The purpose of this research was to relate lactose reactivity and age gelation of UHT processed concentrated milk. Skim milk was pasteurized, diafiltered batchwise to reduce lactose concentration to less than 0.05%, and UF concentrated to 3X (one-third volume reduction). Lactose and sucrose were then each added at 3% or 6% w/v to part of the concentrate. The five samples, control ( < 0.05% lactose), 3% w/v lactose, 6% w/v lactose, 3% w/v sucrose, and 6% w/v sucrose, were UHT processed at 140°C for 4 s using the indirect heating method. Samples were collected aseptically in presterilized plastic containers and stored at 4°C, 20°C, and 35°C for periodic analysis. All samples stored at 4°C and 20°C gelled after 21 weeks of storage. The viscosity changed slightly during the first 19 weeks of storage but increased suddenly (> 100 cPs) just before gelation. Samples stored at 35°C did not gel but showed sedimentation. Samples stored at 4°C or 20°C underwent little browning; but samples containing 3% and 6% lactose, stored at 35°C, browned considerably. The SOS-PAGE patterns of gelled samples showed new bands because of proteolysis whereas samples stored at 35°C showed bands due to proteolysis and protein crosslinking and a streaking pattern. Electron micrographs of gelled samples showed various casein particles connected together by hairy appendages protruding from the surface of casein particles, to form a continuous three-dimensional network. In non-gelled samples, the micelles were not joined into a continuous network and few hairy appendages protruded from their surfaces. Hairy appendages were not a result of Maillard reaction occurring during storage. Maillard reaction neither provided protection against nor promoted age gelation. Proteolysis was not the only cause for gelation. Protein modifications prevented gelation in samples stored at 35°C. Age gelation was probably a two-step process involving dissociation of proteins from the casein micelles that reformed onto the micelle surface as hairy appendages. Aggregation of the protein particles occurred through these appendages rather than through the original micelle surface.

Structure function changes of casein micelles were studied as a function of concentration using a non invasive concentration method, osmotic stressing. A combination of serum analysis, light scattering and rheological measurements were used to characterize the physico-chemical properties of casein micelles. In heated and unheated milk, rheological studies indicated that casein micelles behave as hard spheres of similar volume fractions, if the viscosity changes in the serum phase and the particle particle interactions are taken into account. The differences in the distribution of the heat induced complexes between colloidal and soluble phase affected the colloidal properties of casein micelles. Above 70 g L-1 protein, the protein particles were no longer free diffusing. Re-dilution of the suspensions showed no irreversible aggregation. The data suggested that in the range of concentration studied casein micelles behave as hard spheres. Age gelation was also investigated on heated and unheated concentrated milk. In unheated concentrated milk proteolysis played an important role in imparting an increase in viscosity by causing aggregation of the casein micelles. On the other hand, in heated milk, there was a significant effect of the whey protein aggregates, which increased their interaction with the casein micelles over time. This effect, together with proteolysis caused age gelation in heated concentrated milk. The method of concentration used in this research, osmotic stressing, was then compared to ultrafiltration. It was demostrated that these two methods are not equivalent, as shear and mixing during ultrafiltration cause rearrangements to the casein micells. The differences were clearly demonstrated by adding soluble caseins to the milk before or after concentration. This project brings a better understanding on the effects of concentration on the structure-function of casein micelles and the interactions occurring in milk proteins during concentration.

The purpose of this research was to investigate the effect of ultra-high temperature process parameters on age gelation of milk concentrate. Skim milk was concentrated to 2X (volume reduction) using reverse osmosis. The milk concentrate was preheated at 75 or 90°C for 20 or 50 s and UHT-processed at 138 or 145°C for 4 or 16 s. Sterilizing methods used were direct steam injection and indirect plate heat exchanger. The samples were aseptically collected in presterilized plastic containers and stored at 15 or 35°C. At 15°C storage temperature, the steam-injected samples gelled in 5 months when 4 s UHT time was used. When UHT time was increased to 16 s, the samples gelled in 6 months. Of the samples that were UHT processed by indirect plate heat exchanger for 4 s and stored at 15°C, all gelled after 7 months. When UHT time was increased to 16 s, all the 138°C samples gelled after 7 months as did the samples that were preheated for 50 s and UHT-processed at 138°C. The samples preheated at 75°C for 50 sand UHT-processed at 145°C gelled after 8 months, whereas at 90°C preheat temperature the samples gelled after 9 months. The samples stored at 35°C did not gel but showed different sedimentation levels. The sediment depth in the container was always greater for the steam-injected samples. The samples that received higher heat treatments by the two processing methods had a higher sedimentation depth. The pH decreased during storage and the extent of reduction was higher at 35°C storage temperature . Maillard browning occurred at both storage temperatures. Browning was greater in samples stored at 35°C and processed by indirect plate heat exchanger.

The effect of ultra-high temperature processing by direct steam injection and room temperature storage of pH-adjusted and unadjusted 3X (volume reduction) skim milk retentate and the effect of storage at various temperatures of 3X skim milk retentate without pH adjustment on their ζ-potential, viscosity, and pH were determined. Pasteurized skim milk was concentrated to 3X by ultrafiltration. In the pH study portions of skim milk retentate were adjusted to pH 6.38 ± .02 with HCl and 6.85 ± .01 and 7.32 ± .02 with NaOH between ultrafiltration and ultra-high temperature processing. In the storage temperature study, storage temperatures used for pH-unadjusted retentate samples were 11°C, 23°C (room temperature), and 37°C. Although pH 6.38 samples had the lowest viscosity in the pH study before ultra-high temperature processing, these samples precipitated during ultra-high temperature processing. For non-acidified samples, increasing pH of retentate resulted in higher viscosities and quicker age gelation times. Destabilization occurred more rapidly at 37°C than at 23°C or 11°C. The pH drop tended to be greater for samples stored at a higher temperature or adjusted to a higher pH. During 28 weeks of 37°C storage the pH decreased from 6.54 to 6.06. During 32 weeks of 23°C storage pH of samples initially adjusted to pH 7.32 dropped to 7.06. ζ-Potentials of casein micelles were calculated from electrophoretic mobility obtained by measuring Doppler frequency shifts of scattered laser light in samples that had been diluted 300 fold with their own ultra.filtrate. Absolute values of ζ-potential of samples stored at 37°C decreased from -23.4 millivolts immediately after ultra-high temperature processing to -18.5 millivolts after 28 weeks of storage. For samples stored at 11°C and 23°C in the storage temperature study and control samples in the pH study, absolute values of ζ-potential decreased approximately 1.5 to 2.0 millivolts during 28 or 32 weeks of storage.

Orientador: Mirna Lucia Gigante / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-15T18:42:59Z (GMT). No. of bitstreams: 1 Vianna_PriscilaCristinaBizam_D.pdf: 1512890 bytes, checksum: cfe5342097a5fc9d5f272c136b132b45 (MD5) Previous issue date: 2010 / Resumo: O objetivo deste trabalho foi avaliar o efeito da adição de dióxido de carbono (CO2) sobre a qualidade dos leites cru e UHT. Inicialmente, o leite cru adicionado ou não de CO2 foi armazenado em garrafas de vidro a 4 e 7ºC e amostras foram avaliadas diariamente quanto à contagem padrão em placas, psicrotróficos e Pseudomonas spp. e a cada dois dias quanto à concentração de CO2, proteólise e lipólise, até que a contagem padrão em placas atingisse 7,5x105 ufc/mL. O delineamento experimental utilizado foi o de sub-sub-parcelas divididas e os resultados foram avaliados por análise de variância multivariada, pelo teste de médias de Tukey (p<0,05) e através do modelo matemático de Gompertz. A contagem padrão aumentou ao longo do tempo para todos os tratamentos e o tempo de conservação foi de 14 dias para o leite armazenado a 4ºC adicionado de CO2 e de 5 dias para o armazenado a 7ºC não adicionado de CO2. Independente da temperatura de armazenamento, a adição de CO2 estendeu o tempo de fase lag e de geração dos micro-organismos psicrotróficos e reduziu sua taxa de crescimento. O leite adicionado de CO2 apresentou menor proteólise e lipólise quando comparado ao não adicionado devido ao menor desenvolvimento de psicrotróficos. Com base nesses resultados, definiu-se o armazenamento do leite cru adicionado ou não de CO2 a 4ºC por 6 dias antes do processamento UHT. Nesta etapa, o leite cru adicionado ou não de CO2 foi armazenado em tanques de expansão antes do processamento UHT (140ºC/5 s). O leite cru foi avaliado no dia da recepção e após 6 dias de armazenamento quanto à composição físico-química, proteólise, lipólise e contagens microbianas. Após o processamento, as amostras foram avaliadas durante 120 dias quanto à composição físico-química, lipólise e proteólise. O delineamento experimental utilizado foi o de parcelas subdivididas em blocos com três repetições. Os resultados foram avaliados por análise de variância e teste de médias de Tukey (p<0,05). Após 6 dias de armazenamento a 4 ºC o leite cru adicionado de CO2 manteve sua qualidade físico-química e microbiológica enquanto que o leite não adicionado sofreu perda significativa de qualidade. A taxa de aumento de proteólise foi 1,4 vezes maior no leite UHT produzido a partir de leite cru não adicionado de CO2 quando comparado à amostra produzida com adição de CO2. Em ambas as amostras, a proteólise foi decorrente tanto da ação de plasmina como de proteases microbianas. Entretanto, o leite UHT produzido a partir de leite cru não adicionado de CO2 apresentou maior ação de proteases microbianas, caracterizada pelo aumento de peptídeos menos hidrofóbicos. A lipólise aumentou para as duas amostras durante os 120 dias e foi maior no leite UHT produzido a partir de leite cru não adicionado de CO2. Os resultados mostraram que a adição de CO2 ao leite cru preservou sua qualidade físico-química e microbiológica durante o armazenamento refrigerado e afetou positivamente a manutenção da qualidade do leite UHT durante seu armazenamento / Abstract: The objective of this work was to evaluate the effect of the CO2 addition on the raw and UHT milk quality. First, raw milk with or without CO2 addition was stored in glass bottles at 4ºC and 7ºC and daily analyzed to standard plate count, psychrotrophic bacteria count and Pseudomonas spp. and every other day to CO2 concentration, proteolysis and lipolysis until standard plate count reached 7,5x105 ufc/mL. Split-split-plot design was used and the results were evaluated by multivariate variance analysis, Tukey¿s test (p<0,05) and by Gompertz model. The standard plate count increased throughout the time for all treatments and the preservation time was 14 days for CO2 added raw milk stored at 4ºC and 5 days for raw milk without CO2 addition stored at 7ºC. Independent of the storage temperature, CO2 extended the lag phase, increased the generation time and decreased the growth rate of psychrotrophic bacteria. Milk with CO2 addition presented lower proteolysis and lipolysis, related to the slower psychrotrophic bacteria development. Based on these results, it was defined a storage of raw milk with and without CO2 addition in bulk tanks at 4ºC during 6 days before UHT treatment (140ºC/5 s). Raw milk was evaluated to physical-chemical composition, proteolysis, lipolysis and microbial counts in the day of reception and after 6 days of storage. After processing, samples were evaluated to physical-chemical composition, lipolysis and proteolysis. Split-plot design was used with three replications. The results were evaluated by analysis of variance (ANOVA) and Tukey¿s test (p<0,05). After 6 days of storage at 4ºC, CO2 added raw milk kept its physical-chemical and microbiological quality, while raw milk without CO2 addition had significant losses. Proteolysis increased ratio was 1,4 higher in UHT milk produced with raw milk without CO2 addition when compared to UHT milk produced with CO2. In both samples proteolysis was a consequence of plasmin and microbial proteases action, but the sample produced with raw milk without CO2 addition presented higher microbial proteases action, characterized for the increase of less hydrophobic peptides. Lipolysis increased for both samples during the 120 days of storage and it was higher in UHT milk produced from raw milk without CO2 addition. The results had shown that CO2 addition to raw milk preserved its physical-chemical and microbiological quality during refrigerated storage and positively affected the quality maintenance of UHT milk during its storage / Doutorado / Doutor em Tecnologia de Alimentos

Orientador: Mirna Lúcia Gigante / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-19T11:42:02Z (GMT). No. of bitstreams: 1 Dias_MariaElisabeteFernandesDias_M.pdf: 5400605 bytes, checksum: 00a236017885ffd7ef50a95f530931bb (MD5) Previous issue date: 2011 / Resumo: O objetivo deste trabalho foi avaliar o efeito da adição de CO2 ao leite cru sobre as características do leite UHT obtido por injeção direta de vapor e armazenamento a 25ºC, 35ºC e 45ºC por 180 dias. O leite cru (250 litros) foi dividido em duas porções que foram armazenadas em tanques de expansão a 4?1ºC por seis dias. Uma porção foi adicionada de CO2 grau alimentício até que o pH do leite atingisse 6,20, enquanto a outra serviu de controle. O leite cru foi avaliado quanto ao pH, acidez, prova do álcool, composição físico-química, proteólise, lipólise, cor e concentração de CO2 após a injeção. Para caracterização microbiológica, o leite cru foi avaliado quanto à contagem padrão em placas e de micro-organismos psicrotróficos no dia da recepção e após seis dias de armazenamento refrigerado. As amostras foram submetidas ao tratamento UHT por injeção direta de vapor (143ºC/4s), envasadas em embalagens tetra brik asseptic de 125 ml e armazenadas em BOD a 25, 35 e 45 ºC por 180 dias. No dia seguinte, as amostras foram avaliadas quanto as mesmas características do leite cru, além da prova do álcool, viscosidade, sedimentação, eletroforese, peptídeos por HPLC e esterilidade comercial. Após 1, 30, 60, 90, 120, 150 e 180 dias de armazenamento, as amostras foram avaliadas quanto ao pH, acidez, nitrogênio e frações nitrogenadas, cor, ácidos graxos livres, sedimentação, viscosidade, eletroforese e peptídeos por HPLC. O delineamento experimental foi o de sub-sub-parcelas divididas e o experimento foi repetido três vezes. O efeito do tratamento, da temperatura e do tempo de armazenamento, e a interação destes fatores sobre as características do leite UHT foi avaliado por análise de variância (ANOVA) e teste de Tukey ao nível de 5% de significância. Após seis dias de armazenamento refrigerado, não houve diferença significativa nas características físico-químicas do leite adicionado ou não de CO2, exceto na quantidade de ácido graxos livres que foi maior no leite controle do que no adicionado de CO2. A adição de CO2 inibiu o desenvolvimento de micro-organismos durante o armazenamento refrigerado, uma vez que a contagem total e de psicrotróficos do leite controle foi maior que as contagens do leite adicionado de CO2 . O pH das amostras de leite UHT armazenadas a diferentes temperaturas foi afetado pela temperatura e pelo tempo de armazenamento, apresentando maior decréscimo do pH nas amostras armazenadas a 45 ºC. A cor das amostras a 25°C não apresentou escurecimento durante os 180 dias, enquanto as armazenadas a 35°C e 45°C apresentaram desenvolvimento de cor visível a olho nu e aumento no valor b* ao longo do tempo. A lipólise do leite UHT foi maior nas amostras armazenadas a 45ºC. A proteólise foi maior no leite armazenado a 45°C, cujo aumento não refletiu na viscosidade e sedimentação do leite, que não foram significativamente afetados até o 120° dia de armazenamento, prazo de validade usualmente garantido pelas indústrias de processamento de leite UHT no Brasil / Abstract: The objective of this study was to evaluate the effect of CO2 addition to raw milk on the characteristics of UHT milk obtained by direct steam injection and stored at 25 ° C, 35 ° C and 45 ° C for 180 days. Raw milk (250 liters) was divided in two portions that were stored in bulk tanks at 4 ± 1 ° C for six days. To one portion was added food grade CO2 until the pH of milk was 6.20, while the other portion was the control sample. Raw milk was evaluated for pH, acidity, physicochemical composition, proteolysis, lipolysis, color and concentration of CO2 after injection. Raw milk was evaluated for standard plate count and psychrotrophic micro-organisms on the reception and after six days of cold storage, for microbiological characteristics. The samples were submitted to UHT treatment by direct steam injection (143°C/4s), packed in 125 ml Tetra Brik Asseptic packing and stored in BOD at 25, 35 and 45 ° C for 180 days. One day after processing, the samples were evaluated for the same characteristics of raw milk, plus alcohol stability, viscosity, sedimentation, electrophoresis, peptides by HPLC and commercial sterility. After 1, 30, 60, 90, 120, 150 and 180 days of storage, samples were evaluated for pH, acidity, nitrogen and nitrogen fractions, color, free fatty acids, sedimentation, viscosity, electrophoresis and peptides by HPLC. The experimental design was split-split-plot and the complete experiment was repeated three times. The effect of treatment, temperature and storage period as well as the interaction of these factors on the characteristics of UHT milk was assessed by analysis of variance (ANOVA) and Tukey¿s test at 5% significance level. After six days of cold storage, there was no significant difference in the physicochemical characteristics of raw milk with or without CO2 addition, except for free fatty acid values, which were higher for the milk without CO2 addition. The addition of CO2 inhibited the development of microorganisms during storage, since the standard plate count and psychrotrophic count of milk control was higher than values for the CO2 added milk. The pH of UHT milk samples was affected by temperature and storage time, showing greater decrease in pH for samples stored at 45°C. The color of the samples at 25°C showed no browning during the 180 days, while those stored at 35 ° C and 45 ° C showed brown color visible to the naked eye and an increased b * value over time. Lipolysis of UHT milk samples was higher for samples stored at 45°C. The proteolysis was higher in milk stored at 45°C, whose increase was not reflected in viscosity and sedimentation of the milk, since these parameters were not significantly affected during 120 days-storage, period of shelf life usually guaranteed by the processing industries of UHT milk in Brazil / Mestrado / Tecnologia de Alimentos / Mestre em Tecnologia de Alimentos