Effect of Buffer Salts on Physical Stability of Lyophilized and Spray-Dried Protein Formulations Containing Bovine Serum Albumin and Trehalose

Calcium-chelating salts (CCS), such as phosphates and citrates, are often added to milk systems to modify physical properties like heat stability. The objective of this study was to investigate the effect of binary CCS mixtures on the properties of casein (CN) micelles including the distribution of Ca between the soluble and CN-bound states. Six binary CCS mixtures were prepared from 4 different types of CCS [i.e., trisodium citrate (TSC), disodium phosphate (DSP), tetrasodium pyrophosphate (TSPP), and sodium hexameta phosphate (SHMP)] by combining 2 CCS at a time in 5 different proportions (8.3:91.7, 29.2:70.8, 50:50, 70.8:29.2, and 91.7:8.3). Different concentrations of these mixtures (0, 0.1, 0.3, 0.5, and 0.7% wt/wt) were added to milk protein concentrate solutions (5% wt/wt) at pH 5.8. The ability of CCS to disperse CN particles and its interaction with Ca were assessed from turbidity measurements, acid-base titration behavior, and the quantity of CN-bound Ca and inorganic phosphate (Pi). Turbidity and the buffering peak at pH ∼5.0 during acid titration decreased with an increasing concentration of CCS. This was due to the chelation of Ca and the dispersion of CN micelles. The presence of TSC in mixtures decreased the amount of CN-bound Ca and Pi; however, the presence of TSPP in mixtures increased CN-bound Ca and Pi. When DSP was present at high proportions in mixtures of CCS, the CN-bound Ca and Pi slightly increased. When SHMP was used in mixtures of CCS, CN-bound Ca and Pi increased with the use of a low proportion of SHMP but decreased when SHMP was used at high proportions in the mixture. Combinations of DSP-TSPP used in the proportions 29.2:70.8, 50:50, and 70.8:29.2 resulted in the gelation of milk protein concentrates when the total CCS concentration was ≥0.3%. These results indicated that the type of CCS present in a mixture modified CN properties by various mechanisms, including chelation of Ca, dispersion of CN micelles, and formation of new types of Ca-CCS complexes. The type of interaction between the newly formed Ca-CCS complexes and the dispersed CN depended on the proportion, concentration, and type of CCS present in the mixtures. This information is useful in understanding how mixtures of CCS affect CN properties.

Background:

Calcium-chelating salts (CCS), such as phosphates and citrates, are often added to milk systems to modify physical properties like heat stability. The objective of this study was to investigate the effect of binary CCS mixtures on the properties of casein (CN) micelles including the distribution of Ca between the soluble and CN-bound states.

Methods:

Six binary CCS mixtures were prepared from 4 different types of CCS [i.e., trisodium citrate (TSC), disodium phosphate (DSP), tetrasodium pyrophosphate (TSPP), and sodium hexameta phosphate (SHMP)] by combining 2 CCS at a time in 5 different proportions (8.3:91.7, 29.2:70.8, 50:50, 70.8:29.2, and 91.7:8.3). Different concentrations of these mixtures (0, 0.1, 0.3, 0.5, and 0.7% wt/wt) were added to milk protein concentrate solutions (5% wt/wt) at pH 5.8. The ability of CCS to disperse CN particles and its interaction with Ca were assessed from turbidity measurements, acid-base titration behavior, and the quantity of CN-bound Ca and inorganic phosphate (Pi).

Results:

Turbidity and the buffering peak at pH ∼5.0 during acid titration decreased with an increasing concentration of CCS. This was due to the chelation of Ca and the dispersion of CN micelles. The presence of TSC in mixtures decreased the amount of CN-bound Ca and Pi; however, the presence of TSPP in mixtures increased CN-bound Ca and Pi. When DSP was present at high proportions in mixtures of CCS, the CN-bound Ca and Pi slightly increased. When SHMP was used in mixtures of CCS, CN-bound Ca and Pi increased with the use of a low proportion of SHMP but decreased when SHMP was used at high proportions in the mixture. Combinations of DSP-TSPP used in the proportions 29.2:70.8, 50:50, and 70.8:29.2 resulted in the gelation of milk protein concentrates when the total CCS concentration was ≥0.3%.

Data Summary:

Turbidity and the buffering peak at pH ∼5.0 decreased with increasing CCS concentration. CN-bound Ca and Pi decreased with TSC, increased with TSPP, slightly increased with high DSP proportions, and showed a biphasic response to SHMP (increased at low proportion, decreased at high proportion). Gelation occurred for DSP-TSPP mixtures at proportions 29.2:70.8, 50:50, and 70.8:29.2 when total CCS ≥0.3% wt/wt.

Conclusions:

These results indicated that the type of CCS present in a mixture modified CN properties by various mechanisms, including chelation of Ca, dispersion of CN micelles, and formation of new types of Ca-CCS complexes. The type of interaction between the newly formed Ca-CCS complexes and the dispersed CN depended on the proportion, concentration, and type of CCS present in the mixtures.

Practical Significance:

This information is useful in understanding how mixtures of CCS affect CN properties.