Effect of Dietary Fat and Vitamin E on Colour Stability ... - Science Direct

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Meaf .Science, Vol. 48, No. 3/4, 301-318, 1998 0 1998 Elsevier Science Ltd All rights reserved. Printed in Great Britain 0309-1740/98 $19.00+0.00

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Effect of Dietary Fat and Vitamin E on Colour Stability and on Lipid and Protein Oxidation in Turkey Meat During Storage Y. Mercier,a P. Gatellier,” M. Viau,b H. Remignon” & M. Renerrea* “INRA, Station de Recherches sur la Viande, 63122 St Genes Champanelle, bINRA, Leima, BP 1627,44316 Nantes Cedex 03, France ‘INRA, Station de Recherches Avicoles, 37380 Nouzilly, France

France

(Received 18 March 1997; revised version received 28 September 1997; accepted 30 September 1997)

ABSTRACT The objectives of the study were to investigate the effects of dietary fat (6% soya oil or rapeseed oil or tallow), together with tocopheryl acetate at either a basal (30ppm) or a supplemented (400ppm) level for 16 weeks on lipid and protein oxidation, including myoglobin, during refrigerated storage of turkey muscles. When turkeys were fed tallow in particular, vitamin E supplementation improved the vitamin E status of the muscles. Vitamin E supplementation signtjicantly delayed lipid oxidation measured by TBARS, whatever the dietary fat. TBARS were highest in meat from animals fed soya oil. Vitamin E supplementation had no positive effect on colour stability of meat during refrigerated storage. Feeding soya oil induced significantly higher oxidation of proteins (carbonyl content) than rapeseed oil or tallow and vitamin E supplementation induced a slight decrease in carbonyl content at day 9 of storage for M. sartorius. SH content was significantly higher in vitamin E supplemented M. sartorius and M. pectoralis than in controls. 0 1998 Elsevier Science Ltd. AN rights reserved

INTRODUCTION Colour stability is one of the most important quality attributes contributing to meat shelflife and studies have been carried out in our laboratory to explain the deterioration of the bright red colour of beef (Renerre and Labas, 1987a). In France, the consumption of

turkey meat is rapidly increasing and the maintenance of colour and lipid stability of overwrapped meat is important. In comparison to beef, few studies have been carried out on the discoloration of ‘white’ meats such as turkey where the discoloration is characterized by fading from pink/yellow to yellow/brown. Lipid oxidation causes the development of rancidity and ‘warmed-over flavors’ and membrane phospholipids are the sites where oxidative changes are initiated in meat. It is well known that to decrease these oxidative processes, vitamin E is a highly-efficient antioxidant in cell membranes, acting as a chain-breaker. Experiments have shown that *To whom correspondence

should be addressed. Fax: 00 33 73 62 42 68; e-mail: [email protected] 301

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vitamin E reduced lipid oxidation in chicken meat (Sheehy et al., 1993) and pork (Monahan et al., 1992) during storage. In frozen turkey meat, Bartov et al. (1983) and Bartov and Kanner (1996) have shown that dietary vitamin E decreased TBARS values. Many authors have shown that lipid and pigment oxidation are closely related in beef (Faustman et al., 1989; Gatellier et al., 1992; Renerre and Labadie, 1993), pork (Asghar et al., 1991; Monahan et al., 1994) and lamb (Guidera et al., 1997). Moreover, many experiments have indicated that vitamin E supplementation results in greater colour and lipid stability in beef (Faustman et al., 1989; Arnold et al., 1993u), by delaying oxidation of phospholipids. In model systems obtained from beef (mitochondria and microsomes), it was shown that lipid and myoglobin oxidation could be lowered by addition of vitamin E (Anton et al., 1993~). In turkey, although a dietary supplementation in vitamin E may lower lipid oxidation, it is not known if it improves colour stability. The hydroxyl radical OH’, and other oxygenated free-radicals, damage cellular targets such as lipids and myoglobin, and although proteins in muscle cells are also targets for oxygen radicals in vivo, much less is known about radical attacks on proteins (Stadtman, 1987) particularly during meat maturation. Protein oxidation, which occurs in beef during maturation (Mercier et al., 1995; Martinaud et al., 1997) and in fish (Srinivasan and Hultin, 1995), as measured by the total carbonyl content, could be a sensitive indicator of a cell’s exposure to oxygen radicals. It is not established if vitamin E supplementation can lower protein oxidation during meat maturation. Moreover in meat, phospholipid oxidation is dependent on the proportion of polyunsaturated fatty acids (PUFA) in the diet and it is known that dietary fat in non-ruminant animals, such as broilers or turkeys, influence PUFA content and muscle food quality during storage (Marusich et al., 1975; Sklan et al., 1983; Sheldon, 1984; Brandon et al., 1993). The aim of this work was to develop a better understanding of the effects of dietary modification, with different fat sources and vitamin E supplementation, on lipid and protein oxidation, including myoglobin, during a refrigerated storage of M. pectoralis and M. sartorius from turkeys. MATERIALS

AND METHODS

Animals and diets

Seventy-two male turkeys of BUT strain were reared at the Station of Poultry Research (INRA, Nouzilly) and slaughtered at 16 weeks. The animals, which were fed ad libitum, received a common basal diet enriched with 6% of one of the following fat sources: tallow (n = 24), rapeseed oil (n = 24) or soya oil (n = 24). For each diet, animals were divided into two groups according to the vitamin E supplementation (cu-tocopheryl-acetate from Hoffman-Laroche, France): 30 ppm (control animals: C) or 400 ppm (vitamin E supplemented animals: E). After killing and bleeding, the M. pectoralis (P) and M. sartorius (S) were immediately removed from the left side of the carcasses, stored in ice and analysed the day after slaughter for vitamin E content. Muscles from the right side were removed about 6 hr after slaughter and stored at 2°C for 18 hr. Meat samples were transported in ice to the laboratory in Theix and 24 hr after slaughter were placed on fibreboard trays, overwrapped with an oxygen-permeable PVC film (10.000 cm3 m-* 24 hr-‘) and stored in darkness at 3°C for up to nine days.

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Vitamin E content of muscles Vitamin E content was determined according to the method of Buttriss and Diplock (1984). After saponification and hexane extraction, all the samples were analysed by normal phase HPLC (Lichrospher Si 60 column from Merck) fitted with a fluorimeter detector (excitation 292 nm/emission 330nm). The results are expressed as pg vitamin E g-i of tissue. pH and haeminic iron Haeminic iron was determined in 5 g muscle samples, using the Homsey method (1956), and pH was measured (Schott pH meter) directly in the cuts used for colour determination. Colour measurements At 24 hr post mortem, muscles, which were placed on fibreboard trays and overwrapped in oxygen-permeable PVC film, were exposed in darkness to air at 3°C from 2 hr (day 1) to day 9. Reflectance spectra from 360 to 760 nm were obtained with a Uvikon 860 spectrocalorimeter (Kontron) equipped with an integrating sphere and colour coordinates were calculated in the CIELAB system (1976) at days 1, 3 and 9. The results were expressed as lightness (L*), redness (a*), yellowness (b*), hue and chroma. The rates of meat discoloration were determined as R6s0-R5s0 and K/S&K/& (Renerre, 1984). Metmyoglobin content was measured according to Krzywicki (1979) Lipid oxidation Lipid oxidation was measured by the TBARS content according to the method of Lynch and Frei (1993). Samples (0.5 ml) (1 g muscle minced in 10 ml of KC1 O-15 M + BHT 0.1 mM) were incubated with 1% (w/v) 2-thiobarbituric acid in 50 mM NaOH (0.25 ml) and 2.8% (w/v) trichloroacetic acid (0.25 ml) in a boiling water bath for 10 min. After cooling to room temperature, the pink chromogen was extracted into n-butanol (2.0ml) and its absorbance measured at 535nm. TBARS concentrations were calculated using 1,1,3,3tetramethoxypropane (O-8 PM) as standard. The analyses were performed on samples stored 1,3 or 9 days in darkness at 3°C. The results were expressed as mg MDA kg-’ meat. Protein oxidation Carbonyl

content

One gram of muscle was ground with a Waring-blender in lOm1 KC1 (0.15 M) solution. Muscle extract was divided into two equal aliquots containing approximately 0.7-1.0 mg of protein each. Protein was precipitated in both aliquots by 10% trichloracetic acid (w/v, final concentration) and centrifuged for 10 min at 2000 g. One pellet was treated with 2MHCl and the other with an equal volume of 0.2% (w/v) 2,4_dinitrophenylhydrazine (DNPH) in 2 M HCl (Oliver et al., 1987). Both samples were incubated for 1 hr at room temperature and stirred regularly. The samples were reprecipitated with 10% TCA (final concentration) and the pellets washed twice with ethanol:ethyl acetate (1:l). The pellets were carefully drained and dissolved in 6 M guanidine HCl with 20 mM sodium phosphate buffer, pH 6.5. When insoluble fragments were present, they were removed by centrifugation for 10 min at 2000 g. The difference spectrum of the DNPH-treated sample against the HCl control was determined. Protein concentration was calculated at 280 nm in the HCl control using BSA

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in guanidine as standard. The results were expressed as nmol of DNPH incorporated mg-’ of protein based on an average absorption of 21.0 mM-’ cm-’ at 370 nm for protein hydrazones. Thiol content

One gram of muscle was ground in a Waring-blender in lOm1 KC1 (0.15M) solution. Samples were diluted (l/50) in 100mM phosphate buffer pH 8.0 and 8M urea, To 1 ml of protein solution, 10 ~1 of 2,2’-dithio-bis(nitropyridine) (DTNP) was added. Incubation was for 1 hr at room temperature and the absorbance at 386 nm against a blank of protein at the same concentration without DTNP determined (Winterbourn, 1990). Absorbance of DTNP alone at 386nm was subtracted and the protein concentration measured by Biuret method (Gornal et al, 1949). The results were expressed as nmol of free thiol mgg’ protein using an extinction coefficient of 14.0 mM-’ cm-‘. Statistical analysis

Data were analyzed at day 9 by a two-way analysis of variance (SAS) to assess the effects of fat and/or vitamin E dietary supplementation. Mean values were compared using the Newman-Keuls test. RESULTS

AND DISCUSSION

Vitamin E content

In muscles, vitamin E content was significantly (p < 0.05) affected by the vitamin E level in the diet (Table 1). In M. pectoralis and M. sartorius, vitamin E content of supplemented animals was almost 6 times greater than in the controls. M. sartorius contained about twice as much vitamin E as M. pectoralis whatever the dietary level of vitamin E (p tallow > rapeseed). The differences are unexpected but it is possible that during aging, oxidation is low and differences between treatments is difficult to measure. However two-way variance analysis for M. sartorius showed a significant interaction between fat and vitamin. At day 9, it was noted that -SH content was significantly higher (p < 0.05) in supplemented animals than in control ones: (118/100 nmol mg-’ protein) for M. sartorius and (104/96nmolmg-i protein) for M. pectoralis [Tables 3(b) and 5(b)]. There was also a significant fat/vitamin interaction for M. sartorius (Table 6). Conversely, with time, and whatever the treatment, a small decrease in thiol groups was noted indicating low oxidation of proteins during storage. The decrease in -SH groups was greater in control animals than in supplemented ones. The differences in -SH content between day 1 and day 9 were more pronounced in M. sartorius than in M. pectoralis (results not shown). These results were also more pronounced with birds fed soya oil and tallow than with those fed rapeseed oil.

CONCLUSION When turkey meat was stored for 1, 3 or 9 days, vitamin E supplementation significantly delayed lipid oxidation as measured by TBARS, whatever the dietary fat. Vitamin E supplementation had no positive effect either on initial colour or colour stability regardless of diet. At day 9, for M. sartorius, feeding with soya oil induced significantly higher oxidation of proteins and supplementation with vitamin E resulted in a significant decrease in carbonyl content. At day 9, for both muscles, SH content was significantly higher in supplemented than in control birds.

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ACKNOWLEDGEMENTS The authors wish to express their gratitude English writing style. This work was financed

to Dr E. Dransfield by AIR programme

for the corrections in from EEC (DG XII).

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