METHOD FOR SELECTIVELY SEPARATING AT LEAST ONE ORGANIC SUBSTANCE COMPRISING AT LEAST ONE APOLAR GROUP, AND USE OF SAID SUBSTANCE IN A FOOD, LUXURY FOOD, COSMETIC, OR PHARMACEUTICAL PRODUCT

Abstract

A method for selectively separating at least one apolar organic substance, comprises: (a) providing a starting mixture (10) which contains at least one organic substance comprising at least one apolar group and optionally a solvent; (b) bringing the starting mixture into contact with at least one cyclodextrin to obtain a cyclodextrin-aromatic substance complex; and optionally (c) separating the complex from the liquid phase; and (d) treating the separated complex, in particular enzymatically, and optionally filtering the resulting mixture, whereby a composition loaded with at least one organic substance comprising at least one apolar group is obtained. The invention also relates to a composition loaded with at least one organic substance comprising at least one apolar group, and to the use of this composition in order to introduce at least one apolar organic substance into a food, luxury food, cosmetic product, or pharmaceutical product.

Claims

1.-22. (canceled)

23. 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, a composition (6) produced by a method for selectively separating at least one organic substance that comprises at least one apolar group (AOS) (1) and/or at least one or more aromatic substances; wherein the 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 comprises: (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); (b) bringing the starting mixture (10) into contact with at least one cyclodextrin (2); wherein at least one solvent 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 phase (3; 4); (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 consisting of enzymes with amylase activity, debranching enzymes, and mixtures thereof.

24. A composition, produced by a method for selectively separating at least one organic substance that comprises at least one apolar group (AOS) (1) and/or at least one or more aromatic substances; for introducing at least one aromatic substance and/or at least one organic substance (1) comprising at least one apolar group, selected from the group consisting of volatile aromatic substances, non-volatile aromatic substances, oil, fat and/or wax fractions, colorants, and adhesives, and mixtures thereof; into a food, luxury food, cosmetic product, or pharmaceutical product, using the 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 comprises: (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); (b) bringing the starting mixture (10) into contact with at least one cyclodextrin (2); wherein at least one solvent 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 phase (3; 4); (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 consisting of enzymes with amylase activity, debranching enzymes, and mixtures thereof.

25. The method of claim 23, wherein the at least one organic substance (1) comprising at least one apolar group is selected from the group consisting of volatile aromatic substances, non-volatile aromatic substances, oil, fat and/or wax fractions, colorants, adhesives, and mixtures thereof

26. The method of claim 23, wherein 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 % is adjusted; and/or an ethanol content is adjusted so as to be at least 40 vol %.

27. The method of claim 23, 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 treatment in step d).

28. The method of claim 23, wherein step (d) is performed such that a 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 %.

29. The method as claimed in claim 23, comprising a further step of (e) filtering the mixture resulting from the 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.

30. The method as claimed in claim 23, 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 consisting of water, C.sub.1-C.sub.4 alcohols, diethyl ether, acetone, and mixtures thereof.

31. The method as claimed in claim 23, wherein as a cyclodextrin (2), at least one substituted or unsubstituted -cyclodextrin, at least one substituted or unsubstituted -cyclodextrin, at least one substituted or unsubstituted -cyclodextrin, at least one substituted or unsubstituted -cyclodextrin, or mixtures thereof is used.

32. The method as claimed in claim 23, wherein the separating of the cyclodextrin-AOS complex (12) in step (c) is carried out by centrifugation or filtration.

33. The method as claimed in claim 23, 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 over a duration of up to 24 hours.

34. The method as claimed in claim 23, 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 is recirculated to step (a), to maximize yield.

35. The method as claimed in claim 23, wherein the cyclodextrin-AOS complex and/or the cyclodextrin-aromatic substance complex (12) separated in step (c) is prior to the 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).

36. The method as claimed in claim 23, wherein the enzymatic treatment in step (d) is performed using at least one enzyme (5) selected from the group consisting of alpha-amylase, pulluanase, isoamylase, and mixtures thereof.

37. The method as claimed in claim 23, 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.

38. A method, comprising: using the composition according to claim 24 for 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 %.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0141] FIG. 1 is a schematic diagram of a first embodiment of the method according to the invention, using the example of the separation of an aromatic substance from an aqueous solution, for example from a fermentate;

[0142] FIG. 2 is a schematic diagram of a second embodiment of the method according to the invention, using the example of the separation of an aromatic substance from an ethanolic extract such as, for example, resulting from an SPE; and

[0143] FIG. 3 shows photographs of a comparison test of the sedimentation rate of gamma-cyclodextrin-aromatic substance complex (left), beta-cyclodextrin-aromatic substance complex (middle), and corresponding complexes in a mixture consisting of 10 wt % beta- and 90 wt % gamma-cyclodextrin, based on the total mass of cyclodextrin (right).

DETAILED DESCRIPTION

[0144] FIG. 1 shows a schematic diagram of an embodiment of the method according to the invention for selectively separating at least one organic substance which comprises at least one apolar group (AOS). First, in a step (a), a starting mixture 10 is provided, which contains an apolar organic substance in the form of an aromatic substance 1 in aqueous solution. Such a water-based aromatic substance 3 may be a fermentate, for example, which may in particular have a low concentration of aromatic substance and optionally contains at least one solvent. In step (b), the starting mixture 10 consisting of aromatic substance 1 in water 3 is brought into contact with at least one cyclodextrin 2. Water is already added in step (a), as a solvent. As a result of the bringing into contact of 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 complexes 12 are present in an aqueous phase.

[0145] FIG. 1 also illustrates the optional continuation of the method according to the invention, where in step (c) the cyclodextrin-AOS complex 12 is separated from the liquid phase by removing water 3. Subsequently, in step (d), the aromatic substance 1 is released from the cyclodextrin-AOS complex 12 by enzymatic treatment. What is obtained is a composition 6 loaded with the apolar organic substance, here the aromatic substance 1. It comprises the concentrated aromatic substance 1 and, as a by-product of the enzymatic treatment, saccharides 20.

[0146] FIG. 2 shows a schematic diagram of a further embodiment of the method according to the invention for selectively separating at least one apolar organic substance (AOS). First, in a step (a), a starting mixture 10 is provided, which contains an ethanolic extract, for example from a solid phase extraction, SPE, in the form of an aromatic substance 1 in an ethanol-containing solution 4 containing 80 vol % of alcohol (ethanol), for example. In step (b), the starting mixture 10 consisting of aromatic substance 1 in ethanolic solution 4 is brought into contact with at least one cyclodextrin 2. As a result of the bringing into contact of 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 ethanol-containing solution 4 of the starting mixture 10. The complexes 12 are present in an aqueous, alcohol-containing phase 4.

[0147] FIG. 2 also illustrates the optional continuation of the method according to the invention, where in step (c) the cyclodextrin-AOS complex 12 is separated from the liquid phase by removing ethanol 40. Subsequently, in step (d), the aromatic substance 1 is released from the cyclodextrin-AOS complex 12 by enzymatic treatment. To this end, water 3 is added in step (c) to the solid phase consisting of the complex 12 of cyclodextrin andin this casearomatic substance, in order to create suitable conditions for the enzymatic treatment. What is obtained from step (d) is a composition 6 loaded with the apolar organic substance, here the aromatic substance 1. It comprises the concentrated aromatic substance 1 and, as a by-product of the enzymatic treatment, saccharides 20. Depending on process control and depending on which ethanol concentrations can be tolerated in the composition for the respective application case, the composition may contain up to 10 vol % of ethanol, for example.

[0148] FIG. 3 shows three photographs, from left to right. They show pictures of a test arrangement consisting of three cylinders. The cylinders are filled with suspensions of cyclodextrin-AOS complexes. Each cylinder on the left contain complexes of gamma-cyclodextrin, each cylinder in the middle contains complexes of beta-cyclodextrin, each cylinder on the right contains complexes in a mixture (right) consisting of 10 wt % beta-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 the beta- 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 different cyclodextrins or the cyclodextrin mixture and were then simultaneously transferred into the graduated cylinders.

[0149] After the start of the comparative experiment at time 0 h (left picture) the sedimentation of the particles from the suspensions can be seen over a duration of 0.5 h (middle picture) up to a duration of 1 h (right picture), showing that the particles from the suspension containing beta-cyclodextrin have already sedimented after 0.5 h and thus twice as fast as those from the suspension containing gamma-cyclodextrin, where a clear supernatant that is visually distinguishable from the sediment at the bottom of the cylinder only appears after a test duration of 1 h.

[0150] Surprisingly, the suspension with the mixture consisting of only 10 wt % beta-cyclodextrin and 90 wt % of gamma-cyclodextrin based on the total mass of cyclodextrin, i.e. predominantly gamma-cyclodextrin, shows the same visual impression of sedimentation behavior as the suspension containing beta-cyclodextrin alone as the cyclodextrin. Thus, according to the invention, an addition of beta-cyclodextrin to gamma-cyclodextrin in a ratio of less than 1:1 is sufficient to increase the settling rate of suspensions with gamma-cyclodextrin to values corresponding to the settling rate of suspensions with beta-cyclodextrin alone.

EXEMPLARY EMBODIMENTS

Example 1 for the Flavoring of, e.g., 0.0 vol % Alcohol Beers

[0151] An ethanol-containing (70 - 80 vol %) solid phase extract [1 kg with an aroma concentration of 4 g/l], which contains isoamyl alcohol, isoamyl acetate, phenylethyl alcohol, hexanoic acid, ethyl hexanoate and other aroma substances, is mixed with 60 g of -, -, and/or -cyclodextrin, preferably with -cyclodextrin. The mixture is stirred for 48 h at 6 C. The aromatic substance-cyclodextrin complex is separated by filtration and dried. To maximize yield, the ethanolic solid phase extract can again be mixed with -, -, and/or -cyclodextrin, preferably -cyclodextrin, namely 30 g per kg of solid phase extract. The alcohol-free filter cake obtained in this way is received in water and treated enzymatically to release the aromatic substance. For this purpose, the aqueous mixture is mixed with 111.1 amylase (Fungamyl from Novozymes) per ml (1 microliter amylase per 1 milliliter of aqueous mixture) and is incubated in a closed vessel for 48 h at 55 C. and a pH of 5.2 (the maximum final cyclodextrin concentration is 0.1 wt % based on the total weight of the aqueous extract). The solubility of the extracted aroma components in water is low, which is why the resulting two-phase mixture can be homogenized with propylene glycol (up to 1:1 w/w), if necessary, and then filtered.

[0152] The so obtained alcohol-free product rich in aromatic substance (5 g/l) can now be used for the flavoring of beers, for example 0.0 vol % alcohol beers (application dosage: 0.2:1000).

[0153] In the context 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; eluent: 10% methanol in water (isocratic); flow: 0.5 ml/min; pressure: 25 bar; oven temperature: 55 C.; run time: 30 minutes).

[0154] The concentration of apolar organic substance, for example the concentration of aromatic substance, of the composition according to the invention, in particular of an alcohol-free extract, is determined by GC-FID (2 g samples were extracted with 2 g cyclohexane. The organic phase is dried over Na2SO4, admixed with internal standard, and analyzed).

Example 2 for the Flavoring of, e.g., 0.0 vol % Alcohol Beers

[0155] A fermentate which contains isoamyl acetate or 4-vinylguaiacol is admixed with a molar equivalent of -cyclodextrin (- and/or -) with respect to the aromatic substance. In order to obtain a stable aromatic substance-cyclodextrin complex, the mixture is stirred for 48 h at 6 C.

[0156] The aromatic substance-cyclodextrin complex is separated by filtration or concentrated on the retentate side.

[0157] Release of the aromatic substance is achieved by an enzymatic treatment of the aromatic substance-cyclodextrin complex. For this purpose, the aqueous mixture was admixed with 111.1 amylase (Fungamyl from Novozymes) per ml (1 microliter amylase per 1 milliliter of aqueous mixture) and was incubated in a closed vessel for 48 h at 55 C. and pH 5.2 (the maximum final cyclodextrin concentration is 0.1 wt % based on the total weight of the aqueous extract). The solubility of the extracted aroma components in water is low, which is why the resulting two-phase mixture can be homogenized with propylene glycol (up to 1:1 w/w) if necessary, and then filtered.

[0158] The so obtained alcohol-free product which is rich in aromatic substance can now be used for the flavoring of beers, for example in 0.0 vol % alcohol beers. The final cyclodextrin concentration is determined by HPLC (detector: RI; separation column: polysep GFC-P 2000 from Phenomenex; eluent: water (isocratic); flow: 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 using GC-FID (2 g samples were extracted with 2 g cyclohexane. The organic phase is dried over Na.sub.2SO.sub.4, admixed with internal standard, and analyzed).

Example 3 for the Flavoring of 0.0 vol % Alcohol Beverages

[0159] A solid phase extract containing ethanol (70-80 vol %) (or a spirit or liqueur (15-96 vol %)) containing isoamyl alcohol, isoamyl acetate, phenylethyl alcohol and other aromatic substances was mixed with 60 g/l of a mixture consisting of beta- and gamma-cyclodextrin (1:9). In order to obtain a sufficiently stable host-guest complex, the mixture was stirred overnight at 6 C. Subsequently, the mixture was allowed to rest for 2 hours for sedimentation (without beta-cyclodextrin, this process may take up to one day).

[0160] After sedimentation, the clear phase was removed and the sediment was dried by filtration.

[0161] For maximizing yield, the ethanolic clear solid phase extract depleted in aromatic substance can again be admixed with a beta- and gamma-cyclodextrin mixture (1:9) and then allowed to rest, having the clear phase removed after sedimentation, and then being subjected to filtration.

[0162] The alcohol-free filter cake obtained in this way was received in water and treated enzymatically to release the aromatic substance. For this purpose, the aqueous mixture was admixed with 1 l amylase Dextrozyme GA (Novozymes) per milliliter and incubated in a closed vessel for 48 h at 55 C. and pH 4.5. Finally, the resulting mixture was filtered.

[0163] The so obtained alcohol-free product which is rich in aromatic substance can now be used in low doses, for example in a range from 0.01:1000 to 50:1000, for flavoring purposes, for example to flavor 0.0 vol % alcohol beers.

Example 4 for the Extraction of Apple Wax-Containing Fraction from Dried Apple Pomace

[0164] 1600 g of dry apple pomace was admixed with 8000 g of a 36 wt % aqueous cyclodextrin solution (beta-cyclodextrin to gamma-cyclodextrin in a 1:1 ratio, i.e. 18 g beta- and 18 g gamma-cyclodextrin to 1 liter of water), and was kneaded for 2 days at room temperature. The wet apple pomace was then filtered off, and the filtrate rich in cyclodextrin-apple wax complex was collected as a suspension.

[0165] As step (c) of the method according to the invention, the suspension was centrifuged, and the clear phase was decanted off

[0166] The so obtained extract (182.65 g) was admixed with 200 g of water and incubated with the amylase Dextrozyme GA (400 l) for 48 h at a pH of 4.5 and 55 C. The mixture was then filtered off using vacuum, and was dried to constant weight on a rotary evaporator. The so obtained fraction rich in apple wax (16 g) was successfully tested as a constituent of a release agent for fruit gums, which means that the fraction rich in apple wax obtained in this way can be used as a release agent just as well as a release agent containing a wax (also apple wax) that was obtained through others previously known methods.

Example 5 for the Extraction of Aroma-Containing Oil Fraction from Hop Cones

[0167] 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 at room temperature for 2 days. The wet hop cones were then filtered off, and the filtrate rich in cyclodextrin-oil complex was collected as a suspension.

[0168] As step (c) of the method according to the invention, the suspension was centrifuged and the clear phase was decanted off.

[0169] The so obtained extract (28.99 g) was admixed with 15 g of water and incubated with the amylase Dextrozyme GA (88 l) for 48 h at a pH of 4.5 and 55 C. The aroma-rich extract obtained in this way has a high aroma load, allowing the extract obtained in this way to be used in a dosage of 1:1000 for flavoring, for example, alcohol-free beer.

Example 6 for the Extraction of Aromatic Substances from Ethanol-Containing Gentian Root Extract

[0170] An extract of gentian roots with an alcohol content of 60 vol % was mixed with 6 wt % of gamma-cyclodextrin in solid form. The mixture was stored overnight at 6 C. In this way, sufficiently stable hostguest complexes were obtained.

[0171] The host-guest complex was separated by filtration as step (c) of the method according to the invention. Water was added to the filter cake. Release of the aromatic substances was achieved by enzymatic treatment of the host-guest complex. For this purpose, the aqueous mixture was admixed with 1 l of the amylase Dextrozyme GA (Novozymes) per milliliter and incubated in a closed vessel for 48 h at 55 C. Finally, the resulting mixture was filtered.

[0172] The aroma-rich product obtained can now be used in low dosage for flavoring purposes, for example in products with an alcohol content of 0.0 vol % ethanol.

[0173] It will be apparent to a person skilled in the art that the invention is not limited to the examples described above, but rather can be varied in a variety of ways. The features of the individually illustrated examples can in particular also be combined with one another or interchanged with one another.

LIST OF REFERENCE NUMERALS

[0174] 1 Organic substance comprising at least one apolar group, for short: apolar organic substance (AOS); aromatic substance, bitter substance, oil, fat, wax, adhesive, colorant

[0175] 10 Starting mixture

[0176] 2 Cyclodextrin

[0177] 12 Guesthost complex of cyclodextrin and organic substance comprising at least one apolar group (AOS), cyclodextrinAOS complex

[0178] 20 Saccharides

[0179] 3 Water

[0180] 4 Ethanol-containing solution

[0181] 40 Ethanol

[0182] 5 Enzyme

[0183] 6 Composition