CA2948964C - Method for obtaining sinapic acid from a native material mixture - Google Patents
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- CA2948964C CA2948964C CA2948964A CA2948964A CA2948964C CA 2948964 C CA2948964 C CA 2948964C CA 2948964 A CA2948964 A CA 2948964A CA 2948964 A CA2948964 A CA 2948964A CA 2948964 C CA2948964 C CA 2948964C
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/48—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/05—Mashed or comminuted pulses or legumes; Products made therefrom
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/028—Flow sheets
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0292—Treatment of the solvent
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/04—Solvent extraction of solutions which are liquid
- B01D11/0492—Applications, solvents used
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/47—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
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Abstract
A method is described for obtaining sinapic acid and a salt of sinapic acid from a mixture of seed and fruits having hard frangible husks. The process involves comminuting and dispersing with water. Agitation results in a free flowing pulp. The pulp pH is set to an alkaline range greater than 9.5.
A water-soluble organic solvent is added to detach husks from endosperm of the seeds and fruits. The pH of the resulting pulp is then shifted to a range of 4.5 to 7.2 and the pulp is separated into a plurality of phases. An extraction step follows in which at least one of the sinapic acid and a derivative of sinapic acid is isolated from the pulp. The process optimizes obtaining valuable materials from the mixture and provides a relatively simple process for obtaining a sinapic acid or derivatives of sinapic acid.
A water-soluble organic solvent is added to detach husks from endosperm of the seeds and fruits. The pH of the resulting pulp is then shifted to a range of 4.5 to 7.2 and the pulp is separated into a plurality of phases. An extraction step follows in which at least one of the sinapic acid and a derivative of sinapic acid is isolated from the pulp. The process optimizes obtaining valuable materials from the mixture and provides a relatively simple process for obtaining a sinapic acid or derivatives of sinapic acid.
Description
METHOD FOR OBTAINING SINAPIC ACID FROM A NATIVE MATERIAL MIXTURE
The present invention relates to a method for obtaining sinapic acid and/or a salt of sinapic acid from a native material mixture.
It is known to obtain a protein phase as a valuable material phase from seeds having hard frangible husks, in particular from oil seed rape fruits.
In the familiar approach for protein concentrate production, the grain meal (greatly deoiled) is washed, wherein the soluble extracted materials are depleted. The value of the deoiled intermediates depends greatly on the concentration of accompanying materials, such as fibers, sugars and secondary plant materials (Menner, M. et al. "Fraktionierung pflanzlicher Rohstoffe zur simultanen Erzeugung von Lebensmitteln, technischen Rohstoffen und Energietragern" [Fractionation of plant raw materials for simultaneous production of foods, technical raw materials and energy carriers], Chemie Ingenieur Technik, volume 81, edition 11, pages 1743 - 1756, November 2009). These accompanying materials also include polyphenols such as sinapine. The polyphenol acid "sinapic acid" occurs primarily in rape seeds (there the sinapine content is approximately 640 mg/100 g of oil seed rape). In order to separate off accompanying materials such as sinapine, large dilutions are chosen, proteins are also denatured (temperature, alcohol), cellulose is enzymatically broken down to form short-chain carbohydrates; these methods are chosen in order to be able to extract the materials better.
The present invention relates to a method for obtaining sinapic acid and/or a salt of sinapic acid from a native material mixture.
It is known to obtain a protein phase as a valuable material phase from seeds having hard frangible husks, in particular from oil seed rape fruits.
In the familiar approach for protein concentrate production, the grain meal (greatly deoiled) is washed, wherein the soluble extracted materials are depleted. The value of the deoiled intermediates depends greatly on the concentration of accompanying materials, such as fibers, sugars and secondary plant materials (Menner, M. et al. "Fraktionierung pflanzlicher Rohstoffe zur simultanen Erzeugung von Lebensmitteln, technischen Rohstoffen und Energietragern" [Fractionation of plant raw materials for simultaneous production of foods, technical raw materials and energy carriers], Chemie Ingenieur Technik, volume 81, edition 11, pages 1743 - 1756, November 2009). These accompanying materials also include polyphenols such as sinapine. The polyphenol acid "sinapic acid" occurs primarily in rape seeds (there the sinapine content is approximately 640 mg/100 g of oil seed rape). In order to separate off accompanying materials such as sinapine, large dilutions are chosen, proteins are also denatured (temperature, alcohol), cellulose is enzymatically broken down to form short-chain carbohydrates; these methods are chosen in order to be able to extract the materials better.
- 2 -Against this background, the problem addressed by the invention is to further optimize obtaining valuable materials from the native material mixture, wherein it is to be possible, in particular, to obtain in a relatively simple manner a sinapine, a sinapic acid or derivatives of sinapic acid.
SUMMARY
Accordingly, there is described a method for obtaining at least one of sinapic acid and a salt of sinapic acid from a native material mixture comprising the following steps:
- step A): providing a native material mixture of seeds/fruits having hard frangible husks comprising oil seed rape fruits of the Brassicaceae family;
- step B): if the native material mixture of step A is not yet comminuted: comminuting the native material mixture;
- step C): dispersing the comminuted native material mixture of step A) or B) with water, wherein, to one part of comminuted native material mixture up to a maximum of 8 parts of water are added, and wherein the water and the native comminuted material mixture are agitated such that one of a free-flowing pulp and a dispersion results;
- step D): setting the pH of the pulp from step C) to an alkaline range pH > 9.5;
- step E): adding a water-soluble organic solvent, to the pulp from step C) subsequent to the setting of the pH of the pulp in step D) in order to detach the husks from endosperm of the seeds and fruits;
- step F): separating off a solids phase which has the predominant fraction of the husks;
- step G): shifting the pH of the pulp which is freed from husks from step F) to the pH range from pH = 4.5 to pH = 7.2;
Date Recue/Date Received 2022-01-18 - 2a -- step H): separating the husk-free pulp, the pH of which has been shifted to the acidic range in step G) into a plurality of phases, wherein at least one of said plurality of phases is a polyphenol-albumin liquid phase; and - step I): isolating at least one of the sinapic acid and the salt of sinapic acid from the husk-free pulp of step F) or G) or from the polyphenol-albumin liquid phase of step H) directly or after the husk-free pulp passes through further intermediate steps, by an extraction with an extraction medium.
BRIEF DESCRIPTION OF THE DRAWINGS
Aspects of the present invention are illustrated, merely by way of example, in the following drawings in which:
Figures la and lb illustrate different batches of different raw materials or starting materials plus water used in embodiments of the current process, wherein the batch amounts are normalized;
Figure 2 is a schematic diagram showing the overall process according to an embodiment.
In more detail, in an embodiment, the steps of the described process involve:
Step A:
As starting material, a native material mixture is provided from seeds having hard frangible husks, in particular from - seeds/fruits of crucifers (Brassicaceae), in particular of oil seed rape fruits.
Date Recue/Date Received 2022-01-18 - 2b -The material mixture within the meaning of this application can consist of the complete, but broken, seeds. These can be unhusked or partially husked or completely husked.
Alternatively, the material mixture can also consist of a previously deoiled product, in particular of an "intermediate", that is to say of a press cake which remains as residue of obtaining oil after a "preliminary stage", e.g. after pressing off oil, in particular using a press (e.g. a screw press).
Date Recue/Date Received 2022-01-18
SUMMARY
Accordingly, there is described a method for obtaining at least one of sinapic acid and a salt of sinapic acid from a native material mixture comprising the following steps:
- step A): providing a native material mixture of seeds/fruits having hard frangible husks comprising oil seed rape fruits of the Brassicaceae family;
- step B): if the native material mixture of step A is not yet comminuted: comminuting the native material mixture;
- step C): dispersing the comminuted native material mixture of step A) or B) with water, wherein, to one part of comminuted native material mixture up to a maximum of 8 parts of water are added, and wherein the water and the native comminuted material mixture are agitated such that one of a free-flowing pulp and a dispersion results;
- step D): setting the pH of the pulp from step C) to an alkaline range pH > 9.5;
- step E): adding a water-soluble organic solvent, to the pulp from step C) subsequent to the setting of the pH of the pulp in step D) in order to detach the husks from endosperm of the seeds and fruits;
- step F): separating off a solids phase which has the predominant fraction of the husks;
- step G): shifting the pH of the pulp which is freed from husks from step F) to the pH range from pH = 4.5 to pH = 7.2;
Date Recue/Date Received 2022-01-18 - 2a -- step H): separating the husk-free pulp, the pH of which has been shifted to the acidic range in step G) into a plurality of phases, wherein at least one of said plurality of phases is a polyphenol-albumin liquid phase; and - step I): isolating at least one of the sinapic acid and the salt of sinapic acid from the husk-free pulp of step F) or G) or from the polyphenol-albumin liquid phase of step H) directly or after the husk-free pulp passes through further intermediate steps, by an extraction with an extraction medium.
BRIEF DESCRIPTION OF THE DRAWINGS
Aspects of the present invention are illustrated, merely by way of example, in the following drawings in which:
Figures la and lb illustrate different batches of different raw materials or starting materials plus water used in embodiments of the current process, wherein the batch amounts are normalized;
Figure 2 is a schematic diagram showing the overall process according to an embodiment.
In more detail, in an embodiment, the steps of the described process involve:
Step A:
As starting material, a native material mixture is provided from seeds having hard frangible husks, in particular from - seeds/fruits of crucifers (Brassicaceae), in particular of oil seed rape fruits.
Date Recue/Date Received 2022-01-18 - 2b -The material mixture within the meaning of this application can consist of the complete, but broken, seeds. These can be unhusked or partially husked or completely husked.
Alternatively, the material mixture can also consist of a previously deoiled product, in particular of an "intermediate", that is to say of a press cake which remains as residue of obtaining oil after a "preliminary stage", e.g. after pressing off oil, in particular using a press (e.g. a screw press).
Date Recue/Date Received 2022-01-18
3 PCT/EP2015/061644 Particularly preferably, the starting material processed is "intermediate obtained shortly before", that is to say after the preliminary stage no more than 31 days must have passed.
The seed can be freshly harvested or else be days, weeks or months old, the intermediate stage (the pressing) should take place shortly before, or even immediately before, the further processing in order that, after obtaining the oil, the material - the seed - has not changed too greatly.
Highly preferably, the starting material processed is "fresh material", i.e., after a preliminary stage, or preliminary processing (obtaining oil), no more than three days must have passed, preferably even only fewer than 48 hours or 24 hours or 12 hours or fewer than 1 hour.
Using material from a time period shortly after the preliminary stage, good results are achieved or, and using fresh material, generally even better results are achieved with respect to the yield and purity of the products of value.
The press cake can have a residual oil content that can even be 20% or more. Despite such high residual oil contents, even obtaining a protein phase using the invention is achievable in a simple manner. Obtaining the protein, however, is only optional.
The sinapic acid and/or the sinapine salt can therefore be obtained as the sole product of value of the material mixture, or as an additional product of value during obtaining of protein.
Step B:
If it is not yet comminuted: comminuting the material mixture of step a) to break open the husks. If a press cake is used, said press cake is broken open, ideally immediately after pressing, still warm. In this manner, a comminuted material - of granule type - is generated from the press cake. The material mixture that is (partially) deoiled beforehand by a pressing operation is generally only comminuted, for example crumbled or granulated, or at all events the husks are broken open.
Step C:
13 The material mixture that is provided and comminuted from step A) or B) is dispersed with water or an aqueous solution (e.g. a salt solution). To one part of "comminuted product", preferably up to a maximum of 8, preferably up to a maximum of 5, parts (weight fractions) of water are added. Then, water and comminuted product are stirred, such that a free-flowing pulp or a dispersion results. The stirring preferably proceeds for more than 30 min, in particular for more than 1 h.
Step D) Next, the pH of the pulp (I) from step c) is set to an alkaline range; preferably, the pH of the pulp or of the dispersion is set to pH 10 to 11 using alkali metal hydroxide solution. In this case, the stirring is continued (carefully). The stirring time is preferably more than 30 min, preferably it is 1 h or more.
Step E) In this further step, at least one water-soluble organic solvent, preferably alcohol, is added, in particular in water-diluted form, subsequently to setting the pH of the pulp in step D. Preferably, the dispersion, the pH of which has been set to the alkaline range, is brought to an alcohol concentration of 20-15% by volume or less, using the alcohol Et0H (preferably 30-60%), in particular a concentration of 12%
Et0H. Corresponding to the amount of water of the alcohol used, the amount of water in step C can be reduced by the water present in the alcohol, in particular in the 30-60% Et0H. The husks thereby detach from the endosperm (cotyledon) with the residual oil and can be separated off, in particular centrifugally.
As less preferred alternative in comparison with ethanol, other alcohols, such as, e.g., isopropanol, can also be used.
The steps C-E can proceed together, for example simultaneously in time. Thus, it is, e.g., possible to set an aqueous highly dilute ethanol solution using alkali metal hydroxide solution to a correspondingly basic pH and to add this solution to the comminuted material mixture.
Step F) In step F), therefore, a solids phase is separated off, which comprises the predominant fraction of the husks, preferably in a centrifuge in the centrifugal field, from the pulp, or the pulp is clarified from husk and solids fractions, in particular in a decanter.
In this step, the husks are separated from the residual pulp using a decanter having a feed.
The lighter phase of a centrifugal phase separation will hereinafter also occasionally be termed as overflow, and the solids phase as heavy phase. A middle phase may correspondingly lie inbetween, relating to the density thereof.
Step G) The at all events substantially husk-free pulp from step F) is then further processed. Preferably, in this case, the dissolved protein fraction is precipitated out of the husk-free pulp, which protein fraction, together with the non-dissolved or solubilized protein portion, forms a fraction, the curd. The pH
in this case is shifted back to the acidic range, in particular to the pH range from pH = 4.5 to pH = 7.
Step H) Then, the husk-free pulp, the pH of which has been shifted back to the acidic range, is separated - preferably in a centrifuge, in particular in at least one decanter or in a separator - in one or two steps into valuable material phases, of which one phase is a protein concentrate phase and one of said phases is a polyphenol-albumin liquid phase.
Particularly preferably, a separation into the following two or three phases takes place:
- oil-containing phase - aqueous phase (polyphenol- and sinapic acid-containing) - protein concentrate phase (hereinafter also called "protein curd") or - aqueous phase with albumin and polyphenol contents and residual oil content; and - protein concentrate phase (protein curd).
The two-phase separation is carried out when the raw material is relatively highly deoiled and/or is present in bound form in the solids, or if an intensive shearing effect has not been implemented for the liquid phase in step 1. The addition of water or alcohol or alkali metal hydroxide solution or the like can also proceed in substeps. The oil as a lighter phase contains triglycerides and is one of the valuable materials that may be obtained.
Step I): Isolating sinapic acid or the sinapines from the husk-free pulp of step F) or - and this is particularly preferred - from the polyphenol-albumin liquid phase of step H) directly or after passage through further intermediate steps, in particular by an extraction using an extraction medium, in particular after addition of an extraction medium to the husk-free pulp of step F) or C). In particular, ethyl acetate is suitable, which will be demonstrated hereinafter with reference to experiments. However, pentanol is also usable.
Hereinafter, mostly sinapic acid and sinapic acid-containing phases and material mixtures are mentioned. Of course, however, depending on the pH, sinapic acid can also be present as sinapic acid esterified with choline.
Preferably, the temperature during all of the method steps is below 60 C, in particular below 50 C, preferably between 40 C
and 50 C, as a result of which particularly valuable products may be obtained.
The denaturation of the proteins is a temperature- and time-dependent process. In addition, there is the condition in the alcoholic milieu. The protein denaturation proceeds the more rapidly the higher the temperature is. In an aqueous environment, however, under the action of heat of 45-50 C, irreversible protein denaturation is also not to be expected.
However, this changes with the alcohol concentration. Even at ambient temperature, in highly concentrated alcohol, a protein precipitation may be observed.
The lower then the alcohol concentration is, the higher must the temperature be in order to denature the proteins. Or vica versa:
the more aqueous the alcohol concentration is, the higher may the working temperature be without the proteins being irreversibly damaged.
Therefore, (for pure water) a temperature that is as high as possible, that is to say as far as possible reaching to 60 C, will be chosen in order to bring as many substances as possible into solution, such as proteins, lecithins, glycolipids, etc.
The precipitated proteins are present as protein curd (heavy phase).
The advantageous temperature data for the method steps A to H
does not relate to the pressing temperature during generation of the press cake in the oil production. The higher the temperature was in the preceding process steps, the browner the protein phase or curd fraction becomes. This firstly concerns the Maillard reaction of sugars with proteins, and secondly phenol oxidation. In comparison with DE 10 2011 050 905 Al, in particular owing to the use of optimally selected starting materials (preferably cold-pressed oil seed rape press cake, preferably very fresh), a particularly attractive, particularly readily reusable product is obtained.
The use of cold-pressed materials (in particular a cold-pressed oil seed rape press cake; temperature during pressing advantageously below 70 C, particularly preferably even below 60 C) as starting material or as the material mixture provided is particularly advantageous. Warm-pressed material is exposed during pressing to markedly higher temperatures (up to above 100 C). By using cold-pressed material as starting material for the method according to the invention, a protein phase or "protein and/or curd phase" having markedly improved properties (in particular with respect to the color markedly brighter and therefore more readily processable) and having markedly improved yield can be obtained than is the case when warm- or hot-pressed starting material is used. This has not been acknowledged to date in the prior art. This is because customary oil seed rape pressing methods are targeted at a high oil yield, for which reason, during pressing, relatively high temperatures are willingly used. As a side effect, it may be observed that sinapine (a polyphenol) is broken down, which in itself would seem to be advantageous for the protein fraction. In the method according to the invention, the original, that is to say non-reduced, sinapine content in the cold-pressed cake, however, nevertheless is not a problem for the end product, since the polyphenolic compounds are substantially not recovered in the curd phase, since they transfer into the water phase.
In this case, the curd phase which was obtained by the method according to the invention from a press cake previously additionally deoiled with hexane is rather to be assigned to the RAL color 1024 ochre yellow or 1014 ivory or a mixture of these two colors. The processing preferably proceeds at ambient pressure.
In the liquid phase and/or "water phase" of step H), valuable constituents are still also present, in particular it has a relatively high albumin content. To this extent, it would be logical and advantageous to perform a filtration of the water phase for albumin concentration, in order in this manner to obtain the albumin phase as a further valuable material.
As a further valuable constituent, sinapic acid and/or sinapine and/or polyphenols may be obtained from the liquid phase and/or "water phase" of step H).
This is possible in a simple manner in that the sinapic acid is extracted from the aqueous phase using an extraction medium that is suitable therefor. As suitable extraction medium, in particular ethyl acetate and pentanol are suitable, wherein preference is given to ethyl acetate.
Alternatively, it is conceivable to perform the extraction of sinapic acid directly from the liquid phase - the protein-containing pulp - of step F) using an extraction medium that is suitable therefor.
A particularly advantageous method variant may be explained with reference to the following example.
Step A) The starting material provided in this example is pressed oil seed rape cake, ideally pressed under mild and cold conditions, having typical residual oil contents of 20%; but higher contents are not a problem.
Step B) The cake is broken open, ideally directly after pressing, still warm.
Step C) The cake granules are dispersed with water (1 part cake and a maximum of 6 parts water) and are to be stirred carefully (1 h).
Step D) This dispersion is to be set to pH 10 to 11 using alkali metal hydroxide solution and carefully stirred, usually for 1 h.
Step E) The dispersion from 4 is to be brought to an Et0H
concentration of 12% by volume using Et0H (preferably 30-60% - based on percent by volume); the amount of water in point 3. is thereby reduced by the water present in said 30-60%
Et0H.
Step F) The husks thereby detach from the endosperm (cotyledon) with the residual oil and can be separated off centrifugally.
Step G) Precipitation of the protein by acidification to preferably pH = 4.5 to 7.2 out of the overflow (overflow: light phase of the separation from step S6 having a pH from preferably 9.7 to 10.5) for the separation: oil - aqueous phase - protein concentrate phase (protein curd) or for separation into oil/water phase and protein concentrate phase; this step can be supported by intensive shearing in order to facilitate the oil release.
Step H) Separating off the precipitated proteins as curd (heavy phase (generally solids phase, or here curd phase)) and optionally triglycerides (as light oil) from the overflow (light phase), in particular centrifugally.
Step I) Filtration of the water phase for albumin concentration.
The wet separation of the husks from the dissolved and swollen proteins in a parallel displacement extraction of the triglycerides (oil phase) from oil-containing or residual oil-containing press cake or legume meal and parallel polyphenol extraction may be mentioned as particularly advantageous.
The particular advantages of the method are:
Low dilutions and thus small volumetric streams are achievable in the process via the above-described method with simultaneously low solvent waste.
This gives a higher polyphenol concentration during the extraction in the aqueous phase (method steps 2 to 5).
Native temperature-sensitive proteins are in addition present in the end product, since the process is implemented at a maximum of 50-55 C or below.
Overall, comparatively high protein yields of up to 75% by weight are achievable, wherein up to 45-50% by weight can be obtained from the "curd phase" and approximately 22-24% by weight from the albumin phase.
A relatively high-quality end product (protein mixture) can be obtained, because husk residues, and also polyphenols, carbohydrates, lignin, cellulose, etc. are completely removed or depleted.
The protein phase contains "native" protein, the swellable fraction of which remains swellable after it is obtained, and the water-dissolvable fractions of which remain water-soluble after they are obtained. The protein phase is in addition virtually triglyceride-free and has only low residual oil values, principally polar lipids.
The good milieu for microorganism growth due to the slight alcohol concentration simplifies the process hygiene.
The alcohol can be circulated in dilute form.
With regard to steps A) and B):
Instead of the extraction of unwanted materials from the highly deoiled, very finely comminuted starting material oil seed rape meal or oil seed rape cake - as is usual in the familiar methods - here first the husks are separated off in the wet state. This is solved in a multistage process in that first the cake is broken up, without the kernel fragments being further comminuted.
In particular, it is important to leave the husks as large as possible. Preferably, they should have a mean diameter of 0.5 mm or greater. Oil droplets do not need to be greater; what is of importance is not individual molecules or small molecule aggregates, but "particles".
Then, water is added and the mixture is carefully stirred in alkaline conditions. The water-soluble part of the proteins is dissolved thereby, another part swells. The addition of aqueous alcohol displaces the free triglyceride as a specific light gravity phase out of the dispersion. The lecithins, in particular phosphatidyl cholines, are soluble at low alcohol concentrations (see EP 1272048 Bl and associated patent family).
In this combination of alkali metal hydroxide solution - aqueous alcohol, the two or three phases are 1) Heavy = husks and 2) light = protein-lecithin-polyphenol-carbohydrate together with oil-containing foam; or 1) Heavy = husks, 2) medium - protein-lecithin-polyphenol-carbohydrate, 3) light = triglyceride, advantageously separable, for instance in the experiment in the glass beaker or on an industrial scale, preferably by centrifugation.
The better the husks are separated off, the lower are the protein losses and the purer is the end product. Even the husk swollen up to 7 times by addition of water is heavier than the proteins in the alcoholic-aqueous dispersion. This is essential for separation by gravitation. However, the separation is made more difficult by firm adhesion of the protein-containing aleurone bodies (honeycomb layer) to the husks. These cells are thick-walled. Since the cell membrane of virtually all cells contains lecithins (in addition to proteins and other substances), the adhesion can now be minimized by suitable measures by "solubilizing" of the lecithins.
Specifically, this is achieved by the aqueous phase having an alcohol concentration of 5-40% by volume (see steps S2-S4), ideally 12% to 20%.
The quality of the starting material is critical therefor. The residual oil content is usually higher in the case of cold-pressed cake. This does not interfere in the case of the method presented here. On the contrary: the gentle pressing is extremely helpful; the more moderate is the pressing temperature and the lower the pressing force, the easier is subsequent separation of husks and cotyledon (germ layers, the kernel interior).
The method is also applicable with "usual", that is to say hot-pressed, press cake. Only in this case are the yields of proteins correspondingly lower.
Regarding steps C) to E) (dispersion production):
Producing the dispersion with water-aqueous alkali metal hydroxide solution and alcohol has two aims: firstly, the detachment from the husk, secondly the extraction of phenolic compounds such as sinapine from the raw material. In this case, the wetting with fluid is of importance. However, a shearing in the case of the dispersion formulation in steps 2-5 generated very small particles that led to impurities in the separated phases. Without use of a shear-head mixer, or without using a toothed-wheel mixer for steps 2-5, the protein content in the protein phase (after the step in which it is obtained and a drying) was: > 60% by weight in the case of fresh material.
Using a shear-head mixer, or using a Frisam shear mixer, the protein content in the protein phase (the step in which it is obtained and a drying) was: approximately 50% in the case of old material.
Another test was carried out using hot-pressed expeller cake.
The amount of removable husks in the first stage is decreased by the shearing from 20% to 16%, and at the same time the amount of precipitable protein from the clear phase is increased from 38%
by weight to 42% by weight. The purity remains relatively constant and low at 39-40% by weight.
The water-aqueous alkali metal hydroxide solution and alcohol dispersion is stirred at a temperature of T = 50 C for about 30 min.
In addition to dissolution of the lecithins in the aqueous-alcoholic solution, for improved separation of husks firstly, and triglycerides secondly, separating off the husks in the slightly alcoholic solution has the additional advantage that growth of microorganisms in the process is made more difficult. This is a marked advantage in comparison with the purely aqueous method and facilitates the CIP cleaning.
Regarding step F) separation The separation will be described hereinafter with reference to some examples for better illustration.
Example:
El) A cold-pressed protein-containing cake which is processed as far as step F, after processing thereof, has the following phases: 17% heavy phase as husk fraction from the feed with 20%
of the cake proteins and 83% overflow as protein-polyphenol-oil-phosphatide phase having 80% of the cake proteins.
B2) A warm-pressed cake, which is processed up to step F, has, after processing thereof, the following phases: 26% heavy phase as husk fraction from the feed having 30% of the cake proteins and 74% overflow as protein-polyphenol-oil-phosphatide phase having 70% of the cake proteins.
B3) A hot-pressed cake, which is processed in step F, after processing thereof has the following phases: 30% heavy phase as husk fraction from the feed having 50% of the cake proteins and 70% overflow as protein-polyphenol-oil-phosphatide phase having 50% of the cake proteins.
Regarding step G) - protein precipitation The proteins are precipitated out of the overflow (overflow - light phase) of the separation in the preceding step by pH shifting to the range from 4.5 to approximately 7. The water-insoluble proteins which, however, are swellable in aqueous solution form, together with the precipitated globulins, the protein fraction of the "protein curd". The liquid in this fraction has the same composition as the liquid of the middle phase (overflow without triglycerides). Since, however, the curd phase only makes up 10-30% by weight of the feed, (having a relatively high fraction of dry matter, 15-25% by weight of dry matter), quantitatively, substantially fewer polyphenols may also be found in the curd phase than in the middle phase, even if the concentration of the polyphenols, based on the water, is the same.
Therefore, a protein phase of water-insoluble but swollen proteins with globulins is available, which is depleted with polyphenols. This combination of alkaline-ethanolic milieu in steps A-F, followed by an acid-alcohol milieu for protein precipitation, represents very good conditions for a polyphenol extraction. Surprisingly, for oil seed rape (sinapine and derivatives), here the observation for other polyphenols (tyrosol and derivatives, inter alia) from other fields such as the processing of olives is confirmed, although significantly more reactive substances such as proteins and sugars are present in the suspension.
Therefore, dilutions such as described in the literature are irrelevant in order to arrive at equivalent polyphenol extraction rates of the aqueous mixture (for instance, again Kroll et al., "Rapssamenproteine - Struktur, Eigenschaften, Gewinnung und Modifizierung" [Oil seed rape proteins -structure, properties, production and modification], Deutsche Lebensmittel-Rundschau, volume 3, 2007, p. 109).
Since the pure triglyceride is displaced from the liquid as a light phase, the residual oil content in the protein end product can be reduced to below 15% by weight, also below 13% by weight, based on dry matter.
Since the temperatures during the entire process are <= 50 C, this can also be described as a native end product.
If the pulp that is to be processed further is sheared before the phase separation of step H (before the oil separation) and after step F) or G) of claim 1, this is advantageous for improving the displacement extraction. This shearing can be carried out using a shearing device such as, e.g., a homogenizer, or an intensive mixer, in order to obtain still more oil in this manner.
The shearing can be carried out using a shearing device in a continuous process. Overall, preferably, a continuous process is implemented.
Regarding step H) - separation of the proteins as curd by means of decanter or separator To increase the purity, the protein curd can be washed. The curd can then be dried to form a powder.
Regarding step I) - isolating the sinapic acid, the sinapine or a derivative of sinapic acid Subsequently, advantageously sinapic acid is obtained by means of an extraction and optionally an albumin phase is obtained (the latter, for example, by filtration).
In short, an advantageous method for obtaining proteins from native material mixtures is also provided, having the following steps:
A) providing a native material mixture of seeds having hard frangible husks, B) breaking up or coarsely comminuting the material mixture in order in any case to break open the husks without dispersing them too finely. Preferably, the size of the comminuted husk fractions in a granulometric distribution by type should be between 100 and 2000 pm, in particular having a maximum between 300 pm and 900 pm, in particular approximately 600 pm, in each case at a relative frequency of greater than 5%;
C) dispersing the comminuted broken-open material mixture from step A) or B) with water or an aqueous solution;
D) setting the pH of the pulp (I) from step C) to an alkaline range of pH > 9,5;
E) adding the water-soluble organic solvent alcohol to the pulp of step C) subsequently to setting the pH of the pulp in step D;
F) separating off a solids phase which has the predominant fraction of the husks, preferably in a centrifuge in the centrifugal field;
G) shifting the pH of the pulp freed from husks from step F) to the pH range from pH = 4.5 to pH = 7.2, and H) separating the husk-free pulp, the pH of which has been shifted back to the acidic range - preferably in a centrifuge, in particular in at least one decanter - in one or two steps into the following three valuable material phases: oil-containing phase having a triglyercol content; aqueous phase having an albumin content and protein concentrate phase (protein curd).
When the oil content is very low in the raw material, the oil phase is absent and only two phases are formed, the aqueous albumin phase and the protein concentrate phase.
It is further advantageous if the curd phase is dried. Here it is advantageous to vaporize alcohol still present out of the curd, preferably under vacuum, in order to keep the temperature low and to dry the alcohol-free aqueous curd to form a powder.
For this purpose, for example, a drying and a grinding are advantageous; for implementation thereof in an apparatus, the drier-pulverizer is suitable. In this manner, a storage-stable, readily handleable and also transportable product is provided.
The beneficial properties can be illustrated with reference to a protein phase obtained in the experiment:
Experiment 1 (regarding steps A-F): experimental batch: 95 kg of tap water + 23 kg of oil seed rape (warm pressing) were charged into an agitator reservoir and heated to 40 C (steps A and C).
Subsequently, this product/water mixture was circulated by means of a monopump and a Fristam mixer at 1000 l/h for approximately 8 min (pH = 6.2). Then 4.1 1 of 10% NaOH are added and the pH is set to 10 (step D).
The mixture was subsequently circulated without the Fristam mixer at 1000 l/h for 15 min and stirred. Then 14.2 kg of ethanol are added (step E in one or more substeps) by means of a peristaltic pump directly to the circuit of the monopump.
Residence time: preferably 10-50 min. After approximately 50 min residence time, again 2 kg of ethanol in 11 kg of water are mixed and added to an agitator vessel. 10 min residence time.
This suspension is separated centrifugally (step F). In this case the yield is: 96.5 kg of clear phase, 34 kg of solids. The husks may be readily separated off thereby.
Experiment 2: (A-F) experimental batch: 116 kg of tap water and 26 kg of oil seed rape (cold pressing) were charged into the agitator reservoir and heated to 40 C. Subsequently, the mixture was circulated (pH = 5.8) by means of monopump and Fristam mixer at 1000 l/h for approximately 8 min. Addition of 4.5 1 of 10%
NaOH to pH = 10. Subsequently, the mixture was circulated for 15 min without the Fristam mixer at 1000 1/h. Then 17.2 kg of ethanol were added by means of a peristaltic pump directly into the circuit of the monopump. There is a 10 minute residence time. Thereafter the mixture is separated in order to separate off the husk fraction. The yield is yield: 129.6 kg of clear phase, 26.5 kg of solids. The husks may be more readily separated off thereby.
Experiment 3: (steps G and H to experiment 1): 96.5 kg of clear phase from experiment 1 (starting pH: 9.6) were shifted to pH 5 by means of 0.8 1 of 25% hydrochloric acid, here advantageously at 45 C (step G). This pulp can then be centrifuged, wherein a protein phase is obtained as heavy phase or solids phase (step H). Yield of clear phase: 64.3 kg. Yield of solids phase (curd-type) 9 kg.
Experiment 4 (steps G and H to experiment 2): 129.6 kg of clear phase from experiment II (starting pH: 9.5) were shifted to pH 5 by means of 1.2 1 of 25% hydrochloric acid at 45 C (step G).
This pulp could then be centrifuged, wherein a protein phase was obtained as heavy phase or solids phase (step H). Yield: 83 kg of clear phase, 29.5 kg of solids/protein phase. Here the yield of solids phase is particularly high.
Typically, a powder obtained from a curd obtained in the manner of the abovementioned experiments and then dried has dry matter contents of 5-9%; in a sample produced from conventional press cake 5.35%. The protein content is approximately 60%. The water-binding capacity was determined at 1.8 +/- 0.2 ml H20 per 1 g of dry matter of the protein powder, the oil-binding capacity at 0.49 to 0.63 g of oil per 1 g of dry matter, and also the emulsifiability at 700 to 780 ml of oil per 1 g of dry matter of the protein powder. Typical values for the protein solubility index NSI are 9 to 16%. The best results are achieved with the cold-pressed press cake.
In further experiments it was surprisingly found that the stirring technique in the method is also of importance, which relates, in particular, to the stirring of step C) (and optionally D) and E)) of claim 1.
Thus, cold-pressed oil seed rape press cake was processed in the procedure described in claim 1. In this case, in step C), stirring was performed once using a blade agitator and once using a propeller agitator.
The blade agitator should be operated in such a manner that it generates the fewest possible shearing forces during the stirring but generates a substantially uniform laminar flow.
In the case of the propeller agitator in the meaning of this application, stirring elements are also connected outside the axis of rotation, thus, via a disk or a ring or in the vicinity thereof, elements such as an open bell are present over the propeller elements. They therefore generate a relatively turbulent flow during stirring and exert higher shearing forces on the product.
Blade agitators are therefore those which substantially generate a laminar flow during stirring, which have relatively long blades and which are operated at a low speed of rotation. A ring or a disk or the like on the outer periphery of the blades or in the vicinity thereof (in the manner of an open cage or an open bell around the blades) is generally not present.
In the further experiments, the husk-containing solids phase according to step F) was separated off after steps D) and E) -preferably with further stirring using the blade agitator or the propeller agitator.
The liquid phase after separating off the husks from a suspension of oil seed rape press cake, which contained 13.4%
oil, 31.4% protein and 55.2% others (such as cellulose, polyphenols, saccharides, etc.), when the blade agitator was used for stirring in step C) and optionally D) and E), was markedly protein-richer than when a propeller agitator was used.
Approximately 75% of the proteins of the cake were found in this overflow, the dry matter of which is composed of 52.3% protein, 13.0% oil and approximately 34% others. In contrast, only 62.5%
of the proteins of the cake were found in the comparable overflow, when a propeller agitator was used. For this case, the overflow dry matter had only approximately 37% protein, approximately 14.7% oil and also 48.0% other constituents.
Surprisingly, marked differences were also found visually. The husk fraction of the centrifuge sample from the suspension using the blade agitator appeared markedly marbled. In this case only 42% of the dry matter was separated off as overflow; in the case of the propeller agitator, the fraction of the dry matter separated off was 50%.
Furthermore, still more advantageous method variants could be found.
Thus, a high alcohol concentration, in particular ethanol concentration, causes a high oil content in the "globulin curd".
It is particularly advantageous in step E), therefore, when the alcohol concentration is less than 20% by volume, in particular 13 to 18% by volume, particularly preferably 15% by volume.
An excessively long reaction time at pH 10 (overnight) likewise causes high oil contents in the globulin curd. Somewhat lower temperatures, in particular below 43 C, act advantageously in the globulin precipitation and globulin separation, and give rise to higher protein contents in the curd.
In addition, one or more of the following further measures appear to be particularly advantageous: the use of fresh materials during pressing of the oil; cold pressing of the oil and a gentle stirring with a blade agitator (in step C), wherein the material should be sheared or ground as little as possible.
In addition, an advantageous alcohol content, in particular ethanol content, of less than 20% is particularly advantageous, since otherwise a higher oil content results in the curd.
For the obtaining of sinapic acid of step I), experiments have likewise been carried out.
Thus, it has been found that in a pretreatment of steps C), D) and E), the sinapic acid is enriched in the "water phase". It is therefore obtainable by extraction using a suitable extraction medium, more precisely either after step H) or even after step F).
In this case, however, the choice of starting material also has an effect on the amount of obtainable sinapic acid.
For this purpose, reference may be made to the diagrams of the accompanying figures la and b. In the individual experiments, different batches of different raw material or starting material plus water were selected, wherein the samples, although they contained different amounts, the amount was normalized or suitably converted.
The two diagrams show that the polyphenol content in the aqueous phase can increase to more than 4-fold if, as starting material, "cold-pressed oil seed rape press cake" is used instead of "hot-pressed oil seed rape press cake". The use of fresh material is also advantageous to this extent. This is because, just as the polyphenol fraction is enriched in the water phase, it is depleted in the protein curd phase. Thus, in the case of a hot-pressed cake, the fraction of 9.4% polyphenols (dry matter "DM" in the raw material) is depleted to 5.6% by weight DM in the protein curd or curd powder, or in the cold-pressed cake from 18.6% by weight DM to 10.1% by weight DM in the curd powder. Therefore, this concentration of the polyphenols based on the dry matter in the solids is only about half as great as in the starting material.
Therefore, a protein phase of water-insoluble but swollen proteins with globulins is available that has been depleted with respect to the polyphenol content. In the water phase, approximately 55-55% by weight of the polyphenols remain in the following concentrations:
Cake type Dilution in PP In the PP in the the method water phase water phase (mg) normalized Parts of to a water based dilution 1 on 1 part of part seed +
cake 6 parts fluid Cold 4.5 3976 2982 Warm 4.2 3183 2228 Hot 6.0 1053 1058 The following influence factors should be heeded in the processing: in the hot pressing, polyphenols (PP) are broken down. It has been measured that the PP content in clear-pressed seed was 18 mg/g, but in the case of hot-pressed seed was 8.8 mg/g. Similar values are known from the literature (6.2 mg/g in Jeroch et al. 1999). In addition to the reduction of the polyphenols in the raw material, the sinapine is deesterified to form sinapic acid.
Via the cold pressing, according to the abovementioned method, the polyphenols are transferred most highly in terms of quantity in the cold pressing into the water phase or more precisely "polyphenol-albumin phase" of step H). They are present, as a result of the alkaline pretreatment (+ temperature and Et0H), substantially as sinapic acid or salts of sinapic acid, and no longer as sinapine and not yet as canolol.
It is henceforth advantageous to extract the sinapic acid in one step I) from the aqueous phase using an extraction medium, in particular ethyl acetate. For this purpose, the polyphenol-albumin phase was washed with ethyl acetate, or alternatively with pentanol.
a) with 1/1 (PP-albumin phase/ethyl acetate) b) with 1/2 (PP-albumin phase/ethyl acetate) c) with 1/1 (PP-albumin phase/pentanol) d) with 1/2 (PP-albumin-phase/pentanol) The once-extracted polyphenol phase as valuable material phase of stages a) to d) was in each case washed a further time (2nd extraction). This was performed with the original pH from 5.2 to 5.4, and repeated once more at pH 4. This gave the following table of results:
Extraction Extraction Extraction Extraction rate in % rate at pH rate at pH rate at pH
by weight 5.2 and 4 4 and 2nd at pH 5.2 2nd stage stage a ethyl 74.6 78.2 72.5 90.8 acetate ethyl 57.8 77.0 82.7 91.1 acetate 2nd wash 74.6 84.8 85.6 87.4 pentanol 61.1 80.7 85.5 92.1 pentanol 2nd wash Extraction with ethyl acetate appears to be particularly advantageous. This is again advantageously added in the aqueous mixture in the ratio 1/0.5 to 1/3 (quantitative ratio:
PP-albumin phase/ethyl acetate). The ethyl acetate can be vaporized at the end.
The above-described steps are again clearly shown in fig. 2.
The seed can be freshly harvested or else be days, weeks or months old, the intermediate stage (the pressing) should take place shortly before, or even immediately before, the further processing in order that, after obtaining the oil, the material - the seed - has not changed too greatly.
Highly preferably, the starting material processed is "fresh material", i.e., after a preliminary stage, or preliminary processing (obtaining oil), no more than three days must have passed, preferably even only fewer than 48 hours or 24 hours or 12 hours or fewer than 1 hour.
Using material from a time period shortly after the preliminary stage, good results are achieved or, and using fresh material, generally even better results are achieved with respect to the yield and purity of the products of value.
The press cake can have a residual oil content that can even be 20% or more. Despite such high residual oil contents, even obtaining a protein phase using the invention is achievable in a simple manner. Obtaining the protein, however, is only optional.
The sinapic acid and/or the sinapine salt can therefore be obtained as the sole product of value of the material mixture, or as an additional product of value during obtaining of protein.
Step B:
If it is not yet comminuted: comminuting the material mixture of step a) to break open the husks. If a press cake is used, said press cake is broken open, ideally immediately after pressing, still warm. In this manner, a comminuted material - of granule type - is generated from the press cake. The material mixture that is (partially) deoiled beforehand by a pressing operation is generally only comminuted, for example crumbled or granulated, or at all events the husks are broken open.
Step C:
13 The material mixture that is provided and comminuted from step A) or B) is dispersed with water or an aqueous solution (e.g. a salt solution). To one part of "comminuted product", preferably up to a maximum of 8, preferably up to a maximum of 5, parts (weight fractions) of water are added. Then, water and comminuted product are stirred, such that a free-flowing pulp or a dispersion results. The stirring preferably proceeds for more than 30 min, in particular for more than 1 h.
Step D) Next, the pH of the pulp (I) from step c) is set to an alkaline range; preferably, the pH of the pulp or of the dispersion is set to pH 10 to 11 using alkali metal hydroxide solution. In this case, the stirring is continued (carefully). The stirring time is preferably more than 30 min, preferably it is 1 h or more.
Step E) In this further step, at least one water-soluble organic solvent, preferably alcohol, is added, in particular in water-diluted form, subsequently to setting the pH of the pulp in step D. Preferably, the dispersion, the pH of which has been set to the alkaline range, is brought to an alcohol concentration of 20-15% by volume or less, using the alcohol Et0H (preferably 30-60%), in particular a concentration of 12%
Et0H. Corresponding to the amount of water of the alcohol used, the amount of water in step C can be reduced by the water present in the alcohol, in particular in the 30-60% Et0H. The husks thereby detach from the endosperm (cotyledon) with the residual oil and can be separated off, in particular centrifugally.
As less preferred alternative in comparison with ethanol, other alcohols, such as, e.g., isopropanol, can also be used.
The steps C-E can proceed together, for example simultaneously in time. Thus, it is, e.g., possible to set an aqueous highly dilute ethanol solution using alkali metal hydroxide solution to a correspondingly basic pH and to add this solution to the comminuted material mixture.
Step F) In step F), therefore, a solids phase is separated off, which comprises the predominant fraction of the husks, preferably in a centrifuge in the centrifugal field, from the pulp, or the pulp is clarified from husk and solids fractions, in particular in a decanter.
In this step, the husks are separated from the residual pulp using a decanter having a feed.
The lighter phase of a centrifugal phase separation will hereinafter also occasionally be termed as overflow, and the solids phase as heavy phase. A middle phase may correspondingly lie inbetween, relating to the density thereof.
Step G) The at all events substantially husk-free pulp from step F) is then further processed. Preferably, in this case, the dissolved protein fraction is precipitated out of the husk-free pulp, which protein fraction, together with the non-dissolved or solubilized protein portion, forms a fraction, the curd. The pH
in this case is shifted back to the acidic range, in particular to the pH range from pH = 4.5 to pH = 7.
Step H) Then, the husk-free pulp, the pH of which has been shifted back to the acidic range, is separated - preferably in a centrifuge, in particular in at least one decanter or in a separator - in one or two steps into valuable material phases, of which one phase is a protein concentrate phase and one of said phases is a polyphenol-albumin liquid phase.
Particularly preferably, a separation into the following two or three phases takes place:
- oil-containing phase - aqueous phase (polyphenol- and sinapic acid-containing) - protein concentrate phase (hereinafter also called "protein curd") or - aqueous phase with albumin and polyphenol contents and residual oil content; and - protein concentrate phase (protein curd).
The two-phase separation is carried out when the raw material is relatively highly deoiled and/or is present in bound form in the solids, or if an intensive shearing effect has not been implemented for the liquid phase in step 1. The addition of water or alcohol or alkali metal hydroxide solution or the like can also proceed in substeps. The oil as a lighter phase contains triglycerides and is one of the valuable materials that may be obtained.
Step I): Isolating sinapic acid or the sinapines from the husk-free pulp of step F) or - and this is particularly preferred - from the polyphenol-albumin liquid phase of step H) directly or after passage through further intermediate steps, in particular by an extraction using an extraction medium, in particular after addition of an extraction medium to the husk-free pulp of step F) or C). In particular, ethyl acetate is suitable, which will be demonstrated hereinafter with reference to experiments. However, pentanol is also usable.
Hereinafter, mostly sinapic acid and sinapic acid-containing phases and material mixtures are mentioned. Of course, however, depending on the pH, sinapic acid can also be present as sinapic acid esterified with choline.
Preferably, the temperature during all of the method steps is below 60 C, in particular below 50 C, preferably between 40 C
and 50 C, as a result of which particularly valuable products may be obtained.
The denaturation of the proteins is a temperature- and time-dependent process. In addition, there is the condition in the alcoholic milieu. The protein denaturation proceeds the more rapidly the higher the temperature is. In an aqueous environment, however, under the action of heat of 45-50 C, irreversible protein denaturation is also not to be expected.
However, this changes with the alcohol concentration. Even at ambient temperature, in highly concentrated alcohol, a protein precipitation may be observed.
The lower then the alcohol concentration is, the higher must the temperature be in order to denature the proteins. Or vica versa:
the more aqueous the alcohol concentration is, the higher may the working temperature be without the proteins being irreversibly damaged.
Therefore, (for pure water) a temperature that is as high as possible, that is to say as far as possible reaching to 60 C, will be chosen in order to bring as many substances as possible into solution, such as proteins, lecithins, glycolipids, etc.
The precipitated proteins are present as protein curd (heavy phase).
The advantageous temperature data for the method steps A to H
does not relate to the pressing temperature during generation of the press cake in the oil production. The higher the temperature was in the preceding process steps, the browner the protein phase or curd fraction becomes. This firstly concerns the Maillard reaction of sugars with proteins, and secondly phenol oxidation. In comparison with DE 10 2011 050 905 Al, in particular owing to the use of optimally selected starting materials (preferably cold-pressed oil seed rape press cake, preferably very fresh), a particularly attractive, particularly readily reusable product is obtained.
The use of cold-pressed materials (in particular a cold-pressed oil seed rape press cake; temperature during pressing advantageously below 70 C, particularly preferably even below 60 C) as starting material or as the material mixture provided is particularly advantageous. Warm-pressed material is exposed during pressing to markedly higher temperatures (up to above 100 C). By using cold-pressed material as starting material for the method according to the invention, a protein phase or "protein and/or curd phase" having markedly improved properties (in particular with respect to the color markedly brighter and therefore more readily processable) and having markedly improved yield can be obtained than is the case when warm- or hot-pressed starting material is used. This has not been acknowledged to date in the prior art. This is because customary oil seed rape pressing methods are targeted at a high oil yield, for which reason, during pressing, relatively high temperatures are willingly used. As a side effect, it may be observed that sinapine (a polyphenol) is broken down, which in itself would seem to be advantageous for the protein fraction. In the method according to the invention, the original, that is to say non-reduced, sinapine content in the cold-pressed cake, however, nevertheless is not a problem for the end product, since the polyphenolic compounds are substantially not recovered in the curd phase, since they transfer into the water phase.
In this case, the curd phase which was obtained by the method according to the invention from a press cake previously additionally deoiled with hexane is rather to be assigned to the RAL color 1024 ochre yellow or 1014 ivory or a mixture of these two colors. The processing preferably proceeds at ambient pressure.
In the liquid phase and/or "water phase" of step H), valuable constituents are still also present, in particular it has a relatively high albumin content. To this extent, it would be logical and advantageous to perform a filtration of the water phase for albumin concentration, in order in this manner to obtain the albumin phase as a further valuable material.
As a further valuable constituent, sinapic acid and/or sinapine and/or polyphenols may be obtained from the liquid phase and/or "water phase" of step H).
This is possible in a simple manner in that the sinapic acid is extracted from the aqueous phase using an extraction medium that is suitable therefor. As suitable extraction medium, in particular ethyl acetate and pentanol are suitable, wherein preference is given to ethyl acetate.
Alternatively, it is conceivable to perform the extraction of sinapic acid directly from the liquid phase - the protein-containing pulp - of step F) using an extraction medium that is suitable therefor.
A particularly advantageous method variant may be explained with reference to the following example.
Step A) The starting material provided in this example is pressed oil seed rape cake, ideally pressed under mild and cold conditions, having typical residual oil contents of 20%; but higher contents are not a problem.
Step B) The cake is broken open, ideally directly after pressing, still warm.
Step C) The cake granules are dispersed with water (1 part cake and a maximum of 6 parts water) and are to be stirred carefully (1 h).
Step D) This dispersion is to be set to pH 10 to 11 using alkali metal hydroxide solution and carefully stirred, usually for 1 h.
Step E) The dispersion from 4 is to be brought to an Et0H
concentration of 12% by volume using Et0H (preferably 30-60% - based on percent by volume); the amount of water in point 3. is thereby reduced by the water present in said 30-60%
Et0H.
Step F) The husks thereby detach from the endosperm (cotyledon) with the residual oil and can be separated off centrifugally.
Step G) Precipitation of the protein by acidification to preferably pH = 4.5 to 7.2 out of the overflow (overflow: light phase of the separation from step S6 having a pH from preferably 9.7 to 10.5) for the separation: oil - aqueous phase - protein concentrate phase (protein curd) or for separation into oil/water phase and protein concentrate phase; this step can be supported by intensive shearing in order to facilitate the oil release.
Step H) Separating off the precipitated proteins as curd (heavy phase (generally solids phase, or here curd phase)) and optionally triglycerides (as light oil) from the overflow (light phase), in particular centrifugally.
Step I) Filtration of the water phase for albumin concentration.
The wet separation of the husks from the dissolved and swollen proteins in a parallel displacement extraction of the triglycerides (oil phase) from oil-containing or residual oil-containing press cake or legume meal and parallel polyphenol extraction may be mentioned as particularly advantageous.
The particular advantages of the method are:
Low dilutions and thus small volumetric streams are achievable in the process via the above-described method with simultaneously low solvent waste.
This gives a higher polyphenol concentration during the extraction in the aqueous phase (method steps 2 to 5).
Native temperature-sensitive proteins are in addition present in the end product, since the process is implemented at a maximum of 50-55 C or below.
Overall, comparatively high protein yields of up to 75% by weight are achievable, wherein up to 45-50% by weight can be obtained from the "curd phase" and approximately 22-24% by weight from the albumin phase.
A relatively high-quality end product (protein mixture) can be obtained, because husk residues, and also polyphenols, carbohydrates, lignin, cellulose, etc. are completely removed or depleted.
The protein phase contains "native" protein, the swellable fraction of which remains swellable after it is obtained, and the water-dissolvable fractions of which remain water-soluble after they are obtained. The protein phase is in addition virtually triglyceride-free and has only low residual oil values, principally polar lipids.
The good milieu for microorganism growth due to the slight alcohol concentration simplifies the process hygiene.
The alcohol can be circulated in dilute form.
With regard to steps A) and B):
Instead of the extraction of unwanted materials from the highly deoiled, very finely comminuted starting material oil seed rape meal or oil seed rape cake - as is usual in the familiar methods - here first the husks are separated off in the wet state. This is solved in a multistage process in that first the cake is broken up, without the kernel fragments being further comminuted.
In particular, it is important to leave the husks as large as possible. Preferably, they should have a mean diameter of 0.5 mm or greater. Oil droplets do not need to be greater; what is of importance is not individual molecules or small molecule aggregates, but "particles".
Then, water is added and the mixture is carefully stirred in alkaline conditions. The water-soluble part of the proteins is dissolved thereby, another part swells. The addition of aqueous alcohol displaces the free triglyceride as a specific light gravity phase out of the dispersion. The lecithins, in particular phosphatidyl cholines, are soluble at low alcohol concentrations (see EP 1272048 Bl and associated patent family).
In this combination of alkali metal hydroxide solution - aqueous alcohol, the two or three phases are 1) Heavy = husks and 2) light = protein-lecithin-polyphenol-carbohydrate together with oil-containing foam; or 1) Heavy = husks, 2) medium - protein-lecithin-polyphenol-carbohydrate, 3) light = triglyceride, advantageously separable, for instance in the experiment in the glass beaker or on an industrial scale, preferably by centrifugation.
The better the husks are separated off, the lower are the protein losses and the purer is the end product. Even the husk swollen up to 7 times by addition of water is heavier than the proteins in the alcoholic-aqueous dispersion. This is essential for separation by gravitation. However, the separation is made more difficult by firm adhesion of the protein-containing aleurone bodies (honeycomb layer) to the husks. These cells are thick-walled. Since the cell membrane of virtually all cells contains lecithins (in addition to proteins and other substances), the adhesion can now be minimized by suitable measures by "solubilizing" of the lecithins.
Specifically, this is achieved by the aqueous phase having an alcohol concentration of 5-40% by volume (see steps S2-S4), ideally 12% to 20%.
The quality of the starting material is critical therefor. The residual oil content is usually higher in the case of cold-pressed cake. This does not interfere in the case of the method presented here. On the contrary: the gentle pressing is extremely helpful; the more moderate is the pressing temperature and the lower the pressing force, the easier is subsequent separation of husks and cotyledon (germ layers, the kernel interior).
The method is also applicable with "usual", that is to say hot-pressed, press cake. Only in this case are the yields of proteins correspondingly lower.
Regarding steps C) to E) (dispersion production):
Producing the dispersion with water-aqueous alkali metal hydroxide solution and alcohol has two aims: firstly, the detachment from the husk, secondly the extraction of phenolic compounds such as sinapine from the raw material. In this case, the wetting with fluid is of importance. However, a shearing in the case of the dispersion formulation in steps 2-5 generated very small particles that led to impurities in the separated phases. Without use of a shear-head mixer, or without using a toothed-wheel mixer for steps 2-5, the protein content in the protein phase (after the step in which it is obtained and a drying) was: > 60% by weight in the case of fresh material.
Using a shear-head mixer, or using a Frisam shear mixer, the protein content in the protein phase (the step in which it is obtained and a drying) was: approximately 50% in the case of old material.
Another test was carried out using hot-pressed expeller cake.
The amount of removable husks in the first stage is decreased by the shearing from 20% to 16%, and at the same time the amount of precipitable protein from the clear phase is increased from 38%
by weight to 42% by weight. The purity remains relatively constant and low at 39-40% by weight.
The water-aqueous alkali metal hydroxide solution and alcohol dispersion is stirred at a temperature of T = 50 C for about 30 min.
In addition to dissolution of the lecithins in the aqueous-alcoholic solution, for improved separation of husks firstly, and triglycerides secondly, separating off the husks in the slightly alcoholic solution has the additional advantage that growth of microorganisms in the process is made more difficult. This is a marked advantage in comparison with the purely aqueous method and facilitates the CIP cleaning.
Regarding step F) separation The separation will be described hereinafter with reference to some examples for better illustration.
Example:
El) A cold-pressed protein-containing cake which is processed as far as step F, after processing thereof, has the following phases: 17% heavy phase as husk fraction from the feed with 20%
of the cake proteins and 83% overflow as protein-polyphenol-oil-phosphatide phase having 80% of the cake proteins.
B2) A warm-pressed cake, which is processed up to step F, has, after processing thereof, the following phases: 26% heavy phase as husk fraction from the feed having 30% of the cake proteins and 74% overflow as protein-polyphenol-oil-phosphatide phase having 70% of the cake proteins.
B3) A hot-pressed cake, which is processed in step F, after processing thereof has the following phases: 30% heavy phase as husk fraction from the feed having 50% of the cake proteins and 70% overflow as protein-polyphenol-oil-phosphatide phase having 50% of the cake proteins.
Regarding step G) - protein precipitation The proteins are precipitated out of the overflow (overflow - light phase) of the separation in the preceding step by pH shifting to the range from 4.5 to approximately 7. The water-insoluble proteins which, however, are swellable in aqueous solution form, together with the precipitated globulins, the protein fraction of the "protein curd". The liquid in this fraction has the same composition as the liquid of the middle phase (overflow without triglycerides). Since, however, the curd phase only makes up 10-30% by weight of the feed, (having a relatively high fraction of dry matter, 15-25% by weight of dry matter), quantitatively, substantially fewer polyphenols may also be found in the curd phase than in the middle phase, even if the concentration of the polyphenols, based on the water, is the same.
Therefore, a protein phase of water-insoluble but swollen proteins with globulins is available, which is depleted with polyphenols. This combination of alkaline-ethanolic milieu in steps A-F, followed by an acid-alcohol milieu for protein precipitation, represents very good conditions for a polyphenol extraction. Surprisingly, for oil seed rape (sinapine and derivatives), here the observation for other polyphenols (tyrosol and derivatives, inter alia) from other fields such as the processing of olives is confirmed, although significantly more reactive substances such as proteins and sugars are present in the suspension.
Therefore, dilutions such as described in the literature are irrelevant in order to arrive at equivalent polyphenol extraction rates of the aqueous mixture (for instance, again Kroll et al., "Rapssamenproteine - Struktur, Eigenschaften, Gewinnung und Modifizierung" [Oil seed rape proteins -structure, properties, production and modification], Deutsche Lebensmittel-Rundschau, volume 3, 2007, p. 109).
Since the pure triglyceride is displaced from the liquid as a light phase, the residual oil content in the protein end product can be reduced to below 15% by weight, also below 13% by weight, based on dry matter.
Since the temperatures during the entire process are <= 50 C, this can also be described as a native end product.
If the pulp that is to be processed further is sheared before the phase separation of step H (before the oil separation) and after step F) or G) of claim 1, this is advantageous for improving the displacement extraction. This shearing can be carried out using a shearing device such as, e.g., a homogenizer, or an intensive mixer, in order to obtain still more oil in this manner.
The shearing can be carried out using a shearing device in a continuous process. Overall, preferably, a continuous process is implemented.
Regarding step H) - separation of the proteins as curd by means of decanter or separator To increase the purity, the protein curd can be washed. The curd can then be dried to form a powder.
Regarding step I) - isolating the sinapic acid, the sinapine or a derivative of sinapic acid Subsequently, advantageously sinapic acid is obtained by means of an extraction and optionally an albumin phase is obtained (the latter, for example, by filtration).
In short, an advantageous method for obtaining proteins from native material mixtures is also provided, having the following steps:
A) providing a native material mixture of seeds having hard frangible husks, B) breaking up or coarsely comminuting the material mixture in order in any case to break open the husks without dispersing them too finely. Preferably, the size of the comminuted husk fractions in a granulometric distribution by type should be between 100 and 2000 pm, in particular having a maximum between 300 pm and 900 pm, in particular approximately 600 pm, in each case at a relative frequency of greater than 5%;
C) dispersing the comminuted broken-open material mixture from step A) or B) with water or an aqueous solution;
D) setting the pH of the pulp (I) from step C) to an alkaline range of pH > 9,5;
E) adding the water-soluble organic solvent alcohol to the pulp of step C) subsequently to setting the pH of the pulp in step D;
F) separating off a solids phase which has the predominant fraction of the husks, preferably in a centrifuge in the centrifugal field;
G) shifting the pH of the pulp freed from husks from step F) to the pH range from pH = 4.5 to pH = 7.2, and H) separating the husk-free pulp, the pH of which has been shifted back to the acidic range - preferably in a centrifuge, in particular in at least one decanter - in one or two steps into the following three valuable material phases: oil-containing phase having a triglyercol content; aqueous phase having an albumin content and protein concentrate phase (protein curd).
When the oil content is very low in the raw material, the oil phase is absent and only two phases are formed, the aqueous albumin phase and the protein concentrate phase.
It is further advantageous if the curd phase is dried. Here it is advantageous to vaporize alcohol still present out of the curd, preferably under vacuum, in order to keep the temperature low and to dry the alcohol-free aqueous curd to form a powder.
For this purpose, for example, a drying and a grinding are advantageous; for implementation thereof in an apparatus, the drier-pulverizer is suitable. In this manner, a storage-stable, readily handleable and also transportable product is provided.
The beneficial properties can be illustrated with reference to a protein phase obtained in the experiment:
Experiment 1 (regarding steps A-F): experimental batch: 95 kg of tap water + 23 kg of oil seed rape (warm pressing) were charged into an agitator reservoir and heated to 40 C (steps A and C).
Subsequently, this product/water mixture was circulated by means of a monopump and a Fristam mixer at 1000 l/h for approximately 8 min (pH = 6.2). Then 4.1 1 of 10% NaOH are added and the pH is set to 10 (step D).
The mixture was subsequently circulated without the Fristam mixer at 1000 l/h for 15 min and stirred. Then 14.2 kg of ethanol are added (step E in one or more substeps) by means of a peristaltic pump directly to the circuit of the monopump.
Residence time: preferably 10-50 min. After approximately 50 min residence time, again 2 kg of ethanol in 11 kg of water are mixed and added to an agitator vessel. 10 min residence time.
This suspension is separated centrifugally (step F). In this case the yield is: 96.5 kg of clear phase, 34 kg of solids. The husks may be readily separated off thereby.
Experiment 2: (A-F) experimental batch: 116 kg of tap water and 26 kg of oil seed rape (cold pressing) were charged into the agitator reservoir and heated to 40 C. Subsequently, the mixture was circulated (pH = 5.8) by means of monopump and Fristam mixer at 1000 l/h for approximately 8 min. Addition of 4.5 1 of 10%
NaOH to pH = 10. Subsequently, the mixture was circulated for 15 min without the Fristam mixer at 1000 1/h. Then 17.2 kg of ethanol were added by means of a peristaltic pump directly into the circuit of the monopump. There is a 10 minute residence time. Thereafter the mixture is separated in order to separate off the husk fraction. The yield is yield: 129.6 kg of clear phase, 26.5 kg of solids. The husks may be more readily separated off thereby.
Experiment 3: (steps G and H to experiment 1): 96.5 kg of clear phase from experiment 1 (starting pH: 9.6) were shifted to pH 5 by means of 0.8 1 of 25% hydrochloric acid, here advantageously at 45 C (step G). This pulp can then be centrifuged, wherein a protein phase is obtained as heavy phase or solids phase (step H). Yield of clear phase: 64.3 kg. Yield of solids phase (curd-type) 9 kg.
Experiment 4 (steps G and H to experiment 2): 129.6 kg of clear phase from experiment II (starting pH: 9.5) were shifted to pH 5 by means of 1.2 1 of 25% hydrochloric acid at 45 C (step G).
This pulp could then be centrifuged, wherein a protein phase was obtained as heavy phase or solids phase (step H). Yield: 83 kg of clear phase, 29.5 kg of solids/protein phase. Here the yield of solids phase is particularly high.
Typically, a powder obtained from a curd obtained in the manner of the abovementioned experiments and then dried has dry matter contents of 5-9%; in a sample produced from conventional press cake 5.35%. The protein content is approximately 60%. The water-binding capacity was determined at 1.8 +/- 0.2 ml H20 per 1 g of dry matter of the protein powder, the oil-binding capacity at 0.49 to 0.63 g of oil per 1 g of dry matter, and also the emulsifiability at 700 to 780 ml of oil per 1 g of dry matter of the protein powder. Typical values for the protein solubility index NSI are 9 to 16%. The best results are achieved with the cold-pressed press cake.
In further experiments it was surprisingly found that the stirring technique in the method is also of importance, which relates, in particular, to the stirring of step C) (and optionally D) and E)) of claim 1.
Thus, cold-pressed oil seed rape press cake was processed in the procedure described in claim 1. In this case, in step C), stirring was performed once using a blade agitator and once using a propeller agitator.
The blade agitator should be operated in such a manner that it generates the fewest possible shearing forces during the stirring but generates a substantially uniform laminar flow.
In the case of the propeller agitator in the meaning of this application, stirring elements are also connected outside the axis of rotation, thus, via a disk or a ring or in the vicinity thereof, elements such as an open bell are present over the propeller elements. They therefore generate a relatively turbulent flow during stirring and exert higher shearing forces on the product.
Blade agitators are therefore those which substantially generate a laminar flow during stirring, which have relatively long blades and which are operated at a low speed of rotation. A ring or a disk or the like on the outer periphery of the blades or in the vicinity thereof (in the manner of an open cage or an open bell around the blades) is generally not present.
In the further experiments, the husk-containing solids phase according to step F) was separated off after steps D) and E) -preferably with further stirring using the blade agitator or the propeller agitator.
The liquid phase after separating off the husks from a suspension of oil seed rape press cake, which contained 13.4%
oil, 31.4% protein and 55.2% others (such as cellulose, polyphenols, saccharides, etc.), when the blade agitator was used for stirring in step C) and optionally D) and E), was markedly protein-richer than when a propeller agitator was used.
Approximately 75% of the proteins of the cake were found in this overflow, the dry matter of which is composed of 52.3% protein, 13.0% oil and approximately 34% others. In contrast, only 62.5%
of the proteins of the cake were found in the comparable overflow, when a propeller agitator was used. For this case, the overflow dry matter had only approximately 37% protein, approximately 14.7% oil and also 48.0% other constituents.
Surprisingly, marked differences were also found visually. The husk fraction of the centrifuge sample from the suspension using the blade agitator appeared markedly marbled. In this case only 42% of the dry matter was separated off as overflow; in the case of the propeller agitator, the fraction of the dry matter separated off was 50%.
Furthermore, still more advantageous method variants could be found.
Thus, a high alcohol concentration, in particular ethanol concentration, causes a high oil content in the "globulin curd".
It is particularly advantageous in step E), therefore, when the alcohol concentration is less than 20% by volume, in particular 13 to 18% by volume, particularly preferably 15% by volume.
An excessively long reaction time at pH 10 (overnight) likewise causes high oil contents in the globulin curd. Somewhat lower temperatures, in particular below 43 C, act advantageously in the globulin precipitation and globulin separation, and give rise to higher protein contents in the curd.
In addition, one or more of the following further measures appear to be particularly advantageous: the use of fresh materials during pressing of the oil; cold pressing of the oil and a gentle stirring with a blade agitator (in step C), wherein the material should be sheared or ground as little as possible.
In addition, an advantageous alcohol content, in particular ethanol content, of less than 20% is particularly advantageous, since otherwise a higher oil content results in the curd.
For the obtaining of sinapic acid of step I), experiments have likewise been carried out.
Thus, it has been found that in a pretreatment of steps C), D) and E), the sinapic acid is enriched in the "water phase". It is therefore obtainable by extraction using a suitable extraction medium, more precisely either after step H) or even after step F).
In this case, however, the choice of starting material also has an effect on the amount of obtainable sinapic acid.
For this purpose, reference may be made to the diagrams of the accompanying figures la and b. In the individual experiments, different batches of different raw material or starting material plus water were selected, wherein the samples, although they contained different amounts, the amount was normalized or suitably converted.
The two diagrams show that the polyphenol content in the aqueous phase can increase to more than 4-fold if, as starting material, "cold-pressed oil seed rape press cake" is used instead of "hot-pressed oil seed rape press cake". The use of fresh material is also advantageous to this extent. This is because, just as the polyphenol fraction is enriched in the water phase, it is depleted in the protein curd phase. Thus, in the case of a hot-pressed cake, the fraction of 9.4% polyphenols (dry matter "DM" in the raw material) is depleted to 5.6% by weight DM in the protein curd or curd powder, or in the cold-pressed cake from 18.6% by weight DM to 10.1% by weight DM in the curd powder. Therefore, this concentration of the polyphenols based on the dry matter in the solids is only about half as great as in the starting material.
Therefore, a protein phase of water-insoluble but swollen proteins with globulins is available that has been depleted with respect to the polyphenol content. In the water phase, approximately 55-55% by weight of the polyphenols remain in the following concentrations:
Cake type Dilution in PP In the PP in the the method water phase water phase (mg) normalized Parts of to a water based dilution 1 on 1 part of part seed +
cake 6 parts fluid Cold 4.5 3976 2982 Warm 4.2 3183 2228 Hot 6.0 1053 1058 The following influence factors should be heeded in the processing: in the hot pressing, polyphenols (PP) are broken down. It has been measured that the PP content in clear-pressed seed was 18 mg/g, but in the case of hot-pressed seed was 8.8 mg/g. Similar values are known from the literature (6.2 mg/g in Jeroch et al. 1999). In addition to the reduction of the polyphenols in the raw material, the sinapine is deesterified to form sinapic acid.
Via the cold pressing, according to the abovementioned method, the polyphenols are transferred most highly in terms of quantity in the cold pressing into the water phase or more precisely "polyphenol-albumin phase" of step H). They are present, as a result of the alkaline pretreatment (+ temperature and Et0H), substantially as sinapic acid or salts of sinapic acid, and no longer as sinapine and not yet as canolol.
It is henceforth advantageous to extract the sinapic acid in one step I) from the aqueous phase using an extraction medium, in particular ethyl acetate. For this purpose, the polyphenol-albumin phase was washed with ethyl acetate, or alternatively with pentanol.
a) with 1/1 (PP-albumin phase/ethyl acetate) b) with 1/2 (PP-albumin phase/ethyl acetate) c) with 1/1 (PP-albumin phase/pentanol) d) with 1/2 (PP-albumin-phase/pentanol) The once-extracted polyphenol phase as valuable material phase of stages a) to d) was in each case washed a further time (2nd extraction). This was performed with the original pH from 5.2 to 5.4, and repeated once more at pH 4. This gave the following table of results:
Extraction Extraction Extraction Extraction rate in % rate at pH rate at pH rate at pH
by weight 5.2 and 4 4 and 2nd at pH 5.2 2nd stage stage a ethyl 74.6 78.2 72.5 90.8 acetate ethyl 57.8 77.0 82.7 91.1 acetate 2nd wash 74.6 84.8 85.6 87.4 pentanol 61.1 80.7 85.5 92.1 pentanol 2nd wash Extraction with ethyl acetate appears to be particularly advantageous. This is again advantageously added in the aqueous mixture in the ratio 1/0.5 to 1/3 (quantitative ratio:
PP-albumin phase/ethyl acetate). The ethyl acetate can be vaporized at the end.
The above-described steps are again clearly shown in fig. 2.
Claims
EMBODIMENTS IN WHICH AN EXCLUSIVE PROPERTY OR PRIVILEGE IS
CLAIMED ARE DEFINED AS FOLLOWS:
1.
A method for obtaining at least one of sinapic acid and a salt of sinapic acid from a native material mixture comprising the following steps:
- step A): providing a native material mixture of seeds/fruits having hard frangible husks comprising oil seed rape fruits of the Brassicaceae family;
- step B): if the native material mixture of step A is not yet comminuted: comminuting the native material mixture;
- step C): dispersing the comminuted native material mixture of step A) or B) with water, wherein, to one part of comminuted native material mixture up to a maximum of 8 parts of water are added, and wherein the water and the comminuted native material mixture are agitated such that one of a free-flowing pulp and a dispersion results;
- step D): setting the pH of the pulp from step C) to an alkaline range pH > 9.5;
- step E): adding a water-soluble organic solvent, to the pulp from step C) subsequent to the setting of the pH of the pulp in step D) in order to detach the husks from endosperm of the seeds and fruits;
- step F): separating off a solids phase which has the predominant fraction of the husks;
- step G): shifting the pH of the pulp which is freed from husks from step F) to the pH range from pH = 4.5 to pH = 7.2; and - step H): separating the husk-free pulp, the pH of which has been shifted to the acidic range in step G) into a Date Recue/Date Received 2022-01-18 plurality of phases, wherein at least one of said plurality of phases is a polyphenol-albumin liquid phase;
- step I): isolating at least one of the sinapic acid and the salt of sinapic acid from the husk-free pulp of step F) or G) or from the polyphenol-albumin liquid phase of step H) directly or after the husk-free pulp passes through further intermediate steps, by an extraction with an extraction medium.
2. The method as claimed in claim 1, wherein the native material mixture comprises complete seeds or previously partially de-oiled seeds.
3. The method as claimed in claim 2, wherein the previously partially de-oiled seeds comprises a press cake which remains as a residue when oil is pressed off using a press.
4. The method as claimed in any one of claims 1 to 3, wherein comminuting the husks comprises breaking open the husks.
5. The method as claimed in any one of claims 1 to 4, wherein up to a maximum of 6 parts of water are added in step C).
6. The method as claimed in any one of claims 1 to 5, wherein up to a maximum of 5 parts of water are added in step C).
7. The method as claimed in any one of claims 1 to 6, wherein the water-soluble organic solvent is ethanol.
8.
The method as claimed in claim 7, wherein the ethanol is in water-diluted form and is added in such a manner that an alcohol concentration is achieved in step E) which is less than 30%.
Date Recue/Date Received 2022-01-18 9.
The method as claimed in any one of claims 1 to 6, wherein the water-soluble organic solvent is a linear aliphatic alcohol.
10. The method as claimed in any one of claims 1 to 9, wherein step F) is performed in a centrifuge in a centrifugal field.
11. The method as claimed in any one of claims 1 to 9, wherein step H) is performed in a centrifuge comprising at least one of a decanter and a separator.
12. The method as claimed in any one of claims 1 to 11, characterized in that the separating into a plurality of phases of step H) is performed in at least one step to produce at least one of an oil-containing phase having a triglycerol content and an aqueous phase having albumin and sinapic acid content.
13. The method as claimed in any one of claims 1 to 11, characterized in that the separating into a plurality of phases of step H) is performed in at least one step resulting in at least one aqueous phase having an albumin content and sinapic acid content and residual oil content.
14. The method as claimed in any one of claims 1 to 13, wherein the extraction medium used is ethyl acetate.
15. The method as claimed in any one of claims 1 to 13, wherein the extraction medium used is pentanol.
16. The method as claimed in any one of claims 1 to 15, wherein the isolating by extraction step of step I) comprises a multistage extraction process.
Date Recue/Date Received 2022-01-18 17. The method as claimed in any one of claims 1 to 16, wherein the extraction medium is vaporized.
18. The method as claimed in any one of claims 1 to 17, wherein the native material mixture is cold-pressed in a preliminary stage, and undergoes step A) no more than 31 days later.
19. The method as claimed in claim 18, wherein the native material mixture undergoes step A) no more than one of 3 days,48 hours, and 24 hours later.
20. The method as claimed in any one of claims 1 to 17, wherein the native material mixture used in step A) is a cold-pressed material comprising a press cake.
21. The method as claimed in claim 20 in which the press cake is an oil seed rape press cake which has been pressed at a temperature below 70 C.
22. The method as claimed in claim 21 in which the press cake is pressed at a temperature below 60 C.
23. The method as claimed in any one of claims 1 to 22, wherein at least one of the separation steps F) and H) occurs in one of a three-phase decanter and in two-phase decanters in at least two steps.
24. The method as claimed in any one of claims 1 to 22, wherein at least one of the separation steps F) and H) occurs in a nozzle separator.
25. The method as claimed in any one of claims 1 to 24, wherein in step E)the content of the water-soluble organic solvent in an Date Recue/Date Received 2022-01-18 aqueous fraction of the pulp after the addition of the water-soluble organic solvent is less than 45% by volume.
26. The method as claimed in claim 25 wherein the content of the water-soluble organic solvent in an aqueous fraction of the pulp is less than 30% by volume.
27. The method as claimed in claim 25 or 26 wherein the content of the water-soluble organic solvent in an aqueous fraction of the pulp is less than 15% by volume.
28. The method as claimed in any one of claims 1 to 27, characterized in that the temperature during method steps A) to I), is below 60 C.
Date Recue/Date Received 2022-01-18
CLAIMED ARE DEFINED AS FOLLOWS:
1.
A method for obtaining at least one of sinapic acid and a salt of sinapic acid from a native material mixture comprising the following steps:
- step A): providing a native material mixture of seeds/fruits having hard frangible husks comprising oil seed rape fruits of the Brassicaceae family;
- step B): if the native material mixture of step A is not yet comminuted: comminuting the native material mixture;
- step C): dispersing the comminuted native material mixture of step A) or B) with water, wherein, to one part of comminuted native material mixture up to a maximum of 8 parts of water are added, and wherein the water and the comminuted native material mixture are agitated such that one of a free-flowing pulp and a dispersion results;
- step D): setting the pH of the pulp from step C) to an alkaline range pH > 9.5;
- step E): adding a water-soluble organic solvent, to the pulp from step C) subsequent to the setting of the pH of the pulp in step D) in order to detach the husks from endosperm of the seeds and fruits;
- step F): separating off a solids phase which has the predominant fraction of the husks;
- step G): shifting the pH of the pulp which is freed from husks from step F) to the pH range from pH = 4.5 to pH = 7.2; and - step H): separating the husk-free pulp, the pH of which has been shifted to the acidic range in step G) into a Date Recue/Date Received 2022-01-18 plurality of phases, wherein at least one of said plurality of phases is a polyphenol-albumin liquid phase;
- step I): isolating at least one of the sinapic acid and the salt of sinapic acid from the husk-free pulp of step F) or G) or from the polyphenol-albumin liquid phase of step H) directly or after the husk-free pulp passes through further intermediate steps, by an extraction with an extraction medium.
2. The method as claimed in claim 1, wherein the native material mixture comprises complete seeds or previously partially de-oiled seeds.
3. The method as claimed in claim 2, wherein the previously partially de-oiled seeds comprises a press cake which remains as a residue when oil is pressed off using a press.
4. The method as claimed in any one of claims 1 to 3, wherein comminuting the husks comprises breaking open the husks.
5. The method as claimed in any one of claims 1 to 4, wherein up to a maximum of 6 parts of water are added in step C).
6. The method as claimed in any one of claims 1 to 5, wherein up to a maximum of 5 parts of water are added in step C).
7. The method as claimed in any one of claims 1 to 6, wherein the water-soluble organic solvent is ethanol.
8.
The method as claimed in claim 7, wherein the ethanol is in water-diluted form and is added in such a manner that an alcohol concentration is achieved in step E) which is less than 30%.
Date Recue/Date Received 2022-01-18 9.
The method as claimed in any one of claims 1 to 6, wherein the water-soluble organic solvent is a linear aliphatic alcohol.
10. The method as claimed in any one of claims 1 to 9, wherein step F) is performed in a centrifuge in a centrifugal field.
11. The method as claimed in any one of claims 1 to 9, wherein step H) is performed in a centrifuge comprising at least one of a decanter and a separator.
12. The method as claimed in any one of claims 1 to 11, characterized in that the separating into a plurality of phases of step H) is performed in at least one step to produce at least one of an oil-containing phase having a triglycerol content and an aqueous phase having albumin and sinapic acid content.
13. The method as claimed in any one of claims 1 to 11, characterized in that the separating into a plurality of phases of step H) is performed in at least one step resulting in at least one aqueous phase having an albumin content and sinapic acid content and residual oil content.
14. The method as claimed in any one of claims 1 to 13, wherein the extraction medium used is ethyl acetate.
15. The method as claimed in any one of claims 1 to 13, wherein the extraction medium used is pentanol.
16. The method as claimed in any one of claims 1 to 15, wherein the isolating by extraction step of step I) comprises a multistage extraction process.
Date Recue/Date Received 2022-01-18 17. The method as claimed in any one of claims 1 to 16, wherein the extraction medium is vaporized.
18. The method as claimed in any one of claims 1 to 17, wherein the native material mixture is cold-pressed in a preliminary stage, and undergoes step A) no more than 31 days later.
19. The method as claimed in claim 18, wherein the native material mixture undergoes step A) no more than one of 3 days,48 hours, and 24 hours later.
20. The method as claimed in any one of claims 1 to 17, wherein the native material mixture used in step A) is a cold-pressed material comprising a press cake.
21. The method as claimed in claim 20 in which the press cake is an oil seed rape press cake which has been pressed at a temperature below 70 C.
22. The method as claimed in claim 21 in which the press cake is pressed at a temperature below 60 C.
23. The method as claimed in any one of claims 1 to 22, wherein at least one of the separation steps F) and H) occurs in one of a three-phase decanter and in two-phase decanters in at least two steps.
24. The method as claimed in any one of claims 1 to 22, wherein at least one of the separation steps F) and H) occurs in a nozzle separator.
25. The method as claimed in any one of claims 1 to 24, wherein in step E)the content of the water-soluble organic solvent in an Date Recue/Date Received 2022-01-18 aqueous fraction of the pulp after the addition of the water-soluble organic solvent is less than 45% by volume.
26. The method as claimed in claim 25 wherein the content of the water-soluble organic solvent in an aqueous fraction of the pulp is less than 30% by volume.
27. The method as claimed in claim 25 or 26 wherein the content of the water-soluble organic solvent in an aqueous fraction of the pulp is less than 15% by volume.
28. The method as claimed in any one of claims 1 to 27, characterized in that the temperature during method steps A) to I), is below 60 C.
Date Recue/Date Received 2022-01-18
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE102014107607.9 | 2014-05-28 | ||
| DE102014107607.9A DE102014107607A1 (en) | 2014-05-28 | 2014-05-28 | Process for recovering sinapinic acid from a native composition |
| PCT/EP2015/061644 WO2015181203A1 (en) | 2014-05-28 | 2015-05-27 | Method for obtaining sinapic acid from a native material mixture |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2948964A1 CA2948964A1 (en) | 2015-12-03 |
| CA2948964C true CA2948964C (en) | 2022-10-18 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2948964A Active CA2948964C (en) | 2014-05-28 | 2015-05-27 | Method for obtaining sinapic acid from a native material mixture |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP3154375B1 (en) |
| CA (1) | CA2948964C (en) |
| DE (1) | DE102014107607A1 (en) |
| PL (1) | PL3154375T3 (en) |
| WO (1) | WO2015181203A1 (en) |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE102016115911B4 (en) * | 2016-08-26 | 2020-07-16 | Gea Mechanical Equipment Gmbh | Process for obtaining a product of value and product of value |
| FR3131679A1 (en) | 2022-01-13 | 2023-07-14 | Institut national des sciences et industries du vivant et de l'environnement - AgroParisTech | SEMI-INTEGRATED PROCESS FOR ENZYME-ASSISTED EXTRACTION AND PURIFICATION OF P-HYDROXYCINNAMIC ACIDS |
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|---|---|---|---|---|
| CA2398053C (en) | 2000-04-12 | 2011-02-01 | Westfalia Separator Industry Gmbh | Method for the fractionation of oil and polar lipid-containing native raw materials |
| DE102007022662A1 (en) * | 2007-05-15 | 2008-11-20 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Phenol extracts from oilseeds |
| DE102011050905A1 (en) | 2011-06-07 | 2012-12-13 | Gea Mechanical Equipment Gmbh | Process for recovering proteins from a native composition |
| CA2893703C (en) * | 2012-12-27 | 2021-03-16 | Gea Mechanical Equipment Gmbh | Method for obtaining valuable products, in particular proteins, from a native mixture of materials |
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2014
- 2014-05-28 DE DE102014107607.9A patent/DE102014107607A1/en not_active Withdrawn
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2015
- 2015-05-27 CA CA2948964A patent/CA2948964C/en active Active
- 2015-05-27 PL PL15726571T patent/PL3154375T3/en unknown
- 2015-05-27 WO PCT/EP2015/061644 patent/WO2015181203A1/en active Application Filing
- 2015-05-27 EP EP15726571.1A patent/EP3154375B1/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| WO2015181203A1 (en) | 2015-12-03 |
| EP3154375A1 (en) | 2017-04-19 |
| CA2948964A1 (en) | 2015-12-03 |
| DE102014107607A1 (en) | 2015-12-03 |
| EP3154375B1 (en) | 2018-07-18 |
| PL3154375T3 (en) | 2018-12-31 |
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