Electron spin resonance spectroscopy for determination of the oxidative stability of food lipids

Evaluation of the oxidative stability of food lipids based on the tendency of formation of radicals is shown to be possible using electron spin resonance (ESR) spectroscopy and the spin-trapping technique. Induction time can be determined for mildly accelerated conditions (50°C for lipid fraction from mayonnaise enriched with fish oil), and the length of the induction time decreases during storage and γ-tocopherol depletion. The protection by ethylenediaminetetraacetic acid against initiation of lipid oxidation is also detected in the new assay. For more oxidatively stable lipids (butter, rapeseed oil, dairy spread) the mildly accelerated conditions can be used in the assay, provided that difference in signal height for fixed times replaces determination of induction time. ESR spin trapping provides a sensitive method for evaluating the oxidative stability of food lipids. Detection of radicals in the lipid as an early event in oxidation allows mild conditions to be used, and future experiments should also include sensory evaluation in relation to determination of practical shelf life.     

Formation of Polar Compounds During Deep‐frying—Determination by 1H NMR and ESR

The present study aims to elucidate the factors influencing the generation of polar compounds in oils during deep‐frying. Oils with different fatty acid compositions, including palm oil (PO), refined palm kernel oil (RPKO), and refined coconut oil (RCO), are applied in successive frying processes. ¹H NMR spectra reveal that heated PO has a higher percentage of allyl acyl group and is more prone to formation of non‐polar dimeric triglycerides as compared to other types of oils. In addition, electron spin resonance (ESR) spectra indicate that alkyl radicals are more predominant than alkoxy radicals in heated PO. In contrast, RPKO and RCO are inclined to generation of alkoxy radicals during the thermal treatment. The results reveal that oils with high unsaturated fatty acid content are more prone to generation and oxidation of non‐polar dimeric triglycerides. Practical Applications: The change in free radical profile and concentration is one of the indicators of lipid oxidation and polymerization. Alterations in the levels of alkyl and alkoxyl radicals, revealed by ESR, can be used to illustrate the formation of polar compounds in deep‐fried oils with different fatty acid compositions. The percentage of allyl acyl group, revealed by ¹H NMR, can be used to predict the generation of polar compounds. Therefore, this study provides useful information for the development of different methods to reduce polar compound formation in oils during thermal processing depending on the fatty acid composition of different deep‐fried oils.     

Kinetic Study of Hydroperoxide Degradation in Edible Oils Using Electron Spin Resonance Spectroscopy

Lipid oxidation is a complex phenomenon involving free radicals which are highly reactive molecular species. The life-time of these radical species is extremely short and their detection is therefore difficult. Several electron spin resonance (ESR) spectroscopy methodologies make it possible to identify, quantify and measure the reactivity of radical species formed during oxidation–reduction reactions. In this study we took advantage of the specificity of ESR spectroscopy to detect radical compounds in order to determine the rate constants of hydroperoxide degradation, a key reaction involved in lipid oxidation. The interaction of 5-doxyl stearic acid and lipid-derived radicals was studied by following the intensity of ESR spectra. A kinetic model was developed to simulate data analysis obtained by ESR and values of rate constants for hydroperoxide degradation were determined at 100 and 110 °C. This quantitative approach of ESR spectroscopy has produced useful information about new rate estimates for hydroperoxide degradation in edible oils.     

Effectiveness of antioxidants in preventing oxidation of palm oil enriched with heme iron: A model for iron fortification in baked products

Bakery products such as biscuits, cookies, and pastries represent a good medium for iron fortification in food products, since they are consumed by a large proportion of the population at risk of developing iron deficiency anemia, mainly children. The drawback, however, is that iron fortification can promote oxidation. To assess the extent of this, palm oil added with heme iron and different antioxidants was used as a model for evaluating the oxidative stability of some bakery products, such as baked goods containing chocolate. The palm oil samples were heated at 220°C for 10 min to mimic the conditions found during a typical baking processing. The selected antioxidants were a free radical scavenger (tocopherol extract (TE), 0 and 500 mg/kg), an oxygen scavenger (ascorbyl palmitate (AP), 0 and 500 mg/kg), and a chelating agent (citric acid (CA), 0 and 300 mg/kg). These antioxidants were combined using a factorial design and were compared to a control sample, which was not supplemented with antioxidants. Primary (peroxide value and lipid hydroperoxide content) and secondary oxidation parameters (p‐anisidine value, p‐AnV) were monitored over a period of 200 days in storage at room temperature. The combination of AP and CA was the most effective treatment in delaying the onset of oxidation. TE was not effective in preventing oxidation. The p‐AnV did not increase during the storage period, indicating that this oxidation marker was not suitable for monitoring oxidation in this model.     

Lipid oxidation inhibiting capacities of blackseed essential oil and rosemary extract

Lipid oxidation is the main deterioration process that occurs in vegetable oils containing lipid molecules with polyunsaturation. The aim of the present study was to investigate the possible effects of blackseed (Nigella sativa L.) essential oil (BEO) and rosemary (Rosmarinus officinalis L.) extract (RE) on stabilization of sunflower oil under accelerated storage conditions. RE was obtained by soxhlet extraction using methanol, whereas BEO was hydrodistilled from the blackseed extract obtained by extraction using petroleum ether (b.p. 40–60°C). The results indicate that both extracts stabilize sunflower oil to a certain extent, the extent being greater with RE than with BEO based on measurements of peroxide value, p‐anisidine value, amount of nonoxidized linoleic acid in saponified oil samples by HPLC‐DAD, concentration of conjugated diene hydroperoxide, and UV light absorption. The oil stabilizing effect of BEO against lipid oxidation, especially at concentrations of 0.06 and 0.1 g/100 g oil was found to increase as the temperature increased, according to principal component analysis results. Based on its strong lipid oxidation inhibiting capacity, RE can be used as a potential natural extract for stabilizing sunflower oil against oxidation. BEO, at carefully selected concentrations, can be suggested as an alternative supplement of plant origin for improving oil stabilization. Practical applications: Sunflower oil is a widely used vegetable oil for cooking and frying, and has a high linoleic acid content of about 40–70%. Here we show that sunflower oil can be stabilized by adding rosemary and blackseed extracts. The treatment can be useful especially in applications which require heating the oil to high temperatures.     

热提质褐煤预氧化后自燃特性变化及其自由基原位分析

针对不同温度热提质褐煤新鲜样及其自然环境中储存一定时间后的预氧化样，利用订制的自燃测定实验装置测得各样品的自燃倾向性差异，采用X-band电子自旋共振波谱仪进行自由基动态原位分析，研究预氧化对热提质褐煤自燃特性的影响。结果表明，热提质温度升高，新鲜样的自燃倾向性降低。但随着储存时间延长，105~500℃热提质褐煤发生预氧化，自燃倾向性不同程度地增加，400℃热提质后的预氧化样与原煤一样易自燃。在自燃氧化过程中，与新鲜样不同，预氧化样的吸氧量增加，自由基浓度也增加。预氧化不仅使热提质褐煤自由基增加；且在自燃的加速氧化阶段后期，不断产生自由基，维持氧化自热升温。而400℃热提质褐煤新鲜样在自燃氧化初期消耗大量小分子自由基，损失达80%，因不能持续产生活性自由基难以维持自燃氧化反应进行。     

氧化锌分散体光反应产生的自由基中间体

溶液中光诱导的电子转移反应已进行了大量的研究。而半导体粉末在水相或非水溶剂中的光化学研究也与自俱增。这种光化学与成像体系、太阳能转换以及光催化或污物的光降解有关。因此,越来越引起人们的重视。 Harbour和Hair用自旋捕捉技术研究了在ZnO-H_2O的分散体系中光合成过氧化氢,并检测到羟基自由基。他们认为氧化锌可能通过水的氧化将O_2还原成H_2O_2,从而使太阳能转换成化学能。人们自然会想到ZnO在其它溶剂中是否-也能观察到溶剂光氧化     

Free radical involvement in the oxidation of Victorian brown coal

Electron spin resonance spectroscopy has been used to investigate free radical involvement in the oxidation of Victorian brown coal. Fresh, wet Yallourn brown coal exhibits two superimposed e.s.r. signals due to organic free radicals. The free radicals responsible for the narrow signal interact reversibly with atmospheric oxygen while the broad signal changes intensity dramatically during prolonged exposure to air. Coal:oxygen complex formation may explain these observations. Results also suggest that the free radical content of brown coal increases as a consequence of vacuum drying.     

Volatiles and Tendency of Radical Formation of Cold‐Pressed Brazil Nut Oil During Ambient Storage

Cold‐pressed Brazil nut oil (BNO) is a value‐added nontimber forest coproduct of market interest, despite the lack of knowledge about its ambient storage stability. The objective of the present study was to investigate oxidative changes in BNO stored in clear and brown glass bottles under retail conditions with a focus on monitoring for the first time the formation of volatile aroma compounds (VAC), the depletion of tocopherol analogs, and the tendency of radical formation. VAC, whose formation continuously increased and correlated with storage time, were detected using headspace‐solid‐phase microextraction‐gas chromatography–mass spectrometry. Depletion of tocopherols was more pronounced for the oil stored in clear glass bottles and the content loss was higher for alpha‐tocopherol than for gamma‐tocopherol. Furthermore, the tendency of radical formation was detected using a spin‐trapping electron spin resonance spectroscopy method, which was demonstrated to be a simpler, faster, and less sample‐demanding tool to monitor the oxidative status of BNO stored under retail conditions and its results significantly correlated with those obtained using traditional methods for the determination of primary oxidation products. However, the simultaneous measurement of key off‐flavor volatiles is recommended for samples with unknown history.     

Stable free radical from plant litter decomposing in water

The presence of a stable radical species in both fresh straw and that which had been submerged in aerated water for up to six months has been demonstrated using electron paramagnetic (spin) resonance (EPR or ESR) spectroscopy. A radical signal was associated also with material shown to contain straw lignin markers, which was leached from the rotting straw into surrounding water. Fresh straw treated with strong alkali to remove phenolics did not show a radical signal. The possible effect of a dissolved stable free radical is discussed in relation to the antagonistic effect of rotting straw on algal and cyanobacterial growth in water to which straw has been added as a nuisance algal control agent.     

有机锡化合物光解活泼自由基的ESR研究

本文用自旋捕捉技术与ESR相结合的方法,研究了六种有机锡化合物的光解反应历程。结果表明:有机锡化合物紫外光照时,碳—锡键发生均裂,产生碳中心和锡中心自由基。其中碳中心自由基易被捕捉剂α-苯基-N-特丁基氮氧化物(PBN)或2,3,5,6-四甲基亚硝基苯(ND)所捕获;而锡中心自由基可与菲醌形成稳定的环状加合物。由于环状加合物中配体的不同,产生的空间效应也不同,从而导致加合物中的未偶电子云密度发生变化,给出相异的ESR信号。     

Electron spin resonance spectroscopy: a promising method for studying lipid oxidation in foods

Oxidative reactions that involve lipids are among the most important causes of deterioration of foods and affect both their shelf life and their organoleptic and nutritional properties. Radicals are intermediate key‐constituents in these reactions. Their detection is therefore essential in order to understand and predict early stages of these destructive oxidations. Several Electron Spin Resonance (ESR) methodologies (direct and indirect) make it possible to identify, quantify and measure the reactivity of radical species formed during oxidative‐reductive reactions. They are also used to evaluate the antiradical activity of antioxidants delaying lipid oxidation. An overview on the use of different ESR methodologies to study lipid oxidation in foods is reported in this paper.     

Free radical polymerization and lipid binding of lysozyme reacted with peroxidizing linoleic acid

Insolubilization and polymerization of proteins exposed to peroxidizing lipids may be due either to cross-linking with incorporation of fragments of the lipid oxidation products, or to free radical transfer from lipid to protein and subsequent free radical polymerization of protein. The second mechanism which has been proposed was inferred from measurements of electron spin resonance signals in proteins. In this study, uniformly labeled linoleic acid, [¹⁴C(U)] LA, was reacted with lysozyme. Volatile oxidation products of LA were also used in some experiments. Incubation was done in the absence of water. Oligomers of lysozyme, as well as the monomer, were isolated after incubation, and the [¹⁴C] label incorporated into each fraction was determined. The results show that the dominant mechanism of protein polymerization after exposure to peroxidizing linoleic acid is the transfer of free radical from lipid to protein, and subsequent free radical polymerization.     

Carbon radicals generated by solid polymers: Electron spin resonance spectroscopy for detection of species in water

Radicals generated in plastic medical devices (solid phase) by γ-rays or electron-beam irradiation during sterilization are known to cause oxidation of protein drugs, resulting in a loss or reduction in drug efficacy. The generation of radical species in water by the radical species in solid polymers has not been proved. Using electron spin resonance (ESR) spectroscopy, we confirm the generation of new radicals in water by γ-ray irradiated cyclic olefin polymers (COP). ESR measurements are obtained using 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL) as a spin probe and 5-(2,2-dimethyl-1,3-propoxycyclophosphoryl)-5-methyl-1-pyrroline N-oxide (CYPMPO) as a spin trap, in which the irradiated COP was immersed. The ESR signals indicate the TEMPOL radicals decline over time, suggesting the generation of new radicals. Conversely, the characteristic ESR signals of the adduct formed by the reaction between CYPMPO and the hydroxyl radical are observed. Thus, hydroxyl radicals are generated because of the migration of the radicals from COP to water. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48604.     

Free Radical Scavenging and Cellular Antioxidant Properties of Astaxanthin

Astaxanthin is a coloring agent which is used as a feed additive in aquaculture nutrition. Recently, potential health benefits of astaxanthin have been discussed which may be partly related to its free radical scavenging and antioxidant properties. Our electron spin resonance (ESR) and spin trapping data suggest that synthetic astaxanthin is a potent free radical scavenger in terms of diphenylpicryl-hydrazyl (DPPH) and galvinoxyl free radicals. Furthermore, astaxanthin dose-dependently quenched singlet oxygen as determined by photon counting. In addition to free radical scavenging and singlet oxygen quenching properties, astaxanthin induced the antioxidant enzyme paroxoanase-1, enhanced glutathione concentrations and prevented lipid peroxidation in cultured hepatocytes. Present results suggest that, beyond its coloring properties, synthetic astaxanthin exhibits free radical scavenging, singlet oxygen quenching, and antioxidant activities which could probably positively affect animal and human health.     

Histological localization of oil palm fruit lipase

Development of free fatty acids (FFA) and subsequent deterioration of palm oil quality can be attributed to lipase, an active hydrolytic enzyme present in the mesocarp of oil palm fruit. A histochemical method was used to localize this enzyme in the mesocarp. FFA released from endogenous glycerides were precipitated as lead salt. The deposits were located on the membranes of oil bodies. Enzymatic activity in the mesocarp was also studied under different conditions of fruit storage: ambient temperature, freezing, etc. The enzyme was active after freezing but less active under ambient conditions of fruit storage. Investigations on fruit development indicated that the enzyme activity started appearing between 16–20 weeks after anthesis (WAA) which synchronized with the beginnning of lipid formation. These histochemical studies were supported wity biochemical data on FFA.     

Thermal crosslinking behavior of poly (aryl ether ketone) copolymers containing naphthalene moieties

A series of poly[(ether ether ketone)‐co‐(ether naphthalene ether ketone)] (P(EEK‐co‐ENEK)) copolymers were heated under a variety of conditions. The thermal crosslinking behavior was monitored by differential scanning calorimetry (DSC), electron spin resonance (ESR) and wide‐angle X‐ray diffraction (WAXD). The results indicate that under a non‐oxidative environment such as nitrogen P(EEK‐co‐ENEK) is more stable, while under oxidative conditions a crosslinking reaction takes place that causes a reduction in the crystallizability of the copolymers, and an increase in the concentration of free radicals on the copolymer. ESR results suggest that the crosslinking reaction proceeds via free radicals. Subsequently two kinds of free radicals were characterized: one is an RO^? type free radical and the other is a naphthalene ring free radical. Copyright © 2004 Society of Chemical Industry     

Oxidative Stability of Pistachio (Pistacia vera L.) Paste and Spreads

In this study, pistachio (Pistacia vera L.) spread was prepared using pistachio paste, icing sugar, and red palm oil. Pistachio paste, pistachio spread without emulsifier, and pistachio spread with 1.5 % monoglycerides (Dimodan HP‐M, Danisco, Malaysia) were investigated to determine their oxidative stability by subjecting them to accelerated autoxidation conditions (60 °C). Pistachio paste and pistachio spread exhibit a great susceptibility to lipid oxidation due to their high‐fat content. However, it was found that the high oleic and palmitic contents of the oil increase its stability. In all the pistachio products examined, Totox values were higher than 10 (the maximum permitted level) after 25 days of storage. In none of the products was (E,E)‐2,4‐decadienal detected on day 0. After 25 days of storage, however, the peak area percentages rose to 0.13, 0.19, and 0.18 in pistachio paste, pistachio spread without emulsifier, and pistachio spread with emulsifier, respectively. Based on color investigations, the a value (i.e., less greenish) was increased in all the pistachio samples at the end of the storage period. Based on the results obtained from sensory evaluations as well as GC–MS, free fatty acid (FFA), and color measurements, it may be concluded that the shelf life of pistachio products is 20 days at 60 °C.     

ESR spectroscopic evaluation of void parts in microheterogeneously crosslinked monomethacrylate/dimethacrylate resins by using copper(II) tetraphenylporphyrin spin probe

As part of continuing studies concerned with the elucidation of three‐dimensional network formation mechanism in the free‐radical crosslinking polymerization of multivinyl compounds, void parts in microheterogeneously crosslinked monomethacrylate/dimethacrylate resins were evaluated by ESR spectroscopy. Thus, the usefulness of copper(II) tetraphenylporphyrin (CuTPP) as a spin probe was verified for the evaluation of void parts under specified polymerization conditions. The correlation between ESR spectra and microheterogeneity of networks formed under various conditions was examined by changing the factors influencing resultant network structures, including monomer concentration, crosslinker content and flexibility of network segments. The mobility restriction of CuTPP by network formation was reduced noticeably under the conditions introducing the inhomogeneous network as a result of the exclusion of CuTPP from densely crosslinked network parts to void ones in cured resins.     

Profiles of volatile compounds in milk containing fish oil analyzed by HS‐SPME‐GC/MS

Long‐chain polyunsaturated fatty acids (LC‐PUFA) have various positive biological effects. Fish oil represents a major source of LC‐PUFA; therefore it is extensively used to enrich food products as, for example, infant formulae, dairy products and fruit juices. However, in the presence of oxygen and metals, LC‐PUFA readily degrade, producing off‐flavors and decreasing the nutritional value of the product. The deterioration of sensory properties (taste and odor) can be easily perceived by the consumer, due to the formation of volatile compounds that are formed by decomposition of lipid hydroperoxides, also known as primary oxidation products. In this study, we used the headspace solid‐phase microextraction‐gas chromatography/mass spectrometry technique (HS‐SPME‐GC/MS) to characterize and quantify volatile compounds in a food matrix supplemented with fish oil. We demonstrated that the HS‐SPME‐GC/MS method is a valuable tool to monitor lipid oxidation at early stages. We identified t‐2‐hexenal and c‐4‐heptenal as possible oxidation markers during the storage of milk enriched with 5% of cod oil.