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Slide 2 - Functional Properties of Proteins 3 major categories Hydration properties Protein to water interactions Dispersibility, solubility, adhesion, Water holding capacity, viscosity Structure formation Protein to protein interactions Gel formation, precipitation, Aggregation Surface properties Protein to interface interactions Foaming, emulsification
Slide 3 - Coconut Water-Milk Milk protein concentrate (90% protein) Coconut water concentrate (rich in Na, K, Mg, Ca) Phosphate buffers to pH 7.0 Gum acacia Locust bean gum Flavors, minor ingredients
Slide 4 - Coconut Water-Milk Milk protein concentrate (90% protein) Processed by ultra-filtration Passed through a membrane to remove water Washed over and over to remove: Some whey proteins Minerals Lactose Water
Slide 5 - ppt slide no 5 content not found
Slide 6 - Coconut Water-Milk The Process: Buffers added to 35°C water (pH 11) MPC added, mixed for 30 min with shear Gums added Coconut water added Homogenized Aseptically processed (140°C for 15 sec) Homogenized again Packaged cold
Slide 7 - Coconut Water-Milk The Issue: Product has 20 gram of protein per 325 mL serving Very thick, rich, and creamy After 6 months on-shelf, it begins to fade. Mild clumping of proteins Sediment formation Larger protein clumps form Looks like curdled milk Flavor is fine, no detectable microorganisms
Slide 8 - Coconut Water-Milk Initial diagnosis: Protein oxidation? Too high of a mineral content Vitamin and mineral supplement was removed Whey proteins were denaturing, loosing water Slow and steady drop in pH was observed pH 7.0 to pH 6.2 over 6 months Need to re-buffer the system
Slide 9 - Coconut Water-Milk First attempts: Add antioxidants (ascorbic, sulfite, polyphenolics) Aded oxidizers (cross-link proteins) Add more/stronger/different buffers Adding more gum to stabilize the water Slow down the shear rate Reduce foaming to prevent protein denaturation Adding oils to help stabilize protein hydration
Slide 10 - Coconut Water-Milk Nothing worked. In lab trails, the product was form precipitates in 2-3 days at 37°C, compared to 6 months in commercial product. Almost impossible to simulate the industrial process of homogenization and UHT processing and re-homogenization pH drops from 7.0 to 6.0 in 24-hrs Drops to <5.0 in 48 hrs
Slide 11 - Coconut Water-Milk So why a decrease in pH? A natural ion exchange is taking place Milk caseins are mostly negatively charged Phosphates are negatively charged CWC is mostly cationic, and slighly acidic Where does the H+ come from? Most likely….from the casein and phosphates Exchanging ions with cationic minerals
Slide 12 - Coconut Water-Milk
Slide 13 - Coconut Water-Milk
Slide 14 - Coconut Water-Milk
Slide 15 - Coconut Water-Milk So here is one of the craziest reactions I have ever seen. Adding an ALKALI and a resultant LOWERING of the pH Add CWC to alkaline phosphate buffer (with or without MPC) to hit pH 7.0. Allow it to hold at 37°C, and the pH slowly drops with time (ion exchange) Add MORE alkaline phosphate to raise the pH back to pH 7 Over minutes to hours, the pH will lover quickly to pH <6. You can bounce it like a yo-yo….pH high, pH low, pH high, pH low
Slide 16 - Coconut Water-Milk What did work: Changing the order of ingredient addition Blending the phosphates (negatively charged, alkaline phosphates) with the CWC causes an immediate buffering with the ions present So this SLOWS down the decrease in pH But does not completely stop the precipitation pH eventually drops, and proteins fall out
Slide 17 - Coconut Water-Milk What were the eventual solutions? #1. Change the order of ingredients, to allow chelation of CWC minerals with phosphates #2. Stop the ion-exchange by physical blocking of the reaction with ionic gums
Slide 18 - Coconut Water-Milk At it also turns out, gum hydration temperatures were too low. 35°C is insufficient to hydrate gum acacia, it needs temps closer to 70°C Gum acacia is an ionic gum, so it is participating in the ion exchange reactions. With the gum poorly hydrated, it was binding to water OUTSIDE the milk protein. Hardest problems often have simplest solutions Increased the processing temperature, problem gone!