the addition of carboxymethylcellulose enhanced the release of 2-nonanone from WPI, and resulted in 2-nonanone release from sodium caseinate. This may be due to a thicker coating of the mucosa with the sample solution after swallowing due to the higher viscosity, resulting in additional release of protein-bound flavour. These findings contribute to the knowledge of the interactions that occur between flavour compounds and proteins, which is required to improve food flavouring and to make protein based foods, e.g., low-fat dairy products, sensorily more acceptable to the consumer. The results also emphasize a careful choice of food processing conditions, such as temperature, high pressure or pH to obtain a desirable flavour profile.

Made available in DSpace on 2014-10-09T12:37:50Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:56:13Z (GMT). No. of bitstreams: 1 05375.pdf: 5378656 bytes, checksum: 1bcd4f740c38a0583b83231181ad86b3 (MD5) / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

The aim of this thesis is to analyze the conditions under which a company can enter a foreign market. I have chosen the company Ipema, spol. s r. o. for my thesis. The company operates in food processing industry, specifically in the manufacture of bakery, since 1992. The company wants to expand it´s products to the Austrian market. Motivation and the reason for this process is the interest of Austrian customers in bread. To succeed in a foreign market is not easy. It is very important to know all the conditions and obligations which must be realize.

Submitted by ÉRICA AMANDA DE BARROS null (erica27ab@gmail.com) on 2016-03-17T21:03:52Z No. of bitstreams: 1 tese completa- ficha catalografica e ass da banca.pdf: 1132305 bytes, checksum: 5d57fcba149b07374e3b66cf5d039104 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-03-18T19:31:19Z (GMT) No. of bitstreams: 1 barros_ea_dr_bot.pdf: 1132305 bytes, checksum: 5d57fcba149b07374e3b66cf5d039104 (MD5) / Made available in DSpace on 2016-03-18T19:31:19Z (GMT). No. of bitstreams: 1 barros_ea_dr_bot.pdf: 1132305 bytes, checksum: 5d57fcba149b07374e3b66cf5d039104 (MD5) Previous issue date: 2016-01-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A irradiação é uma técnica de conservação ainda pouco aceita para alimentos e bebidas no Brasil, devido a grande parte da população desconhecer seus mecanismos de ação. Dessa forma, o objetivo desse estudo foi avaliar os aspectos físico-químico, bioquímico, microbiológico e sensorial de bebida mista de extrato hidrossolúvel de soja (EHS) e suco de uva, submetida à radiação gama. Neste estudo, foram realizados quatro experimentos, visando selecionar o melhor cultivar de soja (BRS-213, BRS-258 e EMBRAPA-48) em relação às características bioativas após a aplicação de diferentes doses de radiação gama (experimento 1). No segundo experimento, selecionou-se o método mais apropriado de produção do EHS a partir de duas formas de extração; determinou-se também a dose de radiação adequada para se aplicar ao EHS (etapa 1) e ao suco de uva (etapa 2), no experimento 3. No experimento 4 foi determinada a temperatura ideal de armazenamento e a adição ou não do conservante ácido benzoico em bebida mista de EHS e suco de uva. Para o experimento 1, realizou-se análises bioquímicas, sendo que nos demais, foram realizadas análises físico-químicas, bioquímicas, microbiológicas e sensoriais. No experimento 4, realizou-se também o teste de vida de prateleira. A partir dos resultados submetidos à estatística descritiva e inferencial (testes paramétricos - t e ANOVA; e testes não paramétricos -Kruskall-Wallis e Friedmann) constatou-se que a elevação da dose de radiação gama aumentou o teor dos compostos fenólicos para os cultivares de soja BRS-213 e BRS-258, bem como a atividade da enzima superóxido dismutase no cultivar BRS-213 (experimento 1). O EHS extraído a frio apresentou maior teor de proteína, lipídios e compostos fenólicos que o EHS extraído a quente. Por outro lado, a segunda extração apresentou melhores resultados na análise sensorial (experimento 2). No experimento 3 o EHS (etapa 1) submetido a 8 kGy apresentou menor teor de proteína e de compostos fenólicos. O EHS e o suco de uva (etapa 2) submetidos a 8 kGy demonstraram variação na cor e redução das notas do atributo sabor quando comparados aos demais tratamentos. A partir da dose de 2 kGy eliminou-se os bolores e leveduras do suco de uva. A bebida mista refrigerada e com adição de ácido benzoico apresentou maior vida de prateleira (9 meses). Concluiu-se que a dose de 4 kGy associada a refrigeração e ao conservante foi indicada para bebida mista de EHS com suco de uva, preservando suas características fitossanitárias, físico-químicas, bioquímicas e sensoriais. / The gamma irradiation is a conservation technique still not accepted for food and beverages in Brazil, due to much of the population ignore their mechanisms of action. Thus, the aim of this study was to evaluate the physic-chemical, biochemical, microbiological and sensory aspect of mixed beverage of soybean hydrosoluble extract and grape juice, submitted to gamma radiation. In this study, four experiments were conducted in order to select the best soybean cultivar regarding the bioactive characteristics (BRS-213, BRS 258 and EMBRAPA-48) after application of different doses of gamma radiation (experiment 1). In the second experiment, we selected the most appropriate method of producing soybean soybean hydrosoluble extract (EHS) from two forms of extraction. Also determined the adequate dosage of radiation to be applied to the EHS (step 1) and grape juice (step 2), in experiment 3. And the experiment 4 was determined the ideal storage temperature and with or without addition of the preservative benzoic acid in the mixed beverage of EHS and grape juice. For the experiment 1 was performed biochemical analyzes, and in the other were conducted in physical-chemical, biochemical, microbiological and sensory analiyses. In experiment 4, it was also carried shelf life test. When results submitted to descriptive and inferential statistics (tests parametric- t and ANOVA, and nonparametric tests -Kruskall-Wallis and Friedman) it was found that the elevation of gamma radiation dose increased the content of phenolic compounds for the soybean cultivars BRS-213 and BRS-258 and SOD activity in cultivar BRS-213 (experiment 1). The EHS cold extraction has a higher protein, lipid and phenolic compounds content that the EHS hot extraction. On the other hand, the second extraction showed better results in sensory analysis (experiment 2). In experiment 3 EHS (step 1) submitted to 8 kGy showed lower protein and phenolic compounds content. The EHS and grape juice (step 2) submitted to 8 kGy showed variation in color and reduction of notes of flavor attribute when compared to other treatments. From the dose of 2 kGy eliminates molds and yeast of grape juice. The refrigerated mixed beverage and with the addition of benzoic acid had longer shelf life (9 months). It was concluded that the dose of 4 kGy associated with refrigeration and preservative was nominated for mixed beverage of EHS with grape juice, preserving their phytosanitary characteristics, physical-chemical, biochemical and sensory.

Made available in DSpace on 2014-10-09T12:52:59Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:58:13Z (GMT). No. of bitstreams: 0 / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

Made available in DSpace on 2014-10-09T12:42:37Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:59:44Z (GMT). No. of bitstreams: 0 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Tese (Doutorado em Tecnologia Nuclear) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP / FAPESP:10/52170-9

Made available in DSpace on 2014-10-09T12:26:42Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:04:31Z (GMT). No. of bitstreams: 0 / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP