Tenderness, juiciness, colour and flavour are the most important meat quality attributes affecting the consumer acceptance. Maintaining the consistency of meat products by avoiding variable quality has become a major concern and great challenge to the meat industry. This in turn will also benefit meat end-users in the marketplace by having more tender meat. The present study was designed to evaluate the overall effects of pre-rigor infusion with kiwifruit juice, which contains the plant protease, actinidin, on lamb quality. A total of 18 lambs (12 months old) were divided into three treatment groups (6 lambs per each treatment). After exsanguination, lamb carcasses were infused (10% body weight) with fresh kiwifruit juice (Ac), water (W) and compared with a noninfusion treatment which acted as a control (C). Samples from different muscle/cuts (longissimus dorsi (LD) vs leg chops) at different post-mortem times (1 day post-mortem vs. 3 wks vacuum packaged storage at 2°C) and display time (0 to 6 days after the post-mortem storage) were analysed to monitor the changes on meat physical properties (e.g., tenderness, temperature, drip and cooking loss, colour), biochemical changes (pH, proteins and lipids) and volatile flavour compounds after the infusion treatments. The most tender meat (lowest shear force values) (P < 0.001) detected in the Ac carcasses post-mortem compared with C and W carcasses demonstrated that kiwifruit juice was a very powerful meat tenderizer, and could contribute to the meat tenderization process efficiently and effectively. Compared with C and W carcasses, the enhanced proteolytic activity (P = 0.002) resulting from the actinidin in kiwifruit juice in Ac carcasses caused degradation of the myofibrillar proteins and the appearance of new peptides during postmortem ageing. A slight positive effect in a*-value (redness) and decreased lipid oxidation, found in leg chops, was thought to be caused by the natural antioxidants in kiwifruit juice. Kiwifruit juice infused into the meat did not alter (P > 0.05) the volatile flavour compound profile indicating that the meat from Ac treated carcasses maintained its natural lamb flavour. No treatment differences were found for the temperature decline (P > 0.05) between the infused treatments and C. The higher rate of pH decline (P < 0.05) found in W carcasses might have contributed to the higher drip and cooking loss. The unbound water in meat might contribute to the higher L*-values (lightness) found in W carcasses. In summary, the proteolytic tenderizing infusion treatment using kiwifruit juice is a feasible approach for the commercial meat industry to increase profits, and also could satisfy the eating quality standards required by the consumers. In addition, tenderizing meat by using kiwifruit juice could also provide the kiwifruit processors an additional option for use of their product to gain a more profitable return.

The present study was aimed at extending our knowledge of the antioxidative properties of the milk protein products, whey protein isolate (WPI) and sodium caseinate (NaCas), in oil-in-water (O/W) emulsions rich in polyunsaturated fatty acids (PUFAs). In particular, the objective was to contribute to our understanding of the compositional and processing factors that influence the oxidative stability of protein-stabilised O/W emulsions. Linoleic acid (approximately 60 %) was used as the lipid for the oil phase (10.6 %). The emulsion samples were usually incubated at 50 °C to accelerate lipid oxidation. Lipid oxidation indicators were lipid hydroperoxides and headspace hexanal, determined by solid phase microextraction (SPME) combined with gas chromatography (GC). WPI- or NaCas-stabilised emulsions were prepared using a wide range of protein concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 7.0 or 10.0 %) at two droplet sizes (d32 = 0.31 and 0.65 µm). In general, higher lipid oxidation levels were found for the larger droplet size. Increasing protein concentration led to a decrease in the lipid oxidation rate. The greatest decrease in lipid hydroperoxide levels (values after 4 h) occurred at up to 4.0 % protein concentration. The greatest decrease in hexanal levels (values after 24 h) occurred at up to 4.0 % protein concentration in WPI emulsions (0.31 µm). The hexanal levels were more independent of the protein concentration in the other emulsion types. The hexanal level decreased at protein concentrations > 4.0 % in NaCas emulsions (0.31 and 0.65 µm) and at protein concentrations > 7.0 % in WPI emulsions (0.65 µm). The difference between lipid hydroperoxide generation in emulsions with small and large droplet sizes decreased with increasing protein concentration. This effect was more pronounced in NaCas emulsions. In general, NaCas was a better inhibitor of lipid oxidation than WPI, but WPI appeared to be the better antioxidant at some droplet size/protein concentration combinations. The protein in the continuous phase, i.e. the unadsorbed protein, played an important role in lipid oxidation. In principal, the lipid hydroperoxide and hexanal levels showed the same development over the continuous phase protein concentration as over the protein concentration in WPI and NaCas emulsions (d32 = 0.31 µm). A low NaCas level in the continuous phase already led to a relatively low hexanal level, whereas a higher WPI level was required. When NaCas solution was added to a WPI emulsion or WPI solution was added to a NaCas emulsion, a synergistic antioxidative effect was observed. The high molecular weight fractions (molecular weight = 12000-14000) of WPI and NaCas contained pro-oxidative metal ions that contributed to lipid oxidation in the emulsions. An enrichment of NaCas emulsions with the low molecular weight fraction of NaCas (with a molecular weight = 12000-14000) notably inhibited lipid oxidation. An enrichment of WPI emulsions with the low molecular weight fraction of WPI (with a molecular weight = 12000-14000) also seemed to inhibit lipid oxidation, but the effect was not significant. The protein solutions were enriched with these fractions before emulsion preparation. Pure WPI solution or mixed WPI/NaCas (1:1, weight/weight) solution with 1.12 or 2.24 % protein concentration was heated at 84 °C for up to 40 min, cooled and then used to prepare emulsions. Lipid oxidation was generally not affected by the heat treatment or the degree of whey protein denaturation. However, at the lower WPI concentration, more hexanal was produced for the longer heating times (20, 30 and 40 min) and this appeared to be connected with the physical instability of the emulsions. Greater oxidative stability was found at the higher protein concentration and when the proteins were mixed, pointing to a possible synergistic antioxidative effect of WPI and NaCas. The addition of the free radical source 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH) greatly increased the oxygen uptake and the generation of lipid hydroperoxides in the emulsions. The oxidative stability increased with increasing protein concentration (1.0, 4.0 and 7.0 %). NaCas had a greater antioxidative effect than WPI. The inhibition of oxygen uptake appeared to be largely influenced by the free-radical-scavenging activity of the system, determined by the protein type and the protein concentration, as the radicals were produced linearly over time and oxygen was consumed linearly over time. It can therefore be concluded that free-radical-scavenging activity represents a major antioxidative mechanism of the milk proteins. Oxygen was consumed much faster in emulsions than in protein solutions when the same level of AAPH was incorporated. In a WPI (1.0 % protein) emulsion, much lower levels of protein hydroperoxides than of lipid hydroperoxides developed. This pointed to a much greater reactivity of linoleic acid than of the milk proteins with oxygen. In contrast, the exposure of WPI to oxidising linoleic acid in an emulsion (1.0 % protein) or to AAPH in aqueous solution led to oxidative damage of the whey proteins, indicated by the loss of amino acids. The loss of specific amino acids was different for proteins in the continuous phase or cream phase of an emulsion or in WPI solution. The present study confirms the antioxidative potential of WPI and NaCas and gives new insights into their functionality as oxidative stabilisers in O/W emulsions.

Chronic graft dysfunction (CGD) is an important post-transplant complication. CGD can be considered as an impaired repair process, which ultimately leads to the loss of graft function.To study non-immunological factors contributing to the development of CGD in kidney grafts we used in vitro and in vivo models, and clinical studies. We studied the actions of hyperlipidemia in vitro. LDL induced increased expression of TGF-β1 and TGF-β receptors type I and type II. Smad2 phosphorylation could be induced by conditioned medium from mesangial cells incubated with LDL. The effects of Fluvastatin and AT1 receptor blocker Candesartan cilexetil on aortic graft arteriosclerosis in the rat were evaluated. Fluvastatin neither alone nor in combination with Cyclosporine A affected allograft remodelling, but reduced neointima formation in isografts. Candesartan cilexetil treatment reduced graft arteriosclerosis. The effect is explained by the reduction of TGF-β1 expression. We investigated the effects of Carvedilol in patients with CGD. Carvedilol failed to alter the CGD progression despite the efficient control of blood pressure, and a beneficial effect on lipid pattern and oxidation. Close control of CyA blood levels is recommended due to interaction between CyA and Carvedilol. Measurement of Ab-oxLDL in kidney graft recipients demonstrated that these patients had lower Ab-oxLDL levels as compared with the control group. Decreased Ab-oxLDL levels were associated with graft loss due to acute rejection and with ischemic heart disease. In this thesis we have addressed several important complex issues, which are interconnected: (1) development of chronic graft dysfunction (2) lipoproteins and their role in inducing pathological conditions like atherosclerosis and graft damage, (3) oxidation, (4) TGF-β and its' role in different pathological conditions, including renal and vascular damage.

The present study was aimed at extending our knowledge of the antioxidative properties of the milk protein products, whey protein isolate (WPI) and sodium caseinate (NaCas), in oil-in-water (O/W) emulsions rich in polyunsaturated fatty acids (PUFAs). In particular, the objective was to contribute to our understanding of the compositional and processing factors that influence the oxidative stability of protein-stabilised O/W emulsions. Linoleic acid (approximately 60 %) was used as the lipid for the oil phase (10.6 %). The emulsion samples were usually incubated at 50 °C to accelerate lipid oxidation. Lipid oxidation indicators were lipid hydroperoxides and headspace hexanal, determined by solid phase microextraction (SPME) combined with gas chromatography (GC). WPI- or NaCas-stabilised emulsions were prepared using a wide range of protein concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 7.0 or 10.0 %) at two droplet sizes (d32 = 0.31 and 0.65 µm). In general, higher lipid oxidation levels were found for the larger droplet size. Increasing protein concentration led to a decrease in the lipid oxidation rate. The greatest decrease in lipid hydroperoxide levels (values after 4 h) occurred at up to 4.0 % protein concentration. The greatest decrease in hexanal levels (values after 24 h) occurred at up to 4.0 % protein concentration in WPI emulsions (0.31 µm). The hexanal levels were more independent of the protein concentration in the other emulsion types. The hexanal level decreased at protein concentrations > 4.0 % in NaCas emulsions (0.31 and 0.65 µm) and at protein concentrations > 7.0 % in WPI emulsions (0.65 µm). The difference between lipid hydroperoxide generation in emulsions with small and large droplet sizes decreased with increasing protein concentration. This effect was more pronounced in NaCas emulsions. In general, NaCas was a better inhibitor of lipid oxidation than WPI, but WPI appeared to be the better antioxidant at some droplet size/protein concentration combinations. The protein in the continuous phase, i.e. the unadsorbed protein, played an important role in lipid oxidation. In principal, the lipid hydroperoxide and hexanal levels showed the same development over the continuous phase protein concentration as over the protein concentration in WPI and NaCas emulsions (d32 = 0.31 µm). A low NaCas level in the continuous phase already led to a relatively low hexanal level, whereas a higher WPI level was required. When NaCas solution was added to a WPI emulsion or WPI solution was added to a NaCas emulsion, a synergistic antioxidative effect was observed. The high molecular weight fractions (molecular weight = 12000-14000) of WPI and NaCas contained pro-oxidative metal ions that contributed to lipid oxidation in the emulsions. An enrichment of NaCas emulsions with the low molecular weight fraction of NaCas (with a molecular weight = 12000-14000) notably inhibited lipid oxidation. An enrichment of WPI emulsions with the low molecular weight fraction of WPI (with a molecular weight = 12000-14000) also seemed to inhibit lipid oxidation, but the effect was not significant. The protein solutions were enriched with these fractions before emulsion preparation. Pure WPI solution or mixed WPI/NaCas (1:1, weight/weight) solution with 1.12 or 2.24 % protein concentration was heated at 84 °C for up to 40 min, cooled and then used to prepare emulsions. Lipid oxidation was generally not affected by the heat treatment or the degree of whey protein denaturation. However, at the lower WPI concentration, more hexanal was produced for the longer heating times (20, 30 and 40 min) and this appeared to be connected with the physical instability of the emulsions. Greater oxidative stability was found at the higher protein concentration and when the proteins were mixed, pointing to a possible synergistic antioxidative effect of WPI and NaCas. The addition of the free radical source 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH) greatly increased the oxygen uptake and the generation of lipid hydroperoxides in the emulsions. The oxidative stability increased with increasing protein concentration (1.0, 4.0 and 7.0 %). NaCas had a greater antioxidative effect than WPI. The inhibition of oxygen uptake appeared to be largely influenced by the free-radical-scavenging activity of the system, determined by the protein type and the protein concentration, as the radicals were produced linearly over time and oxygen was consumed linearly over time. It can therefore be concluded that free-radical-scavenging activity represents a major antioxidative mechanism of the milk proteins. Oxygen was consumed much faster in emulsions than in protein solutions when the same level of AAPH was incorporated. In a WPI (1.0 % protein) emulsion, much lower levels of protein hydroperoxides than of lipid hydroperoxides developed. This pointed to a much greater reactivity of linoleic acid than of the milk proteins with oxygen. In contrast, the exposure of WPI to oxidising linoleic acid in an emulsion (1.0 % protein) or to AAPH in aqueous solution led to oxidative damage of the whey proteins, indicated by the loss of amino acids. The loss of specific amino acids was different for proteins in the continuous phase or cream phase of an emulsion or in WPI solution. The present study confirms the antioxidative potential of WPI and NaCas and gives new insights into their functionality as oxidative stabilisers in O/W emulsions.

The present study was aimed at extending our knowledge of the antioxidative properties of the milk protein products, whey protein isolate (WPI) and sodium caseinate (NaCas), in oil-in-water (O/W) emulsions rich in polyunsaturated fatty acids (PUFAs). In particular, the objective was to contribute to our understanding of the compositional and processing factors that influence the oxidative stability of protein-stabilised O/W emulsions. Linoleic acid (approximately 60 %) was used as the lipid for the oil phase (10.6 %). The emulsion samples were usually incubated at 50 °C to accelerate lipid oxidation. Lipid oxidation indicators were lipid hydroperoxides and headspace hexanal, determined by solid phase microextraction (SPME) combined with gas chromatography (GC). WPI- or NaCas-stabilised emulsions were prepared using a wide range of protein concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 7.0 or 10.0 %) at two droplet sizes (d32 = 0.31 and 0.65 µm). In general, higher lipid oxidation levels were found for the larger droplet size. Increasing protein concentration led to a decrease in the lipid oxidation rate. The greatest decrease in lipid hydroperoxide levels (values after 4 h) occurred at up to 4.0 % protein concentration. The greatest decrease in hexanal levels (values after 24 h) occurred at up to 4.0 % protein concentration in WPI emulsions (0.31 µm). The hexanal levels were more independent of the protein concentration in the other emulsion types. The hexanal level decreased at protein concentrations > 4.0 % in NaCas emulsions (0.31 and 0.65 µm) and at protein concentrations > 7.0 % in WPI emulsions (0.65 µm). The difference between lipid hydroperoxide generation in emulsions with small and large droplet sizes decreased with increasing protein concentration. This effect was more pronounced in NaCas emulsions. In general, NaCas was a better inhibitor of lipid oxidation than WPI, but WPI appeared to be the better antioxidant at some droplet size/protein concentration combinations. The protein in the continuous phase, i.e. the unadsorbed protein, played an important role in lipid oxidation. In principal, the lipid hydroperoxide and hexanal levels showed the same development over the continuous phase protein concentration as over the protein concentration in WPI and NaCas emulsions (d32 = 0.31 µm). A low NaCas level in the continuous phase already led to a relatively low hexanal level, whereas a higher WPI level was required. When NaCas solution was added to a WPI emulsion or WPI solution was added to a NaCas emulsion, a synergistic antioxidative effect was observed. The high molecular weight fractions (molecular weight = 12000-14000) of WPI and NaCas contained pro-oxidative metal ions that contributed to lipid oxidation in the emulsions. An enrichment of NaCas emulsions with the low molecular weight fraction of NaCas (with a molecular weight = 12000-14000) notably inhibited lipid oxidation. An enrichment of WPI emulsions with the low molecular weight fraction of WPI (with a molecular weight = 12000-14000) also seemed to inhibit lipid oxidation, but the effect was not significant. The protein solutions were enriched with these fractions before emulsion preparation. Pure WPI solution or mixed WPI/NaCas (1:1, weight/weight) solution with 1.12 or 2.24 % protein concentration was heated at 84 °C for up to 40 min, cooled and then used to prepare emulsions. Lipid oxidation was generally not affected by the heat treatment or the degree of whey protein denaturation. However, at the lower WPI concentration, more hexanal was produced for the longer heating times (20, 30 and 40 min) and this appeared to be connected with the physical instability of the emulsions. Greater oxidative stability was found at the higher protein concentration and when the proteins were mixed, pointing to a possible synergistic antioxidative effect of WPI and NaCas. The addition of the free radical source 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH) greatly increased the oxygen uptake and the generation of lipid hydroperoxides in the emulsions. The oxidative stability increased with increasing protein concentration (1.0, 4.0 and 7.0 %). NaCas had a greater antioxidative effect than WPI. The inhibition of oxygen uptake appeared to be largely influenced by the free-radical-scavenging activity of the system, determined by the protein type and the protein concentration, as the radicals were produced linearly over time and oxygen was consumed linearly over time. It can therefore be concluded that free-radical-scavenging activity represents a major antioxidative mechanism of the milk proteins. Oxygen was consumed much faster in emulsions than in protein solutions when the same level of AAPH was incorporated. In a WPI (1.0 % protein) emulsion, much lower levels of protein hydroperoxides than of lipid hydroperoxides developed. This pointed to a much greater reactivity of linoleic acid than of the milk proteins with oxygen. In contrast, the exposure of WPI to oxidising linoleic acid in an emulsion (1.0 % protein) or to AAPH in aqueous solution led to oxidative damage of the whey proteins, indicated by the loss of amino acids. The loss of specific amino acids was different for proteins in the continuous phase or cream phase of an emulsion or in WPI solution. The present study confirms the antioxidative potential of WPI and NaCas and gives new insights into their functionality as oxidative stabilisers in O/W emulsions.

The present study was aimed at extending our knowledge of the antioxidative properties of the milk protein products, whey protein isolate (WPI) and sodium caseinate (NaCas), in oil-in-water (O/W) emulsions rich in polyunsaturated fatty acids (PUFAs). In particular, the objective was to contribute to our understanding of the compositional and processing factors that influence the oxidative stability of protein-stabilised O/W emulsions. Linoleic acid (approximately 60 %) was used as the lipid for the oil phase (10.6 %). The emulsion samples were usually incubated at 50 °C to accelerate lipid oxidation. Lipid oxidation indicators were lipid hydroperoxides and headspace hexanal, determined by solid phase microextraction (SPME) combined with gas chromatography (GC). WPI- or NaCas-stabilised emulsions were prepared using a wide range of protein concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 7.0 or 10.0 %) at two droplet sizes (d32 = 0.31 and 0.65 µm). In general, higher lipid oxidation levels were found for the larger droplet size. Increasing protein concentration led to a decrease in the lipid oxidation rate. The greatest decrease in lipid hydroperoxide levels (values after 4 h) occurred at up to 4.0 % protein concentration. The greatest decrease in hexanal levels (values after 24 h) occurred at up to 4.0 % protein concentration in WPI emulsions (0.31 µm). The hexanal levels were more independent of the protein concentration in the other emulsion types. The hexanal level decreased at protein concentrations > 4.0 % in NaCas emulsions (0.31 and 0.65 µm) and at protein concentrations > 7.0 % in WPI emulsions (0.65 µm). The difference between lipid hydroperoxide generation in emulsions with small and large droplet sizes decreased with increasing protein concentration. This effect was more pronounced in NaCas emulsions. In general, NaCas was a better inhibitor of lipid oxidation than WPI, but WPI appeared to be the better antioxidant at some droplet size/protein concentration combinations. The protein in the continuous phase, i.e. the unadsorbed protein, played an important role in lipid oxidation. In principal, the lipid hydroperoxide and hexanal levels showed the same development over the continuous phase protein concentration as over the protein concentration in WPI and NaCas emulsions (d32 = 0.31 µm). A low NaCas level in the continuous phase already led to a relatively low hexanal level, whereas a higher WPI level was required. When NaCas solution was added to a WPI emulsion or WPI solution was added to a NaCas emulsion, a synergistic antioxidative effect was observed. The high molecular weight fractions (molecular weight = 12000-14000) of WPI and NaCas contained pro-oxidative metal ions that contributed to lipid oxidation in the emulsions. An enrichment of NaCas emulsions with the low molecular weight fraction of NaCas (with a molecular weight = 12000-14000) notably inhibited lipid oxidation. An enrichment of WPI emulsions with the low molecular weight fraction of WPI (with a molecular weight = 12000-14000) also seemed to inhibit lipid oxidation, but the effect was not significant. The protein solutions were enriched with these fractions before emulsion preparation. Pure WPI solution or mixed WPI/NaCas (1:1, weight/weight) solution with 1.12 or 2.24 % protein concentration was heated at 84 °C for up to 40 min, cooled and then used to prepare emulsions. Lipid oxidation was generally not affected by the heat treatment or the degree of whey protein denaturation. However, at the lower WPI concentration, more hexanal was produced for the longer heating times (20, 30 and 40 min) and this appeared to be connected with the physical instability of the emulsions. Greater oxidative stability was found at the higher protein concentration and when the proteins were mixed, pointing to a possible synergistic antioxidative effect of WPI and NaCas. The addition of the free radical source 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH) greatly increased the oxygen uptake and the generation of lipid hydroperoxides in the emulsions. The oxidative stability increased with increasing protein concentration (1.0, 4.0 and 7.0 %). NaCas had a greater antioxidative effect than WPI. The inhibition of oxygen uptake appeared to be largely influenced by the free-radical-scavenging activity of the system, determined by the protein type and the protein concentration, as the radicals were produced linearly over time and oxygen was consumed linearly over time. It can therefore be concluded that free-radical-scavenging activity represents a major antioxidative mechanism of the milk proteins. Oxygen was consumed much faster in emulsions than in protein solutions when the same level of AAPH was incorporated. In a WPI (1.0 % protein) emulsion, much lower levels of protein hydroperoxides than of lipid hydroperoxides developed. This pointed to a much greater reactivity of linoleic acid than of the milk proteins with oxygen. In contrast, the exposure of WPI to oxidising linoleic acid in an emulsion (1.0 % protein) or to AAPH in aqueous solution led to oxidative damage of the whey proteins, indicated by the loss of amino acids. The loss of specific amino acids was different for proteins in the continuous phase or cream phase of an emulsion or in WPI solution. The present study confirms the antioxidative potential of WPI and NaCas and gives new insights into their functionality as oxidative stabilisers in O/W emulsions.

The present study was aimed at extending our knowledge of the antioxidative properties of the milk protein products, whey protein isolate (WPI) and sodium caseinate (NaCas), in oil-in-water (O/W) emulsions rich in polyunsaturated fatty acids (PUFAs). In particular, the objective was to contribute to our understanding of the compositional and processing factors that influence the oxidative stability of protein-stabilised O/W emulsions. Linoleic acid (approximately 60 %) was used as the lipid for the oil phase (10.6 %). The emulsion samples were usually incubated at 50 °C to accelerate lipid oxidation. Lipid oxidation indicators were lipid hydroperoxides and headspace hexanal, determined by solid phase microextraction (SPME) combined with gas chromatography (GC). WPI- or NaCas-stabilised emulsions were prepared using a wide range of protein concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 7.0 or 10.0 %) at two droplet sizes (d32 = 0.31 and 0.65 µm). In general, higher lipid oxidation levels were found for the larger droplet size. Increasing protein concentration led to a decrease in the lipid oxidation rate. The greatest decrease in lipid hydroperoxide levels (values after 4 h) occurred at up to 4.0 % protein concentration. The greatest decrease in hexanal levels (values after 24 h) occurred at up to 4.0 % protein concentration in WPI emulsions (0.31 µm). The hexanal levels were more independent of the protein concentration in the other emulsion types. The hexanal level decreased at protein concentrations > 4.0 % in NaCas emulsions (0.31 and 0.65 µm) and at protein concentrations > 7.0 % in WPI emulsions (0.65 µm). The difference between lipid hydroperoxide generation in emulsions with small and large droplet sizes decreased with increasing protein concentration. This effect was more pronounced in NaCas emulsions. In general, NaCas was a better inhibitor of lipid oxidation than WPI, but WPI appeared to be the better antioxidant at some droplet size/protein concentration combinations. The protein in the continuous phase, i.e. the unadsorbed protein, played an important role in lipid oxidation. In principal, the lipid hydroperoxide and hexanal levels showed the same development over the continuous phase protein concentration as over the protein concentration in WPI and NaCas emulsions (d32 = 0.31 µm). A low NaCas level in the continuous phase already led to a relatively low hexanal level, whereas a higher WPI level was required. When NaCas solution was added to a WPI emulsion or WPI solution was added to a NaCas emulsion, a synergistic antioxidative effect was observed. The high molecular weight fractions (molecular weight = 12000-14000) of WPI and NaCas contained pro-oxidative metal ions that contributed to lipid oxidation in the emulsions. An enrichment of NaCas emulsions with the low molecular weight fraction of NaCas (with a molecular weight = 12000-14000) notably inhibited lipid oxidation. An enrichment of WPI emulsions with the low molecular weight fraction of WPI (with a molecular weight = 12000-14000) also seemed to inhibit lipid oxidation, but the effect was not significant. The protein solutions were enriched with these fractions before emulsion preparation. Pure WPI solution or mixed WPI/NaCas (1:1, weight/weight) solution with 1.12 or 2.24 % protein concentration was heated at 84 °C for up to 40 min, cooled and then used to prepare emulsions. Lipid oxidation was generally not affected by the heat treatment or the degree of whey protein denaturation. However, at the lower WPI concentration, more hexanal was produced for the longer heating times (20, 30 and 40 min) and this appeared to be connected with the physical instability of the emulsions. Greater oxidative stability was found at the higher protein concentration and when the proteins were mixed, pointing to a possible synergistic antioxidative effect of WPI and NaCas. The addition of the free radical source 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH) greatly increased the oxygen uptake and the generation of lipid hydroperoxides in the emulsions. The oxidative stability increased with increasing protein concentration (1.0, 4.0 and 7.0 %). NaCas had a greater antioxidative effect than WPI. The inhibition of oxygen uptake appeared to be largely influenced by the free-radical-scavenging activity of the system, determined by the protein type and the protein concentration, as the radicals were produced linearly over time and oxygen was consumed linearly over time. It can therefore be concluded that free-radical-scavenging activity represents a major antioxidative mechanism of the milk proteins. Oxygen was consumed much faster in emulsions than in protein solutions when the same level of AAPH was incorporated. In a WPI (1.0 % protein) emulsion, much lower levels of protein hydroperoxides than of lipid hydroperoxides developed. This pointed to a much greater reactivity of linoleic acid than of the milk proteins with oxygen. In contrast, the exposure of WPI to oxidising linoleic acid in an emulsion (1.0 % protein) or to AAPH in aqueous solution led to oxidative damage of the whey proteins, indicated by the loss of amino acids. The loss of specific amino acids was different for proteins in the continuous phase or cream phase of an emulsion or in WPI solution. The present study confirms the antioxidative potential of WPI and NaCas and gives new insights into their functionality as oxidative stabilisers in O/W emulsions.

The present study was aimed at extending our knowledge of the antioxidative properties of the milk protein products, whey protein isolate (WPI) and sodium caseinate (NaCas), in oil-in-water (O/W) emulsions rich in polyunsaturated fatty acids (PUFAs). In particular, the objective was to contribute to our understanding of the compositional and processing factors that influence the oxidative stability of protein-stabilised O/W emulsions. Linoleic acid (approximately 60 %) was used as the lipid for the oil phase (10.6 %). The emulsion samples were usually incubated at 50 °C to accelerate lipid oxidation. Lipid oxidation indicators were lipid hydroperoxides and headspace hexanal, determined by solid phase microextraction (SPME) combined with gas chromatography (GC). WPI- or NaCas-stabilised emulsions were prepared using a wide range of protein concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 7.0 or 10.0 %) at two droplet sizes (d32 = 0.31 and 0.65 µm). In general, higher lipid oxidation levels were found for the larger droplet size. Increasing protein concentration led to a decrease in the lipid oxidation rate. The greatest decrease in lipid hydroperoxide levels (values after 4 h) occurred at up to 4.0 % protein concentration. The greatest decrease in hexanal levels (values after 24 h) occurred at up to 4.0 % protein concentration in WPI emulsions (0.31 µm). The hexanal levels were more independent of the protein concentration in the other emulsion types. The hexanal level decreased at protein concentrations > 4.0 % in NaCas emulsions (0.31 and 0.65 µm) and at protein concentrations > 7.0 % in WPI emulsions (0.65 µm). The difference between lipid hydroperoxide generation in emulsions with small and large droplet sizes decreased with increasing protein concentration. This effect was more pronounced in NaCas emulsions. In general, NaCas was a better inhibitor of lipid oxidation than WPI, but WPI appeared to be the better antioxidant at some droplet size/protein concentration combinations. The protein in the continuous phase, i.e. the unadsorbed protein, played an important role in lipid oxidation. In principal, the lipid hydroperoxide and hexanal levels showed the same development over the continuous phase protein concentration as over the protein concentration in WPI and NaCas emulsions (d32 = 0.31 µm). A low NaCas level in the continuous phase already led to a relatively low hexanal level, whereas a higher WPI level was required. When NaCas solution was added to a WPI emulsion or WPI solution was added to a NaCas emulsion, a synergistic antioxidative effect was observed. The high molecular weight fractions (molecular weight = 12000-14000) of WPI and NaCas contained pro-oxidative metal ions that contributed to lipid oxidation in the emulsions. An enrichment of NaCas emulsions with the low molecular weight fraction of NaCas (with a molecular weight = 12000-14000) notably inhibited lipid oxidation. An enrichment of WPI emulsions with the low molecular weight fraction of WPI (with a molecular weight = 12000-14000) also seemed to inhibit lipid oxidation, but the effect was not significant. The protein solutions were enriched with these fractions before emulsion preparation. Pure WPI solution or mixed WPI/NaCas (1:1, weight/weight) solution with 1.12 or 2.24 % protein concentration was heated at 84 °C for up to 40 min, cooled and then used to prepare emulsions. Lipid oxidation was generally not affected by the heat treatment or the degree of whey protein denaturation. However, at the lower WPI concentration, more hexanal was produced for the longer heating times (20, 30 and 40 min) and this appeared to be connected with the physical instability of the emulsions. Greater oxidative stability was found at the higher protein concentration and when the proteins were mixed, pointing to a possible synergistic antioxidative effect of WPI and NaCas. The addition of the free radical source 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH) greatly increased the oxygen uptake and the generation of lipid hydroperoxides in the emulsions. The oxidative stability increased with increasing protein concentration (1.0, 4.0 and 7.0 %). NaCas had a greater antioxidative effect than WPI. The inhibition of oxygen uptake appeared to be largely influenced by the free-radical-scavenging activity of the system, determined by the protein type and the protein concentration, as the radicals were produced linearly over time and oxygen was consumed linearly over time. It can therefore be concluded that free-radical-scavenging activity represents a major antioxidative mechanism of the milk proteins. Oxygen was consumed much faster in emulsions than in protein solutions when the same level of AAPH was incorporated. In a WPI (1.0 % protein) emulsion, much lower levels of protein hydroperoxides than of lipid hydroperoxides developed. This pointed to a much greater reactivity of linoleic acid than of the milk proteins with oxygen. In contrast, the exposure of WPI to oxidising linoleic acid in an emulsion (1.0 % protein) or to AAPH in aqueous solution led to oxidative damage of the whey proteins, indicated by the loss of amino acids. The loss of specific amino acids was different for proteins in the continuous phase or cream phase of an emulsion or in WPI solution. The present study confirms the antioxidative potential of WPI and NaCas and gives new insights into their functionality as oxidative stabilisers in O/W emulsions.

The present study was aimed at extending our knowledge of the antioxidative properties of the milk protein products, whey protein isolate (WPI) and sodium caseinate (NaCas), in oil-in-water (O/W) emulsions rich in polyunsaturated fatty acids (PUFAs). In particular, the objective was to contribute to our understanding of the compositional and processing factors that influence the oxidative stability of protein-stabilised O/W emulsions. Linoleic acid (approximately 60 %) was used as the lipid for the oil phase (10.6 %). The emulsion samples were usually incubated at 50 °C to accelerate lipid oxidation. Lipid oxidation indicators were lipid hydroperoxides and headspace hexanal, determined by solid phase microextraction (SPME) combined with gas chromatography (GC). WPI- or NaCas-stabilised emulsions were prepared using a wide range of protein concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 7.0 or 10.0 %) at two droplet sizes (d32 = 0.31 and 0.65 µm). In general, higher lipid oxidation levels were found for the larger droplet size. Increasing protein concentration led to a decrease in the lipid oxidation rate. The greatest decrease in lipid hydroperoxide levels (values after 4 h) occurred at up to 4.0 % protein concentration. The greatest decrease in hexanal levels (values after 24 h) occurred at up to 4.0 % protein concentration in WPI emulsions (0.31 µm). The hexanal levels were more independent of the protein concentration in the other emulsion types. The hexanal level decreased at protein concentrations > 4.0 % in NaCas emulsions (0.31 and 0.65 µm) and at protein concentrations > 7.0 % in WPI emulsions (0.65 µm). The difference between lipid hydroperoxide generation in emulsions with small and large droplet sizes decreased with increasing protein concentration. This effect was more pronounced in NaCas emulsions. In general, NaCas was a better inhibitor of lipid oxidation than WPI, but WPI appeared to be the better antioxidant at some droplet size/protein concentration combinations. The protein in the continuous phase, i.e. the unadsorbed protein, played an important role in lipid oxidation. In principal, the lipid hydroperoxide and hexanal levels showed the same development over the continuous phase protein concentration as over the protein concentration in WPI and NaCas emulsions (d32 = 0.31 µm). A low NaCas level in the continuous phase already led to a relatively low hexanal level, whereas a higher WPI level was required. When NaCas solution was added to a WPI emulsion or WPI solution was added to a NaCas emulsion, a synergistic antioxidative effect was observed. The high molecular weight fractions (molecular weight = 12000-14000) of WPI and NaCas contained pro-oxidative metal ions that contributed to lipid oxidation in the emulsions. An enrichment of NaCas emulsions with the low molecular weight fraction of NaCas (with a molecular weight = 12000-14000) notably inhibited lipid oxidation. An enrichment of WPI emulsions with the low molecular weight fraction of WPI (with a molecular weight = 12000-14000) also seemed to inhibit lipid oxidation, but the effect was not significant. The protein solutions were enriched with these fractions before emulsion preparation. Pure WPI solution or mixed WPI/NaCas (1:1, weight/weight) solution with 1.12 or 2.24 % protein concentration was heated at 84 °C for up to 40 min, cooled and then used to prepare emulsions. Lipid oxidation was generally not affected by the heat treatment or the degree of whey protein denaturation. However, at the lower WPI concentration, more hexanal was produced for the longer heating times (20, 30 and 40 min) and this appeared to be connected with the physical instability of the emulsions. Greater oxidative stability was found at the higher protein concentration and when the proteins were mixed, pointing to a possible synergistic antioxidative effect of WPI and NaCas. The addition of the free radical source 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH) greatly increased the oxygen uptake and the generation of lipid hydroperoxides in the emulsions. The oxidative stability increased with increasing protein concentration (1.0, 4.0 and 7.0 %). NaCas had a greater antioxidative effect than WPI. The inhibition of oxygen uptake appeared to be largely influenced by the free-radical-scavenging activity of the system, determined by the protein type and the protein concentration, as the radicals were produced linearly over time and oxygen was consumed linearly over time. It can therefore be concluded that free-radical-scavenging activity represents a major antioxidative mechanism of the milk proteins. Oxygen was consumed much faster in emulsions than in protein solutions when the same level of AAPH was incorporated. In a WPI (1.0 % protein) emulsion, much lower levels of protein hydroperoxides than of lipid hydroperoxides developed. This pointed to a much greater reactivity of linoleic acid than of the milk proteins with oxygen. In contrast, the exposure of WPI to oxidising linoleic acid in an emulsion (1.0 % protein) or to AAPH in aqueous solution led to oxidative damage of the whey proteins, indicated by the loss of amino acids. The loss of specific amino acids was different for proteins in the continuous phase or cream phase of an emulsion or in WPI solution. The present study confirms the antioxidative potential of WPI and NaCas and gives new insights into their functionality as oxidative stabilisers in O/W emulsions.

The present study was aimed at extending our knowledge of the antioxidative properties of the milk protein products, whey protein isolate (WPI) and sodium caseinate (NaCas), in oil-in-water (O/W) emulsions rich in polyunsaturated fatty acids (PUFAs). In particular, the objective was to contribute to our understanding of the compositional and processing factors that influence the oxidative stability of protein-stabilised O/W emulsions. Linoleic acid (approximately 60 %) was used as the lipid for the oil phase (10.6 %). The emulsion samples were usually incubated at 50 °C to accelerate lipid oxidation. Lipid oxidation indicators were lipid hydroperoxides and headspace hexanal, determined by solid phase microextraction (SPME) combined with gas chromatography (GC). WPI- or NaCas-stabilised emulsions were prepared using a wide range of protein concentrations (0.5, 1.0, 2.0, 3.0, 4.0, 7.0 or 10.0 %) at two droplet sizes (d32 = 0.31 and 0.65 µm). In general, higher lipid oxidation levels were found for the larger droplet size. Increasing protein concentration led to a decrease in the lipid oxidation rate. The greatest decrease in lipid hydroperoxide levels (values after 4 h) occurred at up to 4.0 % protein concentration. The greatest decrease in hexanal levels (values after 24 h) occurred at up to 4.0 % protein concentration in WPI emulsions (0.31 µm). The hexanal levels were more independent of the protein concentration in the other emulsion types. The hexanal level decreased at protein concentrations > 4.0 % in NaCas emulsions (0.31 and 0.65 µm) and at protein concentrations > 7.0 % in WPI emulsions (0.65 µm). The difference between lipid hydroperoxide generation in emulsions with small and large droplet sizes decreased with increasing protein concentration. This effect was more pronounced in NaCas emulsions. In general, NaCas was a better inhibitor of lipid oxidation than WPI, but WPI appeared to be the better antioxidant at some droplet size/protein concentration combinations. The protein in the continuous phase, i.e. the unadsorbed protein, played an important role in lipid oxidation. In principal, the lipid hydroperoxide and hexanal levels showed the same development over the continuous phase protein concentration as over the protein concentration in WPI and NaCas emulsions (d32 = 0.31 µm). A low NaCas level in the continuous phase already led to a relatively low hexanal level, whereas a higher WPI level was required. When NaCas solution was added to a WPI emulsion or WPI solution was added to a NaCas emulsion, a synergistic antioxidative effect was observed. The high molecular weight fractions (molecular weight = 12000-14000) of WPI and NaCas contained pro-oxidative metal ions that contributed to lipid oxidation in the emulsions. An enrichment of NaCas emulsions with the low molecular weight fraction of NaCas (with a molecular weight = 12000-14000) notably inhibited lipid oxidation. An enrichment of WPI emulsions with the low molecular weight fraction of WPI (with a molecular weight = 12000-14000) also seemed to inhibit lipid oxidation, but the effect was not significant. The protein solutions were enriched with these fractions before emulsion preparation. Pure WPI solution or mixed WPI/NaCas (1:1, weight/weight) solution with 1.12 or 2.24 % protein concentration was heated at 84 °C for up to 40 min, cooled and then used to prepare emulsions. Lipid oxidation was generally not affected by the heat treatment or the degree of whey protein denaturation. However, at the lower WPI concentration, more hexanal was produced for the longer heating times (20, 30 and 40 min) and this appeared to be connected with the physical instability of the emulsions. Greater oxidative stability was found at the higher protein concentration and when the proteins were mixed, pointing to a possible synergistic antioxidative effect of WPI and NaCas. The addition of the free radical source 2,2’-azobis(2-amidinopropane) dihydrochloride (AAPH) greatly increased the oxygen uptake and the generation of lipid hydroperoxides in the emulsions. The oxidative stability increased with increasing protein concentration (1.0, 4.0 and 7.0 %). NaCas had a greater antioxidative effect than WPI. The inhibition of oxygen uptake appeared to be largely influenced by the free-radical-scavenging activity of the system, determined by the protein type and the protein concentration, as the radicals were produced linearly over time and oxygen was consumed linearly over time. It can therefore be concluded that free-radical-scavenging activity represents a major antioxidative mechanism of the milk proteins. Oxygen was consumed much faster in emulsions than in protein solutions when the same level of AAPH was incorporated. In a WPI (1.0 % protein) emulsion, much lower levels of protein hydroperoxides than of lipid hydroperoxides developed. This pointed to a much greater reactivity of linoleic acid than of the milk proteins with oxygen. In contrast, the exposure of WPI to oxidising linoleic acid in an emulsion (1.0 % protein) or to AAPH in aqueous solution led to oxidative damage of the whey proteins, indicated by the loss of amino acids. The loss of specific amino acids was different for proteins in the continuous phase or cream phase of an emulsion or in WPI solution. The present study confirms the antioxidative potential of WPI and NaCas and gives new insights into their functionality as oxidative stabilisers in O/W emulsions.