METHOD FOR THE SELECTIVE SEPARATION OF AT LEAST ONE ORGANIC SUBSTANCE WITH AT LEAST ONE APOLAR RESIDUE AND THE USE OF THIS SUBSTANCE IN A FOOD, BEVERY, COSMETICS OR PHARMACEUTICAL PRODUCT

Description

[0001] The invention relates to a method for flavoring and / or stabilizing a product, in which a composition is added to the product to be flavored and / or stabilized, wherein the composition is produced by a method for the selective separation of an organic substance with at least one nonpolar residue.

[0002] The growing public awareness of a healthy lifestyle is boosting sales of non-alcoholic beverages, such as sparkling wines, soft drinks, and beers. This necessitates the search for new production techniques that allow beverages and food products to be produced not only with a palatable taste and aroma, but also entirely without the use of ethanol.

[0003] According to the European Food Information Regulation (EU No. 1169 / 2011), alcohol labeling is only mandatory for products containing 1.2% alcohol by volume or more. The term "alcohol-free" does not necessarily mean "without alcohol." For example, beer and wine advertised as "alcohol-free" may contain a maximum of 0.5% alcohol by volume. If a beverage other than wine or beer is advertised as "alcohol-free," it must contain absolutely no alcohol; the alcohol content must be 0.0% by volume. Otherwise, the "alcohol-free" label is considered misleading and deceptive to consumers. The term "alcohol" here refers to ethanol.

[0004] Important components of food, as well as of luxury goods, cosmetics, and pharmaceuticals, are obtained through extraction processes using solvents, often with the use of ethanol. However, the components obtained in this way can contain ethanol residues, meaning that products containing such components are not "alcohol-free."

[0005] Components obtainable by extraction in ethanol include not only volatile flavorings but also non-volatile flavorings such as bitter substances, oil, fat and / or wax fractions, dyes and adhesives, derived from plant and / or animal raw materials as well as other natural sources, for example flavorings obtained from microorganisms or fungi, and / or of synthetic origin.

[0006] These substances possess at least one nonpolar residue in their molecule, and can therefore be either nonpolar or amphiphilic. These substances are subsequently referred to collectively as "nonpolar organic compounds" and, for ease of reading, sometimes abbreviated as "AOS".

[0007] For many years there has been great interest in technologies that serve to recover, in particular, flavorings from dealcoholization and / or concentration processes of products such as beer / wine etc. or juices, or to remove unwanted substances such as bitter substances from citrus juices, but also to recover or remove nonpolar organic substances from cosmetics and pharmaceuticals, whereby the extracts obtained in this way can be used for alcohol-free re-aromatization of products or in other ways, for example as a coating agent in the case of waxes.

[0008] Four basic separation processes for obtaining desired and removing unwanted nonpolar organic substances, especially flavorings, dissolved in liquids such as water or ethanol are described in the literature: 1) Liquid-liquid extraction with organic solvents, in which the pure substances or the desired fraction are ultimately obtained by evaporating the solvent used. 2) Rectification of the nonpolar organic substance or flavoring-containing liquid. 3) Sorption processes in which nonpolar flavorings from the liquid phase are concentrated on a solid phase. 4) Precipitation and crystallization of solid flavoring components. 5) Direct extraction with supercritical gases, percolation. For fat, oil, and wax fractions from plant or animal source material, steam distillation and mechanical processes such as pressing are also applicable.

[0009] While extraction has a wide range of applications, the use of large quantities of organic, often chlorinated, solvents is highly questionable from an environmental and sustainability perspective. Furthermore, thermally labile substances can be irreversibly destroyed during solvent evaporation. Extracting flavor compounds from an ethanolic extract using liquid-liquid extraction is also difficult because ethanol itself is soluble in a variety of organic solvents. Direct re-aromatization with an ethanolic extract is not feasible, as the final product would not meet the label "alcohol-free" (i.e., with an alcohol content of 0.0% by volume).

[0010] During rectification, ethanol or water cannot be selectively removed, as they typically form azeotropes with the flavor compounds contained in the extract. Furthermore, thermally labile flavor-imparting components can be irreversibly destroyed.

[0011] In sorption processes, the desired enriched substances are eluted again using nonpolar media. Ethanol is frequently used as the eluent, especially in the food industry, which renders the extract unusable for 0.0 vol.% applications. Carbon dioxide is also used as an eluent. EP 3063260 describes a process for extracting two or more fractions from hop oil by treating the hop oil, loaded onto an adsorbing support, first with liquid carbon dioxide to separate the first fraction and then with supercritical carbon dioxide to separate the second fraction. Further fractions can be separated by combining the supercritical carbon dioxide with a co-solvent. While elution with liquid / supercritical carbon dioxide avoids the aforementioned undesirable use of alcohols, significant yield losses occur during gas discharge.The construction and operation of a CO2 extraction plant are expensive and have specific safety requirements.

[0012] Mechanical processes such as pressing are primarily used in oil extraction and do not require solvents.

[0013] However, these methods are not universally applicable for obtaining all non-polar organic substances, especially all oils and waxes from plants.

[0014] The object of the invention is therefore to provide a process for flavoring and / or stabilizing a product based on the selective extraction of at least one nonpolar organic substance, in particular at least one flavoring agent, preferably from a natural source such as plant or animal products, enabling the use of the extracted substances in foods, beverages, cosmetics, and / or pharmaceutical products that can be labelled "alcohol-free." In particular, such products should meet the quality requirements for the designation "FTNS" ("From the named source"). The aim is to enable so-called "clean label" products. A further object of the invention is to enable the selective extraction of non-volatile flavoring agents, such as bitter substances, as well as the selective extraction of oil, fat, and / or wax fractions from plant and / or animal starting materials.

[0015] Several processes are known in the art in which cyclodextrins are used to remove unwanted substances from food systems. Cyclodextrins are water-soluble and toxicologically and environmentally safe. For example, cyclodextrins are used to remove cholesterol from butter (AU 638531B2) or from eggs (EP326469A, EP475451B1). Such removal processes typically consist of two stages: first, the cyclodextrin is mixed with the food system, forming a complex between the cyclodextrin and the unwanted substance (host complex); and then this complex is removed from the food system. The cyclodextrin can be released from the complex (decomplexation) by, for example, treatment with hot water or alcohol (40–100 °C) and then reused in the removal process.

[0016] German patent application DE 40 29 287 A1 refers to the aforementioned European patent 3 26 469 and relates to a process for producing cholesterol-reduced egg yolk. After separating the cholesterol- or cholesterol ester-loaded β-cyclodextrin from the diluted egg yolk, the added water is removed from the egg yolk, and the remaining amounts of β-cyclodextrin present in the egg yolk are enzymatically degraded using α-amylases and / or CT gases.

[0017] In the publication CHARMILA C RATNASOORIYA ET AL: "Extraction of phenolic compounds from grapes and their pomace using cyclodextrin", FOOD CHEMISTRY, ELSEVIER LTD, NL, Vol. 134, No. 2, February 2, 2012, pages 625-631, the extraction of phenolic components from grapes and their pomace using β-cyclodextrin is described, and it is explained that cyclodextrins have the ability to form host-guest complexes to encapsulate a wide range of components. Accordingly, β-cyclodextrin is used in the food industry as an encapsulating agent to protect desired components or to mask unwanted components. The publication RAJHA HIBA N. ET AL: "[beta]-Cyclodextrin-Assisted Extraction of Polyphenols from Vine Shoot Cultivars", JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, Vol. 63, No. 13, April 8, 2015, pages 3387-3393 also concerns the β-cyclodextrin-assisted extraction of polyphenols from vine shoots.In both studies, HPLC analyses with acetonitrile were performed to determine the phenol concentration.

[0018] The two publications mentioned above are referenced in the paper EL DARRA NADA ET AL: "Comparative Study between Ethanolic and [beta]-Cyclodextrin Assisted Extraction of Polyphenols from Peach Pomace", INTERNATIONAL JOURNAL OF FOOD SCIENCE, Vol. 2018, January 1, 2018, pages 1-9. This publication describes a comparative study between ethanolic and β-cyclodextrin-assisted extraction of polyphenols from peach pomace.

[0019] European patent application EP 0 110 170 A1 relates to a honey powder that can retain its natural flavorings for an extended period and to a method for its production using cyclodextrins. According to the application, honey can be mixed with a hot saturated solution of cyclodextrin in water, the resulting homogeneous solution frozen and lyophilized, followed by pulverization. The honey powder releases its flavorings only when the baked goods made from it are consumed.

[0020] Document DE 693 27 335 T2 concerns a method for removing residual cyclodextrin. For example, β-cyclodextrin is added to coffee, and after treatment at 60°C for one hour, the solution is cooled to room temperature, and a complex of caffeine and cyclodextrin is filtered out. The caffeine-free coffee solution of the filtrate is then treated with various enzymes. According to this document, the aim is to create a method for completely removing all cyclodextrin residues from a food system.

[0021] The use of cyclodextrins in the currently known extraction of flavor compounds has the disadvantage that the solvent used for decomplexation must be used in larger volumes and at a higher temperature, which leads to a dilution of the flavor concentration and also results in additional technical effort and high costs. If ethanol is to be used as the solvent for decomplexation, in addition to dilution of the flavor concentration, a further problem will arise, namely that this ethanol cannot be removed without affecting the flavor profile.

[0022] The inventors have surprisingly discovered that cyclodextrins in aqueous and alcoholic media can reversibly absorb components obtainable by extraction in ethanol, particularly for foodstuffs, such as flavorings, bitter substances, oils, fats and waxes, adhesives and dyes, into their cavity.

[0023] Such components possess at least one nonpolar residue and can therefore be either nonpolar or amphiphilic. An amphiphilic component possesses at least one polar part in addition to at least one nonpolar part. Organic substances with at least one nonpolar residue—even if they also possess a polar part and are particularly amphiphilic—are referred to below as "nonpolar organic substances." This term is sometimes abbreviated as "AOS." The organic substances with at least one nonpolar residue obtained by extraction in ethanol, as described in the invention, are more hydrophobic in water than ethanol.

[0024] The invention thus made it possible to selectively convert nonpolar organic substances from a starting mixture into a complex of cyclodextrin and the nonpolar organic substance by selecting a suitable cyclodextrin.

[0025] The invention thus solves the aforementioned problems in a very simple way using a method according to claim 1.

[0026] The invention presents a method for flavoring and / or stabilizing a product that is a food, beverage, cosmetic or pharmaceutical product, in which a composition is added to the product to be flavored and / or stabilized, wherein the composition is produced by a method for the selective separation of at least one organic substance with at least one nonpolar residue (AOS) and / or at least one or more flavoring substances comprising: (a) Providing a starting mixture containing at least one organic substance with at least one nonpolar residue (AOS) and / or at least one flavoring substance or several flavoring substances and optionally at least one solvent, (b) Contacting the starting mixture with at least one cyclodextrin, wherein the organic substance with at least one nonpolar residue is selected from the group comprising volatile flavoring substances, non-volatile flavoring substances such as bitter substances, oil, fat and / or wax fractions, colorants and adhesives, in particular from vegetable and / or animal starting materials, as well as mixtures of at least two of the aforementioned substances, and wherein at least one solvent, in particular water, is added in step a) and / or in step b) and / or after step b),and wherein the contact of the at least one cyclodextrin with the at least one organic substance having at least one nonpolar residue of the starting mixture leads to the formation of at least one cyclodextrin-AOS complex and / or at least one cyclodextrin-flavoring complex in a liquid, in particular aqueous, phase, comprising the further steps of (c) separation of the cyclodextrin-AOS complex and / or the cyclodextrin-flavoring complex from the liquid phase and (d) enzymatic treatment of the separated cyclodextrin-AOS complex and / or the cyclodextrin-flavoring complex, wherein a composition loaded with at least one organic substance having at least one nonpolar residue (AOS) and / or with at least one flavoring is obtained, , wherein the enzymatic treatment is carried out with at least one enzyme selected from the group comprising enzymes with amylase activity, preferably alpha-amylase, particularly preferably with fungal-alpha-amylase, pulluanase and / or isoamylase, as well as mixtures of at least two of the aforementioned enzymes.

[0027] The terms "the cyclodextrin" or "a cyclodextrin" refer to the total number of cyclodextrin molecules in the respective system under consideration, such as the starting mixture or the liquid phase. This applies accordingly to the expression "the cyclodextrin-AOS complex" or "a cyclodextrin-AOS complex".

[0028] The process for the selective separation of at least one organic substance with at least one nonpolar residue (AOS) and / or at least one or more flavorings yields a composition loaded with at least one organic substance with at least one nonpolar residue, such as a flavoring or bittering agent. This enables reversible protection of, for example, thermally labile substances, which can be removed as soon as it is no longer needed.

[0029] Depending on the material system in which the at least one nonpolar organic substance is present in the starting mixture, no addition of solvents or further solvents is necessary to carry out the process according to the invention. The starting mixture can, for example, already be a solution itself, or, after the addition of cyclodextrins dissolved in water in step b), contain sufficient solvent from this water.

[0030] The quality of complex formation can be influenced by adjusting the solvent content, particularly the water and / or ethanol content, of the initial mixture. Therefore, the invention offers the possibility of adding at least one solvent in step a) and / or in step b) and / or after step b), depending on the application. A person skilled in the art will adjust the solvent content to optimize the quality of complex formation for the specific application. The choice of cyclodextrin, the temperature, and / or the energy input during complexation, for example by adjusting the stirrer speed, also offer possibilities for influencing the quality of complex formation.

[0031] Within the scope of the invention, the conditions for complex formation can be tailored to the nonpolar organic substances to be separated. In particular, the extraction conditions can be adapted within the scope of the invention such that, for example, bitter substances can be selectively complexed and isolated, or remain unaffected.

[0032] In an advantageous embodiment of the invention, it is provided that in step a) and / or in step b) and / or after step b), in particular by adding water, a water content in the range of 15 vol.% to 35 vol.%, preferably from 20 vol.% to 30 vol.%, and / or, in particular by adding ethanol, an ethanol content of at least 40 vol.%, preferably an ethanol content in the range of 40 vol.% to 60 vol.%, is adjusted.

[0033] Ethanol-containing extracts of nonpolar organic substances, particularly flavorings and / or bitter substances, are ideally suited for use in flavorings for addition to foods, beverages, cosmetics, or pharmaceutical products, as they possess a natural, complex flavor profile very similar to the starting material. The invention enables the preservation of this complexity after dealcoholization for an alcohol-free product by adjusting the water and / or ethanol content during the complexation of the nonpolar organic substance using cyclodextrin.

[0034] Process optimizations have shown that dilution with water to 15 to 35 vol%, preferably in the range of 20 to 30 vol%, significantly increases the complexation efficiency of cyclodextrins for volatile aroma components. However, depending on the specific compound, the addition of water can surprisingly result in non-volatile aroma components, such as bitter substances, no longer being complexed. This alters the natural aroma profile of the extract. Within the scope of the invention, it was surprisingly discovered that complexation of bitter substances can nevertheless be achieved with an ethanol content of at least 40 vol% in the liquid phase during complex formation with cyclodextrin.

[0035] The invention thus enables the preservation of the characteristic aroma profile from an ethanolic extract, which contains not only volatile but also non-volatile aroma compounds such as bitter substances. Furthermore, the invention offers the possibility of selectively removing non-volatile aroma compounds, such as bitter substances, from an extract containing nonpolar organic substances by means of conditions unfavorable to their complexation, in particular an ethanol content below 40% by volume.

[0036] Although the use of cyclodextrins (CDs) in food systems is known, to the inventors' knowledge they have never before been used for the selective extraction of flavorings or other organic substances with at least one nonpolar residue. The invention thus provides a process in which cyclodextrins are used to obtain alcohol-free (i.e., ethanol-free), declaration-free FTNS (FTNS = "from the named source") flavorings or bitterings from aqueous, but also (and especially) from ethanolic extracts.

[0037] The invention thus provides a process that enables the separation / obtaining of organic substances with at least one nonpolar residue and / or at least one or more flavorings from an aqueous, ethanolic, or other extract in a concentrated form using cyclodextrins, while reducing the technical effort (or solvent consumption) and allowing the entire process to be carried out at mild temperatures. These obtained nonpolar organic substances, such as flavorings, are suitable for use in foods, beverages, cosmetics, and / or pharmaceutical products that can be labelled "alcohol-free."

[0038] The invention provides a process that enables the selective recovery of oil, fat, and / or wax fractions, particularly from plant and / or animal starting materials. Cyclodextrin has proven to be a surprisingly simple and suitable auxiliary agent for this purpose, as it is both water-soluble and capable of binding the target substances. The target substance can then be removed from the cyclodextrin-AOS complex without the need for any further solvents.

[0039] The invention allows all organic substances with at least one nonpolar residue to be obtained by extraction from the starting mixture using cyclodextrins that are hydrophobic enough to form a complex with alpha-, beta-, gamma-, and / or delta-cyclodextrin as the host. In other words, the organic substance in question, with at least one nonpolar residue, is hydrophobic enough to transfer from a solution into the cavity of alpha-, beta-, gamma-, and / or delta-cyclodextrin. According to the invention, it is not always necessary for the entire nonpolar substance to be complexed into the cavity of the cyclodextrin. For example, with fats and organic acids, essentially only the hydrophobic part of the substance is complexed into the cavity of the cyclodextrin. The polar part of the substance protrudes from the cavity, according to a model.Even with amphiphilic substances, the hydrophobic part is absorbed into the cavity, while the hydrophilic part remains virtually untouched.

[0040] The process according to the invention does not subject the nonpolar organic substances to high thermal stress. Within the scope of the invention, it is therefore provided that the process is carried out at maximum temperatures in the range of at least 40°C to a maximum of 70°C, preferably at a temperature of at least 40°C to a maximum of 55°C. These temperatures relate in particular to step d), which is explained below. In step b), the temperatures are preferably lower, in particular in the range of 4°C to 10°C, preferably at 6°C.

[0041] The invention offers the advantage that it can be used for different fields of application, since at least one organic substance with at least one nonpolar residue can be selected from the group which contains secondary metabolites, This includes phytochemicals, in particular secondary plant metabolites, volatile flavorings, non-volatile flavorings such as bitter substances, colorants, oil, fat and / or wax fractions, adhesives, especially of natural origin, preferably from plant and / or animal starting materials, as well as mixtures of at least two of the aforementioned substances. Natural flavorings, including those obtained from microorganisms or fungi, are of particular interest. Phenolic substances can be important for stabilizing colorants.

[0042] Within the scope of the invention, the method comprises the further step: (c) separation of the cyclodextrin-AOS complex and / or the cyclodextrin-flavoring complex from the liquid phase, in particular from the solvent.

[0043] This allows the (still complexed) nonpolar organic substance, in particular the flavoring agent, to be concentrated. Separation of the cyclodextrin-AOS complex and / or the cyclodextrin-flavoring agent complex from the liquid phase results in a separated solid phase containing at least the cyclodextrin-AOS complex and / or the cyclodextrin-flavoring agent complex. The liquid phase contains water and solvent, in particular ethanol, with which the nonpolar organic substance, such as at least one flavoring agent, was provided in the starting mixture and / or added in the process according to the invention.

[0044] After step c), a substantially solvent-free solid phase remains. "Substantially solvent-free" means, in particular, that solvents such as water and / or ethanol are adsorbed onto the solid phase but are no longer freely present within it. Depending on the application, the removal of water in step c) can also separate unwanted components from the initial mixture from the filter cake, similar to the function of a washing step.

[0045] The separation of, for example, water, but also other liquids, from the solid phase, particularly from the filter cake, can be aided by drying according to the invention. In a simple embodiment, the separated solid phase, for example as a filter cake, can be subjected to a flow of compressed air, particularly at a pressure of 2 bar, towards the filter layer. An inert gas can also be used instead of compressed air.

[0046] Known methods for removing alcohol by means of evaporation or reverse osmosis result in the removal of other important, particularly volatile, flavor compounds along with the ethanol, which are crucial for the characteristic taste of the extract. Furthermore, thermally labile substances can be destroyed, or undesirable changes in taste can occur. The method described in the invention allows for the highly selective removal of ethanol from the extract. The complexity of the extract is preserved by ensuring that both volatile and non-volatile flavor components are complexed equally.

[0047] Within the scope of the invention, the process comprises the further step (d) Enzymatic treatment of the separated cyclodextrin-AOS complex and / or the cyclodextrin-flavoring complex, wherein a composition loaded with at least one organic substance with at least one nonpolar residue (AOS), in particular with at least one flavoring substance, is obtained.

[0048] Surprisingly, it has been shown that the enzymatic treatment not only decomposes free cyclodextrins, but also cyclodextrins that are bound in a complex with at least one nonpolar organic substance such as a flavoring agent, thereby enriching the nonpolar organic substance, in this example the flavoring agent (without impairment), in the resulting composition.

[0049] In addition to treatment with the enzymes described in more detail below, this degradation of the cyclodextrin and thus the release of the nonpolar organic substance from the complex can also be achieved within the scope of the invention by the use of organisms from the group which includes such yeasts, fungi and mixtures thereof that are able to degrade cyclodextrin.

[0050] The treatment of the cyclodextrin-AOS complex and / or the cyclodextrin-aromatic complex to obtain the nonpolar organic substance can be supported by a further step d1) diluting the cyclodextrin-AOS complex and / or the cyclodextrin-aromatic complex separated in step c) with water before the particularly enzymatic treatment in step d).

[0051] Due to the preceding step c) described above for the separation of water, the water added in step d1) for enzymatic treatment is then free of unwanted components.

[0052] In an advantageous embodiment of the invention, the pH value for the enzymatic reaction can also be adjusted to suit the enzymes used, for example, to a value in the acidic range, particularly pH 4.5. Within the scope of the invention, the activity of the enzyme can be influenced by adjusting the pH value. This effect can be used to degrade one type of cyclodextrin by adjusting the pH value and then, by changing the pH value, another type of cyclodextrin with the same enzyme.

[0053] The following describes the advantages of the process according to the invention using the example of flavoring substances as a nonpolar organic compound. These advantages also apply to the other nonpolar organic compounds mentioned above: The process according to the invention makes it possible to obtain a composition from a diluted starting mixture in which flavoring substances are present in a concentrated form. The separation can be gentle and complete. Furthermore, thanks to this process, it is possible to obtain flavor-laden compositions from starting mixtures even with a very high alcohol content (namely up to 80% by volume), which can be used in foods, beverages, cosmetic products, and / or pharmaceutical products that can be labelled "alcohol-free."

[0054] Because the process according to the invention can be carried out at a temperature not exceeding 55 °C or even room temperature, flavorings can be obtained that are highly and moderately volatile (i.e., with an evaporation number < 10 or from 35 to 10, respectively) and / or thermally labile. A further advantage of the process according to the invention is that the α-, β-, and γ-cyclodextrins used can be partially and selectively hydrolyzed with different amylases, allowing for a cascading release of flavorings. These points will be discussed in more detail below.

[0055] In a further development of the invention, step d) is carried out such that the cyclodextrin concentration in the composition loaded with at least one nonpolar organic substance (AOS), in particular with at least one flavoring substance, is less than 0.5 wt.%, preferably less than 0.1 wt.%. Such a low cyclodextrin content has no effect on the use of the obtained nonpolar organic substance (AOS).

[0056] The cyclodextrin concentration can be adjusted relative to the total mass of the composition that constitutes the final product in the selected embodiment of the process according to the invention. This composition can, for example, after steps a), b), c), and d), be a mixture of water, flavoring, cyclodextrin degradation products, enzyme, and residual cyclodextrin with a proportion of less than 0.1 wt.% from an aqueous flavoring solution. If components such as water and enzymes are removed in further process steps, the enzymatic treatment can be carried out in such a way that the cyclodextrin concentration is further reduced and therefore lies below 0.1 wt.% in the water- and enzyme-free product.

[0057] Adjusting the cyclodextrin concentration can be aided by removing undesired components from the final product, which may include undegraded cyclodextrin-AOS complexes. According to a further advantageous embodiment of the invention, the process comprises a further step (e) filtering the mixture resulting from the particularly enzymatic treatment of the cyclodextrin-AOS complex, yielding a composition loaded with at least one organic substance with at least one nonpolar residue (AOS). Step (a)

[0058] In the present invention, "flavoring agent" refers to an organic substance, particularly volatile at room temperature, that evokes or modifies odor and taste perceptions. Flavoring agents are frequently alcohols, acids, esters, lactones, aldehydes, ketones, acetals, ketals, ethers, epoxides, and their analogous sulfur compounds. Other examples include oxygen, nitrogen, and sulfur heterocycles, heteroaromatics (e.g., alkylpyrazines), amines, and amides. Simple or complex, saturated and unsaturated, aliphatic and cycloaliphatic compounds, aromatics, and terpenes, etc., are also considered flavoring agents. The most important types of flavoring agents are natural, nature-identical, and artificial. Within the scope of the present invention, so-called natural flavoring agents, where the starting material is of plant or animal origin, are preferred. Examples of animal-derived starting materials include honey, milk, meat, bones, and body fluids.The starting material of plant origin consists of plants or plant parts, such as flowers, buds, leaves, stems, stalks, bark, roots, tubers, bulbs, rhizomes, fruits, nuts, berries, and seeds, as well as fruits and vegetables. These starting materials can be present, for example, in fresh, cooked, sprouted, dried, fermented form, or in a form prepared for consumption as food or beverages (e.g., beer, wine, sparkling wine, spirits such as whiskey, etc.). These starting materials can either be used directly as a starting mixture in the process according to the invention, or they can be processed before being used as a starting mixture in at least one process known to those skilled in the art. These processes include, for example, dissolving, dispersing, purification, mashing, soaking, fermentation, and / or separation processes such as extraction, filtration, etc.Examples of plants that provide ethanolic extracts containing bitter substances are gentian, chiretta, and wormwood.

[0059] The starting mixture may contain, in addition to such "flavoring substances," other components of the starting material of plant or animal origin. In particular, the starting mixture may contain one or more nonpolar organic substances, including non-volatile flavoring substances such as bitter substances, oil, fat and / or wax fractions, especially from plant and / or animal starting materials, as well as mixtures of at least two of the aforementioned substances.

[0060] The starting mixture can be in the form of a solid or liquid dispersion, in particular a powder, a solution, a suspension, or an emulsion, according to the invention. In principle, the starting mixture can contain any solvent in which the nonpolar organic substances to be separated dissolve and which can be readily absorbed into or displaced from the cavity of the cyclodextrin molecule containing at least one substance to be separated. In step a) and / or in step b) and / or after step b), water, C1-C4 alcohols, diethyl ether, acetone, etc., or mixtures thereof, can be used as solvents according to the invention. Preferably, a solvent selected from the group consisting of water, ethanol, or mixtures thereof is used. A solvent can also be added in step a) during the preparation of the starting mixture.For example, depending on which substances are to be separated, starting mixtures containing ethanol and / or water can also be extracted directly using cyclodextrin according to the invention.

[0061] In one embodiment of the invention, at least one of the above-mentioned solvents can be added in step (b) in addition to the addition in step (a).

[0062] The preferred solvent content is less than 100% by weight. The person skilled in the art will select the solvent content and the mixing ratio of two or more solvents according to the solubility of the flavoring substance(s) to be separated, the effectiveness of complex formation, and taking process efficiency into account. A starting mixture in which the at least one nonpolar organic substance, in particular the flavoring substance(s), is dissolved in a minimal amount of solvent is particularly preferred.

[0063] The higher the solvent concentration, the greater the solubility of the flavoring to be separated and of the cyclodextrin used, thus increasing the rate of complex formation. However, excessively dilute solutions should be avoided, as the process becomes uneconomical and the probability of contact and interaction between flavoring molecules and cyclodextrin becomes too low. According to one model, these relationships are based on the fact that at higher concentrations of nonpolar organic substances, complexes of these substances with cyclodextrin are preferentially formed due to the driving gradient. If the dilution is too great, the driving force for complex formation is no longer sufficient.

[0064] Preferably, a starting material in a form prepared for consumption as food or beverages (e.g., beer, wine, sparkling wine, spirits, etc.) is used, which is subject to extraction, preferably solid-phase extraction (English: solid-phase extraction). solid phase extraction The solid-phase extract is subjected to solvent extraction (SPE, older term also 'sorbent extraction'), whereby the resulting solvent-containing solid-phase extract is used as the starting mixture. An ethanol-containing solid-phase extract is particularly preferred, in which the ethanol content can be up to 80% by volume based on the total volume of the solid-phase extract. The flavoring substance(s) content is at least approximately 0.1% by weight, preferably from 0.5% to 8% by weight, based on the total weight of the solid-phase extract.

[0065] Seeds of any kind, especially cereals, as well as products obtained from cereals, such as malt, wort, mash, beer, or the like, constitute another preferred starting material. The cereals can be selected, for example, from the group comprising barley, wheat, rye, spelt, maize, oats, rice, millet, triticale, and mixtures thereof. This starting material can either be used directly as a starting mixture in the process according to the invention or processed in at least one process known to those skilled in the art (so). Particularly preferred as a starting material is wort and / or mash subjected to fermentation, with the resulting fermentate being used as the starting mixture. The content of flavoring substance(s) is at least approximately 0.01 wt.%, preferably from 0.1 to 8 wt.%, based on the total weight of the fermentate. Step (b)

[0066] In step (b) of the process according to the invention, the cyclodextrin-AOS complexes are produced by bringing at least one cyclodextrin into contact with the starting mixture. The components can be mixed either in a powdered state or in a suspension and / or in an emulsion and / or in a solution.

[0067] Several methods for the preparation of guest-host complexes are known, with the most common being preparation in solvents (coprecipitation and complexation in a suspension "slurry method") or kneading methods (see e.g. SK et al. / / Research Journal of Pharmaceutical, Biological and Chemical Sciences., 2013, Vol. 4, No. 2, pp. 1694-1720).

[0068] The invention uses coprecipitation or complexation in a suspension ("slurry method") in which at least one cyclodextrin is added to an aqueous or ethanol-containing solution or suspension of the starting mixture and precipitating complexes are separated.

[0069] The term "cyclodextrin" can be a substituted or unsubstituted α-cyclodextrin, at least a substituted or unsubstituted β-cyclodextrin, at least a substituted or unsubstituted γ-cyclodextrin, or at least a substituted or unsubstituted δ-cyclodextrin, or mixtures thereof, preferably a substituted or unsubstituted γ-cyclodextrin. Since the use of substituted cyclodextrins as degradation products of the optional process step d) of enzymatic treatment can generate substances that are undesirable in food and can be removed in a further separation step, the use of unsubstituted cyclodextrins is preferred in such cases.

[0070] According to one model, gamma-cyclodextrin, compared to the other cyclodextrins mentioned, exhibits a good balance between solubility in water and / or ethanol on the one hand and its cavity size on the other. Furthermore, particularly good enzymatic degradation has been observed, which the model attributes to the flexibility of the gamma-cyclodextrin backbone due to its size. Following the lock-and-key principle, this molecule can thus fit particularly well into the active pocket of the enzyme.

[0071] A mixture of 10 wt% substituted or unsubstituted β-cyclodextrin and 90 wt% substituted or unsubstituted γ-cyclodextrin is particularly preferred, especially with regard to the separation of the cyclodextrin-AOS complex from the liquid phase resulting from the starting mixture. This will be discussed in more detail below. The use of a high proportion, up to and including exclusively γ-cyclodextrin, is preferred when non-volatile, nonpolar organic substances, and in particular bitter substances, are to be obtained by the extraction according to the invention using cyclodextrins. The invention thus also offers the possibility of selectively removing non-volatile flavoring substances, such as bitter substances, from an extract containing organic substances with at least one nonpolar residue under conditions unfavorable for their complexation, in particular an ethanol content below 40 vol% and / or the exclusive use of gamma-cyclodextrin.

[0072] According to the invention, a mixture of different cyclodextrins, such as beta-cyclodextrin with gamma-cyclodextrin, makes it possible to complex bitter substances predominantly, or almost exclusively, in the gamma-cyclodextrin molecules, and other flavors predominantly, or almost exclusively, in the other cyclodextrin molecules. Figuratively speaking, the "guests" seek out the most suitable "hosts" for complex formation. In this way, the composition of the removed organic substances with at least one nonpolar residue can be influenced by the ratio of the different cyclodextrins.

[0073] Cyclodextrin can be used in the form of a mixture with water, but also in at least one of several other solvents such as glycerin, propylene glycol, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), and ethylene glycol. Cyclodextrin is preferably used in a solid form, so that it can be easily handled and added to the starting mixture as a solid. Furthermore, using cyclodextrin as a solid avoids additional dilution of the starting mixture by solvent introduced with the cyclodextrin.

[0074] The amounts of cyclodextrin are varied within further limits, but preferably between approximately 1 and approximately 100 wt.%, preferably between approximately 3 and approximately 50 wt.%, and particularly preferably between approximately 3 and approximately 20 wt.%, based on the amount of the starting mixture, are used.

[0075] The mixing of at least one cyclodextrin with the starting mixture is carried out using any device known to those skilled in the art, such as a mechanical stirrer, mechanical disperser, or ultrasonic disintegrator. Those skilled in the art will select the temperature and holding time parameters to optimize the effectiveness of complex formation and to ensure process efficiency.

[0076] In general, temperature and holding time can be varied widely. The preferred temperature range is between approximately 4 °C and 25 °C, preferably between approximately 4 °C and 15 °C, particularly preferably between approximately 4 °C and 10 °C, and most preferably at a temperature of approximately 6 °C. The holding time can be up to 5 days, preferably over a period of 20 minutes to 72 hours, and particularly preferably over a period of up to 48 hours.

[0077] Within the scope of the invention, it was found that cyclodextrins in aqueous and alcoholic media can reversibly incorporate organic substances with at least one nonpolar residue into their cavity, whereby approximately 60 to 99 wt% of the flavorings present in the starting mixture can be selectively complexed, while the remaining components of the starting mixture largely remain in the liquid phase. This finding of the reversible incorporation of organic substances with at least one nonpolar residue is utilized by the process according to the invention for the efficient selective separation of these substances. Step (c)

[0078] In step (c), the cyclodextrin-AOS complexes formed in step (b) are separated from the liquid phase. The cyclodextrin-AOS complexes formed are quite stable, meaning that the nonpolar organic substance bound within the complex, such as a flavoring agent, has a low tendency to escape the complex or the cyclodextrin cavity. The complexes are therefore stable enough to be concentrated in the aqueous medium using filtration techniques. In principle, they can be separated using any conventional solid-liquid separation process without releasing the gas. Preferred separation methods for the cyclodextrin-AOS complex in step (c) are filtration and / or sedimentation and / or centrifugation, with filtration, such as vacuum or nanofiltration or ultrafiltration, being particularly preferred.Reverse osmosis is also a possible separation method for the separation of the cyclodextrin-AOS complex in step (c).

[0079] In a further advantageous embodiment, the method according to the invention comprises a further step c1) allowing the mixture to rest before separating the cyclodextrin-AOS complex from the liquid phase, wherein the resting phase according to step c1) is carried out in particular for a duration of up to 24 hours, preferably for a duration of up to 12 hours, and particularly preferably for a duration of 2 hours.

[0080] Allowing the starting mixture containing cyclodextrin to settle, in which the cyclodextrin-AOS complex forms or has already formed, facilitates the separation of the complexes. Surprisingly, it has been found that this settling process results in faster sedimentation and improved filtration properties. The settling process induces sedimentation, and this sedimentation allows a clear phase, essentially free of cyclodextrin-AOS complexes, to be collected above the sediment containing the cyclodextrin-AOS complex. This reduces the total volume that needs to be separated, for example, by filtration, in step c). Step c) can also be performed solely by collecting the supernatant to obtain a phase containing the cyclodextrin-AOS complexes.

[0081] Filter aids are commonly used to improve filtration in solid-liquid separation processes. These can be incorporated into filter plates, filter sheets, or filter cartridges, or added directly to the suspension to be filtered as a sedimentation aid (diatomaceous earth, cellulose, bentonite, or other cationic flocculants). Physically, sedimentation and thus separation can be accelerated, for example, by using centrifuges. Other processes using filter aids include precoat filtration. The most important examples are diatomaceous earth filtration and perlite filtration. In this process, a filter sheet is precoated onto a horizontal or vertical support layer. The actual filtration is carried out with the continuous addition of the filter aid to prevent clogging of the filter.

[0082] The filter aids ensure the filter's permeability and thus prevent, especially in filters consisting of a single filtration layer, the filter surface from gradually becoming blocked by the accumulating filter cake. The filtrate is usually the desired product, and the retentate or filter cake is discarded.

[0083] The invention aims to eliminate the need for the aforementioned common filter aids, as the cyclodextrin-AOS complex is separated as a filter cake, which in this case represents the desired "product" of separation step c). If the aforementioned common filter aids were used, they would combine with the cyclodextrin complex to form the retentate. The filter aid would then have to be separated from the filter cake in a further process step. This is possible within the scope of the invention. A significant improvement in economic, ecological, and nutritional terms is achieved by avoiding the mixing of the final product with filter aids.

[0084] Studies on the sedimentation behavior and filtration properties of different cyclodextrins have shown that complexes with β-cyclodextrin sediment faster than analogous complexes with γ-cyclodextrin. Furthermore, complexes with β-cyclodextrin exhibit better filtration behavior than analogous complexes with γ-cyclodextrin, as they agglomerate to such an extent that the filter surface is significantly less blocked, or almost not blocked at all.

[0085] Products obtained with β-cyclodextrin do not always have positive sensory properties. Flavorings obtained with γ-cyclodextrin, on the other hand, are of higher sensory quality, but they sediment slowly and clog the filter surface very quickly.

[0086] Therefore, mixtures of beta-cyclodextrin with other cyclodextrins were investigated, and surprisingly, it was found that even the use of relatively small amounts of beta-cyclodextrin, for example, 10 wt% beta-cyclodextrin relative to the total mass of cyclodextrins used, can significantly accelerate sedimentation. The invention provides a means of accelerating sedimentation or centrifugation by adding beta-cyclodextrin to alpha-cyclodextrin and / or gamma-cyclodextrin and / or delta-cyclodextrin, preferably to gamma-cyclodextrin.

[0087] In an advantageous embodiment, the proportion of beta-cyclodextrin in the total mass of cyclodextrins is in the range of 0.5 wt.% to 60 wt.%, preferably in the range of 2 wt.% to 50 wt.%, and particularly preferably in the range of 5 wt.% to 15 wt.%. Thus, within the scope of the invention, the composition of the cyclodextrins can be selected for the respective application with regard to specific organic substances or substances with at least one nonpolar residue to be obtained from a starting mixture and / or with regard to the rate of the separation step.

[0088] In combination with 90 wt% gamma-cyclodextrin, sedimentation was accelerated, for example, from one day to two hours. A flavoring obtained using a mixture of 10 wt% beta-cyclodextrin and 90 wt% gamma-cyclodextrin had the same sensory quality as the corresponding flavoring extracted from pure gamma-cyclodextrin.

[0089] The sedimentation rate was measured using an ethanolic beer extract containing 60% ethanol by volume and an aroma concentration in the range of 5 to 8% by weight. In this case, the term "aroma" encompassed all aroma-active components detected in the GC spectrum. The suspension obtained after complexation, containing cyclodextrin-aroma complexes in the ethanolic extract, was simultaneously transferred to identical graduated cylinders. The sediment volumes were read from the graduated cylinder scale at 30-minute intervals. This method allows the sedimentation behavior of substances complexed in cyclodextrin according to the invention to be determined.

[0090] Separating the cyclodextrin-AOS complex from the liquid phase resulting from the starting mixture can be an important step in increasing the yield of organic matter containing at least one nonpolar residue. For example, if a flavoring agent is to be obtained from an ethanol-containing starting mixture, solid-liquid separation of the cyclodextrin-flavoring agent complexes from the ethanolic medium can influence the efficiency of the subsequent, particularly enzymatic, treatment of the complexes.

[0091] Solid-liquid separation produces a filter cake containing the cyclodextrin-AOS complex. If this filter cake is not sufficiently dry, the remaining alcohol can hinder the subsequent enzymatic degradation of the cyclodextrin complex when separating flavorings from an ethanolic medium. Furthermore, in single-layer filtration, the filter cake gradually builds up, potentially blocking the filter layer and rendering the separation step uneconomical due to time constraints. Filter aids can prevent the accumulating filter cake from blocking the filter layer in such situations.

[0092] It was surprising for the inventors to discover that β-cyclodextrin can serve not only as an extraction solvent for a nonpolar organic substance complexed as a guest, but also as a filter aid. Advantageously, β-cyclodextrin as a filter aid can remain in the filter cake and be enzymatically degraded in a further step into substances safe for use in food. Thus, β-cyclodextrin as a filter aid is largely similar to the substance used for complexing nonpolar organic substances and has the same chemical and physical properties.

[0093] The inventors were surprised to discover that beta-cyclodextrin, like gamma-cyclodextrin, binds flavorings and also acts as a filter aid. In the process according to the invention, beta-cyclodextrin is used both as an extraction and filter aid. It can then be broken down using enzymes, simultaneously releasing the complexed flavorings. The only breakdown products of the cyclodextrins are mono-, di-, and oligosaccharides, which are commonly found in foods and beverages and can therefore remain in the product. This will be discussed in more detail below. Thus, the invention prevents the problem described above of filter aids mixing with and remaining in the product.

[0094] The product thus exhibits the same nutritional purity and quality as when using gamma-cyclodextrin alone, while simultaneously offering significantly improved processability. Here, quality refers specifically to the sensory quality of the nonpolar organic substance. For example, in flavorings, an excessively high beta-cyclodextrin content leads to a change in the flavor profile, as some flavor components are preferentially complexed at the expense of others. However, the sweetness from the cyclodextrin breakdown products is usually negligible when considering the final dosage.

[0095] This invention was tested on the ethanolic beer extract described above. The test comprised the complexation of flavorings in gamma / beta-cyclodextrin mixtures, measurement of the sedimentation rate, drying of the filter cake, enzymatic degradation of cyclodextrins to release the flavorings, analytical determination of the flavor concentration and flavor profile, and sensory evaluation of the samples thus produced. The flavor concentration was quantified as the sum of all flavor components typical of beer using gas chromatography. The amount of residual cyclodextrin was measured using HPLC with an RI detector. As little as 10 wt% beta-cyclodextrin in the cyclodextrin mixture used is sufficient to accelerate the sedimentation rate from one day to two hours. The final product is an aqueous flavor extract of flavorings typical of beer that is more concentrated compared to the initial mixture.This was successfully added to a beer with an alcohol content of 0.0% by volume at a dosage of 1:1000. The term "successful" means that the addition of the final product produced an aroma profile characteristic of beer. The dosage is so low that the ethanol content in the final application is below 0.05% by volume, allowing it to be declared as "0.0% by volume" beer. Furthermore, the final product is clear and therefore readily applicable to typically clear beers such as pilsner.

[0096] The separated liquid phase obtained by the separation of the cyclodextrin-AOS complex in step (c) can, if necessary, be treated again with at least one cyclodextrin to maximize the yield and, in particular, recycled to step a).

[0097] In a further development of the invention, it is provided that the cyclodextrin-AOS complex separated in step (c) is diluted with water in step d1) to a desired final concentration of one or more flavorings or of cyclodextrin-AOS complex prior to the enzymatic treatment described in more detail below in step (d). In this way, the conditions for the most complete possible release of the extracted nonpolar organic substance(s) can be improved. Step (d)

[0098] The solid phase separated in step (c), which contains cyclodextrin-AOS complexes, is subjected in step (d) to an enzymatic treatment with at least one enzyme selected from the group comprising enzymes with amylase activity, preferably alpha-amylase, particularly preferably fungal-alpha-amylase, pulluanase and / or isoamylase, as well as mixtures of at least two of the aforementioned enzymes, to hydrolyze the cyclodextrin in these complexes.

[0099] The resulting mixture is optionally filtered.

[0100] In the targeted enzymatic treatment of host-guest complexes, the ring structure of the cyclodextrin is broken, and the previously complexed and optionally filtration-enriched target substances are released again without thermal energy. The available alpha, beta, gamma, and delta cyclodextrins and the complexes formed with these cyclodextrins as hosts can be partially and selectively hydrolyzed by various enzymes, particularly different amylases. This enables a process design that allows the cascaded release of nonpolar organic substances such as flavorings.

[0101] After step (d), a composition loaded with at least one flavoring substance is obtained. This composition also contains the breakdown products of cyclodextrin, such as mono-, di-, and oligosaccharides. These breakdown products are commonly found in foods and can therefore remain there or be removed as needed.

[0102] This enzymatic treatment according to step d) of the process according to the invention is carried out in the presence of water. In one embodiment of the invention, a step d1) is carried out before step d) in which the solid phase separated in step c) is mixed with water until the desired final concentration of one or more flavorings is reached. Preferably, a final concentration of at least 100 wt.% up to approximately 3750 wt.% water based on the total weight of the solid phase separated in step c) is used.

[0103] In one embodiment of the invention, the water added in step d1) has a temperature in the range of 4 to 80 °C, preferably in the range of 20 to 60 °C. The pH value can be adjusted to suit the enzyme used and is preferably in the range of pH 3.5 to pH 7.5, particularly preferably in the range of 4.5 or in the range of 5.2 to pH 5.6. The resulting aqueous mixture is then treated with at least one enzyme selected from the group comprising enzymes with amylase activity, preferably alpha-amylase, particularly preferably fungal-alpha-amylase, pulluanase and / or isoamylase, as well as mixtures of at least two of the aforementioned enzymes, preferably a mixture of two amylases, and enzymes that can degrade cyclodextrins as a side activity.

[0104] By using different enzymes, especially different amylases, the α-, β-, and / or γ-cyclodextrins used in the complexes can be partially and selectively hydrolyzed. This enables a process in which the flavor compounds are released in a cascade.

[0105] Alpha-amylase is particularly preferred, and treatment with Fungal-alpha-amylase is especially preferred. This Fungal-alpha-amylase is preferably used alone.

[0106] The fungal-alpha amylases used according to the invention are derived from microorganisms such as Aspergillus niger and Aspergillus oryzae. A suitable commercial product of fungal-alpha amylase is marketed under the name "Fungamyl®" by the company "Novozymes". Another suitable amylase is marketed under the name "Dextrozyme GA" by the company "Novozymes".

[0107] It is known that enzymes of different origins exhibit different reactivities. The required amount of enzyme for each mixture varies from mixture to mixture and from enzyme to enzyme. The preferred amount of enzyme used to hydrolyze a cyclodextrin in a cyclodextrin-flavoring complex depends essentially on the content of the solid phase containing the cyclodextrin-flavoring complex separated in step (c) and may also depend on the activity of the enzyme.

[0108] Within the scope of the invention, it was found that the preferred amount of the at least one enzyme is 5 to 1000 FAU per gram of the solid phase separated in step c). The unit "FAU" stands for "Fungal alpha-amylase Unit" and is a measure of the activity of an alpha-amylase, which is used by the company "Novozymes," for example, for the enzyme "Fungamyl®". More precisely, 1 FAU of an enzyme breaks down 5.26 g of starch in one hour under standard conditions (substrate: soluble starch, incubation time 7 to 20 min, temperature 37 °C, pH 4.7).

[0109] The treatment conditions, such as temperature and duration, can be varied within certain limits, particularly in step d), but temperatures between approximately 4 and 80 °C, preferably between approximately 20 and 60 °C, have proven advantageous, with treatment times of 0.5 to 50 hours being typical. The pH can be between 3.5 and 7.5, particularly preferably between 5.2 and 5.6. Incubation in a closed vessel is particularly preferred.

[0110] After enzymatic treatment, the maximum final cyclodextrin concentration is approximately 0.1 wt% based on the total weight of the aqueous mixture. The amount of cyclodextrin in the mixture is determined using established methods, namely HPLC.

[0111] Furthermore, additional enzymes can be used. For example, suitable enzymes, such as pulluanase and / or isoamylase, can be used prior to treatment with at least one amylase and / or in combination with at least one amylase.

[0112] In a further development of the invention, the enzyme is inactivated and / or separated from the composition containing the nonpolar organic substance extracted from the cyclodextrin-AOS complex by enzymatic treatment. Propylene glycol (PG) or glycerin, for example, can be used for this purpose.

[0113] The method according to the present invention provides a composition comprising at least one organic substance with at least one nonpolar residue selected from the group comprising volatile flavorings, non-volatile flavorings such as bitter substances, oil, fat and / or wax fractions, colorants, adhesives, in particular from plant and / or animal starting materials, as well as mixtures of at least two of the aforementioned substances, and at least one saccharide with a chain length of 6 or 7 or 8 or 9 glucose units, and optionally degradation products of at least one cyclodextrin, in particular glucose and / or maltose.

[0114] The composition can consist of at least one organic substance with at least one nonpolar residue and at least one saccharide with a chain length of 6 or 7 or 8 or 9 glucose units, as well as optionally degradation products of at least one cyclodextrin, in particular glucose and / or maltose.

[0115] In a preferred embodiment, the composition has an ethanol content of 0.0 vol.%.

[0116] The composition is preferably aqueous. Depending on its intended use, the composition can then be further processed. For example, at least one enzyme present in the composition can be inactivated by heating and / or by changing the pH value or by using, for example, propylene glycol (PG) or glycerol. Depending on the type of at least one nonpolar organic substance and the enzymes or organisms used to degrade the cyclodextrin, the nonpolar organic substance itself can also cause or support inactivation.

[0117] The degradation products of at least one cyclodextrin and the enzyme can be removed from the composition by common downstream processes such as liquid-liquid extraction, preserving the undiluted nonpolar organic substance. The resulting composition may then contain only at least one nonpolar organic substance, in particular at least one flavoring agent, which can be of either animal or plant origin. If the solubility of the nonpolar organic substance in water is exceeded, it separates as an oil phase or solid. In this way, the nonpolar organic substance can also be separated from the aqueous, sugar-containing phase.

[0118] Another possibility for separating the degradation products of at least one cyclodextrin is their oxidation to acids, which can then be removed by adsodium, anion exchangers or by forming chelates.

[0119] The invention thus provides a method for flavoring and / or stabilizing a product that is a food, beverage, cosmetic, or pharmaceutical product, in which the composition described above, or a composition produced by a method described above, is added to the product to be flavored. Such a product can then advantageously be marketed as "alcohol-free" (i.e., with an alcohol content of 0.0% by volume) and / or with the designation "FTNS." Depending on the organic substance obtained with at least one nonpolar residue, the invention provides not only a method for flavoring but also a method for stabilizing products, for example, using oil, fat, and / or wax fractions as a coating or coating component and / or with substances, such as phenolic substances, for color stabilization.

[0120] In the inventive process for flavoring and / or stabilizing a product, in particular a food, beverage, cosmetic or pharmaceutical product, in which the composition produced by the above-described process is brought into contact with the product to be flavored and / or stabilized, this "bringing into contact" can be carried out using any method known to the skilled person that is suitable for this purpose, preferably mixing.

[0121] The inventive process produces a food, beverage, cosmetic, or pharmaceutical product containing the inventive composition prepared by the process described above. A food, beverage, cosmetic, or pharmaceutical product that is particularly preferred is a drink, most preferably beer, and that can be marketed as "alcohol-free" (i.e., alcohol content at 0.0% by volume) and / or "FTNS".

[0122] The invention will be explained in more detail with reference to the accompanying figures and exemplary embodiments, without, however, being limited to the embodiment specifically described in each case. The invention also relates to all combinations of preferred embodiments, provided these are not mutually exclusive. The terms "approximately" or "about" in conjunction with a numerical value mean that values ​​at least 10% higher or lower, or 5% higher or lower, and in any case values ​​1% higher or lower, are included. The figures show: Figure 1 shows a schematic representation of the process for the selective separation of at least one organic substance with at least one nonpolar residue (AOS) in a first embodiment, using the example of the separation of an aroma substance from an aqueous solution, such as from a fermentation; Figure 2 shows a schematic representation of the process for the selective separation of at least one organic substance with at least one nonpolar residue (AOS) in a second embodiment, using the example of the separation of an aroma substance from an ethanolic extract, such as resulting from a SPE; and Figure 3 shows photographs of a comparative experiment on the sedimentation rate of gamma-cyclodextrin (left), beta-cyclodextrin aroma complex (center), and corresponding complexes in a mixture (right) of 10 wt% beta- and 90 wt% gamma-cyclodextrin based on the total mass of cyclodextrin.

[0123] In Figure 1Figure 1 is a schematic representation of an embodiment of the process for the selective separation of at least one organic substance with at least one nonpolar residue (AOS). First, in step (a), a starting mixture 10 is provided, which contains an aroma substance 1 in aqueous solution as a nonpolar organic substance. Such an aqueous aroma 3 can, for example, be a fermentate, which may have a low aroma substance concentration and optionally contains at least one solvent. In step (b), the starting mixture 10 consisting of aroma substance 1 in water 3 is brought into contact with at least one cyclodextrin 2. Water is already added as the solvent in step (a). The contact of the at least one cyclodextrin 2 with the at least one nonpolar organic substance 1 of the starting mixture 10 yields at least one cyclodextrin-AOS complex 12.Through complex formation, the aroma substance is extracted as a nonpolar organic substance from the solvent water 3 of the starting mixture 10. The complexes 12 are present in an aqueous phase.

[0124] In Figure 1 The continuation of the process is further shown, wherein step (c) involves the separation of the cyclodextrin-AOS complex 12 from the liquid phase by removing water 3. In step (d), the flavoring agent 1 is subsequently released from the cyclodextrin-AOS complex 12 by enzymatic treatment. This yields a composition 6 loaded with the nonpolar organic substance, in this case, flavoring agent 1. This composition comprises the concentrated flavoring 1 and, as a byproduct of the enzymatic treatment, saccharides 20.

[0125] In Figure 2Figure 1 is a schematic representation of a further embodiment of the process for the selective separation of at least one nonpolar organic substance (AOS). First, in step (a), a starting mixture 10 is provided, which, as an ethanolic extract, for example from a solid-phase extraction (SPE), contains a flavoring substance 1 in an ethanol-containing solution 4, which contains, for example, 80% by volume alcohol (ethanol). In step (b), the starting mixture 10, consisting of flavoring substance 1 in the ethanol-containing solution 4, is brought into contact with at least one cyclodextrin 2. The contact of the at least one cyclodextrin 2 with the at least one nonpolar organic substance 1 of the starting mixture 10 leads to the formation of at least one cyclodextrin-AOS complex 12. Through complex formation, the flavoring substance is extracted as a nonpolar organic substance from the alcohol-containing solution 4 of the starting mixture 10.Complexes 12 are present in an aqueous, alcohol-containing phase 4.

[0126] In Figure 2The continuation of the process is shown, with step (c) involving the separation of the cyclodextrin-AOS complex 12 from the liquid phase by removing ethanol 40. In step (d), the flavoring agent 1 is subsequently released from the cyclodextrin-AOS complex 12 by enzymatic treatment. For this purpose, the solid phase of the complex 12, consisting of cyclodextrin and flavoring agent separated in step (c), is added to water 3 to create suitable conditions for the enzymatic treatment. Step (d) yields a composition 6 loaded with the nonpolar organic substance, in this case, flavoring agent 1. This composition comprises the concentrated flavoring 1 and, as a byproduct of the enzymatic treatment, saccharides 20. Depending on the process and the ethanol concentrations that can be tolerated in the composition for the specific application, the composition can contain, for example, up to 10 vol% ethanol.

[0127] In Figure 3 Three photographs are shown from left to right. These depict an experimental setup consisting of three flasks. The flasks are filled with suspensions of cyclodextrin-AOS complexes. The flask on the left contains gamma-cyclodextrin complexes, the middle flask contains beta-cyclodextrin complexes, and the flask on the right contains a mixture (right) of 10 wt% beta-cyclodextrin and 90 wt% gamma-cyclodextrin, based on the total mass of cyclodextrin. In addition to these cyclodextrins, the suspensions shown contain ethanol-containing beer extract; each graduated cylinder contains 150 g of the beer extract and 9 g of either beta-cyclodextrin or gamma-cyclodextrin, or 9 g of the mixture of beta- and gamma-cyclodextrin. The suspensions were stirred for 48 hours at 6°C in the presence of the various cyclodextrins or the cyclodextrin mixture and then simultaneously transferred into the graduated cylinders.

[0128] After the start of the comparison experiment at time 0h (left image), the sedimentation of the particles from the suspensions can be observed over a period of 0.5h (middle image) up to a period of 1h (right image), whereby those from the beta-cyclodextrin-containing suspension have already sedimented after 0.5h and thus twice as fast as the particles from the gamma-cyclodextrin-containing suspension, in which a clear supernatant, visually distinguishable from the sediment at the bottom of the flask, only appears after 1h of experiment duration.

[0129] The suspension containing a mixture of only 10 wt% beta-cyclodextrin and 90 wt% gamma-cyclodextrin, based on the total mass of cyclodextrin, surprisingly exhibits the same optical sedimentation behavior as the suspension containing only beta-cyclodextrin. According to the invention, an addition of beta-cyclodextrin to gamma-cyclodextrin in a ratio of less than 1:1 is therefore sufficient to increase the settling rate of suspensions containing gamma-cyclodextrin to values ​​comparable to those of suspensions containing only beta-cyclodextrin. Examples of implementation: Example 1 for flavoring, e.g., 0.0% alcohol by volume beers

[0130] An ethanol-containing (70-80 vol%) solid-phase extract [1 kg with an aroma concentration of 4 g / L], containing isoamyl alcohol, isoamyl acetate, phenylethyl alcohol, hexanoic acid, ethyl hexanoate, and other aroma compounds, is mixed with 60 g of α-, β-, and / or γ-cyclodextrin, preferably with γ-cyclodextrin. The mixture is stirred for 48 h at 6 °C. The aroma compound-cyclodextrin complex is separated by filtration and dried. To maximize the yield, the ethanolic solid-phase extract can be mixed again with α-, β-, and / or γ-cyclodextrin, preferably with γ-cyclodextrin, at a rate of 30 g per kg of solid-phase extract. The resulting alcohol-free filter cake is dissolved in water and enzymatically treated to release the aroma compound.For this purpose, the aqueous mixture is treated with 1 µL / mL amylase (Fungamyl® from Novozymes) (1 microliter of amylase per 1 milliliter of aqueous mixture) and incubated for 48 h at 55 °C and a pH of 5.2 in a closed vessel (the maximum final cyclodextrin concentration is 0.1 wt% based on the total weight of the aqueous extract). The solubility of the extracted flavor components in water is low, therefore the resulting two-phase mixture can be homogenized with propylene glycol (up to 1:1 w / w) and subsequently filtered, if necessary.

[0131] The resulting alcohol-free, aroma-rich product (5 g / L) can now be used to flavor, for example, 0.0% alcohol by volume beers (dosage for application: 0.2:1000).

[0132] Within the scope of the invention, the content of cyclodextrin and in particular the final cyclodextrin concentration in a composition is determined by HPLC (detector: RI; separation column: Polysep GFC-P 2000 from Phenomenex; mobile phase: 10% methanol in water (isocratic); flow rate: 0.5 mL / min; pressure: 25 bar; oven temperature: 55 °C; run time: 30 minutes).

[0133] The concentration of nonpolar organic substance, for example the aroma concentration, of the composition according to the invention, in particular of an alcohol-free extract, is determined by GC-FID (2 g of sample were extracted with 2 g of cyclohexane. The organic phase is dried over Na₂SO₄, mixed with an internal standard and analyzed). Example 2 for flavoring e.g. 0.0% alcohol by volume beers

[0134] A ferment containing isoamyl acetate or 4-vinylguaiacol is treated with one molar equivalent of β-cyclodextrin (α- and / or γ-) with respect to the flavoring agent. To obtain a stable flavoring agent-cyclodextrin complex, the mixture is stirred for 48 h at 6 °C.

[0135] The flavoring agent-cyclodextrin complex is separated by filtration or concentrated on the retentate side.

[0136] The flavoring is released through enzymatic treatment of the flavoring-cyclodextrin complex. For this purpose, the aqueous mixture was treated with 1 µL / mL amylase (Fungamyl® from Novozymes) (1 microliter of amylase per 1 milliliter of aqueous mixture) and incubated for 48 h at 55 °C and pH 5.2 in a closed vessel (the maximum final cyclodextrin concentration is 0.1 wt% based on the total weight of the aqueous extract). The solubility of the extracted flavor components in water is low; therefore, the resulting two-phase mixture can be homogenized with propylene glycol (up to 1:1 w / w) and subsequently filtered, if necessary.

[0137] The resulting alcohol-free, aroma-rich product can now be used to flavor, for example, beers containing 0.0% alcohol by volume. The final cyclodextrin concentration is determined by HPLC (detector: RI; column: Phenomenex polysep GFC-P 2000; mobile phase: water (isocratic); flow rate: 0.8 mL / min; pressure: 20 bar; oven temperature: 55 °C; run time: 30 minutes). The aroma concentration of the alcohol-free extract is determined by GC-FID (2 g of sample were extracted with 2 g of cyclohexane. The organic phase is dried over Na₂SO₄, treated with an internal standard, and analyzed). Example 3 for flavoring 0.0% alcohol by volume beverages

[0138] An ethanol-containing (70-80 vol%) solid-phase extract (or a spirit or liqueur (15-96 vol)) containing isoamyl alcohol, isoamyl acetate, phenylethyl alcohol, and other flavorings was mixed with 60 g / L of a 1:9 beta- and gamma-cyclodextrin mixture. To obtain a sufficiently stable host-guest complex, the mixture was stirred overnight at 6 °C. The mixture was then allowed to settle for 2 hours (without beta-cyclodextrin, this process can take up to a day).

[0139] After sedimentation, the clear phase was removed and the sediment was dried by filtration.

[0140] To maximize yield, the aroma-depleted ethanolic clear solid phase extract can be treated again with a beta- and gamma-cyclodextrin mixture (1:9) and then left to settle, with the clear phase decreasing after sedimentation and subsequent filtration.

[0141] The resulting alcohol-free filter cake was dissolved in water and enzymatically treated to release the aroma. For this purpose, the aqueous mixture was treated with 1 µL / mL (microliters per milliliter) of Amylase Dextrozyme GA (Novozymes) and incubated for 48 h at 55 °C and pH 4.5 in a closed vessel. The resulting mixture was then filtered.

[0142] The resulting alcohol-free, aroma-rich product can now be used in low dosages, for example in the range of 0.01:1000 to 50:1000, to flavor beers containing, for example, 0.0% alcohol by volume. Example 4 for the extraction of apple wax-containing fraction from dried apple pomace

[0143] 1600 g of dry apple pomace was mixed with 8000 g of a 36 wt% aqueous cyclodextrin solution (beta-cyclodextrin:gamma-cyclodextrin in a 1:1 ratio, i.e., 18 g beta- and 18 g gamma-cyclodextrin per 1 L of water) and kneaded for 2 days at room temperature. The wet apple pomace was then filtered, and the cyclodextrin-apple wax complex-rich filtrate was collected as a suspension.

[0144] The suspension was centrifuged as step c) of the process according to the invention and the clear phase was decanted.

[0145] The extract obtained in this way (182.65 g) was mixed with 200 g of water and incubated for 48 h at pH 4.5 and 55°C with the amylase "Dextrozyme GA" (400 µL). The mixture was then filtered and dried to constant weight on a rotary evaporator. The resulting apple wax-rich fraction (16 g) was successfully tested as a component of a release agent for fruit gums. This means that the apple wax-rich fraction obtained in this way can be used as a release agent just as effectively as a release agent containing a wax (including apple wax) obtained by other known methods. Example 5 for the extraction of aromatic oil fraction from hop cones

[0146] 50 g of dry hop cones of the "Herkules" variety were mixed with 750 g of an 18 wt% aqueous solution of beta-cyclodextrin and kneaded for 2 days at room temperature. The wet hop cones were then filtered, and the cyclodextrin-oil complex-rich filtrate was collected as a suspension.

[0147] The suspension was centrifuged as step c) of the process according to the invention and the clear phase was decanted.

[0148] The resulting extract (28.99 g) was mixed with 15 g of water and incubated with the amylase "Dextrozyme GA" (88 µL) for 48 h at pH 4.5 and 55°C. The resulting aroma-rich extract has a high aroma load, allowing its use at a dosage of 1:1000 for flavoring, for example, non-alcoholic beer. Example 6 for the extraction of flavor compounds from ethanol-containing gentian root extract

[0149] An extract of gentian roots with an alcohol content of 60% by volume was mixed with 6% by weight of solid gamma-cyclodextrin. The mixture was stored overnight at 6°C. In this way, a sufficiently stable host-guest complex was obtained.

[0150] The host-guest complex was separated by filtration as step c) of the process according to the invention. The filter cake was mixed with water. The release of the flavor compounds was achieved by enzymatic treatment of the host-guest complex. For this purpose, the aqueous mixture was treated with 1 µL / mL of the amylase "Dextrozyme GA" (Novozymes) and incubated for 48 h at 55 °C in a closed vessel. The resulting mixture was then filtered.

[0151] The resulting aromatic product can now be used in low doses to flavor... e.g. used in products with an alcohol content of 0.0% vol. ethanol.

[0152] It is apparent to a person skilled in the art that the features of the individually illustrated examples can also be combined or interchanged. The scope of protection of the invention is defined by the accompanying claims. Reference symbol list

[0153] 1 Organic substance with at least one nonpolar residue, abbreviated: "nonpolar organic substance (AOS), flavoring, bittering agent, oil, fat, wax, adhesive, dye" 10 Starting mixture 2 Cyclodextrin 12 Guest-host complex of cyclodextrin and organic substance with at least one nonpolar residue (AOS), cyclodextrin-AOS complex 20 Saccharides 3 Water 4 Ethanol-containing solution 40 Ethanol 5 Enzyme 6 Composition

Claims

1. A method for flavoring and / or stabilizing a product which constitutes a food product, luxury food product, cosmetic product, or pharmaceutical product, comprising adding, to the product to be flavored and / or stabilized, a composition (6); wherein the composition (6) is produced by a method for selectively separating at least one organic substance comprising at least one apolar group (AOS) (1) and / or at least one or more aromatic substances comprising: (a) providing a starting mixture (10) which contains at least one organic substance comprising at least one apolar group (AOS) and / or at least one or more aromatic substances (1) and optionally at least one solvent (3; 4; 40); (b) bringing the starting mixture (10) into contact with at least one cyclodextrin (2); wherein the at least one organic substance (1) comprising at least one apolar group is selected from the group comprising volatile aromatic substances, non-volatile aromatic substances such as, e.g., bitter substances, oil, fat and / or wax fractions, colorants and adhesives, in particular from plant-based and / or animal-based starting materials, and mixtures of at least two of the substances mentioned, and wherein at least one solvent, in particular water, is added in step (a) and / or in step (b) and / or following step (b); and wherein, as a result of the contacting of the at least one cyclodextrin (2) with the at least one organic substance comprising at least one apolar group and / or the at least one or more aromatic substances (1) of the starting mixture (10), at least one cyclodextrin-AOS complex and / or cyclodextrin-aromatic substance complex (12) is obtained in a liquid, in particular aqueous, phase (3; 4); comprising the further steps of (c) separating the cyclodextrin-AOS complex and / or the cyclodextrin-aromatic substance complex (12) from the liquid phase; and (d) enzymatically treating the separated cyclodextrin-AOS complex and / or the cyclodextrin-aromatic substance complex (12) to obtain a composition (6) that is loaded with at least one organic substance comprising at least one apolar group (AOS) (1) and / or with at least one aromatic substance; wherein the enzymatic treatment is performed using at least one enzyme selected from the group comprising enzymes with amylase activity, preferably alpha-amylase, most preferably with fungal alpha-amylase, pulluanase and / or isoamylase, and mixtures of at least two of these enzymes.

2. The method of claim 1, characterized in that in step (a) and / or in step (b) and / or following step (b), a water content in the range from 15 vol% to 35 vol%, preferably from 20 vol% to 30 vol%, is adjusted; and / or an ethanol content is adjusted so as to be at least 40 vol%, preferably to an ethanol content in the range from 40 vol% to 60 vol%.

3. The method of claim 1 or 2, comprising a further step of (d1) diluting the cyclodextrin-AOS complex separated in step c) and / or the cyclodextrin-aromatic substance complex (12) with water prior to the enzymatic treatment in step d).

4. The method as claimed in any of claims 1 to 3, characterized in that step (d) is performed such that the cyclodextrin concentration in the composition (6) that is loaded with at least one apolar organic substance (AOS) and / or with at least one aromatic substance (1) is less than 0.5 wt%, preferably less than 0.1 wt%.

5. The method as claimed in any of claims 1 to 4, comprising a further step of (e) filtering the mixture resulting from the in particular enzymatic treatment of the cyclodextrin-AOS complex and / or the cyclodextrin-aromatic substance complex (12), wherein a composition (6) loaded with at least one organic substance comprising at least one apolar group (AOS) and / or with at least one aromatic substance is obtained.

6. The method as claimed in any of claims 1 to 5, wherein in step (a) and / or in step (b) and / or following step (b) at least one solvent is used, which solvent is selected from the group comprising water, C1 - C4 alcohols, diethyl ether, acetone, or mixtures thereof, preferably water, ethanol and mixtures thereof.

7. The method as claimed in any of claims 1 to 6, wherein as a cyclodextrin (2), at least one substituted or unsubstituted α-cyclodextrin, at least one substituted or unsubstituted β-cyclodextrin, at least one substituted or unsubstituted γ-cyclodextrin, at least one substituted or unsubstituted δ-cyclodextrin, or mixtures thereof, preferably substituted or unsubstituted γ-cyclodextrin, most preferably a mixture consisting of 10 wt% of substituted or unsubstituted β-cyclodextrin and 90 wt% of substituted or unsubstituted γ-cyclodextrin is used.

8. The method as claimed in any of claims 1 to 7, wherein the separating of the cyclodextrin-AOS complex (12) in step (c) is carried out by centrifugation or filtration, preferably by vacuum filtration or nanofiltration.

9. The method as claimed in any of claims 1 to 8, comprising a further step of (c1) allowing to rest, prior to the separating of the cyclodextrin-AOS complex and / or the cyclodextrin-aromatic substance complex from the liquid phase; wherein the resting phase according to step (c1) is performed in particular over a duration of up to 24 hours, preferably over a duration of up to 12 hours, most preferably over a duration of 2 hours.

10. The method as claimed in any of claims 1 to 9, wherein the separated liquid phase as obtained in step (c) by separating the cyclodextrin-AOS complex and / or the cyclodextrin-aromatic substance complex (12) is admixed again with at least one cyclodextrin (2), and in particular is recirculated to step (a), to maximize yield.

11. The method as claimed in any of claims 1 to 10, wherein the cyclodextrin-AOS complex and / or the cyclodextrin-aromatic substance complex (12) separated in step (c) is prior to the in particular enzymatic treatment in step (d), diluted in a step (d1) with water, so as to obtain a desired final concentration of one or more hydrophilic organic substances and / or one or more aromatic substances (1).

12. The method as claimed in any of claims 1 to 11, wherein the at least one enzyme (5) is used in an amount ranging from 5 FAU to 1000 FAU per gram of the separated solid phase.

13. The method as claimed in any of claims 1 to 12, comprising introducing at least one aromatic substance and / or at least one organic substance (1) comprising at least one apolar group into a food or a luxury food or a beverage or a cosmetic product or a pharmaceutical product with an ethanol content of 0.0 vol%.