A new method for manufacture of natural cheese was developed using 5X ultrafiltered whole milk retentate. The retentate was prefermented to pH 5.0-5.2 before curd formation to simplify the cheese making process. It was demonstrated that the process can be made commercially feasible. Ultrafiltration and diafiltration of whole milk were done so that the desired level of residual lactose was left in the retentate. Retentate was inoculated with lactic starter culture and incubated (prefermented) until all lactose was converted to lactic acid. The final pH stabilized at about 5.0-5.2; the desired pH of the cheese curd. Incubation at 25°C allowed the retentate to remain liquid during prefermentation and easily pumped through pipes. After prefermentation, retentate was passed through a mechanical curd former where rennet was injected and mixed. The retentate-rennet mixture coagulated as it traveled upward through a cylinder. The curd emerged from the curd former and was cut into cubes by a grid of knives. The curd cubes were treated for removal of moisture by cooking in liquid and evaporation under vacuum. After moisture removal, curd was drained. salted and pressed. Cheddaring and milling were unnecessary. Experiments were performed to determine proper methods for preparing retentate. Diafiltration level was significantly related to lactose concentration in retentate (p=.OOO 1) and final pH of fermented retentate (p=.OOO1). Acidified permeate and acidified, deionized water were evaluated as cooking liquids. Cheese made from curd cooked in permeate had acid defects. but curd cooked in water produced cheese with a pH similar to conventionally produced Cheddar cheese. Moisture content of all finished cheese was excessive for Cheddar cheese. Body and texture of cheese made from curd cooked in water was close to conventionally produced Cheddar cheese. Diafiltration may be used to control final pH of fermented retentate. Prefermentation of retentate prior to cheese making will simplify equipment and shorten manufacturing time. Commercial application of the process is discussed.

ABSTRACT Effects of Filtration Temperature and Heat Treatment on Composition and Rheological Properties of Whole Milk Ultrafiltration Retentates John William Montella For the first part of my thesis, the effects of filtration temperature and heat treatment on the compositional properties of whole milk Ultrafiltration retentate (UF) were studied. Ultrafiltration is primarily run at temperatures in the range of 50-55°C but more and more plants are starting to filter at refrigeration temperatures. In the ultrafiltration of milk, filtration temperature can affect the composition of the retentate by affecting the chemistry of milk components. The application of a pasteurization step can also affect the chemistry of milk components. There were two filtration temperatures used: 10°C and 50°C. The effect of stage in the filtration process in which the pasteurization step is applied (before UF vs. after UF) is also studied. The heat treatment used was a batch pasteurization treatment of 63°C for 30 minutes. The milk was concentrated to a Volume Concentration Ratio (VCR) of 3X through a 10,000 Molecular Weight Cut Off polysulfone membrane. Compositional analysis was performed on permeate and retentate. According to my results, there were significant treatment effects on the retention of true protein (both casein and whey protein nitrogen), total protein, non-casein nitrogen, minerals (including Ca) and pH of the retentate. The chemistry of the milk components were considered as possible reasons for these differences. The week of processing did not affect the results. For the second part, the effect of composition of the retentate on their viscosity and flow properties was observed. Rheological properties are very important in process design and for consumer acceptability. Flow and viscosity data was collected using a dynamic stress rheometer. Three analytical temperatures were used during the rheological measurements: 10°C, 40°C, and 70°C. A shear rate of 500 s-1 was used for viscosity analysis. Flow properties were also observed using the same three temperatures. According to the results, all the retentate displayed shear thinning behavior and this behavior became more pronounced as the testing temperature increased. As the shear rate increased, there was a shear thickening effect that became more pronounced as temperature increased. There was a significant effect of treatment on the viscosities of the retentate. Compositional differences in the retentate are possible contributors to observed results. The week of processing had no effect on the results. For the final part, the effect of filtration temperature and heat treatment on rennet coagulation time of retentate was observed. A 22μl aliquot of chymosin was added to 100 ml of retentate heated to 30°C prior to analysis. Rennet coagulation time was monitored using a dynamic stress rheometer. The rennet coagulation time was recorded as the time at which the G’ value reached 1 Pa. There was a significant effect of filtration temperature and heat treatment on the rennet coagulation time of the retentate. Compositional differences are all possible contributors to these differences. From the observations from all three studies, the following conclusions can be made: (1) There were significant differences observed with respect to filtration temperature and heat treatments on chemical composition of the retentate; (2) The retentate displayed a shear thinning behavior and the chemical composition of the retentate could be a contributing factor as well as the sample testing temperature. There was also a significant treatment effect on the viscosity of the retentate; and (3) Significant differences in rennet coagulation times were observed, possibly due to compositional differences of retentate. Processing week did not have a significant effect on my results.

The aim of this project was to apply statistical process control (SPC) and measure the variation of fat content in milk in order to improve the standardization, so that the fat content does not change more than 0.03 percentage units from target. Recommendations of how to adjust the standardization should also be developed. The standardization takes place together with pasteurization in one of the three pasteurizers. Thereafter, the milk goes to a common product tank with all the pasteurizers. Samples from the three pasteurizers and the product tank were collected and analyzed on MilkoScan FT2 and the fat content was plotted into Shewhart and cumulative sum (CUSUM) charts. Sampling on the pasteurizers from startup showed that samples should be taken after about 20 minutes, because then the variation is in general smaller. The data from the product tank showed a smaller variation than the pasteurizers. Because the milk from all the pasteurizers is transported into one product tank, it is impossible to know which pasteurizer that is out of control and need to be adjusted. Therefore, the conclusion is that samples should be taken after the pasteurizer and plotted into Shewhart and CUSUM charts. Action limits were achieved from the Shewhart and CUSUM charts, respectively. These are the limits that should be used to determine when adjustments of the pasteurizers are needed, and not the brand limits that are considerably wider. If the measurements fall outside the second limit in the Shewhart chart (three times the standard deviation) or outside the limits (H) in the CUSUM chart, the standardization before the pasteurizer in question should be considered. It is not known if using SPC will improve the fat content to be within 0.03 percent units from target, because the recommendation has not been applied in the process yet, but it going to be that soon. / Syftet med detta arbete var att implementera statistisk processtyrning och mäta variationen i mjölkens fetthalt för att kunna förbättra standardiseringen. Detta skulle medföra att fetthalten inte förändras mer än 0,03 procentenheter från målet. Rekommendationer om hur standardiseringen bör justeras av fetthalten i mjölk ska också utvecklas. Standardiseringen sker tillsammans med pastöriseringen i en av de tre pastörerna. Därefter går mjölken till en gemensam produkttank för pastörerna. Prover från de tre pastörerna samt produkttank samlades in om analyserades på MilkoScan FT2 och fetthalten plottades in i Shewhart och Kumulativsumma (CUSUM) diagram. Provtagning från produktstart på pastörerna visade att prover skulle tas efter 20 minuter, eftersom variationerna då är mindre. Data från produkttank visade en lägre variation än hos de tre pastörerna. Eftersom mjölken från pastörerna transporteras till en produkttank är det omöjligt att veta vilken av de tre pastörerna som är utom kontroll och bör justeras. Därför är slutsatsen att proverna ska tas efter pastörerna och plottas in i Shewhart och CUSUM diagram. Åtgärdsgränser togs fram för både Shewhart och CUSUM diagram. Dessa är gränserna som ska användas för att bestämma när justeringar bör göras på pastörerna och inte varumärkesgränserna som är avsevärt bredare. Om ett mätvärde faller utanför den andra gränsen i Shewhart diagrammet eller utanför gränserna (H) i CUSUM diagrammet bör standardiseringen innan pastören i fråga justeras. Det är ännu oklart om standardiseringen kommer förbättras eftersom rekommendationerna inte har applicerats i processen än. Men kommer att göra det i den närmsta framtiden.

The heat-induced gelation properties of ultrafiltered (UF) whole milk concentrate were studied under different physical and chemical conditions. total solids concentration, homogenization pressures, heating temperatures, and heating times were found to have a positive correlation with gel strength. The addition of calcium chloride, sodium chloride, or trisodium citrate produced gels of higher strengths and textural properties than the gels obtained with non-salt-treated concentrate. Calcium chloride produced the strongest gels with a cheese-like texture and poor spreadability. Sodium chloride produced gels of intermediate strength with a firm, elastic texture and poor spreadability. Trisodium citrate produced the softest gels with a smooth, creamy texture and good spreadability. A shelf stable 40% total solids UF concentrate was manufactured using ultra-high temperature (UHT) processing by direct steam injection. The pourable concentrate had a shelf life of 75 to 90 days at 23°C and did not have the ability to produce heat-induced gels after a second heating. Addition of calcium chloride, sodium chloride, or trisodium citrate restored the heat-induced gelation of the retentate. However, the gels were weaker and presented different characteristics than did the gels from non-UHT-treated concentrate. Transmission electron microscopy (TEM) studies revealed a relationship between gel firmness and gel ultrastructure of the heat-induced gels. The gels consisted of a network of casein micelles connected with strands of a less dense protein material. The tighter the network the stronger the gel strength. High heating temperatures and calcium chloride addition caused fusion of the casein micelles int he network. Sensory evaluation of two prototype gelled desserts by a general consumer population showed a good potential for the use of the heat-induced gelation property of UF-concentrated whole milk in the development of new gelled dessert applications.

Three different concentrated whole milks (2.5x, 2.75x, and 3.0x) were produced by mixing equal parts of ultrafiltration retentate of whole milk and reverse osmosis retentate of the UF milk permeate. The concentrated whole milks were ultra-high temperature processed by direct steam injection (140.6°C) followed by flash cooling and two-stage homogenization pressures (2500/500 psi, 3500/700 psi, or 4500/900 psi). The milk concentrates were packaged aseptically and stored at room temperature. On the other hand, the milk concentrates produced by the RO single membrane system with the same concentration levels served as the control. Physicochemical properties of the milks were surveyed every 2 weeks during a 6-month storage period. The milk concentrates combined from the blending of multiple-membrane retentates showed the expeected concentrations of all major nutrients except nonprotein nitrogen. A 20% to 32% shortage of nonprotein nitrogen permeated through the RO membrane during the production of the concentrated whole milks. Over the 6 months' storage, nonprotein nitrogen content did not significantly change in the 2.5x, 2.75x, and 3.0x concentrated whole milks. No microbial growth or enzyme activity was measured or observed in the samples collected. Milk concentrated 2.5x with 4500/900psi homogenization pressure did not show cream plug formation during the first 5 months of storage. Milk concentrated 2.75x with 4500/900 psi homogenization pressure had the approximate cream plug level of the 2.5x concentrated milk at 4 months of storage. Milk concentrated 3.0x with 4500/900 psi homegenization pressure showed cream plugging at 2.5 months. As higher homogenization pressure was applied to the milk concentrates, less creaming occurred at all milk concentration levels. Homogenization at all pressures did not reduce or eliminate sedimentation during storage. The milk concentrates from the control RO membrane processing showed less sedimentation than did the concentrates from the multiple membrane system at the same homogenization pressure (2500/500 psi). the higher the concentration of total milk solids, the more sedimentation occurred. Viscosity was not affected by homogenization pressure in any of the concentrated whole milks.

Methods for on-farm extraction of low-concentration (minor) proteins from raw whole bovine milk directly after milking were explored. These minor proteins have high commercial value. Lactoferrin (LF) and lactoperoxidase (LP) were used as model proteins for extraction using cation exchange chromatography. Laboratory fractionations showed that milk could be processed by conventional column chromatography without excessive column backpressures if resin with large particles sizes were used and the temperature was high enough so fat in the milk was malleable; ideally the milk should be near the secretion temperature of 37oC. Processing parameters such as equilibrium and dynamic capacities were determined for SP Sepharose ™ (GE Healthcare Technologies) and Bio Rex 70 (BioRad Laboratories) resins. SP Sepharose Big Beads (SP BB) were found to be more suitable than BR 70, for raw whole milk processing due to the larger size (200 um). Design considerations showed that column chromatography was not the most practical method for on-farm processing of fresh, raw whole milk. Trials with a single-stage stirred tank showed that SP BB resin could extract up to 65% of LF (initial LF concentration of 0.5 mg/mL) with a 10-minute adsorption time. The composite non-linear (CNL) model of Rowe et al. (1999) was used to describe LF uptake by SP BB resin in raw whole milk with initial LF concentrations of 0 to 1.0 mg/mL and resin:milk volume ratios of 0.010, 0.012, 0.017 and 0.024 over 45-minute contact times. The CNL model could be used to predict LF yields if initial feed concentration, milk and resin volumes, and contact times were known. Laboratory extractions showed that processing did not significantly affect bulk milk composition (fat, protein, lactose and total solids), indicating that the milk could be used for conventional processing after the minor proteins had been extracted. Resin cleaning and regeneration studies, using a procedure similar to that recommended by the resin supplier, showed that the Sepharose resin had not degraded and there was no significant decrease in binding capacity after 50 extraction cycles. A Protein Fractionation Robot (PFR) prototype based on a single-stage stirred tank and the operating parameters obtained from the laboratory trials was designed, assembled and coupled to an Automated Milking System (AMS) to process fresh, raw whole milk from individual cows immediately after milking. The LF and LP extracted from the milk from 16 individual cows were 19.7 - 55.2% (35.6 10.2%) and 21.2 - 99.5% (87.1 12.0%) respectively. Generally, higher extraction levels were obtained at higher resin:milk ratios. The amount of LF extracted on-farm agreed within 14.1 9.8% of those predicted by the CNL model, with predicted values generally being higher. The experimental on-farm adsorption values were calculated using data of LF recovered after elution, so differences between actual and predicted values may be due to losses during post-adsorption processing. Economic feasibility studies, based on experimental data from the PFR and realistic wholesale prices for LF and LP ($400 and $150/kg respectively) showed that PFR-based processing is economically viable if the farmer is paid for the LF and LP produced as well as the bulk milk. This system would have a payback period of approximately five years and an internal rate of return of 14.5%. Further case studies determined the sensitivity of the economics to various operating parameters and value/cost assumptions, including producing recombinant human protein from transgenic bovine milk. These studies showed that the higher the value of the processed raw milk, the higher the absorptive capacity of the resin, and the higher the value of the extracted protein, the more favourable the economics. In the extreme case of producing a very high value therapeutic protein (e.g. $20 000), the payback period could be as low as 0.3 years, with an internal rate of return of 818%.

Manufacture of white soft cheese from ultrafiltered whole cows• milk involved acidification of pasteurized homogenized whole milk to pH 6.0 with phosphoric or citric acid. The preacidified milk was ultrafiltered at 54 °C until 60% of original milk weight was removed as permeate, diafiltered with deionized water equal to 38.5% of the original milk and concentrated by UF (4.8 fold) to pre-cheese (38% total solid). The pre-cheese was heated to 76.7°C/16 sec, 71 .l°C/l6 sec (HTST) and 7l.l°C/l5 min (controlled water bath), inoculated with 2% starter culture (Streptococcus cremoris), renneted (10 ml/100 lb retentate) and placed in one pound plastic containers in which coagulation took place (8-10 min). Salt (1 .5%) was added on the top of parchment paper placed under the lid, and the curd was incubated at 85°F. The most acceptable cheese was from ultrafiltered retentate heated for 16 sec at 76.7°C before cheese making. Organoleptic tests showed that samples highest in calcium content ranked highest in acceptability. Acidification with citric acid removed more calcium than phosphoric acid and resulted in softer cheese than the control cheese (non-acidified). Slight bitterness was observed when excessive starter and low salt (NaCl) concentration were used. Addition of salt to the retentate prior to heating caused thickening of the retentate before 70°C was reached. Extending the heating time increased the tendency toward mealiness in the cheese.

Sample preparation is an essential component of analytical methods in chemistry. It is not only necessary but also presents an opportunity to increase the effectiveness of the method significantly. There are various commercially available technologies for sample preparation, including numerous variations of LLE, SPE, and SPME. However, these technologies all present significant deficiencies, including the inability to extract directly from complex samples such as whole milk. Instrumental analysis has been improved greatly in the last two decades but still is not applicable to complex samples without sample preparation. This work presents the theory of FPSE, including the synthesis of sol-gel sorbents, coating of FPSE cellulose substrates and the mechanism of retention. Original research data presented herein introduce a comprehensive view on possible applications of FPSE in forensic chemistry and otherwise. Five distinct FPSE-based methods were rigorously developed for analysis of targeted pollutant residues. These methods were validated and compare to leading methods published in peer-reviewed literature quite favorably. Four of the methods were coupled to HPLC-UV and designed for trace or ultra-trace analysis of PAHs, BTEX, substituted phenols and nitroaromatic explosives, respectively. An additional FPSE-based method was developed and validated for direct analysis of BPA and five estrogenic EDCs in commercially purchased whole milk. This latter was coupled to both HPLC-UV and HPLC(QQQ)-MS/MS. The applicability of FPSE(PTHF) media was also tested for screening of aqueous samples and subsequent storage of analytes on the sorbent. My study included simultaneous extraction of a mixture of eight forensically significant compounds with various physicochemical properties and effective storage of each compound in frozen and ambient conditions for 32 weeks. These findings suggest that the storage ability of FPSE media can be extended as long as necessary, which is very significant in forensic laboratories where evidence often needs to be stored in a costly manner that may not be as effective in maintaining the chemical composition of the sample.

The polychlorinated biphenyls are synthetic organoclorated substances that can produce noxious effects to health. There are not any natural sources of PCBs known in the environment. They can be liquid or solid, colorless or clear yellow. Some of them are volatile and may remain as steams in the air. They were widely used in the industry as oils for transformers, electric capacitors, in transference fluids, inks, conservants for pesticides and lubricants, amongst others. Due to their inappropriate discard in the environment and also for the fact of presenting lingerence, bioaccumulation and high toxicity, they are distributed over the land surface, contaminating the alimentary chain. Victuals, mainly the most lipidic ones, such as the animal-originated products, have been recognized as the main source of PCBs ingestion. The concern about these compounds is in regard to their toxicological potential, mainly as cancer inductors. Milk and milk deriving products are an important source of macronutrients, with emphasis on proteins, vitamins, and mineral salts. Besides, they provide beneficial effects to health. Thus, the present dissertation has the purpose of investigating the existence of PCBs in integral powdered milk and to estimate the ingestion of these compounds from that victual by pre-scholars. Data of weight and consumption of powdered milk by children attending to infantile education schools of the municipal district of Imbé, RS, have been collected. The consumption of powdered milk among these children was of 32g per day, which is equivalent to 8,3g of daily fat. For the evaluation of the presence of PCBs 28, 52, 153, 138, and 180 in the powdered milk consumed by the pre-scholars we have proceeded the rebuilding of milk, followed by centrifugation in order to obtain the lipidic fraction. Subsequently, the lipidic content was extracted with sodium sulfate and ether of petroleum. The extraction and purification of the PCBs from fat were developed by using florisil and hexane. The identification and quantification of these compounds have been accomplished with the aid of the gas chromatograph equipped with micro electron capture detector (μECD). The confirmation was carried out on a gas chromatography coupled with mass spectrometer (GC/MS). The compounds concentrations were detected in the following order, from the most concentrated to the least concentrated: PCB 180 (0,157 ng g-1of fat) > PCB 138 (0,073 ng g-1) > PCB 28, 52 e 153 (<LQ). The PCB sum (PCB) was of 0,230 ng g-1of fat, oscillating from 0,000 to 2,857 ng g-1. The estimated daily ingestion of PCBs (PCBs) from powdered milk was of 0,0013 ng g-1 fat. This value, when compared with the reference limit admitted by the Brazilian legislation, is under the value established. On the basis of the results obtained it was possible to conclude that the milk consumed by the pre-scholars from PNAE of the municipal district of Imbé does not offer any toxicological risk. Nevertheless, it is suggested the importance of other works on this area with the purpose of monitoring the presence of residues of PCBs in milky products addressed to the infantile population once that they can generate adverse effects to health. / Os bifenilos policlorados são substâncias organocloradas sintéticas que podem causar efeitos lesivos à saúde. Não existem fontes naturais conhecidas de PCBs no ambiente. Eles podem ser líquidos ou sólidos, incolores ou amarelo claro. Alguns são voláteis e podem permanecer como vapores no ar. Foram amplamente utilizados na indústria como óleos para transformadores, capacitores elétricos, em fluídos de transferência, tintas, conservantes para pesticidas, lubrificantes e etc. Devido ao seu descarte inadequado no meio ambiente, e por apresentarem persistência, bioacumulação e grande toxicidade encontram-se distribuídos na superfície terrestre, contaminando a cadeia alimentar. Alimentos, especialmente os mais lipídicos, como os produtos de origem animal têm sido reconhecidos como a principal fonte de ingestão de PCBs. A preocupação quanto a estes compostos está voltada para o seu potencial toxicológico, principalmente como indutores de câncer. Leite e derivados lácteos, constituem uma importante fonte de macronutrientes, em especial as proteínas, vitaminas e sais minerais. Além disso, proporcionam efeitos benéficos à saúde. Diante do exposto, a presente dissertação tem como objetivo investigar a presença de PCBs em leite em pó integral e estimar a ingestão destes compostos a partir deste alimento por pré-escolares. Foram coletados os dados de peso e consumo de leite em pó por crianças matriculadas nas escolas de educação infantil no município de Imbé RS. O consumo de leite em pó pelas crianças foi de 32 g-1 dia, equivalente a 8,3 g de gordura/dia. Para avaliar a presença dos PCBs 28, 52, 153, 138 e 180 no leite em pó integral utilizado pelo Programa Nacional de Alimentação Escolar (PNAE) de Imbé e consumido pelos pré-escolares da rede de ensino municipal, procedeu-se a reconstituição do leite, seguido da centrifugação para obtenção da fração lipídica. Posteriormente, o conteúdo lipídico foi extraído com sulfato de sódio e éter de petróleo. A purificação e extração dos PCBs, a partir da gordura, foram feitas utilizando-se florisil e hexano. A identificação e a quantificação destes compostos foram feitas com auxílio de cromatógrafo a gás dotado de microdetector de captura de elétrons (μECD). A confirmação foi realizada por cromatógrafo a gás acoplado à espectrometria de massas (GC-MS). As concentrações dos compostos foram detectadas na seguinte ordem, do mais concentrado para o menos concentrados: PCB 180 (0,157 ng g-1 de gordura) > PCB 138 (0,073 ng g-1) > PCB 28, 53 e 153 (<LQ). O somatório de PCB (PCB) foi de 0,230 ng g-1 de gordura, variando de 0,000 a 2,857 ng g-1. A ingestão diária estimada de PCBs (PCBs), a partir de leite em pó, foi 0,0013 ng g-1 de gordura. Este valor, quando comparado com o limite de referência admitido pela legislação brasileira fica abaixo do valor estabelecido. Com base nos resultados obtidos, conclui-se que o leite consumido pelos pré-escolares do PNAE no município de Imbé não oferece risco toxicológico. No entanto, sugere-se a importância de outros trabalhos na área, com a finalidade de monitorar a presença de resíduos de PCBs em produtos lácteos destinado ao público infantil, uma vez que estes podem provocar efeitos adversos à saúde.