A method for obtaining collagen peptides and valuable intermediate products with green production technology

Abstract

The invention relates to a method for obtaining functional and high value collagen peptides from collagen sources using supercritical fluid technology. This method also enables the production of other valuable intermediates. This method has green technological features and offers an environmentally friendly approach by minimizing the use of chemical solvents. It enables the production of high purity and valuable products in a short period of time and provides an efficient production process at low costs.

Description

[0001] A METHOD FOR OBTAINING COLLAGEN PEPTIDES AND VALUABLE INTERMEDIATE PRODUCTS WITH GREEN PRODUCTION TECHNOLOGY TECHNICAL FIELD

[0002] The invention relates to a method for obtaining functional and high-value collagen peptides from collagen sources using supercritical fluid technology. This method also enables the production of other valuable intermediate products. With its green technological features, this method offers an environmentally friendly approach by minimizing the use of chemical solvents. It enables the rapid production of high-purity, valuable products while providing an efficient production process at low costs.

[0003] PRIOR ART

[0004] Collagen is a structural protein that provides strength and connectivity between tissues, constituting approximately 30% of the human body and 75% of the skin. However, collagen levels in the body decrease with age. Collagen deficiency can lead not only to aesthetic problems such as wrinkles and signs of aging but also to serious health issues such as joint and bone problems and osteoporosis. Therefore, individuals need to regularly supplement their bodies with collagen. Especially for individuals over the age of 20, the recommended daily collagen intake is 10 grams.

[0005] In current practice, collagen is generally derived from bovine, porcine, fish, and chicken sources. In Turkiye, various methods are applied in collagen production, with bovine byproducts being the most commonly used source. However, the production processes used in the existing techniques are often inadequate due to their high costs, intensive labor requirements, and time-consuming steps. The main reason for this is the need for long processing times and complex procedures to obtain collagen from animal waste and / or byproducts.

[0006] In order to offer collagen and collagen-containing products to the market at affordable prices, there is a need for production methods that reduce labor requirements, lower process costs, and enable operations to be completed in a shorter time. Such improvements will both enhance production efficiency and allow the products to become accessible to a broader range of consumers.On the other hand, in terms of technology, high-cost acidic and / or enzymatic methods are generally used for collagen extraction. However, traditional methods such as acidic, basic, or enzymatic extraction and hydrolysis processes stand out due to their complex structures, high energy and resource consumption requirements, long process times, and low efficiency. These factors necessitate the development of more efficient, economical, and environmentally friendly alternative methods for collagen production.

[0007] Since the separation and purification of collagen sources involves multiple steps, the process can take a long time to complete. This reduces production efficiency and increases susceptibility to disruptions in the process.

[0008] The use of high temperatures and harsh chemicals in traditional methods can negatively affect the purity and quality of the final product. In particular, these methods may degrade sensitive protein and lipid structures, thereby reducing the biological activity of the product.

[0009] In line with the objectives of achieving energy savings, increasing capacity utilization, and reducing costs in extraction processes, ongoing efforts aim to enhance processing speed and efficiency without compromising quality. In this context, the development of new techniques and the use of alternative solvents have come to the forefront. Supercritical fluid extraction, discovered approximately 20 years ago, emerged as a result of these efforts and has found widespread use in various industries, including food, cosmetics, and pharmaceuticals. Supercritical fluid extraction provides commercially satisfactory results in the production of high-quality essential oils with lower monoterpene content compared to those obtained through conventional methods.

[0010] The critical temperature and pressure of each substance define the highest conditions under which its gas and liquid phases can coexist. Above these critical values, the substance enters a distinct phase known as a supercritical fluid. Supercritical fluids possess physical and transport properties that are intermediate between those of gases and liquids, while their solvating power is comparable to that of liquids. Their density varies depending on parameters such as pressure and temperature, which enables high speed and efficiency in extraction processes.

[0011] The supercritical extraction process is based on dissolving a substance in a supercritical fluid, then reducing the pressure to separate the product. When the pressure is reduced, the solubility of the dissolved substance in the supercritical phase decreases and separation occurs. Carbon dioxide is commonly used as the supercritical fluid. The use of CO2offers amore environmentally friendly approach compared to organic solvents while increasing process speed. Supercritical carbon dioxide has additional advantages such as reducing microbial load and inactivating unwanted enzymes. For example, studies conducted on orange juice have shown that carbon dioxide improves product quality by rendering the pectinesterase enzyme inactive. Additionally, supercritical carbon dioxide has the ability to significantly eliminate pathogens such as Listeria and Salmonella. This property offers great potential in the processing of collagen sources for reducing microbial load and eliminating unwanted odors.

[0012] Supercritical fluid technology is employed not only in microbial load reduction and extraction processes but also in applications such as particle size control and enhancement of biological activity. Various crystallization techniques utilizing supercritical fluids allow for the production of high-purity products with narrow particle size distributions, without the need for organic solvents. For instance, rapid expansion of a supercritical solution leads to high supersaturation levels, resulting in the formation of small and homogeneous particles. These methods have yielded promising results in the production of protein and peptide powders and have demonstrated superior performance compared to conventional techniques, particularly in terms of maintaining biological activity. Such innovative technologies provide a significant competitive advantage across multiple industries by improving both process efficiency and product quality.

[0013] Solvents and chemical agents such as acidic, basic, or EDTA-based solutions are commonly employed to separate oil and non-collagenous components from collagen sources. However, such practices may cause environmental harm during production and increase the risk of chemical residues remaining in the final product. With growing global awareness of environmental challenges, the concept of green technology has gained significant importance in recent years. In this context, it is essential to reduce the generation and use of hazardous substances, minimize energy consumption, and promote the utilization of renewable resources. The disposal of chemical solvents and their associated waste poses environmental risks, and the carbon footprint generated by conventional production methods is often considerable. These factors create a framework that is inconsistent with established sustainability objectives.

[0014] In conclusion, the development of innovative methods is of great importance to address or minimize the problems associated with current techniques. Such methods should aim to reduce environmental impact while ensuring high purity and quality, lowering microbial load, and minimizing undesirable odor components. Furthermore, they should enable productionwithin a shorter time frame, at lower cost, and based on a zero-waste principle through the use of inexpensive, recyclable, and environmentally friendly green solvents. This approach is intended to provide effective and sustainable solutions, particularly for the sensitive requirements of industries such as food, healthcare, and cosmetics.

[0015] BRIEF DESCRIPTION OF THE INVENTION

[0016] The present invention relates to a green technological method for obtaining collagen peptides and other valuable intermediate products from collagen sources, with the aim of eliminating the disadvantages described above and introducing new advantages to the relevant technical field.

[0017] The object of the present invention is to provide a more effective and efficient production method by eliminating the lengthy, costly, and complex process steps involved in current collagen peptide production techniques. This approach optimizes the production process, reduces operational costs, and minimizes labor requirements.

[0018] Another object of the present invention is to provide a collagen peptide production method that minimizes the use of toxic chemicals and reduces environmental impact. In this way, a method in line with the concept of green technology can be developed through the implementation of sustainable production processes.

[0019] Another object of the present invention is to provide a method for obtaining collagen peptides with high quality and purity. Thus, the production of collagen peptides that comply with industrial standards and enhance consumer confidence becomes achievable.

[0020] Another object of the present invention is to establish a production method that allows for the recovery of collagen peptides as the main product, along with other valuable intermediate products from the source materials, at reasonable costs and in alignment with the zero-waste objective.

[0021] DETAILED DESCRIPTION OF THE INVENTION

[0022] The following detailed description relates to a method for obtaining collagen peptides from collagen sources and is provided by way of example only, with the intention of facilitating understanding of the invention, and not for the purpose of limiting its scope in any way.The term “animal sources” as used in the present invention refers to any animal materials suitable for collagen production, regardless of their collagen content, and denotes the raw material in its initial, unprocessed form. The term “collagen source” refers to animal materials that have undergone preliminary processing.

[0023] Preferably, fish, chickens, cattle, small ruminants, pigs, or eggs can be used as animal sources. However, the method described in this invention is not limited to these sources and can be applied to all animal sources for collagen extraction.

[0024] Another important aspect of the present invention is that it offers technical advantages such as cost reduction and waste recycling by enabling the use of animal parts that are typically considered waste or by-products as collagen sources. For example, animal waste materials such as fish bones, scales, and skin may be regarded as suitable sources for collagen extraction within the context of this invention. For the sake of simplicity, the term “source” may sometimes be used in this specification to refer collectively to both animal sources and animal waste materials used in collagen production. When the term “animal waste sources” is employed, a person skilled in the art will readily understand that this refers to the valorization of animal waste for collagen production purposes.

[0025] In this invention, collagen is addressed as a structural protein found in the human body. Although 28 different types of collagens have been identified in the literature, the invention is not limited to the production or use of a specific collagen type. The type of collagen obtained may vary depending on the raw material used. The method described herein is applicable to the extraction and processing of any desired collagen type. Accordingly, the invention provides a flexible and versatile approach, enabling broad application across various collagen types without restriction to a particular form.

[0026] In this invention, the term “valuable intermediate products” refers to products with commercial value obtained during the processing of collagen sources, such as collagen peptides, which are the main product, as well as fish oil, fish protein, fish meal or feed, chicken oil, chicken protein, and gelatins, as preferred. These products can be used in various sectors such as food, health, cosmetics, and agriculture, enhancing the economic efficiency of the processing process and contributing to an environmentally friendly production model by utilizing waste.

[0027] The invention relates to a method for obtaining collagen peptides, which comprises the following steps:Cleaning and sorting of raw materials prior to their evaluation as a collagen source:

[0028] In this process step, raw materials that can be considered as collagen sources are cleaned by removing blood, internal organs, and other foreign substances. The cleaning process involves washing the raw materials, preferably with water, and separating them from organic residues. At this stage, foreign substances that could reduce quality are separated, and the raw materials are made suitable for the production process.

[0029] In the preferred application, fish or chicken is used as the collagen source. For fish waste in particular, collagen-rich parts such as skin, scales, bones, and spines are carefully separated from blood, internal organs, and other organic residues. Hygiene is maintained during the washing process, and internal organs or unwanted elements are carefully examined to ensure they have not been mixed in. If any internal organ residues are detected, they are physically separated to ensure purity. This cleaning process not only enhances the purity of the raw material but also prevents biological contaminants that could negatively affect the quality of collagen and other intermediate products. Additionally, this stage improves the efficiency of subsequent processes in the production line and prevents unwanted contamination.

[0030] Breakdown of cleaned collagen sources and size reduction to target

[0031] The raw material fragmentation process step is a process that breaks down collagen sources into smaller sizes. This process step plays a critical role in the processing of raw materials with large and complex structures, such as mixed fish waste.

[0032] The grinding process increases the surface area of the raw material, thereby enhancing the efficiency of subsequent processes such as oil and moisture separation, protein separation, and extraction. Particulated raw materials, due to their homogeneous structure, enable chemical or physical processes applied throughout the process to penetrate all parts of the raw material evenly. This results in a production process that enhances process efficiency, leading to higher quality and productivity.

[0033] The equipment employed during this stage typically includes shredders or grinders. Hard and bulky materials such as fish or chicken skin, bones, and spines are reduced to smaller fragments using these machines.Ancak, However, this process is not usually necessary for small and light components such as fish scales.Broken-down raw materials become more suitable for subsequent processing steps. In particular, the solvent used in supercritical fluid systems can penetrate deeper into the inner layers of the raw material, making it easier to separate oil and moisture and thereby enhancing the efficiency of these separation processes. Additionally, after the fragmenting process, the raw material becomes easier to transport and can be processed with less energy in the production line. As a result of this process, the cleaned materials can be reduced to a size ranging from 1 to 200 mm.

[0034] Removal of oil and moisture from collagen sources reduced to target dimensions in a supercritical fluid system

[0035] The separation of oil and moisture from collagen sources in a supercritical fluid system is a critical process step for the processing of collagen sources, which is the subject of the invention. First, the collagen source, which has been broken down and prepared for processing, is placed into the supercritical fluid system. This system operates using a supercritical fluid that acts as a solvent with high efficiency, possessing both gas and liquid properties. The process is carried out under low temperature and pressure conditions in an environmentally friendly manner. Since no solvent is used, environmental impacts are minimized. Additionally, the separation of oil and moisture eliminates the need for storing collagen sources in a cold storage facility, allowing them to be preserved at room temperature. This reduces energy and logistics costs, while also increasing process efficiency by saving storage space through the elimination of drying and grinding of the raw material.

[0036] In a preferred embodiment, at least one of carbon dioxide, water, and ethanol is used as the solvent in the supercritical fluid system.

[0037] One of the most preferred applications is the use of carbon dioxide (CO2) as a solvent in supercritical fluid systems. CO2's nonpolar structure effectively separates oil-soluble compounds, enabling the production of by-products with high purity and quality. Additionally, the high moisture absorption capacity of the supercritical fluid system facilitates the drying of collagen sources while enabling the processing of valuable intermediate products in a purer form. The low temperature and pressure conditions used during this process preserve the natural properties of oils and proteins, thereby enhancing the biological activity of the products. The non-toxic nature of supercritical CO2and its conversion back into a gas phase at the end of the process ensures a pure production without leaving chemical residues in the products. Furthermore, the recovery and reuse of a significant portion of CO2within thesystem reduces costs and provides an environmentally friendly production process. This method not only enhances product quality but also supports a sustainable production approach.

[0038] In this process step, the ambient temperature is maintained at a value between 31 and 80 °C to separate oil and moisture from collagen sources. During static holding and dynamic flow, the ambient pressure is maintained between 70 and 400 bar. Depending on these conditions, the static holding and dynamic flow process duration ranges from 15 to 250 minutes, optimized according to the type and composition of the collagen source to achieve the most efficient results. Thanks to these process parameters, the moisture level in collagen sources is preferably reduced below 15%, providing an appropriate structure for subsequent process steps. Additionally, during this process, pressure changes are not sudden but are implemented through a gradual decrease, ensuring the process progresses in a controlled manner.

[0039] The flow rate in a supercritical fluid system using CO2as a solvent is in the range of 0.1 to 6 kg / min.

[0040] Similarly, the process of separating oil from collagen sources is also carried out using these parameters. This process is applied by utilizing the high solubility of the supercritical fluid system. The use of low temperatures helps preserve the natural structure of the oil, and since no solvents are required, no chemical residues are present in the final product. As a result of the processes carried out using this method, the collagen source obtained achieves oil values preferably below 5%.

[0041] Thanks to the process parameters defined at this stage, it becomes possible to obtain not only collagen peptides but also other valuable intermediate products. For instance, when fish-based collagen sources are used, fish oil can be recovered; similarly, when chickenbased collagen sources are utilized, chicken oil can be obtained.

[0042] - Separation of non-collagen proteins

[0043] This process step is performed by washing the defatted and dehydrated collagen sources with weakly acidic or basic solutions at low concentrations, or with cold water, at a temperature between 0°C and 25°C. Since collagen is a protein that dissolves at high temperatures, it remains insoluble and preserved within the raw material at low temperatures. In contrast, non-collagen proteins — some of which may be consideredvaluable intermediate products — dissolve in the solution and are separated. This method not only increases the purity of collagen but also improves process efficiency by facilitating downstream processing steps.

[0044] During the separation of non-collagen proteins, valuable intermediate products such as fish or chicken proteins can be obtained. In a preferred embodiment, this process is carried out by washing the raw material — previously dehydrated to reduce its moisture content — with cold water at a temperature between 0°C and 25°C. As collagen is a protein that dissolves at high temperatures, it remains insoluble within this temperature range, whereas noncollagen proteins dissolve and are separated from the raw material. The extracted noncollagen proteins can be further purified and utilized in sectors such as dietary supplements or animal feed. This process not only increases the purity of the collagen fraction but also creates economic value by enabling the commercial use of the separated proteins. The use of cold water in this step provides a simple, low-cost, and environmentally friendly approach, making the process both efficient and sustainable.

[0045] The fact that this process step is carried out with water makes the method low cost and simple. It offers both environmental and economic advantages.

[0046] Performing extraction and partial demineralization processes

[0047] In the method of the present invention, the extraction and partial demineralization steps are carried out within a supercritical fluid system.

[0048] In the mentioned supercritical process step, at least one solvent selected from water, carbon dioxide, deep eutectic solvents, or weak acids or bases is used.

[0049] In a most preferred embodiment, water and deep eutectic solvents suitable for collagen peptides are used as solvents for supercritical extraction processes. Additionally, carbon dioxide (CO2) may be incorporated into the supercritical fluid system as a solvent, if preferred.

[0050] Deep eutectic solvents are defined as mixtures of two or more compounds that, when combined, form a eutectic system with a melting point lower than that of each individual component, as illustrated in the phase diagram. The formulation of deep eutectic solvents usually consists of binary mixtures. In these mixtures, one of the components acts as hydrogen bond acceptor (abbreviated as HBA) and the other as hydrogen bond donor(abbreviated as HBD). In solvent formation, hydrogen bonds occur between compounds instead of covalent bonds. The ratio between the hydrogen bond acceptor and the hydrogen bond donor that make up the eutectic varies depending on the mutual hydrogen bonding ability of these components.

[0051] Deep eutectic solvents facilitate the extraction process by weakening the molecular bonds within the collagen source. These solvents also regulate the pH level, thereby enhancing the efficiency of the extraction process.

[0052] In this process step, the collagen sources are preferably first placed in distilled water. The weight ratio of collagen source to distilled water is preferably in the range of 1 :5 to 1 :30. Subsequently, a deep eutectic solvent is added to the processing environment. The amount of the deep eutectic solvent is determined to be between 0.1% and 35% by weight of the resulting solution. These operations lower the pH value within the supercritical fluid system and allow the deep eutectic solvent to interact with the hydrogen bonds between collagen molecules, thereby facilitating the loosening of the collagen structure and making it more suitable for extraction.

[0053] In the invention, the deep eutectic solution used in the extraction processes comprises at least one of choline chloride, choline acetate, choline bromide, citric acid, betaine, or glucose as the hydrogen bond acceptor (HBA) component.

[0054] In the invention, the deep eutectic solution used for extraction processes comprises at least one of lactic acid, acetic acid, oxalic acid, glycolic acid, glycerol, citric acid, or malic acid as HBD component.

[0055] The deep eutectic solution according to the invention contains HBA:HBD in a weight ratio of 1 :0.33 to 1 :3.

[0056] In the extraction step, suitable production parameters are determined within the supercritical fluid system. The temperature for this step is in the range of 40 to 100°C. The flow rate of the solvent used ranges from 0.1 to 25 liters per minute. In a preferred embodiment, the flow rate of the solvent is regulated and fed into the system using a pump. In another preferred embodiment, the system pressure is between 75 and 380 bar, and most preferably between 180 and 350 bar. Depending on the selected parameters, the static holding time during this step ranges from 15 to 200 minutes, while the dynamic flow time is between 30 and 200 minutes. The pH of the medium in which the process is carried out is between 4 and 8.As it is known, at supercritical fluid system conditions, there are changes in diffusion and viscosity, which are the source of the mass transfer ability of the dense gas. Under these conditions, although the gas is dense like a liquid, it is sociable like a gas and its viscosity is low like gas viscosity. These properties and low surface tension increase its ability to enter pores and dissolve substances. The ambient conditions of the invention are optimized to accelerate the extraction process.

[0057] After the extraction process is completed, during the transfer of the product from the extractor to the separators and / or discharge from the separator, a sudden drop to a pressure much lower than the applied pressure is applied, preferably within a short time. This pressure change also performs a partial demineralization process. In this process, thanks to the selectivity of the supercritical fluid system, the protein structures in the collagen sources are removed with high efficiency, while the passage of minerals and insoluble substances is limited.

[0058] Partial demineralization via Rapid Expansion of Supercritical Solvents (RESS) is achieved by reducing the extraction pressure to a range between 0 and 50 bar. This pressure drop may be applied either gradually and / or instantaneously over a duration of 0.1 to 20 minutes. As previously noted, the extraction step is carried out under pressures ranging from 75 to 380 bar, which are subsequently lowered in a controlled manner to facilitate the transfer of the product into hydrolysis tanks for further processing. These controlled pressure changes promote rapid and efficient fractionation of minerals and insoluble compounds, while simultaneously contributing to the reduction of particle and molecular sizes. As a result, the subsequent hydrolysis processes of collagen sources are rendered more efficient and effective.

[0059] After this process step, liquid gelatin solution with lower molecular weight and high viscosity values is obtained from collagen molecules with a molecular weight of approximately 300 kDa by applying supercritical fluid technology.

[0060] If the gelatin solution obtained at this stage is not subjected to the subsequent hydrolysis process step but instead directed to alternative processing steps, gelatin can be obtained depending on the source material used. Gelatin derived from animal sources is widely utilized across various industries including food, pharmaceuticals, cosmetics, and industrial applications. In the food industry, it serves as a thickening, stabilizing, and gelling agent in products such as confectionery, yogurt, ice cream, jelly, meat products, and functional foods. In the pharmaceutical industry, gelatin is employed in the production of capsules, as a tabletbinder, and as a component in wound healing formulations. In cosmetic applications, it is incorporated into skincare products, hair gels, and anti-aging creams for its moisturizing and structural support properties. Furthermore, gelatin finds extensive use in technical fields such as photographic film coatings, as an adhesive in paper manufacturing, and as a finishing agent in the textile industry.

[0061] This process step provides an environmentally friendly approach by minimizing solvent usage. Simultaneously, it enables the rapid obtainment of high-purity and high-quality products. The integration of deep eutectic solvents with the supercritical extraction system enhances extraction efficiency, conserves energy, and reduces processing costs. As such, the process represents an eco-friendly and cost-effective production model.

[0062] In a preferred embodiment of the invention, the extraction process is carried out using deep eutectic solvents, and this process is optimized by supercritical fluid and / or ultrasound applications. The ratio of deep eutectic solvent used during extraction is determined between 1 :5 and 1 :20 by weight depending on the collagen source and the process temperature is preferably kept between 25 °C and 95 °C. Supercritical fluid (preferably CO2) extraction can be applied during or simultaneously with this process, and ultrasound waves in the frequency range of 15 kHz to 45 kHz are applied to the extraction medium to improve solubility and selectivity. The duration of ultrasound application is preferably between 5 minutes and 3 hours and ensures the breakdown of cellular structures, solvent penetration and efficient extraction of target compounds.

[0063] This integrated extraction method increases the efficiency of extraction with deep eutectic solvents and increases the yield of collagen. It also reduces the need for solvents, shortens the process time and reduces production costs. In this respect, ultrasound technology is an important environmentally friendly supportive application in terms of sustainable green production.

[0064] Performing the hydrolysis process step

[0065] Gelatin obtained from the collagen source in the previous process steps is subjected to enzymatic hydrolysis to produce collagen peptides. This is a controlled biochemical process in which one or more specific enzymes cleave the collagen chains into low molecular weight peptides. Enzymatic hydrolysis improves the bioavailability and functional properties of the final product while ensuring high purity and process efficiency.One or more enzymes are used to carry out the hydrolysis process step. As enzymes, in the present invention, at least one of the group of endo or exo proteases, alkalases, pepsin, trypsin, papain is preferred.

[0066] After the extraction process is completed, the collagen sources are taken and placed in the enzyme solution. In this process, the collagen sources are hydrolyzed under controlled conditions at a temperature range of 30°C to 80°C, a pH value between 4 and 8.5 and a duration of 1 to 6 hours. This process allows the enzymes to break down the collagen chains into smaller peptides, resulting in collagen peptides with high purity and high bioavailability.

[0067] After hydrolysis is completed, the activity of the enzymes is terminated and the reaction is stopped. In order to inactivate the enzyme and inactivate possible microorganisms, the mixture is passed through a sterilizer at a temperature range of 80°C to 120°C to perform both enzyme deactivation and sterilization.

[0068] In a preferred embodiment, the amount of enzyme is in the range of 0.1% to 5% by weight in the collagen source.

[0069] Enzymatic hydrolysis saves energy as it takes place at low temperatures, preserves delicate protein structures and increases the bioavailability of the product. This method is widely used, especially in the production of delicate and high-quality collagen peptides.

[0070] As a result of hydrolysis, the collagen chains present in the raw material are cleaved to yield bioactive collagen peptides with a molecular weight of less than 5000 Daltons, and preferably less than 2000 Daltons. These peptides consist of short amino acid chains and exhibit high bioaccessibility, bioavailability, and digestibility in the gastrointestinal tract.

[0071] Performing filtration processes to collagen sources with hydrolysis processes

[0072] The filtration process is preferably performed using one or more of microfiltration, ultrafiltration, or membrane filtration methods. The filtration process also acts as a demineralization process, resulting in a more pure, homogeneous and highly bioaccessible form of collagen peptides. At the same time, filtration makes the raw material suitable for subsequent drying and packaging steps.

[0073] Collagen hydrolysate solution, whose molecular size is reduced by the hydrolysis process, is preferably ultrafiltration, microfiltration, membrane filtration method filtration processes areperformed. For this process, processes are preferably carried out with a pressure in the range of 1 to 15 bar.

[0074] Performing purification processes

[0075] Depending on the type, the collagen source may contain varying amounts of undesirable odors, minerals and heavy metals. Activated carbon filters and ion exchange systems are used for these processes. In the purification stage, anionic, cationic, deodorizing and / or mixbed resin systems are used to remove anionic and cationic charged minerals, heavy metals and raw material and / or process-induced odor problems from the solution through activated carbon filters and resins. In this way, demineralization process is also performed and the ash content of the final product to be obtained is reduced below 5%. It is purified from heavy metals. The protein purity of the collagen hydrolysate is preferably above 90%. As a result of these processes, it is possible to obtain collagen peptides of 5000 daltons and preferably below 2000 daltons. This is advantageous in the food and cosmetics industry.

[0076] Processes for increasing concentration

[0077] In this process step, the concentration of the liquid solution of the collagen peptides subjected to purification processes is increased and the amount of water is evaporated in evaporators or molecular distillation devices or at least one of the equipment with the same purpose. This process continues until the water content is preferably in the range of 40% to 60% by weight.

[0078] In a preferred embodiment, the liquid obtained during this process step can be recovered. In this way, 80% or 90% of the water used in the system can be recovered.

[0079] Performing drying processes

[0080] The resulting solution of collagen peptides is subjected to drying processes.

[0081] Drying processes include at least one of the following methods: evaporator drying, spray dryer spray drying, freeze drying.

[0082] Drying processes can be carried out by a spray dry method if preferred. The inlet temperature of the mentioned method is in the range of 120 to 190°C and the outlet temperature is in the range of 45 to 110°C.Drying processes can be carried out in a lyophilizer if preferred. The temperature of the mentioned method is a value of -50°C or lower.

[0083] Owing to the method of the present invention, the protein purity of collagen peptides obtained from collagen-containing animal sources such as fresh or saltwater fish, chicken, eggs or from the wastes of these animal sources is between 90% and 98%. In this way, high purity bioactive collagen peptides suitable for use in various technical fields can be obtained. Furthermore, the ash content of the collagen peptides produced by the subject method is less than 5%, preferably less than 1%. This feature contributes to obtaining pure collagen peptides. The moisture content of the collagen peptides produced by the method according to the invention is between 4% and 10%.

[0084] The collagen peptides obtained through the method described in this invention have a molecular weight of less than 5000 Daltons, preferably around 2000 Daltons. This characteristic enables the peptides to exhibit high bioaccessibility and bioavailability.

[0085] In a preferred embodiment of the invention, it has been determined that when fish skins are used as a collagen source, a high proportion of Type I collagen is obtained, whereas Type VI and Type X collagens are present in lower amounts. Similarly, collagen peptides derived from fish waste or intermediate products preferably contain a high level of Type I collagen, along with minor amounts of Types II, IV, V, X, XI, XVIII, XXII, and XXVII collagen. Collagen peptides obtained from chicken waste or intermediate products preferably contain a high proportion of Type II collagen, and lower levels of Type I, Type IX collagen, and elastin. These compositions have been confirmed through LC-QTOF analysis.

[0086] The pH value of collagen peptides obtained by the method of the invention is in the range of 6 to 7. Thanks to these pH values, suitable values can be obtained for use in different technical fields. In addition, in microbiological analyzes, it was determined that the E. coli and Salmonella ratios of the collagen sources obtained were negative in 10 grams and 25 grams, respectively. The total bacterial count was approximately 1000 CFU / g and the yeast to mold ratio was 30 CFU / g or less. These properties make it an ideal product for use in food and cosmetics. In the heavy metal analysis of the obtained collagen peptides, cadmium content was less than or equal to 1 ppm, lead content was less than 1 ppm and mercury content was less than or equal to 0.1 ppm. This increases the safety and purity of collagen peptides.It was determined that the collagen peptides obtained within the scope of the invention showed 10% and preferably higher cell viability in cell viability analysis performed by MTT method in L929 mouse fibroblast cell line. In this way, it is possible to obtain collagen peptides with cell regenerating effect, especially suitable for use in technical fields such as food and cosmetics.

[0087] The scope of protection of the invention is set out in the appended claims and cannot be strictly limited to what is described in this detailed description for the purpose of illustration. It is obvious that a person skilled in the art can come up with similar constructions in the light of the foregoing without departing from the main theme of the invention.

Claims

CLAIMS1. A method for obtaining collagen peptides and valuable intermediate products from freshwater or saltwater fish or chicken sources, or from parts of these animal sources that can be considered as waste, characterized in that it comprises the following process steps:- pre-treatment of animal resources and / or waste animal resources,• sorting and cleaning of animal resources and / or waste animal resources, • obtaining particle collagen sources by applying size reduction processes to the cleaned sources,- degreasing and dehumidification of collagen sources reduced to target particle sizes by a supercritical fluid system, and the obtaining of fish oil or chicken oil from these sources depending on the animal originthe degreasing and dehumidification processes mentioned herein are carried out in the supercritical fluid system at a value in the range of 31 to 80°C and a pressure value in the range of 70 to 400 bar,- separation of non-collagen proteins by washing the collagen sources, which have been degreased and dehumidified, with at least one weakly acidic solution, at least one basic solution, or water at a temperature in the range of 0 to 25 °C, and obtaining fish protein or chicken protein as non-collagen proteins depending on the animal source used in this process step;- applying extraction and partial demineralization processes to collagen sources purified by the separation of non-collagen proteins using a supercritical fluid system,o the collagen sources are first added to water and the mixture is obtained, o adding a deep eutectic solution comprising HBA and HBD components to the obtained mixture, and obtaining a gelatin-containing liquid through supercritical fluid extraction,o the extraction process with the mentioned supercritical fluid system is carried out at a temperature in the range of 40 to 100°C and a pressure in the range of 75 to 400 bar,o partial demineralization of the supercritical fluid system by a gradual and / or sudden reduction of a pressure value in the range of 75 to 400 bar to a value between 0 and 50 bar in the range of 0.1 to 20 minutes, o collection of particles from the mixture remaining as insoluble matter for the production of fish feed, fish meal, chicken feed or chicken meal, depending on the animal source used,- if it is desired to obtain chicken or fish gelatin from the liquid phase, proceed to the next processing step without hydrolysis,If collagen peptides are to be obtained, the gelatin obtained is subjected to hydrolysis by enzymatic method,- inactivation of enzymes and microorganisms by passing the welds obtained from the previous processing steps through the sterilizer,- application of filtration and purification processes,- subjecting the purified collagen peptides or fish or chicken gelatins to concentration increasing and drying processes2. A method according to claim 1 ; characterized in that the collagen sources are reduced into fragments having a size between 1 mm and 200 mm.

3. A method according to any one of the preceding claims; characterized in that at least one of water, carbon dioxide, or an ethanol group solvent is used in the supercritical fluid system during the degreasing and dehumidification process step.

4. A method according to any one of the preceding claims; characterized in that in extraction and partial demineralization process step, the collagen source:water value is in the range of 1 :5 to 1 :25 by weight.

5. A method according to any one of the preceding claims; characterized in that, in the extraction and partial demineralization process step, the deep eutectic solution is added to the collagen source-water mixture in an amount ranging from 1% to 30% by weight.

6. A method according to any one of the preceding claims; characterized in that the deep eutectic solution used in the extraction and partial demineralization processes contains at least one of lactic acid, acetic acid, oxalic acid, oxalic acid, glycolic acid, citric acid, malic acid group to provide HBD property.

7. A method according to any one of the preceding claims; characterized in that the deep eutectic solution used in the extraction and demineralization processes comprises at least one of choline chloride, choline acetate, choline bromide, citric acid, or betaine to provide the HBA property.

8. A method according to any one of the preceding claim; characterized in that the deep eutectic solution contains HBA:HBD in a ratio of 1 :0.33 to 1 :3 by weight.

9. A method according to any one of the preceding claims; characterized in that it contains at least one of endo or exo proteases, alkalases, pepsin, trypsin, papain group as enzymes in hydrolysis processes.

10. A method according to any one of the preceding claims; characterized in that the hydrolysis is carried out at a temperature in the range 30 to 80 °C.11 . A method according to any one of the preceding claims; characterized in that the amount of enzyme in the hydrolysis processes is in the range of 0,1% to 5% by weight, depending on the collagen source.

12. A method according to any one of the preceding claims; characterized in that the hydrolysis processes have a pH value in the range of 4 to 8.

13. A method according to any one of the preceding claims; characterized in that at least one of microfiltration, ultrafiltration, or membrane filtration is applied as said filtration process.

14. A method according to any one of the preceding claims; characterized in that said purification is carried out by at least one of activated carbon, anionic, cationic and mixbed resins.

15. A method according to any one of the preceding claims; characterized in that said concentration increase is carried out by means of an evaporator or molecular distillation device.

16. A method according to any one of the preceding claims, characterized in that after said concentration process step, drying is carried out by lyophilization processes at a temperature of up to -50°C.

17. A method according to any one of the preceding claims, characterized in that, after said concentration process step, a spray drying process step is carried out at an inlet temperature in the range of 120 to 190 °C and an outlet temperature in the range of 45 to 110 °C.