Crystallization of Lactose-Protein Solutions in the Presence of Flavonoids.

Lactose is commonly crystallized in the presence of whey proteins, forming co-crystals of lactose and proteins. This work hypothesized that flavonoids such as rutin or epigallocatechin-3-gallate (EGCG) could be incorporated into the lactose and protein co-crystal structure since flavonoids may interact with both lactose and proteins. The interactions between whey proteins and flavonoids were first studied. Then, lactose-protein solutions were crystallized with and without flavonoids, measuring the kinetic parameters of crystallization and characterizing the resulting crystals. The incorporation of flavonoids in lactose-protein co-crystals depended on the hydrophilic nature of flavonoids. The hydrophilic EGCG was scarcely enclosed in the crystal lattice of lactose and avoided the inclusion of whey proteins in the crystals. In contrast, the less water-soluble rutin interacted with whey proteins and lactose, leading to the formation of co-crystals containing lactose, protein, and a large concentration of rutin (3.468 ± 0.392 mg per 100 mg of crystals).

Mutarotation and solubility of lactose as affected by carrageenans.

It has been reported that polysaccharides like carrageenan can change the crystallization of lactose. However, it is still unclear whether changes in lactose mutarotation, solubility, and super-solubility are involved in carrageenans' effect on lactose crystallization. It has been established that the conversion of α- to ß-lactose forms (mutarotation) in an aqueous solution has a significant impact on lactose crystallization. Similarly, lactose solubility changes lead to changes in the metastable zone (MZ), a region between the solubility and super-solubility of lactose. The width of this MZ determines the temperature drop necessary to induce lactose nucleation. This work aimed to study the effect of carrageenans on lactose mutarotation and solubility. For this purpose, lactose solutions were added with ι and κ- carrageenan at two concentrations: 50 and 100 mg L-1. Optical rotation measurements estimated the proportion of ß/α isomers in lactose solutions. Besides, solubility and super- solubility was determined to build the MZ. The presence of carrageenans changed both the time to reach the mutarotation balance and the proportion of ß/α isomers at mutarotation equilibrium. Carrageenans decreased the solubility of lactose in a range of temperatures between 10 and 60 °C and reduced the metastable zone width (MZW).

Individual and combined effect of pH and whey proteins on lactose crystallization.

Lactose is recovered by crystallization from cheese whey that is a by-product of cheesemaking. The whey used for the recovery of lactose usually has a residual content of protein that alters the crystallization of lactose. In addition, the pH of whey may fluctuate depending on the cheese variety. However, there is little information on how the pH modifies the effect that whey proteins have on lactose crystallization. Accordingly, this work aimed to evaluate the individual and combined effect of whey proteins and pH on the kinetics of crystallization, the crystal size distribution and the crystallinity of lactose. The addition of whey proteins in lactose solutions (25% v/v) modified the process of lactose crystallization. However, the effect that whey proteins had on lactose crystallization heavily depended on the pH. The number of crystals per milliliter as well as the growth and size distribution of crystals was the most affected with the changes in pH (pHs of 7, 5.5 and 4) and the addition of whey proteins (0 and 0.63%). All the treatment produced mostly α-lactose monohydrated but some treatments also generated crystals of ß-lactose (pH 5.5, 0% of proteins). Amorphous lactose was observed mainly in lactose solutions adjusted at pH 7 and added with whey proteins. This particular treatment also incorporated the highest amount of protein into the lattice of lactose crystals. The results of this work highlight the importance of controlling the pH of lactose crystallization, especially if there is a presence of whey proteins.

Ultrasound-assisted crystallization of lactose in the presence of whey proteins and κ-carrageenan.

The conventional process of lactose crystallization is prolonged, hardly controllable and the crystals have low quality. In this work, the effect of ultrasound on the crystallization of lactose in an aqueous system was assessed. Additionally, it was studied how the presence of whey proteins (which are a common impurity) and κ-carrageenan (that possess high water-binding capacity) could modify the process of lactose crystallization. Lactose solutions at 25% were sonicated in a continuous flow chamber at two different energy densities (9 and 50 J mL-1) before the start of crystallization. Some of these lactose solutions were previously added with κ-carrageenan (0, 150 and 300 mg L-1), with whey proteins (0.64%) or with both at the same time. Ultrasound sped up the rate of crystallization, decreased the crystal's size and narrowed the crystal size distribution (CSD). The presence of whey proteins accelerated the process of crystallization but induced the formation of amorphous lactose. Likewise, the rate of lactose crystallization was improved by the addition of 150 mg L-1 of carrageenan. Whereas, the combination of carrageenan and whey proteins generated the smallest crystals (6 µm), the narrowest CSD and minimized the formation of amorphous lactose.