EP3737736A1 - Process for producing oils and defatted meal by means of solid/liquid extraction - Google Patents

Abstract

The invention relates to a process for producing an oil and defatted meal by means of solid/liquid extraction. The process comprises a step of solid/liquid extraction using a solvent containing 2-methyloxolane and water.

Description

Description

Title: Process for the production of defatted oils and oilcakes by solid / liquid extraction

Technical area

[1] The invention relates to the field of production of defatted oils and cakes from a biological substrate by a solid / liquid extraction process.

Context of the invention

[2] Oils play an important role in the food, pharmaceutical, nutraceutical and cosmetic industries. Recently attention has focused on the production of oils rich in polyphenols. Indeed, according to recent studies, oils rich in polyphenols have beneficial effects on health, in particular on the skin and against cardiovascular diseases and cancers. Recent industrial developments have therefore focused on the production of oils rich in polyphenols.

[3] There are two main known categories of oil production processes, the mechanical process which makes it possible to obtain a virgin oil and the process by solid / liquid extraction making it possible to produce an oil from a solid substrate. Conventionally, the solid / liquid extraction method comprises a solid / liquid extraction step involving a solvent to obtain a liquid fraction comprising an oil and the solvent.

[4] The two mechanical and solid / liquid extraction processes can be combined, namely a first mechanical process followed by a liquid solid extraction, the solid being constituted by the solid residue from the mechanical process.

[5] Solid residues, also called cake, are mainly used in animal feed, and increasingly in human food, mainly as a protein supplement, but also as a metabolizable energy source. [6] It is important that these meals are defatted because this promotes their conservation over time (limitation of the risk of rancidity) and facilitates the isolation of proteins for animal feed. In addition, the fat disrupts the digestion of animals, in particular cattle.

[7] Currently, hexane is the most widely used solvent in the solid / liquid extraction process because it has advantageous properties. Indeed, hexane allows a high yield extraction of the oil (> 97%) and allows to leave less than 3% of residual oil in the solid substrate, which prevents the latter from becoming rancid. Hexane is easy to separate from the extracted oil, it has a suitable boiling point (ie high enough to limit losses during extraction but low enough to limit energy consumption during its separation from oil extracted and during its recycling), it is stable and has good lipid selectivity.

[8] However, hexane has significant drawbacks. It is of petroleum origin, it is neurotoxic and is classified as toxic for reproduction of category 2. It is also toxic for the aquatic environment of category 2.

[9] In addition, analyzes of the oils obtained by solid / liquid extraction with hexane have shown that the process using hexane does not make it possible to produce an oil rich in polyphenol.

[10] Alternative solvents to hexane have been tested. For example, the article by Anne-Gaëlle Sicaire et al. "2-Methyltetrahydrofuran: Main Properties, Productions Processes, and Application in Extraction of Natural Products" on pages 253-268 of "Alternative Solvents for Natural Products Extraction" describes the solid / liquid extraction of blackcurrant buds with anhydrous 2-methyloxolane to get color and flavor.

[11] The present invention therefore aims to provide an industrial solid / liquid extraction process which makes it possible to produce a crude oil rich in polyphenol and / or a defatted oil cake. [12] Surprisingly, the Applicant has developed a process which meets this need.

Statement of the invention

[13] According to a first aspect, the invention relates to a process for producing a crude oil rich in polyphenol from a biological substrate comprising a step of:

a) solid / liquid extraction of the biological substrate with a solvent to obtain on the one hand a liquid fraction comprising the crude oil and the solvent and on the other hand a solid residue,

characterized in that,

the solvent comprises 2-methyloxolane and water, and

the mass percentage of water in the solvent during step a) of extraction is from 0.3% to 20%.

[14] For the purposes of the present invention, the term "solid / liquid extraction" means obtaining a liquid fraction and a solid residue from a solid biological substrate using a liquid solvent as solvent for extraction. Typically solid / liquid extraction can be carried out by decoction, infusion, digestion, percolation, leaching or maceration. A liquid / liquid extraction step is not a solid / liquid extraction step within the meaning of the present invention because it does not use a solid biological substrate; moreover, the physico-chemical characteristics involved in a solid / liquid extraction step and in a liquid / liquid extraction step are different.

[15] For the purposes of the present invention, "crude oil" means an unrefined oil, ie which has not undergone a chemical or mechanical refining step, also called physical, after its extraction by a solid / liquid process .

[16] Advantageously, the method according to the invention makes it possible to produce a crude oil richer in polyphenol than the crude oil produced by the methods using anhydrous hexane or 2-methyloxolane. [17] For the purposes of the present invention, "crude oil rich in polyphenol" means a crude oil comprising one or more polyphenols and whose mass concentration of polyphenols is greater than or equal to 100 ppm, in particular from 320 ppm to 2000 ppm, more particularly from 350 ppm to 1500 ppm, more particularly still from 400 ppm to 1200 ppm.

[18] For the purposes of the present invention, the term "polyphenol" means a class of molecules characterized by the presence of several phenolic groups arranged in complex structures.

[19] Crude oil may also include a tocopherol.

[20] Tocopherols have antioxidant properties which advantageously protect crude oil from oxidation and therefore from rancidity. In addition tocopherols have beneficial effects on health, in particular in prevention against cardiovascular diseases. Crude oil comprising a tocopherol can therefore have beneficial effects on health, in particular in prevention against cardiovascular diseases.

[21] The mass concentration of tocopherol in crude oil can be greater than or equal to 350 ppm, in particular from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2,000 ppm to 3,000 ppm. For the same biological substrate, this mass concentration of tocopherol is greater than the mass concentration of tocopherol in a crude oil produced by processes using anhydrous hexane or 2-methyloxolane.

[22] According to a very particular embodiment, the mass concentration of tocopherol in the crude oil can be greater than or equal to 350 ppm, in particular from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2000 ppm to 3000 ppm, with the exception of the crude oil produced from the biological substrate which is rapeseed, the mass concentration of tocopherol in the crude oil can be from 825 ppm to 10,000 ppm, more particularly from 1500 ppm to 5000 ppm, more particularly still from 2000 ppm to 3000 ppm. [23] Crude oil may also include 2-methyloxolane.

[24] Typically the mass concentration of 2-methyloxolane in the crude oil can be from 0.5 ppm to 500 ppm, in particular from 50 ppm to 300 ppm.

[25] The method according to the invention uses a solvent comprising 2-methyloxolane (CAS No. 96-47-9) and water. Advantageously, 2-methyloxolane is not classified as toxic to the environment and is of biobased origin because its raw material is typically from bagasse of cane sugar or corn cobs. In addition, 2-methyloxolane is non-toxic when ingested for the amounts envisaged. Indeed, a published test of 3 months in ingestion on rats showed a dose in observed adverse effect ("NOAEL - Non Observed Adverse Effect Limit", according to English terminology) of 250 mg / kg body weight / day against 23 mg / kg bodyweight / day for hexane (Parris et al. Regulatory Toxicology and Pharmacology 87 (2017) 54-63 and Opinion of the European Scientific Committee for Food published on June 17, 1994).

[26] According to a particular embodiment, the mass percentage of water in the solvent is from 1% to 15%, more particularly from 4% to 6% during step a) of extraction.

[27] For the purposes of the present invention, "the mass percentage of water in the solvent" corresponds to the mass of water in the solvent divided by the total mass of the solvent at the inlet of the extractor in which the step a) of extraction is carried out. Typically, the mass percentage of water in the solvent can be determined by the Karl Fischer method.

[28] Under certain conditions, in particular in the ranges of mass percentage of water in the solvent indicated above, 2-methyloxolane and water can form homogeneous liquid mixtures.

[29] Indeed an azeotrope comprising 89.4% by weight of 2-methyloxolane and 10.6% by weight of water is formed at 71 ° C. In addition, the water is partially soluble in 2-methyloxolane, the solubility of water in 2-methyloxolane being 4.1% by weight at 20 ° C and 4.6% by weight at 60 ° C. [30] Advantageously, the implementation of the method according to the invention is simpler when the solvent is a homogeneous liquid mixture than when the solvent is a two-phase liquid mixture. This is typically the case for step a) of extraction of the process according to the invention.

[31] For the purposes of the present invention, the term "biological substrate" means a solid material chosen from a plant, an alga, a microorganism and their mixtures, in particular a plant.

[32] When the biological substrate is a vegetable, then the crude oil obtained according to the method according to the invention is called crude vegetable oil.

[33] For example, the vegetable can be an oilseed, a protein crop or a mixture thereof.

[34] As oilseeds, mention may be made of rapeseed, sunflower, soy, peanut, sesame, walnut, almond, cotton, flax or their mixtures, in particular rapeseed, sunflower, soy or their mixtures.

[35] Examples of protein crops are peas, beans, lupine and their mixture.

[36] Typically the biological substrate can be a plant chosen from almond, amaranth, peanut, argan, sea buckthorn, cashew, avocado, oats, borage, safflower , camelina, carrot, cocoa, cashew, hemp, rapeseed, copra, squash, cotton, croton, rose hips, fig, prickly pear, pomegranate, hops, illipe, jojoba, shea, flax, lupine, corn, hazel, walnut, coconut, flax, olive, evening primrose, palm kernel, paprika, pecan, pistachio, pepper, castor, rice, rosebush, sesame, soy, marigold, sunflower, Calophyllum inophyllum, madhuca, Queensland walnut, raspberry, blackcurrant, melon, grape, tomato, baobab, babassu, cranberry, chia, pumpkin, mustard, neem, Nigella Sativa, niger, poppy, Perilla, Plukenetia volubili, pumpkin, annatto, Taramira, l apricot, plum, peach, wheat, par especially soy, rapeseed, sunflower, baobab, corn, peanut, coconut, palm kernel, cotton, especially soy, rapeseed, sunflower, baobab and mixtures thereof. [37] In the case where the biological substrate is a plant, step a) of extraction can be carried out from the whole plant or from one or more parts of the plant, and in particular a part chosen from the root , the stem, the bark, the flower, the seed, the germ, the leaf, the bran, the fruit, the nut, the seeds, the nucleus.

[38] Depending on the plant, the skilled person will know which part to choose.

[39] Typically the organic substrate can be chosen from oat bran, raspberry seeds, blackcurrant seeds, pomegranate seeds, melon seeds, grape seeds, tomato seeds, seeds baobab, babassu seeds, cranberry seeds, chia seeds, corn seeds, cotton seeds, peanut seeds, rapeseed seeds, squash seeds, madhuca seeds, seeds mustard, neem seeds, Nigella Sativa seeds, niger seeds, poppy seeds, Perilla seeds, Plukenetia volubili seeds, pumpkin seeds, annatto seeds, rice bran, soybeans, taramira seeds, sunflower seeds, apricot kernels, plum kernels, peach kernels, Calophyllum inophyllum, walnuts, cashews, macadamia nuts, coconut, pecan, especially soybeans, colz seeds a, sunflower seeds, baobab seeds and their mixtures.

[40] According to a very particular embodiment, the biological substrate is chosen from soybeans, rapeseed, sunflower seeds and their mixtures.

[41] When the biological substrate is an alga, it can be chosen from the genera Arthrospira, Haematococcus, Dunaliella, Chlorella, Nannochloropsis, Schizochytrium, Crypthecodinium, Culindrotheca, Isochrysis, Nannochloris, Nitzchia, Phaeodactylum, Chaetoceros and their mixtures.

[42] When the biological substrate is a microorganism chosen from yeast, mold, bacteria, fungus or a mixture thereof. The yeast can typically be chosen from the genera Cryptococcus, Candida, Lipomyces, Rhodotorula, Saccharomyces, Trichosporon, Yarrowia and their mixtures. [43] Typically, the biological substrate may have undergone, before the extraction step a), a preliminary preparation step. This preparation step can be, for example, a flattening also called flaking to obtain a flake, grinding, mechanical extraction to obtain a flake, pressure, centrifugation, cooking, lyophilization, enzymatic lysis, mechanical lysis , a maceration, a trituration to obtain an oil cake, an ultrasonic treatment, a microwave treatment, a drying or their mixtures or any combination of the said preparations.

[44] Step a) of extraction can typically be carried out in batch ("batch" in English) or continuously.

[45] When step a) of extraction is carried out in a batch, the biological substrate and the solvent are advantageously mixed in an extractor, such as a fixed bed extractor or an extractor with dispersed charge. When stage a) of extraction is carried out in batch the mass ratio biological substrate: solvent can be from 1: 1 to 1: 50, in particular from 1: 2 to 1: 20, more particularly from 1: 5 to 1 : 10.

[46] When step a) of extraction is carried out continuously, the biological substrate arrives continuously in an extractor in a movable bed, such as extractors with movable compartments, movable baskets, conveyor chains or conveyor belt. The solvent is brought into contact with the biological substrate by circulation or percolation, typically against the current of the biological substrate. When step a) of extraction is carried out continuously, the mass ratio of biological substrate: solvent can advantageously be from 1: 0.5 to 1: 5, in particular from 1: 0.75 to 1: 3, more particularly from 1: 1 to 1: 1, 3.

[47] Typically step a) of extraction is advantageously carried out at a temperature of 20 ° C to 100 ° C, in particular from 40 ° C to 80 ° C, more particularly from 55 ° C to 75 ° C. Advantageously, the extraction step a) is facilitated in these temperature ranges because they are compatible with the boiling point of the azeotrope 2-methyloxolane / water which is 71 ° C.

[48] According to one embodiment, the method according to the invention may further comprise a step b) of recovery of the liquid fraction comprising the crude oil and the solvent. [49] Typically, the method according to the invention can comprise, in addition to step a) of extraction, the following steps:

b) recovery of the liquid fraction comprising the crude oil and the solvent, and

c) separation of the crude oil and the solvent from the liquid fraction to recover on the one hand the crude oil and on the other hand the solvent.

[50] Step b) of recovery of the liquid fraction makes it possible to separate the liquid fraction from the solid residue. Typically this step b) can be carried out by filtration.

[51] Step c) of separation can be carried out by liquid / liquid extraction, by steaming, by heating, by distillation or a combination thereof, in particular by steaming, by distillation or combinations thereof, especially by distillation.

[52] Typically step c) of separation can be carried out in batch or continuously.

[53] Steps b) and c) are conventional steps in the processes for producing oils by solid / liquid extraction. Those skilled in the art will be able to adapt the operating conditions of these steps to carry them out.

[54] Typically, the method according to the invention may comprise, after step c), a step d) of recycling in step a) of extraction of all or part of the solvent recovered at the end of the step vs).

[55] Advantageously, recycling the solvent recovered at the end of step c) makes it possible to reduce the economic cost of the raw material of the process according to the invention and therefore to improve its industrial viability.

[56] According to a first variant, all or part of the solvent recovered at the end of step c) undergoes, before step d) of recycling, a step d ') of reducing the mass percentage of water. This first variant is particularly advantageous when the mass percentage of water in the solvent recovered at the end of step c) is greater than 20%, in particular greater than 15%, very particularly greater than 6%. [57] According to a second variant, all or part of the solvent recovered at the end of step c) is directly recycled in step a) of extraction during step d) of recycling. Thus in this second variant, the method does not include, between step c) and step d), step d ') of reducing the mass percentage of water. This second variant is particularly advantageous when the mass percentage of water in the solvent recovered at the end of step c) is from 0.3% to 20%, in particular from 1% to 15%, more particularly from 4% at 6%.

[58] According to one embodiment, step d ') of reducing the mass percentage of water can be carried out by condensation, by decantation, by distillation or their combinations, in particular by condensation, by decantation or their combination, more particularly by decantation followed by distillation.

[59] To facilitate recycling of the solvent, it may be advantageous to form an azeotrope comprising 89.4% by weight of 2-methyloxolane and 10.6% by weight of water at 71 ° C. during condensation. In this case the mass percentage of water in the solvent can be from 1% to 25%, in particular from 5% to 20% and more particularly from 10% to 12% after the condensation. It may also be advantageous to dissolve the water in 2-methyloxolane directly during condensation or possible decantation, the solubility of the water in 2-methyloxolane being 4.1% at 20 ° C. and 4.6% at 60 ° C. In this case, the mass percentage of water in the solvent can be from 0.3% to 20%, in particular from 1% to 15%, more particularly from 4% to 6% after condensation or possible decantation.

[60] A person skilled in the art will know how to adapt the operating conditions for condensation to form the azeotrope and / or dissolve the water in 2-methyloxolane.

[61] A person skilled in the art will also know how to adapt the operating conditions of possible decantation to dissolve the water in 2-methyloxolane.

[62] According to a specific embodiment, step d ') of reducing the mass percentage of water can be carried out by condensation followed by decantation and:

- the mass percentage of water in the solvent after condensation is from 1% to 25%, in particular from 5% to 20% and more particularly from 10% to 12%, and

- The mass percentage of water in the solvent after decantation is from 0.3% to 20%, in particular from 1% to 15%, more particularly from 4% to 6%.

[63] Crude oil can include impurities such as gums, waxes, free fatty acids, pigments, metallic traces, volatile odor compounds and mixtures thereof. The crude oil can therefore undergo a refining step to remove at least one of these impurities from the crude oil and recover a refined oil. Advantageously, this refined oil can be adapted, for example, for food use, for cosmetic use, for pharmaceutical use and / or for technical use.

[64] An embodiment of the invention is a method for producing a refined oil comprising a step of refining the crude oil recovered during step c) of the method for producing a crude oil according to the invention.

[65] The refined oil produced by the process for producing a refined oil according to the invention can comprise a tocopherol. Advantageously, a refined oil comprising a tocopherol can have beneficial effects on health.

[66] The mass concentration of tocopherol in the refined oil may be greater than or equal to 350 ppm, in particular from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2,000 ppm to 3,000 ppm. For the same biological substrate, this mass concentration is greater than the mass concentration of tocopherol in a refined oil produced by processes using anhydrous hexane or 2-methyloxolane.

[67] According to a very particular embodiment, the mass concentration of tocopherol in the refined oil can be greater than or equal to 350 ppm, in particular from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2000 ppm to 3000 ppm, with the exception of the refined oil produced from the biological substrate which is rapeseed, the mass concentration of tocopherol in the crude oil can be from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2,000 ppm to 3,000 ppm.

[68] The refined oil produced by the process for producing a refined oil according to the invention can comprise 2-methyloxolane.

[69] The mass concentration of 2-methyloxolane in the crude oil decreases during the refining stage. Thus the mass concentration of 2-methyloxolane in the refined oil is very low. Typically, the mass concentration of 2-methyloxolane in the refined oil can be less than or equal to 5 ppm, in particular from 0.01 ppm to 3 ppm, more particularly from 0.1 ppm to 1 ppm.

[70] Advantageously, the refined oil produced by the process for producing a refined oil according to the invention does not include hexane. Thus, this refined oil is safer than a refined oil obtained by a conventional process using hexane because, unlike 2-methyloxolane, hexane is neurotoxic and is classified as toxic for reproduction of category 2.

[71] A refined oil with a mass concentration of 2-methyloxolane greater than 5 ppm has degraded organoleptic properties. The refined oil produced by the process for producing a refined oil according to the invention therefore advantageously has satisfactory organoleptic properties.

[72] According to one embodiment, the refining step can be a chemical refining step and / or a physical refining step.

[73] The chemical refining step can comprise at least one of the following substeps:

- degumming to eliminate and recover gums, also called lecithins,

- basic chemical neutralization, in particular with sodium hydroxide, to eliminate free fatty acids,

- discoloration to remove pigments and recover a discolored oil, and - deodorization to eliminate volatile odorous compounds and recover a distillate.

[74] The physical refining step can include at least one of the following substeps:

- degumming to eliminate and recover gums, also called lecithins,

- discoloration to remove pigments and recover a discolored oil, and

- distillation, in particular vacuum distillation with steam injection, to remove volatile odorous compounds and free fatty acids, recovered in the form of distillate.

[75] These chemical refining and physical refining sub-stages are classic sub-stages of the production processes of refined oils by solid / liquid extraction. Those skilled in the art will be able to adapt the operating conditions of these sub-steps to carry them out.

[76] Advantageously, the lecithins recovered during the degumming sub-step and the distillate recovered during the distillation step include polyphenols. They are therefore good for health.

[77] The lecithins recovered during the degumming sub-step can in particular be used as an emulsifier.

[78] The distillate recovered during the distillation step can for example be used in a pharmaceutical, nutraceutical or cosmetic composition. It can also be used as a food supplement.

[79] The method according to the invention also makes it possible to produce a solid residue.

[80] According to a second aspect, the invention relates to a process for producing a solid residue comprising in a step e) recovery of the solid residue obtained during step a) of solid / liquid extraction of the process of production of a crude oil rich in polyphenol according to the invention. [81] For the purposes of the present invention, the term "solid residue" means the defatted solid produced by the process for producing a solid residue according to the invention from the biological substrate. The solid residue is also called oil cake.

[82] The process according to the invention makes it possible to produce a crude oil rich in polyphenol from the biological substrate, the solid residue is therefore depleted in polyphenol.

[83] The mass concentration of polyphenol in the solid residue produced by the process for producing a solid residue according to the invention is less than the mass concentration of polyphenols in a solid residue produced by a conventional process using hexane .

[84] The solid residue produced by the process for producing a solid residue according to the invention comprises a polyphenol and a residual oil, the mass concentration of polyphenol in said solid residue being less than or equal to 3000 ppm, in particular 10 ppm to 1500 ppm, more particularly from 50 ppm to 500 ppm and the mass concentration of residual oil in said solid residue being less than or equal to 5%, in particular from 0.1% to 3%, more particularly from 0.3% at 2%.

[85] Advantageously, having a mass concentration of polyphenols in these value ranges makes it possible to reduce, or even avoid, organoleptic problems, such as the appearance of a dark coloring and / or bitterness, in flours, protein isolates and protein concentrates from these solid residues.

[86] In addition, for the same biological substrate, the solid residue produced by the process for producing a solid residue according to the invention advantageously has a mass concentration of residual oil lower than a solid residue produced by a conventional process using hexane or a process using anhydrous 2-methyloxolane.

[87] For the purposes of the present invention, a "residual oil" included in the solid residue is an oil extracted according to the process described in standard NF EN ISO 734: Feb 2016. [88] Step e) of recovery of the solid residue makes it possible to separate the liquid fraction from the solid residue. Typically this step e) can be carried out by filtration.

[89] The solid residue may comprise solvent for the process according to the invention, in particular 2-methyloxolane. The solid residue can therefore undergo a desolvation step f) after step e) to recover on the one hand the solvent and on the other hand a desolvated solid residue.

[90] Thus another embodiment of the invention is a process for producing a desolvated solid residue comprising a step f) of desolvation of the solid residue recovered during step e) of the process for producing the solid residue according to the invention for recovering on the one hand the solvent and on the other hand the desolvated solid residue.

[91] According to one embodiment, the desolvation step f) can be carried out by heating the solid residue then injecting steam into the heated solid residue, optionally accompanied by placing under vacuum.

[92] For example, the vapor may be a vapor of the solvent according to the invention, a superheated vapor of the solvent according to the invention, a vapor of anhydrous 2-methyloxolane, a superheated vapor of anhydrous 2-methyloxolane, a vapor of water, a superheated vapor of water and their mixtures, in particular a superheated vapor of the solvent according to the invention or a superheated vapor of anhydrous 2-methyloxolane, more particularly a superheated vapor of the solvent according to the invention.

[93] Conventionally, a desolvation step reduces the mass concentration of 2-methyloxolane present in the solid residue. Thus, the desolvated solid residue comprises 2-methyloxolane and the mass concentration of 2-methyloxolane can typically be less than 1000 ppm, in particular from 10 ppm to 500 ppm, very particularly from 100 ppm to 300 ppm.

[94] Typically the mass concentration of residual oil in the desolvated solid residue is less than or equal to 5%, in particular from 0.1% to 3%, more particularly from 0.3% to 2%. [95] According to one embodiment, the solid residue can undergo a pretreatment step before step f) of desolvation such as an additional wetting to adjust the water content in the solid residue. Advantageously, this additional wetting step can facilitate the desolvation of the solid residue. A person skilled in the art will know how to adapt this additional wetting step as a function of the desired water content in the solid residue.

[96] The desolvated solid residue can then be transformed into a by-product intended, for example, for animal feed or human food.

[97] Another embodiment of the invention is therefore a process for producing a by-product comprising a step g) of processing the desolvated solid residue recovered during step f) of the process for producing a solid residue desolvated according to the invention to produce the by-product.

[98] Advantageously, the by-product produced by the process for producing a by-product according to the invention is particularly suitable for feeding animals, in particular cattle. In fact it is degreased, because it comes from the solid residue, and does not disturb the digestion of animals, in particular cattle. In addition, this by-product is safer than a by-product obtained by a conventional process using hexane because, unlike 2-methyloxolane, hexane is neurotoxic and is classified as toxic for reproduction category. 2.

[99] According to one embodiment, the by-product can be chosen from a flour, a protein concentrate, protein isolate, a textured protein and their mixtures.

[100] For the purposes of the present invention, the term "flour" means the by-product from the grinding, milling or spraying of the solid residue in order to obtain a powder.

[101] For the purposes of the present invention, the term "protein concentrate" means a by-product resulting from the treatment of the solid residue by solid extraction. liquid so as to remove the sugars and anti-nutritional factors, so as to obtain a solid fraction containing approximately 60 to 70% of proteins.

[102] For the purposes of the present invention, the term "protein isolate" means a by-product obtained from treatment in an aqueous medium via a succession of steps of the solid residue so as to obtain a solid fraction containing approximately 90% protein.

[103] For the purposes of the present invention, the term "textured protein" means a by-product resulting from the treatment by extrusion of a flour or a protein concentrate.

[104] According to one embodiment, the transformation step g) can be chosen from a grinding step, a solid-liquid extraction step, a protein solubilization step, a protein precipitation step, a step centrifugation, an extrusion step, a protein modification step, a functionalization step of proteins or their mixtures.

[105] A person skilled in the art will know how to choose and adapt the processing step g) according to the by-product he wishes to obtain.

[106] For example, to produce a flour, the processing step g) can be a grinding step.

[107] For example, to produce a protein concentrate, the transformation step g) can be a solid-liquid extraction step using a hydro-alcoholic mixture.

[108] For example, to produce a protein isolate, step g) of transformation can be a succession of steps of solubilization and precipitation of proteins at specific pH so as to selectively remove the sugars and fibers initially present in the solid residue.

[109] For example, to produce a textured protein, the transformation step g) can be an extrusion step from a flour or a protein concentrate. [110] Step g) of transformation can reduce the mass concentration of 2-methyloxolane. Thus, the by-product comprises 2-methyloxolane and the mass concentration of 2-methyloxolane is less than 1000 ppm, in particular less than 500 ppm, very particularly from 0.5 ppm to 50 ppm.

[111] The mass concentration of 2-methyloxolane in the by-product may depend on the by-product.

[112] For example, the mass concentration of 2-methyloxolane in flour can be less than 1000 ppm, in particular less than 500 ppm, very particularly from 5 ppm to 50 ppm.

[113] The mass concentration of 2-methyloxolane in the protein concentrate, in the protein isolate or in the textured protein can typically be less than 30 ppm, in particular from 0.5 ppm to 20 ppm, very particularly from 8 ppm to 12 ppm.

[114] Typically the mass concentration of residual oil in the by-product is less than or equal to 5%, in particular from 0.1% to 3%, more particularly from 0.3% to 2%.

[115] According to one embodiment, the method according to the invention may further comprise a step h) of recycling in step a) of extraction of all or part of the solvent recovered during step f) of desolvation .

[116] Advantageously, recycling the solvent recovered during step f) of desolvation makes it possible to reduce the economic cost of the raw material of the process according to the invention and therefore to improve its industrial viability.

[117] According to a first variant, all or part of the solvent recovered at the end of step f) of desolvation undergoes, before step h) of recycling, a step h ') of reduction of the mass percentage of water. This first variant is particularly advantageous when the mass percentage of water in the solvent recovered at the end of the desolvation step f) is greater than 20%, in particular greater than 15%, very particularly greater than 6%.

[118] According to a second variant, all or part of the solvent recovered at the end of the desolvation step f) is directly recycled in the extraction step a) during from step h) of recycling. Thus in this second variant, the process does not include, between stage f) of desolvation and stage h) of recycling, of stage h ') of reduction of the mass percentage of water. This second variant is particularly advantageous when the mass percentage of water in the solvent recovered at the end of the desolvation step f) is from 0.3% to 20%, in particular from 1% to 15%, more particularly from 4% to 6%.

[119] Step h ') of reducing the mass percentage of water is carried out under the same conditions as step d') of reducing the mass percentage of water.

[120] According to a particular embodiment, all or part of the solvents recovered during steps c) and f) can be mixed and all or part of their mixture can be recycled in step a) of extraction in step i ) recycling.

[121] According to a first variant, all or part of the solvents recovered during steps c) and f) can be mixed and all or part of their mixture can undergo a step i ') of reducing the mass percentage of water before be recycled in step a) of extraction in a step i) of recycling.

[122] According to a second variant, all or part of the solvents recovered during steps c) and f) can be mixed and all or part of their mixture can be directly recycled in step a) of extraction during a step i) recycling. Thus in this second variant, the method does not include, between steps c) and f) and step i), step i ') of reducing the mass percentage of water.

[123] Step i ') of reducing the mass percentage of water is carried out under the same conditions as step d') of reducing the mass percentage of water.

[124] According to a very particular embodiment, the process for producing a crude oil according to the invention comprises the following steps:

a) solid / liquid extraction of the biological substrate with the solvent to obtain on the one hand a liquid fraction comprising the crude oil and the solvent and on the other hand a solid residue,

b) recovery of the liquid fraction comprising the crude oil and the solvent, c) separation of the crude oil and the solvent from the liquid fraction by steam entrainment to recover on the one hand the crude oil and on the other hand the solvent, and

d) recycling in step a) of all or part of the solvent recovered during step c) after a step d) of reduction of the mass percentage of water in all or part of the solvent recovered at the end of the step c) by condensation followed by decantation and:

the mass percentage of water in the solvent after condensation is from 1% to 25%, in particular from 5% to 20% and more particularly from 10% to 12%, and

- The mass percentage of water in the solvent after decantation is from 0.3% to 20%, in particular from 1% to 15%, more particularly from 4% to 6%.

[125] According to a very particular embodiment, the process for producing a desolvated solid residue according to the invention comprises the following steps: a) solid / liquid extraction of the biological substrate with the solvent to obtain on the one hand a fraction liquid comprising the crude oil and the solvent and on the other hand a solid residue,

b) separation of the liquid fraction and the solid residue,

e) recovery of the solid residue,

f) desolvation of the solid residue recovered during step e) to obtain on the one hand the solvent and on the other hand the desolvated solid residue,

h) recycling in step a) of all or part of the solvent recovered during step f).

[126] According to a very particular embodiment, the process for producing a by-product according to the invention comprises the following steps:

a) solid / liquid extraction of the biological substrate with the solvent to obtain on the one hand a liquid fraction comprising the crude oil and the solvent and on the other hand a solid residue,

b) separation of the liquid fraction and the solid residue,

e) recovery of the solid residue, f) desolvation of the solid residue recovered during step e) to obtain on the one hand the solvent and on the other hand the desolvated solid residue,

g) transformation of the desolvated solid residue recovered during step f) to produce the by-product, and

h) recycling in step a) of all or part of the solvent recovered during step f).

[127] As explained above, the method for producing a crude oil according to the invention makes it possible to produce a crude oil advantageously rich in polyphenol and which can comprise 2-methyloxolane.

[128] Thus, the invention also relates to a crude oil rich in polyphenol derived from a biological substrate, said crude oil comprising one or more polyphenols, 2-methyloxolane, and being characterized in that the mass concentration of polyphenol is greater or equal to 100 ppm, in particular said crude oil is capable of being obtained by the process for producing a crude oil according to the invention.

[129] For the purposes of the present invention, the term “crude oil rich in polyphenol” means a crude oil comprising one or more polyphenols and the mass concentration of polyphenol of which is greater than or equal to 100 ppm, in particular from 320 ppm to 2000 ppm, more particularly from 350 ppm to 1500 ppm, more particularly still from 400 ppm to 1200 ppm.

[130] Typically the mass concentration of 2-methyloxolane in the crude oil can be from 0.5 ppm to 500 ppm, in particular from 50 ppm to 300 ppm.

[131] Crude oil may also include a tocopherol.

[132] The mass concentration of tocopherol in the crude oil can be greater than or equal to 350 ppm, in particular from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2,000 ppm to 3,000 ppm. For the same biological substrate, this mass concentration of tocopherol is greater than the mass concentration of tocopherol in a crude oil produced by processes using anhydrous hexane or 2-methyloxolane. [133] According to a very particular embodiment, the mass concentration of tocopherol in the crude oil can be greater than or equal to 350 ppm, in particular from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2000 ppm to 3000 ppm, with the exception of the crude oil produced from the biological substrate which is rapeseed, the mass concentration of tocopherol in the crude oil can be from 825 ppm to 10,000 ppm, more particularly from 1500 ppm to 5000 ppm, more particularly still from 2000 ppm to 3000 ppm.

[134] The biological substrate is as described above in connection with the process for producing a crude oil according to the invention.

[135] Crude oil can advantageously be good for health because it is rich in polyphenol and can include a tocopherol. Advantageously, the crude oil can be adapted, for example, for food use, for cosmetic use, for pharmaceutical use and / or for technical use.

[136] Thus the subject of the present invention is the use of this crude oil for the preparation of a composition such as a food composition, a cosmetic composition and a pharmaceutical composition.

[137] As explained above, the process for producing a refined oil according to an embodiment of the invention advantageously makes it possible to produce a refined oil comprising a tocopherol so that this refined oil can have beneficial effects on the health.

[138] Thus, an object of the invention is a refined oil obtained from a biological substrate comprising a tocopherol, the mass concentration of tocopherol in the refined oil being greater than or equal to 500 ppm, in particular said refined oil is capable of 'be obtained by the process for producing a refined oil according to an embodiment of the invention.

[139] Typically the mass concentration of tocopherol in refined oil can be greater than or equal to 350 ppm, in particular from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2,000 ppm to 3,000 ppm . For the same biological substrate, this concentration mass by tocopherol is greater than the mass concentration by tocopherol in a refined oil produced by processes using anhydrous hexane or 2-methyloxolane.

[140] According to a very particular embodiment, the mass concentration of tocopherol in the refined oil can be greater than or equal to 350 ppm, in particular from 825 ppm to 10,000 ppm, more particularly from 1,500 ppm to 5,000 ppm, more particularly still from 2000 ppm to 3000 ppm, with the exception of the refined oil produced from the biological substrate which is rapeseed, the mass concentration of tocopherol in the crude oil can be from 825 ppm to 10,000 ppm, more particularly from 1500 ppm to 5000 ppm, more particularly still from 2000 ppm to 3000 ppm.

[141] The refined oil of the invention can also comprise 2-methyloxolane. This refined oil is safer than a refined oil produced by a conventional process using hexane because, unlike 2-methyloxolane, hexane is neurotoxic and is classified as toxic for reproduction category 2.

[142] According to one embodiment, the mass concentration of 2-methyloxolane in the refined oil can be less than or equal to 5 ppm, in particular from 0.01 ppm to 3 ppm, more particularly from 0.1 ppm to 1 ppm.

[143] A refined oil whose mass concentration of 2-methyloxolane is greater than 5 ppm has degraded organoleptic properties. The refined oil according to the invention therefore advantageously has satisfactory organoleptic properties.

[144] The biological substrate is as described above in connection with the process for producing a crude oil according to the invention.

[145] Refined oil that includes a tocopherol can be beneficial for health. Advantageously, the refined oil can be adapted, for example, for food use, for cosmetic use, for pharmaceutical use. [146] Thus the subject of the present invention is the use of this refined oil for the preparation of a composition such as a food composition, a cosmetic composition and a pharmaceutical composition.

[147] The process for producing a refined oil according to an embodiment of the invention also makes it possible to produce lecithins and / or a distillate comprising a polyphenol.

[148] Thus an object of the invention is lecithins from a biological substrate comprising a polyphenol, in particular said lecithins are capable of being obtained by the process for the production of a refined oil according to an embodiment of the 'invention.

[149] The biological substrate is as described above in connection with the process for producing a crude oil according to the invention.

[150] Advantageously, the lecithins include polyphenols. They are therefore good for health.

[151] These lecithins can in particular be used as an emulsifier.

[152] Thus the subject of the present invention is the use of these lecithins as an emulsifier.

[153] An object of the invention is a distillate from a biological substrate comprising a polyphenol, in particular said distillate is capable of being obtained by the process for producing a refined oil according to one embodiment of the invention.

[154] The biological substrate is as described above in connection with the process for producing a crude oil according to the invention.

[155] The distillate can for example be used in a pharmaceutical, nutraceutical or cosmetic composition. It can also be used as a food supplement.

[156] Thus the subject of the present invention is the use of this distillate for the preparation of a composition such as a pharmaceutical composition, a nutraceutical composition or cosmetic composition. The present invention relates to the use of this distillate as a food supplement.

[157] As explained above, the method for producing a solid residue according to an embodiment of the invention advantageously makes it possible to produce a solid residue depleted in polyphenols and comprising a low mass concentration of residual oil.

[158] Thus an embodiment of the invention is a solid residue derived from a biological substrate comprising a polyphenol and a residual oil, the mass concentration of polyphenol of which is less than or equal to 3000 ppm, in particular from 10 to 1500 ppm, more particularly from 50 to 500 ppm and whose mass concentration in residual oil is less than or equal to 5%, in particular from 0.1% to 3%, more particularly from 0.3% to 2%, in particular said solid residue is capable of being obtained by the process for producing a solid residue according to an embodiment of the invention.

[159] Advantageously, having a mass concentration of polyphenols in these value ranges makes it possible to reduce, or even avoid, organoleptic problems, such as the appearance of a dark coloring and / or bitterness, in flours, protein isolates and protein concentrates from these solid residues.

[160] The biological substrate is as described above in connection with the process for producing a crude oil according to the invention.

[161] The residual oil included in the solid residue is as defined above in connection with the process for producing a solid residue.

[162] According to one embodiment, the solid residue can also comprise 2-methyloxolane.

[163] According to one embodiment, the solid residue does not include hexane.

[164] The solid residue can then be transformed into a by-product intended, for example, for animal feed or human food. [165] The present invention also relates to the use of this solid residue for the preparation of an animal or human food composition.

[166] Advantageously, it is possible to produce a desolvated solid residue with the method for producing a desolvated solid residue described above.

[167] Thus, one embodiment of the invention is a desolvated solid residue derived from a biological substrate comprising 2-methyloxolane and whose mass concentration of 2-methyloxolane is less than 1000 ppm, in particular from 10 ppm to 500 ppm, very particularly from 100 ppm to 300 ppm, in particular said desolvated solid residue is capable of being obtained by the process for the production of a desolvated solid residue according to an embodiment of the invention.

[168] Typically the mass concentration of residual oil in the desolvated solid residue is less than or equal to 5%, in particular from 0.1% to 3%, more particularly from 0.3% to 2%.

[169] According to one embodiment, the desolvated solid residue does not include hexane.

[170] The biological substrate is as described above in connection with the process for producing a crude oil according to the invention.

[171] The desolvated solid residue can then be transformed into a by-product intended, for example, for animal feed or human food.

[172] Another subject of the present invention is the use of this desolvated solid residue for the preparation of an animal or human food composition.

[173] Advantageously, it is also possible to produce a by-product which is particularly suitable for food, in particular animal food, with the process for producing a by-product described above.

[174] Thus the invention also relates to a by-product derived from a biological substrate comprising 2-methyloxolane and whose mass concentration in 2-methyloxolane is less than 1000 ppm, in particular less than 500 ppm, very particularly from 0.5 ppm to 50 ppm, in particular said by-product is capable of being obtained by the process for producing a by-product according to one embodiment of the invention.

[175] According to one embodiment, the by-product can be chosen from a flour, a protein concentrate, protein isolate, a textured protein, and their mixtures.

[176] The mass concentration of 2-methyloxolane in the by-product may depend on the by-product.

[177] For example, the mass concentration of 2-methyloxolane in flour can be less than 1000 ppm, in particular less than 500 ppm, very particularly from 5 ppm to 50 ppm.

[178] The mass concentration of 2-methyloxolane in the protein concentrate, in the protein isolate or in the textured protein can typically be less than 30 ppm, in particular from 5 ppm to 20 ppm, very particularly from 8 ppm to 12 ppm .

[179] Typically the mass concentration of residual oil in the by-product is less than or equal to 5%, in particular from 0.1% to 3%, more particularly from 0.3% to 2%.

[180] According to one embodiment, the by-product does not include hexane.

[181] The biological substrate is as described above in connection with the process for producing a crude oil according to the invention.

[182] The by-product being particularly suitable for food, in particular animal food, the present invention also relates to the use of this by-product for the preparation of a food composition, in particular a food composition. animal.

[183] Typically, the mass concentration of polyphenols in crude oil, in solid residue, in lecithins and in distillate is determined according to the method known as "Folin Ciocalteu" described by Slinkard and Singleton in publication "Total Phenol Analysis: Automation and Comparison with Manual Methods" published in the journal "American Journal of Enology and Viticulture 28, no 1 (January 1, 1977): 49-55". Those skilled in the art know how to adapt this method to the sample analyzed and for its implementation on a 96-well microplate reader (FLUOstar Omega, BMG LABTECH, France).

[184] Typically, the mass concentration of tocopherols in crude oil and in refined oil is determined according to standard NF EN ISO 9936: June 2016.

[185] The mass concentration of 2-methyloxolane in crude oil, refined oil, solid residue, desolvated solid residue, flour, textured protein, protein concentrate and protein isolate is measured using using a classical analytical chemistry technique called “GC-Headspace” (gas chromatography with sampling of the head space). This analytical technique is known to those skilled in the art as being suitable for the analysis of volatile compounds contained in an oily matrix and in a solid matrix. The operating conditions depend on the sample and the person skilled in the art will know how to adapt them to this sample.

[186] For example, for a protein isolate, the operating conditions are as follows: 0.50 ± 0.01 g of sample to be analyzed are weighed in a 20 mL bottle, specially designed for the "HeadSpace" technique (23 x 77 mm), then 7.0 mL of ultrapure water and 1.0 mL of DMF (N, N-dimethylformamide) are added to the bottle. The bottle is then closed hermetically using a cap fitted with a suitable septum. The bottle is then shaken vigorously for 30 seconds before analysis. The operating conditions of the head space removal and injection stage are as follows: instrument = HeadSpace module 7697A; desorption = 20 min at 100 ° C; transfer line temperature = 160 ° C; injection loop temperature = 130 ° C; injection volume = 1 pL; helium pressure = 12 PSI. The operating conditions for the chromatographic separation step are as follows: instrument = GC 7890A (Agilent); column = DB624 - 60 m - 320 pm - 1, 8 pm; liner = Agilent 5190-6168 (Ultralnert, splitless, straight, 2 mm id); injector temperature = 250 ° C; nature and flow rate of the carrier gas = He, 1.3 mL / min; Oven temperature profile = 60 ° C (3 min) - 5 ° C / min ramp up to 70 ° C - ramp at 10 ° C / min up to 220 ° C - 220 ° C (2 min). The operating conditions of the detection step are as follows: type of detector = mass spectrometer (MS); ionization = El, SIM mode (ions 56, 71 & 86 Da) for identification and total ion current for quantification; source temperature = 230 ° C; temperature of the quadrupole = 150 ° C. All reagents, solvents and materials used are of suitable analytical quality.

The mass concentration of 2-methyloxolane in the analyzed sample of protein isolate is determined using a calibration curve performed over a range of mass concentrations of suitable 2-methyloxolane.

[187] For the solid residue, the operating conditions are as follows: the solid residue is first ground using a ZM 200 centrifugal mill (Retsch GmbH) equipped with a 0.2 mm sieve. Then 0.5 ± 0.01 g of the ground sample to be analyzed is weighed in a 20 mL bottle, specially designed for the "HeadSpace" technique (23 x 77 mm). Then 2.0 mL of water is added precisely to the bottle before closing it tightly with a cap fitted with a septum. The bottle is left to stand for at least 5 min before analysis. The operating conditions of the head space removal and injection stage are as follows: instrument = "Turbomatrix HS40" (Perkin Elmer); desorption = 60 min at 80 ° C; transfer line temperature = 120 ° C; needle temperature = 1 10 ° C; injection volume = 0.2 mL; hydrogen pressure = 20 PSI. The operating conditions of the chromatographic separation step are as follows: instrument = Clarus 500 (Perkin Elmer); column = DB1 - 30 m - 0.32 mm - 3.0 pm; injector temperature = 1 10 ° C; oven temperature = 40 ° C (7 min, isothermal). The operating conditions of the detection step are as follows: type of detector = flame ionization detector (FID); detector temperature = 250 ° C; hydrogen pressure = 20 PSI. All reagents, solvents and materials used are of suitable analytical quality.

A calibration line drawn beforehand makes it possible to determine the mass concentration of 2-methyloxolane in the sample of solid residue analyzed, according to the following formula: C, = k, ^* A ,, where C, is the concentration mass in 2-methyloxolane, A, the area of the peak corresponding to 2-methyloxolane and k, the slope of the calibration line for 2-methyloxolane.

[188] The mass concentration of residual oil in the solid residue is determined according to standard NF EN ISO 734: Feb 2016.

Examples

[189] In the examples below, the method for determining the mass concentration of polyphenols in crude oils is the so-called “Folin Ciocalteu” method, described by Slinkard and Singleton in the publication “Total Phenol Analysis: Automation and Comparison with Manual Methods ”published in the journal“ American Journal of Enology and Viticulture 28, no 1 (January 1, 1977): 49-55 ”, with the modifications described below to implement it on a 96-well microplate reader (FLUOstar Omega , BMG LABTECFI, France). All the reagents and solvents used are of suitable analytical quality.

The polyphenols contained in the crude oil are previously extracted by the following method: 1 g of oil is diluted in 1 ml of hexane, then the solution is extracted by 3 successive extractions with 3 ml of a methanol / water mixture ( 60% vol / 40% vol). After each extraction, the 2 phases are agitated and then separated by centrifugation (10,000 rpm-1/10 min / 20 ° C) and the liquid alcoholic phase is collected (any deposits, precipitates or potentially formed solid particles are not not taken). The 3 hydroalcoholic phases are combined and then washed with 1 ml of hexane. The phases are separated by centrifugation then the hydro-alcoholic phase is transferred to a 10 ml volumetric flask, and the volume is made up to the mark with the methanol / water mixture (60% vol / 40% vol).

Using a micropipette, 20 μL of this solution are introduced into a well of the microplate to which 80 μL of aqueous solution of Na 2 CO 3 (anhydrous, Acros Organics) at 75 g / L are added. The same mixture is repeated in 7 other wells, ie 8 wells in total for 1 sample of crude oil analyzed. Then 100 μl of Folin-Ciocalteu reagent (Panreac AppliChem, ref. 251567.1609), previously diluted 1/10 (v / v) in distilled water, are added to each well in an automated manner by the automatic injector of the reader microplate. The The absorbance of each well is read by the UV-Visible detector of the microplate reader at 750 nm at 25 ° C after 1 h of agitation in the dark in the device. In parallel, a standard curve was established using 8 aqueous solutions of gallic acid (Sigma-Aldrich) in a concentration range from 0 to 100 mg / L of gallic acid, according to the same protocol. analysis defined for the samples, the difference being that the solution comprising the phase and the methanol / water mixture is replaced by one of the 8 aqueous solutions of gallic acid.

In a second step, in order to eliminate the non-polyphenolic compounds present in the oil and which can react with the Folin-Ciocalteu reagent (reducing sugars, proteins, etc.), 5 mL of the polyphenol solution obtained previously from crude oil are acidified to pH = 3.5 with a 0.1N HCl solution. Then to this mixture are added 5 mL of distilled water and 1 g of polyvinylpolypyrrolidone (PVPP, particle size ^“ 1 10 pm, Sigma-Aldrich) in order to capture the polyphenols. The mixture is mechanically stirred for 10 min at 25 ° C before being centrifuged at 10,000 rpm for 10 min at 20 ° C. The supernatant is then removed and then filtered using a syringe filter (0.25 μm) before being analyzed according to the same protocol as the solution not treated with PVPP. The absorbance value of this solution treated with PVPP will be used as "blank", it will be subtracted from the value obtained previously.

The absorbance value thus obtained, average from the absorbance measurement of 8 wells, from which the contribution of non-polyphenolic compounds is subtracted, is used to calculate the mass concentration of polyphenols in the sample, from the equation of the calibration line. Considering the mass of oil initially used for the analysis, the mass concentration of polyphenols in the sample is expressed in pg of gallic acid (EAG, abbreviated) / g of crude oil, or equivalent, in ppm.

[190] In the examples below, the method for determining the mass concentration of tocopherols in crude oils and in refined oils is standard NF EN ISO 9936: June 201 6 .. [191] In the examples below, the method for determining the mass concentration of 2-methyloxolane in the refined oil is implemented using the conventional technique of analytical chemistry known as "GC-HeadSpace" (Chromatography in gas phase with removal of the headspace), under the conditions described below. This analytical technique is known to those skilled in the art as being suitable for the analysis of volatile compounds contained in an oily matrix.

All reagents, solvents and materials used are of suitable analytical quality. First, 5 ± 0.01 g of refined oil to be analyzed are weighed in a 20 mL bottle, specially designed for the "HeadSpace" technique (23 x 77 mm). Then 15 ml of a standard solution (heptane 40% v / v in octane) are added precisely to the 20 ml bottle before closing it hermetically using a cap fitted with a septum . The 20 mL bottle is shaken vigorously (vortex) for 5 min before analysis. The operating conditions of the head space removal and injection stage are as follows: instrument = "Turbomatrix HS40" (Perkin Elmer); desorption = 60 min at 80 ° C; transfer line temperature = 120 ° C; needle temperature = 1 10 ° C; injection volume = 0.2 mL; hydrogen pressure = 20 PSI. The operating conditions of the chromatographic separation step are as follows: instrument = Clarus 500 (Perkin Elmer); column = DB1 - 30 m - 0.32 mm - 3.0 pm; injector temperature = 150 ° C; oven temperature = 40 ° C (3 min) then ramps from 10 ° C / min to 1 10 ° C (0 min). The operating conditions of the detection step are as follows: type of detector = flame ionization detector (FID); detector temperature = 250 ° C; hydrogen pressure = 1 1 PSI.

The mass concentration of 2-methyloxolane in the refined oil sample analyzed is determined according to the following formula: OM _Q O _C = a * (A _Me ox / A _hep tane) where Cwieox is the mass concentration of 2-methyloxolane, A _Me ox the area of the peak corresponding to 2-methyloxolane, A _hep tane the area of the peak corresponding to heptane (internal standard), and the slope of the calibration line for 2-methyloxolane established previously.

The calibration curve has been drawn, according to the classical principles of chemistry analytical, by adding known quantities of a solution containing 2-methyloxolane and heptane, in a refined oil devoid of 2-methyloxolane, so as to obtain mass concentrations in oil of 0.51; 1.02; 2.05; 5.12 and 10.25 pg / g.

[192] Example 1 according to the invention: the solvent comprises 2-methyloxolane and water

[193] Example 1 -1: the substrate is the soybean

[194] The extraction of crude soybean oil was carried out using an automatic Soxhlet extraction system (Extraction System B-81 1, Büchi), from shelled soybeans ( Supplier: OLEAD, variety: ES PALLADOR, harvest: France, 2017, water content: 8.5% +/- 0.3). The solvent used is a mixture of 2-methyloxolane (stabilized with BHT, Sigma Aldrich) containing 4.5 g of distilled water per 100 g of solvent.

[195] About 50 g of soybeans are ground using a knife mill to obtain particles less than 1 mm in size.

About 30 g of the powder obtained are weighed and placed in a glass extraction cartridge adapted to the device (Büchi). The cartridge is then introduced and fixed in the Soxhlet chamber according to the instructions defined in the user manual of the device.

Then, 170 mL of solvent are introduced into the 250 mL receptacle cup provided for this purpose. Then the appliance is adjusted so as to operate according to the "Soxhlet Standard" mode, without rinsing or drying, with a number of cycles fixed at 20 and a heating power fixed at 10.

Finally, the level detector is placed approximately 1 cm above the "high" level of the plant substrate and the refrigerant is supplied with cold water (8 ° C).

The solvent is then brought to a boil using the built-in hot plate. After the 20 cycles are complete, all the solvent containing the extracted oil is collected in the receiving cup, while the solid residue remains inside the extraction cartridge.

The solvent containing the extracted oil is left to cool for approximately 20 min at room temperature before being transferred to a 250 ml flask. The solvent is then evaporated using a rotary evaporator under reduced pressure (150 rpm; 50 ° C; 180 mbar, then 1 mbar to finish).

The crude oil thus obtained is weighed and then cooled under a light flow of nitrogen for 10 minutes before being transferred to a sealed tube and then stored in the freezer at -20 ° C while awaiting analyzes.

[196] The mass concentration of polyphenols in crude oil is presented in Table 1.

[197] Example 1 -2: the substrate is rapeseed

[198] The operating protocol is the same as in Example 1-1, the difference being that the starting substrate is the whole rapeseed (supplier: OLEAD; provenance: Gironde (France); harvest: 2016; water: 5.2% +/- 0.15) and that the Soxhlet extraction cartridges used are made of cellulose and not glass.

[199] The mass concentration of polyphenols in crude oil is presented in Table 1.

[200] Example 1 -3: the substrate is the grain of corn

[201] The operating protocol is the same as in Example 1 -1, the differences being that the starting substrate is the grain of corn (supplier: University of Avignon; provenance: France; water content = 7.25 %), that the Soxhlet extraction cartridges used are made of cellulose and not glass, that the level of the sensor has been fixed so that the volume of the extraction chamber is approximately 175 mL, that the duration of extraction was set at 1 h without setting the number of cycles and the heating power was set at 12.

[202] The mass concentration of polyphenols in crude oil is presented in Table 1.

[203] Example 1 -4: the substrate is cotton seed

[204] The operating protocol is the same as in Example 1 -3, the differences being that the starting substrate is cotton seed (supplier: University of Avignon; provenance: Turkey; water content = 7.99 %) and that seeds have been previously crushed and sieved so as to recover only the almond, without the cotton fiber.

[205] The mass concentration of polyphenols in crude oil is presented in Table 1.

Comparative Example 2: the solvent is hexane or the anhydrous 2-methyloxolane

[207] Comparative Example 2-1: Hexane

[208] The operating protocol and the biological substrates are the same as in Examples 1 -1 and 1 -4, the difference being that the solvent is hexane.

[209] The mass concentration of polyphenols is presented in the Table

1.

[210] Comparative Example 2-2: Anhydrous 2-methyloxolane

[211] The operating protocol and the biological substrates are the same as in Examples 1 -1 and 1 -4, the difference being that the solvent is anhydrous 2-methyloxolane.

[212] The mass concentration of polyphenols is presented in Table 1.

[213] The results of Table 1 show that the process according to the invention using a solvent comprising 2-methyloxolane and water makes it possible to produce an oil richer in polyphenol than the oil obtained by hexane (reference solvent) and with anhydrous 2-methyloxolane.

[214] [Table 1]

[215] Example 3: Impact of the mass concentration of water in the solvent

[216] In this example, a crude oil is obtained by solid / liquid extraction from three different biological substrates with different solvents comprising 2-methyloxolane (stabilized with BHT, Sigma Aldrich) and distilled water, the mass percentage in water in each solvent being 1%, 4.5%, 10% or 20%.

[217] The three biological substrates are:

- soybeans (supplier: OLEAD, variety: ES PALLADOR, harvest: France, 2017; water content: 8.5% +/- 0.3),

- rapeseed (supplier: OLEAD; provenance: Gironde (France); harvest: 2016; water content: 5.2% +/- 0.15), and

- sunflower seeds (provenance: Spain; supplier: L'île aux épices; lot LPR22-1017; water content: 2.54% +/- 0.12).

The edible oils extracted from these three oil seeds are among the most produced in the world excluding palm oil.

[218] In this Example 3, the solid / liquid extraction is carried out by the so-called reflux maceration extraction method because this method is more suitable for the two-phase solvent than the "Soxhlet" extraction method.

[219] Example 3-1 according to the invention: the substrate is soybean

[220] In this example 3-1, the biological substrate is soybean. [221] About 50 g of shelled soybeans are ground using a knife mill to obtain particles less than 1 mm in size. 30 g of the powder obtained are weighed and introduced into a 250 ml glass flask surmounted by a condenser so as to be able to carry out a reflux extraction.

Then, 170 mL of each solvent is introduced into the flask and the contents of the flask are brought to reflux using a flask heater.

The duration of the extraction is fixed at 2 hours from the first sign of reflux. After 2 hours, the heating is stopped and the mixture is allowed to cool to room temperature for 20 min.

Thereafter, the contents of the flask are filtered on a cotton bed so as to separate on one side the solid residue from the solvent containing the oil, collected in a new 250 ml flask. The solvent is then evaporated using a rotary evaporator under reduced pressure (150 rpm; 50 ° C; 180 mbar, then 1 mbar to finish). The crude oil thus obtained is weighed and then the traces of residual solvent are removed by a light flow of nitrogen for 10 minutes. The oil is transferred to a sealed tube and then stored in the freezer at -20 ° C while waiting for the analyzes.

[222] For each solvent exemplified, the mass concentration of polyphenols is presented in Table 2.

[223] Example 3-2 according to the invention: the substrate is rapeseed

[224] The procedure of this Example 3-2 according to the invention is identical to Example 3-1 according to the invention with the exception of the substrate which is rapeseed.

[225] For each solvent exemplified, the mass concentration of polyphenols is presented in Table 3.

[226] Example 3-3 according to the invention: the substrate is the sunflower seed

[227] The procedure of this Example 3-3 according to the invention is identical to Example 3-1 according to the invention with the exception of the substrate which is the husked sunflower seed.

[228] For each solvent exemplified, the mass concentration of polyphenols is presented in Table 4. [229] Comparative Example 3-4: Hexane

[230] The operating protocol and the biological substrates are the same as in Examples 3-1 to 3-3, the difference being that the solvent is hexane.

[231] For each biological substrate, the mass concentration of polyphenols is presented in Tables 2, 3 and 4.

[232] Comparative Example 3-5: Anhydrous 2-methyloxolane

[233] The operating protocol and the biological substrates are the same as in Examples 3-1 to 3-3, the difference being that the solvent is anhydrous 2-methyloxolane. [234] For each biological substrate, the mass concentration of polyphenols is presented in Tables 2, 3 and 4.

[235] [Table 2]

The substrate is the soybean

MeTHF stands for 2-methyloxolane

[236] [Table 3]

The substrate is rapeseed

MeTHF stands for 2-methyloxolane

[237] [Table 4]

The substrate is the sunflower seed

MeTHF stands for 2-methyloxolane

[238] The results of Tables 2 to 4 show that the process for producing a crude oil according to the invention using a solvent comprising 2-methyloxolane and from 1% to 20% of water makes it possible to produce a oil richer in polyphenols than the oil obtained by hexane (reference solvent) and by anhydrous 2-methyloxolane.

[239] The mass concentrations of total polyphenols obtained by the so-called reflux maceration extraction method (Examples 3 and 4) are logically lower than those obtained by the “Soxhlet” extraction method (Examples 1 and 2). In fact, unlike the "Soxhlet" extraction method, the solvent used in the so-called reflux maceration extraction method gradually concentrates on the extract, thus limiting the extraction of the oil and polyphenols, according to the laws of material transfer known to those skilled in the art.

[240] Example 4: Production of a defatted meal

[241] Example 4-1: the substrate is soy flakes and the solvent comprises 2-methyloxolane and 4.5% water.

[242] Solid / liquid extraction was carried out using an automatic extraction system (Extraction System B-81 1, Büchi), from soy flakes (supplier: OLEAD, harvest: France, 2017, water content: 9.96% +/- 0.20%, oil content = 19.19% +/- 0.20%, thickness "1 mm). The solvent used is a mixture of 2-methyloxolane (stabilized with BHT, Sigma Aldrich) containing 4.5 g of distilled water per 100 g of solvent.

[243] About 15 g of soy flakes are weighed and placed in a cellulose extraction cartridge suitable for the device (Büchi). The cartridge is then introduced into the extraction chamber, according to the instructions defined in the device's user manual. The height of the level detector is adjusted so that the maximum volume of solvent in the extraction chamber is approximately equal to 220 mL.

[244] Next, 170 mL of solvent are introduced into the 250 mL receptacle cup provided for this purpose. Then, the device is configured to operate according to the “Soxhlet Standard” mode, without rinsing or drying, with a duration fixed at 1 hour and a heating power equal to 12 so as to guarantee a number of filling cycles- emptying of the extraction chamber equal to 7 ± 1 per hour. The condensers are supplied with tap water, with a flow sufficient to guarantee condensation of the solvent vapors that will be generated.

[245] The solvent is then brought to a boil using the built-in hot plate. After 60 min of extraction, all the solvent containing the extracted oil is collected in the receiving cup, while the defatted cake remains inside the cartridge. The content of the receiving cup is then transferred to a suitable flask and the mixture is desolvated according to the conditions mentioned in the previous examples. The cartridge containing the defatted oil cake is collected and then placed in a ventilated desiccator (Biosec type, brand = TauRo) operating at a temperature of around 45 ° C for at least 10 hours and in any event until that the solvent odor is no longer perceptible. Once desolvated, the mass concentration of residual oil in the defatted cake is determined according to standard NF EN ISO 734: Feb 2016, implemented using the automatic extraction system (Extraction System B-811, Büchi) according to the "Continuous Extraction" mode corresponding to the so-called "Twisselmann" method, with a slight modification because the material to be analyzed is finely ground using a knife mill instead of a micro ball mill.

[246] The mass concentration of residual oil obtained after extraction with 2-methyloxolane + water is presented in Table 5.

[247] Example 4-2: the substrate is rapeseed and the solvent comprises 2-methyloxolane and 4.5% of water.

[248] Solid / liquid extraction was carried out according to the same protocol as in Example 4-1, with the difference that the substrate is rapeseed scales (supplier: OLEAD, harvest: France, 2017, content in water: 8.21% +/- 0.13%, oil content = 23.03% +/- 0.07%, particle size <3 mm) and that the extraction time is 90 min.

[249] The mass concentration of residual oil obtained after extraction with 2-methyloxolane + water is presented in Table 5. [250] Example 4-3: the substrate is sunflower scales and the solvent comprises 2-methyloxolane and 4.5% water.

[251] Solid / liquid extraction was carried out according to the same protocol as in Example 4-1, with the difference that the substrate is sunflower scales (supplier: OLEAD,, harvest: France, 2017, water content: 5.38% +/- 0.24%, oil content = 30.81% +/- 2.59%, particle size <10 mm) and that the extraction time is 90 min .

[252] The mass concentration of residual oil obtained after extraction with 2-methyloxolane + water is presented in Table 5.

[253] Comparative Example 4-4: Hexane

[254] The operating protocol and the biological substrates are the same as in Examples 4-1 to 4-3, the differences being that the solvent is hexane and that the heating power is fixed at 9 so as to guarantee a number of filling-emptying cycles of the extraction chamber equal to 7 ± 1 per hour.

[255] For each biological substrate, the mass concentrations of residual oil obtained after extraction with hexane are presented in Table 5.

[256] Comparative Example 4-5: Anhydrous 2-methyloxolane

[257] The operating protocol and the biological substrates are the same as in Examples 4-1 to 4-3, the differences being that the solvent is anhydrous 2-methyloxolane and that the heating power is fixed at 12 so as to guarantee a number of filling-emptying cycles of the extraction chamber equal to 7 ± 1 per hour.

[258] For each biological substrate, the mass concentrations of residual oil obtained after extraction with anhydrous 2-methyloxolane are presented in Table 5. [259] [Table 5]

[260] The results of Table 5 demonstrate that the process for producing a solid residue according to the invention using a solvent comprising 2-methyloxolane and 4.5% of water makes it possible to produce a solid residue comprising less residual oil than the solid residue obtained by hexane (reference solvent) and by anhydrous 2-methyloxolane.

[261] This is particularly advantageous because it makes it possible to limit the oil losses, to facilitate the elimination of the residual solvent in the solid residue, to increase the concentration of proteins in the solid residue, to improve its stability during storage and facilitate its digestion by animals, especially cattle.

[262] Example 5: Pilot scale test with soya

[263] Example 5-1 according to the invention: the substrate is soybean and the solvent is a mixture of 2-methyloxolane and water.

[264] The extraction of crude soybean oil was carried out on a pilot scale in a 480 L filter drier ("Guedu" type). The soybeans (Supplier: OLEAD, harvest: France, 2017, water content: 12.2%) were prepared for extraction by conventional shelling and flaking steps so as to reduce the size of the particles and increase the l accessibility of the extraction solvent. Then about 60 kg of soy flakes were introduced into the filter drier, then extracted by three successive passes of 2-methyloxolane (supplier = Pennakem LLC; not stabilized). On each pass, the water content is different, respectively 1.44%; 2.85% then 4.76%.

The extraction temperature was on average 57 +/- 6 ° C, the solvent / solid mass ratio was set at 1.8 kg / kg and the extraction time at 15 min per passage, ie 3 x 15 min in total.

The solvent containing the extracted oil (mixture noted misella) was collected by filtration after each pass, then concentrated using a distillation column (80-85 ° C, under reduced pressure) and finally desolvated with using a rotary evaporator (Hei-VAP Advantage, Heidolph, Germany) under vacuum and at 60 ° C until the absence of solvent condensation, then 1 hour at 90 ° C. The desolvated extract is then centrifuged (4000 rpm / 4 min) in order to remove a solid fraction extracted by the 2-methyloxolane and water mixture but which has become insoluble in an oily medium. The crude oil obtained is then stored at -20 ° C before refining or analysis.

[265] Once the extraction is complete, the solid residue (degreased and filtered flakes) is desolvated in the same tank ("Guedu" filter dryer). The tank is connected to a vacuum generator and then heated to 55 ° C for 140 min, with the injection of a nitrogen stream (14-18 L / min) to help the desolvation. Finally, the solid residue is discharged, spread on a plate and exposed to ambient air for 1 day in order to remove the traces of residual solvent.

[266] The crude oil obtained then undergoes a conventional chemical refining step comprising a degumming step at neutral pH then at acidic pH, followed by a neutralization step, a bleaching step then a deodorization step to obtain an oil refined.

[267] The mass concentration of polyphenols and tocopherols in crude oil, and the mass concentration of tocopherols and 2-methyloxolane in refined oil are presented in Table 6.

[268] Comparative Example 5-2: The substrate is soybean and the solvent is anhydrous 2-methyloxolane.

[269] The extraction operating protocol is identical to Example 5-1 according to the invention with the exception of the following points: the solvent is 2-methyloxolane anhydrous (not stabilized), the solvent / solid ratio is 2.9 kg / kg, the extraction temperature is 53 +/- 5 ° C.

[270] The mass concentration of polyphenols and tocopherols in crude oil, and the mass concentration of tocopherols and 2-methyloxolane in refined oil are presented in Table 6.

[271] Comparative Example 5-3: The substrate is soybean and the solvent is hexane.

[272] The extraction operating protocol is identical to Example 5-1 according to the invention with the exception of the following points: the solvent is hexane (extraction quality), the solvent / solid ratio is 2.2 kg / kg, the extraction temperature is 52 +/- 3 ° C.

[273] The mass concentration of polyphenols and tocopherols in crude oil, and the mass concentration of tocopherols in refined oil are presented in Table 6. [274] [Table 6]

[275] The results of Table 6 show that the process for producing a crude oil according to the invention using a solvent comprising 2-methyloxolane and from 1.44% to 4.76% of water allows to produce a crude oil richer in polyphenols than the crude oil obtained by hexane (reference solvent) and by anhydrous 2-methyloxolane.

[276] The results of Table 6 also demonstrate that the process for producing a crude oil according to the invention using a solvent comprising 2-methyloxolane and from 1.44% to 4.76% of water produces a refined oil richer in tocopherols than the refined oil obtained by hexane (reference solvent) and by anhydrous 2-methyloxolane.

Claims

Claims [Claim 1] Method for producing a crude oil rich in polyphenol from a biological substrate comprising a step of: a) solid / liquid extraction of the biological substrate with a solvent to obtain on the one hand a liquid fraction comprising the crude oil and the solvent and on the other hand a solid residue, characterized in that, the solvent comprises 2-methyloxolane and water, and the mass percentage of water in the solvent during step a) of extraction is from 0.3% to 20%. [Claim 2] Process according to claim 1, in which the biological substrate is chosen from a plant, an alga, a microorganism and their mixtures. [Claim 3] A method according to claim 2 wherein the biological substrate is a plant chosen from almond, amaranth, peanut, argan, sea buckthorn, cashew, avocado, oats , borage, safflower, camelina, carrot, cocoa, cashew, hemp, rapeseed, copra, squash, cotton, croton, rose hips, fig, prickly pear, pomegranate , hops, rillipé, jojoba, shea, flax, lupine, corn, hazel, walnut, coconut, flax, olive, evening primrose, palm kernel, paprika, pistachio , pepper, castor, rice, rosebush, sesame, soy, marigold, sunflower, Calophyllum inophyllum, madhuca, Queensland walnut, raspberry, blackcurrant, melon, grape, tomato, baobab, babassu, cranberry, chia, pumpkin, mustard, neem, Nigella Sativa, niger, poppy, pecan, Perilla, Plukenetia volubili, pumpkin, annatto, Taramira, the shelter cot, plum, peach, wheat. [Claim 4] A method according to any one of claims 1 to 3 wherein step a) of extraction is carried out at a temperature of 20 ° C to 100 ° C. [Claim 5] Method according to any one of claims 1 to 4, comprising, in addition to step a) of extraction, the following steps: b) recovery of the liquid fraction comprising the crude oil and the solvent, and c) separation of the crude oil and the solvent from the liquid fraction to recover on the one hand the crude oil and on the other hand the solvent. [Claim 6] Method for producing a refined oil comprising a step for refining the crude oil recovered during step c) of the process for producing a crude oil as defined in claim 5. [Claim 7] A method for producing a solid residue comprising a step e) of recovering the solid residue obtained during step a) of solid / liquid extraction of the process as defined in any one of claims 1 to 5. [Claim 8] Method for producing a desolvated solid residue further comprising a step f) of desolvation of the solid residue recovered during step e) of the process as defined in claim 7 to recover on the one hand the solvent and on the other hand the desolvated solid residue .. [Claim 9] A method for producing a by-product comprising a step g) of transformation of the desolvated solid residue recovered during step f) of the process as defined in claim 8 to obtain the by-product. [Claim 10] The method of claim 9 wherein the by-product is selected from a flour, a protein concentrate, a protein isolate, a textured protein and mixtures thereof. [Claim 11] Crude oil obtained from a biological substrate, said crude oil comprising a polyphenol and 2-methyloxolane, and being characterized in that the mass concentration of polyphenol is greater than or equal to 100 ppm. [Claim 12] Crude oil according to claim 1 1 in which the mass concentration of 2-methyloxolane is from 0.5 ppm to 500 ppm. [Claim 13] Refined oil obtained from a biological substrate comprising a tocopherol, and being characterized in that the mass concentration of tocopherol is greater than or equal to 350 ppm. [Claim 14] Refined oil according to claim 13 further comprising 2-methyloxolane. [Claim 15] Solid residue from a biological substrate and comprising a polyphenol and a residual oil, characterized in that the mass concentration of polyphenol is less than or equal to 3000 ppm and the mass concentration of residual oil is less than or equal to 5%. [Claim 16] Solid residue according to claim 15 further comprising 2-methyloxolane. [Claim 17] Desolvated solid residue derived from a biological substrate comprising 2-methyloxolane, characterized in that the mass concentration of 2-methyloxolane is less than 1000 ppm. [Claim 18] By-product from a biological substrate comprising 2-methyloxolane, characterized in that the mass concentration of 2-methyloxolane is less than 1000 ppm. [Claim 19] By-product according to claim 18 chosen from a flour, a protein concentrate, protein isolate, a textured protein, and mixtures thereof.