Method for selectively separating at least one organic substance containing at least one non-polar group and use of said substance in food, luxury goods, cosmetics or pharmaceuticals

Abstract

The present invention relates to a method for selectively separating at least one non-polar organic substance, comprising the steps of: (a) providing a starting mixture 10 containing at least one organic substance 1 comprising at least one non-polar group and, optionally, a solvent 3; 4; 40; (b) contacting the starting mixture with at least one cyclodextrin 2, thereby obtaining a cyclodextrin-aroma complex 12; optionally, (c) separating the complex 12 from the liquid phase; (d) treating the separated complex 12, in particular with an enzyme; and, optionally, filtering the resulting mixture, to obtain a composition 6 loaded with at least one organic substance comprising at least one non-polar group. The present invention also relates to the composition 6 loaded with at least one organic substance 1 comprising at least one non-polar group, and to the use of said composition 6 for incorporating at least one non-polar organic substance 1 into a food, luxury product, cosmetic, or pharmaceutical product.

Description

[Technical Field]

[0001] The present invention relates to a method for selectively separating organic substances comprising at least one non-polar group, to a composition comprising at least one organic substance comprising at least one non-polar group, and to the use of this selectively separated organic substance in foods, luxury goods, cosmetics or pharmaceuticals. [Background technology]

[0002] The growing interest of people in a healthy lifestyle has promoted the sale of non-alcoholic beverages such as sparkling wine (champagne), lemonade or beer, making it necessary to explore new production technologies that not only have an enjoyable taste and a pleasant smell, but also allow the production of beverages and foods that do not use any ethanol at all.

[0003] According to the European Food Information Regulation (LMIV), alcohol labeling is only mandatory from 1.2 vol%. The term "alcohol-free" does not necessarily mean "free of alcohol." For example, beer and wine advertised as "alcohol-free" may contain up to 0.5 vol% alcohol. If another beverage (other than wine or beer) is advertised as "alcohol-free," it must contain no alcohol at all, i.e., the alcohol content is 0.0 vol%. Otherwise, the "alcohol-free" labeling is considered misleading and deceptive to consumers. As used herein, the term "alcohol" refers to ethanol.

[0004] Important ingredients in foods, luxury goods, cosmetics, and pharmaceuticals are obtained by solvent extraction processes, often using ethanol. However, ingredients obtained in this way may contain ethanol residues, and products containing such ingredients are not "alcohol-free."

[0005] Components that are of interest in the present context and that are obtained by extraction in ethanol include, in addition to volatile aroma substances, non-volatile aroma substances, such as bitter substances, oil, fat and / or wax fractions, colorants and adhesives, aroma substances from plant and / or animal starting materials and other natural sources, such as microbial or fungal origin, and / or of synthetic origin.

[0006] These substances can be either non-polar or amphiphilic, as they have at least one non-polar group in the molecule, and in the following they will be collectively and succinctly encompassed by the term "non-polar organic substances", sometimes abbreviated as "AOS" for ease of reading.

[0007] For many years, there has been great interest in the recovery of aroma substances, in particular from the dealcoholization and / or concentration processes of products such as beer / wine or from juices, or in the removal of undesirable substances such as bitter substances from citrus juices, but also in the recovery or removal of non-polar organic substances from cosmetics and pharmaceuticals, and the extracts thus obtained can be used for the alcohol-free re-aromatization, i.e. flavoring, of products or in other ways, for example as coating agents in the case of waxes.

[0008] The literature describes four basic separation processes for the recovery of desired apolar organic substances and the removal of undesired apolar organic substances, in particular aroma substances soluble in liquids such as water or ethanol: (1) liquid / liquid extraction with organic solvents, which ultimately yields the pure substance or the desired fraction by evaporation of the solvent used; (2) rectification of the liquid containing the AOS or aroma substances; (3) sorption processes, which enrich the apolar aroma substances from the liquid phase into the solid phase; (4) precipitation and crystallization of solid aroma components; and (5) direct extraction with supercritical gases, percolation. For fat, oil and wax fractions from plant or animal starting materials, mechanical processes such as steam distillation and expression are also applicable.

[0009] Although extraction has widespread applications, the use of large amounts of organic solvents, often chlorinated, is highly problematic from an environmental and sustainability perspective. Furthermore, thermally unstable substances may be irreversibly destroyed upon evaporation of the solvent. Recovery of aroma substances from ethanolic extracts, for example by liquid / liquid extraction, is also difficult, since ethanol itself is soluble in various organic solvents. Direct re-aromatization using ethanolic extracts is ruled out, as the final product would not meet the description "alcohol-free" (i.e., 0.0 vol% alcohol content).

[0010] Usually, it is not possible to selectively remove ethanol or water by rectification, as azeotropes are formed with the aroma substances contained in the extract, and thermally unstable aroma-imparting components may be irreversibly destroyed.

[0011] In the case of sorption processes, the desired enriched material is re-eluted with a non-polar medium. Especially in the food industry, ethanol is often used as an eluent, which makes the extract unusable for applications with 0.0 vol%. Carbon dioxide is also used as an additional eluent. Patent Document 1 describes a process for extracting two or more fractions from hop oil by first treating the hop oil supported on an adsorbent carrier with liquid carbon dioxide to separate a first fraction, and then treating it with supercritical carbon dioxide to separate a second fraction. By combining supercritical carbon dioxide with a co-solvent, other fractions can be separated. Elution with liquid / supercritical carbon dioxide avoids the previously mentioned undesirable use of alcohol, but results in high yield losses when venting gas. The construction and operation of CO2 extraction plants are expensive and have specific safety requirements.

[0012] Mechanical processes such as pressing are mainly used in the field of oil extraction and do not require any solvents, however, these processes cannot be universally applied to the recovery of all non-polar organic substances, especially all oils and waxes from plants.

[0013] It is therefore an object to provide a method for selectively obtaining at least one non-polar organic substance, in particular at least one aroma substance, preferably from natural sources such as plant or animal products, so that the obtained substance can be used in food, luxury goods, cosmetics and / or pharmaceuticals that can be labelled as "alcohol-free". Such products must in particular meet the quality requirements for the labelling "FTNS" ("From the named source"). The aim is to enable so-called "clean label" products. A further object of the present invention is to make it possible to selectively obtain non-volatile aroma substances, such as bitter substances, as well as to selectively obtain oil, fat and / or wax fractions from plant and / or animal starting materials.

[0014] The prior art is known to employ numerous processes that use cyclodextrins to remove undesirable substances from food systems. Cyclodextrins are water-soluble and toxicologically and environmentally harmless. For example, cyclodextrins are used to remove cholesterol from butter (Patent Document 2) or eggs (Patent Documents 3 and 4). Such removal processes typically involve two stages: first, mixing cyclodextrin with the food system to form a complex between the cyclodextrin and the undesirable substance (guest-host complex), and then removing this complex from the food system. The cyclodextrin can be released again from the complex (decomplexation), for example, by treatment with hot water or alcohol (40°C to 100°C), and can then be reused in the removal process.

[0015] The previously known use of cyclodextrins in the extraction of aroma substances has the disadvantage that larger amounts of solvent are used for decomplexation and that heating is required, which leads to a dilution of the aroma substance concentration and also causes additional technical complications and high costs.When ethanol is used as the solvent for decomplexation, in addition to the dilution of the aroma substance concentration, another problem arises, which is related to the fact that this ethanol cannot be removed again without affecting the flavor profile.

[0016] Surprisingly, the inventors have found that cyclodextrins in aqueous and alcoholic media are capable of reversibly incorporating into their cavities such ingredients, in particular food ingredients, such as aromatic substances, bitter substances, oils, fats and waxes, adhesives, and colorants, which can be obtained by extraction in ethanol.

[0017] Such components have at least one non-polar group, i.e., they can be either non-polar or amphiphilic. Amphiphilic components have at least one polar moiety in addition to at least one non-polar moiety. Organic substances containing at least one non-polar group, i.e., they also have a polar moiety, and are particularly amphiphilic, will be referred to below as "non-polar organic substances." This term may also be abbreviated as "AOS." Organic substances containing at least one non-polar group within the meaning of the present invention, obtained by extraction in ethanol, are more hydrophobic in water than ethanol. [Prior art documents] [Patent documents]

[0018] [Patent Document 1] European Patent No. 3063260 [Patent Document 2] Australian Patent Application Publication No. 638531 [Patent Document 3] European Patent Application Publication No. 326469 [Patent Document 4] European Patent No. 475451 Summary of the Invention

[0019] Therefore, the present invention provides the possibility of selectively transferring a non-polar organic substance from a starting mixture to a complex consisting of a cyclodextrin and the non-polar organic substance by selecting a suitable cyclodextrin.

[0020] The present invention therefore achieves the above-mentioned object in a very simple manner by means of a method according to claim 1 and a composition according to claim 17.

[0021] The present invention provides a method for selectively separating at least one organic substance (AOS) containing at least one apolar group and / or at least one or more aromatic substances, comprising: (a) providing a starting mixture containing at least one organic substance (AOS) containing at least one apolar group and / or at least one or more aromatic substances, and optionally at least one solvent; (b) contacting the starting mixture with at least one cyclodextrin; wherein in step (a) and / or step (b) and / or after step (b) at least one solvent, in particular water, is added, The method provides that, as a result of contacting at least one cyclodextrin with at least one organic substance containing at least one non-polar group and / or at least one or more aromatic substances of the starting mixture, at least one cyclodextrin-AOS complex and / or at least one cyclodextrin-aromatic substance complex is obtained in a liquid phase, in particular an aqueous phase.

[0022] The terms "the cyclodextrin" or "cyclodextrin" are understood to mean the entire cyclodextrin molecule in the respective material system, for example the starting mixture or the liquid phase. This also applies correspondingly to the expressions "the cyclodextrin-AOS complex" or "cyclodextrin-AOS complex".

[0023] The process according to the invention makes it possible to obtain compositions loaded with at least one organic substance comprising at least one apolar group, such as an aromatic substance or a bitter substance, which provides for example a reversible protection of thermally unstable substances, which can be removed as soon as they are no longer needed.

[0024] Depending on the material system in which at least one non-polar organic substance is present in the starting mixture, it may not be necessary to add any solvent or further solvent to be able to carry out the method of the present invention. For example, the starting mixture may already be a solution itself, or may contain sufficient solvent in the form of water after the addition of the cyclodextrin dissolved in water in step (b).

[0025] The quality of complexation can be influenced by adjusting the solvent content of the starting mixture, in particular the water content and / or the ethanol content. Thus, the present invention offers the option of adding at least one solvent in step (a) and / or step (b) and / or after step (b), depending on the application. In this case, a person skilled in the art will adjust the solvent content so that the quality of complexation is optimized for the application. The choice of cyclodextrin, the temperature, and / or the energy input during complexation, for example by adjusting the agitator speed, also offer the possibility of influencing the quality of complexation.

[0026] The complexation conditions can be adapted to the non-polar organic substances to be separated within the scope of the present invention. In particular, it is possible to adjust the extraction conditions within the scope of the present invention so that, for example, bittering agents can be selectively complexed and isolated or left intact.

[0027] In an advantageous embodiment of the invention, it is provided for this purpose to adjust, in step (a) and / or step (b) and / or after step (b), the water content to be in the range of 15% to 35% by volume, preferably 20% to 30% by volume, in particular by adding water, and / or to adjust the ethanol content to be in the range of at least 40% by volume, preferably 40% to 60% by volume, in particular by adding ethanol.

[0028] Ethanol-containing extracts of non-polar organic substances, especially aromatic and / or bitter substances, are highly suitable for use in flavorings added to foods, luxury goods, cosmetics or pharmaceuticals, as they have a natural, complex flavor profile very similar to that of the starting material. In the present invention, this complexity can be obtained after dealcoholization for alcohol-free products by adjusting the water and / or ethanol content during complexation of the non-polar organic substances with cyclodextrins.

[0029] During process optimization, it was found that dilution with water to a range of 15% to 35% by volume, preferably 20% to 30% by volume, significantly improves the quality of cyclodextrin complexation with volatile aroma or flavor components. However, depending on the substance system, the addition of water surprisingly results in the inability to further complex non-volatile aroma or flavor components, such as bitter substances, thereby altering the natural aroma profile of the extract. Surprisingly, in the context of the present invention, it was found that complexation of bitter substances can still be achieved if the ethanol content of the liquid phase during complexation with cyclodextrin is at least 40% by volume.

[0030] The present invention therefore makes it possible to obtain a characteristic aroma or flavor profile from an ethanol extract which contains, in addition to volatile aroma substances, also non-volatile aroma substances, such as bitter substances. The present invention therefore also offers the possibility of selectively removing non-volatile aroma substances, such as bitter substances, from extracts containing non-polar organic substances, i.e., under conditions unfavorable to their complexation, in particular at an ethanol content of less than 40 vol.%.

[0031] Although the use of cyclodextrins (CDs) in food systems is known, to the inventors' knowledge they have never been used to selectively obtain aroma substances or other organic substances bearing at least one apolar group. Therefore, the present invention provides a method for using cyclodextrins to obtain alcohol-free (i.e., ethanol-free), non-declarable FTNS ("from named source") aroma substances or bitter substances not only from aqueous extracts but also (especially) from ethanolic extracts.

[0032] The present invention therefore provides a method that allows the separation / obtaining in concentrated form, using cyclodextrins, of organic substances comprising at least one apolar group and / or at least one fragrance substance or substances from aqueous, ethanolic or other extracts, while reducing the technical complexity (and solvent consumption) and allowing the entire process to be carried out at moderate temperatures. The apolar organic substances, such as fragrance substances, thus obtained are suitable for use in foods, luxury goods, cosmetics and / or pharmaceuticals that may be labeled as "alcohol-free."

[0033] According to the present invention, a process has been found that also makes it possible to selectively obtain oil, fat and / or wax fractions, especially from plant and / or animal starting materials. Cyclodextrins have proven to be suitable adjuvants for this purpose in a surprisingly simple manner, since they are water-soluble and can bind to target substances. The target substances can be removed again from the cyclodextrin-AOS complex without the need for further solvents.

[0034] The present invention allows extraction from a starting mixture with cyclodextrin to obtain all organic substances containing at least one nonpolar group that are sufficiently hydrophobic to react and complex with the host α- and / or β- and / or γ- and / or δ-cyclodextrin as guests. In other words, each organic substance containing at least one nonpolar group is sufficiently hydrophobic to enter the cavity of α- and / or β- and / or γ- and / or δ-cyclodextrin from solution. In the context of the present invention, it is not necessary for the entire nonpolar substance to be complexed within the cyclodextrin cavity. For example, in the case of fats and organic acids, essentially only the hydrophobic portion of the substance is complexed within the cyclodextrin cavity. According to the model concept, the polar portion of the substance protrudes from the cavity. In the case of amphiphilic substances, the hydrophobic portion is also accommodated within the cavity, while the hydrophilic portion remains untouched.

[0035] The process according to the invention avoids exposing the non-polar organic material to high thermal loads. For this purpose, the invention provides for carrying out the process at a maximum temperature ranging from at least 40°C to at most 70°C, preferably at least 40°C to no more than 55°C. These temperatures are particularly relevant for step (d), which will be explained further below. In step (b), the temperature is preferably lower, in particular in the range of 4°C to 10°C, preferably 6°C.

[0036] The present invention thereby provides a method for preparing a polymerizable polymer comprising: secondary metabolites, phytochemicals, especially secondary plant substances, Volatile aromatic substances, non-volatile aromatic substances, e.g. bitter substances, coloring agents, an oil fraction, a fat fraction and / or a wax fraction, glue, in particular of natural origin, preferably from plant and / or animal starting materials, as well as mixtures of at least two of the substances mentioned, This gives the advantage that the compound can be used in a variety of fields of application.

[0037] Of particular interest herein are natural fragrance substances, including those obtained from microorganisms or fungi. Phenolic substances can be important in stabilizing colorants.

[0038] In an advantageous embodiment of the invention, the method comprises the steps of: (c) a further step of separating the cyclodextrin-AOS complex and / or the cyclodextrin-aroma complex from the liquid phase, in particular the solvent, Includes.

[0039] This concentrates the (still complexed) non-polar organic substances, in particular aromatic substances. The cyclodextrin-AOS complex and / or cyclodextrin-aroma complex is separated from the liquid phase to obtain a separate solid phase containing at least the cyclodextrin-AOS complex and / or cyclodextrin-aroma complex. The liquid phase contains water and a solvent, in particular ethanol, which was provided in the starting mixture together with the non-polar organic substance, such as at least one aromatic substance, and / or which was added in the process according to the invention.

[0040] The substantially solvent-free solid is retained after step (c). "Substantially solvent-free" particularly means that the solvent, e.g., water and / or ethanol, may be adsorbed on the solid phase, but is no longer freely present in the solid phase. Depending on the application, the separation of water in step (c) can also separate undesired components of the starting mixture from the filter cake, equivalent to the function of a washing step.

[0041] The separation of not only water but also other liquids from the solid phase, particularly filter cake, can be supported by drying within the scope of the present invention.In a simple embodiment, compressed air can be passed through the separated solid phase, for example, in the form of filter cake, towards the filter layer, particularly at a pressure of 2 bar.Inert gas can also be used instead of compressed air.

[0042] The known methods of removing alcohol using evaporators or reverse osmosis have the drawback that other important, especially volatile aroma substances that are crucial to the characteristic taste of the extract, are also removed together with the ethanol. On the other hand, thermally unstable substances may be destroyed or undesirable changes in taste may occur. The method described in the present invention allows for highly selective removal of ethanol from the extract. By equally complexing both volatile and non-volatile aroma components, the complexity of the extract is maintained.

[0043] In a further advantageous embodiment of the invention, the method comprises the steps of: (d) a further step of treating the specifically isolated cyclodextrin-AOS complex and / or cyclodextrin-fragrant substance complex with an enzyme and / or with an organism selected from the group comprising yeasts, fungi and mixtures thereof, capable of degrading cyclodextrins, to obtain a composition loaded with at least one organic substance comprising at least one apolar group (AOS) and / or at least one fragrance substance, Includes.

[0044] Surprisingly, it has been found that the enzymatic treatment not only degrades free cyclodextrin, but also cyclodextrin bound to at least one non-polar organic substance, such as a fragrance substance, in a complex, thereby enriching (without damaging) the non-polar organic substance, e.g., the fragrance substance, in the resulting composition.

[0045] In addition to or instead of the enzymatic treatment described in more detail further below, this degradation of the cyclodextrins and thus the release of the non-polar organic substances from the complexes can also be achieved within the scope of the present invention by using organisms from the group comprising yeasts, fungi and mixtures thereof that are capable of degrading cyclodextrins.

[0046] The treatment of the cyclodextrin-AOS complex and / or cyclodextrin-fragrant substance complex to obtain non-polar organic substances comprises: (d1) a further step of diluting the cyclodextrin-AOS complex and / or cyclodextrin-aroma complex separated in step (c) with water, especially before the enzymatic treatment in step (d), can be supported by

[0047] In particular in the case of the preceding step (c) for the separation of water as described above, the water added in step (d1) does not contain any constituents that are undesirable for enzymation.

[0048] In a further advantageous embodiment of the present invention, the pH value for the enzymatic treatment can be adjusted in coordination with the enzyme used, for example to a value in the acidic range, in particular to pH 4.5. Within the scope of the present invention, the activity of the enzyme can be influenced by adjusting the pH value. When using various cyclodextrins, this effect can be utilized to first decompose one type of cyclodextrin by adjusting the pH, and then, by changing the pH, decompose another type of cyclodextrin using the same enzyme.

[0049] The advantages of the method according to the invention will now be explained using fragrances as an example of non-polar organic substances. These advantages also apply to the other non-polar organic substances mentioned above. The method of the invention makes it possible to obtain a composition in which fragrances are present in concentrated form from a diluted starting mixture. The separation can be carried out gently and completely. Furthermore, this method makes it possible to obtain, from a starting mixture that may have a very high alcohol content (i.e., up to 80 vol%), a fragrance-loaded composition that can be used in foods, luxury goods, cosmetics and / or pharmaceuticals that can be labeled as "alcohol-free."

[0050] The process according to the invention can be carried out at temperatures below 55°C or even below room temperature, making it possible to obtain highly volatile and moderately volatile (i.e., with an evaporation number below 10 or between 35 and 10, respectively) and / or thermally unstable aroma substances. 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 cascade release of aroma substances. These points are discussed in more detail further below.

[0051] According to a further embodiment of the present invention, it is contemplated that step (d) is carried out so that the cyclodextrin concentration in the at least one non-polar organic substance (AOS), in particular the at least one loaded composition, is less than 0.5 wt %, preferably less than 0.1 wt %, such that the low cyclodextrin content does not affect the use of the resulting non-polar organic substance (AOS).

[0052] The cyclodextrin concentration can be adjusted in relation to the total mass of the final composition in selected embodiments of the method according to the invention. For example, after steps (a), (b), (c), and (d) are performed on an aqueous fragrance solution, the composition can consist of water, fragrance, cyclodextrin, and enzyme degradation products, with residual cyclodextrin at a content of less than 0.1 wt%. If components such as water and enzyme are removed in further process steps, the enzymatic treatment can further reduce the cyclodextrin concentration, thus resulting in a water- and enzyme-free product with a content of less than 0.1 wt%.

[0053] The adjustment of the cyclodextrin concentration can be facilitated by removing components that are not desired in the final product, which may include undegraded cyclodextrin-AOS complexes. To this end, according to a further advantageous embodiment of the invention, the method comprises: (e) a further step of filtering the mixture resulting from the treatment, in particular with an enzyme, of the cyclodextrin-AOS complex and / or cyclodextrin-aroma complex to obtain a composition loaded with at least one organic substance comprising at least one apolar group (AOS), Includes.

[0054] Process (a) In the present invention, "fragrant substances" refer to organic substances that are volatile, especially at room temperature, and induce or alter the perception of smell and taste. Fragrant substances are often alcohols, acids, esters, lactones, aldehydes, ketones, acetals, ketals, ethers, epoxides, and their analogous sulfur compounds; oxygen heterocycles, nitrogen heterocycles, and sulfur heterocycles, heteroaromatic compounds (e.g., alkylpyrazines), amines, and amides; simple or complex saturated and unsaturated aliphatic and alicyclic compounds, aromatic compounds, and terpenes. The main types of fragrances are natural, nature-identical, and artificial fragrances. In the context of the present invention, so-called natural fragrances, whose starting materials are plant or animal origin, are preferred. Animal-derived starting materials include, for example, honey, milk, meat, bones, and body fluids. Starting materials of plant origin include plants or plant parts such as flowers, buds, leaves, stems, stalks, bark, roots, tubers, bulbs, rhizomes, fruits, nuts, berries, seeds, fruits, and vegetables. These starting materials can be provided, for example, in fresh, cooked, germinated, dried, or fermented form, or in a form prepared for consumption as a food or beverage (e.g., beer, wine, sparkling wine, or spirits such as whiskey). These starting materials can be used directly as a starting mixture in the method according to the present invention, or they can be treated by at least one process known to those skilled in the art before being used as a starting mixture. Such processes include, for example, dissolving, dispersing, purifying, mashing, maceration, fermentation, and / or separation processes, such as extraction and filtration. Examples of plants that yield ethanolic extracts containing bitter substances are gentian, chiretta, and wormwood.

[0055] In addition to such "aromatics," the starting mixture may contain other constituents of starting materials of plant or animal origin. In particular, the starting mixture may contain one or more non-polar organic substances, including non-volatile aroma substances, such as bitter substances, oil fractions, fat fractions and / or wax fractions, especially from plant and / or animal starting materials, as well as mixtures of at least two of the substances mentioned.

[0056] Within the scope of the present invention, the starting mixture can be prepared in the form of a solid or liquid dispersion, particularly a powder or a solution and / or suspension and / or emulsion. In principle, the starting mixture can contain any solvent in which the nonpolar organic substance to be separated dissolves and can be easily accommodated in or displaced from the cavity of the cyclodextrin molecule containing at least one substance to be separated. Solvents that can be used within the scope of the present invention in step (a) and / or step (b) and / or after step (b) include, for example, water, C1-C4 alcohols, diethyl ether, acetone, etc., or mixtures thereof. It is preferred to use a solvent selected from the group consisting of water, ethanol, or mixtures thereof. It is also possible to add a solvent during the preparation of the starting mixture in step (a). For example, a starting mixture containing ethanol and / or water can be directly extracted with the cyclodextrin according to the present invention, depending on the substance to be separated.

[0057] In one embodiment of the present invention, at least one of the above-mentioned solvents may be added not only in step (a) but also in step (b).

[0058] The preferred solvent content is less than 100 wt%. A person skilled in the art will select the solvent content and the mixing ratio of two or more solvents depending on the solubility of the fragrant substance(s) to be separated, the efficiency of complex formation, and also taking into account the economics of the process. A starting mixture in which at least one non-polar organic substance, in particular one or more fragrant substances, is dissolved in a minimum amount of solvent is particularly preferred.

[0059] The higher the solvent content, the higher the solubility of the fragrance substance to be separated and the cyclodextrin used, and the faster the rate of complex formation. However, excessively dilute solutions must be avoided, as this makes the process uneconomical and also reduces the probability of contact and interaction between the fragrance molecules and the cyclodextrin. According to one model concept, these relationships arise from the fact that as the concentration of a non-polar organic substance increases, the complex between this substance and the cyclodextrin is preferentially formed due to a driving gradient. In the case of excessive dilution, the driving force for complex formation becomes insufficient.

[0060] The starting material is preferably in a form prepared for consumption as a food or beverage (e.g., beer, wine, sparkling wine, spirits, etc.), which is subjected to extraction, preferably solid-phase extraction (abbreviated as SPE, formerly known as "sorbent extraction"), and the resulting solvent-containing solid extract is used as the starting mixture. Ethanol-containing solid extracts are particularly preferred, and the ethanol content can be up to 80 vol% based on the total volume of the solid extract. The content of the aromatic substance(s) is at least about 0.1 wt%, preferably 0.5 wt% to 8 wt%, based on the total weight of the solid extract.

[0061] Any kind of seeds, especially cereal grains and products obtained from cereal grains, such as malt, wort, mash, beer, etc., are other preferred starting materials. The cereal grains can be selected from the group including, for example, barley, wheat, rye, spelt, corn, oats, rice, millet, triticale, and mixtures thereof. This starting material can be used directly as a starting mixture in the method according to the invention or can be processed by at least one process known to those skilled in the art (see above). Beer wort and / or mash is particularly preferred as starting material, which is subjected to fermentation and the resulting fermented liquid is used as the starting mixture. The content of the aromatic substance(s) is at least about 0.01 wt %, preferably 0.1 wt % to 8 wt %, based on the total weight of the fermented liquid.

[0062] Process (b) In step (b) of the process according to the invention, the cyclodextrin-AOS complex is formed by contacting at least one cyclodextrin with the starting mixture. The components can be mixed in powder form or in suspension and / or emulsion and / or solution.

[0063] Various processes for generating guest-host complexes are known, with preparation in a solvent (the "slurry method", which is co-precipitation and complex formation in suspension) or kneading methods (see, for example, SK et al. / / Research Journal of Pharmaceutical, Biological and Chemical Sciences., 2013, Vol. 4, No. 2, pp. 1694-1720) being the most commonly applied.

[0064] In the present invention, co-precipitation or complexation in suspension ("slurry method") is used, in which at least one cyclodextrin is added to an aqueous or ethanol-containing solution or suspension of the starting mixture and the precipitated complex is isolated.

[0065] The "cyclodextrin" that can be used includes substituted or unsubstituted α-cyclodextrin, at least one substituted or unsubstituted β-cyclodextrin, at least one substituted or unsubstituted γ-cyclodextrin, or at least one substituted or unsubstituted δ-cyclodextrin, or mixtures thereof, preferably substituted or unsubstituted γ-cyclodextrin. The use of unsubstituted cyclodextrin is preferred in such cases, since the use of substituted cyclodextrins can result in substances that may be undesirable in foods as degradation products of the optional process step (d) of the enzymatic treatment and that may be removed in further separation steps.

[0066] According to a modeling approach, γ-cyclodextrin offers a better trade-off between its solubility in water and / or ethanol, on the one hand, and its cavity size, on the other hand, compared to the other cyclodextrins mentioned. In addition, particularly good enzymatic degradation was observed, which is explained in the modeling approach by the fact that the γ-cyclodextrin scaffold has a certain flexibility due to its size. Therefore, according to the lock-and-key principle, this molecule can be particularly well stored in the active pocket of the enzyme.

[0067] A mixture consisting of 10 wt% substituted or unsubstituted β-cyclodextrin and 90 wt% substituted or unsubstituted γ-cyclodextrin is particularly preferred, especially for the separation of cyclodextrin-AOS complexes from the liquid phase resulting from the starting mixture. This will be discussed in more detail below. When nonvolatile apolar organic substances, especially bitter substances, are to be obtained by the extraction of the present invention with cyclodextrin, it is preferable to use high proportions of up to exclusively γ-cyclodextrin. Thus, the present invention also offers the possibility of selectively removing nonvolatile aromatic substances, such as bitter substances, from extracts containing organic substances with at least one apolar group under conditions unfavorable to their complex formation, in particular by using an ethanol content of less than 40 vol% and / or exclusively γ-cyclodextrin.

[0068] By using a mixture of different cyclodextrins, such as a mixture of β-cyclodextrin and γ-cyclodextrin, the present invention allows bitter substances to be complexed to a greater extent, or almost exclusively, with γ-cyclodextrin molecules, and other aromatic substances to a greater extent, or almost exclusively, with other cyclodextrin molecules. Figuratively speaking, the "guest" seeks the optimal "host" for complexation. Thus, the ratio of the amounts of various cyclodextrins can affect the composition of the organic substance containing at least one apolar group that is removed.

[0069] Cyclodextrin can be used not only in the form of a mixture with water, but also in at least one of several other solvents, such as glycerol, propylene glycol, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), ethylene glycol, etc. Solid cyclodextrin is preferred so that the cyclodextrin can be easily handled and added to the starting mixture as a solid. Furthermore, using cyclodextrin as a solid does not cause additional dilution of the starting mixture by the solvent introduced with the cyclodextrin.

[0070] The amount of cyclodextrin can vary within a fairly wide range, but is preferably from about 1 wt % to about 100 wt %, more preferably from about 3 wt % to about 50 wt %, and most preferably from about 3 wt % to about 20 wt %, based on the amount of the starting mixture.

[0071] The mixing of the at least one cyclodextrin with the starting mixture can be accomplished using any device known to those skilled in the art, such as a mechanical stirrer, a mechanical disperser, or an ultrasonic disintegrator. Those skilled in the art will select the temperature and holding time parameters in coordination with the efficiency of complex formation and taking into account the economics of the process.

[0072] Generally, the temperature and holding time can vary widely. Preferred temperatures range from about 4° C. to about 25° C., preferably from about 4° C. to about 15° C., more preferably from about 4° C. to about 10° C., and most preferably at a temperature of about 6° C. Holding times may range from 20 minutes to 72 hours, and most preferably up to 48 hours.

[0073] In the context of the present invention, it has been found that cyclodextrins in aqueous and alcoholic media are capable of reversibly accommodating organic substances containing at least one non-polar group in their cavities, selectively complexing approximately 60% to 99% by weight of the aromatic substances present in the starting mixture, while the majority of the remaining components of the starting mixture remain in the liquid phase. This discovery of reversible accommodation of organic substances containing at least one non-polar group is utilized in the method according to the present invention for the efficient selective separation of these substances.

[0074] Process (c) In step (c), the cyclodextrin-AOS complex formed in step (b) is separated from the liquid phase. The cyclodextrin-AOS complex formed is very stable, meaning that nonpolar organic substances bound to the complex, such as aromatic substances, have little tendency to re-leave the complex or the cyclodextrin cavity. Therefore, the complex is stable enough to be concentrated in an aqueous medium by filtration techniques. In principle, the complex can be separated using any of the processes and techniques commonly used for solid-liquid separation without releasing the guest in the process. Preferred separation techniques for the separation of the cyclodextrin-AOS complex in step (c) are filtration and / or sedimentation and / or centrifugation, with filtration such as vacuum filtration or nanofiltration or ultrafiltration being particularly preferred. Reverse osmosis is another separation process suitable for the separation of the cyclodextrin-AOS complex in step (c).

[0075] In a further advantageous embodiment, the method according to the invention comprises the steps of: (c1) a further step of allowing the cyclodextrin-AOS complex to settle before separating it from the liquid phase; wherein the resting step according to step (c1) is carried out in particular for a period of at most 24 hours, preferably for a period of at most 12 hours, most preferably for a period of 2 hours.

[0076] Settling the cyclodextrinization starting mixture in which the cyclodextrin-AOS complex has formed or has formed promotes separation of the complex. Surprisingly, it has been found that settling achieves faster sedimentation and better filtration behavior. Settling causes sedimentation, which makes it possible to remove a clear phase essentially free of the cyclodextrin-AOS complex from above the sediment containing the cyclodextrin-AOS complex. This reduces the total volume that needs to be separated, for example, filtered, in step (c). Step (c) can also be performed by settling alone, by removing the supernatant to obtain the phase containing the cyclodextrin-AOS complex.

[0077] In solid-liquid separation processes, filter aids are usually used to improve filtration. They can be incorporated into filter plates, filter layers, or filter cartridges, or can be added directly to the suspension to be filtered as sedimentation aids (diatomaceous earth, cellulose, bentonite, or other cationic flocculants). Physically, for example, centrifuges can be used to accelerate sedimentation and thus separation. Other processes that use filter aids include precoat filtration. The most important examples here are diatomaceous earth filtration or perlite filtration. The filter layer is pre-washed on a horizontal or vertical support layer. The actual filtration is carried out with the constant addition of filter aid to prevent the filter from clogging.

[0078] The filter aid ensures the permeability of the filter and prevents the filter surface from gradually becoming clogged with accumulated filter cake, especially in filters consisting of filtration layers. The filtrate is usually the desired product, and the retentate or filter cake is discarded.

[0079] In the context of the present invention, it is possible to dispense with the use of the conventional filter aids mentioned above, since the cyclodextrin-AOS complex is isolated as the filter cake, which is the desired "product" of separation step (c). If the conventional filter aids mentioned above were used, they would form a retentate together with the cyclodextrin complex. The filter aids would have to be separated from the filter cake in a further process step. While this is possible within the scope of the present invention, avoiding the mixing of the filter aid with the final product would be a significant improvement from an economic, ecological, and nutritional point of view.

[0080] Studies on the sedimentation behavior and filtration properties of various cyclodextrins have shown that complexes with β-cyclodextrin settle faster than similar complexes with γ-cyclodextrin. Furthermore, complexes with β-cyclodextrin exhibit better filtration behavior than similar complexes with γ-cyclodextrin, as they aggregate with significantly less or no clogging of the filter surface.

[0081] However, the products obtained with β-cyclodextrin do not always have favorable organoleptic properties: perfumes obtained with γ-cyclodextrin, on the other hand, have better organoleptic qualities, but they settle slowly and clog the filter surfaces very quickly.

[0082] Therefore, mixtures of β-cyclodextrin with other cyclodextrins were investigated and it was surprisingly found that the use of even a relatively small amount of β-cyclodextrin, for example 10 wt% β-cyclodextrin based on the total mass of cyclodextrin used, can significantly accelerate sedimentation. By adding β-cyclodextrin to α-cyclodextrin and / or γ-cyclodextrin and / or δ-cyclodextrin, preferably γ-cyclodextrin, the present invention provides a method for accelerating sedimentation and centrifugation, respectively.

[0083] According to one advantageous embodiment, it is contemplated that the proportion of β-cyclodextrin in the total mass of cyclodextrins is in the range of 0.5 wt% to a maximum of 60 wt%, preferably in the range of 2 wt% to 50 wt%, most preferably in the range of 5 wt% to 15 wt%. Thus, within the scope of the present invention, the composition of the cyclodextrin can be selected for each application with regard to the specific organic substance(s) containing at least one apolar group obtained from the starting mixture and / or the speed of the separation process.

[0084] For example, in combination with 90 wt% γ-cyclodextrin, sedimentation was accelerated from 1 day to 2 hours. The aromas obtained using a mixture of 10 wt% β-cyclodextrin and 90 wt% γ-cyclodextrin had the same sensory qualities as the corresponding aromas extracted from pure γ-cyclodextrin.

[0085] As an example, sedimentation velocity was measured using an ethanolic beer extract system containing 60% ethanol by volume and with aroma concentrations ranging from 5% to 8% by weight. In this case, all aroma-active components detected in the GC spectrum were collectively referred to as "aroma." The suspension obtained after complexation, containing the cyclodextrin-aroma complexes in the ethanolic extract, was simultaneously transferred to the same measuring cylinder. The volume of the sediment was measured by reading the graduations on the measuring cylinder at 30-minute intervals. In this way, the sedimentation behavior of the cyclodextrin-complexed substances according to the present invention can be determined.

[0086] Separation of the cyclodextrin-AOS complex from the liquid phase resulting from the starting mixture can be an important step in increasing the yield of organic substances containing at least one nonpolar group. For example, when obtaining aroma substances from an ethanol-containing starting mixture, the efficiency of subsequent treatment of the complex, especially enzymatically, can be influenced by solid-liquid separation of the cyclodextrin-aroma substance complex from the ethanol medium.

[0087] Solid-liquid separation forms a filter cake containing cyclodextrin-AOS complex.When separating aromatic substances from ethanol medium, if this filter cake is not "dry" enough, the alcohol still present may hinder the subsequent enzymatic decomposition of cyclodextrin complex.In addition, in the case of single-layer filtration, the filter cake gradually accumulates, which may clog the filter layer, making the separation process uneconomical due to time constraints.In this situation, filter aids can prevent the accumulated filter cake from clogging the filter layer.

[0088] Surprisingly, the inventors have found that β-cyclodextrin can function not only as an extractant for non-polar organic compounds complexed as guests, but also as a filter aid. β-cyclodextrin as a filter aid advantageously remains in the filter cake and can be further processed by enzymes to be broken down or degraded into substances harmless for food use. Therefore, β-cyclodextrin as a filter aid is similar to the substances used for complexing non-polar organic compounds, and has the same chemical and physical properties.

[0089] The inventors were surprised to discover that β-cyclodextrin, like γ-cyclodextrin, can bind aroma substances to itself and also function as a filter aid. After β-cyclodextrin has been used as an extractant and a filter aid in the method of the present invention, it can be degraded using enzymes in the context of the present invention, simultaneously re-releasing the complexed aroma substances. The only degradation products of cyclodextrin are mono-, di-, and oligosaccharides, which are commonly found in foods or luxury goods and may remain there. This will be discussed in more detail below. Thus, the problems of filter aid contamination and residue in such products are avoided by the present invention.

[0090] Thus, the product has the same nutritional purity and quality as when γ-cyclodextrin alone is used, while at the same time, processability is significantly improved. Here, quality specifically refers to the sensory quality of non-polar organic substances. For example, in the case of aromatic substances, an excessively high content of β-cyclodextrin can result in a change in flavor profile due to the selective complexation of some flavor components at the expense of others. However, the sweetness caused by the degradation products of cyclodextrin is usually negligible, given the final dosage.

[0091] The present invention was tested on the above-mentioned ethanolic beer extract. The test included complexation of aroma substances with a γ / β-cyclodextrin mixture, measurement of the sedimentation rate, drying of the filter cake, enzymatic degradation of the cyclodextrin to release the aroma substances again, analytical determination of the aroma concentration and aroma profile, and sensory evaluation of the samples thus produced. The aroma concentration was quantified using gas chromatography as the sum of all aroma substances typical of beer. The amount of residual cyclodextrin was measured by HPLC (high-performance liquid chromatography) using an RI detector. Thus, even a mere 10 wt% β-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 aroma extract of aroma substances typical of beer, concentrated compared to the starting mixture. It was successfully applied at a dosage of 1:1000 to beer with an alcohol content of 0.0% by volume. The term "successfully" means that the addition of the final product results in an aroma profile characteristic of beer. This dosage is low enough to result in an ethanol content of less than 0.05 vol% in the final application, allowing it to be declared a "0.0 vol%" beer, and the final product is clear, making it easy to use in typical clear beers such as Pils.

[0092] The present invention therefore also relates to the use of β-cyclodextrin as a filter aid.

[0093] The separated liquid phase as obtained by separation of the cyclodextrin-AOS complex in step (c) may optionally be recombined with at least one cyclodextrin, and in particular recycled to step (a), in order to maximize the yield.

[0094] According to a further embodiment of the invention, it is contemplated that the cyclodextrin-AOS complexes separated in step (c) are diluted with water in step (d1) to reach the desired final concentration of one or more fragrance substances or cyclodextrin-AOS complexes, prior to the treatment, in particular with enzymes, in step (d) described in more detail below. In this way, conditions can be improved to allow the most complete release of the extracted non-polar organic substance(s).

[0095] Process (d) In step (d), the solid phase separated in step (c), containing the cyclodextrin-AOS complexes, is subjected to an enzymatic treatment with at least one enzyme selected from the group comprising an enzyme having amylase activity, preferably an α-amylase, most preferably a fungal α-amylase, a debranching enzyme, in particular a pullulanase and / or an isoamylase, and a mixture of at least two of these enzymes, in order to hydrolyze the cyclodextrins in these complexes.

[0096] The resulting mixture is optionally filtered.

[0097] During targeted enzymatic treatment of the host-guest complex, the ring structure of the cyclodextrin is broken, and the previously complexed and optionally enriched target substance is released again without thermal energy. The available α-, β-, γ-, and δ-cyclodextrins, as well as the complexes formed with these cyclodextrins as hosts, can be partially and selectively hydrolyzed by different enzymes, in particular different amylases. As a result, the present invention allows the design of processes that allow the cascade release of non-polar organic substances, such as aroma substances.

[0098] After step (d), a composition loaded with at least one aromatic substance is obtained, which also contains degradation products of cyclodextrin, such as mono-, di- and oligosaccharides, which are commonly found in food products and may remain therein or may be removed as needed.

[0099] This treatment, in particular the enzymatic treatment, according to step (d) of the method according to the invention is carried out in the presence of water. In one embodiment of the invention, step (d) is preceded by a step (d1) in which the solid phase separated in step (c) is mixed with water until the desired final concentration of one or more aromatic substances is reached. The final concentration preferably used is at least 100 wt. % and up to about 3750 wt. % water, based on the total weight of the solid phase separated in step (c).

[0100] In one embodiment of the present invention, the water added in step (d1) has a temperature in the range of 4° C. to 80° C., preferably in the range of 20° C. to 60° C. The pH value can be adjusted depending on the enzyme used, and is preferably in the range of pH 3.5 to pH 7.5, most preferably 4.5, or 5.2 to pH 5.6.

[0101] The resulting aqueous mixture is then treated with at least one enzyme selected from the group comprising an enzyme having amylase activity, preferably an α-amylase, most preferably a fungal α-amylase, a debranching enzyme, in particular a pullulanase and / or an isoamylase, and a mixture of at least two of the mentioned enzymes, preferably a mixture of two amylases, and an enzyme capable of degrading cyclodextrins as a side activity.

[0102] By using different enzymes, in particular different amylases, it is possible to selectively hydrolyze the α-, β- and / or γ-cyclodextrins used in the complex, thereby enabling a cascade-like release process of the aroma substances.

[0103] Alpha-amylases are particularly preferred, and most preferred for the treatment are fungal alpha-amylases, which are preferably used alone.

[0104] Fungal α-amylases for use in accordance with the present invention are derived from microorganisms such as Aspergillus niger and Aspergillus oryzae. Suitable commercial products of fungal α-amylases are sold by Novozymes under the name "Fungamyl™". Another suitable amylase is sold by Novozymes under the name "Dextrozyme GA".

[0105] It is known that enzymes from different sources exhibit different reactivities. The amount of enzyme required for each mixture varies depending on the mixture and the enzyme. The preferred amount of enzyme used for hydrolysis of cyclodextrin in the cyclodextrin-fragrance complex essentially depends on the content of the solid phase separated in step (c) containing the cyclodextrin-fragrance complex, and may also depend on the activity of the enzyme.

[0106] In the context of the present invention, it has been found that a preferred amount of the at least one enzyme is between 5 and 1000 FAU per gram of solid phase separated in step (c). The unit "FAU" stands for "Fungal α-Amylase Unit" and is a measure of the activity of α-amylase, as used by Novozymes, for example, in the enzyme Fungamyl™. More precisely, under standard conditions (substrate: soluble starch, incubation time 7 to 20 minutes, temperature 37°C, pH 4.7), 1 FAU of the enzyme degrades 5.26 g of starch per hour.

[0107] Treatment conditions such as temperature and time can be varied over a wide range, particularly in step (d), but temperatures of about 4°C to about 80°C, preferably about 20°C to about 60°C, have been found to be advantageous, and treatment times generally range from 0.5 to 50 hours. The pH value can be between 3.5 and 7.5, most preferably between 5.2 and 5.6. Incubation in a closed container is particularly preferred.

[0108] After enzyme treatment, the maximum final cyclodextrin concentration is about 0.1 wt %, based on the total weight of the aqueous mixture. The amount of cyclodextrin in the mixture is determined using known methods, i.e., by HPLC.

[0109] Additionally, additional enzymes can be used, for example, suitable debranching enzymes such as pullulanase and / or isoamylase can be used prior to treatment with the at least one amylase and / or in combination with the at least one amylase.

[0110] In a further embodiment of the invention, the enzyme is inactivated and / or separated from the composition containing the non-polar organic material released from the cyclodextrin-AOS complex by the enzymatic treatment. For example, propylene glycol (PG) or glycerol can be used for this purpose.

[0111] The subject of the present invention is at least one organic substance comprising at least one apolar group selected from the group comprising volatile aromatic substances, non-volatile aromatic substances, such as bitter substances, oil fractions, fat fractions and / or wax fractions, colorants and adhesives, in particular those from plant and / or animal starting materials, as well as mixtures of at least two of the substances mentioned; at least one saccharide having a chain length of 6 or 7 or 8 or 9 glucose units, and optionally at least one degradation product of cyclodextrin, in particular glucose and / or maltose, The present invention further includes compositions, particularly those produced by the methods as described above, comprising:

[0112] Within the scope of the present invention, in an advantageous simple embodiment, the composition may consist of at least one organic substance containing at least one apolar group, at least one saccharide with a chain length of 6 or 7 or 8 or 9 glucose units, and optionally at least one degradation product of cyclodextrin, in particular glucose and / or maltose.

[0113] In a preferred embodiment of the invention, the composition has an ethanol content of 0.0 vol%.

[0114] The composition is preferably an aqueous composition. In this case, the composition according to the present invention can be further processed depending on the intended use. For example, it is possible to inactivate at least one enzyme contained in the composition by heating and / or changing the pH value, or by using PG or glycerol. Depending on the at least one non-polar organic substance and the type of enzyme or organism used to decompose cyclodextrin, the non-polar organic substance itself can also cause or support inactivation.

[0115] At least one cyclodextrin and enzyme degradation product can be removed from the composition of the present invention by a common "downstream" process, such as liquid / liquid extraction, to obtain undiluted non-polar organic substances, so that the resulting composition can contain only at least one non-polar organic substance, particularly at least one aromatic substance, which can be of animal or plant origin.When the solubility of the non-polar organic substance in water is exceeded, this substance will be separated as an oil phase or solid.In this way, the non-polar organic substance can be further separated from the aqueous sugar-containing phase.

[0116] Another possibility for separating at least one degradation product of the cyclodextrin is their oxidation to an acid, which can then be removed by the formation of an adsorbent, an anion exchanger or a chelate.

[0117] The present invention therefore also makes it possible to use the composition according to the invention, e.g. loaded with aroma substances, for introducing at least one aroma substance and / or one organic substance comprising at least one apolar group, in particular selected from the group comprising volatile aroma substances, non-volatile aroma substances, e.g. bitter substances, oil fractions, fat fractions and / or wax fractions, colorants and adhesives, in particular those from plant and / or animal starting materials, as well as mixtures of at least two of the mentioned substances, into a food, luxury product, cosmetic or pharmaceutical product, preferably a food or beverage, most preferably a food or luxury product or beverage or cosmetic or pharmaceutical product having an ethanol content of 0.0 vol.%.

[0118] Thus, the present invention also provides a method for flavoring and / or stabilizing a product, be it a food, luxury item, cosmetic, or pharmaceutical, in which the above-described composition or a composition produced by the above-described method is added to the product to be flavored. In this case, such a product can be advantageously advertised with the label "alcohol-free" (i.e., alcohol content 0.0 vol%) and / or "FTNS." Depending on the resulting organic substance containing at least one nonpolar group, the present invention provides not only a method for flavoring, but also a method for stabilizing a product using, for example, an oil fraction, a fat fraction, and / or a wax fraction as a coating or coating ingredient, and / or a phenolic substance, for example, for color stabilization.

[0119] In the method according to the invention for flavoring and / or stabilizing a product, in particular a food, luxury item, cosmetic or pharmaceutical product, the composition according to the invention, in particular produced by the method described above, is brought into contact with the product to be flavored and / or stabilized, this "contacting" being achieved by any process known to the skilled person suitable for this purpose, preferably by mixing.

[0120] A further subject of the present invention is a food or luxury product or cosmetic or pharmaceutical product containing the composition according to the invention, preferably produced by the above-described method. Beverages, most preferably beer, and food or luxury products or cosmetic or pharmaceutical products that can be advertised as "alcohol-free" (i.e., alcohol content 0.0 vol%) and / or "FTNS" are particularly preferred.

[0121] The present invention will be described in more detail with reference to the accompanying drawings and exemplary embodiments, without being limited to each of the specific embodiments described. The present invention also relates to any combination of preferred embodiments, unless they are mutually exclusive. The term "about" or "approximately" when used in conjunction with a numerical value is intended to include a value that is at least 10% higher or lower, or 5% higher or lower, and in each case 1% higher or lower. [Brief explanation of the drawings]

[0122] [Figure 1] 1 is a schematic diagram of a first embodiment of the method according to the invention using the example of separating aromatic substances from an aqueous solution, for example a fermentation broth. [Figure 2] FIG. 1 is a schematic diagram of a second embodiment of the method according to the invention using the example of separating aromatic substances from an ethanol extract obtained, for example, from SPE. [Figure 3] Photographs of comparative sedimentation rates of a gamma-cyclodextrin-fragrance complex (left), a beta-cyclodextrin-fragrance complex (center), and the corresponding complex in a mixture of 10 wt% beta-cyclodextrin and 90 wt% gamma-cyclodextrin, based on the total mass of cyclodextrin (right). DETAILED DESCRIPTION OF THE INVENTION

[0123] FIG. 1 shows a schematic diagram of an embodiment of the method according to the present invention for selectively separating at least one organic substance (AOS) containing at least one apolar group. First, in step (a), a starting mixture 10 containing apolar organic substances in the form of an aromatic substance 1 in an aqueous solution is prepared. Such a water-based aromatic substance 3 can be a fermentation broth, e.g., a broth with a particularly low aromatic substance concentration and optionally containing at least one solvent. In step (b), the starting mixture 10 consisting of the aromatic substance 1 in water 3 is contacted with at least one cyclodextrin 2, which was added as a solvent in step (a). As a result of contacting the at least one cyclodextrin 2 with the at least one apolar organic substance 1 of the starting mixture 10, at least one cyclodextrin-AOS complex 12 is obtained. Due to the formation of the complex, the apolar organic substance in the form of the aromatic substance is extracted from the solvent water 3 of the starting mixture 10. The complex 12 is present in the aqueous phase.

[0124] 1 also shows an optional continuation of the method according to the invention. In step (c), the cyclodextrin-AOS complex 12 is separated from the liquid phase by removing water 3. Subsequently, in step (d), the fragrance 1 is released from the cyclodextrin-AOS complex 12 by enzymatic treatment. A composition 6 loaded with apolar organic substances, here fragrance 1, is obtained, which comprises concentrated fragrance 1 and saccharides 20 as by-products of the enzymatic treatment.

[0125] 2 shows a schematic diagram of a further embodiment of a method according to the present invention for selectively separating at least one non-polar organic substance (AOS). First, in step (a), a starting mixture 10 is prepared, e.g., containing an ethanol extract from solid-phase extraction (SPE) in the form of an aroma substance 1 in an ethanol-containing solution 4, e.g., containing 80 vol.% alcohol (ethanol). In step (b), the starting mixture 10, consisting of the aroma substance 1 in the ethanol solution 4, is contacted with at least one cyclodextrin 2. As a result of the contact of the at least one cyclodextrin 2 with the at least one non-polar organic substance 1 of the starting mixture 10, at least one cyclodextrin-AOS complex 12 is obtained. Due to the formation of the complex, the non-polar organic substance in the form of the aroma substance is extracted from the ethanol-containing solution 4 of the starting mixture 10. The complex 12 is present in the aqueous alcohol-containing phase 4.

[0126] 2 also shows an optional continuation of the method according to the invention. In step (c), the cyclodextrin-AOS complex 12 is separated from the liquid phase by removing ethanol 40. Subsequently, in step (d), the odorant 1 is released from the cyclodextrin-AOS complex 12 by enzymatic treatment. For this purpose, in step (c), water 3 is added to the solid phase consisting of the cyclodextrin-in-aromatic complex 12 to create conditions suitable for the enzymatic treatment. A composition 6 loaded with a non-polar organic substance, here odorant 1, is obtained from step (d). It comprises concentrated odorant 1 and saccharides 20 as by-products of the enzymatic treatment. Depending on the process control and the ethanol concentration that can be tolerated in the composition for the respective application, the composition can contain, for example, up to 10 vol.% ethanol.

[0127] Figure 3 shows three photographs, from left to right, depicting a test setup consisting of three cylinders. The cylinders were filled with suspensions of cyclodextrin-AOS complexes. Each cylinder on the left contained a complex of γ-cyclodextrin, each cylinder in the middle contained a complex of β-cyclodextrin, and each cylinder on the right contained a complex in a mixture consisting of 10 wt% β-cyclodextrin and 90 wt% γ-cyclodextrin, based on the total mass of cyclodextrin (right). In addition to these cyclodextrins, the suspensions shown contained an ethanol-containing beer extract. Each measuring cylinder contained 150 g of beer extract and 9 g of β- or γ-cyclodextrin or a 9 g mixture of β-cyclodextrin and γ-cyclodextrin. The suspensions were stirred for 48 hours at 6 °C in the presence of different cyclodextrins or cyclodextrin mixtures and then simultaneously transferred to measuring cylinders.

[0128] After the start of the comparative experiment at time 0 h (left photo), the settling of particles from the suspension can be seen over a period of 0.5 h (middle photo) to 1 h (right photo), showing that particles from the suspension containing β-cyclodextrin already settle after 0.5 h, and therefore twice as fast as the suspension containing γ-cyclodextrin, where a clear supernatant visually distinguishable from the sediment at the bottom of the cylinder only appears after the 1 h test period.

[0129] Surprisingly, suspensions containing a mixture of only 10 wt. % β-cyclodextrin and 90 wt. % γ-cyclodextrin, based on the total mass of cyclodextrins, i.e., containing primarily γ-cyclodextrin, exhibit the same visual impression of sedimentation behavior as suspensions containing only β-cyclodextrin as the cyclodextrin. Thus, according to the present invention, the addition of β-cyclodextrin to γ-cyclodextrin in a ratio of less than 1:1 is sufficient to increase the sedimentation rate of a suspension containing γ-cyclodextrin to a value comparable to the sedimentation rate of a suspension containing only β-cyclodextrin. [Example]

[0130] Exemplary embodiments: For example, Example 1 for flavoring 0.0 vol% alcohol beer An ethanol-containing (70%-80% by volume) solid extract (1 kg, 4 g / L of fragrance) containing isoamyl alcohol, isoamyl acetate, phenylethyl alcohol, hexanoic acid, ethyl hexanoate, and other fragrances is mixed with 60 g of α-, β-, and / or γ-cyclodextrin, preferably γ-cyclodextrin. The mixture is stirred at 6°C for 48 hours. The fragrance-cyclodextrin complex is separated by filtration and dried. To maximize yield, the ethanol solid extract may be mixed again with α-, β-, and / or γ-cyclodextrin, preferably γ-cyclodextrin, at 30 g per kg of solid extract. The resulting alcohol-free filter cake is then taken up in water and treated with enzymes to release the fragrance. For this purpose, each ml of the aqueous mixture is mixed with 1 μl of amylase (Fungamyl™ from Novozymes) (1 μl of amylase per ml of aqueous mixture) and incubated in a closed vessel at 55° C. and pH 5.2 for 48 hours (maximum final cyclodextrin concentration is 0.1 wt %, based on the total weight of the aqueous extract). Due to the low solubility of the extracted aroma compounds in water, the resulting two-phase mixture may be homogenized with propylene glycol (up to 1:1 w / w) if necessary, followed by filtration.

[0131] The aroma-rich alcohol-free product (5 g / l) thus obtained can now be used to flavor beer, for example 0.0 vol.% alcohol beer (application amount: 0.2:1000).

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

[0133] The concentration of non-polar organic substances, e.g. the concentration of aromatic substances, of the compositions according to the invention, in particular of the alcohol-free extracts, is determined by GC-FID (2 g of sample is extracted with 2 g of cyclohexane. The organic phase is dried over Na2SO4, mixed with an internal standard and analyzed).

[0134] Example 2 for flavoring 0.0 vol% alcohol beer The fermentation broth containing isoamyl acetate or 4-vinylguaiacol is mixed with β-cyclodextrin (α- and / or γ-) in a molar equivalent amount relative to the aroma substance, and the mixture is stirred at 6°C for 48 hours to obtain a stable aroma-cyclodextrin complex.

[0135] The fragrance-cyclodextrin complex is separated by filtration or concentrated in the retentate.

[0136] The release of aroma substances is achieved by enzymatic treatment of the aroma-cyclodextrin complex. For this purpose, the aqueous mixture is mixed with 1 μl of amylase (Novozymes' Fungamyl™) per ml (1 μl amylase per ml of aqueous mixture) and incubated in a sealed container at 55° C. and pH 5.2 for 48 hours (maximum final cyclodextrin concentration is 0.1 wt %, based on the total weight of the aqueous extract). Due to the low solubility of the extracted aroma compounds in water, the resulting two-phase mixture can be homogenized with propylene glycol (up to 1:1 w / w) if necessary, followed by filtration.

[0137] The resulting aroma-rich, alcohol-free product can now be used to flavor beer, e.g., 0.0 vol% alcohol beer. The final cyclodextrin concentration is determined by HPLC (detector: RI; separation column: Phenomenex Polysep GFC-P 2000; eluent: water (isocratic); flow rate: 0.8 ml / min; pressure: 20 bar; oven temperature: 55 °C; run time: 30 min). The aroma concentration of the alcohol-free extract is determined using GC-FID (2 g sample was extracted with 2 g cyclohexane. The organic phase was dried with Na2SO4, mixed with an internal standard, and analyzed).

[0138] Example 3 for flavoring a 0.0 vol% alcoholic beverage A solid extract containing ethanol (70-80 vol%) (or spirits or liqueurs (15-96 vol%)) containing isoamyl alcohol, isoamyl acetate, phenylethyl alcohol, and other aromatic substances was mixed with 60 g / L of a mixture of β-cyclodextrin and γ-cyclodextrin (1:9). The mixture was stirred overnight at 6°C to obtain a sufficiently stable host-guest complex. The mixture was then left to settle for 2 hours (without β-cyclodextrin, this process can take up to 1 day).

[0139] After settling, the clear phase was removed and the precipitate was dried by filtration.

[0140] To maximize the yield, the clear ethanolic solid extract, depleted of aroma substances, may be mixed again with a mixture of β-cyclodextrin and γ-cyclodextrin (1:9), allowed to settle, and the clear phase removed after sedimentation, followed by filtration.

[0141] The alcohol-free filter cake thus obtained was taken up in water and treated with an enzyme to release the aroma substances. For this purpose, 1 μl of amylase Dextrozyme GA (Novozymes) was added per ml of the aqueous mixture and incubated in a sealed container at 55° C. and pH 4.5 for 48 hours. Finally, the resulting mixture was filtered.

[0142] The aroma-rich alcohol-free product so obtained can now be used for flavoring purposes, for example in small amounts in the range of 0.01:1000 to 50:1000, for example to flavor 0.0 vol% alcohol beer.

[0143] Example 4 for Extraction of Apple Wax-Containing Fraction from Dried Apple Pomace 1600 g of dried apple pomace was mixed with 8000 g of a 36 wt% aqueous cyclodextrin solution (β-cyclodextrin and γ-cyclodextrin in a 1:1 ratio, i.e., 18 g of β-cyclodextrin and 18 g of γ-cyclodextrin per 1 L of water) and kneaded at room temperature for 2 days. The wet apple pomace was then filtered, and the filtrate, enriched in cyclodextrin-apple wax complex, was collected as a suspension.

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

[0145] The extract (182.65 g) thus obtained was mixed with 200 g of water and incubated with the amylase "Dextrozyme GA" (400 μl) at pH 4.5 and 55° C. for 48 hours. The mixture was then filtered using a vacuum and dried to constant weight in a rotary evaporator. The apple wax-rich fraction thus obtained (16 g) was successfully tested as a component of a peeling agent for fruit gums. This means that the apple wax-rich fraction thus obtained can be used as a peeling agent in the same way as peeling agents containing wax (also apple wax) obtained by other previously known methods.

[0146] Example 5: Extraction of an Aroma-Containing Oil Fraction from Hop Cones 50 g of dried hop cones of the "Hercules" variety were mixed with 750 g of an 18 wt % aqueous solution of β-cyclodextrin and kneaded at room temperature for 2 days. The wet hop cones were then filtered, and the filtrate, enriched in cyclodextrin-oil complexes, was collected as a suspension.

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

[0148] The extract thus obtained (28.99 g) was mixed with 15 g of water and incubated with the amylase "Dextrozyme GA" (88 μl) at pH 4.5 and 55° C. for 48 hours. The aroma-rich extract thus obtained has a high aroma load and can be used, for example, in an amount of 1:1000, to flavor alcohol-free beer.

[0149] Example 6 for Extraction of Aromatic Substances from Gentian Root Extract Containing Ethanol A gentian root extract with an alcohol content of 60 vol% was mixed with 6 wt% γ-cyclodextrin in solid form. The mixture was stored at 6°C overnight. In this way, a sufficiently stable host-guest complex was obtained.

[0150] In step (c) of the method according to the present invention, the host-guest complex was separated by filtration. Water was added to the filter cake. The release of the aroma substances was achieved by enzymatic treatment of the host-guest complex. For this purpose, the aqueous mixture was mixed with 1 μl of amylase "Dextrozyme GA" (Novozymes) per ml and incubated in a sealed container at 55° C. for 48 hours. Finally, the resulting mixture was filtered.

[0151] The resulting aroma-rich product can now be used in small amounts for flavoring purposes in products having an alcohol content of, for example, 0.0 vol.% ethanol.

[0152] It will be clear to those skilled in the art that the invention is not limited to the embodiments described above, but rather can be varied in many ways, and in particular the features of the embodiments described individually can also be combined with one another or substituted for one another. [Explanation of symbols]

[0153] 1 Organic substances containing at least one non-polar group, abbreviated as "non-polar organic substances (AOS)"; aromatic substances, bitter substances, oils, fats, waxes, adhesives, colorants 10 Starting mixture 2. Cyclodextrin 12 Guest-host complexes of cyclodextrins and organic substances containing at least one nonpolar group (AOS), cyclodextrin-AOS complexes 20 saccharides 3 water 4. Ethanol-containing solutions 40 Ethanol 5. Enzymes 6 Composition

Claims

1. A method for flavoring and / or stabilizing a product that constitutes a food, luxury item, cosmetic, or pharmaceutical product, comprising: adding to the product to be flavored and / or stabilized at least one organic substance (AOS) (1) containing at least one apolar group and / or at least one or more aromatic substances, a composition (6) prepared by a method for selectively separating said composition (6); The method for selectively separating at least one organic substance (AOS) (1) containing at least one non-polar group and / or at least one or more aromatic substances comprises: (a) providing a starting mixture (10) containing at least one organic substance (AOS) containing at least one apolar group and / or at least one or more aromatic substances (1), and optionally at least one solvent (3; 4; 40); (b) contacting the starting mixture (10) with 3 wt % to 20 wt % of at least one cyclodextrin (2) relative to the amount of the starting mixture (10) at a temperature ranging from 4° C. to 15° C.; wherein at least one solvent is added during and / or after step (a) and / or step (b), As a result of contacting the at least one cyclodextrin (2) with the at least one organic substance containing 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 phase (3; 4), (c) a further step of separating the cyclodextrin-AOS complex and / or the cyclodextrin-fragrant substance complex (12) from the liquid phase; (d) the further step of treating the separated cyclodextrin-AOS complex and / or the cyclodextrin-fragrance complex (12) with an enzyme to obtain at least one organic substance (AOS) (1) containing at least one non-polar group and / or at least one fragrance-loaded composition (6); and performing an enzymatic treatment using at least one enzyme (5) selected from the group consisting of enzymes having amylase activity, α-amylase, fungal α-amylase, debranching enzyme, pullulanase and / or isoamylase, and a mixture of at least two of these enzymes; method.

2. 1. Use of a composition (6) produced by a process comprising selectively isolating at least one organic substance (AOS) (1) containing at least one apolar group and / or at least one or more aromatic substances, for introducing at least one aromatic substance and / or at least one organic substance (1) containing at least one apolar group into a food, beverage, luxury product, cosmetic or pharmaceutical product, the composition (6) being selected from the group consisting of volatile aromatic substances, non-volatile aromatic substances, bitter substances, oil fractions, fat fractions and / or wax fractions, colorants and adhesives, those from plant and / or animal starting materials, and mixtures of at least two of the aforementioned substances, The method for producing said composition (6) for selectively separating at least one organic substance (AOS) (1) containing at least one apolar group and / or at least one or more aromatic substances comprises the steps of: (a) providing a starting mixture (10) containing at least one organic substance (AOS) containing at least one apolar group and / or at least one or more aromatic substances (1), and optionally at least one solvent (3; 4; 40); (b) contacting the starting mixture (10) with 3 wt % to 20 wt % of at least one cyclodextrin (2) relative to the amount of the starting mixture (10) at a temperature ranging from 4° C. to 15° C.; wherein at least one solvent is added during and / or after step (a) and / or step (b), As a result of contacting the at least one cyclodextrin (2) with the at least one organic substance containing 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 phase (3; 4), (c) a further step of separating the cyclodextrin-AOS complex and / or the cyclodextrin-fragrant substance complex (12) from the liquid phase; (d) the further step of treating the separated cyclodextrin-AOS complex and / or the cyclodextrin-fragrance complex (12) with an enzyme to obtain at least one organic substance (AOS) (1) containing at least one non-polar group and / or at least one fragrance-loaded composition (6); and performing an enzymatic treatment using at least one enzyme (5) selected from the group consisting of enzymes having amylase activity, α-amylase, fungal α-amylase, debranching enzyme, pullulanase and / or isoamylase, and a mixture of at least two of these enzymes; use.

3. (b) contacting the starting mixture (10) with at least one cyclodextrin (2); wherein water is added in step (a) and / or step (b) and / or after step (b), 3. The method according to claim 1 or the use according to claim 2, wherein as a result of contacting 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 the liquid phase (3; 4).

4. 4. The method according to claim 1 or 3 or the use according to claim 2 or 3, 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, bitter substances, oil fractions, fat fractions and / or wax fractions, colorants and adhesives, as well as mixtures of at least two of the substances mentioned.

5. In the step (a) and / or the step (b), and / or after the step (b), adjusting the moisture content to a range of 15 vol% to 35 vol%, or 20 vol% to 30 vol%, and / or the ethanol content is adjusted to an ethanol content of at least 40 vol.% or in the range of 40 vol.% to 60 vol.%.

5. Use according to claim 2, 3 or 4.

6. (d1) a further step of diluting the cyclodextrin-AOS complex and / or the cyclodextrin-fragrant substance complex (12) separated in step (c) with water prior to treatment in step (d), i.e., enzymatic treatment; The method according to claim 1 or 3 to 5 or the use according to any one of claims 2 to 5, comprising:

7. 7. The method according to claim 1 or any one of claims 3 to 6 or the use according to any one of claims 2 to 6, characterized in that step (d) is carried out so that the concentration of cyclodextrin in the composition (6) loaded with at least one apolar organic substance (AOS) and / or at least one fragrant substance (1) is less than 0.5 wt. %, or less than 0.1 wt. %.

8. (e) a further step of filtering the mixture resulting from the treatment, in particular with an enzyme, of the cyclodextrin-AOS complex and / or the cyclodextrin-fragrant substance complex (12), and a composition (6) is obtained which is loaded with at least one organic substance (AOS) comprising at least one apolar group and / or at least one aromatic substance.

9. At least one solvent is used in step (a) and / or step (b) and / or after step (b), and the solvent is selected from the group consisting of water, C １ ~C ４ The method according to any one of claims 1 or 3 to 8 or the use according to any one of claims 2 to 7, wherein the solvent is selected from the group comprising alcohol, diethyl ether, acetone or mixtures thereof, or water, ethanol and mixtures thereof.

10. 10. The method according to claim 1 or 3 to 9 or the use according to claim 2 to 9, wherein the cyclodextrin (2) is 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 a mixture thereof, or a substituted or unsubstituted γ-cyclodextrin, or a mixture consisting of 10 wt % of substituted or unsubstituted β-cyclodextrin and 90 wt % of substituted or unsubstituted γ-cyclodextrin.

11. The method according to any one of claims 1 or 3 to 10 or the use according to any one of claims 2 to 9, wherein the separation of the cyclodextrin-AOS complex (12) in step (c) is carried out by centrifugation or filtration, or by vacuum filtration or nanofiltration.

12. (c1) a further step of allowing the cyclodextrin-AOS complex and / or the cyclodextrin-fragrant substance complex to settle before separating it from the liquid phase; The method according to any one of claims 1 or 3 to 11 or the use according to any one of claims 2 to 11, wherein the standing step according to step (c1) is carried out for a maximum of 24 hours.

13. 13. The method or use according to claim 12, wherein the resting step according to step (c1) is carried out for a maximum of 12 hours.

14. 13. The method or use according to claim 12, wherein the standing step according to step (c1) is carried out for a maximum of 2 hours.

15. 15. The method according to claim 1 or any one of claims 3 to 14 or the use according to any one of claims 2 to 14, wherein the separated liquid phase obtained by separating the cyclodextrin-AOS complex and / or the cyclodextrin-aromatic substance complex (12) in step (c) is mixed again with at least one cyclodextrin (2).

16. 13. The method or use according to claim 12, wherein the separated liquid phase obtained by separating the cyclodextrin-AOS complex and / or the cyclodextrin-aroma complex (12) in step (c) is recombined with at least one cyclodextrin (2) and recycled to step (a) to maximize the yield.

17. 17. The method according to any one of claims 1 or 3 to 16 or the use according to any one of claims 2 to 16, wherein the cyclodextrin-AOS complex and / or the cyclodextrin-fragrant substance complex (12) separated in step (c) is diluted with water in step (d1) to obtain a desired final concentration of one or more hydrophilic organic substances and / or one or more fragrance substances (1) before the enzymatic treatment in step (d).

18. 18. The method according to any one of claims 1 or 3 to 17 or the use according to any one of claims 2 to 17, wherein the at least one enzyme (5) is used in an amount ranging from 5 FAU to 1000 FAU per g of separation solid phase.

19. 19. The method according to any one of claims 1 or 3 to 18 or the use according to any one of claims 2 to 18, wherein β-cyclodextrin serves as the sole filter aid used in the method and not only as an extractant to which at least one organic substance comprising at least one apolar group (AOS) and / or at least one aromatic substance is complexed as guest.

20. 20. Use of a composition produced by the method of any one of claims 1, 3 to 19 for introducing at least one aromatic substance and / or at least one organic substance (1) comprising at least one non-polar group into a food or luxury product or beverage or cosmetic or pharmaceutical product having an ethanol content of 0.0 vol%.

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