Content and Bioaccessibility of Minerals and Proteins in Fish-Bone Containing Side-Streams from Seafood Industries.

With the aim to upcycle fish side-streams, enzymatic hydrolysis is often applied to produce protein hydrolysates with bioactive properties or just as a protein source for food and feed. However, the production of hydrolysates generates a side-stream. For underutilized fish and fish backbone this side-stream will contain fish bones and make it rich in minerals. The aim of this study was to assess the relative bioaccessibility (using the standardized in vitro model INFOGEST 2.0) of minerals in a dietary supplement compared to bone powder generated after enzymatic hydrolysis of three different fish side-streams: undersized whole hake, cod and salmon backbones consisting of insoluble protein and bones. Differences in the bioaccessibility of protein between the powders were also investigated. The enzyme hydrolysis was carried out using different enzymes and hydrolysis conditions for the different fish side-streams. The content and bioaccessibility of protein and the minerals phosphorus (P), calcium (Ca), potassium (K) and magnesium (Mg) were measured to evaluate the potential of the powder as an ingredient in, e.g., dietary supplements. The bone powders contained bioaccessible proteins and minerals. Thus, new side-streams generated from enzymatic hydrolysis can have possible applications in the food sector due to bioaccessible proteins and minerals.

Effect of Post-Extraction Ultrasonication on Compositional Features and Antioxidant Activities of Enzymatic/Alkaline Extracts of Palmaria palmata.

Palmaria palmata is a viable source of nutrients with bioactive properties. The present study determined the potential role of post-extraction ultrasonication on some compositional features and antioxidant properties of enzymatic/alkaline extracts of P. palmata (EAEP). No significant difference was detected in terms of protein content and recovery, as well as the amino acid composition of the extracts. The nitrogen-to-protein conversion factor of 5 was found to be too high for the seaweed and EAEP. The extracts sonicated by bath for 10 min and not sonicated showed the highest and lowest total phenolic contents (p < 0.05), respectively. The highest radical scavenging and lowest metal-chelating activities were observed for the non-sonicated sample, as evidenced by IC50 values. The extract sonicated by bath for 10 min showed the most favorable in vitro antioxidant properties since its radical scavenging was not significantly different from that of the not-sonicated sample (p > 0.05). In contrast, its metal-chelating activity was significantly higher (p < 0.05). To conclude, post-extraction ultrasonication by an ultrasonic bath for 10 min is recommended to increase phenolic content and improve the antioxidant properties of EAEP.

Screening for Metal-Chelating Activity in Potato Protein Hydrolysates Using Surface Plasmon Resonance and Peptidomics.

Metal-catalyzed lipid oxidation is a major factor in food waste, as it reduces shelf life. Addressing this issue, our study investigates the potential of hydrolysates derived from potato protein, a by-product of potato starch production, as metal-chelating antioxidants. Through sequential enzymatic hydrolysis using alcalase or trypsin combined with Flavourzyme, we produced various hydrolysates, which were then fractionated using ultrafiltration. Using a combination of peptidomics and bioinformatics, we predicted the presence of metal-chelating and free radical-scavenging peptides across all hydrolysate fractions, with a trend indicating a higher content of antioxidant peptides in lower molecular weight fractions. To validate these predictions, we utilized surface plasmon resonance (SPR) and a 9-day emulsion storage experiment. While SPR demonstrated potential in identifying antioxidant activity, it faced challenges in differentiating between hydrolysate fractions due to significant standard errors. In the storage experiment, all hydrolysates showed lipid oxidation inhibition, though not as effectively as ethylenediaminetetraacetic acid (EDTA). Remarkably, one fraction (AF13) was not significantly different (p < 0.05) from EDTA in suppressing hexanal formation. These results highlight SPR and peptidomics/bioinformatics as promising yet limited methods for antioxidant screening. Importantly, this study reveals the potential of potato protein hydrolysates as antioxidants in food products, warranting further research.

Recovery of Nutrients from Cod Processing Waters.

Liquid side-streams from food industries can be processed and used in food applications and contribute to reduce the environmental footprint of industries. The goal of this study was to evaluate the effectiveness and applicability of protein and phosphorus separation processes, namely microfiltration, ultrafiltration and flocculation, using protein-rich process waters with low (LS) and high (HS) salt content from the processing of salted cod (Gadus morhua). The application of different flocculants (chitosan lactate and Levasil RD442) were evaluated at different concentrations and maturation periods (0, 1 or 3 h). The results showed that different flocculation treatments resulted in different recoveries of the nutrients from LS and HS. Proteins in LS could be most efficiently recovered by using Levasil RD442 0.25% and no maturation period (51.4%), while phosphorus was most efficiently recovered when using Levasil RD442 1.23% and a maturation period of 1 h (34.7%). For HS, most of its protein was recovered using Levasil RD442 1.23% and a maturation period of 1 h (51.8%), while phosphorus was recovered the most using Levasil 1.23% and no maturation period (47.1%). The salt contents allowed interactions through intermolecular forces with Levasil RD442. The ultrafiltration method was effective on HS since it recovered higher percentages of nutrients in the retentate phase (57% of the protein and 46% of the phosphorus) compared to LS.

Oxidative Stability of Side-Streams from Cod Filleting-Effect of Antioxidant Dipping and Low-Temperature Storage.

Currently, side-streams (e.g., head, backbone, tail, and intestines) generated in the fish processing industry often end up as low-value products for feed applications or even as waste. In order to upcycle such side-streams, they need to be preserved to avoid oxidative degradation of the lipids between the generation point and the valorization plant. In the cod filleting industry, three main solid side-streams: viscera, heads, and backbones, are obtained. Hence, this study aimed to identify the most efficient antioxidant for preserving the cod side-streams using a dipping-based strategy prior to pre-valorization storage at low temperatures (ice and frozen storage). The dipping solutions evaluated contained: (i) a lipophilic rosemary extract (0.05% and 0.2% in 0.9% NaCl), (ii) Duralox MANC (a mixture of rosemary extract, ascorbic acid, tocopherols, and citric acid; 2% in 0.9% NaCl), and (iii) NaCl (0.9%) w/w solution. One group was not dipped. No dipping and dipping in NaCl were included as controls. The results showed a positive effect of dipping with solutions containing antioxidants as measured by peroxide value (PV), TBA-reactive substances (TBARS), and sensory profiling, e.g., rancid odor. Moreover, the oxidative stability increased with decreased storage temperature. The cod side-streams were in general most efficiently preserved by Duralox MANC, followed by the lipophilic rosemary extract (0.2%), compared to no dipping and dipping in NaCl solution and the lower concentration of the lipophilic rosemary extract (0.05%). The efficiency of the antioxidant treatments was independent of the side-stream fraction and storage temperature. Thus, using antioxidant dipping combined with low temperature storage is an efficient preservation method for maintaining the quality of the lipids in cod solid side-streams during their pre-valorization storage.

Oxidation and oxidative stability in emulsions.

Emulsions are implemented in the fabrication of a wide array of foods and therefore are of great importance in food science. However, the application of emulsions in food production is restricted by two main obstacles, that is, physical and oxidative stability. The former has been comprehensively reviewed somewhere else, but our literature review indicated that there is a prominent ground for reviewing the latter across all kinds of emulsions. Therefore, the present study was formulated in order to review oxidation and oxidative stability in emulsions. In doing so, different measures to render oxidative stability to emulsions are reviewed after introducing lipid oxidation reactions and methods to measure lipid oxidation. These strategies are scrutinized in four main categories, namely storage conditions, emulsifiers, optimization of production methods, and antioxidants. Afterward, oxidation in all types of emulsions, including conventional ones (oil-in-water and water-in-oil) and uncommon emulsions in food production (oil-in-oil), is reviewed. Furthermore, the oxidation and oxidative stability of multiple emulsions, nanoemulsions, and Pickering emulsions are taken into account. Finally, oxidative processes across different parent and food emulsions were explained taking a comparative approach.

Liquid Side Streams from Mussel and Herring Processing as Sources of Potential Income.

The seafood industry generates significant amounts of process waters which can generate value upon recovery of their nutrients. Process waters from the herring marination chain and cooking of mussels were here characterized in terms of crude composition, volatile compounds, and nutritional and potentially toxic elements. Protein and total fatty acid contents of herring refrigerated sea water (RSW) reached 3 and 0.14 g/L, respectively, while herring presalting brine (13%) reached 16.3 g/L protein and 0.77 g/L total fatty acid. Among three herring marination brines vinegar brine (VMB), spice brine (SPB), and salt brine (SMB), SPB reached the highest protein (39 g/L) and fatty acids (3.0 g/L), whereas SMB and VMB at the most had 14 and 21 g protein/L, respectively, and 0.6 and 9.9 g fatty acids/L, respectively. Essential amino acid (EAA) in marination brines accounted for up to 59% of total amino acid (TAA). From mussel processing, cooking juice had more protein (14-23 g/L) than the rest of the process waters, and in all water types, EAA reached up to 42% of TAA. For all process waters, the most abundant nutritional elements were Na, K, P, Ca, and Se. The content of all potentially toxic elements was mostly below LOD, except for As which ranged from 0.07 to 1.07 mg/kg among all tested waters. Our findings shed light on liquid seafood side streams as untapped resources of nutrients which can be valorized into food/feed products.

Starfish (Asterias rubens) as a New Source of Marine Lipids: Effect of Season, Size and Oil Extraction Methods.

The increasing demand for oils that contain health-beneficial omega-3 fatty acids calls for new resources or better utilization of existing resources, such as side-streams or underutilized resources to maintain a sustainable fishery. Starfish has been, until recently, an unexploited resource with limited utilization. Currently, starfish is processed into starfish meal for feed. However, the content of bioactive compounds, such as omega-3 fatty acids and phospholipids, could make it a new source of marine oil containing omega-3 fatty acids for human consumption. The aim of this study was to map the composition of bioactive compounds in starfish and starfish meal at different harvesting times to elucidate the content and variation over seasons. The results showed that starfish is a good source of marine omega-3 fatty acids and rich in phospholipids. Some variation was observed in the composition, especially for EPA bound to phospholipids, which was significantly higher in the spring. Traditional extraction using heat and mechanical separation was not applicable to the starfish, and neither was enzyme-assisted extraction. On the other hand, the supercritical CO2 extraction method using EtOH as a co-solvent seemed to be a promising green technology for extracting not only non-polar lipids, but also polar lipids, such as phospholipids. However, the conditions for extraction need further optimization.

Fish Liver Discards as a Source of Long-Chain Omega-3 Polyunsaturated Fatty Acids.

The intake of omega-3 polyunsaturated fatty acids (PUFA) of the average consumer is generally low, and products such as fish oils high in omega-3 PUFA have become popular dietary supplements. There is a need for more sources of omega-3 PUFA to cover the increasing demand. This study investigated whether livers from different lean fish species could be a potential new source of oils rich in omega-3 PUFA. The seasonal variation in lipid content, fatty acid composition, peroxide value and free fatty acid content (FFA) of livers from cod, hake, ling, coalfish and monkfish was determined, and the effect of storage conditions on the fishing vessel (ice vs frozen) was studied. Generally, the lipid content and composition of the livers from the five fish species varied similarly during the two years of the sampling period, with significantly lower values in spring (March, April) and higher values in fall (October, November). Storage conditions were found to have no significant effect on the quality and oil composition. Monkfish livers were less suitable for production of omega-3 oil due to their lower lipid and EPA content as well as higher FFA levels. Coalfish had higher fluctuations in oil composition during the sampling period, which potentially makes a standardised quality difficult to obtain. Cod, hake and ling were the most suitable species for fish liver oil production.

Biochemical Characterization and Storage Stability of Process Waters from Industrial Shrimp Production.

Shrimp boiling water (SBW) and shrimp peeling water (SPW), generated during shrimp processing, were characterized in terms of crude composition, volatile compounds, as well as nutritional and potentially toxic elements over a 13 month sampling period. The storage stability of both waters was also evaluated. Results showed that SBW contained on median 14.8 g/L protein and 2.2 g/L total fatty acids with up to 50% comprising eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Astaxanthin esters, which dominated the total astaxanthin, were 2.8 mg/L on median. SPW, on the other hand, contained on median 1.0 g/L of protein, 0.21 g/L of total fatty acids, and 1.2 mg/L astaxanthin esters. For both side-streams, essential amino acids were up to 50% of total amino acids. For SBW and SPW, the most abundant nutritional elements were Na, K, P, Ca, Cu, and Zn. The contents of all potentially toxic elements were below the detection limits, except for As. SBW was more stable at 4 °C compared to SPW as shown, e.g., by thiobarbituric acid reactive substances and relative changes in total volatile basic nitrogen. The extensive compositional mapping of SBW/SPW provides crucial knowledge necessary in the exploitation and value-adding of such side-streams into food or feed products.

Biofunctionality of Enzymatically Derived Peptides from Codfish (Gadus morhua) Frame: Bulk In Vitro Properties, Quantitative Proteomics, and Bioinformatic Prediction.

Protein hydrolysates show great promise as bioactive food and feed ingredients and for valorization of side-streams from e.g., the fish processing industry. We present a novel approach for hydrolysate characterization that utilizes proteomics data for calculation of weighted mean peptide properties (length, molecular weight, and charge) and peptide-level abundance estimation. Using a novel bioinformatic approach for subsequent prediction of biofunctional properties of identified peptides, we are able to provide an unprecedented, in-depth characterization. The study further characterizes bulk emulsifying, foaming, and in vitro antioxidative properties of enzymatic hydrolysates derived from cod frame by application of Alcalase and Neutrase, individually and sequentially, as well as the influence of heat pre-treatment. All hydrolysates displayed comparable or higher emulsifying activity and stability than sodium caseinate. Heat-treatment significantly increased stability but showed a negative effect on the activity and degree of hydrolysis. Lower degrees of hydrolysis resulted in significantly higher chelating activity, while the opposite was observed for radical scavenging activity. Combining peptide abundance with bioinformatic prediction, we identified several peptides that are likely linked to the observed differences in bulk emulsifying properties. The study highlights the prospects of applying proteomics and bioinformatics for hydrolysate characterization and in food protein science.

The effect of rosemary (Rosmarinus officinalis L.) extract on the oxidative stability of lipids in cow and soy milk enriched with fish oil.

Lipid oxidation of fish oil enriched cow milk and soy milk supplemented with rosemary extract stored at 2 °C was studied. Both peroxide value and volatile secondary lipid oxidation products were determined to monitor the progress of lipid oxidation. Rosemary extract inhibited lipid oxidation in fish oil enriched cow milk. In contrast, soy milk samples having much higher unsaturated fatty acid content showed higher lipid oxidation stability compared to cow milk. Reduction in the content of chlorogenic acid during storage suggested that this compound may contribute to the lipid oxidation stability of fish oil enriched soy milk product. Total carnosic acid and carnosol concentration declined much faster in soy milk than in cow milk. It is suggested from the results that food components could have significant impact on the fate of bioactive antioxidant compounds in a specific food product during storage.

Combination of sodium caseinate and succinylated alginate improved stability of high fat fish oil-in-water emulsions.

Sodium caseinate (CAS) and commercial sodium alginate (CA), long chain modified alginate (LCMA) or short chain modified alginate (SCMA) were used in combination for emulsifying and stabilizing high fat (50-70%) fish oil-in-water emulsions. Physical (creaming, droplet size, viscosity and protein determination) and oxidative (primary and secondary oxidation products) stabilities of the emulsions were studied during 12 days of storage. Creaming stability was higher for emulsions produced with alginates and CAS compared to emulsions prepared with only CAS. Combined use of CAS + LCMA performed better in terms of physical stability compared to emulsions produced with only CAS. However, the oxidative stability of this emulsion was inferior probably due to the presence of an unsaturated carbon chain in LCMA structure. CAS + SCMA emulsions not only showed better physical stability such as smaller droplet size, lower creaming and higher viscosity, but also had an improved oxidative stability than emulsions produced with only CAS.

Effects of Different Lipophilized Ferulate Esters in Fish Oil-Enriched Milk: Partitioning, Interaction, Protein, and Lipid Oxidation.

Antioxidant effects of ferulic acid and lipophilized ferulate esters were investigated in fish oil-enriched milk. Methyl ferulate (C1) and ethyl ferulate (C2) more efficiently prevented lipid oxidation than dodecyl ferulate (C12) did, followed by ferulic acid (C0). The combination of C1 or C2 with C12 could have a "synergistic" effect indicated by peroxide value, hexanal, and 1-penten-3-ol analysis results. These antioxidants also showed protein oxidation inhibition effects. The most effective antioxidants (C1 and C2) had the highest concentration in the precipitate phase but the lowest concentration in the aqueous phase, which was the opposite of the partitioning of C0. C12 had the highest concentration in the oil and emulsion phase. In particular, the interaction between ferulates esterified with short and medium alkyl chain lengths could lead to their "synergistic" effects in fish oil-enriched milk, which could be caused by the change in their partitioning or localization at the interface.

Physical and oxidative stability of fish oil-in-water emulsions fortified with enzymatic hydrolysates from common carp (Cyprinus carpio) roe.

Physical and oxidative stability of 5% (by weight) cod liver oil-in-water emulsions fortified with common carp (C. carpio) roe protein hydrolysate (CRPH) were examined. CRPH was obtained by enzymatic hydrolysis of discarded roe by using Alcalase 2.4L for 30, 60, 90, and 120min to yield different degrees of hydrolysis (DH). All the hydrolysates showed in vitro antioxidant activity in terms of radical scavenging and chelating properties. CRPH-containing emulsions had significantly smaller droplets than control (p<0.05). Besides, CRPH rendered rheological stability to the emulsions. It also prevented the loss of tocopherol and polyunsaturated fatty acids (PUFAs), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). CRPH retarded primary and secondary oxidation in emulsions as evidenced by peroxide values (PVs) and secondary volatile oxidation products, respectively. All the mentioned effects were compared among CRPHs with varying DH (7.6-10.2%). However, CRPH-containing emulsions had high levels of 2-methyl-1-butanol, 3-methyl-1-butanol, and 2-butanone after storage.

Antioxidative effect of lipophilized caffeic acid in fish oil enriched mayonnaise and milk.

The antioxidative effect of lipophilized caffeic acid was assessed in two different fish oil enriched food products: mayonnaise and milk. In both emulsion systems, caffeic acid esterified with fatty alcohols of different chain lengths (C1-C20) were better antioxidants than the original phenolic compound. The optimal chain length with respect to protection against oxidation was, however, different for the two food systems. Fish oil enriched mayonnaise with caffeates of medium alkyl chain length (butyl, octyl and dodecyl) added resulted in a better oxidative stability than caffeates with shorter (methyl) or longer (octadecyl) alkyl chains. Whereas in fish oil enriched milk emulsions the most effective caffeates were those with shorter alkyl chains (methyl and butyl) rather than the ones with medium and long chains (octyl, dodecyl, hexadecyl and eicosyl). These results demonstrate that there might be an optimum alkyl chain length for each phenolipid in each type of emulsion systems.

Antioxidant properties and efficacies of synthesized alkyl caffeates, ferulates, and coumarates.

Caffeic, ferulic, and coumaric acids were lipophilized with saturated fatty alcohols (C1-C20). The antioxidant properties of these hydroxycinnamic acids and their alkyl esters were evaluated in various assays. Furthermore, the antioxidant efficiency of the compounds was evaluated in a simple o/w microemulsion using the conjugated autoxidizable triene (CAT) assay. All evaluated phenolipids had radical scavenging, reducing power, and metal chelating properties. Only caffeic acid and caffeates were able to form a complex with iron via their catechol group in the phenolic ring. In the o/w emulsion, the medium chain phenolipids of the three homologues series were most efficient. The antioxidant properties and efficacies were dependent upon functional groups substituted to the ring structure and were in the following order: caffeic acid and caffeates > ferulic acid and ferulates > coumaric acid and coumarates. Moreover, the results demonstrated that the test system has an impact on the antioxidative properties measured.

Interactions between iron, phenolic compounds, emulsifiers, and pH in omega-3-enriched oil-in-water emulsions.

The behavior of antioxidants in emulsions is influenced by several factors such as pH and emulsifier type. This study aimed to evaluate the interaction between selected food emulsifiers, phenolic compounds, iron, and pH and their effect on the oxidative stability of n-3 polyunsaturated lipids in a 10% oil-in-water emulsion. The emulsifiers tested were Tween 80 and Citrem, and the phenolic compounds were naringenin, rutin, caffeic acid, and coumaric acid. Lipid oxidation was evaluated at all levels, that is, formation of radicals (ESR), hydroperoxides (PV), and secondary volatile oxidation products. When iron was present, the pH was crucial for the formation of lipid oxidation products. At pH 3 some phenolic compounds, especially caffeic acid, reduced Fe(3+) to Fe(2+), and Fe(2+) increased lipid oxidation at this pH compared to pH 6. Among the evaluated phenols, caffeic acid had the most significant effects, as caffeic acid was found to be prooxidative irrespective of pH, emulsifier type, and presence of iron, although the degrees of lipid oxidation were different at the different experimental conditions. The other evaluated phenols were prooxidative at pH 3 in Citrem-stabilized emulsions and had no significant effect at pH 6 in Citrem- or Tween-stabilized emulsions on the basis of the formation of volatiles. The results indicated that phenol-iron complexes/nanoparticles were formed at pH 6.