Purification and Molecular Characterization of Fucoidan Isolated from Ascophyllum nodosum Brown Seaweed Grown in Ireland.

The present study investigates the molecular characteristics of fucoidan obtained from the brown Irish seaweed Ascophyllum nodosum, employing hydrothermal-assisted extraction (HAE) followed by a three-step purification protocol. The dried seaweed biomass contained 100.9 mg/g of fucoidan, whereas optimised HAE conditions (solvent, 0.1N HCl; time, 62 min; temperature, 120 °C; and solid to liquid ratio, 1:30 (w/v)) yielded 417.6 mg/g of fucoidan in the crude extract. A three-step purification of the crude extract, involving solvents (ethanol, water, and calcium chloride), molecular weight cut-off filter (MWCO; 10 kDa), and solid-phase extraction (SPE), resulted in 517.1 mg/g, 562.3 mg/g, and 633.2 mg/g of fucoidan (p < 0.05), respectively. In vitro antioxidant activity, as determined by 1,1-diphenyl-2-picryl-hydrazyl radical scavenging and ferric reducing antioxidant power assays, revealed that the crude extract exhibited the highest antioxidant activity compared to the purified fractions, commercial fucoidan, and ascorbic acid standard (p < 0.05). The molecular attributes of biologically active fucoidan-rich MWCO fraction was characterised by quadruple time of flight mass spectrometry and Fourier-transform infrared (FTIR) spectroscopy. The electrospray ionisation mass spectra of purified fucoidan revealed quadruply ([M+4H]4+) and triply ([M+3H]3+) charged fucoidan moieties at m/z 1376 and m/z 1824, respectively, and confirmed the molecular mass 5444 Da (~5.4 kDa) from multiply charged species. The FTIR analysis of both purified fucoidan and commercial fucoidan standard exhibited O-H, C-H, and S=O stretching which are represented by bands at 3400 cm-1, 2920 cm-1, and 1220-1230 cm-1, respectively. In conclusion, the fucoidan recovered from HAE followed by a three-step purification process was highly purified; however, purification reduced the antioxidant activity compared to the crude extract.

Nutraceuticals from Algae: Current View and Prospects from a Research Perspective.

In recent years, algae, both microalgae and macroalgae, have attracted the attention of the scientific community as a source of multiple active molecules or bioactives, including polysaccharides, fatty acids, proteins and peptides, polyphenols, diterpenes, steroids, and alkaloids [...].

Bioactive Peptides from Algae: Traditional and Novel Generation Strategies, Structure-Function Relationships, and Bioinformatics as Predictive Tools for Bioactivity.

Over the last decade, algae have been explored as alternative and sustainable protein sources for a balanced diet and more recently, as a potential source of algal-derived bioactive peptides with potential health benefits. This review will focus on the emerging processes for the generation and isolation of bioactive peptides or cryptides from algae, including: (1) pre-treatments of algae for the extraction of protein by physical and biochemical methods; and (2) methods for the generation of bioactive including enzymatic hydrolysis and other emerging methods. To date, the main biological properties of the peptides identified from algae, including anti-hypertensive, antioxidant and anti-proliferative/cytotoxic effects (for this review, anti-proliferative/cytotoxic will be referred to by the term anti-cancer), assayed in vitro and/or in vivo, will also be summarized emphasizing the structure-function relationship and mechanism of action of these peptides. Moreover, the use of in silico methods, such as quantitative structural activity relationships (QSAR) and molecular docking for the identification of specific peptides of bioactive interest from hydrolysates will be described in detail together with the main challenges and opportunities to exploit algae as a source of bioactive peptides.

Influence of Enzymatic Hydrolysis and Molecular Weight Fractionation on the Antioxidant and Lipase / α-Amylase Inhibitory Activities In Vitro of Watermelon Seed Protein Hydrolysates.

This study aims to evaluate the potential in vitro antioxidant and anti-obesity activities of watermelon seed protein hydrolysates (WSPH) obtained using different combinations of enzymes alcalase−proteinase K (ALC-PK) and alcalase−actinidin (ALC-ACT). There was a direct relationship between the degree of hydrolysis (DH) and the biological activities of the WSPH, with the highest DPPH (approximately 85%) and lipase inhibitory activities (≈59%) appreciated at DH of 36−37% and 33−35% when using ALC-PK and ALC-ACT, respectively. Following molecular weight fractionation, the ALC-PK WSPH < 3 kDa (F1) assayed at 1 mg.mL−1 had the highest DPPH-radical scavenging (89.22%), ferrous chelating (FC) (79.83%), reducing power (RP) (A 0.51), lipase inhibitory (71.36%), and α-amylase inhibitory (62.08%) activities. The amino acid analysis of ALC-PK WSPH and its fractions revealed a relationship between the biological activity of the extracts and their composition. High contents of hydrophobic amino acids, arginine, and aromatic amino acids were related to high antioxidant, lipase inhibitory, and α-amylase inhibitory activities in the extracts, respectively. Overall, this study revealed that underutilized protein sources such as WSPH, using the appropriate combination of enzymes, could result in the generation of new ingredients and compounds with powerful antioxidant and anti-obesity activities with promising applications as nutraceuticals or functional foods.

Seasonal Variation of the Proximate Composition, Mineral Content, Fatty Acid Profiles and Other Phytochemical Constituents of Selected Brown Macroalgae.

The main objective was to determine the chemical, phytochemical, fatty acid and mineral profiles of three commercially relevant brown macroalgae (Laminaria digitata, Laminaria hyperborea and Ascophyllum nodosum) collected each season for two years off the west coast of Ireland. All the chemical, phytochemical, fatty acid and minerals analysed varied significantly depending on the macroalgal species, season and year of collection. Overall, the protein contents of macroalgae were negatively correlated with carbohydrate content. Protein (2-11%) was at its highest during winter and/or spring, decreasing to a minimum during summer and/or autumn. The three macroalgal species analysed in this study had clearly differentiated fatty acid profiles. The concentration of fatty acids was higher in A. nodosum compared with both Laminaria species. The mineral profile of the three macroalgal species was rich in essential metals, particularly Ca, Mg and P, while the levels of I were approximately 9- to 10-fold higher in both Laminaria spp. compared with A. nodosum. The levels of toxic metals (Cd, Hg and Pb) in all the macroalgal species studied were low in the current study; while the levels of total As were high (49-64 mg/kg DW macroalgae) compared with previous reports.

Antimicrobials from Seaweeds for Food Applications.

The exponential growth of emerging multidrug-resistant microorganisms, including foodborne pathogens affecting the shelf-life and quality of foods, has recently increased the needs of the food industry to search for novel, natural and eco-friendly antimicrobial agents. Macroalgae are a bio-diverse group distributed worldwide, known to produce multiple compounds of diverse chemical nature, different to those produced by terrestrial plants. These novel compounds have shown promising health benefits when incorporated into foods, including antimicrobial properties. This review aims to provide an overview of the general methods and novel compounds with antimicrobial properties recently isolated and characterized from macroalgae, emphasizing the molecular pathways of their antimicrobial mechanisms of action. The current scientific evidence on the use of macroalgae or macroalgal extracts to increase the shelf-life of foods and prevent the development of foodborne pathogens in real food products and their influence on the sensory attributes of multiple foods (i.e., meat, dairy, beverages, fish and bakery products) will also be discussed, together with the main challenges and future trends of the use of marine natural products as antimicrobials.

Evaluation of the Antibacterial and Prebiotic Potential of Ascophyllum nodosum and Its Extracts Using Selected Bacterial Members of the Pig Gastrointestinal Microbiota.

Ascophyllum nodosum and its extracts are promising antibacterial and prebiotic dietary supplements for pigs. The objectives of this study were to evaluate the effects of the increasing concentrations of: (1) two whole biomass samples of A. nodosum with different harvest seasons, February (ANWB-F) and November (ANWB-N), in a weaned pig faecal batch fermentation assay, and (2) A. nodosum extracts produced using four different extraction conditions of a hydrothermal-assisted extraction methodology (ANE1-4) and conventional extraction methods with water (ANWE) and ethanol (ANEE) as solvent in individual pure culture growth assays using a panel of beneficial and pathogenic bacterial strains. In the batch fermentation assay, ANWB-F reduced Bifidobacterium spp. counts (p < 0.05) while ANWB-N increased total bacterial counts and reduced Bifidobacterium spp. and Enterobacteriaceae counts (p < 0.05). Of the ANE1-4, produced from ANWB-F, ANWE and ANEE that were evaluated in the pure culture growth assays, the most interesting extracts were the ANE1 that reduced Salmonella Typhimurium, enterotoxigenic Escherichia coli and B. thermophilum counts and the ANE4 that stimulated B. thermophilum growth (p < 0.05). In conclusion, the extraction method and conditions influenced the bioactivities of the A. nodosum extracts with ANE1 and ANE4 exhibiting distinct antibacterial and prebiotic properties in vitro, respectively, that merit further exploration.

Evaluation of Ultrasound, Microwave, Ultrasound-Microwave, Hydrothermal and High Pressure Assisted Extraction Technologies for the Recovery of Phytochemicals and Antioxidants from Brown Macroalgae.

This study aims to explore novel extraction technologies (ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), ultrasound-microwave-assisted extraction (UMAE), hydrothermal-assisted extraction (HAE) and high-pressure-assisted extraction (HPAE)) and extraction time post-treatment (0 and 24 h) for the recovery of phytochemicals and associated antioxidant properties from Fucus vesiculosus and Pelvetia canaliculata. When using fixed extraction conditions (solvent: 50% ethanol; extraction time: 10 min; algae/solvent ratio: 1/10) for all the novel technologies, UAE generated extracts with the highest phytochemical contents from both macroalgae. The highest yields of compounds extracted from F. vesiculosus using UAE were: total phenolic content (445.0 ± 4.6 mg gallic acid equivalents/g), total phlorotannin content (362.9 ± 3.7 mg phloroglucinol equivalents/g), total flavonoid content (286.3 ± 7.8 mg quercetin equivalents/g) and total tannin content (189.1 ± 4.4 mg catechin equivalents/g). In the case of the antioxidant activities, the highest DPPH activities were achieved by UAE and UMAE from both macroalgae, while no clear pattern was recorded in the case of FRAP activities. The highest DPPH scavenging activities (112.5 ± 0.7 mg trolox equivalents/g) and FRAP activities (284.8 ± 2.2 mg trolox equivalents/g) were achieved from F. vesiculosus. Following the extraction treatment, an additional storage post-extraction (24 h) did not improve the yields of phytochemicals or antioxidant properties of the extracts.

Exploring Ultrasound, Microwave and Ultrasound-Microwave Assisted Extraction Technologies to Increase the Extraction of Bioactive Compounds and Antioxidants from Brown Macroalgae.

This study aims to determine the influence of (1) ultrasound-assisted extraction (UAE), (2) microwave-assisted extraction (MAE) and (3) a combination of ultrasound-microwave-assisted extraction (UMAE) on the yields of fucose-sulphated polysaccharides (FSPs), total soluble carbohydrates and antioxidants extracted from A. nodosum. Scanning electron microscopy (SEM) was used to evaluate the influence of the extraction technologies on the surface of macroalgae while principal component analysis was used to assess the influence of the extraction forces on the yields of compounds. UMAE generated higher yields of compounds compared to UAE and MAE methods separately. The maximum yields of compounds achieved using UMAE were: FSPs (3533.75 ± 55.81 mg fucose/100 g dried macroalgae (dm)), total soluble carbohydrates (10408.72 ± 229.11 mg glucose equivalents/100 g dm) and phenolic compounds (2605.89 ± 192.97 mg gallic acid equivalents/100 g dm). The antioxidant properties of the extracts showed no clear trend or extreme improvements by using UAE, MAE or UMAE. The macroalgal cells were strongly altered by the application of MAE and UMAE, as revealed by the SEM images. Further research will be needed to understand the combined effect of sono-generated and microwave-induced modifications on macroalgae that will allow us to tailor the forces of extraction to target specific molecules.

Optimisation of Ultrasound Frequency, Extraction Time and Solvent for the Recovery of Polyphenols, Phlorotannins and Associated Antioxidant Activity from Brown Seaweeds.

This study investigates ultrasound assisted extraction (UAE) process parameters (time, frequency and solvent) to obtain high yields of phlorotannins, flavonoids, total phenolics and associated antioxidant activities from 11 brown seaweed species. Optimised UAE conditions (35 kHz, 30 min and 50% ethanol) significantly improved the extraction yield from 1.5-fold to 2.2-fold in all seaweeds investigated compared to solvent extraction. Using ultrasound, the highest recovery of total phenolics (TPC: 572.3 ± 3.2 mg gallic acid equivalent/g), total phlorotannins (TPhC: 476.3 ± 2.2 mg phloroglucinol equivalent/g) and total flavonoids (TFC: 281.0 ± 1.7 mg quercetin equivalent/g) was obtained from Fucus vesiculosus seaweed. While the lowest recovery of TPC (72.6 ± 2.9 mg GAE/g), TPhC (50.3 ± 2.0 mg PGE/g) and TFC (15.2 ± 3.3 mg QE/g) was obtained from Laminaria digitata seaweed. However, extracts from Fucus serratus obtained by UAE exhibited the strongest 1,1-diphenyl-2-picryl-hydrazyl (DPPH) scavenging activity (29.1 ± 0.25 mg trolox equivalent/g) and ferric reducing antioxidant power (FRAP) value (63.9 ± 0.74 mg trolox equivalent/g). UAE under optimised conditions was an effective, low-cost and eco-friendly technique to recover biologically active polyphenols from 11 brown seaweed species.

In Vitro and In Silico Approaches to Generating and Identifying Angiotensin-Converting Enzyme I Inhibitory Peptides from Green Macroalga Ulva lactuca.

A protein extract was generated from the macroalga Ulva lactuca, which was subsequently hydrolysed using the food-grade enzyme papain and angiotensin-converting Enzyme I and renin inhibitory peptides identified using a combination of enrichment strategies employing molecular weight cutoff filtration and mass spectrometry analysis. The generated hydrolysates with the most promising in vitro activity were further purified using preparative RP-HPLC and characterised. The 1 kDa hydrolysate (1 kDa-UFH), purified and collected by preparative RP-HPLC at minutes 41‒44 (Fr41‒44), displayed statistically higher ACE-I inhibitory activities ranging from 96.91% to 98.06%. A total of 48 novel peptides were identified from these four fractions by LC-MS/MS. A simulated gastrointestinal digestion of the identified peptide sequences was carried out using in silico enzyme cleavage simulation tools, resulting in 86 peptide sequences that were further assessed for their potential activity, toxicity and allergenicity using multiple predictive approaches. All the peptides obtained in this study were predicted to be non-toxic. However, 28 out of the 86 novel peptides released after the in silico gastrointestinal digestion were identified as potential allergens. The potential allergenicity of these peptides should be further explored to comply with the current labelling regulations in formulated food products containing U. lactuca protein hydrolysates.

Enhancing the Extraction of Polysaccharides and Antioxidants from Macroalgae Using Sequential Hydrothermal-Assisted Extraction Followed by Ultrasound and Thermal Technologies.

Fucose sulphated polysaccharides (FSPs) and glucans have recently attracted the attention of the scientific community due to their wide range of biological activities. Both polysaccharides should ideally be selectively extracted using innovative technologies with high extraction efficiency. This study aims to: (1) Optimise the extraction variables used in hydrothermal-assisted extraction (HAE) to obtain high yields of FSPs, total glucans, and antioxidants from Laminaria hyperborea; (2) to apply these optimised protocols to other brown macroalgae; and (3) to explore the application of ultrasound and thermal technologies to increase the recovery of polysaccharides from the residual biomass. Box-Behnken design (three-factor, four-levels) was employed to optimise the HAE variables, and principal component analysis was used to evaluate the recovery of polysaccharides from the residual biomass. The optimal HAE conditions were 120 °C, 80.9 min, and 12.02 mL/g macroalgae from L. hyperborea. The best sequential application of ultrasound and thermal treatment achieved an additional 2971.7 ± 61.9 mg fucose/100 g dried macroalgal residue (dmr) from Ascophyllum nodosum and 908.0 ± 51.4 mg total glucans/100 g dmr from L. hyperborea macroalgal residues.

Extraction and Yield Optimisation of Fucose, Glucans and Associated Antioxidant Activities from Laminaria digitata by Applying Response Surface Methodology to High Intensity Ultrasound-Assisted Extraction.

The objectives of this study were to employ response surface methodology (RSM) to investigate and optimize the effect of ultrasound-assisted extraction (UAE) variables, temperature, time and amplitude on the yields of polysaccharides (fucose and total glucans) and antioxidant activities (ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picryl-hydrazyl radical scavenging activity (DPPH)) from Laminariadigitata, and to explore the suitability of applying the optimum UAE conditions for L.digitata to other brown macroalgae (L.hyperborea and Ascophyllumnodosum). The RSM with three-factor, four-level Box-Behnken Design (BBD) was used to study and optimize the extraction variables. A second order polynomial model fitted well to the experimental data with R² values of 0.79, 0.66, 0.64, 0.73 for fucose, total glucans, FRAP and DPPH, respectively. The UAE parameters studied had a significant influence on the levels of fucose, FRAP and DPPH. The optimised UAE conditions (temperature = 76 °C, time = 10 min and amplitude = 100%) achieved yields of fucose (1060.7 ± 70.6 mg/100 g dried seaweed (ds)), total glucans (968.6 ± 13.3 mg/100 g ds), FRAP (8.7 ± 0.5 µM trolox/mg freeze-dried extract (fde)) and DPPH (11.0 ± 0.2%) in L.digitata. Polysaccharide rich extracts were also attained from L.hyperborea and A. nodosum with variable results when utilizing the optimum UAE conditions for L.digitata.

Algae-derived compounds: Bioactivity, allergenicity and technologies enhancing their values.

As a rapidly growing source of human nutrients, algae biosynthesize diverse metabolites which have promising bioactivities. However, the potential allergenicity of algal components hinder their widespread adoption. This review provides a comprehensive review of various macro and micronutrients derived from algal biomass, with particular focus on bioactive compounds, including peptides, polyphenols, carotenoids, omega-3 fatty acids and phycocyanins. The approaches used to produce algal bioactive compounds and their health benefits (antioxidant, antidiabetic, cardioprotective, anti-inflammatory and immunomodulatory) are summarised. This review particularly focuses on the state-of-the-art of precision fermentation, encapsulation, cold plasma, high-pressure processing, pulsed electric field, and subcritical water to reduce the allergenicity of algal compounds while increasing their bioactivity and bioavailability. By providing insights into current challenges of algae-derived compounds and opportunities for advancement, this review contributes to the ongoing discourse on maximizing their application potential in the food nutraceuticals, and pharmaceuticals industries.

Spray-drying and rehydration on ß-carotene encapsulated Pickering emulsion with chitosan and seaweed polyphenol.

The spray-drying process to generate microcapsules from Pickering emulsions needs high temperatures, leading to instability of emulsions and degradation of encapsulated thermosensitive compounds (ß-carotene). However, these effects may be attenuated by the introduction of seaweed polyphenols into the emulsion interfacial layers, although the effects underlying this protective mechanism have not been explored. This study evaluates the effects of spray-drying/rehydration on the morphology, encapsulation efficiency, redispersibility, and stability of ß-carotene loaded Pickering emulsions stabilized by chitosan (PESC) and Pickering emulsions stabilized by chitosan/seaweed polyphenols (PESCSP). The encapsulation efficiency of ß-carotene in PESCSP microcapsules (61.13 %) was higher than PESC (53.91 %). Rehydrated PESCSP exhibited more regular droplet size distribution, higher stability, stronger 3D network morphology, and lower redispersibility index (1.5) compared to rehydrated PESC. Analyses of interfacial layers of emulsions revealed that chitosan covalently bound fatty acids at their hydrophobic side. Polyphenols were linked to chitosan at the hydrophilic side of emulsions through hydrogen bonds, providing 3D network between droplets and antioxidant activities to inhibit the degradation of ß-carotene. This study emphasized the role of polyphenols in the interfacial layers of Pickering emulsions for the development of efficient delivery systems and protection of ß-carotene and other thermosensitive bioactive compounds during spray-drying and rehydration.

Acoustic cavitation for agri-food applications: Mechanism of action, design of new systems, challenges and strategies for scale-up.

Acoustic cavitation, an intriguing phenomenon resulting from the interaction of sound waves with a liquid medium, has emerged as a promising avenue in agri-food processing, offering opportunities to enhance established processes improving primary production of ingredients and further food processing. This comprehensive review provides an in-depth analysis of the mechanisms, design considerations, challenges and scale-up strategies associated with acoustic cavitation for agri-food applications. The paper starts by elucidating the fundamental principles of acoustic cavitation and its measurement, delving then into the diverse effects of different parameters associated with, the acoustic wave, mechanical design and operation of the ultrasonic system, along with those related to the food matrix. The technological advancements achieved in the design and set-up of ultrasonic reactors addressing limitations during scale up are also discussed. The design, engineering and mathematical modelling of ultrasonic equipment tailored for agri-food applications are explored, along with strategies to maximize cavitation intensity and efficiency in the application of brining, freezing, drying, emulsification, filtration and extraction. Advanced US equipment, such as multi-transducers (tubular resonator, FLOW:WAVE®) and larger processing surface areas through innovative designing (Barbell horn, CascatrodesTM), are one of the most promising strategies to ensure consistency of US operations at industrial scale. This review paper aims to provide valuable insights into harnessing acoustic cavitation's potential for up-scaling applications in food processing via critical examination of current research and advancements, while identifying future directions and opportunities for further research and innovation.

A review on proteomic and genomic biomarkers for gelatin source authentication: Challenges and future outlook.

Biomarkers are compounds that could be detected and used as indicators of normal and/or abnormal functioning of different biological systems, including animal tissues and food matrices. Gelatin products of animal origin, mainly bovine and porcine, are currently under scrutiny mainly due to the specific needs of some sectors of the population related to religious beliefs and their dietary prohibitions, as well as some potential health threats associated with these products. Thus, manufacturers are currently in need of a reliable, convenient, and easy procedure to discern and authenticate the origin of animal-based gelatins (bovine, porcine, chicken, or fish). This work aims to review current advances in the creation of reliable gelatin biomarkers for food authentication purposes based on proteomic and DNA biomarkers that could be applied in the food sector. Overall, the presence of specific proteins and peptides in gelatin can be chemically analysed (i.e., by chromatography, mass spectroscopy, electrophoresis, lateral flow devices, and enzyme-linked immunosorbent assay), and different polymerase chain reaction (PCR) methods have been applied for the detection of nucleic acid substances in gelatin. Altogether, despite the fact that numerous methods are currently being developed for the purpose of detecting gelatin biomarkers, their widespread application is highly dependent on the cost of the equipment and reagents as well as the ease of use of the various methods. Combining different methods and approaches targeting multiple biomarkers may be key for manufacturers to achieve reliable authentication of gelatin's origin.

Pickering emulsions with chitosan and macroalgal polyphenols stabilized by layer-by-layer electrostatic deposition.

Due to the hydrophilic nature of chitosan, its use as stabilizers in Pickering emulsions requires of hydrophobic modifications that can be achieved by layer-by-layer electrostatic deposition. This study aims to explore and optimize the effects of multiple parameters (chitosan-polyphenol ratio, chitosan concentrations, oil-phase ratios, pH and NaCl concentrations) on the stability of Pickering emulsions stabilized by chitosan and macroalgal polyphenols. The binding mode of chitosan and polyphenols in co-stabilized Pickering emulsions was characterized by Fourier transform infrared spectroscopy (FTIR), confocal scanning laser microscopy (CLSM), scanning electron microscopy (SEM), atomic force microscopy (AFM), and contact angle. The emulsions had the highest physical stability (95.67 %) and specific surface area (2.21 m2/g) at: chitosan-polyphenol ratio 9.5:1 (w:w), chitosan concentration 0.5 % (w/v), oil-phase ratio 5 % (v/v), pH 6, and NaCl concentration 0 mM. FTIR, CLSM, SEM, and AFM evidenced that chitosan and polyphenols were bound by hydrogen bonds and electrostatic interactions at oil-water interfacial layers of emulsions that had a contact angle of 87.8°, indicating that polyphenols combined with chitosan had better ability to stabilize Pickering emulsions compared to chitosan alone. This study broadens the potential applications of naturally extracted macroalgal polyphenols for the development of Pickering emulsions in nutraceutical and functional food industries.

Effects of environmental stimuli on the physicochemical and rheological properties of chitosan-macroalgal polyphenol stabilized Pickering emulsion.

In this study, Pickering emulsions stabilized by chitosan (CS), chitosan-Laminaria japonica polyphenols (CP) and chitosan-Ascophyllum nodosum polyphenols (CB) were fabricated. This study also evaluated the stability of CS, CP, and CB under different environmental factors including pH (2-9), NaCl concentrations (0-500 mM), heat treatments (50-100 °C) and storage period (0-8 weeks). The characterization on interfacial layer of emulsion droplets demonstrated that macroalgal polyphenols could combined with the amorphous regions of chitosan particles through hydrogen bond and electrostatic interactions, providing stronger dual wettability with enhanced ability of interfacial layer in stabilizing Pickering emulsions. All three emulsions showed best droplet distribution, highest emulsion stability and specific surface area at pH 6 and 0 mM NaCl concentration as fresh emulsion. Moreover, CS, CP, and CB exhibited the rheological behaviour of pseudoplastic fluids at different pH and NaCl concentration. It should be noted that CP and CB exhibited higher emulsion stability than CS under a variety of environmental stresses. Overall, this research proved that chitosan-macroalgal polyphenol co-stabilized Pickering emulsion had enhanced stability against various environmental stimuli, which could be utilized as potential delivery and protection system for hydrophobic bioactive compounds.

Chitosan-based Pickering emulsion: A comprehensive review on their stabilizers, bioavailability, applications and regulations.

BACKGROUND: Chitosan-based particles are one of the most promising Pickering emulsions stabilizers due to its cationic properties, cost-effective, biocompatibility, biodegradability. However, there are currently no comprehensive reviews analyzing the role of chitosan to develop Pickering emulsions, and the bioavailability and multiple uses of these emulsions. SCOPE AND APPROACH: This review firstly summarizes the types, preparation and functional properties of chitosan-based Pickering emulsion stabilizers, followed by in vivo and in vitro bioavailability, main regulations, and future application and trends. KEY FINDINGS AND CONCLUSIONS: Stabilizers used in chitosan-based Pickering emulsions include 6 categories: chitosan self-aggregating particles and 5 types of composites (chitosan-protein, chitosan-polysaccharide, chitosan-fatty acid, chitosan-polyphenol, and chitosan-inorganic). Chitosan-based Pickering emulsions improved the bioavailability of different compounds compared to traditional emulsions. Current applications include hydrogels, microcapsules, food ingredients, bio-based films, cosmeceuticals, porous scaffolds, environmental protection agents, and interfacial catalysis systems. However, due to current limitations, more research and development are needed to be extensively explored to meet consumer demand, industrial manufacturing, and regulatory requirements. Thus, optimization of stabilizers, bioavailability studies, 3D4D printing, fat substitutes, and double emulsions are the main potential development trends or research gaps in the field which would contribute to increase adoption of these promising emulsions at industrial level.