Inhibition of hemoglobin-mediated lipid oxidation in washed fish muscle by cranberry components

Fractions enriched in phenolic acids (Fraction 1), anthocyanins (Fraction 2), flavonols (Fractions 3 and 4) and proanthocyanidins (Fractions 5 and 6) were prepared from cranberry powder using Sephadex LH-20 chromatography. Fractions 2, 3, 4, and 5 had nearly equivalent reactivity in the total phenolate assay employed per mg dry weight of each fraction while Fractions 1 and 6 were less reactive. The ability of cranberry fractions to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals as well as their inhibitory effects on hemoglobin (Hb)-mediated lipid oxidation in washed cod muscle were assessed. Addition of cranberry fractions at a level of 74 μmol quercetin equivalents per kg of washed cod muscle extended the induction time of thiobarbituric acid reactive substances (TBARS) formation in the order: Fraction 1, Fraction 3, Fraction 4 > Fraction 2 > Fraction 5 > Fraction 6. This suggests that oligomeric polyphenols (e.g., proanthocyanidins) were least effective at inhibiting Hb-mediated lipid oxidation in washed cod muscle compared to the other classes of polyphenolics in cranberry. The ability of the different cranberry fractions to scavenge DPPH radicals did not reflect their relative ability to inhibit lipid oxidation in the washed cod muscle system. Quercetin was tentatively identified as a component in cranberry that was especially effective at inhibiting Hb-mediated lipid oxidation. The ability of flavonol and proanthocyanidin-enriched fractions to inhibit Hb-mediated lipid oxidation in spite of efforts to wash away the added polyphenolics prior to Hb addition indicated these classes of polyphenolics had binding affinities for insoluble components of washed cod muscle.     

Characteristics of myoglobin and haemoglobin-mediated lipid oxidation in washed mince from bighead carp (Hypophthalmichthys nobilis)

Myoglobin (Mb) and haemoglobin (Hb) accounted for 61% and 39% of the total haem-protein in bighead carp (Hypophthalmichthys nobilis) dark muscle, respectively. Molecular weight of Mb and monomeric-Hb was ∼16 kDa. Haemin loss from metHb was more rapid, compared to metMb (pH 6.0, 4 °C). Pro-oxidative activities of oxyMb/Hb and metMb/Hb were examined in washed mince during 9 days of iced storage (pH 6.0). Soret measurements suggested the existence of holoMb throughout storage. For Hb, weakening of haem-globin linkage was observed, especially for metHb which had undetectable Soret after 3 days of storage. Loss of redness was more rapid and extensive in washed mince containing Hb, compared to Mb. During storage, Hb induced larger amounts of peroxides, thiobarbituric acid-reactive substances and hexanal than did Mb (p < 0.05), especially for met-form. Thus, Hb had lower haemin affinity and was a stronger pro-oxidant than Mb.     

Retardation of myoglobin and haemoglobin-mediated lipid oxidation in washed bighead carp by phenolic compounds

Antioxidative activities of phenolic compounds (caffeic acid, gallic acid and tannic acid; 200 ppm) in washed mince (pH 6), with added myoglobin (Mb) and haemoglobin (Hb), from bighead carp (Hypophthalmichthys nobilis), during 9 days of iced storage, were studied. Tannic acid exhibited the preventive effect on discolouration of washed mince containing Mb or Hb during storage (P < 0.05). High peroxide value (PV) was found and large amount of, thiobarbituric acid-reactive substances (TBARS) and hexanal were formed in washed mince containing haem proteins, especially Hb. As determined by apo Streptococcal haem-associated protein, Hb had the lower haem affinity than Mb. Phenolic compounds, especially caffeic acid and gallic acid, could lower lipid oxidation induced by Mb or Hb throughout storage (P < 0.05). Prevention of haem release, as well as inhibition of lipid oxidation induced by haem proteins with selected phenolic compounds, should be an alternative means in lowering discolouration and lipid oxidation in fish muscle.     

Effect of caffeic acid on haemoglobin‐mediated lipid and protein oxidation in washed cod mince during ice and frozen storage

BACKGROUND: Little is known about the relation between haemoglobin (Hb)‐mediated lipid and protein oxidation in muscle foods and how these two reactions can be inhibited by naturally occurring antioxidants. This study was aimed at evaluating (1) lipid oxidation and protein oxidation induced by 20 µmol L^?1 Hb during chilled and frozen storage of washed cod mince and (2) the efficiency of 10–1000 ppm (0.063–6.3 mmol L^?1) caffeic acid in preventing these reactions. RESULTS: Addition of 20 µmol L^?1 Hb increased peroxide value (PV), rancid odour, protein carbonylation, protein insolubilisation, redness loss and α‐tocopherol loss in ice‐stored washed cod mince. Since both lipid and protein oxidation developed at the same time, it was not possible to conclude which reaction initiated the other. All studied reactions were efficiently inhibited by ≥ 50 ppm caffeic acid, which could be a result of α‐tocopherol regeneration, general radical scavenging, reduced formation of oxidised Hb forms and/or conformational changes in Hb structure. During frozen storage the only clear effect of Hb was increased PV, and here caffeic acid was less efficient as an antioxidant. CONCLUSION: Hb‐induced lipid and protein oxidation occurred quickly in ice‐stored washed cod mince, and the two reactions could not be separated in time. During frozen storage, Hb caused only limited lipid oxidation. Caffeic acid (≥50 ppm) was an efficient antioxidant during ice storage. Copyright © 2010 Society of Chemical Industry     

Changes of lipids in sardine (Sardinella gibbosa) muscle during iced storage

Changes in lipids of sardine (Sardinella gibbosa) muscle during 15 days of iced storage were investigated. Lipid deterioration, lipolysis and lipid oxidation, occurred throughout the storage. The progressive peroxide formation was monitored by the increase in the absorbance band at 3600–3200 cm⁻¹ in Fourier transform infrared (FTIR) spectra and increased peroxide values were observed in sardine muscle up to 6 days of iced storage, followed by a continuous decrease from then for 9 days (P < 0.05). The increase in thiobarbituric acid reactive substances (TBARS) was noticeable throughout the iced storage (P < 0.05). However, no difference in conjugated diene (CD) of sardine muscle was found within the first 12 days of iced storage (P > 0.05). Marked decreases in unsaturated fatty acids, especially eicosapentaenoic acid (EPA; C20:5(n − 3)) and docosahexaenoic acid (DHA; C22:6(n − 3)) were observed as the storage time increased. Those changes indicated that lipid oxidation occurred in sardine muscle. A gradual increase in free fatty acid formation, with decreases in triglyceride and phospholipid contents, was found during iced storage (P < 0.05), suggesting hydrolysis induced by lipases and phospholipases.     

Retardation of haemoglobin-mediated lipid oxidation of Asian sea bass muscle by tannic acid during iced storage

Lipid oxidation mediated by haemoglobin from tilapia was monitored in washed Asian sea bass mince with and without added tannic acid (200 and 400 ppm) during 10 days of iced storage. Control samples (without tannic acid) had the highest peroxide value (PV) within the first 2 days and possessed the greater amount of thiobarbituric acid-reactive substances (TBARS) throughout the storage of 10 days (p < 0.05). With addition of tannic acid, the lipid oxidation of washed Asian sea bass mince was retarded, especially when the higher level (400 ppm) was used, as evidenced by lowered PV and TBARS. The retarded formation of volatile lipid oxidation products in the samples with added 400 ppm tannic acid was found. Sensory analysis revealed that samples with added 400 ppm tannic acid possessed lower fishy odour score, compared with the control sample and that with added 200 ppm tannic acid (p < 0.05).     

Comparative studies on molecular changes and pro-oxidative activity of haemoglobin from different fish species as influenced by pH

Oxygenation, autoxidation as well as pro-oxidative activity of haemoglobins from tropical fish (Asian seabass, tilapia and grouper) as influenced by different pH (6, 6.5 and 7) were comparatively studied. Relative oxygenation of all haemoglobins decreased in the acidic conditions. Haemoglobin from Asian seabass was more oxygenated and stable against autoxidation at both pH 6 and 7, compared to those from tilapia and grouper. Haemoglobin from tilapia and grouper was fully oxidised at pH 6 after 120 h. Lipid oxidation of washed Asian seabass mince added with haemoglobin from various fish at different pH (6, 6.5 and 7) was monitored during 10 days of iced storage. Haemoglobins accelerated lipid oxidation more effectively at pH 6, compared with pH 6.5 and 7 as indicated by the higher peroxide value (PV) and thiobarbituric acid-reactive substances (TBARS). At the same pH values, haemoglobins from tilapia and grouper were more pro-oxidative than that from Asian seabass as evidenced by the higher PV and TBARS (P < 0.05). Volatile lipid oxidation compounds detected by gas chromatography–mass spectrometry (GC–MS) were also formed at higher rate in the washed mince added with haemoglobin from tilapia or grouper with coincidental stronger fishy odour development, compared to the control and that added with haemoglobin from Asian seabass. Thus, lipid oxidation in fish muscle was more likely governed by haemoglobin, whose pro-oxidative activity varied depending upon the pH as well as molecular properties of haemoglobin.     

Oxidation of lipid and protein in horse mackerel (Trachurus trachurus) mince and washed minces during processing and storage

Protein and lipid oxidation was followed during processing and storage of mince and washed minces prepared from horse mackerel (Trachurus trachurus). Briefly horse mackerel mince (M0) was washed with three volumes of water, mimicking the surimi production and different washed products were obtained: M1, M2 and M3, with one, two and three washing steps, respectively. The different products were characterised (i.e. lipid content, protein, water, iron, fatty acid profile and tocopherol content) and analysed for protein and lipid oxidation in order to investigate the impact of the washing steps on oxidation. Subsequently the different products were stored for up to 96 h at 5 °C and samples were taken out regularly for analysis. Lipid oxidation was investigated by measuring primary oxidation products (lipid hydroperoxides) and secondary oxidation products (volatiles). Protein oxidation was followed by determination of protein solubility, protein thiol groups and protein carbonyl groups using colorimetric methods as well as western blotting for protein carbonyl groups. Lipid and protein oxidation markers indicated that both lipid and protein oxidation took place during processing and the ranking for oxidation was as follows M0 < M1 < M2 ? M3 with M0 being significantly less oxidised than M3. Results indicated that washing creates an imbalance in the initial prooxidant-antioxidant equilibrium in the muscle tissue and contributes to the observed differences in the oxidative status of the four products obtained. In contrast, during storage of different products, lipid oxidation development was faster in M0 and the ranking was as follows M0 > M1 > M2 ? M3. Lipid and protein oxidation developed simultaneously in different minces during storage, but it was not possible to determine at which level these two reactions were coupled.     

Activity of caffeic acid in different fish lipid matrices: A review

Caffeic acid, a hydroxycinnamic acid common in different vegetable sources, has been employed as a natural antioxidant for inhibiting oxidation of fish lipids present in different food matrices. The aim of this review is to discuss the mechanisms involved in the antioxidative and prooxidative effects of caffeic acid found in different model systems containing fish lipids. These model systems include bulk fish oils, liposomes from cod roe phospholipids, fish oil emulsions, washed cod mince, regular horse mackerel mince and a fish oil fortified fitness bar. The data reported show that the antioxidant activity depends on the physical state of the lipids and the composition of the intrinsic matrix in which they are situated. Caffeic acid significantly prevented rancidity in both unwashed and washed fish mince, the latter which was fortified with haemoglobin. In the unwashed mince, the activity was however clearly dependent on the lipid to antioxidant ratio. In these systems, an important redox cycle between caffeic acid and the endogenous reducing agents ascorbic acid and tocopherol were further thought to play an important role for the protective effects. The effect of caffeic acid was also highly dependent on the storage temperature, showing higher effectiveness above than below 0 °C. Caffeic acid was not able to inhibit oxidation of bulk fish oils, fish oil in water emulsions and the fish-oil enriched fitness bar. In the liposome system, caffeic acid inhibited haemoglobin (Hb)-promoted oxidation but strongly mediated Fe²⁺ mediated oxidation. In conclusion, caffeic acid can significantly prevent Hb-mediated oxidation in fish muscle foods but its activity in food emulsions and liposomes is highly dependent on the pH, the emulsifier used and the prooxidants present.     

Effect of myoglobin from Eastern little tuna muscle on lipid oxidation of washed Asian seabass mince at different pH conditions

The effect of pH (6.0, 6.5, and 7.0) on lipid oxidation in washed Asian seabass (Lates calcarifer) mince mediated by oxymyoglobin from the dark muscle of little Eastern tuna (Euthynnus affinis) during 8 d of refrigerated storage was studied. Metmyoglobin formation and discoloration increased with increasing storage time and the changes were more pronounced at lower pH. The highest lipid oxidation and off-odor development were observed when myoglobin was incorporated in washed mince at pH 6.0. At low pH, oxidation of myoglobin took place and lipid oxidation in washed mince was enhanced. This was concomitant with the increased fishy and rancid off odor in the sample containing myoglobin, especially at pH 6.0. Washed mince containing myoglobin at pH 6.0 had 1-octen-3-ol and hexanal as the major volatile compounds. Thus, postmortem pH and myoglobin played an essential role in lipid oxidation and off odor in fish muscle during the extended storage. PRACTICAL APPLICATION: Myoglobin plays a role in the color of fish muscle. The change of myoglobin affects not only consumer acceptability, but also lipid oxidation as well as odor. The control of pH of muscle could be a potential means to lower the lipid oxidation mediated by myoglobin. As a consequence, the prime quality of fish with a negligible fishy odor could be maintained during postharvest handling or storage.     

Suitability of saturated aldehydes as lipid oxidation markers in washed turkey meat

The aim of this study was to evaluate the suitability of saturated aldehydes as lipid oxidation markers in washed turkey muscle, by means of headspace solid phase microextraction-gas chromatography (HS-SPME-GC); the results were compared with the widely used thiobarbituric acid-reactive substances (TBARs) method. Changes in TBARs, propanal and hexanal concentrations were determined over time in a model system consisting of turkey muscle washed with a sodium phosphate buffer (pH 5.6). To stop oxidation from occurring during analysis, an antioxidant mixture (EDTA, trolox and propyl gallate) was added immediately before analyses. After antioxidant addition, propanal and TBARs concentrations did not increase during 8 h of further storage, while an unexpected decrease in hexanal was observed. To determine if aldehydes were interacting with washed turkey muscle, hexanal and propanal were added to either phosphate buffer or washed muscle and concentrations were monitored for 24 h. Neither propanal nor hexanal decreased in the phosphate buffer over time, but the headspace concentration of propanal and hexanal in washed turkey muscle were markedly lower (76% and 96%, respectively) at time zero and continued to decreased up to 24 h of storage. Because of this decrease in headspace aldehyde concentrations, TBARs were found to be a more sensitive and accurate marker of oxidative deterioration in washed turkey muscle.     

Lipid oxidation and fishy odour development in protein hydrolysate from Nile tilapia (Oreochromis niloticus) muscle as affected by freshness and antioxidants

Lipid oxidation and fishy odour development in protein hydrolysate from fresh and ice-stored Nile tilapia (Oreochromis niloticus) were investigated. During iced storage of 18 days, heme iron content decreased with a concomitant increase in non-heme iron content (P < 0.05). Peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) values increased. Phospholipid content decreased with a corresponding increase in free fatty acid content. The results suggested that lipid hydrolysis and oxidation took place during storage. When protein hydrolysates were produced from fresh and 18 days ice-stored Nile tilapia muscle, higher lipid oxidation and fishy odour/flavour along with higher amount volatile compounds were obtained in hydrolysate for unfresh sample (P < 0.05). However, the addition of mixed antioxidants during hydrolysis process markedly lowered lipid oxidation, b^∗, ΔC^∗, ΔE^∗ values, fishy odour/flavour as well as the formation of volatile compounds in the resulting hydrolysates prepared from both fresh and unfresh samples. Therefore, hydrolysate from Nile tilapia muscle with reduced fishy odour and lighter colour could be prepared by using fresh fish and incorporation of mixed antioxidants during hydrolysis.     

Changes in heme proteins and lipids associated with off-odour of seabass (Lates calcarifer) and red tilapia (Oreochromis mossambicus × O. niloticus) during iced storage

Changes in heme proteins and lipids associated with off-odour development in seabass (Lates calcarifer) and red tilapia (Oreochromis mossambicus × O. niloticus) muscles during 15 days of iced storage were studied. Fresh seabass contained the higher contents of myoglobin and heme iron, compared with red tilapia (P < 0.05). An increase in metmyoglobin proportion was observed during storage. After 3 days of storage, a decreased heme iron content and a concomitant increase in non-heme iron content were noticeable in both fish (p < 0.05). Oxidation of myoglobin and released non-heme iron were associated with lipid oxidation. The increases in oxidation products and free fatty acids were observed as the storage time progressed. Fishy and rancid odours were detected at day 6 of storage for both fish and a higher intensity was found in seabass muscle. Thus, the off-odour in fish muscle was mostly governed by lipid oxidation and species specific.     

Changes in lipids and fishy odour development in skin from Nile tilapia (Oreochromis niloticus) stored in ice

Changes in lipids, lipoxygenase activity and fishy odour development in the skin of Nile tilapia (Oreochromis niloticus) during iced storage of 18 days were monitored. Triacylglycerol content of skin decreased with coincidental increases in free fatty acid, monoacylglycerol, diacylglycerol and phospholipid contents during storage (p < 0.05). During iced storage, peroxide value increased at day 9 and subsequently decreased up to 18 days (p < 0.05). Thiobarbituric acid reactive substances values and lipoxygenase activity increased throughout 18 days of iced storage (p < 0.05). With increasing storage time, a progressive formation of hydroperoxide was found as evidenced by the increase in amplitude of peak at 3600–3200 cm⁻¹ in Fourier transform infrared spectra. Those changes indicated that lipid oxidation took place during iced storage. The increase in fishy odour of skin was observed as the storage time increased. The development of fishy odour in Nile tilapia skin during iced storage was mostly governed by lipid oxidation via autoxidation or induced by lipoxygenase. Thus, the extended storage time of whole fish resulted in the pronounced changes in lipids and the increased fishy odour in the skin.     

Inhibition of haemoglobin-mediated lipid oxidation in washed cod muscle and cod protein isolates by Fucus vesiculosus extract and fractions

The effects of Fucus vesiculosus extract and fractions towards haemoglobin- (Hb-) catalysed lipid oxidation in washed cod muscle system and cod protein isolates during ice storage were examined. The extract and fractions were characterised in terms of total phlorotannin content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion-chelating ability and reducing power. Progression of oxidation was followed by determining rancid odour, thiobarbituric acid reactive substances (TBARS), redness and volatile oxidation compounds by gas chromatography (GC). In both washed cod muscle and protein isolates, phlorotannin-enriched ethyl acetate (EtOAc) fraction showed higher inhibitory effect than crude 80% ethanol (EtOH) extract. The addition of oligomeric phlorotannin-rich subfraction (LH-2) separated by Sephadex LH-20 chromatography, completely inhibited the initiation of lipid peroxidation in both systems throughout the entire study period (8 days). Its effectiveness at 300 mg/kg level was comparable to that of 100 mg/kg propyl gallate (PG), a highly effective synthetic antioxidant in muscle foods. Although polymeric phlorotannin-rich subfraction (LH-5) had similar level of TPC and chemical antioxidant activities as oligomeric subfraction LH-2, it was far less efficient in model systems. These results suggest that other factors rather than the intrinsic reactivity toward radicals could be responsible for the inhibitory effect of phlorotannins on lipid oxidation in fish muscle. This study highlights the great potential of oligomeric phlorotannins as novel natural antioxidants in fish and fish products.     

Assessing Low Redox Stability of Myoglobin Relative to Rapid Hemin Loss from Hemoglobin

Bovine myoglobin (Mb) auto‐oxidized 11‐fold faster at pH 5.7 compared to bovine hemoglobin (Hb). Replacement of Ser(F7) in bovine Mb with positively charged or large apolar residues decreased auto‐oxidation rates (2‐ to 4‐fold) in comparison with wild‐type Mb (P < 0.05). However, the same substitutions increased hemin loss rate (15‐ to 28‐fold), indicating that hydrogen bonding between Ser(F7) and the heme‐7‐propionate is critical for stabilizing protoporphyrin in the globin. The anchoring of Ser(F7) to the heme‐7‐propionate in the proximal pocket of Mb is suggested to expose the distal pocket to solvent molecules that accelerate auto‐oxidation. The rate of hemin loss from metHb at pH 5.7 was 68‐fold faster compared to metMb. The ability of Ser(F7) and His(FG3) in Mb to form stabilizing contacts with the heme‐7‐propionate maintains hemin within the globin whereas Leu(F7) and Leu(FG3) of Hb cannot form stabilizing contacts which results in low hemin affinity. MetHb promoted lipid oxidation more effectively in washed muscle at pH 5.7 compared to metMb (P < 0.05). The greater ability of bovine metHb to promote lipid oxidation is likely due to its enhanced rate of hemin dissociation compared to bovine metMb.     

Lipid and colour stability of M. longissimus muscle from lambs fed camelina or linseed as oil or seeds

Colour and lipid stability of M. longissimus dorsi (LD) from sheep fed diets containing different lipid sources (Megalac (MG), camelina oil (CO), linseed oil (LO), NaOH-treated camelina seed (CS), NaOH-treated linseed (LS) or CO treated with ethanolamine (CA)) were examined. After 100 days on-feed, samples of LD were collected, fatty acid profile determined and colour and lipid oxidation (2-thiobarbituric acid reactive substances; TBARS) measured during retail display in high oxygen packaging. The LS ration was most effective in increasing the 18:3n − 3 and conjugated linoleic acid (CLA) concentration in muscle. Within camelina, CA resulted in the highest 18:3n − 3 and lowest CLA concentration in muscle. There was no difference in colour stability. Oil (seed) supplementation increased TBARS compared to MG in the early part of display while linseed-based rations tended to cause higher TBARS than camelina-based rations. Higher muscle 18:3n − 3 concentration was associated with higher oxidation during early retail display but this was not reflected in a loss of colour stability.     

Effects of hemopexin on hemin and hemoglobin-mediated lipid oxidation in washed fish muscle

Hemopexin (Hx) was isolated from pig blood using hemin-agarose chromatography. The effect of addition of Hx on hemin and hemoglobin (Hb) mediated lipid oxidation in washed cod muscle was investigated during iced storage at pH 5.5. At a 1:1 ratio of Hx to hemin, lipid oxidation as measured by development of thiobarbituric acid reactive substances (TBARS) was not significantly inhibited (p = 0.12). This may be attributed to a lack of hemin binding by Hx due to influence of pH and hemin precipitation. At a 1:2 ratio of Hx to Hb on a heme basis, TBARS development was significantly inhibited (p < 0.01) but not prevented. Equimolar addition of bovine serum albumin (BSA) in place of Hx did not inhibit TBARS development (p = 0.43). Hemin release from porphyrin-containing Hx, i.e. holoHx, and ferric sperm whale myoglobin (Mb) was measured at 37 °C, pH 5.7. Hemin release rates of 0.27 h^?1 and 0.05 h^?1, were calculated for holoHx and Mb, respectively. While the hemin affinity of Hx was greater than published values for trout Hb, the relatively low value measured for Hx compared to Mb may be caused by a combination of low pH and the absence of NaCl in the assays.     

Effect of grape antioxidant dietary fibre on the prevention of lipid oxidation in minced fish: Evaluation by different methodologies

The effect of grape antioxidant dietary fibre (GADF) addition to minced fish muscle (MFM) on lipid stability during frozen storage (6 months) was studied. Concentrations of 0%, 2%, and 4% GADF were added to MFM samples. Analyses were carried out immediately after preparation of samples and during and after storage at −20 °C. GADF was characterized in terms of dietary fibre, total polyphenols and antioxidant capacity, and multifunctional antioxidant assays were carried out on all the MFM samples. The addition of red grape fibre considerably delayed lipid oxidation in minced horse mackerel muscle during the first 3 months of frozen storage.     

Effects of high pressure/thermal treatment on lipid oxidation in beef and chicken muscle

Lipid oxidation was studied in beef and chicken muscle after high pressure treatment (0.1–800 MPa) at different temperatures (20–70 °C) for 20 min, prior to storage at 4 °C for 7 days. Pressure treatment of beef samples at room temperature led to increases in TBARS values after 7 days storage at 4 °C; however, the increases were more marked after treatment at pressures ?400 MPa (at least fivefold) than after treatment at lower pressures (less than threefold). Similar results were found in those samples treated at 40 °C, but at 60 °C and 70 °C pressure had little additional effect on the oxidative stability of the muscle. Pressure treatments of 600 MPa and 800 MPa, at all temperatures, induced increased rates of lipid oxidation in chicken muscle, but, in general, chicken muscle was more stable than beef to pressure, and the catalytic effect of pressure was still seen at the higher temperatures of 50 °C, 60 °C and 70 °C. The addition of 1% Na₂EDTA decreased TBARS values of the beef muscle during storage and inhibited the increased rates of lipid oxidation induced by pressure. The inhibition by vitamin E (0.05% w/w) and BHT (0.02% w/w), either alone or in combination, were less marked than seen with Na₂EDTA, suggesting that transition metal ions released from insoluble complexes are of major importance in catalysing lipid oxidation in pressure-treated muscle foods.