Lipid and volatile profiles of Finnish oat batches of pure cultivars: Effect of storage on the volatile formation.

Recent data showing the compositional variation and storage behavior among different oat batches for the purpose of food remains limited. Lipids of twenty oat flour samples of pure cultivars grown in Finland during 2019 were extracted and fractionated into neutral and polar-rich lipids. Flour was stored for nine months, and profiles of volatiles and tocols were analyzed to reveal oxidative stability. The lipid content was 5.9-8.9 g per 100 g of flour [DW] and consisted of 78.7 ± 2.5 % neutral and 21.3 ± 2.5 % polar lipids. Palmitic (16 %), oleic (36 %), and linoleic (39 %) acids were the most abundant fatty acids. Neutral lipids had more oleic and less linoleic and palmitic acids than polar lipids. The fresh samples correlated with tocols, pentanal, 2-pentylfuran, 2-heptanone, nonanal, 2-butanone, and heptanal, while stored samples were associated with 3-octen-2-one, 2-octenal, hexanal, and octanal. Lipid composition and oxidative stability are essential factors for selecting oat batches for food applications.

Antioxidative Effect of Dihydrosphingosine (d18:0) and α-Tocopherol on Tridocosahexaenoin (DHA-TAG).

Sphingoid bases have shown promise as effective antioxidants in fish oils together with α-tocopherol, and the effect has been attributed to products resulting from amino-carbonyl reactions (lipation products) between the sphingoid base amine group and carbonyl compounds from lipid oxidation. In this study, the synergistic effect of dihydrosphingosine (d18:0) and α-tocopherol was studied on pure docosahexaenoic acid (DHA) triacylglycerols with an omics-type liquid- and gas-chromatographic mass spectrometric approach to verify the synergistic effect, to get a comprehensive view on the effect of d18:0 on the oxidation pattern, and to identify the lipation products. The results confirmed that d18:0 rapidly reacts further in the presence of lipid oxidation products and α-tocopherol. α-Tocopherol and d18:0 showed an improved antioxidative effect after 12 h of oxidation, indicating the formation of antioxidants through carbonyl-amine reactions. Imines formed from the carbonyls and d18:0 could be tentatively identified.

Lipid Structure Influences the Digestion and Oxidation Behavior of Docosahexaenoic and Eicosapentaenoic Acids in the Simulated Digestion System.

Omega-3 fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are essential for human health but prone to oxidation. While esterification location is known to influence the stability of omega-3 in triacylglycerols (TAGs) in oxidation trials, their oxidative behavior in the gastrointestinal tract is unknown. Synthesized ABA- and AAB-type TAGs containing DHA and EPA were submitted to static in vitro digestion for the first time. Tridocosahexaenoin and DHA as ethyl esters were similarly digested. Digesta were analyzed by gas chromatography, liquid chromatography-mass spectrometry, and nuclear magnetic resonance spectroscopy. Besides the formation of di- and monoacylglycerols, degradation of hydroperoxides was detected in ABA- and AAB-type TAGs, whereas oxygenated species increased in tridocosahexaenoin. Ethyl esters were mainly unaffected. EPA was expectedly less susceptible to oxidation prior to and during the digestion process, particularly in sn-2. These results are relevant for the production of tailored omega-3 structures to be used as supplements or ingredients.

Enzyme-assisted aqueous extraction of fish oil from Baltic herring (Clupea harengus membras) with special reference to emulsion-formation, extraction efficiency, and composition of crude oil.

Enzyme-assisted aqueous extraction (EAAE) is a green, and scalable method to produce oil and protein hydrolysates from fish. This study investigated the role of different parameters on emulsion formation, oil recovery, and the composition of crude oil during EAAE of Baltic herring (Clupea harengus membras). Fatty acid compositions, lipid classes, tocopherols, and oxidation status of the EAAE crude oils were studied. Compared to solvent-extracted oil, EAAE resulted in a lower content of phospholipids accompanied by a 57% decrease in docosahexaenoic acid. Changing fish to water ratio from 1:1 to 2:1 (w/w) with ethanol addition led to the greatest reduction (72%) of emulsion, which resulted in an increase in oil recovery by 11%. The addition of ethanol alone, or reduction of enzyme concentration from 0.4% to 0.1% also reduced emulsion-formation significantly. Overall, emulsion reduction resulted in higher content of triacylglycerols and n - 3 polyunsaturated fatty acids in the crude oil extracted.

Docosahexaenoic acid in regio- and enantiopure triacylglycerols: Oxidative stability and influence of chiral antioxidant.

Docosahexaenoic acid (DHA) is essential for health but easily oxidized. Yet the influence of DHA's exact location (sn-1, sn-2, or sn-3) in triacylglycerols on oxidative stability is currently unknown. This is the first study comparing oxidative stability of DHA in regio- and enantiopure triacylglycerols with or without RRR-α-tocopherol. Headspace solid-phase micro-extraction with gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and nuclear magnetic resonance spectroscopy were applied. DHA in sn-2 was the most stable with or without added RRR-α-tocopherol resulting in differences in hydroperoxide formation. Without antioxidant, stability of DHA in sn-1 and sn-3 was mainly similar, with slight tendency towards better stability in sn-3. With RRR-α-tocopherol higher stability in sn-1 compared to sn-3 was observed. This points to diastereomeric interactions between RRR-α-tocopherol and DHA in sn-1. These results are highly relevant for enzymatic restructuring processes of DHA-rich fish or microalgae oil concentrates aimed for food supplements or food fortification.

Baltic herring (Clupea harengus membras) protein isolate produced using the pH-shift process and its application in food models.

In this study, protein isolate was prepared from Baltic herring (Clupea harengus membras) using alkaline pH-shift process. The aim of this research was to characterize the protein isolate and to study its potential in food models. A special focus was placed on characterization of odour profile and volatile compounds contributing to the odour profile of the protein isolate using gas chromatography - olfactometry. 2,3-Pentanedione, hexanal, 4(Z)-heptenal, 2,4(E,E)-nonadienal, and three compounds tentatively identified as 1,5(E)-octadien-3-ol, 1,5(Z)-octadien-3-ol, and 1,5(Z)-octadien-3-one were the most important odour-contributing compounds in the protein isolate (Nasal Impact Factor 83-100%, intensity 2.6-3.3 on a scale 0-4). 2-Methylpropanal, 2- and 3-methylbutanal, and three unknown compounds were less intense in the protein isolate than in the raw material, which might have contributed to the lower intensity of fishiness observed for the protein isolate (2.2 vs 3.3 on a scale 0-4). Surimi-type gels prepared from the Baltic herring protein isolate had texture properties (hardness and cohesiveness) similar to those of commercial products. Due to the abundancy of dark muscle tissue in Baltic herring, the protein isolate had a significantly lower whiteness (W = 63) compared to the commercial surimi products (W = 80-83). Increasing the solubilisation or precipitation pH did not improve the whiteness, but resulted in significantly softer, less cohesive, and less chewy gels. The findings of this study indicate that alkaline-based pH-shift processing is a potential way to increase the food application of Baltic herring.

Food Fortification Using Spray-Dried Emulsions of Fish Oil Produced with Maltodextrin, Plant and Whey Proteins-Effect on Sensory Perception, Volatiles and Storage Stability.

Fortification of foods with fish oil rich in n-3 fatty acids improves the nutritional value, but creates challenges with flavor and oxidative stability, especially during storage. Pea, soy, and sunflower proteins were used in combination with whey protein or maltodextrin to encapsulate fish oil by spray-drying. The use of whey protein compared with maltodextrin as wall material improved oxidative stability of spray-dried emulsions, although the use of whey protein increased the number of observed cracks in outer shell of the particles. Non- and encapsulated oil were used in cookies and chocolates to examine flavor characteristics by generic descriptive analysis and volatile products by solid-phase microextraction with gas chromatography-mass spectrometry. A long-term storage test at room temperature was conducted to evaluate the oxidative stability of the food models. Fortification changed the texture, odor, and flavor of the food models with fishy flavor being the most impactful attribute. For both food models, use of pea protein with maltodextrin resembled attributes of control the best. Fortification and encapsulation material also affected volatile profiles of food models. Both non-encapsulated oil and whey protein formulations performed well in regard to oxidative stability for both food models. Generally, the cookie model showed more potential for fortification than the chocolate one.

Oxidative stability, oxidation pattern and α-tocopherol response of docosahexaenoic acid (DHA, 22:6n-3)-containing triacylglycerols and ethyl esters.

DHA is most often used in supplements either in its triacylglycerol or ethyl ester form. Currently, there is only little published data on the differences in the oxidative stability and α-tocopherol response between the two lipid structures, as well as on the oxidation patterns of pure DHA. This study investigated the oxidative stability, α-tocopherol response and oxidation pattern of DHA incorporated in triacylglycerols and as ethyl esters with an untargeted approach after oxidation at 50 °C in the dark. Liquid and gas chromatographic methods with mass spectrometric detection and nuclear magnetic resonance spectroscopy were applied. DHA was more stable in triacylglycerols than as ethyl esters without α-tocopherol addition. With α-tocopherol added the opposite was observed. The oxidation products formed during triacylglycerol and ethyl ester oil oxidation were mostly similar, but also some structure-related differences were detected in both volatile and non-volatile oxidation products.

Quality of Protein Isolates and Hydrolysates from Baltic Herring (Clupea harengus membras) and Roach (Rutilus rutilus) Produced by pH-Shift Processes and Enzymatic Hydrolysis.

Fractionation is a potential way to valorize under-utilized fishes, but the quality of the resulting fractions is crucial in terms of their applicability. The aim of this work was to study the quality of protein isolates and hydrolysates extracted from roach (Rutilus rutilus) and Baltic herring (Clupea harengus membras) using either pH shift or enzymatic hydrolysis. The amino acid composition of protein isolates and hydrolysates mostly complied with the nutritional requirements for adults, but protein isolates produced using pH shift showed higher essential to non-essential amino acid ratios compared with enzymatically produced hydrolysates, 0.84-0.85 vs. 0.65-0.70, respectively. Enzymatically produced protein hydrolysates had a lower total lipid content, lower proportion of phospholipids, and exhibited lower degrees of protein and lipid oxidation compared with pH-shift-produced isolates. These findings suggest enzymatic hydrolysis to be more promising from a lipid oxidation perspective while the pH-shift method ranked higher from a nutrient perspective. However, due to the different applications of protein isolates and hydrolysates produced using pH shift or enzymatic hydrolysis, respectively, the further optimization of both studied methods is recommended.

Enzyme-Assisted Extraction of Fish Oil from Whole Fish and by-Products of Baltic Herring (Clupea harengus membras).

Baltic herring (Clupea harengus membras) is one of the most abundant commercially caught fish species from the Baltic Sea. Despite the high content of fat and omega-3 fatty acids, the consumption of Baltic herring has decreased dramatically over the last four decades, mostly due to the small sizes and difficulty in processing. At the same time there is an increasing global demand for fish and fish oil rich in omega-3 fatty acids. This study aimed to investigate enzyme-assisted oil extraction as an environmentally friendly process for valorizing the underutilized fish species and by-products to high quality fish oil for human consumption. Three different commercially available proteolytic enzymes (Alcalase®, Neutrase® and Protamex®) and two treatment times (35 and 70 min) were investigated in the extraction of fish oil from whole fish and by-products from filleting of Baltic herring. The oil quality and stability were studied with peroxide- and p-anisidine value analyses, fatty acid analysis with GC-FID, and volatile compounds with HS-SPME-GC-MS. Overall, longer extraction times led to better oil yields but also increased oxidation of the oil. For whole fish, the highest oil yields were from the 70-min extractions with Neutrase and Protamex. Protamex extraction with 35 min resulted in the best fatty acid composition with the highest content of eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3) but also increased oxidation compared to treatment with other enzymes. For by-products, the highest oil yield was obtained from the 70-min extraction with Protamex without significant differences in EPA and DHA contents among the oils extracted with different enzymes. Oxidation was lowest in the oil produced with 35-min treatment using Neutrase and Protamex. This study showed the potential of using proteolytic enzymes in the extraction of crude oil from Baltic herring and its by-products. However, further research is needed to optimize enzymatic processing of Baltic herring and its by-products to improve yield and quality of crude oil.

Effect of supercritical CO2 plant extract and berry press cakes on stability and consumer acceptance of frozen Baltic herring (Clupea harengus membras) mince.

A promising way of processing Baltic herring, Clupea harengus membras, is turning the fish into boneless mince. However, Baltic herring is prone to lipid oxidation, which possesses a challenge for industrial applications. The aim of this work was to study the efficacy of press cakes from Finnish berries and a supercritical CO2 plant extract to limit lipid oxidation during frozen storage of Baltic herring mince and to determine the impact of these additions on consumer acceptance in a fish product. Peroxide value, formation of volatile oxidation products and loss of polyunsaturated fatty acids showed that the tested natural additives decreased oxidation to a greater or similar extent as conventional antioxidants during 10-month storage. While potential of berry press cakes and plant extracts as "green label antioxidants" was shown, consumer study indicated need for further research to reach both optimal antioxidative efficacy and sensory properties.

Evaluation of the composition and oxidative status of omega-3 fatty acid supplements on the Finnish market using NMR and SPME-GC-MS in comparison with conventional methods.

Previous studies disagree on the oxidative status of omega-3 supplements. The great deviation raises concerns about quality and the methods used to monitor it. This study investigated 49 omega-3 products for their fatty acid content, lipid class and oxidative status using official methods, gas and liquid chromatography with mass spectrometry and nuclear magnetic resonance spectroscopy. With minor deviations, omega-3 fatty acid content and lipid class of all products were as declared. 24% of studied products exceeded thresholds set by The Global Organization for EPA and DHA Omega-3s for peroxide and/or p-anisidine value suggesting a compromised oxidative status. However, peroxide and/or p-anisidine value were only suitable for detection of lipid oxidation in 90% or 73%, respectively, of the products. Analysis of volatile oxidation compounds can be an alternative method for p-anisidine value. Nuclear magnetic resonance spectroscopy was shown to be a rapid method for determination of oil type and lipid class.

Effect of SPME extraction conditions and humidity on the release of volatile lipid oxidation products from spray-dried emulsions.

The aim of the study was to investigate the effects of headspace solid-phase microextraction (HS-SPME) conditions and relative humidity (RH) on the release of volatile lipid oxidation products from spray-dried emulsions. Two model spray-dried sunflower oil emulsions with a Na-caseinate-maltodextrin matrix were oxidised, stabilised at five RHs and analysed by HS-SPME-GC-MS for volatiles. Increased extraction temperature raised not only the overall release of volatile compounds but also altered the volatile profile. The obtained volatile profiles were dependent on the RH. This was mainly due to the humidity response of the matrix affecting e.g. its binding ability and hydrophilicity. Cross-linking of the emulsifying protein had a minor influence on the release. Both matrix-related factors and extraction conditions should thus be taken into account in interpretation of the HS-SPME results. On the other hand, being sensitive to changes in matrix composition and structure, the HS-SPME allows studying of matrix-related changes in foods.

Interfacial protein engineering for spray-dried emulsions - part I: effects on protein distribution and physical properties.

Distribution of protein and oil in aqueous and spray-dried emulsions and the effect of protein cross-linking on emulsion properties and matrix-water interactions were investigated. Sodium caseinate and sunflower oil were used to make emulsions which were spray dried using maltodextrin as a wall material. 3% Na-caseinate concentration showed optimum emulsion and process stability as observed in CLSM images, droplet size data and in the amount of heptane-extractable oil from spray-dried emulsions. Transglutaminase cross-linking prior to emulsification slightly increased the amount of protein both on the oil droplet interface and on the particle surface as confirmed by analysis of continuous phase protein in the feed emulsion and by XPS measurements from the powder surface. DSC and water sorption measurements were used to study the physical state of the matrix. Glass transition occurred between RH 54% and 75% at room temperature and it was not affected by cross-linking.

Interfacial protein engineering for spray-dried emulsions - part II: oxidative stability.

The aim of this work was to investigate how the oxidative stability of encapsulated oil is affected by the humidity response of a Na-caseinate-maltodextrin matrix. Furthermore, the effect of modification of the interfacial Na-caseinate layer through cross-linking was studied. For this purpose, two model spray-dried emulsions containing sunflower oil, maltodextrin, and either non-cross-linked or cross-linked Na-caseinate were stored at different relative humidities (RHs; ∼0%, 11%, 33%, 54%, and 75%). Increasing RH improved the oxidative stability of the spray-dried emulsions. This behaviour was mainly linked to the loss of individual powder particles upon caking and collapsing of the matrix at RH 75%. Oxidation of non-encapsulated surface lipids with a proportion of ca. 5% of total lipids was only twofold compared to total lipids. Excess protein on particle surfaces may have delayed oxidation, e.g., by its radical scavenging activity. Under several storage conditions, cross-linking of the protein slightly improved the oxidative stability.