Physicochemical properties of micellar casein retentates generated at different microfiltration temperatures.

Processing temperature has a significant influence on the composition and functionality of the resulting streams following microfiltration (MF) of skim milk. In this study, MF and diafiltration (DF) were performed at 4 or 50°C to produce ß-casein (ß-CN)-depleted and nondepleted (i.e., native casein profile) micellar casein isolate retentates, respectively. Microfiltration combined with extensive DF resulted in a 40% depletion of ß-CN at 4°C, whereas no ß-CN depletion occurred at 50°C. Microfiltration at 4°C led to higher transmission of calcium into permeates, with retentate generated at 4°C containing less total calcium compared with retentate generated at 50°C, based on the volume of retentate remaining. Higher heat stability at 120°C was measured for retentates generated at 4°C compared with those at 50°C, across all pH values measured. Retentates generated at 4°C also had significantly lower ionic calcium values at each pH compared with those generated at 50°C. Higher apparent viscosities at 4°C were measured for retentates generated at 4°C compared with retentates generated at 50°C, likely due to increased voluminosity of ß-CN-depleted casein micelles. The results of this study provide new information on how changing the composition of MF retentate, by appropriate control of processing temperature and DF, can alter physicochemical properties of casein micelles, with potential implications for ingredient functionality.

Ice crystallization and structural changes in cheese during freezing and frozen storage: implications for functional properties.

Temperature-mediated preservation techniques offer a simple, scalable, effective, and fairly efficient method of long-term storage of food products. In order to ensure the uninterrupted availability of cheese across the globe, a critical understanding of its techno-functional properties as affected by freezing and frozen storage is essential. Detailed studies of temperature-mediated molecular dynamics are available for relatively simpler and homogeneous systems like pure water, proteins, and carbohydrates. However, for heterogeneous systems like cheese, inter-component interactions at sub-zero temperatures have not been extensively covered. Ice crystallization during freezing causes dehydration of caseins and the formation of concentration gradients within the cheese matrix, causing undesirable changes in texture-functional attributes, but findings vary due to experimental conditions. A suitable combination of sample size, freezing rate, aging, and tempering can extend the shelf life of high- and low-moisture Mozzarella cheese. However, limited studies on other cheeses suggest that effects and suitability differ by cheese type, in most cases adversely affecting texture and functional attributes. This review presents an overview of the understanding of the effects of refrigeration, freezing techniques, and frozen storage on structural components of cheese, most prominently Mozzarella cheese, and the corresponding impact on microstructure and functionality. Also included are the mechanism of ice formation and relevant mathematical models for estimation of the thermophysical properties of cheese to assist in designing optimized schemes for their frozen storage. The review also highlights the lack of unanimity in critical understanding concerning the effect of freezing on the long-term storage of Mozzarella cheese with respect to its functionality.

Molecular mechanisms contributing to the development of beef sensory texture and flavour traits and related biomarkers: Insights from early post-mortem muscle using label-free proteomics.

Beef sensory quality comprises a suite of traits, each of which manifests its ultimate phenotype through interaction of muscle physiology with environment, both in vivo and post-mortem. Understanding variability in meat quality remains a persistent challenge, but omics studies to uncover biological connections between natural variability in proteome and phenotype could provide validation for exploratory studies and offer new insights. Multivariate analysis of proteome and meat quality data from Longissimus thoracis et lumborum muscle samples taken early post-mortem from 34 Limousin-sired bulls was conducted. Using for the first-time label-free shotgun proteomics combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), 85 proteins were found to be related with tenderness, chewiness, stringiness and flavour sensory traits. The putative biomarkers were classified in five interconnected biological pathways; i) muscle contraction, ii) energy metabolism, iii) heat shock proteins, iv) oxidative stress, v) regulation of cellular processes and binding. Among the proteins, PHKA1 and STBD1 correlated with all four traits, as did the GO biological process 'generation of precursor metabolites and energy'. Optimal regression models explained a high level (58-71%) of phenotypic variability with proteomic data for each quality trait. The results of this study propose several regression equations and biomarkers to explain the variability of multiple beef eating quality traits. Thanks to annotation and network analyses, they further suggest protein interactions and mechanisms underpinning the physiological processes regulating these key quality traits. SIGNIFICANCE: The proteomic profiles of animals with divergent quality profiles have been compared in numerous studies; however, a wide range of phenotypic variation is required to better understand the mechanisms underpinning the complex biological pathways correlated with beef quality and protein interactions. We used multivariate regression analyses and bioinformatics to analyse shotgun proteomics data to decipher the molecular signatures involved in beef texture and flavour variations with a focus on multiple quality traits. We developed multiple regression equations to explain beef texture and flavour. Additionally, potential candidate biomarkers correlated with multiple beef quality traits are suggested, which could have utility as indicators of beef overall sensory quality. This study explained the biological process responsible for determining key quality traits such as tenderness, chewiness, stringiness, and flavour in beef, which will provide support for future beef proteomics studies.

A longitudinal study of fatty acid profiles, macronutrient levels, and plasmin activity in human milk.

Introduction: Human milk provides nutrients essential for infant growth and health, levels of which are dynamic during lactation. Methods: In this study, changes in macronutrients, fatty acids, and plasmin activities over the first six months of lactation in term milk were studied. Results: There was a significant influence of lactation stage on levels of protein and plasmin activities, but not on levels of fat and carbohydrate in term milk. Concerning fatty acids in term milk, levels of caproic acid and α-linolenic acid increased significantly (p < 0.05), whereas those of arachidonic acid and docosahexaenoic acid decreased, in the six months after birth. Significant impacts of maternal pre-pregnancy BMI and infant gender on fatty acid profiles were also found. Multivariate statistical analysis showed that protein level, plasmin activity, and several fatty acids (α-linolenic acid, lignoceric acid, and docasadienoic acid) contributed strongly to discrimination of milk from different lactational stages. Discussion: The study demonstrates that not all but some fatty acids were influenced by lactation, whereas protein and protease levels showed clear decreasing trends during lactation, which may help in understanding the nutritional requirements of infants.

Dynamic Changes in the Human Milk Metabolome Over 25 Weeks of Lactation.

Human milk (HM) provides essential nutrition for ensuring optimal infant growth and development postpartum. Metabolomics offers insight into the dynamic composition of HM. Studies have reported the impact of lactation stage, maternal genotype, and gestational age on HM metabolome. However, the majority of the studies have considered changes within the first month of lactation or sampled with large intervals. This leaves a gap in the knowledge of progressing variation in HM composition beyond the first month of lactation. The objective of this study was to investigate whether the HM metabolome from mothers with term deliveries varies beyond 1 month of lactation, during the period in which HM is considered fully mature. Human milk samples (n = 101) from 59 mothers were collected at weeks 1-2, 3-5, 7-9, and 20-25 postpartum and analyzed using 1H nuclear magnetic resonance spectroscopy. Several metabolites varied over lactation and exhibited dynamic changes between multiple time points. Higher levels of HM oligosaccharides, cis-aconitate, O-phosphocholine, O-acetylcarnitine, gluconate, and citric acid were observed in early lactation, whereas later in lactation, levels of lactose, 3-fucosyllactose, glutamine, glutamate, and short- and medium-chain fatty acids were increased. Notably, we demonstrate that the HM metabolome is dynamic during the period of maturity.

The human milk microbiome aligns with lactation stage and not birth mode.

We analysed the human milk microbiome in a cohort of 80 lactating women and followed the dynamics in taxa over the course of lactation from birth to 6 months. Two hundred and thirty one milk samples were collected from full-term lactating women at 1, 4, 8 and 24 weeks following birth and analysed for microbiota composition using 16S rRNA sequencing. A significant decrease in milk microbiota diversity was observed throughout the first 6 months of lactation, with the greatest difference seen between week 8 and week 24. Nine genera predominated in milk over lactation from week 1 to week 24, comprising of Staphylococcus, Streptococcus, Pseudomonas, Acinetobacter, Bifidobacterium, Mesorhizobium, Brevundimonas, Flavobacterium, and Rhodococcus; however, fluctuations in these core genera were apparent over time. There was a significant effect of stage of lactation on the microbiome, while no effect of birth mode, infant sex and maternal BMI was observed throughout lactation. Streptococcus had the highest mean relative abundance at week 1 and 24 (17.3% and 24% respectively), whereas Pseudomonas predominated at week 4 (22%) and week 8 (19%). Bifidobacterium and Lactobacillus had the highest mean relative abundance at week 4 (5% and 1.4% respectively), and occurred at a relative abundance of ≤ 1% at all other time points. A decrease in milk microbiota diversity throughout lactation was also observed. This study concluded that lactation stage was the primary driving factor in milk microbiota compositional changes over lactation from birth to 6 months, while mode of delivery was not a factor driving compositional changes throughout human lactation.

Interactions between whey proteins and calcium salts and implications for the formulation of dairy protein-based nutritional beverage products: A review.

Whey-based nutritional beverages are often fortified with calcium (Ca) in order to deliver the recommended intake of Ca. However, technical and product quality challenges are often experienced with Ca fortification of whey protein-based nutritional solutions, such as poor heat stability, high viscosity, colloidal instability, and impaired heat transfer. Understanding of the relationships and interactions between whey proteins and Ca relative to liquid process (e.g., ready to feed products, feed material prior to drying) is essential to designing and formulating nutritional whey-based products with desired physicochemical and colloidal stability properties. This article reviews the interactions between whey proteins and Ca salts used in the formulation of nutritional whey-based products as well as major processing implications associated with Ca fortification of whey-based solutions.

The impact of pH on mechanical properties, storage stability and digestion of alginate-based and soy protein isolate-stabilized emulsion gel beads with encapsulated lycopene.

In alginate-based emulsion gels containing protein-coated droplets, pH can influence the gelation mechanism of alginate gels, and the interactions between alginate molecules and protein-coated droplets, and thus properties of emulsion gels. This study investigated the impact of pH 3-7 on the properties (e.g., surface structures of droplets, mechanical properties, storage stability, digestion behavior) of alginate gel beads containing soy protein isolate(SPI)-stabilized oil droplets. Emulsion droplets were SPI-coated droplets at pH 6-7 and alginate/SPI-coated droplets at pH 3-5. Emulsion droplet flocculation only occurred in emulsions at pH 7.0. Emulsion gel beads at pH 3.0 had lower mechanical strength, higher storage stability, faster release of encapsulated lycopene during in-vitro digestion, and higher bioaccesibility of lycopene after 2 h of intestinal digestion than those at pH 7.0 and 5.0. The findings of this study are crucial to emulsion gel beads with controlled release and improved storage stability of encapsulated compounds by changing the pH of emulsions.

Effect of Protein Genotypes on Physicochemical Properties and Protein Functionality of Bovine Milk: A Review.

Milk protein comprises caseins (CNs) and whey proteins, each of which has different genetic variants. Several studies have reported the frequencies of these genetic variants and the effects of variants on milk physicochemical properties and functionality. For example, the C variant and the BC haplotype of αS1-casein (αS1-CN), ß-casein (ß-CN) B and A1 variants, and κ-casein (κ-CN) B variant, are favourable for rennet coagulation, as well as the B variant of ß-lactoglobulin (ß-lg). κ-CN is reported to be the only protein influencing acid gel formation, with the AA variant contributing to a firmer acid curd. For heat stability, κ-CN B variant improves the heat resistance of milk at natural pH, and the order of heat stability between phenotypes is BB > AB > AA. The A2 variant of ß-CN is more efficient in emulsion formation, but the emulsion stability is lower than the A1 and B variants. Foaming properties of milk with ß-lg variant B are better than A, but the differences between ß-CN A1 and A2 variants are controversial. Genetic variants of milk proteins also influence milk yield, composition, quality and processability; thus, study of such relationships offers guidance for the selection of targeted genetic variants.

Cold Microfiltration as an Enabler of Sustainable Dairy Protein Ingredient Innovation.

Classically, microfiltration (0.1-0.5 µm) of bovine skim milk is performed at warm temperatures (45-55 °C), to produce micellar casein and milk-derived whey protein ingredients. Microfiltration at these temperatures is associated with high initial permeate flux and allows for the retention of the casein fraction, resulting in a whey protein fraction of high purity. Increasingly, however, the microfiltration of skim milk and other dairy streams at low temperatures (≤20 °C) is being used in the dairy industry. The trend towards cold filtration has arisen due to associated benefits of improved microbial quality and reduced fouling, allowing for extended processing times, improved product quality and opportunities for more sustainable processing. Performing microfiltration of skim milk at low temperatures also alters the protein profile and mineral composition of the resulting processing streams, allowing for the generation of new ingredients. However, the use of low processing temperatures is associated with high mechanical energy consumption to compensate for the increased viscosity, and thermal energy consumption for inline cooling, impacting the sustainability of the process. This review will examine the differences between warm and cold microfiltration in terms of membrane performance, partitioning of bovine milk constituents, microbial growth, ingredient innovation and process sustainability.

Compositional and functional properties of milk and dairy products derived from cows fed pasture or concentrate-based diets.

Worldwide milk production is predominantly founded on indoor, high-concentrate feeding systems, whereas pasture-based feeding systems are most common in New Zealand and Ireland but have received greater attention recently in countries utilizing conventional systems. Consumer interest in 'pasture-fed' dairy products has also increased, arising from environmental, ethical, and nutritional concerns. A substantial body of research exists describing the effect of different feeding strategies on the composition of milk, with several recent studies focusing on the comparison of pasture- and concentrate-based feeding regimes. Significant variation is typically observed in the gross composition of milk produced from different supplemental feeds, but various changes in the discrete composition of macromolecular components in milk have also been associated with dietary influence, particularly in relation to the fatty acid profile. Changes in milk composition have also been shown to have implications for milk and dairy product processability, functionality and sensory properties. Methods to determine the traceability of dairy products or verify marketing claims such as 'pasture-fed' have also been established, based on compositional variation due to diet. This review explores the effects of feed types on milk composition and quality, along with the ultimate effect of diet-induced changes on milk and dairy product functionality, with particular emphasis placed on pasture- and concentrate-based feeding systems.

A Proteomic Study for the Discovery of Beef Tenderness Biomarkers and Prediction of Warner-Bratzler Shear Force Measured on Longissimus thoracis Muscles of Young Limousin-Sired Bulls.

Beef tenderness is of central importance in determining consumers' overall liking. To better understand the underlying mechanisms of tenderness and be able to predict it, this study aimed to apply a proteomics approach on the Longissimus thoracis (LT) muscle of young Limousin-sired bulls to identify candidate protein biomarkers. A total of 34 proteins showed differential abundance between the tender and tough groups. These proteins belong to biological pathways related to muscle structure, energy metabolism, heat shock proteins, response to oxidative stress, and apoptosis. Twenty-three putative protein biomarkers or their isoforms had previously been identified as beef tenderness biomarkers, while eleven were novel. Using regression analysis to predict shear force values, MYOZ3 (Myozenin 3), BIN1 (Bridging Integrator-1), and OGN (Mimecan) were the major proteins retained in the regression model, together explaining 79% of the variability. The results of this study confirmed the existing knowledge but also offered new insights enriching the previous biomarkers of tenderness proposed for Longissimus muscle.

Shotgun proteomics for the preliminary identification of biomarkers of beef sensory tenderness, juiciness and chewiness from plasma and muscle of young Limousin-sired bulls.

Label free shotgun proteomics was used to analyse plasma and Longissimus muscle biopsies of Limousin-sired bulls, classified as 5 high-quality and 5 low-quality meat based on sensory texture traits (tenderness, juiciness and chewiness). A total of 31 putative protein biomarkers (16 in plasma and 15 in muscle) differed significantly in abundance between the two quality groups. The proteins were associated with muscle structure, energy metabolism, heat shock proteins, oxidative stress and proteolysis related pathways. Among them, B2M, AHSG, APOA4 and HP-20 (plasma), PFKM, MYH2, PTER, GSTM1 and MYPN (muscle) were good predictors of the three texture quality traits. Further, significant correlations were identified for FETUB, SERPINA7, ASL, TREH, HP, HP-25, AZGP1, APCS and SYT15, which are novel biomarkers from plasma that warrant further evaluation. This study is a significant step forward in elucidating proteomic profiles in bovine bio-fluids and muscle tissue, which may ultimately provide opportunities to processors for early assessment of beef sensory quality.

Protein levels and protease activity in milk from mothers of pre-term infants: A prospective longitudinal study of human milk macronutrient composition.

BACKGROUND AND AIMS: The composition and enzymology of human milk changes throughout the lactation period, and differ for mothers who give birth prematurely compared to those who deliver at full-term. Understanding the composition of milk from mothers of very low birth weight premature infants is of great significance, and the objective of this study was to evaluate the composition, protein profile and plasmin activity of milk from mothers who delivered infants at different gestational ages. METHODS: Samples of human milk were donated by women (n = 74) in the Cork, Ireland, area who gave birth to full-term (>37 weeks gestation, FT), pre-term (32-37 weeks, PT) and very pre-term (≤32 weeks, VPT) infants. FT milk was collected at 1, 3, 6 and 10 weeks post-partum (PP), while PT and VPT milk was collected weekly until the FT due date of the infant and subsequently followed the FT protocol. RESULTS: Gestational age did not significantly affect lactose or fat content or total energy content of milk. However, protein content, and levels of some individual proteins, were significantly affected by both gestational age at birth and duration of lactation, with significantly higher protein levels in PT or VPT milk samples at 0-7 days and 1-2 months, respectively. Plasmin activity was significantly higher in VPT milk, indicating differences in proteolytic processing in milk. CONCLUSION: Compositional differences between the milk of mothers of term and pre-term infants were greatest in terms of the protein profile, which showed both qualitative and quantitative differences, as well as difference in proteolytic activity.

Solubility of carbon dioxide in renneted casein matrices: Effect of pH, salt, temperature, partial pressure, and moisture to protein ratio.

The solubility of carbon dioxide (CO2) in the moisture and protein components of cheese matrices and the influence of changing pH, salt and temperature levels remains unclear. In this study, model casein matrices were prepared, by renneting of micellar casein concentrate (MCC), with modulation of salt and pH levels by adding salt and glucono delta-lactone, respectively, to the MCC solutions prior to renneting. Different moisture-to-protein levels were achieved by freeze-drying, incubation of samples at different relative humidities, or by applying varying pressures during gel manufacture. The CO2 solubility of samples decreased linearly with both increasing temperature and salt-in-moisture content, whereas solubility of CO2 increased with increasing pH. A non-linear relationship was observed between CO2 solubility and the moisture-to-protein ratio of experimental samples. Overall, such knowledge may be applied to improve the quality and consistency of eye-type cheese, and in particular to avoid development of undesirable slits and cracks.

Colorants in cheese manufacture: Production, chemistry, interactions, and regulation.

Colored Cheddar cheeses are prepared by adding an aqueous annatto extract (norbixin) to cheese milk; however, a considerable proportion (∼20%) of such colorant is transferred to whey, which can limit the end use applications of whey products. Different geographical regions have adopted various strategies for handling whey derived from colored cheeses production. For example, in the United States, whey products are treated with oxidizing agents such as hydrogen peroxide and benzoyl peroxide to obtain white and colorless spray-dried products; however, chemical bleaching of whey is prohibited in Europe and China. Fundamental studies have focused on understanding the interactions between colorants molecules and various components of cheese. In addition, the selective delivery of colorants to the cheese curd through approaches such as encapsulated norbixin and microcapsules of bixin or use of alternative colorants, including fat-soluble/emulsified versions of annatto or beta-carotene, has been studied. This review provides a critical analysis of pertinent scientific and patent literature pertaining to colorant delivery in cheese and various types of colorant products on the market for cheese manufacture, and also considers interactions between colorant molecules and cheese components; various strategies for elimination of color transfer to whey during cheese manufacture are also discussed.

The Effect of Compositional Changes Due to Seasonal Variation on Milk Density and the Determination of Season-Based Density Conversion Factors for Use in the Dairy Industry.

The objective of this study was to determine the effect of seasonal variation on milk composition and establish an algorithm to predict density based on milk composition to enable the calculation of season-based density conversion calculations. A total of 1035 raw whole milk samples were collected from morning and evening milking of 60 spring-calving individual cows of different genetic groups, namely Jersey, Elite HF (Holstein-Friesian) and National Average HF, once every two weeks for a period of 9 months (March-November, 2018). The average mean and standard deviation for milk compositional traits were 4.72 ± 1.30% fat, 3.85 ± 0.61% protein and 4.69 ± 0.30% lactose and density was estimated at 1.0308 ± 0.002 g/cm3. The density of the milk samples was evaluated using three methods: a portable density meter, DMA 35; a standard desktop version, DMA 4500M; and an Association of Official Agricultural Chemists (AOAC) method using 100-mL glass pycnometers. Statistical analysis using a linear mixed model showed a significant difference in density of milk samples (p < 0.05) across seasonal and compositional variations adjusted for the effects of days in milk, parity, the feeding treatment, the genetic group and the measurement technique. The mean density values and standard error of mean estimated for milk samples in each season, i.e., spring, summer and autumn were 1.0304 ± 0.00008 g/cm3, 1.0314 ± 0.00005 g/cm3 and 1.0309 ± 0.00007 g/cm3, respectively.

Physical and colloidal stability of conventional and micronised calcium citrate ingredient powders in the formulation of infant nutritional products.

The fortification of food systems with calcium remains difficult; one challenge is to maintain the colloidal stability of insoluble calcium salts during processing and shelf life. Particle size reduction of insoluble salts may result in improved product stability. In this study, insoluble calcium citrate in two different particle sizes, conventional calcium citrate and micronised calcium citrate, were first evaluated in terms of physical and bulk handling properties, followed by protein adsorption and colloidal stability when dispersed in two dairy-based nutritional beverages differing in composition, i.e. infant milk formula stage 1 and 3. Particle size distribution analysis showed micronised calcium citrate (volume-weighted diameter = 5.10 µm) to have significantly smaller (p < 0.05) particle size than conventional calcium citrate (volume-weighted diameter = 88.2 µm). The adsorption of dairy proteins onto particles of calcium citrate resulted in caseins having greater affinity for both salts, followed by ß-lactoglobulin. The smaller particle size of the micronised citrate resulted in higher affinity for casein and greater colloidal stability when dispersed in both infant milk formula solutions compared to conventional calcium salts. The results of this study provide knowledge on the application of micronised insoluble calcium salts in the fortification of nutritional dairy-based products.

Effect of Pseudomonas fluorescens proteases on the quality of Cheddar cheese.

The objective of this study was to investigate the effect of adding different levels of a thermoresistant protease produced by a Pseudomonas fluorescens strain to milk on the manufacture and quality of Cheddar cheese. Fresh raw milk was collected, standardized, and pasteurized at 72°C for 15 s, and the enzyme was added to give a protease activity of 0.15 or 0.60 U/L (treatments P1 and P4, respectively), while one sample had no enzyme added (control). Milk was stored at 4°C for 48 h and Cheddar cheese was manufactured after 0 and 48 h of storage. Results indicated that the protease was active in milk during 48 h of storage; however, its effect on milk composition was minimal. The protein that was preferentially hydrolyzed by the protease over storage was ß-casein, followed by κ-casein. The mean cheese yield and recovery of fat and protein obtained for all cheeses were not affected by protease activity. The protease showed low activity during cheese manufacture, possibly because of unfavorable conditions, including low pH. One of the factors that might have influenced protease activity was the pH of the curd (approximately 6.55 after acidification and 5.35 at milling), which was lower than that at which the enzyme would have optimum activity (pH 7 to 9). Consequently, the composition, pH, patterns of proteolysis, and hardness of all cheeses produced were similar and in accordance with values expected for that type of cheese, independently of the protease activity level. However, slight increases in proteolysis were observed in P4 cheeses and produced using milk stored for 48 h. Both the P1 and P4 cheeses had higher concentrations of free amino acids (FAA) compared with the control, whereas urea-PAGE electrophoretograms indicated a greater breakdown of caseins in the P4 cheese samples, which may be related to possible increases in numbers of proteolytic bacteria in milk during storage. Therefore, the thermoresistant psychrotrophic bacterial protease(s) tested in this study may affect the manufacture or quality of Cheddar cheese during ripening to a relatively limited extent. However, controlling initial levels of proteolytic bacteria in raw milk remains essential, because proteolysis affects the development of flavor and texture in cheese.

Effect of Diet on the Vitamin B Profile of Bovine Milk-Based Protein Ingredients.

The influence of diet on the water-soluble vitamin composition of skim milk powder and whey protein ingredients produced from the milk of cows fed pasture or concentrate-based diets was examined. Fifty-one Holstein-Friesian cows were randomly assigned into three diets (n=17) consisting of outdoor grazing of perennial ryegrass (GRS), perennial ryegrass/white clover (CLV), or indoor feeding of total mixed ration (TMR) for an entire lactation. Raw mid-lactation milk from each group was processed into skim milk powder and further processed to yield micellar casein whey and acid whey. Sweet whey was also produced by renneting of pasteurised whole milk from each system. The water-soluble vitamin profile of each sample was analysed using a combination of direct injection mass spectrometry and reverse-phase liquid chromatography-mass spectrometry. Vitamin B3 and B3-amide concentrations were significantly higher (p < 0.05) in TMR-derived samples than in those from CLV and GRS, respectively. Vitamin B1, B2, and B7 concentrations were significantly higher in GRS and CLV-derived samples than those from TMR. Significant differences in vitamins B1, B2, and B3-amide were also observed between protein ingredient types. This study indicates that bovine feeding systems have a significant effect on B vitamin composition across a range of protein ingredient types.