Date of Award


Degree Name

MS in Agriculture - Dairy Products Technology


Dairy Science


Phillip S. Tong


The proteins found in milk are largely important in the functionality of many dairy products and dairy processes. The casein micelle system in milk is a complex and highly studied system. The micelle is thought to be a sponge like structure containing four caseins, αs1, αs2, β, and κ casein, and bound together with colloidal calcium phosphate. When a chelating agent such as a citrate, phosphate, or polyphosphate are added to milk systems, the CCP is bound to the chelator and removed from the micelle. It has been shown through past research that the use of calcium chelating agents disrupts the calcium phosphate equilibrium and allows for the dissociation of the casein micelle and release of the individual caseins. Once the caseins are disrupted from micellar form and in solution, it may be possible to separate out different casein streams for functional usage in dairy products using common separation techniques.

This thesis project seeks to evaluate the feasibility of separating milk treated with calcium chelators using various separation techniques to evaluate the individual casein fractions of this disrupted system. Four separation methods (ultracentrifugation, membrane filtration, heat coagulation, and coagulation based on pH) were employed to separate out the caseins based on selected properties, specifically density, molecular weight, and solubility. In ultracentrifugation, three speeds were tested, the heat coagulation study tested two temperatures, and pH based coagulation tested four different pHs to determine their impact on overall protein levels and individual casein yields. Skim milk powder was reconstituted and chelator was added at 1, 50, or 100 mEq/L treatment level. These samples were then separated using aforementioned techniques, and the supernatant or permeate was analyzed for total protein content, individual casein composition, turbidity, and mean particle size.

Analysis of centrifugal separation studies shows the interaction between chelator type, chelator level, and centrifugation speed had a significant impact on the amount of protein released from the casein micelle (p

Coagulation trials based on pH were also shown to have a significant interaction between chelator type, chelator level, and sample pH effecting the protein levels and casein composition (p

Membrane filtration showed low protein yields in permeate, however trisodium citrate 100 mEq was still shown to have significantly higher permeate % protein levels (p

The use of heat based coagulation as an individual casein separation technique for chelated samples is not recommended, as the casein micelle system itself is extremely heat stable, and the use of calcium chelators only increases the heat stability further. Because of the increased heat stability, no coagulum was formed in samples upon heating, and therefore, no separation and analysis could be done.

Improving our knowledge of pretreatment of milk prior to separation and the effectiveness of different separation methods on chelated milk products may result in information leading to the ability to separate out milk fractions that provide unique or improved properties for product applications.

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Dairy Science Commons