Effects of oxygen and antioxidants on the lipid oxidation and yellow discolouration of film from red tilapia mince

BACKGROUND: Generally, biodegradable films from fish muscle protein become yellow after preparation. This discolouration is more likely associated with lipid oxidation and can be prevented by minimising the oxidation in the films. Thus, the effects of oxygen and antioxidants on lipid oxidation and yellow discolouration of film from red tilapia mince during storage were investigated. RESULTS: Both films prepared at pH 3 and 11, and kept under atmosphere containing 100% N₂ had the lowest TBARS value with the concomitant lowest b* and ΔE* values during storage (P < 0.05), when compared with other films kept in air and a 100% O₂ atmosphere. Films prepared at pH 3 and incorporated with antioxidants (Trolox and catechin) at all levels (100, 200 and 400 mg L^?1 film‐forming solution) had the lowest TBARS value, b* and ΔE* values during storage, indicating the retardation of lipid oxidation and yellow discolouration in films. Nevertheless, films prepared at pH 11 had no difference in TBARS values, in comparison with control film, regardless of antioxidant incorporation. Coincidentally, increases in b* and ΔE* values were observed in those films. CONCLUSIONS: Lipid oxidation was the main factor inducing yellow discolouration of film exposed to oxygen and the incorporation of antioxidants in film prepared at acidic pH was able to prevent yellow discolouration of resulting film. Copyright © 2012 Society of Chemical Industry     

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.     

Physicochemical changes of tilapia (Oreochromis niloticus) muscle during salting

The effect of wet and dry saltings on the physicochemical changes of tilapia (Oreochromis niloticus) muscle was investigated. Dry salting resulted in the higher rate of salt uptake into tilapia muscle facilitating the faster decrease in A_w (p < 0.05). The pH of both dry and wet salted fish muscles tended to decrease throughout the salting time and the lower pH was found in dry salted fish (p < 0.05). The increase in the protein content in the salting medium was found during wet salted tilapia production (p < 0.05). The TCA-soluble peptide content tended to decrease with increasing the salting time in both salting methods (p < 0.05), suggesting a leaching effect of the salting medium or the exudative loss occurred in salted tilapia. Wet salting caused the greater formation of metmyoglobin in tilapia muscle when compared to dry salting at all time points (p < 0.05) and the content of metmyoglobin increased as salting time increased in both salting methods (p < 0.05). A lowered metmyoglobin with a lowered redness index of dry salted tilapia muscle was found, indicating the continuous oxidation of metmyoglobin to other hypervalent derivatives and hence the discolouration of salted tilapia. Lipid hydrolysis and oxidation of tilapia meat occurred with varying degrees in both salting methods and these changes depended on salting time. Dry salting resulted in a higher oxidation of tilapia muscle lipid as indicated by the higher PV and TBARS throughout the salting period when compared with that of wet salting (p < 0.05). In conclusion, the physicochemical changes of tilapia muscle during salting are governed by the salting method and the salting time applied.     

Tocopherol in the lipid stability of tilapia (Oreochromis niloticus) hamburgers

Presence of tocopherol is effective for fish preservation during frozen storage, inhibiting lipid degradation by oxidation. This work evaluated the antioxidant effects of α-tocopherol in diet and postmortem addition on the final quality of hamburgers produced from tilapia fillets kept frozen for zero, 30, 60, and 90 days. Chemical composition varied within the values found for tilapia fish. The increase in α-tocopherol levels reduced the values of thiobarbituric acid reactive substances (TBARS) in the samples at all time intervals. Tocopherol supplementation in diets protected the hamburgers from lipid oxidation more effectively than postmortem addition.     

Effect of bleeding on lipid oxidation and quality changes of Asian seabass (Lates calcarifer) muscle during iced storage

Lipid oxidation, microbial load and fishy odour development in the slices of bled and un-bled Asian seabass during 15 days of iced storage were comparatively investigated. Bled samples showed the lower peroxide value (PV) and thiobarbituric acid-reactive substances (TBARS) throughout the storage period (P < 0.05). Bleeding effectively lowered the total haem and non-haem iron contents in Asian seabass slices. The release of non-haem iron was pronounced in the un-bled samples during the storage. Solid phase micro-extraction coupled with gas chromatography and mass spectrometric (SPME-GCMS) analysis revealed that the bled samples stored in ice for 15 days contained the lower amount of volatile compounds. Heptanal, the major volatile compound detected in the un-bled samples, was four-fold higher than that of bled counterparts. The contents of aldehydic compounds, including hexanal, octanal, nonanal and nonenal were also higher in the former. Bled samples had the lower fishy odour, compared with the un-bled counterparts during storage (P < 0.05). The lower total viable counts (TVC) and psychrophilic bacterial counts (PBC) were observed in the bled samples, in comparison with the un-bled ones (P < 0.05). Thus, bleeding was a potential means in retarding lipid oxidation, fishy odour development, and microbial growth of Asian seabass slices during storage in ice.     

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.     

Characterization of tilapia (Oreochromis niloticus) skin gelatin extracted with alkaline and different acid pretreatments

Tilapia production is growing worldwide and to better utilize wastes from the processing industry, one important application is production of high quality fish gelatin to meet the needs of markets that are not amenable to beef or porcine gelatin. The extraction process from tilapia skin gelatin was optimized through the use of a combination of alkali (0.3 M NaOH) with different types and concentrations of acids before thermal hydrolysis. The effects of acid pretreatments on the protein yields and the physicochemical properties of tilapia gelatin were investigated. Acid concentrations (0.01–0.20 M) influenced gelatin protein recovery: 10.52%–22.40% for citric acid, 1.92%–21.55% for acetic acid, and 4.47%–24.35% for HCl. It was possible to increase gelatin yield for each of the tested acids by adjusting the acid concentration. Gelatin viscosity and the molecular weight distribution of gelatin proteins were related to the acid concentration used. Gelatin prepared using too low a concentration (e.g. 0.01 M acetic acid or HCl) or too high a concentration (e.g. >0.05 M HCl or citric acid) yielded an extract with a smaller ratio of large molecule components, such as β-chains, and exhibited lower viscosity. The film forming properties of gelatins extracted from three acid-optimized pretreatments showed no significant difference in transparency, tensile strength and elongation at break; though the gelatin film made from 0.03 M citric acid pretreated gelatin had somewhat better water barrier property than those made with HCl or acetic acid.     

The effect of natural antioxidants on haemoglobin-mediated lipid oxidation during enzymatic hydrolysis of cod protein

Heating and changes in pH often practised during fish protein hydrolysis can cause lipid oxidation. The effect of natural antioxidants towards haemoglobin-mediated lipid oxidation during enzymatic hydrolysis of cod proteins was investigated. Different variants of a washed cod model system, containing different combinations of haemoglobin and natural antioxidants (l-ascorbic acid and Fuscus vesiculosus extract), were hydrolysed using Protease P “Amano” 6 at pH 8 and 36 °C to achieve 20% degree of hydrolysis. Lipid hydroperoxides and thiobarbituric acid reactive substances (TBARS) were analysed periodically during the hydrolysis process. The in vitro antioxidant activity of the final products was investigated. Results indicate that oxidation can develop rapidly during hydrolysis and antioxidant strategies are preferable to produce good quality products. Oxidation products did not have an impact on the in vitro antioxidant activity of the hydrolysates. The natural antioxidants inhibited oxidation during hydrolysis and contributed to the antioxidant activity of the final product.     

Effect of high-pressure treatment on lipid oxidation in turkey thigh muscle during chill storage

 Formation of secondary lipid oxidation products during chill storage of vacuum-packed (99% vacuum), pressure-treated turkey thigh muscle was found to depend on working pressure (pressure range up to 500 MPa at 10°C) and to a lesser degree on pressurization time (10 and 30 min). Pressure treatment at 400 MPa and lower pressures for 30 min (and for 10 min) resulted in less formation of thiobarbituric-acid-reactive substances (TBARS) during 6 days of storage at 5°C compared to heat treatment at 100°C for 10 min, while pressure treatment at 500 MPa for 30 min gave similar development of TBARS as did the heat treatment. The formation of TBARS during storage at 5°C was found to depend exponentially on the pressure used for treatment at both 10 min and 30 min, and apparent volumes of activation are proposed as a parameter for quantification of the effects of pressure on lipid oxidation in meat during subsequent storage. Received: 18 November 1996     

The effect of metal ions on lipid oxidation,colour and physicochemical properties of cuttlefish (Sepia pharaonis) subjected to multiple freeze–thaw cycles

The effects of different metal ions at various concentrations (0, 5, 25 ppm) on lipid oxidation, discolouration and physicochemical properties of muscle protein in cuttlefish (Sepia pharaonis) subjected to multiple freeze–thaw cycles, were investigated. Lipid oxidation of all treatments increased as the freeze–thaw cycle increased. However, the rate of the TBARS increases varied, depending on concentration, type and valency of the metal ion. Fe(II) induced lipid oxidation most effectively and its prooxidative effect was in a concentration-dependent manner. Cu(I), Cu(II) and Cd(II) showed negligible effects on lipid oxidation. The increased lipid oxidation of cuttlefish with added iron was coincidental with the increase in b* values (yellowness), especially with increasing freeze–thaw cycles. Cu(I) and Cu(II) altered cuttlefish protein sulfhydryl content and the protein solubility decreased with a concomitant increase in the disulfide bond content. The oxidative changes of proteins were observed only when a concentration of metal ions of 25 ppm was used. Those changes were more intense with increasing freeze–thaw cycles. The Ca²⁺, Mg²⁺, and Mg²⁺–Ca²⁺–ATPase activities of cuttlefish natural actomyosin decreased markedly in the presence of copper, whereas the Mg²⁺–EGTA–ATPase was increased. SDS-PAGE revealed that Cu(I) and Cu(II) induced the polymerization of muscle proteins stabilised by disulfide bond formation. However, Fe(II), Fe(III) and Cd(II) exhibited no pronounced effect on the oxidation of cuttlefish muscle proteins. Therefore, copper mainly caused the oxidation of protein, while iron induced lipid oxidation and the formation of a yellow colour in cuttlefish muscle, particularly with multiple freeze–thaw cycles.     

Effect of hydroxycinnamic acids on lipid oxidation and protein changes as well as water holding capacity in frozen minced horse mackerel white muscle

The antioxidant effectiveness of different hydroxycinnamic acids for inhibiting the formation of off-flavours associated to rancidity in minced frozen horse mackerel white muscle stored at −10 and −18 °C was studied. The influence of lipid oxidation on protein aggregation, protein denaturation and water holding capacity was also determined. Caffeic acid, o-coumaric acid and ferulic acid were the antioxidants tested. The order of antioxidant effectiveness was caffeic acid > ferulic acid > o-coumaric acid accordingly with previous results obtained in chilled horse mackerel. A strong dependence of antioxidant effectiveness to the ratio lipid/antioxidant concentration was observed. There was no evidence to suggest that antioxidants reduce protein aggregation or water holding capacity changes. Protein aggregation was not accompanied by gross protein denaturation, as monitored by differential scanning calorimetry. No evidence was found for a link between extent of lipid oxidation and degree of protein aggregation or water holding capacity changes.     

The effect of myofibrillar/sarcoplasmic protein ratios on the properties of round scad muscle protein based film

The effect of the ratios of myofibrillar protein (MP) to sarcoplasmic protein (SP) from round scad (Decapterus maruadsi) muscle on the properties of the resulting films was investigated. Tensile strength (TS) of films decreased with increasing SP content (p < 0.05). Films prepared from MP/SP ratio of 10:0 (w/w) exhibited the highest TS (p < 0.05). Elongation at break (EAB) of films prepared with SP content greater than 30% had the decreased EAB (p < 0.05). Water vapor permeability (WVP) of films increased when SP content increased up to 20% and decreased with increasing SP content up to 30% (p < 0.05). Solubility of films decreased but protein solubility increased with increasing SP contents (p < 0.05). The a*-value and ΔE* of film increased with increasing SP content. Films with all MP/SP ratios exhibited the negligible transmission to the light in UV range. Therefore, it is suggested that the type and ratio of proteins in fish muscle, both SP and MP, influenced the properties of film from round scad muscle. Results suggested that the removal of sarcoplasmic protein from fish muscle by thorough washing was an effective means to improve the mechanical properties as well as color of the fish muscle protein-based film.     

Quality loss during the frozen storage of sea salmon (Pseudopercis semifasciata). Effect of rosemary (Rosmarinus officinalis L.) extract

Lipid and protein alterations during the frozen storage (−11 °C) were analyzed in minced sea salmon muscles to evaluate the effect of the application of rosemary extract (200 and 500 mg/kg). Lipid oxidation reached maximum TBA values between 3 and 4 months of storage in untreated muscles. The main polyunsaturated fatty acids affected were 22:6-ω3, 22:5-ω3 and 20:4-ω6 acids. Phospholipid hydrolysis was also detected. Rosemary extract reduced lipid oxidation for 6 months (500 mg/kg, muscle with 10.8 g/kg lipids) or 3 months (200 mg/kg, muscle with 5.3 g/kg lipids). Myofibrillar proteins showed a decrease of extractability (80%) after 2 months of storage. Myosin denaturation was evident by DSC at 3 months, while myosin and actin peaks disappeared at 6 months. A diminution of extractable polypeptides of high molecular weight was recorded by SDS-PAGE after 3 months. The available lysine content suffered a reduction starting at 3 months of storage, suggesting some interaction involving the free amino groups of lysine. Fluorescent compounds' determination did not show changes due to the interaction of lipid oxidation products and proteins, while protein alterations could not be reduced by the rosemary extract. Furthermore, the antioxidant reduced the loss of red color in the muscle.     

Grape seed extract inhibits lipid oxidation in muscle from different species during refrigerated and frozen storage and oxidation catalyzed by peroxynitrite and iron/ascorbate in a pyrogallol red model system

The antioxidant effect of grape seed extract (GSE) was determined by assessing the bleaching of pyrogallol red (PGR) by peroxynitrite or iron/ascorbate, and the formation of lipid hydroperoxides (LOOH) and thiobarbituric acid substances (TBARS) in raw or cooked ground muscle during refrigerated or frozen storage. In PGR models, GSE was more effective than gallic acid in inhibiting oxidation. The formation of LOOH and TBARS was inhibited by GSE (0.1% and 1.0%) compared to untreated controls and samples treated with sodium tripolyphosphate. Diethylenetriaminepentaacetic acid (DTPA), alone or in combination with GSE, had no effect on LOOH or TBARS, which provides clues about the possible mechanism of action of GSE. These results show that GSE at concentrations as low as 0.1% is a very effective inhibitor of primary and secondary oxidation products in various muscle systems and has potential as a natural antioxidant in raw and cooked meat systems.     

Influences of potassium ferrocyanide on lipid oxidation of salted cod (Gadus morhua) during processing, storage and rehydration

The effects of added potassium ferrocyanide (CN) in different concentrations (2.5 ppm, 7.5 ppm and 100 ppm), in salt, on lipid oxidation in cod during salting, storage and rehydration were examined in this study. An increase in CN concentration accelerated lipid oxidation of the salted cod, as observed by increases in lipid hydroperoxides (PV) and thiobarbituric acid-reactive substances (TBARS), as well as in the development of fluorescence compounds (δF_or and δF_aq). A yellow discolouration (higher b^∗ value) of salted cod was associated with higher levels of oxidation derivatives. High correlation between PV, TBARS and free fatty acid (FFA), as well as between FFA and δF_or, was found. The results of principal component analysis showed that TBARS, b^∗ value and δF_or were the strongest indicators of lipid oxidation during salting and storage.     

罗非鱼-豆粕共沉淀蛋白的提取工艺及蛋白组成分析

以罗非鱼肉和脱脂豆粕为原料,采用p H调节法制备罗非鱼-大豆共沉淀蛋白（Co-p）,探讨溶解p H、不同质量比混合的原料、溶解时间对可溶性蛋白得率的影响及沉淀p H对蛋白沉淀得率的影响。结果表明,p H调节法回收罗非鱼-豆粕共沉淀蛋白的最佳酸溶p H2.0、3.0,碱溶p H11.0、12.0,原料比1∶1,溶解时间30 min;SDS-PAGE分析显示,可溶性共沉淀蛋白条带深/浅（极端p H2.0蛋白降解）,表明可溶性蛋白含量高/低,从分子量分布范围可知,共沉淀蛋白主要由肌球蛋白重链、7 S抗原蛋白的三个亚基、肌动蛋白、肌球蛋白轻链、11 S抗原蛋白的两个亚基和小分子水溶性蛋白组成,表示可溶性共沉淀蛋白组成齐全;酸/碱可溶性共沉淀蛋白最佳沉淀p H为4.5,在此条件下,溶解、沉淀过程的蛋白得率分别为88.05%⁹4.70%。经冷冻干燥得到共沉淀蛋白粉即Co-p（1∶1）,其蛋白含量高于85%,脂肪含量在0.84%左右,灰分含量低于4.17%;可用p H调节法回收罗非鱼-豆粕共沉淀蛋白。      

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.     

Isolation and characterisation of acid-solubilised collagen from the skin of Nile tilapia (Oreochromis niloticus)

Acid-solubilised collagen (ASC) was extracted from the skin of Nile tilapia (Oreochromis niloticus) and characterisation was studied. The results indicated that the yield of ASC was 39.4% on the basis of dry weight. This ASC was rich in glycine (35.6%). The amount of imino acids, proline and hydroxyproline, in ASC was 210 residues per 1000 residues. The ultraviolet (UV) absorption spectrum of ASC showed that the distinct absorption was at 220 nm. ASC showed transition curve at maximum temperature (T_max) of 32.0 °C in 0.05 M acetic acid, about 12 °C lower than that of calf skin collagen. Maximum solubility (0.75 mg/ml) in 0.5 M acetic acid was observed at pH 3. Solubility reached the minimum at pH 7. A sharp decrease in solubility was observed in 2% (w/v) NaCl or above. Biochemical studies indicated that ASC was composed of the α1α2α3 heterotrimers.