Dry collagen powder with satiating properties, as well as a food product containing collagen powder.

Abstract

Dry collagen powder, characterized by the fact that: a) more than 70% of the collagen is native; b) it has a gelatin content of less than 4% by weight; c) at least 95% of the powder has a granulometry between 10 µm and 5 mm and it has an average granulometry between 250 µm and 1 mm.

Description

AREA OF INVENTION

[0001] The present invention belongs to the dietary field of satiety products. In particular, the invention relates to a dry collagen powder with satiating activity, which can be used as a food additive or incorporated into a food product to increase its satiating power. The invention also relates to the method for producing satiety powder and its application in dietary and / or medical treatments in humans or animals to increase satiety and reduce the feeling of hunger, thereby reducing food intake. The dry collagen powder of the invention is therefore useful in combating obesity.

[0002] The invention also relates to any use as a satiety agent or as a food supplement or as an ingredient in substitute products, foods for medical use or novel foods containing the collagen powder for weight loss, as well as with a food product containing the aforementioned collagen powder. BACKGROUND OF THE INVENTION

[0003] Obesity and its associated health problems are a global public health issue. Consequently, various strategies have been developed to combat it, including increasing metabolism, inhibiting digestion, and / or reducing calorie intake.

[0004] The use of satiety products or additives, designed to reduce appetite and thus minimize food intake, is part of a strategy to reduce calorie consumption. The feeling of fullness occurs naturally with food intake and lasts as long as the stomach is not emptied. Therefore, foods that remain in the stomach longer and are digested more slowly have a longer-lasting feeling of fullness. The swelling effect of food also promotes satiety by increasing pressure on the walls of the digestive tract.

[0005] Products with a saturation effect have been developed based on various compounds or substances and also based on different principles.

[0006] One approach to achieving a satiating effect is to increase the pressure on the stomach walls. For example, DE10161986 describes a composition for oral administration based on a solid, liquid, or gas that releases gas under the conditions of the stomach by increasing the pressure on the stomach walls and thus producing a satiating effect. However, this satiating effect is short-lived and therefore ineffective.

[0007] Another strategy is based on the use of polymers and / or polysaccharides with swelling capacity to provide the saturation effect.

[0008] For example, WO2009049105 describes a method to induce saturation by using polymers that increase the volume of what is chewed without increasing its energy density.

[0009] WO2004056375, in turn, refers to a substance for producing a saturation effect and reducing cholesterol levels, which contains little esterified polysaccharides and at least one polysaccharide that is able to swell and increase its volume.

[0010] Similarly, WO2004056393 describes a satiety agent based on polysaccharides containing polyuronic acid, which is able to swell in the stomach.

[0011] Other strategies aim to make the components of the satiety products more resistant to acid hydrolysis in the stomach, so that they exert their satiating effect in the duodenum. For example, EP 2651426 describes a product based on pea and / or wheat protein that is incorporated into an enteric-coated vehicle, so that the proteins reach the duodenum without hydrolyzing and exert their satiating function there.

[0012] Collagen or its derivatives have also been used to develop products or compositions with a satiating effect and as a means of combating obesity.

[0013] KR100417020 describes a composition with a saturating effect based on agar, alginate gel and gelatin from collagen hydrolysis.

[0014] EP0901792 describes a compressed and non-toxic vehicle that expands once in the stomach and has a sponge-like shape, thereby producing a satiating effect. This vehicle is derived from marine organisms such as sponges and has a partial collagen structure.

[0015] Document CN106509126 describes a weight loss and slimming beverage in the form of a water-reconstituted powder containing a complex mixture of several components and nutrients. One of these components is collagen powder.

[0016] EP2575496 describes a collagen powder with saturation properties. The collagen powder described in this document is characterized by being partially denatured, losing its native state, and containing a high percentage of gelatin.

[0017] The authors of the present invention have developed a collagen powder with satiating properties that can be used as a food or diet supplement, the main feature of which is that it has a greater swelling capacity than existing commercial products under pH conditions similar to those of the stomach during digestion, as well as greater resistance to enzymatic digestion, which makes it possible to prolong the satiating effect over longer periods.

[0018] The collagen powder of the present invention has not been subjected to any kind of denaturation process; therefore, it has zero or extremely low gelatin content. In fact, the collagen powder of the present invention has a structure very close to that of native collagen. SUBJECT OF THE INVENTION

[0019] The invention relates to a dry collagen powder with satiating properties, characterized in that: a) more than 70% of the collagen is native; b) it has a gelatin content of less than 4% by weight; c) at least 95% of the powder has a granulometry between 10 µm and 5 mm.

[0020] From now on, we will refer to this dry collagen powder interchangeably as the powder or the product of the invention.

[0021] Another object of the present invention is a food or dietary ingredient or food supplement comprising the powder of the invention. From now on, we will refer to this ingredient or food or dietary supplement interchangeably as the ingredient or food supplement of the invention.

[0022] Furthermore, a food product that contains, includes, or comprises the powder of the invention, or the ingredient or food supplement of the invention in its composition, is also an object of the invention.

[0023] Another aspect of the present invention is a method for producing the powder of the invention. We will refer to this as the method of the invention.

[0024] A final aspect of the invention is the use of the powder of the invention, the ingredient of the invention or the food product of the invention as a satiety product in humans or animals, as well as in the treatment and / or prevention of obesity. BRIEF DESCRIPTION OF THE FIGURES Fig. Figure 1: Results of the DSC analysis of a collagen powder sample according to the invention (1A), a Collapro® sample (1B), a Kapro® sample (1C), and a commercially available gelatin sample. The collagen powder sample according to the invention (1A) contains an endothermic peak that is characteristic of native collagen, while the other samples do not exhibit it, and therefore are not native collagen (1B, 1C, and 1D). Fig. 2: Beef after chemical treatment. Fig.3: Microscopic images at different dilutions showing details of the different fiber hierarchies that can be resolved by microscopic observation. Fig. 4: Images of the gels obtained in the Bloom test for the powder samples according to the invention (P1 and P2), for Kapro C®, for Kapro SF® and for a gelatin sample. Fig. 5: Image of the supernatant that appears during the gelation of one of the samples of the collagen powder according to the invention. Fig. 6: Average weight in grams of the animals during treatment in the different experimental groups. The data are presented as group mean ± SEM. Fig.7: Weight difference between the end and beginning of the average treatment in grams. Interaction test ANOVA 2x2 p < 0.005; t-Student * p < 0.0001 HFSPF CAS vs HFSPFCOL, * p < 0.0001 PF CAS vs PF COL. Data are presented as group mean ± SEM. Fig. 8: Average intake of the different experimental groups, expressed in g (A) and % relative to the control group (B). Effect of the interaction between the protocol and the type of supplemented protein on intake (C). ## p < 0.01 vs HFS; ### p < 0.001 vs HFS; p = 0.05 vs Casein 2h. Note: Group 1 (control) was not included in the statistical analysis, with Group 2 (HFS control) used as the reference group for one-way ANOVAs. Fig.9: Comparison of serum ghrelin levels for the collagen group at 2h versus the casein group at 2h. The group treated with collagen produced a significant decrease in ghrelin levels at 2 hours compared to the group treated with casein. DETAILED DESCRIPTION OF THE INVENTION

[0025] To facilitate understanding and clarify the meaning of certain terms in connection with the present invention, the following definitions are given: “Dry powder”: This is the powder according to the invention, which is obtained after a drying process to remove a large amount of water until a moisture content between 1% and 18% is achieved. Collagen: It is a protein whose basic unit molecule, called tropocollagen, consists of three chains of amino acids that are intertwined and linked together by hydrogen bonds. The tropocollagen units are grouped in various sequences, resulting in fibers that are in turn grouped into fiber bundles and finally into large fiber bundles. “Saturation properties”: refers to the ability of the powder according to the invention to satisfy the craving for food or drinks for a certain period of time. “Gelatin”: It is the product obtained after the denaturation of collagen, usually by applying high temperatures for a certain period of time. "Native collagen" refers to the property of a particular collagen to retain its native state, that is, to maintain the structure it naturally possesses in its source collagen, preferably the dermis. It can be said that the native state of collagen, or native collagen, is the opposite of denatured collagen. Bloom grade: This is a measure used to characterize the gelling strength of a product. It is the force required by a cylinder to penetrate a 6.67% collagen gel with water to a depth of 4 mm. It is a standard measure of the force required to cause deformation in a gel at a standardized concentration and temperature. It is commonly used to characterize gelatins marketed according to their gelling properties. There is a correlation between the Bloom grade and the gelatin concentration. The greater the amount of gelatin in a product, the greater its gelling strength and the higher its Bloom grade value. “Undigested collagen fraction”: This refers to the remaining collagen fraction that was not digested by pepsin under certain conditions, in particular, in connection with the present invention, those of the pepsin powder digestion test. "Pepsin powder digestion assay": refers to an analytical test that allows the determination of the degree of resistance of a specific collagen to digestion under acidic conditions using the enzyme pepsin, a protease (endopeptidase) that acts on the peptide bond by breaking it, thus releasing the amino and carboxyl groups. This test simulates digestion for 40 minutes at a controlled temperature using the enzyme pepsin and hydrochloric acid, mimicking the conditions of the stomach. "Percentage swelling": This is a measure used to assess the weight increase of collagen when subjected to the swelling test. The acid swelling test for powders involves placing the collagen powder in an acidic environment for two hours. After two hours, centrifugation is performed, separating the swollen collagen fibers, which are then weighed. The swelling is calculated as a percentage based on the weight difference. “Food or dietary supplement or ingredient”: In the context of the present invention, this refers to any food component containing the collagen powder of the invention and used in the manufacture or preparation of a food and still present in the finished product, even in a modified form. “Food product”: In the context of the present invention, this refers to all food products that are suitable for human consumption and can be sold to the public through chemical and / or physical industrial processes of treatment, handling, preservation and packaging, and which contain the collagen powder of the invention in its composition or the food supplement or ingredient of the invention. “Mechanical treatment”: In the context of the present invention, this refers to the set of processes designed to extract and reduce the size and length of collagen fibers from the collagen sources used. Mechanical treatments include crushing or grinding processes, which are carried out in a controlled manner and always with temperature control to avoid collagen denaturation. "Collagen source": refers to the raw material used for the extraction and subsequent treatment of collagen to produce the powder of the invention. In a preferred embodiment, the collagen source is of bovine, sheep, porcine, avian, or fish origin, or a mixture thereof. In a more preferred embodiment, the collagen source is hides of bovine origin from animals between 0 and 3 years of age, preferably less than 2.5 years of age. Collagen powder

[0026] The main aspect of the invention relates to a dry collagen powder with saturating properties, characterized in that it contains collagen with a high percentage of native collagen and very little or no gelatin.

[0027] In particular, the invention relates to a dry collagen powder with satiating properties, characterized in that: a) more than 70% of the collagen is native; b) it has a gelatin content of less than 4% by weight; c) at least 95% of the powder has a granulometry between 10 µm and 5 mm.

[0028] Preferably, the dry collagen powder consists of more than 75%, preferably more than 80%, and even more preferably more than 90% native collagen.

[0029] Preferably, the dry collagen powder has a gelatin content of less than 2.5 wt.%, preferably less than 1 wt.%, and even more preferably, it is free of gelatin.

[0030] Gelatin can be isolated in a dry powder from the fraction soluble in ammonium sulfate at a concentration of 10% and its subsequent centrifugation at 18,000 rpm. Using the biuret method, the protein content of the fraction can then be determined and expressed as a percentage. This fraction occurs when collagen is degraded. The precipitation of native collagen using salts that leave degraded collagen as the soluble fraction, including gelatin, was already described by Piez in 1967 (Soluble collagen and the components resulting from its denaturation, in: Treatise on Collagen (GNRamachandran, ed.) Vol. 1, p. 207, Academic Press, New York). We have found that when gelatin is added to a collagen sample, this method allows it to be accurately quantified.

[0031] The low percentage or absence of this fraction is directly related to the fact that the collagen is native, meaning that the collagen has not undergone any denaturation or hydrolysis treatment and is not degraded. The low percentage of gelatin in the powder of the invention is also derived from this.

[0032] One way to easily measure the degree of denaturation of the collagen powder of the present invention is DSC analysis. While the powder of the present invention shows a DSC profile of native collagen without detecting denatured collagen, and is characterized by a pronounced endothermic peak at 109.67 °C (see figure). Fig. 1A) and with a denaturation enthalpy of 46.65 J / g (sample moisture 15.5%), collagen powders from other previous commercial products such as Collapro® (see Fig. 1B) and Kapro® (see Fig.1C) a DSC profile without an endothermic peak, which is characteristic of native collagen. Their profiles show peaks around 82 °C, which correspond to one of the two peaks of gelatin, 82 °C and 53 °C (see Fig. 1D). They are therefore powders with a lot of denatured collagen (gelatin) and little or no detectable native collagen.

[0033] Therefore, the powder of the present invention consists of more than 70% native collagen (taking into account that a denaturation enthalpy of over 45 J / g °C is typical for native collagen, and taking into account that an enthalpy of 65 J / g °C for bovine collagen type I represents 100% native collagen), preferably more than 75%, more preferably more than 80%, and even more preferably more than 90%. Furthermore, the powder of the invention contains the ammonium sulfate-soluble fraction (% gelatin), the most degraded collagen, always less than 4% by weight, more preferably less than 2.5% by weight, even more preferably less than 1% by weight, and preferably in the absence of the gelatin fraction.

[0034] With regard to particle size, the collagen powder of the invention has a granulometry in which at least 95% of the particles lie between 10 µm and 5 mm, preferably at least 90% of the particles lie between 100 µm and 2 mm. The mean granulometry of the powder of the invention lies between 250 µm and 1 mm.

[0035] On the other hand, the collagen powder of the invention has exhibited physicochemical behavior that is particularly suitable for use as an additive with saturation properties in humans or animals.

[0036] On the one hand, it exhibits exceptional swelling capacity under pH conditions similar to those of the stomach. This swelling capacity is at least 32% higher than existing similar commercial products, including those described in EP 2575496. In particular, the powder of the invention has a swelling percentage in the acid swelling test of collagen powders of more than 1500%, preferably more than 1550%. This exceptional swelling capacity promotes a reduction in appetite or a feeling of satiety once the powder of the invention reaches the stomach, since the increase in its volume in the acidic pH environment of the stomach causes the chewed food to exert greater pressure on the stomach walls, thus increasing the feeling of fullness.

[0037] Similarly, the collagen in the powder of the invention, by maintaining a conformation very similar to that of its native state with virtually no denaturation or gelatinization, exhibits a much higher resistance to enzymatic digestion. This allows the satiating effect to last much longer, as the digestive system requires more time for its complete breakdown. The resistance to digestion of a collagen powder can be determined by the so-called pepsin digestion test. In this test, the undigested collagen fraction is measured after simulating digestion for 40 minutes at a controlled temperature using the enzyme pepsin and hydrochloric acid, which mimics the conditions of the stomach. In this test, the powder of the present invention exhibits an undigested collagen fraction of more than 30% after the pepsin digestion test.At best, commercial products comparable to the powder of the invention show no pepsin-induced digestive resistance results exceeding 15.6%. This means that the powder of the present invention exhibits at least twice the digestive resistance of other commercial products with which it has been compared.

[0038] Likewise, there is a method for characterizing the degree of gelation of a given collagen. This method is specified in the standard ISO 9665:1998(E), and the Bloom degree, or force, which expresses the force required by a cylinder to penetrate a 6.67% collagen gel with water to a depth of 4 mm, is measured. What is remarkable about the collagen powder according to the invention in relation to this test is not the Bloom value it achieves, but rather that the gels produced with the collagen powder according to the invention for this test exhibit a water supernatant that is not incorporated into the gel. This behavior is probably related to the low percentage of gelatin present in the collagen powder of the invention, as well as the high resistance of collagen to gelation under the test conditions (see Example 5).Gelatin has a high capacity to absorb water, but the absence of gelatin or its low percentage in the powder of the invention does not allow all the water to be absorbed, and therefore the supernatant likely appears in the Bloom test. It should be noted that the powder of the invention exhibits a higher swelling capacity than other existing powders when the test is performed in an acidic medium at pH values ​​similar to those of the stomach.

[0039] The powder of the invention is in dry form, and this implies that it has a moisture content between 1 and 16%.

[0040] The dry collagen powder of the invention has a particle size between 10 µm and 5 mm, preferably 80% of the particles are between 250 µm and 2000 µm. This facilitates, for example, that the product has a suitable size for its absorption as well as for its dispersion in a matrix with which it is to be combined for absorption.

[0041] Regarding the origin of the collagen, it can be derived from any source of animal collagen. It is also preferred that the collagen source be as fresh as possible, such as using the hides of recently slaughtered animals that have not been salted or stored for a long time. In a particular embodiment, the collagen is of bovine, sheep, porcine, avian, or fish origin, or a mixture thereof. In a preferred manner, the collagen is of bovine origin and from an animal between 0 and 3 years old, preferably less than 2.5 years old. Method for producing dry collagen powder

[0042] One subject matter of the present application is also a method for producing the collagen powder of the invention: a) Mechanical treatment of a collagen source that has been previously conditioned and acidified to reduce the size of the fibers until a collagen mass is obtained; b) Successive dilutions of the collagen mass from step a) at a temperature below 25 °C to obtain a concentrated collagen mass; c) Homogenization of the mass at a temperature below 25 °C and extrusion of the collagen mass from step b); d) Neutralization of the collagen mass from step c) with an alkaline agent to a pH between 6 and 9; e) Drying the collagen mass from step d) to a moisture content between 1% and 18%; and f) Grind the dry mass from step d) until a collagen powder is obtained.

[0043] The first step of the process aims to reduce the size of the collagen fibers until a viscous and homogeneous collagen mass is obtained, which is then processed in successive steps. For this purpose, the previously conditioned and acidified collagen source is mechanically treated. Any agent capable of mechanically reducing the fiber size can be used in step a) of the process, although the mechanical treatment is preferably carried out by grinding until a homogeneous mass is obtained.

[0044] As a step prior to this first step of the process according to the invention, the collagen must be conditioned and acidified. The conditioning and acidification treatment of the collagen source from step a) exhibits: i. chemical removal of hair from the collagen source, if the source used has it, ii. optionally one or more washes with oxidizing agent, iii. optionally an alkaline treatment to remove fat and other impurities from the collagen source; and iv. Acidifying the collagen source to a pH between 0.1 and 4.

[0045] Thus, conditioning the collagen source begins with the treatment of the collagen source itself, usually bovine, sheep, or pig hides containing hair that must be removed. This is generally achieved through chemical treatment with sulfur salts such as Na₂S and alkaline treatment with Ca(OH)₂.

[0046] Then, for proper conditioning, one or more washes with mild oxidizing agents such as diluted hydrogen peroxide can optionally be carried out to bleach and decontaminate animal hides by oxidizing the salt residues from the hair removal treatment.

[0047] Then, optionally, a treatment with an alkaline solution such as calcium hydroxide, sodium hydroxide or alkaline buffer systems with or without enzymes is carried out to soften particularly dense skin and to remove traces of fat and other residues or impurities present in the treated skins.

[0048] Finally, after the skins have been conditioned, the collagen source is subjected to acidification in order to be processed in the inventive method.

[0049] For acidification, the hides are subjected to successive washes in acidic media, for example with 33% HCl, to bring the pH value between 0.1 and 4. This pH value is suitable for the swelling of the hides, for the transformation of the material, and for making the fibers accessible in subsequent mechanical treatments.

[0050] Following mechanical treatment, step b) of the inventive process aims to obtain a mass, preferably with a collagen content between 3 and 7%, and to reduce the presence of gelatin as much as possible or even eliminate it completely. This step involves successive dilutions by milling and subsequent homogenization under controlled conditions, maintaining a temperature below 25°C. The result of this step is a mass with a high percentage of undenatured native collagen.

[0051] The mass is then extruded through an extrusion head to produce a collagen film with variable dimensions depending on the collagen mass being extruded. In one particular embodiment, the film can, for example, have dimensions of 12 cm in width x 0.6 mm in thickness. This produced film is then processed in the following steps.

[0052] Since the extruded film still has a strongly acidic pH, step d) involves neutralizing the film with an alkaline solution to a pH between 6 and 9. Any suitable alkalizing agent can be used, either a strong or a weak base. Neutralization is preferably carried out by immersion in a basic solution of sodium or potassium hydroxide. The ion exchange between the collagen film and the solution takes time, depending on the diffusion rate. Normally, times between 1 and 60 minutes are sufficient for neutralizing the collagen mass. Optionally, the neutralized film can be washed once or several times in preparation for the drying step.

[0053] Step e) involves drying the neutralized film to a moisture content between 1 and 16%. It is important that drying is carried out at a low temperature to avoid potential collagen denaturation. Drying can be performed using various methods, although ideally it is carried out in dryers using compressed air and at temperatures below 60°C until the product reaches the target moisture content, which can take anywhere from a few hours to 24 hours, depending on the dryer's capacity.

[0054] Finally, in step f), after drying, the film is crushed or ground until the collagen powder according to the invention is obtained. The grinding can be carried out by any method or means, although it is always ensured that the temperature during the process does not exceed the denaturation temperature. The grinding is preferably carried out in a mincer with temperature control so that it does not exceed 25 °C, and even more preferably by cryomilling using dry ice or a similar cryogenic substance. Cryomilling has the advantage that it makes it possible to reach the glass transition temperature of collagen, which makes it a fragile material that is prone to being fragmented into small particles during the grinding process, but without denaturing it.The dry collagen powder of the invention has a granulometry (measured with a laser diffraction particle size analyzer Malvern Instruments Mastersizer 3000) between 10 µm and 5 mm, and preferably with 80% of the particles between 250 and 2000 µm. Food or dietary supplement or ingredient

[0055] Another aspect of the invention is a food supplement or ingredient comprising the dry collagen powder of the present invention.

[0056] In addition to containing collagen powder, the dietary supplement or ingredient may contain other ingredients such as preservatives, antioxidants, colorings, flavorings, or any other component commonly used in the food industry.

[0057] The dietary supplement or ingredient of the present invention is intended to be incorporated into any foodstuff to increase its satiating power. In other words, the dietary supplement of the invention can be useful in connection with a dietary treatment to reduce appetite and lose weight for purely aesthetic purposes, but it can also be useful in medical treatments to combat obesity and insulin resistance in prediabetic or diabetic patients.

[0058] On the other hand, the food supplement of the invention is applicable in both human and animal food. food product

[0059] In connection with the previous aspect, a food product comprising the dry collagen powder of the invention, or the food supplement, or the ingredient of the invention, is also an object of the present invention.

[0060] The food product can be either a processed or an unprocessed product into which the powder, food supplement, or ingredient of the invention has been incorporated. In a specific way, and without claiming to be an exhaustive list, the food product can be a meat product, a dairy product, pasta, an egg product, a fish derivative, etc. Use of the dry collagen powder, the dietary supplement or ingredient and the food product of the invention

[0061] A final aspect of the invention is the use of the dry collagen powder of the invention, as well as the ingredient and the food product of the invention.

[0062] On the one hand, it is necessary to highlight the medical use of the powder of the invention and the products derived therefrom, which are applicable in both human and veterinary medicine, and on the other hand, its dietary use as a satiety product.

[0063] Thus, the powder of the invention can be used as a medicine, in particular it can be used as a medicine in the treatment and / or prevention of obesity and related diseases such as metabolic syndrome, diabetes and insulin resistance.

[0064] On the other hand, the powder, dietary supplement, ingredient, or food product of the invention has a use as a satiety agent in humans or animals to reduce appetite and with the ultimate goal of weight loss, provided there are no pathological processes that could themselves cause weight loss. This use could be considered a dietary or aesthetic use.

[0065] The powder or additive of the invention can be used by supplementing any meal of the day, be it breakfast, lunch or dinner, although preferably breakfast.

[0066] It can also be incorporated during the manufacturing process of processed foods to increase their satiating power, resulting in a food product according to the present invention. EXAMPLES

[0067] The following section illustrates how the powder according to the invention can be produced by means of several examples, as well as examples in which the powder is characterized by different parameters and these parameters are compared with other commercially available collagen powders. In vivo examples in rats are also provided, demonstrating the weight-loss and satiating effects caused by the collagen powder of the invention. These examples are presented as demonstrations but are not intended to limit the invention in any way. Example 1: Production of collagen powder

[0068] The reference raw material used to produce the powder was bovine hide. This raw material, as well as the collagen extraction methods, is well-established, which is why it is considered an ideal material for producing native collagen powder with saturation potential.

[0069] From the hides of young calves (approximately 2 years old), the inner layer of skin, known as the chorion, was separated from other layers of no interest, such as the grain, flesh, and fat layers. This chorion layer is rich in collagen protein; more precisely, more than 80% of the total protein is collagen protein and is called the split.

[0070] After obtaining the collagen-rich portion, the split, the chemical treatment of purification and collagen extraction was started, consisting of the following steps: 1. A pretreatment with hydrogen peroxide was carried out, which oxidized the remaining salts so that they could be dissolved and subsequently eliminated by washing. For this purpose, the raw material was added along with 35% hydrogen peroxide to a final concentration of 0.58% (% v / w) and mixed for 1 hour at room temperature. After the hydrogen peroxide treatment was complete, the liquid was drained and two 30-minute washes with water were performed. 2. Subsequently, controlled acidification with hydrochloric acid was carried out until a final concentration of 13.85% (% V / W) was reached, and the mixture was allowed to operate at room temperature for at least overnight. After this step, washing with water was performed to eliminate the excess acid from the previous step, yielding the rinds (see Fig. 2).

[0071] The native collagen fibers were extracted from these cattle using a two-stage mechanical grinding process: coarse grinding and fine grinding using a meat grinder (Gesame GP32). This yielded a heterogeneous paste with a high solids content. To convert this collagen paste into films of sufficient quality, a kneading and dilution step was necessary. Therefore, the collagen paste was mixed with the required amount of water and / or ice in a Z-mixer until a final solids content of between 4 and 7% was achieved. From this diluted mass, the collagen films were produced at room temperature using a laminator (Vansda, Thermal Laminator). The films were then immersed in a dilute ammonium hydroxide bath (0.03%) and washed with water. Finally, these wet films were allowed to dry at room temperature.

[0072] Finally, the dried films were crushed to obtain a powder with an average particle size of less than one millimeter.

[0073] For this purpose, the collagen films underwent cryomilling to obtain a fine particle with a flake-like appearance. A kitchen robot (Robot Coupe) equipped with a serrated blade was used for this cryomilling process. The film samples were introduced along with dry ice in a 1:3 ratio, and the material was milled at maximum speed for 5 minutes.

[0074] Thanks to the use of dry ice, it was possible to reach the glass transition temperature of the previously described collagen, making it a brittle material that could be fragmented into small particles during the grinding process without denaturing it. This resulted in a collagen powder with an average particle size between 250 and 1000 µm.

[0075] The analysis of the samples obtained was performed on both the intermediate and the final product. The following parameters were analyzed, and the results are shown in Table 1: Table 1: Values ​​of moisture, pH value, acid sources of the powder of the invention of Example 1 Collagen powder % humidity 13,7% PH value 7,6% % acid sources 1620% Example 2: Viewing collagen fibers using optical microscopy

[0076] Known quantities of the collagen powder samples obtained according to Example 1 were dispersed in 20 mM HCl to obtain a collagen concentration of 0.8%. This dispersion was prepared using ULTRA-TURRAX®. The powder was added gradually while stirring for approximately 5 minutes. Once the mixture was prepared, it was left to stand at 20°C for 24 hours.

[0077] The following day, the resulting mixture is processed to obtain various dilutions (between 1:10 and 1:100), and small fractions of the prepared dilutions are colored with the dye Sirius Red. Fig. Figure 3 shows microscopic images at various dilutions in great detail, revealing collagen fibers of different sizes and degrees of aggregation. Fibrils ranging from very small diameters (3B) to fibers larger than 20 µm (3C) and longer than 1 mm (3A) can be observed. Example 3: Ammonium sulfate-soluble fraction

[0078] A rapid method for quantifying the degree of degradation and hydrolysis of native collagen is to determine the fraction soluble in 10% ammonium sulfate after centrifugation of collagen powder that has been previously resuspended in 25 mM HCl at 18,000 rpm. This fraction, which contains gelatin, is particularly prevalent in degraded collagens.

[0079] The collagen protein content in the fraction is then quantified using the biuret method and the result is expressed as a percentage.

[0080] Table 2 shows the soluble fraction (in percent) of the collagen powder samples (P1 and P2) according to the invention in comparison to other commercially available collagen powders: Table 2: Percentage of the soluble fraction of two batches of the powder of the invention (P1 and P2) and of other commercially available collagen powders P1 P2 Kapro C® Kapro SF® Collapro® fine powder %G 0,0 2,0 14,5 20,2 18,1 Example 4: DSC analysis to determine the native collagen state

[0081] To analyze the native state of collagen, a DSC analysis was used on a sample of the powder according to the invention and on two other samples of commercially available collagen powders (Collapro® and Kapro®).

[0082] Samples ranging in size from 3 to 25 mg, depending on the water content, were weighed and encapsulated in hermetically sealed aluminum crucibles to prevent water evaporation. The sealed crucibles containing the samples were then weighed.

[0083] The scan was performed using DSC equipment at a rate of 10 °C / min. The crucibles were reweighed after the scan to verify that they had not lost any sample or water. The scan was started at a temperature at least 25 °C lower than the temperature expected for the first transition (e.g., denaturation), in this case at 15 °C. For thermogram analysis and interpretation using the software, the key factors for collagen are: denaturation temperature (triple helix stability), denaturation enthalpy (amount of crystalline, ordered, native collagen), temperature and magnitude of the Tg (glass transition temperature), and the shape of the thermogram. Collagen denaturation is an endothermic process, meaning a more or less broad peak appears above the baseline.The temperature at which this peak is observed depends on the water content of the collagen sample, which is 110-120 °C for collagen with a low percentage of water.

[0084] In the Fig. Figures 1A-C, which show the result of the DSC analysis, show how the powder according to the invention exhibits a characteristic peak of native collagen ( Fig. 1A), while the samples from Collapro® ( Fig. 1B) and Kapro® ( Fig. 1C) show a clear profile of denatured collagen without the characteristic endothermic peak of native collagen. Their profiles show peaks around 82 °C, as in the case of gelatin, which also has a peak at 82 °C, in addition to another at 53 °C (see Fig. 1D). Therefore, samples 1B and 1C are powders with a high amount of denatured collagen (gelatin) and a low amount of native collagen. Example 5: Bloom test (ISO9665 standard)

[0085] To observe the differences between the various powdered collagen samples, gel strength measurements are performed using the Bloom grade test.

[0086] This test makes it possible to analyze how different collagen samples gel.

[0087] Therefore, 7.5 grams of each sample to be tested were dissolved in 105 ml of distilled water, corresponding to a concentration of 6.67%, by stirring with a glass rod. Next, to improve the uniformity of the samples and before heating to 65 °C, all dispersions of the different collagen powders were homogenized in an Ultra-Turrax at 15,000 rpm for 1 minute.

[0088] After homogenizing the dispersions in the Ultra-Turrax, they are heated in a water bath at 65 ± 1 °C for 15 min, with each dispersion being gently stirred a few times with a glass rod during this time.

[0089] Next, the containers are covered with Parafilm (glass with an inner diameter of 59 ± 1 mm and a height of 85 mm, specifically designed for measuring Bloom degrees) and left to stand at room temperature for 1 hour. They are then placed in a water bath at 10.0 ± 1 °C for 17 ± 1 hour. After 17 hours, the containers with the gelatin that has already formed are removed from the water bath and warmed to room temperature for 1 hour. Finally, measurements of each sample are performed in a texture analyzer at 20 °C.

[0090] The measurement is carried out by penetrating a cylinder (0.5 " radius cylinder probe P / 0.5R) into the gelatin contained in the glass using the 5 kg cell and with a deformation rate of 0.5 mm / s according to the ISO standard.

[0091] Once a force of 4 g is reached, the cylinder penetrates the gel to a depth of 4 mm, and the force required to reach this depth is the so-called “bloom strength” (g) of the gel.

[0092] The results for the different samples tested are shown in Table 3: Table 3: Bloom grade of two batches of the powder of the invention (P1 and P2), Kapro C® and Kapro SF® and a commercially available gelatin P1 P2 Kapro C® Kapro SF® gelatin Bloom grade 43 45 112 67 108

[0093] Fig. Figure 4 shows the gels obtained for each of the tested samples. With regard to the samples of powders P1 and P2 according to the invention, it should be noted that once gelled, the samples showed a liquid supernatant of approximately 1 cm thickness (see Figure 4). Fig.5) To perform the Bloom test measurement, it was necessary to remove the supernatant, since the test only begins when the flask reaches a minimum resistance of 4 g, and the water does not offer this resistance. The low gelling ability of the powder according to the invention is therefore noteworthy. Although the sample is heated to 65 °C for 15 minutes during the analytical process, it appears that this does not produce a complete gelatinization effect of the collagen in the powder according to the invention, probably due to the high resistance to denaturation of the collagen according to the invention, which in turn confirms its similarity to collagen in its native state. Example 6: Pepsin Digestive Test

[0094] The pepsin digestion test allows us to simulate the digestive conditions in the stomach and to determine the degree or percentage of undigested collagen at the end of the test.

[0095] Selective enzymatic hydrolysis of collagen leads to intermediate species such as peptones, peptides and even amino acids.

[0096] The pepsin enzyme is a protease (endopeptidase) that acts on the peptide bond, breaking it and thus releasing the amino group and the carboxyl group.

[0097] Two samples according to the invention (P1 and P2) as well as samples of Kapro C®, Kapro SF® and Collapro® were tested. 0.6 grams per sample were weighed.

[0098] The gastric sensitizer pepsin solution was prepared at a concentration of 3.2 g / L and left to incubate for 15 minutes to activate the pepsin. 40 ml were added for each 0.6 g sample and the solution was allowed to incubate at 37 °C for 40 minutes.

[0099] The reaction was quenched with 1N NaOH. It was filtered through 5-9 micron filters to retain the undigested solids on the filter and dried in a vacuum oven at 160 °C for 1 hour.

[0100] The results (see Table 4) were expressed as the percentage of undigested collagen compared to initial collagen in dry extract. The following calculations were performed for this purpose:

[0101] This technique requires taking into account the percentage of moisture in the collagen powders. - Calculation of the initial collagen dry weight: Initial collagen dry weight (Psi) = Pcol−(Pcol×% moisture / 100) - Calculation of % undigested solids: Psf=Ps−Pp−Pf % undigested solids = (Psf × 100) / Psi where: Psi = initial collagen dry weight Pcol = Collagen sample weight (g) Psf = final solid dry weight (g) Ps = dry weight (g) Pp = plate weight (g) Pf = Filter weight (g) Table 4: Percentage of undigested collagen from two batches of the powder of the invention (P1 and P2), Kapro C® and Kapro SF® and Collapro® P1 P2 Kapro C® Kapro SF® Collapro® fine powder %undigested 37 42 10 2 0

[0102] A greater resistance to enzymatic digestion of the powder of the invention is clearly observed compared to other commercial products, which in some cases are completely digested. It is assumed that the increased resistance to enzymatic degradation in this "gastric simulator" causes a delay in digestion and thus increases the satiating effect of the product. Example 7: Acid swelling test

[0103] Swelling (SW) measures the water retention capacity of collagen in an acidic medium under pH conditions similar to those of the stomach. Two samples of the powder according to the invention (P1 and P2) and three samples of commercially available collagen powders (Kapro C®, Kapro SF® and Collapro®) were tested.

[0104] For this purpose, 0.6 g of collagen powder from each sample was weighed into a Falcon tube suitable for centrifugation, and 30 ml of 0.1 N HCl was added. After stirring, the mixture was allowed to stand at room temperature for 2 h. It was then centrifuged at 18,000 rpm at 23 °C for 10 minutes. The supernatant was removed, and the pellet was collected for immediate weighing. The sample was then dried in an oven at 160 °C until the weight constant was reached.

[0105] The % swelling was calculated using the following formula: %SW=((PH / PS)×100)−100 where: - pH: Wet weight - PS: Dry weight

[0106] The results of the source test are shown in Table 5: Table 5: SW source results (%) of two batches of the powder of the invention (P1 and P2) and two samples of commercially available collagen powders (Kapro SF® and Collapro®) P1 P2 Kapro SF® Collapro® fine powder % SW 1978 1842 1468 1051

[0107] These results show that the collagen powder samples according to the invention exhibit a significantly higher water retention capacity than commercially available products. This will directly affect the saturation power of the powder according to the invention, which is greater than that of other commercially available powders. Example 8: In vivo test for weight loss

[0108] In this test, a large quantity of collagen powder corresponding to P1 and P2 of the invention was evaluated, selected for its potential effects on weight loss in an in vivo model using rats that had previously developed obesity through the intake of a high-fat, high-sugar (HFS) diet.

[0109] The proposed work scheme for this test was divided into 3 steps. The first step was a 4-week acclimatization of the animal model used, Wistar rats weighing approximately 50-75 grams, which were given a control diet (Envigo, diet 2014S).

[0110] The second step began when the animals reached an average weight of 200 grams and lasted 8 weeks. It consisted of administering a powdered diet high in fat and sugar (Research Diets, Diet 12451) with the aim of inducing a dietary obesity model, except for the control group, which remained on a powdered control diet.

[0111] Finally, the third step was the treatment / intervention, which consisted of administering the following ad libitum diets to the experimental groups (throughout the day): a control diet; a high-fat, high-sugar, protein-free diet (HFSPF); and a protein-free diet (PF). The latter two were supplemented with protein at 14.3% w / w of the total diet weight. In the casein groups, the diet was supplemented with 14.3% casein, and in the collagen groups, with 10% collagen plus 4.3% casein until 14.3% protein was added. The dietary supplementation percentages were selected based on the reviewed literature and previous pilot tests. The collagen experimental groups were also supplemented with tryptophan until the amount of this amino acid in the casein group was balanced, given the absence of this essential amino acid in collagen (Table 6). Table 6: Percentage of essential amino acids (EAs) in the control and treatment diets. Tryptophan was supplemented based on the amount contained in the casein-only group. EA data were extracted from the composition provided by the manufacturer of the control diet. For casein and collagen EA data, mean values ​​were estimated from information on various marketed products. Essential amino acids (AE) AE control AEKasein % AEWithCasein AEKollagen %AEWithCollagen %Total AE Casein (4.3%) +Collagen10% Diet Supplemented amount in collagen diet Threonine 0,50% 4,40% 0,15% 1,72% 1,20% 1,35% No Leucin 1,40% 8,30% 0,42% 2,62% 1,83% 2,25% No Isoleucine 0,60% 5,50% 0,18% 1,36% 0,95% 1,13% No Valin 0,70% 6,50% 0,21% 2,17% 1,52% 1,73% No Phenylalanine 0,70% 4,50% 0,21% 1,90% 1,33% 1,54% No Methionine 0,30% 2,50% 0,09% 0,54% 0,38% 0,47% No Lysine 0,70% 7,40% 0,21% 3,07% 2,15% 2,36% No Histidine 0,40% 2,80% 0,12% 0,72% 0,50% 0,62% No Trypophan 0,20% 1,30% 0,06% 0,00% 0,00% 0,06% 0,14%

[0112] After treatment, the animals were slaughtered and various tissues were extracted for later analysis.

[0113] The study was conducted for two consecutive weeks, during which the intake and weight of the animals were checked every 2-3 days. A total of 55 animals were used, housed in individual cages and randomly assigned to the five experimental groups described below (n = 11 rats / group): - Group 1: Control group (CT). Administration of the 2014S diet (14.3% w / w protein), ad libitum throughout the day. - Group 2: High-fat and high-sugar protein-free + casein diet group (control protein; HFSPF CAS). Administration of Research Diets diet D12451px07, supplemented with casein in an amount corresponding to 14.3% of the dry weight of the total daily diet. - Group 3: High-fat and high-sugar protein-free + collagen diet group. HFSPF COL. Administration of Research Diets diet D12451px07, supplemented with collagen in an amount equal to 10% of the dry weight of the total daily diet, plus 4.3% casein to match protein intake to that of the control. - Group 4: Protein-free + casein diet group (control protein, PF CAS). Administration of the Envigo 93328 diet supplemented with casein in an amount corresponding to 14.3% of the dry weight of the total daily diet. - Group 5: Protein-free + collagen diet group (PF COL). Administration of the Envigo 93328 diet, supplemented with collagen in an amount corresponding to 10% of the dry weight of the total daily diet, plus 4.3% casein to match protein intake to that of the control. RESULTS: WEIGHT ASSESSMENT

[0114] From the third day of treatment onwards, a particularly drastic decrease in the weight of the rats was observed in the groups that were supplemented with collagen in both the HFSPF diet and the PF diet ( Fig. 6 and Fig. 7) In fact, the weight loss in the groups supplemented with collagen resulted in them having a similar weight to the control group at the end of the study ( Fig. 6) On the other hand, no major weight changes were observed in the groups supplemented with casein ( Fig. 6 and Fig. 7).

[0115] To rule out dehydration as the cause of the weight loss, since undigested collagen could retain water, the animals' water consumption was measured. However, no significant differences were observed between the various experimental groups. Therefore, dehydration was ruled out as the cause of the weight loss. Furthermore, the animals showed no external signs of dehydration, nor any other obvious changes. ASSESSMENT OF THE WEIGHT OF ORGANS AND TISSUES

[0116] During the slaughter of the animals, various organs were examined macroscopically, extracted, and weighed for later analysis. Additionally, samples were collected from several specimens for toxicological analysis.

[0117] Thus, various types of adipose tissue were collected, designated as epididymal, mesenteric, brown, retroperitoneal, and subcutaneous fat relative to the animal's weight. As shown in Table 7, all fat deposits (except brown) showed significant differences with treatment, with a lower percentage of these tissues relative to the animal's weight in the collagen groups than in the casein groups. Therefore, it can be concluded that the collagen treatment significantly reduced the percentage of fat mass, affecting virtually all fat deposits in the animals. Table 7: Weight in grams of organs and tissues relative to the animal's weight in the experimental groups supplemented with collagen and casein on the HFSPF and PF diets. ANOVA 2x2 test. Data are presented as group mean ± SEM. HFSPFCAS HFSPFCOL PF CAS PF COL diet Treatment interaction Epididymal fat 13,30 ±0,91 10,42 ±1,24 12,03 ±0,84 10,57 ±0,66 0,0254 Mesenteric fat 5,815 ±0,551 4,562 ±0,321 4,849 ±0,321 4,364 ±0,392 0,0389 Brown fat 0,760 ±0,038 0,792 ±0,067 0,745 ±0,61 0,884 ±0,078 Retro G. 15,52 ±1,15 11,48 ±1,06 14,26 ±0,67 11,07 ±0,87 0,0005 Subcutaneous fat 7,967 ±0,737 5,821 ±0,517 7,678 ±0,403 5,546 ±0,481 0,0004 Visceral fat 34,64 ±2,45 26,46 ±2,54 31,14 ±1,58 26,01 ±1,83 0,0034 Total fat 42,60 ±3,07 32,28 ±2,99 38,81 ±1,96 31,55 ±2,23 0,0017 Soleus 0,151 ±0,011 0,158 ±0,010 0,1573 ±0,011 0,171 ±0,007 Gastrointestinal (English: Gastro) 2,686 ±0,112 2,373 ±0,085 2,324 ±0,025 2,447 ±0,104 0,0204 liver 9,550 ±0,185 9,230 ±0,312 9,674 ±0,383 10,89 ±0,569 spleen 0,661 ±0,036 0,534 ±0,019 0,656 ±0,038 0,704 ±0,021 0,0060 kidney 2,270 ±0,078 2,160 ±0,045 2,237 ±0,062 2,356 ±0,035 0,0235

[0118] When examining visceral fat (the sum of all deposits located in the abdominal cavity) and total fat (the former plus subcutaneous fat), both as a percentage of body weight and in total grams, a decrease was also observed in those supplemented with collagen. Therefore, it is concluded that the weight loss in the collagen-supplemented groups was primarily due to the loss of fat. CONCLUSION:

[0119] Supplementation with 10% collagen in the diet induced a decrease in body weight, which is attributable to a decrease in fat mass, without affecting muscle mass or observing symptoms of dehydration or gastrointestinal changes, nor macroscopic changes of toxic effects. Example 9: In vivo test for the saturation effect

[0120] The capacity of the collagen powder of the invention to act as a saturating agent was evaluated, as well as its potential metabolic effects in an in vivo model with rats that had previously developed obesity through the intake of a high-fat, high-sugar diet (HFS).

[0121] Wistar rats with an approximate initial weight of 50-75 grams were used. The proposed procedure for this experiment was divided into 3 steps:

[0122] The first step, acclimatization, lasted 3 weeks after the animals included in the study were admitted, during which time a control diet was administered (Envigo, diet 2014S).

[0123] The second step, induction of the obesity model, began when the animals reached an average weight of approximately 180 grams and lasted 7 weeks. It consisted of administering a high-fat, high-sugar diet (HFS-Research Diets, Diet 12451) with the aim of inducing obesity and insulin resistance in the animals, except for the control group (Group 1), which remained on the control diet. This group served as a control for the obesity model used.

[0124] Finally, when the animals fed the HFS diet showed significant differences in weight and body composition compared to the control group, the third step of the study began, consisting of the treatment / intervention phase. During this phase, all animals received a control diet, and two clearly differentiated experimental strategies were conducted in parallel to evaluate the potential satiating effect of collagen. The first strategy involved ad-libitum feeding of the experimental groups (throughout the day). The rats' normal diet was supplemented with a collagen / casein amount corresponding to 7.5% of the total dry weight of the diet. Along with the diet, the animals had free access to water to promote an increase in collagen volume and thus enhance satiety due to greater gastric filling. For the second experimental strategy, after a 12-hour fasting period (o / n), 4 g of diet containing the satiating agent or its control (e.g., powdered collagen or casein, representing 7.5% w / w of the total daily intake) was administered, and the rat was allowed to eat for 1.5 hours to ingest as much as possible. After this time, and once satiating agent intake was confirmed, the diet was administered ad libitum for the next 10.5 hours. Throughout this period, the animals had free access to water. Once intake was assessed, food and water were withdrawn for 12 hours (o / n) to keep the animals fasting again until the following day, when the procedure was repeated.

[0125] Following these two experimental strategies and including the corresponding control groups (groups 1 and 2), a total of 60 Wistar rats were used, arranged in individual cages, which were distributed into 6 groups (10 animals per group) according to the following scheme, developed over seven consecutive weeks, during which intake was monitored (daily). - Group 1: Control diet group. Administration of the Envigo 2014S diet ad libitum throughout the entire trial. - Group 2: Obese control group. HFS diet (diet D12451, high fat and sucrose) ad libitum for 7 weeks and control diet (2014S) for 7 weeks thereafter. - Group 3: Group treated with 24-hour casein (Casein 24h). 7 weeks HFS diet (D12451). Then, control diet (2014S), supplemented with casein corresponding to 7.5% w / w of total daily intake, for a further 7 weeks. - Group 4: Group treated with Collagen 24h (Collagen 24h). 7 weeks HFS diet (D12451). Then, control diet (2014S), supplemented with collagen corresponding to 7.5% w / w of total daily intake, for another 7 weeks. 3 / 11 Group 5: Group treated with casein only in the morning (casein 2h). 7 weeks on the HFS diet (D12451). Then, a control diet supplemented with casein (corresponding to 7.5% w / w of total intake, but only between 8 and 9:30 a.m.). The rest of the day (9:30 a.m. - 8:00 p.m.) was a control diet (2014S) ad libitum. Fasting overnight, and this protocol was repeated the following day. - Group 6: Group treated with collagen only in the morning (collagen 2h). 7 weeks HFS diet (D12451). Afterwards, control diet supplemented with collagen (corresponding to 7.5% w / w of total intake, but only between 8 and 9:30 a.m.). The rest of the day (9:30 a.m. - 8:00 p.m.) control diet (2014S) ad libitum. Fasting overnight, and this protocol is repeated the following day. RESULTS: FOOD INTAKE ASSESSMENT

[0126] Individualized intake data for each of the 60 rats included in the study were recorded daily throughout the trial. During the 7-week intervention, intake across the different groups ranged from approximately 17 g to 21 g per day. The intake trend between groups remained relatively constant throughout the 7 weeks, with the group receiving the diet supplemented with 7.5% w / w collagen early in the morning (group 6) exhibiting lower intake levels. Significant differences in intake were observed between the groups when the data were analyzed using an ANOVA that compared all groups. Specifically, the groups following a feeding pattern with protein supplementation in the first hour of the day (groups 5 and 6) consumed a lower amount of the diet than the rest of the groups. Fig. 8B).

[0127] Of particular interest is the ANOVA 2x2 analysis of the uptake data from groups 3-6 (casein 24h, collagen 24h, casein 2h, and collagen 2h). This analysis allows us to observe the effect of the two main factors: the type of treatment and the type of protein the animals received. The statistical analysis of the data in Fig.The data presented in Figure 8C indicate a significant interaction between protocol and protein factors. The interaction graph shows that only in the groups following the 2-hour protocol (treatment administration early in the morning) was intake higher when the protein was casein than when it was collagen. Analyses allowing us to infer the simple effects confirm this hypothesis, and significant differences in intake are observed between the casein 2h group and the collagen 2h group when the animals followed the dietary supplementation protocol early in the morning. This result supports the hypothesis of a potential saturation effect of collagen when administered early in the morning and is consistent with previous findings that this group also exhibited the least weight gain.

[0128] Once the differences in uptake as a function of the protein supplemented in the diet were observed when the rats received the feeding pattern with the supplement early in the morning, the next goal was to try to clarify the possible molecular mechanisms underlying the saturation effect of collagen that is observed.

[0129] For this purpose, blood samples were taken from the tails of animals belonging to groups 5 and 6 (collagen 2h and casein 2h) under fasting conditions after ingestion of the protein-supplemented diet and 3 hours after ingestion of the supplemented diet. The samples were then processed and plasma levels of ghrelin (an orexigenic hormone that promotes intake) were analyzed.

[0130] It is particularly interesting to see that serum ghrelin levels in the collagen 2h group decreased significantly in the hours after initiation of treatment compared to the casein 2h group (see Fig. 9). EVALUATION OF BIOCHEMICAL PARAMETERS

[0131] Serum and plasma samples were obtained from the slaughtered animals for subsequent analysis of the following biochemical parameters of interest (Table 9). These blood samples were collected from the rats after a 12-hour fast. As shown in Table 9, no relevant changes in the animals' overall metabolic levels were observed as a function of the type of protein supplemented to the diet when the animals followed the same feeding pattern. Statistically significant differences were observed only in HDL cholesterol and total cholesterol levels, which were reduced in the collagen 2h group compared to the casein 2h group. However, these differences were neutralized in the analysis of the total cholesterol / HDL ratio, so this result has no particular physiological relevance.

[0132] Furthermore, it should be noted that no abnormal values ​​of transaminases (ALT and AST) were observed in any of the study groups, which makes it possible to rule out a toxicological effect.

[0133] Analysis using ANOVA 2x2 of the various parameters analyzed, including the protocol and the type of protein the animals received as key factors, revealed significant differences only for cholesterol, HDL cholesterol, triglycerides, and insulin markers, depending on the protocol followed. The type of protein received was a determining factor with respect to ALT and AST levels. For the two protocols performed, the animals treated with collagen showed lower levels of both markers (ALT and AST), which is very promising, as it is known that high protein intake (e.g., casein) tends to lead to an increase in transaminase levels. Table 8: Biochemical parameters of the experimental groups included in the study. Data are presented as group mean ± SEM. * p < 0.05 vs HFS; *** p < 0.001 vs HFS; ## p < 0.01 vs Casein 2h; bp < 0.05 vs Col 24h; bb p < 0.01 vs Col 24h. Note: Group 1 (control) was not included in the statistical analysis, with Group 2 (HFS control) used as the reference group for one-way ANOVAs. control HFS CAS24h 24H COL CAS 2H COL 2H Glucose (mg / dl) 121,21 ±1,28 129,07 ±1,88 131,13± 2,31 132,18 ±2,40 129,68 ±2,28 126,56 ±3,17 Insulin (µg / L) 0,75 ± 0,07 1,34 ±0,18 1,47 ±0,19 1,50 ± 0,28 1,47 ± 0,25 1,29 ± 0,28 HOMA-IR 5,59 ± 0,51 10,72 ±1,57 11,76 ±1,46 12,21 ±2,34 11,87 ± 2,12 10,33 ± 2,60 Cholesterol (mg / dl) 71,20 ±2,84 76,60 ±3,66 76,50 ±2,24 86,90 ±2,89 71,70 ± 2,97 62.30 ±2.81*,bb HDL-COL (mg / dl) 23,51 ±0,88 23,34 ±0,72 24,58 ±0,56 25,72 ±0,66 25,16 ±0,88*** 21.49 ± 0.74##, bb Total cholesterol / HDL cholesterol 3,03 ± 0,06 3,27 ±0,07 3,11 ±0,06 3,38 ± 0,05 2,85 ±0,05*** 2,90 ±0,07*** AIP Index 0,45 ± 0,04 0,58 ±0,04 0,52 ±0,04 0,51 ± 0,03 0,39 ± 0,05* 0,40 ± 0,05* TG serum (mg / dl) 69,40 ±8,19 91,10 ±7,56 84,40 ±8,30 86,20 ±7,54 65,20 ± 8,23 57,00 ± 7,65* TG liver (mg / dl) 4,46 ± 0,67 5,77 ±0,78 6,09 ±0,51 6,08 ± 0,46 4,82 ± 0,46 4,88 ± 0,52 ALT (U / L) 37,22 ±1,81 43,35 ±2,27 45,64 ±2,25b 36,79 ±1,68 41,35 ± 1,98 37,04 ± 2,17 AST (U / L) 150,31 ±8,480 183,34 ±15,12 187,56± 12,43 148,43 ±10,98 178,21 ±11,65 164,64 ±16,43 CONCLUSION:

[0134] At the macroscopic and biochemical levels, no adverse effects were observed that could have been caused by collagen intake. In fact, transaminase levels were lower in animals supplemented with collagen than in those supplemented with casein. The results of the present study show that the collagen powder of the invention promotes an inhibition of the animals' total daily intake when supplemented in a single dose early in the morning. These saturation effects of collagen could be caused, at least in part, by its greater gastric swelling capacity, which promotes lower postprandial ghrelin release and thus inhibits food intake. QUOTES INCLUDED IN THE DESCRIPTION

[0000] This list of documents cited by the applicant was automatically generated and is included solely for the reader's convenience. The list is not part of the German patent or utility model application. The DPMA accepts no liability for any errors or omissions. Cited patent literature

[0000] DE 10161986

[0006] WO 2009049105

[0008] WO 2004056375

[0009] WO 2004056393

[0010] EP 2651426

[0011] KR 100417020

[0013] EP 0901792

[0014] CN 106509126

[0015] EP 2575496 [0016, 0036] Cited non-patent literature

[0000] Piez described in 1967 (Soluble collagen and the components resulting from its denaturation, in: Treatise on Collagen (GNRamachandran, ed) Vol. 1, P.207, Academic Press, New York

[0030]

Claims

[1] Dry collagen powder, characterized by , that: a) more than 70% of the collagen is native; b) it has a gelatin content of less than 4% by weight; c) at least 95% of the powder has a granulometry between 10 µm and 5 mm and it has an average granulometry between 250 µm and 1 mm. [2] The powder according to claim 1, characterized by that it is composed of more than 80%, preferably more than 90%, and even more preferably more than 95% native collagen. [3] The powder according to any one of the preceding claims, characterized by that it has a gelatin content of less than 2.5 wt.%, preferably less than 1 wt.%, and even more preferably the absence of gelatin. [4] The powder according to any of the preceding claims, which has an undigested collagen fraction of more than 30% according to the pepsin powder digestion test. [5] The powder according to any of the preceding claims, wherein the swelling percentage in the acid swelling test of collagen powders is more than 1550%. [6] The powder according to any one of the preceding claims, wherein the collagen is of bovine, sheep, porcine, avian, fish origin or a mixture thereof. [7] The powder according to claim 6, wherein the collagen is of bovine origin and is from an animal between 0 and 3 years of age, preferably less than 2.5 years of age. [8] A food or food supplement comprising the collagen powder according to any one of claims 1 to 7. [9] A food product comprising a powder according to any one of claims 1 to 7 or a food supplement according to claim 8.

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