Effects of different calcium salts on properties of milk related to heat stability

Insoluble calcium salts were added to milk to increase total calcium by 30 mM, without changing properties influencing heat stability, such as pH and ionic calcium. There were no major signs of instability associated with coagulation, sediment formation or fouling when subjected to ultra high temperature (UHT) and in‐container sterilisation. The buffering capacity was also unaltered. On the other hand, addition of soluble calcium salts reduced pH, increased ionic calcium and caused coagulation to occur. Calcium chloride showed the largest destabilising effect, followed by calcium lactate and calcium gluconate. Milk became unstable to UHT processing at lower calcium additions compared to in‐container sterilisation.     

The measurement and significance of ionic calcium in milk – A review

The measurement of ionic calcium in milk and milk products is of vital importance in understanding the role of calcium in milk. This review compares the methods of measurement including ion equilibration, murexide and ion selective electrodes. Secondly the variations in milk from individual cows and goats, and in bulk milk samples are reviewed. The third section examines the differences in ionic calcium in relation to processing: addition and removal of calcium and other salts, rennet coagulation, miscellaneous influences, filtration and evaporation and drying are all reviewed. Finally the review considers why ionic calcium measurement is not more widely measured within the diary industry and argues that it could be beneficial to do so.     

Natural variations of citrate and calcium in milk and their effects on milk processing properties

Natural variations among milk constituents, and their relations to each other as well as to processing parameters, represent possibilities for differentiation of milk to produce high-quality natural products. In this study, we focused on natural variations in milk citrate and its interplay with calcium distribution in milk, in relation to processing properties. Milk samples from individual cows from farms varying in feeding and management practices were collected from April to June 2017 to maximize natural variations in citrate and calcium. Chemical composition, rennet coagulation properties, and ethanol stability were analyzed for all milk samples. We focused particularly on calcium distribution and citrate content and the correlation of these to other milk parameters. No significant change in citrate content was observed during the sampling period, which suggests that mechanisms other than feeding affect citrate levels in milk. Several significant correlations were found, including a positive correlation between complexed serum calcium and citrate, and a negative correlation between urea and ionic calcium. These are both of interest in relation to further processing, as with regard to the stability of UHT milk and in cheese making. Although the correlation between complexed serum calcium and citrate may be explained by their affinity, the underlying driver for the negative relationship between natural milk urea and ionic calcium needs to be clarified by further studies. Furthermore, milk from the different farms varied not only with regard to organic versus conventional farming systems; feeding practices between farms also play an important role in milk composition and functionality. However, none of the differences in milk composition between farms were found to decrease milk functionality and thus would probably not cause any processing problems.     

Measurement of ionic calcium in milk

There are many reports in the literature regarding the effects of ionic calcium on reactions related to casein micelle stability, such as heat stability, ethanol stability and susceptibility to gelation, sediment formation and fouling. However, experimental evidence supporting these assertions is much less readily available.
This paper evaluates three selective ion electrode systems for measuring ionic calcium directly in milk as well as looking at the effects on pH reduction and addition of calcium chloride.
The best electrode system was the Ciba Corning 634 system, which was designed for blood but has been modified for milk. This was found to be reproducible and stable when calibrated daily and allowed direct measurements to be taken on milk in 70 s. This has been found to perform well now for 3 years. The other systems were not so useful, as they took longer to stabilize, but may be useful for higher ionic calcium concentrations, which are found in acidified milk products.
Reducing the pH increased ionic calcium and reduced ethanol stability. Calcium chloride addition reduced pH, increased ionic calcium and reduced the ethanol stability. Readjusting the pH to its value before calcium addition reduced the ionic calcium, but not back to its original value. Milks from individual cows showed wide variations in their ionic calcium concentrations.
This establishes the methodology for a more detailed investigation on measurement of ionic calcium in milks from individual cows and from bulk milks, to allow a better understanding of its role in casein micelle stability.     

Effects of stabiliser addition and in-container sterilisation on selected properties of milk related to casein micelle stability

Different stabilising salts and calcium chloride were added to raw milk to evaluate changes in pH, ionic calcium, ethanol stability, casein micelle size and zeta potential. These milk samples were then sterilised at 121 °C for 15 min and stored for 6 months to determine how these properties changed. Addition of tri-sodium citrate (TSC) and di-sodium hydrogen phosphate (DSHP) to milk reduced ionic calcium, increased pH and increased ethanol stability in a concentration-dependent fashion. There was relatively little change in casein micelle size and a slight decrease in zeta potential. Sodium hexametaphosphate (SHMP) also reduced ionic calcium considerably, but its effect on pH was less noticeable. In contrast, sodium dihydrogen phosphate (SDHP) reduced pH but had little effect on ionic calcium. In-container sterilisation of these samples reduced pH, increased ethanol stability and increased casein micelle size, but had variable effects on ionic calcium; for DSHP and SDHP, ionic calcium decreased after sterilisation but, for SHMP, it remained little changed or increased. Milk containing 3.2 mM SHMP and more than 4.5 mM CaCl₂ coagulated upon sterilisation. All other samples were stable but there were differences in browning, which increased in intensity as milk pH increased. Heat-induced sediment was not directly related to ionic calcium concentration, so reducing ionic calcium was not the only consideration in terms of improving heat stability. After 6 months of storage, the most acceptable product, in appearance, was that containing SDHP, as this minimised browning during sterilisation and further development of browning during storage.     

Measurement of ionic calcium, pH, and soluble divalent cations in milk at high temperature

Dialysis and ultrafiltration were investigated as methods for measuring pH and ionic calcium and partitioning of divalent cations of milk at high temperatures. It was found that ionic calcium, pH, and total soluble divalent cations decreased as temperature increased between 20 and 80°C in both dialysates and ultrafiltration permeates. Between 90 and 110°C, ionic calcium and pH in dialysates continued to decrease as temperature increased, and the relationship between ionic calcium and temperature was linear. The permeabilities of hydrogen and calcium ions through the dialysis tubing were not changed after the tubing was sterilized for 1 h at 120°C. There were no significant differences in pH and ionic calcium between dialysates from raw milk and those from a range of heat-treated milks. The effects of calcium chloride addition on pH and ionic calcium were measured in milk at 20°C and in dialysates collected at 110°C. Heat coagulation at 110°C occurred with addition of calcium chloride at 5.4 mM, where pH and ionic calcium of the dialysate were 6.00 and 0.43 mM, respectively. Corresponding values at 20°C were pH 6.66 and 2.10 mM.     

Influence of calcium salt supplementation on calcium equilibrium in skim milk during pH cycle

Calcium is a mineral essential for humans, especially for bone constitution. Yet most of the worldwide population does not satisfy their Ca needs. Hence, Ca supplementation is of major importance, even in western countries where some specific populations at risk do not satisfy the recommended daily intake of Ca. More than 70% of dietary Ca comes from dairy products. Calcium supplementation of naturally Ca-rich sources such as skim milk is then of special interest. To our knowledge, few data are available concerning milk Ca (MC) supplementation of milk, particularly when followed by pH cycle. In this paper, MC supplementation is studied and compared with Ca chloride (CC) supplementation as a well-known source of Ca. The effect of Ca salt supplementation followed by pH cycle was studied in reconstituted skim milk. Calcium supplementation was carried out with CC and MC at 25 mmol of Ca/kg of skim milk. Ionized Ca concentration and turbidity variations were followed in situ by Ca ion selective electrode and turbidimetry using light reflection. From normalized data on ionized Ca concentration and turbidity vs. pH, it appeared that hysteresis areas were smaller for CC-supplemented milk, whereas unsupplemented milk and MC-supplemented milk behaved similarly. For these 3 dairy systems, pH cycles to pH 5.0 led to a larger hysteresis area than pH cycles to pH 5.5. The shrinkage of the hysteresis area could be interpreted as a reinforcement of casein micelles with Ca ions over the pH cycle.     

Effect of pH and calcium concentration on some textural and functional properties of mozzarella cheese

The effects of Ca concentration and pH on the composition, microstructural, and functional properties of Mozzarella cheese were studied. Cheeses were made using a starter culture (control) or by direct acidification of the milk with lactic acid or lactic acid and glucono-delta-lactone. In each of three trials, four cheeses were produced: a control, CL, and three directly-acidified cheeses, DA1, DA2, and DA3. The cheeses were stored at 4 degrees C for 70 d. The Ca content and pH were varied by altering the pH at setting, pitching, and plasticization. The mean pH at 1 d and the Ca content (mg/g of protein) of the various cheeses were: CL, 5.42 and 27.7; DA1, 5.96 and 21.8; DA2, 5.93 and 29.6; DA3, 5.58 and 28.7. For cheeses with a high pH (i.e., approximately 5.9), reducing the Ca content from 29.6 to 21.8 mg/g of protein resulted in a significant decrease in the protein level and increases in the moisture content and mean level of nonexpressible serum (g/g of protein). Reducing the Ca concentration also resulted in a more swollen, hydrated para-casein matrix at 1 d. The decrease in Ca content in the high-pH cheeses coincided with increases in the mean stretchability and flowability of the melted cheese over the 70-d storage period. The fluidity of the melted cheese also increased when the Ca content was reduced, as reflected by a lower elastic shear modulus and a higher value for the phase angle, delta, of the melted cheese, especially after storage for <12 d. The melt time, flowability, and stretchability of the low-Ca, high-pH DA1 cheese at 1 d were similar to those for the CL cheese after storage for > or = 12 d. In contrast, the mean values for flowability and stretchability of the high-pH, high-Ca DA2 cheese over the 70-d period were significantly lower than those of the CL cheese. Reducing the pH of high-Ca cheese (27.7 to 29.6 mg/g of protein) from -5.95 to 5.58 resulted in higher flowability, stretchability, and fluidity of the melted cheese. For cheeses with similar pH and Ca concentration, the method of acidification had little effect on composition, microstructure, flowability, stretchability, and fluidity of the melted cheese.     

The effects of high salt and low pH on the water-holding of meat

The study investigated the water-holding (WH) in meat in the pH–NaCl (ionic strength) combinations that prevail in dry sausages during fermentation and drying. WH in raw beef homogenates, with 230% added water, was determined by centrifugation at pH values of 5.47–4.60, and ionic strengths (μ) 0.50–1.50. The minimum WH in relation to pH was at pH 4.8, but at higher pH values, the WH optimum was at 1.0–1.5 μ; at lower pH-values (< 5.0) the optimum was more pronounced at 1.0 μ. The WH reducing effect by pH decrease was stronger than the effect of μ. At lower pH values, the relative effect of μ on WH was higher compared to that of pH than at higher pH values. The pH–salt combinations prevailing in fermented sausage in the beginning of the ripening produced a high WH, which decreased, first with pH decrease and then in the last period of ripening mainly due to the increase of ionic strength.     

Milk reversibility following reduction and restoration of pH

The pH of skim milk was adjusted from 6.67 to between 6.00 and 5.45 before being restored with NaOH. Ionic calcium was higher after pH restoration but this did not result in instability during indirect ultra high temperature and in‐container sterilisation. Ionic calcium of pH restored milk increased with increased holding time but the pH restored milk remained stable to in‐container sterilisation. The viscosity of the milk was not affected by pH reversal. However, pH‐restoration reduced rennet coagulation time but milk was still stable when subjected to high temperature treatment. Properties of the soluble phase were measured at high temperature by dialysis.     

High-pressure structuring of milk protein concentrate: Effect of pH and calcium

In this study, we investigated the effect of pH and calcium on the structural properties of gels created by high-pressure processing (HPP, 600 MPa, 5°C, 3 min) of milk protein concentrate (MPC, 12.5% protein). The pH level of the MPC was varied between 6.6 and 5.1 by adding glucono-δ-lactone (GDL), and the calcium content was varied from 24 to 36 mg of Ca/g of protein by adding calcium chloride. The rheological properties and microstructure of the pressure-treated MPC were assessed. The pressurization treatments and analytical testing were conducted in triplicate. Data were analyzed statistically using one-way ANOVA with Tukey's honestly significant difference post hoc tests. A pressurization time of 3 min was sufficient to induce gel formation in MPC at pH 6.6, so it was used throughout the study. Adjusting either pH or calcium affected the structure of the HPP-created milk protein gels, likely by influencing electrostatic interactions and shifting the calcium–phosphate balance. Gels were formed after pressurization of MPC at pH above 5.3, and increasing the pH from 5.3 to 6.6 resulted in stronger gels with higher values of elastic moduli (G′). At neutral pH (6.6), adding calcium to MPC further increased G′. Scanning electron microscopy showed that reducing pH or adding calcium resulted in more porous, aggregated microstructures. These findings demonstrate the potential of HPP to create a variety of structures using MPC, facilitating a new pathway from dairy protein ingredients to novel, gel-based, high-protein foods, such as puddings or on-the-go protein bars.     

Effect of pH and calcium concentration on proteolysis in mozzarella cheese

Low-moisture Mozzarella cheeses (LMMC), varying in calcium content and pH, were made using a starter culture (control; CL) or direct acidification (DA) with lactic acid or lactic acid and glucono-delta-lactone. The pH and calcium concentration significantly affected the type and extent of proteolysis in Mozzarella cheese during the 70-d storage period at 4 degrees C. For cheeses with a similar pH, reducing the calcium-to-casein ratio from -29 to 22 mg/g of protein resulted in marked increases in moisture content and in primary and secondary proteolysis, as indicated by polyacrylamide gel electrophoresis and higher levels of pH 4.6- and 5%-PTA-soluble N. Increasing the pH of DA cheeses of similar moisture content, from approximately 5.5 to 5.9, while maintaining the calcium-to-casein ratio almost constant at approximately 29 mg/g, resulted in a decrease in primary proteolysis but had no effect on secondary proteolysis. Comparison of CL and DA cheeses with a similar composition showed that the CL cheese had higher levels of alpha(s1)-CN degradation, pH 4.6- and 5%-PTA-soluble N. Analysis of pH 4.6-soluble N extracts by reverse-phase HPLC showed that the CL cheese had higher concentrations of compounds with low retention times, suggesting higher concentrations of low molecular mass peptides and free amino acids.     

Bioavailability of calcium and physicochemical properties of calcium-fortified buffalo milk

Buffalo milk was fortified with calcium at the rate of 50 mg calcium/100 ml milk using calcium chloride, calcium lactate and calcium gluconate, and the resulting decrease in pH was restored to its original value by adding disodium phosphate. The maximum heat stability of calcium-fortified buffalo milk remained slightly lower than that of unfortified milk. Calcium gluconate-fortified milk had the highest heat stability, bioavailability of calcium, partitioning of calcium in the dissolved state and viscosity, and the least curd tension compared to other fortified milk, without any adverse impact on sensory properties. The bioavailability of calcium and heat stability was lowest in the case of buffalo milk fortified with calcium chloride.     

中性高钙液体奶的钙剂筛选及稳定性研究

对中性高钙液体奶生产中不同钙剂和稳定剂对产品稳定性和品质的影响进行了研究。结果表明,以乳钙为钙强化剂,添加量为6g/L,微晶纤维素复配卡拉胶(1∶1)为稳定剂,添加量为3g/L,制备的中性高钙液体奶对热稳定,口感不会明显增稠,钙含量(以钙元素计)可达2.64g/L。     

Influence of calcium and phosphorus, lactose, and salt-to-moisture ratio on Cheddar cheese quality: pH buffering properties of cheese

The pH buffering capacity of cheese is an important determinant of cheese pH. However, the effects of different constituents of cheese on its pH buffering capacity have not been fully clarified. The objective of this study was to characterize the chemical species and chemical equilibria that are responsible for the pH buffering properties of cheese. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), residual lactose (2.4 vs. 0.78%), and salt-to-moisture ratio (6.4 vs. 4.8%) were manufactured. The pH-titration curves for these cheeses were obtained by titrating cheese:water (1:39 wt/wt) dispersions with 1 N HCl, and backtitrating with 1 N NaOH. To understand the role of different chemical equilibria and the respective chemical species in controlling the pH of cheese, pH buffering was modeled mathematically. The 36 chemical species that were found to be relevant for modeling can be classified as cations (Na⁺, Ca²⁺, Mg²⁺), anions (phosphate, citrate, lactate), protein-bound amino acids with a side-chain pKa in the range of 3 to 9 (glutamate, histidine, serine phosphate, aspartate), metal ion complexes (phosphate, citrate, and lactate complexes of Na⁺, Ca²⁺, and Mg²⁺), and calcium phosphate precipitates. A set of 36 corresponding equations was solved to give the concentrations of all chemical species as a function of pH, allowing the prediction of buffering curves. Changes in the calculated species concentrations allowed the identification of the chemical species and chemical equilibria that dominate the pH buffering properties of cheese in different pH ranges. The model indicates that pH buffering in the pH range from 4.5 to 5.5 is predominantly due to a precipitate of Ca and phosphate, and the protonation equilibrium involving the side chains of protein-bound glutamate. In the literature, the precipitate is often referred to as amorphous colloidal calcium phosphate. A comparison of experimental data and model predictions shows that the buffering properties of the precipitate can be explained, assuming that it consists of hydroxyapatite [Ca₅(OH)(PO₄)₃] or Ca₃(PO₄)₂. The pH buffering in the region from pH 3.5 to 4.5 is due to protonation of side-chain carboxylates of protein-bound glutamate, aspartate, and lactate, in order of decreasing significance. In addition, pH buffering between pH 5 to 8 in the backtitration results from the reprecipitation of calcium and phosphate either as CaHPO₄ or Ca₄H(PO₄)₃.     

Effect of pH on the gel properties and secondary structure of fish myosin

The relationships between gel properties and the secondary structures of silver carp myosin were investigated at pH 5.5–9.0 using dynamic rheological measurement, circular dichroism and scanning electron microscopy. The gel properties of fish myosin were strongly pH and temperature dependent. During heating at 1 °C/min, myosin formed gels in the pH range 5.5–7.5, but not at pH 8.0–9.0. α-Helix was the predominant structure at pH 7.0. The α-helix fraction declined with increasing temperature and the pH away from 7.0, whilst the other secondary structure fractions increased. The α-helix structure of myosin was more susceptive to acid-treatment than alkali-treatment. As pH increased, the gelation rate and gel strength decreased, and the water-holding capacity (WHC) showed an increasing trend followed by a plateau. High β-sheet and β-turn fractions prior to heating could improve G′ at 90 °C, but they depressed the WHC. A compact and uniform gel of fish myosin was obtained at pH 7.0.     

Fortification of reconstituted skim milk powder with different calcium salts: Impact of physicochemical changes on stability to processing

Reconstituted skim milk powder (RSMP) was fortified with 12.5 mM/L calcium (Ca) using soluble [Ca chloride (CChlor), Ca gluconate (CGluc) or Ca hydroxide (CHyd)] or insoluble [Ca carbonate (CCarb), Ca phosphate (CPhos) or Ca citrate (CCit)] salts. CPhos and CCit decreased heat stability moderately at 140 °C, while CCarb had no effect. Soluble salts had a pronounced destabilising effect at 140 °C due to increased ionic Ca levels. After a laboratory‐scale high‐temperature short‐time heating process, CHyd‐fortified RSMP had a lower viscosity than all other samples. CChlor and CGluc increased sedimentation during accelerated physical stability testing, with CHyd causing greater sedimentation than CChlor or CGluc.     

Influence of added calcium chloride on the heat stability of unconcentrated and concentrated bovine milk

The influence of added calcium chloride (1–10 mmol/L) on the heat-induced coagulation of skim bovine milk was examined. Unconcentrated milk displayed a pH-heat coagulation time (HCT) profile with a maximum at pH 6.6 and minimum at pH 7.0. Adding calcium chloride to unconcentrated milk progressively reduced the HCT at the maximum, increased the pH at which the maximum occurred and reduced the HCT at pH > 7.0. For concentrated milk, the shape of the pH-HCT profile, that is, a maximum at pH 6.6, was not altered by added calcium chloride, but HCT was reduced progressively with increasing concentration of calcium chloride. Preheating (90°C for 10 min) shifted the maximum in the pH-HCT profile of unconcentrated milk to a more acidic pH, and addition of 5 mmol/L calcium chloride to preheated milk induced changes in heat stability similar to those noted for unheated milk. Addition of calcium chloride to milk prior to preheating strongly reduced the stability of milk against heat-induced coagulation. These data suggest that calcium has a strong destabilizing effect on the stability of milk systems against heat-induced coagulation.     

Influence of calcium, pH, and moisture on protein matrix structure and functionality in direct-acidified nonfat Mozzarella cheese

Influence of calcium, moisture, and pH on structure and functionality of direct-acid, nonfat Mozzarella cheese was studied. Acetic acid and citric acid were used to acidify milk to pH 5.8 and 5.3 with the aim of producing cheeses with 70 and 66% moisture, and 0.6 and 0.3% calcium levels. Cheeses containing 0.3% calcium were softer and more adhesive than cheeses containing 0.6% calcium, and flowed further when heated. Cheeses with the same calcium content (0.6%), the same moisture content, but set at different pH values (pH 5.3 and 5.8), exhibited no significant differences in melting or firmness. Increasing cheese moisture content from 66 to 70% produced a softer cheese but did not increase meltability. Such differences in functionality corresponded with differences in structure and arrangement of proteins in the cheese protein matrix. Microstructure of cheese with 0.6% calcium had an increase in protein folds and serum pockets compared with the 0.3% calcium cheeses that had a more homogeneous structure. Protein matrix in the low-calcium cheese appeared less dense indicating the proteins were more hydrated. In the 0.6% calcium cheeses, the proteins appeared more aggregated and had larger spaces between protein aggregates. Thus, between pH 5.3 and 5.8, calcium controls cheese functionality, and pH has only an indirect affect related to its influence on the calcium in cheese.