Els Vossen

Effect of rosemary extract dose on lipid oxidation, colour stability and antioxidant concentrations, in reduced nitrite liver pâtés

The oxidative stability of liver pâté was investigated in relation to different doses of rosemary extract (RE) and sodium nitrite. Colour stability, lipid oxidation (TBARS) and concentrations of ascorbic acid, α-tocopherol, carnosic acid and nitrite were measured on the batters before cooking and on the cooked liver pâté before and after exposure to light and air for 48 h at 4°C. Results showed that the use of RE significantly reduced lipid oxidation, whereas it had no effect on colour stability. Ascorbic acid and nitrite concentrations were significantly higher and lower respectively when RE was added. RE dose-dependently increased the concentration of carnosic acid. Lower sodium nitrite doses resulted in significantly lower nitrite concentrations and slightly lower TBARS values. It was concluded that in liver pâté sodium nitrite levels may be lowered to 80 mg/kg without negatively affecting colour and lipid stability and that the use of RE may help in maintaining lipid stability.

Nitrite curing of chicken, pork, and beef inhibits oxidation but does not affect N‑nitroso compound (NOC)-specific DNA adduct formation during in vitro digestion

Uncured and nitrite-cured chicken, pork, and beef were used as low, medium, and high sources of heme-Fe, respectively, and exposed to an in vitro digestion model simulating the mouth, stomach, duodenum, and colon. With increasing content of iron compounds, up to 25-fold higher concentrations of the toxic lipid oxidation products malondialdehyde, 4-hydroxy-2-nonenal, and other volatile aldehydes were formed during digestion, together with increased protein carbonyl compounds as measurement of protein oxidation. Nitrite curing of all meats lowered lipid and protein oxidation to the level of oxidation in uncured chicken. Strongly depending on the individual fecal inoculum, colonic digestion of beef resulted in significantly higher concentrations of the NOC-specific DNA adduct O(6)-carboxymethyl-guanine compared to chicken and pork, whereas nitrite curing had no significant effect. This study confirms previously reported evidence that heme-Fe is involved in the epidemiological association between red meat consumption and colorectal cancer, but questions the role of nitrite curing in this association.

Dog rose (Rosa canina L.) as a functional ingredient in porcine frankfurters without added sodium ascorbate and sodium nitrite.

The effect of dog rose (Rosa canina L.; RC), rich in polyphenols and ascorbic acid, on lipid and protein oxidation, colour stability and texture of frankfurters was investigated. Four treatments were prepared: with 5 or 30 g/kg RC extract and without sodium ascorbate and sodium nitrite (5RC and 30RC, respectively), a positive control (with sodium ascorbate and sodium nitrite; PC) and a negative control (without sodium ascorbate, sodium nitrite or RC extract; NC). Hexanal values were much higher throughout storage in NC compared to RC and PC frankfurters (P<0.001). The RC extracts protected against protein oxidation, but not as efficiently as PC (P<0.05). In the RC treated frankfurters, lower a* values were measured compared to PC due to the lack of sodium nitrite. In conclusion, dog rose can act as a natural antioxidant in frankfurters, but not as full replacer for sodium nitrite.

Effect of dietary antioxidant supplementation on the oxidative status of plasma in broilers

In this study, the effect of dietary antioxidants on the plasma oxidative status of growing birds fed a diet rich in polyunsaturated fatty acids was investigated. One-day-old broilers were fed for 42 days a diet containing 4% linseed oil and supplemented with single plant extracts rich in antioxidants (natural tocopherols, rosemary, grape seed, green tea, tomato) or a combination of some of these plant extracts, in two different total doses (100 and 200 mg product/kg feed). A diet with synthetic antioxidants with and without α-tocopheryl acetate (200 mg/kg feed) were also included. The plasma oxidative status was evaluated measuring the ferric reducing ability of plasma (FRAP), the superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity. Lipid peroxidation was measured by thiobarbituric acid-reactive substances (TBARS). No significant effect of the dietary treatments was observed for FRAP as well as for TBARS. However, diet affected GSH-Px activity (p = 0.002) and a trend for an effect on SOD activity was observed (p=0.084). A higher GSH-Px activity was found for 200 mg/kg tomato extract and natural α-tocopherol in relation to the corresponding 100 mg/kg treatment, and the lowest GSH-Px activity was measured for the synthetic antioxidants treatment. The lowest and highest SOD activity were found for the 200. and 100 mg/kg treatment with tomato extract respectively. In conclusion, the oxidative status and lipid oxidation of plasma in broilers was not affected by feeding natural antioxidant extracts at the doses in the present study, but some changes in antioxidant enzyme activities were observed, of which the implication remains to be elucidated.

Effect of sodium ascorbate dose on the shelf life stability of reduced nitrite liver pâtés

The effect of sodium ascorbate (SA; 500, 750, 1000 mg/kg) and sodium nitrite (SN; 40, 80, 120 mg/kg) doses on the shelf-life stability of liver pâtés was investigated in a full factorial design. Clear dose-dependent responses of the added SN or SA were found for the concentrations of nitrite, ascorbic acid and dehydroascorbic acid in the raw batters and in the cooked pâtés before and after 48 h of chilled display. Decreasing the SN dose to 80 mg/kg had no negative impact on the colour stability (a* value) and lipid oxidation (TBARS), and no additional antioxidant effect of SA was noticed. Lowering SN to 40 mg/kg resulted in proper colour formation, but the colour stability was inferior and lipid oxidation increased. Yet, increasing the amount of SA, at this low SN dose, resulted in lower TBARS values. Decreasing the SN dose to 80 or 40 mg/kg had no distinct effect on protein oxidation, which was however only measured by carbonyl content.

Increased oxidative and nitrosative reactions during digestion could contribute to the association between well-done red meat consumption and colorectal cancer

Uncured and nitrite-cured pork were subjected, raw, cooked (65 °C, 15 min) or overcooked (90 °C, 30 min), to an in vitro digestion model, which includes mouth, stomach, duodenum, and colon phases. Heating of uncured meat resulted in a pronounced increase in lipid and protein oxidation products throughout digestion. Nitrite-curing had an antioxidant effect during digestion, but this effect disappeared when the meat was overcooked, resulting in up to ninefold higher 4-hydroxy-2-nonenal concentrations compared with digested nitrite-cured raw and cooked pork. Colonic digesta contained significantly higher concentrations of the NOC-specific DNA adduct O(6)-carboxy-methylguanine when pork underwent a more intense heating procedure, independent of nitrite-curing, depending strongly on the fecal inoculum used. Since processed meats are usually nitrite-cured, the present study suggests that overcooking processed meat is likely to result in the formation of genotoxic compounds during digestion and should, therefore, be avoided.

Protein oxidation affects proteolysis in a meat model system

The effect of hydrogen peroxide-induced protein oxidation and pH (4.8 and 5.2) on meat proteolysis was investigated in a meat model system for dry fermented sausages. In oxidised samples, increased protein carbonyl contents and decreased thiol concentrations were found. The initial concentration of protein carbonyls was significantly lower in oxidised samples at pH4.8 than in ones at pH5.2, but after ten days comparable levels were reached. The inhibition of proteolysis by the addition of a protease inhibitor cocktail did not influence protein oxidation. Yet, proteolysis was negatively affected by low pH values as well as by oxidation, resulting in a reduced release of amino acids during ripening.

Fat content and nitrite-curing influence the formation of oxidation products and NOC-specific DNA adducts during in vitro digestion of meat.

The effects of fat content and nitrite-curing of pork were investigated on the formation of cytotoxic and genotoxic lipid oxidation products (malondialdehyde, 4-hydroxy-2-nonenal, volatile simple aldehydes), protein oxidation products (protein carbonyl compounds) and NOC-specific DNA adducts (O6-carboxy-methylguanine) during in vitro digestion. The formation of these products during digestion is suggested to be responsible for the association between red meat and processed meat consumption and colorectal cancer risk. Digestion of uncured pork to which fat was added (total fat content 5 or 20%), resulted in significantly higher lipid and protein oxidation in the mimicked duodenal and colonic fluids, compared to digestion of pork without added fat (1% fat). A higher fat content also significantly favored the formation of O6-carboxy-methylguanine in the colon. Nitrite-curing of meat resulted in significantly lower lipid and protein oxidation before and after digestion, while an inconsistent effect on the formation of O6-carboxy-methylguanine was observed. The presented results demonstrate that haem-Fe is not solely responsible for oxidation and nitrosation reactions throughout an in vitro digestion approach but its effect is promoted by a higher fat content in meat.

Protein oxidation and protein nitration influenced by sodium nitrite in two different meat model systems.

The effect of NaNO2 on protein oxidation was studied in isolated myofibrillar protein isolates (100 and 1000 mg NaNO2/kg) and in porcine patties (18 and 180 mg NaNO2/kg). In addition, the potential use of 3-nitrotyrosine as a specific marker for reactive nitrogen species mediated nitration was investigated. Overall, no distinct pro- or antioxidant effect of NaNO2 against carbonyl formation was observed in the isolates or in the patties. However, in the isolates, higher protein carbonyl concentrations were found in the NaNO2-treated samples compared to the treatment without added nitrite immediately after the addition of oxidants and NaNO2. Addition of 180 mg NaNO2/kg to patties resulted in significantly lower thiol concentrations at 4 and 7 days of illuminated chilled display compared to the treatments with 0 and 18 mg/kg NaNO2, whereas no effect was observed in the isolates. No effect of NaNO2 was found on the protein solubility of either meat model. 3-Nitrotyrosine was present in all samples, but no clear effect of NaNO2 addition or oxidation time was observed.

Protection of polyunsaturated oils against ruminal biohydrogenation and oxidation during storage using a polyphenol oxidase containing extract from red clover

Polyunsaturated fatty acid (PUFA) are to a large extent subject to biohydrogenation in a ruminal environment, which results to the healthy value of these PUFA being lost upon dietary addition to ruminants. PUFA are also prone to lipid oxidation upon storage. Therefore, it was tested whether emulsions could be protected against in vitro ruminal biohydrogenation and oxidation during storage by using protein extracts rich in polyphenol oxidase, an enzyme responsible for browning of plant tissues. PUFA rich emulsions were made with a protein extract from red clover (Trifolium pratense L.) before adding a synthetic diphenol (4-methylcatechol) to induce protection. Results after in vitro incubation confirmed the hypothesis and indicated the potential to prevent PUFA in linseed or fish oil from ruminal biohydrogenation and oxidation during storage through addition of 4-methylcatechol to the emulsions. Protection depended on the amount of oil present and protein concentrations in the emulsions. Protection efficiency increased with increasing the amounts of diphenol present in the emulsion per unit interfacial surface area. It is suggested that protection is caused by an effective encapsulation by cross-linking of the protein layer at the emulsion interface. For the first time, a method is described to protect PUFA using an enzyme abundantly available in nature, polyphenol oxidase, in combination with 4-methylcatechol.