Effect of trisodium citrate on rheological and physical properties and microstructure of yogurt

The effect of trisodium citrate (TSC) on the rheological and physical properties and microstructure of yogurt was investigated. Reconstituted skim milk was heated at 85° C for 30 min, and various concentrations (5 to 40 mM) of TSC were added to the milk, which was then readjusted to pH 6.50. Milk was inoculated with 2% yogurt culture and incubated at 42° C until pH was 4.6. Acid-base titration was used to determine changes in the state of colloidal calcium phosphate (CCP) in milk. Total and soluble Ca contents of the milk were determined. The storage modulus (G′) and loss tangent (LT) values of yogurts were measured as a function of pH using dynamic oscillatory rheology. Large deformation rheological properties were also measured. Microstructure of yogurt was observed using confocal scanning laser microscopy, and whey separation was also determined. Addition of TSC reduced casein-bound Ca and increased the solubilization of CCP. The G′ value of gels significantly increased with addition of low levels of TSC, and highest G′ values were observed in samples with 10 to 20 mM TSC; higher ( > 20 mM) TSC concentrations resulted in a large decrease in G′ values. The LT of yogurts increased after gelation to attain a maximum at pH ∼5.1, but no maximum was observed in yogurts made with ≥ 25 mM of TSC because CCP was completely dissolved prior to gelation. Partial removal of CCP resulted in an increase in the LT value at pH 5.1. At low TSC levels, the removal of CCP crosslinks may have facilitated greater rearrangement and molecular mobility of the micelle structure, which may have helped to increase G′ and LT values of gels by increasing the formation of crosslinks between strands. At high TSC concentrations the micelles were completely disrupted and CCP crosslinks were dissolved, both of which resulted in very weak yogurt gels with large pores obvious in confocal micrographs. Gelation pH and yield stress significantly decreased with the use of high TSC levels. Lowest whey separation levels were observed in yogurt made with 20 mM TSC, and whey separation greatly increased at > 25 mM TSC. In conclusion, low concentrations of TSC improved several important yogurt characteristics, whereas the use of levels that disrupted casein micelles resulted in poor gel properties. We also conclude that the LT maximum observed in yogurts made from heated milk is due to the presence of CCP because the modification of the CCP content altered this peak and the removal of CCP eliminates this feature in the LT profiles.     

Influence of calcium fortification on sensory, physical and rheological characteristics of fruit yogurt

There has been great demand of calcium fortified dairy products as they can serve as an ideal vehicle for carrying extra calcium to fulfill the nutritional needs but there is need to generate information on the effect of fortification of calcium on the physical properties of these products. In the present study, the calcium enriched mango yogurt was prepared after fortification of pasteurized yogurt mix with 50 mg Ca/100 ml of calcium lactate, this level selected from a preliminary study of sensory evaluation. Fortification of yogurt with calcium lactate at this level significantly (P<0.005) increased the water holding capacity (WHC) by 2.99% on 1st day of storage. WHC of calcium fortified fruit yogurt was higher than control fruit yogurt on 7th and 14th day of storage. Measurements performed on slowly stirred samples (flow curves and final apparent viscosity) showed that calcium-enriched fruit yogurt had stronger structures. Calcium fortified fruit yogurt showed less shear thinning behavior as compared to control. Also, apparent viscosity measurements at constant shear rate showed a significantly (P<0.05) less decrease in initial apparent viscosity in calcium fortified fruit yogurt. However, no statistically significant (P>0.05) difference was observed in tan δ values of control and calcium fortified fruit yogurt indicating similar nature of bonds involved in the gel structure formation of both the yogurt samples. The more firm structure of the calcium fortified fruit yogurt is thus attributed to the higher extent of colloidal calcium phosphate cross-linking between casein micelles due to increased calcium content by fortification. Also flavor, color, and body and texture scores of control and calcium fortified fruit yogurt did not show any significant difference (P>0.05).     

Isolation of caseins from whey proteins by microfiltration modifying the mineral balance in skim milk

The objective of this work was to study the effect of different salts and salt concentration on the isolation of casein micelles from bovine raw skim milk by tangential flow microfiltration. Tangential flow microfiltration (0.22 μm) was conducted in a continuous process adding a modified buffer to maintain a constant initial sample volume. This buffer contained calcium chloride (CaCl₂), sodium phosphate (Na₂HPO₄), or potassium citrate (K₃C₆H₅O₇) in concentrations ranging from 0 to 100 mM. The concentrations of caseins and whey proteins retained were determined by sodium dodecyl sulfate-PAGE and analyzed using the Scion Image software (Scion Corporation, Frederick, MD). A complete isolation of caseins from whey proteins was achieved using sodium phosphate in the range of 10 to 50 mM and 20 times the initial volume of buffer added. No whey proteins were detected at 50 mM but this was at the expense of low caseins being retained. When lower sodium phosphate concentrations were used, the amount of caseins retained was higher but a small amount of whey proteins were still detected by sodium dodecyl sulfate-PAGE. Among the salts tested, calcium chloride at 50 mM and all volumes of buffer showed the higher retention of casein proteins. The highest casein:whey protein ratio was found at 30 mM CaCl₂, but no complete casein micelle isolation was achieved. Potassium citrate was the most ineffective salt because a rapid loss of caseins and whey proteins was observed at all concentrations and with low quantities of buffer added during the filtration process. Our results show the potential of altering the mineral balance in milk for isolation of casein micelles from whey proteins in a continuous tangential flow microfiltration system.     

Changes in physical properties of bovine milk from the colostrum period to early lactation

The aim of this study was to analyze individual cows’ samples from the colostrum, postcolostrum, and early lactation periods to investigate how milk composition, physical properties, stability, and suitability for processing change throughout this period. Attention was paid to the first week postpartum in which the composition of bovine mammary secretion can change markedly. Properties including pH, titratable acidity, ethanol stability (ES), rennet clotting time, and casein micelle size were analyzed, together with some compositional factors such as fat, total protein, lactose, total and ionic calcium, magnesium, citrate, phosphorus, sodium, and potassium. Total Ca (36.2 mM) and free ionic Ca (2.58 mM), Mg (5.9 mM), P (32.2 mM), and Na (24.1 mM) appeared to be high on d 5 postpartum, having decreased substantially over the first 5 d; they gradually decreased thereafter. The average pH on d 5 was only 6.49, compared with 6.64 at 1 mo postpartum. Stability measurements showed that the average ES on d 5 was 70% and the rennet clotting time was 12.2 min, which were significantly lower than values at later stages. A number of milk properties including ES, pH, protein content, and Ca²⁺ concentration could be useful for identifying the point of transition from colostrum to the early lactation period. Knowing the composition and physical properties of colostrum and postcolostrum secretions will help establish when such milk is suitable for processing and determine the best use for that milk.     

Properties of milk protein gels formed by phosphates

We investigated the properties of gels that were formed by adding emulsifying salts, such as tetrasodium pyrophosphate (TSPP), to reconstituted milk protein concentrate solution. The pH of a 51 g/L milk protein concentrate solution was adjusted to 5.8 after adding TSPP. Milk protein concentrate solutions were placed in glass jars and allowed to stand at 25°C for 24 h. Gels with the highest breaking force were formed when TSPP was added at a concentration of 6.7 mM, whereas no gel was formed when TSPP was added at concentrations of ≤2.9 or ≥10.5 mM. Several other phosphate-based emulsifying salts were tested but for these emulsifying salts, gelation only occurred after several days or at greater gelation temperatures. No gelation was observed for trisodium citrate. Gelation induced by TSPP was dependent on pH, and the breaking force of gel was greatest at pH 6.0. Furthermore, when the concentration of milk protein concentrate in solution was increased to 103 g/L, the breaking force of the gel increased, and a clearly defined network between caseins could be observed by using confocal scanning laser microscopy. These results suggest that TSPP-induced gelation occurs when the added TSPP acts with calcium as a cross-linking agent between dispersed caseins and when the balance between (a reduced) electrostatic repulsion and (enhanced) attractive (hydrophobic) interactions becomes suitable for aggregation and eventual gelation of casein molecules.     

Enrichment of stirred yogurt with soluble dietary fiber from Pachyrhizus erosus L. Urban: Effect on syneresis, microstructure and rheological properties

Yam soluble fiber (YSF) extracted from Pachyrhizus erosus was added (1 g per 100 mL) to a stirred yogurt (SY_YSF). Its syneresis and microstructure properties were evaluated and compared to those of a stirred yogurt (SY_C) without added YSF. The SY_C yogurt exhibited a more compact casein micelle aggregates network than that of the SY_YSF yogurt which was more open, relaxed and covered with fibrous structures attributed to the YSF components. The rheological analysis showed that the SY_YSF yogurt had lower storage modulus (G′) and loss modulus (G″) values in the linear viscoelastic region than the SY_C yogurt, but its flow behavior was characterized by a lower flow index (n), higher consistency index (k), and higher yield stress (τ₀) than the SY_C yogurt. Incorporation of the YSF reduced significantly the syneresis and produced a more acceptable mouthfeel in the SY_YSF yogurt in comparison to the SY_C yogurt, indicating the viability of the process to obtain a commercial product.     

Determination of calcium-binding constants of caseins,phosphoserine, citrate and pyrophosphate: A modelling approach using free calcium measurement

Dairy products contain large amount of calcium which is bound to caseins and different chelating agents like citrate and polyphosphates. The present study aimed to determine the calcium-binding capacities of phosphoserine (SerP), caseinophosphopeptide (CPP), β-casein, caseinate, citrate and pyrophosphate in the same conditions of temperature, pH and ionic strength. The free calcium (Ca²⁺) was measured using a calcium ion-selective electrode and plotted as a function of total calcium concentration. The association constants and the number of calcium-binding sites were determined by fitting the experimental data to a theoretical model. The phosphate groups of caseins were the main binding sites with evidence for participation of carboxylate groups. The intrinsic association constants determined by the best fit of the data were in the order: pyrophosphate (557 × 10³ M⁻¹) > citrate (20 × 10³ M⁻¹) > β-casein (5 × 10³ M⁻¹) > caseinate, CPP and SerP (∼10³ M⁻¹). These findings may be of interest for the development of calcium-enriched products to overcome calcium deficiency in specific populations.     

Use of weak cation exchange resin Lewatit S 8528 as alternative to strong ion exchange resins for calcium salt removal

Softening of beet sugar juices using the weak cation exchange resin Lewatit® S 8528, both in hydrogen and sodium form has been studied. After analyzing the influence of the main operating parameters on decalcification efficiencies through a Taguchi approach, experimental runs were carried out at 40 BV/h and 80 °C to determine operation time, breakthrough point and resin capacity. The performance of the Lewatit S 8528 resin was compared with that of the industrially used Amberlite SR1L Na, a strong cation exchange resin. The experimental results showed that the use of Lewatit S 8528 in the acidic form is a feasible alternative to the classical strong cation exchange resins, providing calcium removal efficiencies higher than 80%. The mean calcium concentration in the softened thin juice was lower than 18 mg CaO/L. The breakthrough point corresponding to weak resin Lewatit S 8528 in H⁺ form, 700 bed volume, was considerable higher than that obtained for the strong resin Amberlite (300 BV). No considerably increase in the concentration of reducing sugars in the softened juice was observed. The consumption of chemical reagents in the regeneration stage was 3.2 ± 0.5 kg H₂SO₄/m³, considerably lower than the consumption of sodium chloride, 44 ± 1 kg NaCl/m³ when the strong Amberlite resin was used.     

Industrial yogurt manufacture: monitoring of fermentation process and improvement of final product quality

Lactic acid fermentation during the production of skim milk and whole fat set-style yogurt was continuously monitored by measuring pH. The modified Gompertz model was successfully applied to describe the pH decline and viscosity development during the fermentation process. The viscosity and incubation time data were also fitted to linear models against ln(pH). The investigation of the yogurt quality improvement practices included 2 different heat treatments (80°C for 30 min and 95°C for 10 min), 3 milk protein fortifying agents (skim milk powder, whey powder, and milk protein concentrate) added at 2.0%, and 4 hydrocolloids (κ-carrageenan, xanthan, guar gum, and pectin) added at 0.01% to whole fat and skim yogurts. Heat treatment significantly affected viscosity and acetaldehyde development without influencing incubation time and acidity. The addition of whey powder shortened the incubation time but had a detrimental effect on consistency, firmness, and overall acceptance of yogurts. On the other hand, addition of skim milk powder improved the textural quality and decreased the vulnerability of yogurts to syneresis. Anionic stabilizers (κ-carrageenan and pectin) had a poor effect on the texture and palatability of yogurts. However, neutral gums (xanthan and guar gum) improved texture and prevented the wheying-off defect. Skim milk yogurts exhibited longer incubation times and higher viscosities, whereas they were rated higher during sensory evaluation than whole fat yogurts.     

Thermal stability of soy protein isolate and hydrolysate ingredients

The objective of this study was to characterize the effects of pH, protein concentration and calcium supplementation on thermal stability, at 140 °C, of soy protein isolate (SPI) and soy protein hydrolysate (SPH) ingredients. Increasing pH between 6.4 and 7.5 led to significantly (p < 0.05) higher mean heat coagulation times (HCTs) at 140 °C, for all soy protein ingredients at 1.8, and 3.6% (w/v) protein. Increasing protein concentration from 1.8 to 7.2% (w/v) led to shorter HCTs for protein dispersions. Calcium supplementation up to 850 mg/L, except in the case of supplementation of SPI 1 with calcium citrate (CaCit), decreased HCT for soy protein ingredient dispersions, at pH 6.4 – 7.5. No significant differences (p < 0.05) were found in mean HCT for dispersions supplemented with calcium chloride (CaCl₂) and those supplemented with CaCit at 450, 650 and 850 mg/L Ca²⁺, in the pH range 6.4–7.5.     

Survival of Lactobacillus acidophilus and Bifidobacterium animalis ssp. lactis in stirred fruit yogurts

The effect of commercial fruit preparations (mango, mixed berry, passion fruit and strawberry) on the viability of probiotic bacteria, Lactobacillus acidophilus LAFTI^® L10 and Bifidobacterium animalis ssp. lactis LAFTI^® B94 in stirred yogurts during storage (35 days) at refrigerated temperature (4 °C) was evaluated. The results showed that addition of either 5 or 10 g/100 g fruit preparations had no significant (p>0.05) effect on the viability of the two probiotic strains except on L. acidophilus LAFTI L10 yogurt with 10 g/100 g passion fruit or mixed berry. After the addition of fruit preparation, 96% of the yogurts incorporated with fruit preparation did not exhibit a greater loss in the viability of probiotic bacteria compared to plain yogurt during the storage period. A correlation between the post-storage pH in yogurts and the survival of probiotic bacteria was observed. All the yogurts, however, contained the recommended levels of (10⁶-10⁷ cfu/g) probiotic bacteria at the end of 35-day shelf life.     

Cell wall-modifying enzymes and firmness loss in ripening ‘Golden Reinders’ apples: A comparison between calcium dips and ULO storage

Calcium treatment and storage under ultra-low oxygen (ULO) conditions are common post-harvest practices aimed at delaying ripening-related softening of apple (Malus × domestica Borkh.) fruit, but the biochemical mechanisms underlying these effects have not been determined conclusively to date. In this study, commercially mature ‘Golden Reinders’ apples were dipped in 2% calcium chloride prior to storage at 1 °C and 92% RH under either regular air or ultra-low oxygen (ULO; 1kPa O₂:2kPa CO₂) for 19 or 31 weeks, and kept thereafter at 20 °C for 0, 7 or 14 days in order to simulate the usual marketing time. Cell wall composition and cell wall-modifying enzyme activities were determined in relation to fruit firmness. ULO-storage and calcium dips were effective for firmness preservation, seemingly due to decreased pectin solubilisation. β-Galactosidase, α-l-arabinofuranosidase and pectate lyase activities were correlated positively with firmness loss of ‘Golden Reinders’ fruit after 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.     

Determination of antioxidant activity of bioactive peptide fractions obtained from yogurt

In this study, physicochemical and microbiological properties of traditional and commercial yogurt samples were determined during 4 wk of storage. Proteolytic activity, which occurs during the storage period of yogurt samples, was also determined. Peptide fractions obtained from yogurts were investigated and the effect of proteolysis on peptide release during storage was determined. The antioxidant activities of peptides released from yogurt water-soluble extracts (WSE) and from HPLC fractions were determined by 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) methods. The antioxidant activity of WSE from traditional yogurt was greater than that of WSE from commercial yogurts. In analysis by the ABTS method, mean values increased from 7.697 to 8.739 mM Trolox/g in commercial yogurts, and from 10.115 to 13.182 mM Trolox/g in traditional yogurts during storage. Antioxidant activities of peptides released from HPLC fractions of selected yogurt samples increased 10 to 200 times. In all yogurt samples, the greatest antioxidant activity was shown in the F2 fraction. After further fractionation of yogurt samples, the fractions coded as F2.2, F2.3, F4.3, and F4.4 had the highest antioxidant activity values. Total antioxidant activity of yogurts was low but after purification of peptides by fractionation in HPLC, peptide fractions with high antioxidant activity were obtained.     

Effect of calcium chloride addition on ice cream structure and quality

The influence of calcium fortification by the addition of calcium chloride on quality parameters of ice cream based on physical properties was investigated, as was the effect of κ-carrageenan at modifying the effects of this calcium fortification. Four ice cream mixes of conventional composition, with added κ-carrageenan (0 or 0.025%) and added calcium chloride (0 or 4.4 g L⁻¹ = 40 mM of added Ca²⁺), were prepared. Modulated temperature-differential scanning calorimetry was used to investigate the effect of calcium chloride on the nucleation temperature, enthalpy of melting, and freezing point depression. The protein composition of 15.4% (wt/wt) reconstituted skim milk powder solutions with or without 4.4 g L⁻¹ added CaCl₂ and in the supernatant after ultracentrifugation was determined. Fat particle size distributions in ice cream were characterized by light scattering. Ice crystal sizes before and after temperature cycling were determined by cold-stage light microscopy. The results demonstrated that the addition of calcium chloride led to a substantial increase in ice crystal sizes and in fat partial coalescence, which were exacerbated by the addition of κ-carrageenan. These results can be explained by the interaction between Ca²⁺ ions and casein micelles, rather than any effects on freezing point depression. The calcium ions led to a more compact micelle, less serum β-casein, and high fat destabilization, all of which would be expected to reduce macromolecular structure and volume occupancy in the unfrozen phase, which led to increased rates of ice recrystallization.     

Evaluation of physical properties during storage of set and stirred yogurts made from ultra-high pressure homogenization-treated milk

The effects of ultra-high pressure homogenization (UHPH) on cow's milk were investigated and its suitability for yogurt manufacturing was compared with the conventional process currently applied in the yogurt industry. Yogurts were prepared from UHPH-treated milks at 200 and 300 MPa at 40 °C, and yogurts prepared from heat-treated milk at 90 °C for 90 s, homogenized at 15 MPa and enriched with 3% of skim milk powder were used as control samples. This study included determination of titrable acidity, water-holding capacity (WHC), and textural and rheological evaluation of gels in both set-type and stirred yogurts. In order to follow the evolution of yogurts during storage at refrigeration temperature (4–6 °C), all analyses were carried out weekly (1, 7, 14, 21 and 28 days). Results showed that yogurts from UHPH-treated milk presented higher WHC and firmness values compared with the conventional yogurts. However, the disruption of the network from UHPH-treated milk into stirred gels resulted in yogurts with higher consistency, less syneresis but coarser structure than the conventional ones.     

Evidence for fibril-like structure in bovine casein micelles

The addition of Congo red (CR) dye to diluted raw skim milk resulted in a red shift indicative of the presence of fibril-like structures. Thioflavin T (ThT) is another dye that very specifically binds to protein fibrils, and when added to undiluted raw skim milk, the classic 485 nm fluorescence peak of a ThT-fibril complex was observed. Repeating these experiments with various raw milk components showed that the CR red shift and ThT fluorescence peak were due to the presence of casein micelles, and to a lesser extent, sodium caseinate. Fluorescent peaks were also observed when ThT was added to solutions of purified α_S- and κ-casein, but not β-casein, in 0.5 M HEPES buffer (pH = 6.8). The addition of 25 mM Ca²⁺ had no effect on β-casein fluorescence, and significantly reduced the κcasein peak. However, adding 25 mM Ca²⁺ to α_S-casein produced a turbid solution and a 6-fold increase in fluorescence, indicating that the aggregates formed contain fibril-like structure. Casein micelle images obtained by transmission electron microscopy showed the presence of short (7 to 10 nm) fibers cross-linked by dense aggregate junction zones. The observed fibers closely resemble protofibrils, intermediate structures that are observed during the formation of amyloid fibrils.     

Individual and sequential effects of stirring,smoothing, and cooling on the rheological properties of nonfat yogurts stirred with a technical scale unit

Few studies have considered the impact of unit operations during stirred yogurt manufacture because their operational sequence is difficult to replicate at the laboratory scale. The aim of this study was to investigate the individual and sequential effects of stirring in the yogurt vat, smoothing, and cooling on the rheological properties of yogurts, using a technical scale unit simulating some industrial conditions. The yogurts were prepared from a milk mixture that was standardized to contain 14% total solids, 0% fat, and 4% protein, and then homogenized, heated (94.5°C, 5 min), and inoculated at 41°C with the same thermophilic lactic starter. The operating parameters under investigation were 2 stirring durations in the yogurt vat (5 or 10 min), 2 cooling systems (plate or tubular heat exchanger), and 2 smoothing temperatures (38°C for smoothing before cooling; 20°C for smoothing after cooling). Sampling valves were installed at critical points on the technical scale unit so that the effect of each operation on the properties of stirred yogurt could be quantified individually. Syneresis, apparent viscosity, firmness, and consistency were analyzed after 1 d of storage at 4°C. In general, as the yogurts moved through the technical scale unit, the properties of the yogurts (evaluated after 1 d) changed: viscosity increased but syneresis, firmness, and consistency decreased. The individual effects of the operations showed that smoothing and cooling, compared with stirring duration, made the greatest contribution in terms of modifying yogurt properties. The stirring parameters (5 or 10 min) had similar effects on the yogurts. The use of a plate heat exchanger promoted a decrease in syneresis, whereas a tubular heat exchanger had a greater effect in terms of increasing firmness and consistency. The type of cooling system had no effect on stirred yogurt viscosity. Smoothing at 38°C had a greater effect on the increase in firmness, whereas smoothing at 20°C contributed more to a decrease in syneresis and increases in viscosity and consistency. This study confirms that each unit operation has a defined effect on the rheological properties of a nonfat stirred yogurt, which also depends on the operation sequence.