Sustainable Extraction Protocols for the Recovery of Bioactive Compounds from By-Products of Pomegranate Fruit Processing.

This study investigates sustainable extraction protocols for the recovery of bioactive compounds from by-products of various pomegranate (Punica granatum L.) cultivars, including Acco, Hicaz, Jolly Red, Parfianka, Valenciana, and Wonderful, generated during the industrial processing of the fruits. Advanced extraction technologies, including ultrasounds, microwaves, and hydrodynamic cavitation, have been compared to conventional extraction procedures and utilized to enhance extraction efficiency while also minimizing environmental impact. Water-based extraction methods have been utilized to promote the development of sustainable and eco-friendly processes. The comparison between conventional extractions and ultrasound-assisted extractions (UAEs) and microwave-assisted extractions (MAEs) demonstrated notable improvements in extraction yields, particularly for ellagitannins (punicalins, punicalagins, and ellagic acid) and total polyphenols, with increases ranging from about 45 to 200%. However, the increases directly comparing UAEs to MAEs ranged from about 4 to 6%. This indicates that while both UAEs and MAEs offer notable improvements over conventional extractions, the differences in extraction efficiency between the two advanced methods were relatively modest. These advancements were observed across various pomegranate cultivars, highlighting the versatility and effectiveness of these methods. Notably, hydrodynamic cavitation-based extractions (HC) emerged as particularly promising, consistently yielding the highest levels of bioactive compounds (ellagitannins and total polyphenols), especially when operated at higher frequencies. Compared to conventional extractions, HC exhibited substantial increases in extraction yields for Wonderful pomegranate by-products, surpassing the efficiency of both UAEs and MAEs (approximately 45 and 57% for UAE and MAE, respectively, versus about 80% for HC). Among these advanced techniques, HC has emerged as particularly promising, yielding the most favorable results and leading to significant improvements in the yield of bioactive compounds. When directly compared to UAEs and MAEs, HC increased extraction yields by over 20%. Furthermore, HC allowed for shorter extraction times. The Wonderful cultivar consistently exhibited the highest levels of ellagitannins and the highest total polyphenol content among all types of extraction procedures used, whether conventional or advanced. This highlights the great potential of the Wonderful cultivar in terms of bioactive compound extraction and underscores its significance in research and applications related to pomegranate processing and utilization. This study suggests that the implementation of these advanced technologies into extraction processes represents a significant advancement in the field, offering a promising avenue for the development of efficient and environmentally friendly extraction methods for obtaining valuable bioactive compounds from pomegranate processing by-products.

Supercritical Extraction Techniques for Obtaining Biologically Active Substances from a Variety of Plant Byproducts.

Supercritical fluid extraction (SFE) techniques have garnered significant attention as green and sustainable methods for obtaining biologically active substances from a diverse array of plant byproducts. This paper comprehensively reviews the use of supercritical fluid extraction (SFE) in obtaining bioactive compounds from various plant residues, including pomace, seeds, skins, and other agricultural byproducts. The main purpose of supercritical fluid extraction (SFE) is the selective isolation and recovery of compounds, such as polyphenols, essential oils, vitamins, and antioxidants, that have significant health-promoting properties. Using supercritical carbon dioxide as the solvent, supercritical fluid extraction (SFE) not only eliminates the need for hazardous organic solvents, e.g., ethanol, and methanol, but also protects heat-sensitive bioactive compounds. Moreover, this green extraction technique contributes to waste valorisation by converting plant byproducts into value-added extracts with potential applications in the food, pharmaceutical, and cosmetic industries. This review highlights the advantages of SFE, including its efficiency, eco-friendliness, and production of residue-free extracts, while discussing potential challenges and future prospects for the utilisation of SFE in obtaining biologically active substances from plant byproducts.

Green Extraction of Polyphenols from Elaeagnus angustifolia L. Using Natural Deep Eutectic Solvents and Evaluation of Bioactivity.

In the study, natural deep eutectic solvents (NADESs) were used as alternatives to traditional chemical solvents for the extraction of polyphenols from Elaeagnus angustifolia L. Nine NADESs were tested for the first time and compared with ethanol and water (traditional solvents) regarding the extraction of phenolic compounds from E. angustifolia L. These solvents were particularly effective at extracting polyphenols, whose low water solubility usually requires high amounts of organic solvents. The solvent based on choline chloride and malonic acid provided optimal results and was selected for further optimization. The effects of material-to-liquid ratio, ultrasound time, and ultrasound temperature on the extraction efficiency were studied through single-factor experiments. These parameters were optimized by Box-Behnken design using response surface methodology. The optimal conditions identified were 49.86 g/mL of material-to-liquid ratio, 31.10 min of ultrasound time, and 62.35 °C of ultrasound temperature, resulting in a high yield of 140.30 ± 0.19 mg/g. The results indicated that the NADES extraction technique provided a higher yield than the conventional extraction process. The antioxidant activity of the extract of polyphenols from E. angustifolia L. was determined, and UPLC-IMS-QTOF-MS was used to analyze the phenolic compounds in it. The results revealed that the scavenging ability of 1,1-diphenyl-2-picryl-hydrazil and 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulphonate) extracted by NADES was higher than that of polyphenols extracted by water and ethanol. Furthermore, a total of 24 phenolic compounds were identified in the extract. To the best of our knowledge, this is the first study in which a green and efficient NADES extraction method has been used to extract bioactive polyphenols from E. angustifolia L., which could provide potential value in pharmaceuticals, cosmetics, and food additives.

Miniaturized matrix solid-phase dispersion assisted by deep eutectic solvent for acrylamide determination in bread samples.

Acrylamide determination is important to state its quantity in baked food preventing any potential carcinogenic effects. Matrix solid-phase dispersion (MSPD) extraction is an extraction procedure based on a homogenization phase between a solid sample and a solid dispersing material to break sample increasing analyte extraction yield, often used for acrylamide determination. The addition of a green deep eutectic solvent (DES) during the MSPD homogenization phase improves the analyte extraction, giving the possibility to reduce the amount of organic solvent used. In this work, a miniaturized MSPD extraction assisted by a DES was developed to determine acrylamide in bread, using high-performance liquid chromatography coupled with mass spectrometry detection. The optimized procedure provides 1:1 (w/w) matrix-to-dispersing material ratio, 2 mL of methanol as extraction solvent, and 50 µL of choline chloride-glycerol DES added during the homogenization phase. Method validation ensured good results with minimum recoveries of 90%, high precision with a maximum intra-day error of 4%, and inter-day error of 6%. Limit of detection and limit of quantification resulted to be 16 µg/kg and 35 µg/kg, respectively. This miniaturized extraction procedure represents a good alternative to those reported in the literature, guaranteeing great performance and respecting green chemistry principles.

Shrimp Waste Upcycling: Unveiling the Potential of Polysaccharides, Proteins, Carotenoids, and Fatty Acids with Emphasis on Extraction Techniques and Bioactive Properties.

Shrimp processing generates substantial waste, which is rich in valuable components such as polysaccharides, proteins, carotenoids, and fatty acids. This review provides a comprehensive overview of the valorization of shrimp waste, mainly shrimp shells, focusing on extraction methods, bioactivities, and potential applications of these bioactive compounds. Various extraction techniques, including chemical extraction, microbial fermentation, enzyme-assisted extraction, microwave-assisted extraction, ultrasound-assisted extraction, and pressurized techniques are discussed, highlighting their efficacy in isolating polysaccharides, proteins, carotenoids, and fatty acids from shrimp waste. Additionally, the bioactivities associated with these compounds, such as antioxidant, antimicrobial, anti-inflammatory, and antitumor properties, among others, are elucidated, underscoring their potential in pharmaceutical, nutraceutical, and cosmeceutical applications. Furthermore, the review explores current and potential utilization avenues for these bioactive compounds, emphasizing the importance of sustainable resource management and circular economy principles in maximizing the value of shrimp waste. Overall, this review paper aims to provide insights into the multifaceted aspects of shrimp waste valorization, offering valuable information for researchers, industries, and policymakers interested in sustainable resource utilization and waste-management strategies.

Exploring the Nutritional and Bioactive Potential of Olive Leaf Residues: A Focus on Minerals and Polyphenols in the Context of Spain's Olive Oil Production.

Lyophilized plant-origin extracts are rich in highly potent antioxidant polyphenols. In order to incorporate them into food products, it is necessary to protect these phytochemicals from atmospheric factors such as heat, light, moisture, or pH, and to enhance their bioavailability due to their low solubility. To address these challenges, recent studies have focused on the development of encapsulation techniques for antioxidant compounds within polymeric structures. In this study, lyophilized olive leaf extracts were microencapsulated with the aim of overcoming the aforementioned challenges. The method used for the preparation of the studied microparticles involves external ionic gelation carried out within a water-oil (W/O) emulsion at room temperature. HPLC analysis demonstrates a high content of polyphenols, with 90% of the bioactive compounds encapsulated. Meanwhile, quantification by inductively coupled plasma optical emission spectroscopy (ICP-OES) reveals that the dried leaves, lyophilized extract, and microencapsulated form contain satisfactory levels of macro- and micro-minerals (calcium, potassium, sodium). The microencapsulation technique could be a novel strategy to harness the polyphenols and minerals of olive leaves, thus enriching food products and leveraging the antioxidant properties of the polyphenolic compounds found in the lyophilized extract.

Statistical Tools to Optimize the Recovery of Bioactive Compounds from Marine Byproducts.

Techniques for extracting important bioactive molecules from seafood byproducts, viz., bones, heads, skin, frames, fins, shells, guts, and viscera, are receiving emphasis due to the need for better valorization. Employing green extraction technologies for efficient and quality production of these bioactive molecules is also strictly required. Hence, understanding the extraction process parameters to effectively design an applicable optimization strategy could enable these improvements. In this review, statistical optimization strategies applied for the extraction process parameters of obtaining bioactive molecules from seafood byproducts are focused upon. The type of experimental designs and techniques applied to criticize and validate the effects of independent variables on the extraction output are addressed. Dominant parameters studied were the enzyme/substrate ratio, pH, time, temperature, and power of extraction instruments. The yield of bioactive compounds, including long-chain polyunsaturated fatty acids, amino acids, peptides, enzymes, gelatine, collagen, chitin, vitamins, polyphenolic constituents, carotenoids, etc., were the most studied responses. Efficiency and/or economic and quality considerations and their selected optimization strategies that favor the production of potential bioactive molecules were also reviewed.

An Eco-Friendly Extraction and Purification Approach for Obtaining Active Ingredients for Cosmetics from Two Marine Brown Seaweeds.

Brown seaweeds are attracting attention due to their richness in bioactive compounds, in particular, their phlorotannins. We present here a case study of two Fucales, Ascophyllum nodosum and Halidrys siliquosa, sustainably collected, to produce active polyphenols for the cosmetics sector. Phenolic contents of crude extracts, obtained by Accelerated Solvent Extraction (ASE), were more elevated in H. siliquosa at 100.05 mg/g dry weight (DW) than in A. nodosum (29.51 mg/g DW), considering 3 cycles with cell inversion. The temperature of extraction for a high phenolic content and high associated antioxidant activities close to positive controls was 150 °C for both algae and the use of only one cycle was enough. A semi-purification process using Solid-phase Extraction (SPE) was carried out on both ASE crude extracts (one per species). The majority of phlorotannins were found in the ethanolic SPE fraction for A. nodosum and the hydroethanolic one for H. siliquosa. The SPE process allowed us to obtain more concentrated fractions of active phenolic compounds (×1.8 and 2 in A. nodosum and H. siliquosa, respectively). Results are discussed in regard to the exploitation of seaweeds in Brittany and to the research of sustainable processes to produce active natural ingredients for cosmetics.

Taguchi approach for assessing supercritical CO2 (sCO2) fluid extraction of polyhydroxyalkanoate (PHA) from Chlorella Vulgaris sp. microalgae.

This research explicitly investigates the utilization of Chlorella Vulgaris sp. microalgae as a renewable source for lipid production, focusing on its application in bioplastic manufacturing. This study employed the supercritical fluid extraction technique employing supercritical CO2 (sCO2) as a green technology to selectively extract and produce PHA's precursor utilizing CO2 solvent as a cleaner solvent compared to conventional extraction method. The study assessed the effects of three extraction parameters, namely temperature (40-60 °C), pressure (15-35 MPa), and solvent flow rate (4-8 ml/min). The pressure, flowrate, and temperature were found to be the most significant parameters affecting the sCO2 extraction. Through Taguchi optimization, the optimal parameters were determined as 60 °C, 35 MPa, and 4 ml/min with the highest lipid yield of 46.74 wt%; above-average findings were reported. Furthermore, the pretreatment process involved significant effects such as crumpled and exhaustive structure, facilitating the efficient extraction of total lipids from the microalgae matrix. This study investigated the microstructure of microalgae biomatrix before and after extraction using scanning electron microscopy (SEM) and thermogravimetric analysis (TGA). Fourier-transform infrared spectroscopy (FTIR) was utilized to assess the potential of the extracted material as a precursor for biodegradable plastic production, with a focus on reduced heavy metal content through inductively coupled plasma-optical emission spectrometry (ICP-OES) analysis. The lipid extracted from Chlorella Vulgaris sp. microalgae was analysed using gas chromatography-mass spectrometry (GC-MS), identifying key constituents, including oleic acid (C18H34O2), n-Hexadecanoic acid (C16H32O2), and octadecanoic acid (C18H36O2), essential for polyhydroxyalkanoate (PHA) formation.

Aqueous enzymatic extraction of macauba (Acrocomia aculeata) pulp oil: A green and sustainable approach for high-quality oil production.

Aqueous enzymatic extraction (AEE) of macauba pulp oil (MPO) was performed in this study with five commercial enzymatic pools. The chemical, nutritional, and thermal properties of the oils with high oil efficiency by AEE were evaluated and compared with mechanical pressing (MP) and organic solvent extraction (SE). Among the AEE processes, the pectinase pool (at pH 5.5 and 50 °C) exhibited the highest process efficiency (88.6 %). The oils presented low acidity values (0.4-3.1 %) and low molar absorptivities, indicating minimal oil degradation. Bioactive compounds, such as carotenoids, were found in MPO. The iodine index and the fatty acid profile of the oils revealed a high content of unsaturated fatty acids, particularly oleic and linoleic acids, with excellent nutritional scores, as evidenced by anti-atherogenicity and anti-thrombogenicity indices. These findings emphasized that AEE is an eco-friendly approach for extracting high-quality MPO with beneficial health compounds for food products.

Deep eutectic solvents for the extraction of polyphenols from food plants.

Deep Eutectic Solvents (DESs) offer a promising, sustainable alternative for extracting polyphenols from food plants, known for their health benefits. Traditional extraction methods are often costly and involve toxic solvents. This review discusses the basic concepts, preparation techniques, and factors influencing the effective and safe use of DESs in polyphenol extraction. DESs' adaptability allows integration with other green extraction technologies, such as microwave- and ultrasound-assisted extractions, enhancing their efficiency. This adaptability demonstrates the potential of DESs in the sustainable extraction of bioactive compounds. Current research indicates that DESs could play a significant role in the sustainable procurement of these compounds, marking an important advancement in food science research and development. The review underscores DESs as a realistic, eco-friendly alternative in the realm of natural extraction technologies, offering a significant contribution to sustainable practices in food science.

Characterizing date seed polysaccharides: A comprehensive study on extraction, biological activities, prebiotic potential, gut microbiota modulation, and rheology using microwave-assisted deep eutectic solvent.

This study investigated the biological activities, prebiotic potentials, modulating gut microbiota, and rheological properties of polysaccharides derived from date seeds via microwave-assisted deep eutectic solvent systems. Averaged molecular weight (246.5 kDa) and a monosaccharide profile (galacturonic acid: glucose: mannose: fructose: galactose), classifying MPS as a heteropolysaccharide. MPS, at concentrations of 125-1000 µg/mL, demonstrates increasing free radical scavenging activities (DPPH, ABTS, MC, SOD, SORS, and LO), potent antioxidant potential (FRAP: 51.2-538.3 µg/mL; TAC: 28.3-683.4 µg/mL; RP: 18.5-171.2 µg/mL), and dose-dependent antimicrobial activity against common foodborne pathogens. Partially-purified MPS exhibits inhibition against α-glucosidase (79.6 %), α-amylase (85.1 %), and ACE (68.4 %), along with 80 % and 46 % inhibition against Caco-2 and MCF-7 cancer cells, respectively. Results indicate that MPS fosters the growth of beneficial fecal microbiota, including Proteobacteria, Firmicutes, and Actinobacteria, supporting microbes responsible for major SCFAs (acetic, propionic, and butyric acids) production, such as Ruminococcus and Blautia.

Resource-Efficient Use of Residues from Medicinal and Aromatic Plants for Production of Secondary Plant Metabolites.

Although people's interest in green and healthy plant-based products and natural active ingredients in the cosmetic, pharmaceutical, and food industries is steadily increasing, medicinal and aromatic plants (MAPs) represent a niche crop type.It is possible to increase cultivation and sales of MAPs, by utilizing plant components that are usually discarded. This chapter provides an overview of studies concerning material flows and methods used for sustainable production of valuable metabolites from MAPs between 2018 and 2023. Additionally, it describes new developments and strategies for extraction and isolation, as well as innovative applications. In order to use these valuable resources almost completely, a systematic recycling of the plant material is recommended. This would be a profitable way to increase sustainability in the cultivation and usage of MAPs and provide new opportunities for extraction in plant science.

Comparison of green solvents for the revalorization of orange by-products: Carotenoid extraction and in vitro antioxidant activity.

Orange peels contain a considerable number of bioactive compounds such as carotenoids, that can be used as ingredients in high-value products. The aim of this study was to compare orange peel extracts obtained with different green solvents (vegetable oils, fatty acids, and deep eutectic solvents (DES)). In addition, the chemical characterization of a new hydrophobic DES formed by octanoic acid and l-proline (C8:Pro) was performed. The extracts were compared in terms of carotenoid extraction, antioxidant activity by three methods, color, and environmental impact. The results confirmed that the mixture of C8:Pro is a DES and showed the highest carotenoid extraction (46.01 µg/g) compared to hexane (39.28 µg/g). The antioxidant activity was also the highest in C8:Pro (2438.8 µM TE/mL). Finally, two assessment models were used to evaluate the greenness and sustainability of the proposed extractions. These results demonstrated the potential use of orange peels in the circular economy and industry.

Evaluating the Antioxidant and Antidiabetic Properties of Medicago sativa and Solidago virgaurea Polyphenolic-Rich Extracts.

The present study evaluated the antioxidant and antidiabetic properties of Medicago sativa and Solidago virgaurea extracts enriched in polyphenolic compounds. The extracts were obtained by accelerated solvent extraction (ASE) and laser irradiation. Then, microfiltration was used for purification, followed by nanofiltration used to concentrate the two extracts. The obtained extracts were analyzed to determine their antioxidant activity using DPPH radical scavenging and reducing power methods. The antidiabetic properties have been investigated in vitro on a murine insulinoma cell line (ß-TC-6) by the inhibition of α-amylase and α-glucosidase. M. sativa obtained by laser irradiation and concentrated by nanofiltration showed the highest DPPH• scavenging (EC50 = 105.2 ± 1.1 µg/mL) and reducing power activities (EC50 = 40.98 ± 0.2 µg/mL). M. sativa extracts had higher inhibition on α-amylase (IC50 = 23.9 ± 1.2 µg/mL for concentrated extract obtained after ASE, and 26.8 ± 1.1), while S. virgaurea had the highest α-glucosidase inhibition (9.3 ± 0.9 µg/mL for concentrated extract obtained after ASE, and 8.6 ± 0.7 µg/mL for concentrated extract obtained after laser extraction). The obtained results after evaluating in vitro the antidiabetic activity showed that the treatment with M. sativa and S. virgaurea polyphenolic-rich extracts stimulated the insulin secretion of ß-TC-6 cells, both under normal conditions and under hyperglycemic conditions as well. This paper argues that M. sativa and S. virgaurea polyphenolic-rich extracts could be excellent natural sources with promising antidiabetic potential.

Ultrasound-assisted and subcritical water extraction techniques for maximal recovery of phenolic compounds from raw ginger herbal dust toward in vitro biological activity investigation.

This study examined the impact of two green extraction techniques in order to maximize the usage and recovery of phenolic compounds from the by-product of the filter tea industry, the so-called ginger herbal dust. The main phenolic compounds extraction was performed by ultrasound-assisted extraction (UAE) with the sonication amplitude ranging from 20% to 100%, and the subcritical water extraction (SWE), with the temperature ranging from 120 °C to 220 °C. All obtained extracts were characterized in terms of extraction yield, total phenolic content (TPC), and 6-ginerol, 6-shogaol, and 8-ginerol contents using RP-HPLC-DAD. Based on the results, we selected the extract obtained from raw ginger herbal dust using a sonication amplitude of 100% for further biological investigation of the cytotoxic effect on short- and long-term cell viability on liver and pancreatic cancer cells. This extract contained high TPC concentration, and 6-gingerol (44.57 mg/gDE), 8-gingerol (8.62 mg/gDE), and 6-shogaol (6.92 mg/gDE).

Design of Experiments for Optimizing Ultrasound-Assisted Extraction of Bioactive Compounds from Plant-Based Sources.

Plant-based materials are an important source of bioactive compounds (BC) with interesting industrial applications. Therefore, adequate experimental strategies for maximizing their recovery yield are required. Among all procedures for extracting BC (maceration, Soxhlet, hydro-distillation, pulsed-electric field, enzyme, microwave, high hydrostatic pressure, and supercritical fluids), the ultrasound-assisted extraction (UAE) highlighted as an advanced, cost-efficient, eco-friendly, and sustainable alternative for recovering BC (polyphenols, flavonoids, anthocyanins, and carotenoids) from plant sources with higher yields. However, the UAE efficiency is influenced by several factors, including operational variables and extraction process (frequency, amplitude, ultrasonic power, pulse cycle, type of solvent, extraction time, solvent-to-solid ratio, pH, particle size, and temperature) that exert an impact on the molecular structures of targeted molecules, leading to variations in their biological properties. In this context, a diverse design of experiments (DOEs), including full or fractional factorial, Plackett-Burman, Box-Behnken, Central composite, Taguchi, Mixture, D-optimal, and Doehlert have been investigated alone and in combination to optimize the UAE of BC from plant-based materials, using the response surface methodology and mathematical models in a simple or multi-factorial/multi-response approach. The present review summarizes the advantages and limitations of the most common DOEs investigated to optimize the UAE of bioactive compounds from plant-based materials.

Recent Progress on Green New Phase Extraction and Preparation of Polyphenols in Edible Oil.

With the proposal of replacing toxic solvents with non-toxic solvents in the concept of green chemistry, the development and utilization of new green extraction techniques have become a research hotspot. Phenolic compounds in edible oils have good antioxidant activity, but due to their low content and complex matrix, it is difficult to achieve a high extraction rate in a green and efficient way. This paper reviews the current research status of novel extraction materials in solid-phase extraction, including carbon nanotubes, graphene and metal-organic frameworks, as well as the application of green chemical materials in liquid-phase extraction, including deep eutectic solvents, ionic liquids, supercritical fluids and supramolecular solvents. The aim is to provide a more specific reference for realizing the green and efficient extraction of polyphenolic compounds from edible oils, as well as another possibility for the future research trend of green extraction technology.

Polyphenol Extraction from Food (by) Products by Pulsed Electric Field: A Review.

Nowadays, more and more researchers engage in studies regarding the extraction of bioactive compounds from natural sources. To this end, plenty of studies have been published on this topic, with the interest in the field growing exponentially. One major aim of such studies is to maximize the extraction yield and, simultaneously, to use procedures that adhere to the principles of green chemistry, as much as possible. It was not until recently that pulsed electric field (PEF) technology has been put to good use to achieve this goal. This new technique exhibits many advantages, compared to other techniques, and they have successfully been reaped for the production of extracts with enhanced concentrations in bioactive compounds. In this advancing field of research, a good understanding of the existing literature is mandatory to develop more advanced concepts in the future. The aim of this review is to provide a thorough discussion of the most important applications of PEF for the enhancement of polyphenols extraction from fresh food products and by-products, as well as to discuss the current limitations and the prospects of the field.

Green Extraction and Preliminary Biological Activity of Hydrolyzed Collagen Peptides (HCPs) Obtained from Whole Undersized Unwanted Catches (Mugil cephalus L.).

Considering the global increase in fish consumption, the growing side-streams coming from the fish supply chain (e.g., skin, fins, tail, heads…), also including undersized or "unwanted catches", have been recently proposed as source of high-value bioactive compounds (e.g., peptides and fatty acids). In this case study, hydrolyzed collagen peptides (HCPs) were extracted from different parts of Mugil cephalus L. using environmentally friendly techniques such as ultrasounds and enzymatic treatments. Both a mixed biomass derived from the skin, fins, and tail, and a whole fish, were considered as starting biomass, simulating the unsorted processing side-streams and an undersized/unwanted catch, respectively. The extracted HCPs were purified in fractions (<3 KDa and >3 KDa) whose yields (about 5% and 0.04-0.3%, respectively) demonstrated the efficiency of the hydrolysis process. The extraction protocol proposed allowed us to also isolate the intermediate products, namely the lipids (about 8-10%) and the non-collagenous proteins (NCs, 16-23%), whose exploitation could be considered. Each sample was characterized using Sircol, UltraViolet-Spectra, and hydroxyproline assay, and the viability of their collagen fractions was tested on human endothelial cells. Significant effects were obtained at a fraction of <3 KDa, in particular at a concentration of 0.13 µg/mL. The T-scratch test was also performed, with positive results in all fractions tested.