Method for extracting cocoa and extract obtained by this method

Abstract

A method for obtaining an extract from cocoa fruit components is described, comprising the steps of: a) subjecting cocoa fruit components to wet grinding in a polar solvent and separating the liquid phase from the wet grinding mixture to obtain a hydrophilic extract; b) optionally separating residual solids from the hydrophilic extract; c) adsorbing the hydrophilic extract onto a nonionic macroporous resin; d) desorbing the hydrophilic extract from the nonionic macroporous resin using a water-miscible organic solvent to obtain a polyphenolic extract; and e) concentrating the polyphenolic extract to obtain a polyphenolic concentrate. Further embodiments describe the extract obtained by the above method, and food compositions, supplements, medical compositions, or cosmetic compositions containing the above extract.

Description

Technical Field

[0001] The present invention relates to an improved method for obtaining extracts of polyphenols, theobromine enrichment, and protein enrichment from cacao fruit constituents using phase separation and adsorption / desorption techniques. In certain embodiments, the invention relates to extracts obtained by the above method and compositions containing the same.

Background Art

[0002] Cacao (Theobroma cacao L.) is widely recognized as an important source of health components such as minerals, vitamins, and especially polyphenols (e.g., catechins, flavanol glycosides, anthocyanins, and procyanidins). The main polyphenol compounds present in cacao beans are monomeric flavonoids such as (+)-catechin and (−)-epicatechin, and oligomeric flavonoids such as proanthocyanidins, which reduce the risk of cardiovascular events, bring about blood pressure lowering activity, antiplatelet, anti-inflammatory, metabolic, and anti-atherosclerotic effects, and also improve endothelial function (see A.C. Aprotosoaie et al., Diseases 2016, 4(4), 39).

[0003] Since cacao beans are one of the most well-known sources of dietary polyphenols, numerous extraction methods have been developed for obtaining polyphenol-rich extracts from cacao beans.

[0004] An exemplary method for extracting cacao polyphenols from cacao beans is disclosed in US8451045B2. US6159451A discloses a method for preparing an extract of the shell of cacao beans having improved activity against glucosyltransferase in the prevention of dental caries.

[0005] Generally, it is desirable to provide highly concentrated polyphenolic extracts in powder form having the highest possible purity.

[0006] In this regard, EP2071961B1 discloses a method for obtaining a polyphenol-rich cocoa powder extract, characterized by subjecting freshly depulped cocoa seeds to a degreasing process by blanching, drying, and pressing, stabilizing the degreasing product, grinding it, and then obtaining it through subsequent solid-liquid extraction, liquid-liquid extraction, and concentration steps.

[0007] WO2005 / 115160A1 and EP1748700A1 disclose a method for obtaining a cocoa polyphenol concentrate by subjecting unfermented cocoa beans to a blanching process, drying the unfermented cocoa beans, subjecting the dried unfermented cocoa beans to a particle size reduction process, extracting polyphenols from the dried unfermented cocoa bean intermediate to obtain a cocoa polyphenol extract and extracted solids, and concentrating the cocoa polyphenol extract to obtain a cocoa polyphenol concentrate. This method further includes a degreasing step performed by pressing the cocoa bean raw material before the extraction step.

[0008] However, the total polyphenol yield of these methods, based on the initial polyphenol content in the starting materials, has room for improvement. For example, EP2071961B1 discloses a total polyphenol yield of 11.93%.

[0009] Furthermore, it is not easy to provide polyphenol extracts that are not only highly concentrated but also ensure desirable bioavailability. Monomeric flavanols have been shown to be better absorbed from the intestines than polymerized proanthocyanidins and therefore exhibit higher antioxidant activity in vivo. However, the liquid-liquid extraction process can result in an oligomer distribution favorable to higher-order oligomers with a degree of polymerization (DP) of 3 or higher (see, e.g., EP1913821A1). In addition, numerous processing steps, particularly those involving the input of thermal and / or mechanical energy, can lead to the degradation, epimerization, and oxidative polymerization of polyphenols (see, e.g., J. Alean et al., Journal of Food Engineering 2016, 189, 99-105).

[0010] To avoid the use of mechanical pressing for the separation of fat (i.e., cocoa butter) and to minimize the heat load on the cocoa bean raw material, alternative cocoa bean processing methods have been developed to preserve a higher content of valuable heat-sensitive components.

[0011] For example, WO2010 / 073117A1, EP3114940A1, EP3114941A1, EP3114942A1 and EP3114939A1 disclose methods for processing cocoa beans, which include forming a suspension containing cocoa beans or nibs and water, wet grinding of the suspended beans or nibs, heating of the suspension, and decanting the suspension so that it separates into an aqueous phase, a fatty phase, and a solid phase, which results in a lower heat load on the cocoa mass compared to conventional methods, thus preserving a higher content of polyphenols, antioxidants, and / or vitamins.

[0012] Advantageously, these methods assume the production of chocolate products using a chocolate building kit, which includes cocoa butter, cocoa powder, cocoa flavoring, and polyphenolic powder extracts, allowing polyphenol extraction to be fully integrated into the chocolate manufacturing process. Here, the polyphenolic powder is obtained by subjecting the aqueous phase to a dearomatization step, a concentration step (generally evaporation of excess water), and optionally spray drying. However, these methods have been found unsuitable for producing highly concentrated polyphenolic powder with a total flavanol content higher than 10 g per 100 g of defatted dry material, as the content generally ranges from 2 to 8 g per 100 g of defatted dry material, depending on the cultivar.

[0013] Considering the above, it remains desirable to provide a method for producing a polyphenol extract that yields an improved total polyphenol content relative to the total polyphenol content in the cacao fruit components, and simultaneously has a high concentration of flavanols and a high ratio of monomeric and dimeric flavanols.

[0014] In addition to polyphenols, cocoa and cocoa products also contain large amounts of methylxanthines, namely theobromine (known to be a brain stimulant, diuretic, and potentially blood pressure-lowering substance) and caffeine, which account for approximately 2.4–3.6% of the defatted dried cocoa composition. Commonly used methods for extracting theobromine from cocoa beans often involve extraction with chloroalkanes (e.g., chloroform or dichloromethane), as shown, for example, in US1,925,326A. However, the use of such extraction solvents is not recommended from an environmental standpoint, as well as from the fact that the residue of such solvents in food is undesirable and requires extensive removal processes. [Overview of the project] [Problems that the invention aims to solve]

[0015] Therefore, in the value chain of cocoa bean processing for use in the preparation of food compositions, supplements, medical compositions, or cosmetic compositions, it is desirable to efficiently extract the above-mentioned constituent substances and simultaneously provide further extracts. [Means for solving the problem]

[0016] The present invention solves this objective using the subject matter of the claims as defined herein. The advantages of the present invention will be described in more detail in the following sections, and further advantages will become apparent to those skilled in the art when considering the disclosure of the invention.

[0017] In one embodiment, the present invention relates to a method for obtaining an extract from cocoa fruit components, comprising the steps of: a) subjecting cocoa fruit components to wet grinding in a polar solvent and separating the liquid phase from the wet grinding mixture to obtain a hydrophilic extract; b) optionally separating residual solids from the hydrophilic extract; c) adsorbing the hydrophilic extract onto a nonionic macroporous resin; d) desorbing the hydrophilic extract from the nonionic macroporous resin using a water-miscible organic solvent to obtain a polyphenolic extract; and e) concentrating the polyphenolic extract to obtain a polyphenolic concentrate.

[0018] In another aspect, the present invention relates to a polyphenolic extract obtained by the above-described method, which preferably has the following characteristics: a total flavanol content determined by HPLC, preferably at least 25 g per 100 g of defatted dry material, more preferably at least 30 g per 100 g of defatted dry material, even more preferably at least 45 g per 100 g of defatted dry material, even more preferably 50 g per 100 g of defatted dry material, particularly preferably at least 53 g per 100 g of defatted dry material, and most preferably at least 75 g per 100 g of defatted dry material; a degree of polymerization of 1 to 7 ( The product satisfies at least one of the following conditions: a content of monomeric flavanols (DP1) determined by HPLC, at least 18% by weight, preferably at least 20% by weight, and more preferably at least 24% by weight, relative to the total content of flavanols having DP1 to DP7; a protein content of less than 42 g per 100 g of defatted dry material; a carbohydrate content of less than 6 g per 100 g of defatted dry material; and a dietary fiber content of less than 2 g per 100 g of defatted dry material, where the protein, carbohydrate, and dietary fiber content can be determined by methods known in the art.

[0019] In a further embodiment, the present invention relates to a protein / dietary fiber concentrated extract obtained by the method described above, in which case the residual solids are further treated by at least a concentration step and preferably by spray drying.

[0020] In yet another aspect, the present invention relates to a protein-enriched extract obtained by the method described above, in which case the non-adsorbed fraction of the hydrophilic extract in step c) is further processed by at least a concentration step.

[0021] In a further aspect, the present invention relates to a food composition, supplement, medical composition or cosmetic composition comprising the extract described above. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] [Figure 1] It is a diagram for explaining an exemplary method for obtaining an extract from cacao fruit constituents according to the present invention. [Figure 2] It is a diagram schematically showing an exemplary method for producing chocolate using the extract provided by the present invention. MODE FOR CARRYING OUT THE INVENTION

[0023] Extraction and fractionation method In a first embodiment, the present invention is a method for obtaining an extract from cacao fruit constituents, comprising: a) subjecting the cacao fruit constituents to wet grinding in a polar solvent and separating a liquid phase from the wet grinding mixture to obtain a hydrophilic extract; b) optionally, separating residual solids from the hydrophilic extract; c) adsorbing the hydrophilic extract onto a non-ionic macroporous resin; d) desorbing the hydrophilic extract from the non-ionic macroporous resin using a water-miscible organic solvent to obtain a polyphenolic extract; and e) concentrating the polyphenolic extract to obtain a polyphenolic concentrate.

[0024] Surprisingly, the series of processing steps a) to e) were found to result in an improved yield of total polyphenols relative to the total polyphenol content in the cocoa fruit components, and to produce a polyphenol extract that simultaneously has a high concentration of flavanols and a high ratio of monomeric and dimeric flavanols. Furthermore, since this method does not require the use of organic solvents that are unsuitable for food (e.g., hexane or haloalkanes, e.g., chloroform, dichloromethane or tetrachloromethane), it can be directly incorporated into the preparation of food-safe cocoa products (e.g., chocolate, cocoa beverages, cocoa-based desserts, supplements or similar) while minimizing the number of processing steps required.

[0025] As used herein, the term “cacao fruit components” refers to one or more components of a Theobroma cacao fruit pod, which generally consists of about 30 to 50 fruit seeds (i.e., cacao beans) enclosed in a cacao pod shell surrounded by pulp and mucus. Thus, such cacao fruit components may include cacao beans, cacao bean husk (CBS) material, cacao pulp / mucus and / or cacao pod shell material. In a preferred embodiment of the present invention, the cacao fruit components subjected to step a) include cacao beans, which may also include cacao nibs.

[0026] Non-limiting examples of the method of the present invention are illustrated in the flowchart shown in Figure 1.

[0027] For the purposes of the present invention, the cocoa beans or nibs may be fermented or unfermented, dried or undried, roasted or unroasted, and / or depulped or undepulped.

[0028] However, the total polyphenol content of defatted dried cocoa beans is generally about 12-20%, and about 5% in fermented beans. Generally, the polyphenol content in cocoa beans depends on their origin and processing (fermentation, drying, alkalization, roasting, etc.), which can substantially affect the polyphenol content and consequently affect the quality of the final product. Since up to 70-80% of polyphenols can be broken down during the fermentation process (see, for example, US2004 / 0096566A1), the cocoa fruit components subjected to step a) preferably include or consist of unfermented cocoa beans. The term "fermentation" generally refers to any activity or process involving the enzymatic or metabolic breakdown (digestion) of organic matter by microorganisms, and includes both anaerobic and aerobic processes, as well as processes involving a combination or sequence of one or more anaerobic and / or aerobic stages. As used herein, the term "unfermented" refers to the degree of fermentation of cocoa beans. As is known to those skilled in the art, the degree of fermentation is indicated by the fermentation index (see Shamsuddin and Dimick, "Proceedings of the Cacao Biotechnology Symposium," edited by PSDimick, The Pennsylvania State University, 1986, Qualitative and Quantitative Measurements of Cacao Bean Fermentation, pp. 55–74). Generally, the fermentation index of unfermented beans is less than 0.5. It may be even more preferable to subject the cacao beans to step a) immediately after opening the cacao fruit pods, separating the beans from the pod shells, and optionally removing the pulp / mucus. Further examples of preferred cacao beans include lavado beans, which are unfermented beans (generally terracotta in color) made by taking the cacao beans from ripe pods, washing the beans with water to remove the pulp, and drying them (including, but not limited to, sun-drying).

[0029] Furthermore, polyphenols are generally known to be heat-sensitive components. Therefore, in order to preserve as much of the original polyphenol content of the cocoa beans as possible, the cocoa fruit components subjected to step a) preferably include cocoa beans that have not undergone the drying and / or roasting process.

[0030] In a further preferred embodiment, unfermented cocoa beans may be pretreated in an incubation step, with or without cocoa pulp and mucus, prior to step (a). Such an incubation step is characterized by inhibiting germination of the cocoa beans by specific physical and / or chemical pretreatment immediately after the beans are removed from the pod (with or without pulp and mucus).

[0031] In particular, sprouting of freshly harvested cocoa beans can be suppressed by incubating them in an incubation medium at a high temperature, for example, between 10°C and 70°C, preferably between 10°C and 55°C, for a period between 2 hours and 10 days, preferably between 3 hours and 168 hours. As is known to those skilled in the art, in order to suppress naturally occurring fermentation as effectively as possible, it is especially preferable to transfer the cocoa beans (optionally together with pulp / mucus) to the incubation medium immediately after opening the pod, preferably under sterile conditions. The incubation medium is not particularly limited and may be, for example, an aqueous acidic medium used in the cocoa bean processing method disclosed in US8,501256B2, or, for example, an aqueous ethanol solution.

[0032] Furthermore, polyphenols are generally known to be heat-sensitive components. Therefore, in order to preserve as much of the original polyphenol content of the cocoa beans as possible, the cocoa fruit components subjected to step a) preferably include cocoa beans that have not undergone a drying and / or roasting process, which involves heat treatment at over 100°C or preferably over 70°C.

[0033] In step a), the cacao fruit components are subjected to wet grinding in a polar solvent, and the liquid phase is separated from the wet-ground mixture to obtain a hydrophilic extract.

[0034] The polar solvents used herein generally have a dielectric constant of 15 or more, preferably 50 or more, and more preferably 80 or more. The polar solvent may include polar protic solvents and polar aprotic solvents, but polar protic solvents are preferably used. In preferred embodiments, the polar solvent is water, C1-C8 alcohols (e.g., methanol, ethanol, isopropanol), C2-C8 ketones (e.g., acetone), C3-C7 esters (e.g., methyl acetate, ethyl acetate), C2-C8 ethers (e.g., diethyl ether, methyl t-butyl ether), C4-C 10 The solvent is selected from lactates and mixtures thereof. Water or an ethanolic water mixture is particularly preferred. It is even more preferable that the polar solvent does not contain halogenated hydrocarbons.

[0035] In a preferred embodiment, the polar solvent contains or consists of water. In another preferred embodiment, the polar solvent may be mixed with one or more organic solvents, which may or may not be miscible with water under extraction conditions, to contain water in a content of less than 95% by weight (e.g., less than 90%, less than 80%, less than 50%, less than 30%, less than 10%, or less than 5%). Naturally, alternative aqueous liquids may be used as a water source, preferably selected from one or more liquids having a water content of 60 to about 95% by weight, such as coffee, tea, and fruit juices, fruit juice concentrates, or milk.

[0036] In preferred embodiments, the polar solvent may also contain an acidic component (but not limited to acetic acid, citric acid, or other acids) in addition to acetic acid and / or citric acid formed by microbial activity in the beans, which can promote efficient extraction at low temperatures and / or reduce the oxidative degradation of polyphenols.

[0037] According to the present invention, in order to avoid or reduce the destruction of cellular compartments in cocoa raw materials caused by high mechanical load or shear stress associated with dry grinding methods conventionally used in the art, the cocoa bean components are wet-ground in the presence of a polar solvent.

[0038] While not specifically limited, the weight ratio of the polar solvent to the cocoa bean components in the suspension subjected to wet grinding is preferably between 1:1 and 6:1, more preferably between 2:1 and 4:1, and particularly preferably about 3:1, and these ratios may favorably affect processability in further steps (e.g., facilitated pumping, grinding, and easier phase separation).

[0039] It is preferable to wet-grind the cocoa fruit components using one or more wet-grinding steps to an average particle size of 100 μm or less, preferably 50 μm or less, and more preferably 20 μm or less. Grinding the cocoa fruit components to this dimensional range substantially increases the exposed surface area of ​​the granular material, thereby enabling more efficient wetting for improved extraction of polyphenols. Particle size reduction can be achieved, for example, by using a disc mill (e.g., a perforated disc mill), a colloid mill (e.g., a toothed colloid mill), or a corundum mill.

[0040] In at least one grinding step, it is preferable to liquefy the cocoa fruit components (e.g., cocoa beans or nibs) to increase the available surface area of ​​the liquefied solids, thereby allowing the polar solvent to better wet the solids. The methods and apparatus used for wet grinding are not particularly limited, as long as significant frictional heat generation or undesirable emulsification due to large mechanical forces is avoided. For example, when multiple grinding steps are used, a coarse wet grinding step (e.g., optionally using an additional polar solvent) may be performed using a perforated disc mill, and the coarse grinding suspension may be pumped to a toothed colloid mill for the fine grinding step. When multiple grinding steps are used, it is understood that the polar solvent may be modified or altered (e.g., by adding another solvent or acid). In a preferred embodiment, liquefaction occurs by cold grinding (one or more steps) at a temperature of 2 to 25°C.

[0041] In a preferred embodiment of the present invention, step a) includes a1) adding a polar solvent to cocoa fruit components to form a suspension, a2) wet grinding the suspension, a3) heat treating the suspension at a temperature of 70°C or lower, and a4) separating the suspension into a hydrophilic liquid phase (heavy phase), a hydrophobic liquid phase (light phase), and a solid phase, wherein the hydrophilic liquid phase contains a hydrophilic extract as a major component and residual solids as a trace component, the hydrophobic liquid phase contains cocoa butter as a major component and solids and / or a hydrophilic solvent as a trace component, and the solid phase contains cocoa powder and a hydrophilic solvent. Steps a1) and a2) may be carried out based on the preparation and grinding of the suspension according to the above. In step a3), in order to reduce the overall heat load and prevent emulsification of the cocoa bean raw material, the wet-ground suspension is heat treated at a temperature of about 70°C or lower. From the viewpoint of a favorable balance between cocoa butter yield and the preservation of desirable flavors such as aromatic substances, antioxidants, and / or vitamins, a heating temperature of 43 to 65°C is preferred. From the viewpoint of liquefaction of cocoa butter and / or improved mechanical phase separation, a heating temperature range of 45 to 50°C is particularly preferred. Heating of the wet-ground suspension can be carried out by a scraping or tubular heat exchanger, but is not limited to these.

[0042] Subsequently, in step a4), the suspension is separated into a hydrophilic liquid phase (heavy phase), a hydrophobic liquid phase (light phase), and a solid phase. The hydrophilic liquid phase contains a hydrophilic extract as its main component and residual solids as a trace component; the hydrophobic liquid phase contains cocoa butter as its main component and solids and / or a hydrophilic solvent as a trace component; and the solid phase contains cocoa powder and a hydrophilic solvent. Furthermore, the solid phase may contain residual cocoa butter in an amount up to 30% by weight, preferably less than 27% by weight, and more preferably less than 20% by weight, relative to the total dry weight.

[0043] Preferably, mechanical particle separation can be achieved using a device that utilizes centrifugal force, such as a decanter or nozzle separator. For example, the suspension may be decanted to separate coarse or large or high-mass solids from the liquid, and then smaller and / or finer solid particles may be further separated from the liquid, and / or oil products may be separated from non-oil products.

[0044] Multiple phase separation and recombination steps can be used to achieve improved separation between the hydrophilic liquid phase (heavy phase), the hydrophobic liquid phase (light phase), and the solid phase. For example, the hydrophobic liquid phase obtained by the initial decanting step may be further filtered or centrifuged to separate any remaining particulate matter or water from the hydrophobic liquid phase, and the resulting particulate matter and polar solvent may be recombined with the polar solvent and solid phase from the initial decanting step or from a subsequent processing step of the phase.

[0045] Once the three phases are separated, they may be processed independently to separate, for example, cocoa butter (from the hydrophobic liquid phase), cocoa powder (from the solid phase), and cocoa flavor (from any of the three phases). Examples of independent processing of the phases and their recombination are disclosed in WO2010 / 073117A1, EP3114940A1, EP3114941A1, EP3114942A1, EP3114939A1, EP3747277A1, EP3747275A1, and EP3747276A1.

[0046] According to the present invention, the hydrophilic liquid phase obtained in step a) is further processed to extract a polyphenolic concentrate.

[0047] If acids were added during step a) at this stage, they may optionally be removed from the hydrophilic liquid phase or neutralized. The appropriate removal method is not particularly limited and can be carried out by any suitable method known in the art (e.g., by extractive distillation, reactive distillation, extractive (e.g., liquid-liquid) extraction, emulsifying liquid film treatment, salting out, or a combination thereof).

[0048] Generally, the hydrophilic liquid phase obtained in step a) (or step a4)) respectively contains the remaining solids and cocoa butter. According to the present invention, it is preferable to separate the residual solids from the hydrophilic extract in step c) in order to remove particulate matter and cocoa butter residue that may adversely affect the adsorption process. For this purpose, it is preferable to subject the hydrophilic liquid phase to the filtration step in step b). In a further preferred embodiment, the filtration process and clarification can be enhanced by pre-adding filter aids and / or protein precipitants. Examples of filter aids include, but are not limited to, cellulose and cellulose derivatives. Furthermore, the pH of the hydrophilic liquid phase can be adjusted by adding acids and / or bases up to the isoelectric point to achieve isoelectric precipitation and promote the efficiency of subsequent filtration. Examples of protein precipitants include carrageenan, agar, alginic acid, bentonite, pectin, colloidal silica, tannins, or combinations thereof.

[0049] In particularly preferred embodiments, cross-flow filtration and / or filtration through a membrane filter having an absolute pore size rating of 0.45 μm or less, such as 0.02 to 0.45, is used. Dynamic cross-flow filtration using a ceramic membrane is particularly preferred, in which case the cross-flow effect (= tangential overflow of the membrane surface) is achieved by rotating the filter disc and not pumping a large amount of liquid. Therefore, efficient filtration can be performed continuously without clogging of the filter even with relatively high solid loads. It is even more preferable to use a membrane filter having an absolute pore size rating of 0.30 μm or less, such as 0.25 or less, 0.22 μm or less, 0.20 μm or less, or 0.10 μm or less. In some embodiments, the use of a membrane filter having an absolute pore size rating of 0.20 μm or less may be particularly preferred. For example, filtration using a membrane filter with a pore size rating between 0.04 μm and 0.16 μm, more preferably between 0.06 μm and 0.10 μm, has been shown to favorably enable higher flavonoid concentrations in the desorbed polyphenolic extract without substantially reducing the flavonoid yield.

[0050] In preferred embodiments, filtration may be performed at a temperature of 3 to 50°C, for example, between 25°C and 35°C. However, filtration at a temperature between 4 to 15°C, or between 5°C and 10°C, is particularly preferred.

[0051] In a particularly preferred embodiment, a dialysis filtration step is further incorporated to facilitate flavonoid extraction by introducing a purified solvent (e.g., nanofiltered (NF) water or reverse osmosis (RO) water) into the process fluid. Preferably, the volume of the purified solvent is adjusted to 45% or less of the total volume of the feed product (each cocoa extract).

[0052] The retained liquid (i.e., residual solids) obtained during filtration can be further processed by at least a concentration step to obtain a protein concentrate extract, which may occur by evaporation of residual polar solvents in one or more steps. The protein concentrate extract generally contains a large proportion of dietary fiber and can therefore also be understood as a protein / dietary fiber concentrate extract (P / DF concentrate extract). Optionally, the protein concentrate extract thus obtained can be subjected to spray drying to obtain a cocoa-based protein concentrate powder or protein / dietary fiber concentrate powder (P / DF concentrate powder), which represents additional components that are valuable but were previously frequently discarded.

[0053] The permeate obtained in the filtration process (i.e., the clear hydrophilic extract) generally contains less than 1 g of cocoa butter per 100 g of dry material. The main components of flavanols and theobromine contained in the hydrophilic phase are generally still present in the permeate along with a large amount of amino acids.

[0054] In step c), the hydrophilic extract thus obtained may be optionally and gently heated (below 70°C, preferably below 60°C) to adsorb it onto a nonionic macroporous resin. For this purpose, a column containing a nonionic macroporous resin is prepared, washed, and appropriately conditioned by a method known to those skilled in the art. When the hydrophilic extract is pumped through the column, the cocoa polyphenols are adsorbed onto the resin.

[0055] In preferred embodiments, the nonionic macroporous resin comprises or consists of a crosslinked aliphatic polymer, an aliphatic acrylic polymer, or a polyacrylic polymer.

[0056] From the viewpoint of improved bonding ability and process efficiency, nonionic macroporous resins are preferably at least 400 m 2 / g, more preferably at least 450m 2 / g, more preferably at least 500m 2It has a surface area determined by the nitrogen BET method, which is 0.55 cc / g. Alternatively, or in combination, the nonionic macroporous resin preferably has a total pore volume determined by the nitrogen BET method, which is at least 0.55 cc / g, more preferably at least 0.80 cc / g, even more preferably at least 0.85 cc / g, and particularly preferably at least 0.90 cc / g.

[0057] After adsorption, the resin may be optionally washed (e.g., with deionized water) to remove undesirable components. The washing step is preferably carried out at a temperature of 10 to 50°C. In particular, the non-adsorbed fraction of the hydrophilic extract in step c) may be further treated by at least a concentration step to obtain a theobromine concentrated extract.

[0058] In step d), a hydrophilic extract is desorbed from a nonionic macroporous resin by eluting the adsorbate using a water-miscible organic solvent to obtain a polyphenolic extract.

[0059] The water-miscible organic solvent is preferably selected from C1-C8 alcohols, C3-C5 ketones, C3-C5 esters, C2-C4 ethers, C2-C5 aldehydes, and mixtures thereof. Particularly preferred water-miscible organic solvents include methanol, ethanol, isopropanol, and / or acetone. The water-miscible organic solvent may be optionally diluted with water, provided that the water content does not exceed 50% by volume. In a particularly preferred embodiment, an aqueous ethanol solution (e.g., 60-80% by volume ethanol) may be used. It is even more preferable that the water-miscible organic solvent does not contain halogenated hydrocarbons. In a preferred embodiment, an acidic component (not limited to inorganic acids, acetic acid, citric acid, or other acids) may be added to the water-miscible organic solvent.

[0060] In particular, the loaded resin may be eluted sequentially using the same solvent but at different temperatures, or using different solvents. For example, a first solvent suitable for high solubility of caffeine and / or theobromine may be used first to selectively elute alkaloids from a nonionic macroporous resin, and then a second solvent (i.e., a water-miscible organic solvent) may be used to elute polyphenolic components. The desorption process may be appropriately optimized depending on the desired quality of the individual extracts.

[0061] To facilitate the removal of undesirable volatile compounds, the water-miscible organic solvent may be optionally heated before elution.

[0062] In step e), the polyphenol extract obtained in step d) is subjected to a concentration step to obtain a polyphenolic concentrate. Concentration can be achieved by removing the water-miscible solvent in one or more steps, for example by evaporation or distillation, preferably under reduced pressure or vacuum conditions. A series of vacuum distillation / evaporation and spray drying steps are preferred from the viewpoint of stabilizing the polyphenolic components (e.g., flavanols) to obtain a polyphenolic powder.

[0063] In order to preserve as much polyphenol as possible contained in the original cocoa fruit components, the cocoa fruit components and their extracts are preferably not exposed to temperatures above 70°C, at least during steps a) to d), more preferably during steps a) to e), and even more preferably during the entire extraction process.

[0064] Advantageously, the above method can be fully incorporated into methods for preparing cocoa-based products for human consumption (e.g., chocolate, cocoa powder, cocoa butter, etc.) without relying on undesirable solvents, resulting in excellent yields of cocoa polyphenols. Specifically, it is possible to achieve a total polyphenol yield of 15% or more relative to the polyphenol content (100%) in cocoa beans, for example, in the range of 15-30%, and generally at least 20%.

[0065] Cacao extract In a second embodiment, the present invention relates to a polyphenolic extract obtained by the method according to the first embodiment described above.

[0066] Generally, the polyphenolic compounds naturally present in cocoa beans mainly include catechins ((-)-epicatechin, (+)-catechin, (+)-gallocatechin, and (-)epigallocatechin), procyanidins (procyanidins B1-B5, procyanidin C1, procyanidin D, and higher-order oligomers and polymers, mostly homologs of epicatechin having 2-18 monomer units), anthocyanins (i.e., cyanidin-3α-L-arabinoside and cyanidin-3-β-D-galactoside), flavanol glycosides (i.e., quercetin-3-O-α-D-arabinoside and quercetin-3-O-β-D-glucoplanoside), clobamide, and dideoxyclobamide.

[0067] As used herein, the terms "cocoa flavanol" or "flavanol" refer to the monomeric flavanols (-)-epicatechin and (+)-catechin, as well as their associated oligomeric flavanols (procyanidins) having a degree of polymerization (DP) of DP1 to DP7. The individual and total flavanol content based on the degree or polymerization (DP1 to DP7) can be determined using commercially available standards by HPLC coupled with fluorescence detection (e.g., according to U. Bussy et al., Food Funct. 2020, 11(1), 131-138).

[0068] In preferred embodiments, the polyphenolic extract of the present invention exhibits a total flavanol content determined by HPLC of at least 25 g per 100 g of defatted dry material, more preferably at least 30 g per 100 g of defatted dry material, even more preferably at least 45 g per 100 g of defatted dry material, even more preferably at least 50 g per 100 g of defatted dry material, particularly preferably at least 53 g per 100 g of defatted dry material, and most preferably at least 75 g per 100 g of defatted dry material. In particular, the total flavanol content may exceed the total flavanol content of currently available flavanol standards (e.g., NIST Reference Material 8403). The upper limit of the total flavanol content is not particularly limited and may be 100 g per 100 g of defatted dry material.

[0069] Furthermore, or alternatively, the polyphenolic extract of the present invention exhibits a monomeric flavanol (DP1) content, preferably at least 18% by weight, more preferably at least 20% by weight, and even more preferably at least 24% by weight, relative to the total content of flavanols having degrees of polymerization 1 to 7 (DP1 to DP7), as determined by HPLC. It may be even more preferable that the polyphenolic extract of the present invention exhibits a dimeric flavanol (DP2) content, preferably at least 18% by weight, and more preferably at least 20% by weight, relative to the total content of flavanols having degrees of polymerization 1 to 7 (DP1 to DP7), as determined by HPLC. Thus, the polyphenolic extract contains high content of monomeric and dimeric flavanols, which exhibit particularly desirable bioavailability. Furthermore, or alternatively, the polyphenolic extract of the present invention preferably exhibits a protein content of less than 42 g per 100 g of defatted dry material (e.g., 5 to 41 g per 100 g of defatted dry material), a carbohydrate content of less than 6 g per 100 g of defatted dry material (e.g., 2 to 5 g per 100 g of defatted dry material), and a dietary fiber content of less than 2 g per 100 g of defatted dry material (e.g., 0.1 to 1.8 g per 100 g of defatted dry material). The above-mentioned nutrient content is a characteristic feature of the polyphenolic extract prepared by the method of the present invention. The respective contents of protein, carbohydrates, and dietary fiber can be determined by methods for nutrient analysis commonly used in the field, such as the "Big 8" or "Big 12" nutrient analysis methods. The protein content can be determined by titration using the Kjeldahl method, for example, based on the total nitrogen content using a conversion factor of 6.25 (see, for example, AOAC method 970.22). As used herein, the term “dietary fiber” is understood to define indigestible carbohydrates and lignin, and the dietary fiber content may be determined by a method well known to those skilled in the art, for example, by a calculated difference in calorific values ​​determined in accordance with Regulation (EU) No. 1169 / 2011.The (digestible) carbohydrate content can be calculated from the difference between 100% and the total percentage of water, ash, protein, fat, alcohol, and insoluble organic fiber in the sample, in which case the fat content can be determined, for example, by the Weibull-Stoldt method.

[0070] In a third embodiment, the present invention relates to a protein concentrate extract obtained by a method according to the first embodiment. As outlined above, the protein concentrate extract is derived from the retaining liquid (i.e., residual solids) obtained during filtration, which is subjected to a concentration step that achieves the removal of at least polar solvents and further contains cocoa-derived dietary fiber. Optionally, the protein concentrate extract thus obtained may be subjected to spray drying to obtain a cocoa-based protein concentrate powder.

[0071] Generally, protein concentrate powders have a protein content of at least 30% by weight, and usually more than 35% by weight, relative to the total dry weight of protein, carbohydrates, and dietary fiber. The respective contents can be appropriately determined by methods known in the art, such as the Kjeldahl method or the Dumas method, for example, using a conversion factor of 6.25 (see, for example, AOAC method 970.22).

[0072] In a fourth embodiment, the present invention relates to a theobromine concentrate extract obtained by a method according to the first embodiment. As outlined above, the theobromine concentrate extract is derived from the non-adsorbent fraction of the hydrophilic extract in step c), and this non-adsorbent fraction is further treated by at least a concentration step and preferably drying to obtain an extract in powder form. The theobromine concentrate extract generally has a theobromine content of at least 3 g per 100 g of defatted dry material, more preferably at least 4 g per 100 g of defatted dry material, and particularly preferably at least 5 g per 100 g of defatted dry material.

[0073] Composition and use of the extract In the fifth embodiment, the present invention relates to a food composition, supplement, medical composition or cosmetic composition comprising an extract according to the second to fourth embodiments, or a polyphenolic extract, protein concentrate extract and / or theobromine concentrate extract, respectively, prepared according to the first embodiment.

[0074] Naturally, the individual extracts obtained by the method of the present invention may be used and / or sold independently, or incorporated into cocoa bean processing methods for the purpose of providing food and beverage compositions.

[0075] Exemplary food compositions according to the present invention include liquid and solid cocoa-based products suitable for consumption (e.g., chocolate, cocoa-based desserts, cocoa drinks).

[0076] Furthermore, the extract (preferably in powder form) may be used and supplied in the manner of a construction kit for cocoa-based products (e.g., chocolate, cocoa-based desserts, cocoa drinks), which may further include cocoa powder (but not limited to cocoa powder obtained by drying the solid phase obtained in step a) or a4), cocoa butter (but not limited to cocoa butter obtained by processing the hydrophobic liquid phase obtained in step a4), and / or cocoa flavor (but not limited to cocoa flavor recovered by de-aromatherapy of the hydrophilic liquid phase obtained in step a4).

[0077] For example, in the methods of WO2010 / 073117A1, EP3114940A1, EP3114941A1, EP3114942A1, EP3114939A1, EP3747277A1, EP3747275A1 and EP3747276A1, polyphenolic extracts, protein concentrates, and / or theobromine concentrates may be used, preferably in powder form, thereby enabling further fine-tuning of the sensory and / or nutritional properties of the desired product.

[0078] Figure 2 illustrates an example of a method for preparing dark and milk chocolate based on a chocolate construction kit containing cocoa flavoring, cocoa powder, cocoa butter, and polyphenolic extract, protein concentrate extract, and theobromine concentrate extract, manufactured according to the present invention. While not limited to this, the cocoa flavoring may be obtained from de-aromatherapy of the hydrophilic phase, and / or the cocoa flavoring may be a roasted cocoa flavoring obtained from drying / roasting cocoa powder. The kit components are mixed before being subjected to the conching process. Using polyphenolic extracts according to the present invention makes it possible to improve the yield of polyphenols and the bioavailability of the final product compared to prior art processes. Protein concentrate extracts (or protein / dietary fiber concentrate extracts, respectively), which are previously unused fractions in the prior art cocoa bean processing methods described above, may be added or reintroduced to modify the protein content in the final product, thereby ideally and effectively utilizing the cocoa bean components. The flavor may be further adjusted or enhanced by adding one or more sugars, sweeteners, cocoa pulp, and / or fruit juice. Furthermore, theobromine concentrate may be selectively added or reintroduced to utilize the pharmacological effects of alkaloids and / or to balance the bitterness of cocoa-based products.

[0079] For the preparation of milk chocolate, powdered milk is added, preferably before the mixing step. Optionally, emulsifiers (e.g., lecithin) may be added before conching to reduce the viscosity of the chocolate, control the crystallization and flow properties of the sugars, and promote homogeneous mixing of the ingredients. Further ingredients and flavorings, such as vanilla, rum, or similar, may also be added before, during, and after the conching process (e.g., sea salt, fleur de sel, nuts, and raisins). The conching process removes unwanted acetic acid, propionic acid, and butyric acid from the chocolate, reduces moisture, and redistributes flavor-producing substances from the dried cocoa into the fat phase, while maturing the flavor of the product. The conching temperature is controlled and varies depending on the type of chocolate (from approximately 49°C for milk chocolate to 82°C for dark chocolate). While temperature-dependent to some extent, the conching duration in conventional chocolate manufacturing processes generally ranges from 16 hours to a maximum of 72 hours to achieve good results. The conching duration is preferably less than 16 hours, more preferably less than 12 hours, and generally 10 hours or less. Therefore, the loss of desirable aroma characteristics and the decomposition of heat-sensitive components (including polyphenolic fractions) are minimized.

[0080] For the purpose of medical compositions or dietary supplements, extracts according to the present invention can be appropriately administered in forms (e.g., capsules or tablets) and doses appropriately selected by those skilled in the art.

[0081] Due to the antioxidant properties of the polyphenolic components, the polyphenolic concentrate obtained by the present invention can be used in cosmetic applications, such as shampoos, lotions, creams, fragrances, ointments, or similar products.

[0082] Naturally, the features of the first to fifth embodiments may be freely combined in any combination, except for combinations in which at least some of the features are mutually exclusive. [Examples]

[0083] (Example 1) Extraction and phase separation 2569 kg of fermented and debacterized unroasted cocoa nibs (origin: Peru) were mixed with water as a polar solvent in a ratio of approximately 1:3 to form a suspension. The suspension was then wet-milled in multiple stages using a perforated colloidal mill to an average particle size of less than 100 μm and heated to a temperature of approximately 45-60°C using a tubular heat exchanger. Using a three-phase decanter, the two immiscible liquid phases (i.e., hydrophilic and hydrophobic liquid phases) were separated from each other while simultaneously discharging the solid phase. After removing the lighter oil and solid phases, the hydrophilic liquid phase was used to obtain polyphenol extracts, theobromine concentrates, and protein concentrates according to the procedure described in the following sections.

[0084] filtration First, at a temperature of 5-10°C, a ceramic membrane filter disc (commercially available from Novoflow GmbH; filter area, 3 m²) is used. 2 A dynamic cross-flow filtration system with a membrane pore size of 0.2 μm was used to filter the hydrophilic extract. Dialysis filtration was performed by introducing reverse osmosis water at a volume of 10% of the total volume of the hydrophilic extract supply.

[0085] Two products were obtained from the filtration of the aqueous cocoa extract. The retaining liquid mainly contained protein and dietary fiber, while the filtrate mainly contained theobromine in addition to the aqueous solvent, cocoa flavanols, caffeine, and protein. The retaining liquid was then concentrated in an evaporator and converted into protein concentrate powder or protein / dietary fiber concentrate powder (hereinafter abbreviated as P / DF concentrate powder) by spray drying.

[0086] Furthermore, as shown in Table 1, the filtration process effectively removed residual fats (i.e., cocoa butter) from the hydrophilic extract, preventing interference with subsequent adsorption.

[0087] (Table 1) Table 1: Material balance of cocoa butter during the extraction and filtration processes. Material balance percentages are calculated based on a comparison with the percentage derived from cocoa nibs (100%). TIFF2026511937000001.tif64170

[0088] In addition to reducing the fat content in the cocoa extract, the distribution of the bioactive alkaloids theobromine and caffeine was monitored by HPLC. The results are shown in Tables 2 and 3.

[0089] (Table 2) Table 2: Material balance of alkaloids (caffeine and theobromine) during the extraction and filtration processes. Percentage balance calculations refer to cocoa nibs assumed to be 100%. TIFF2026511937000002.tif50170

[0090] (Table 3) Table 3: Alkaloid (caffeine and theobromine) content determined by HPLC in individual intermediate products from the extraction and filtration processes. TIFF2026511937000003.tif51170

[0091] Table 3 shows that during the extraction process, the alkaloids were concentrated in the hydrophilic liquid phase (1.11 g / 100 g caffeine and 5.92 g / 100 g theobromine relative to the defatted dry material). As explained by the mass balance, the alkaloids permeated well through the filtration membrane.

[0092] The balance of cocoa flavanols (DP1-DP7) during the extraction and filtration processes was analyzed by HPLC and fluorescence detection.

[0093] The results are summarized in Table 4, showing that flavanols with low degrees of polymerization (DP1-DP3) could be extracted well, especially with aqueous solvents, while flavanols with higher degrees of polymerization (i.e., DP4-DP7) were mostly found in the solid phase.

[0094] Similarly, during the filtration process, approximately 25-37% of the initial flavanol DP1-DP4 content in the cocoa nibs is recovered in the permeate, while the yield of flavanol DP5-DP7 decreases sharply to 8-15%.

[0095] (Table 4) Table 4: Mass balance of cocoa flavanols (DP1-DP7) during the extraction and filtration processes, as determined by HPLC / fluorescence detection. TIFF2026511937000004.tif25577

[0096] Table 5 lists the cocoa flavanol content in the fat-free dry material of each intermediate product. It is shown that the cocoa flavanol was mostly found in the hydrophilic liquid phase and was effectively recovered and concentrated in the permeate after the filtration step. In this experiment, the total yield of cocoa flavanol in the permeate (filtrate) was 27% relative to the total cocoa flavanol content in the cocoa nibs (assumed to be 100%).

[0097] (Table 5) Table 5: Total cocoa flavanol content (DP1-DP7) determined by HPLC / fluorescence detection. TIFF2026511937000005.tif51170

[0098] Table 6 shows the nutritional balance (carbohydrates, dietary fiber, and protein) in the individual intermediate products during the extraction and filtration processes. Percentage balance calculations refer to cacao nibs assumed to be 100%.

[0099] (Table 6) Table 6: Relative content of nutrients (carbohydrates, dietary fiber, protein) in individual intermediate products during the extraction and filtration process, as determined by 8 major nutrient analysis. TIFF2026511937000006.tif43170

[0100] During the extraction process, it was shown that the major components of carbohydrates, dietary fiber, and protein were not extracted by water and were largely retained in the solid phase. In the filtration step, carbohydrates and protein largely permeated into the filtrate, while dietary fiber was evenly distributed between the retaining liquid (i.e., P / DF concentrated powder) and the filtrate. A protein yield of 19% was observed in the permeate relative to the initial content in the cocoa nibs.

[0101] Adsorption / desorption of cocoa flavanols After the filtration process, the permeate (filtrate) was subjected to an adsorption / desorption process. For this purpose, a glass column was prepared and packed with a polymer adsorbent (AmberLite® XAD® 7HP, commercially available from DuPont®). The adsorbent used was a nonionic aliphatic acrylic resin, and its volume was measured by nitrogen BET method to approximately 520 m³. 2 The adsorbent had a surface area of ​​approximately 0.95 cc / g and a total pore volume of approximately 0.95 cc / g. The adsorbent was washed and conditioned with deionized water, and the permeate was pumped into the column, resulting in contact with the nonionic macroporous resin. The column was washed with deionized water before loading. Subsequently, the product was desorbed from the nonionic macroporous resin by elution in an aqueous alcohol solution using a polar gradient (60-80% ethanol). As a result, cocoa flavanol was released from the adsorbent, and this was pumped out of the column using a solvent to obtain a polyphenolic extract. Next, the obtained concentrated polyphenol fraction was concentrated in an evaporator and treated by spray drying to obtain a polyphenolic concentrate in powder form. Similarly, the non-adsorbed fraction was collected, concentrated in an evaporator, and treated by spray drying to obtain a powder, resulting in a theobromine concentrated powder.

[0102] Similar to the measurements described above for the extraction and filtration processes, individual fractions were analyzed for their alkaloid, flavanol, and nutrient content.

[0103] Table 7 shows the yields and distributions of the alkaloids caffeine and theobromine in the individual intermediate and final products across all three processing stages.

[0104] (Table 7) Table 7: Relative mass balance of alkaloids (caffeine and theobromine) during the extraction, filtration, and adsorption / desorption stages. Percentage balance calculations refer to cocoa nibs assumed to be 100%. TIFF2026511937000007.tif71170

[0105] These results indicate that while the majority of theobromine is found in the corresponding concentrated extract, caffeine is distributed almost equally between the theobromine concentrated extract and the polyphenolic concentrate. The concentration of theobromine in the theobromine concentrated extract was determined to be 4.5 g per 100 g of defatted dry material.

[0106] Table 8 shows the content and distribution of cocoa flavanols (DP1-DP7) throughout the adsorption / desorption process, compared to the permeate obtained in the filtration step. It was shown that 75% of the flavanol content in the filtrate could be recovered in the polyphenol concentrate, but at the same time, the ratio of higher-order oligomers (DP6 and DP7) decreased, and these were mostly found in the theobromine concentrate extract. A favorably high content of monomeric flavanol (DP1) of 10.5 g per 100 g of defatted dry material was achieved, which corresponds to a content of 23.4% by weight of the total content of flavanols with polymerization degrees 1-7 (DP1-DP7).

[0107] (Table 8) Table 8: Content distribution and yield of adsorbed / desorbed cocoa flavanols (DP1-DP7), determined by HPLC / fluorescence detection, compared to the content in the permeate (filtrate) assumed to be 100%. TIFF2026511937000008.tif249170

[0108] Table 9 summarizes the cocoa flavanol content in the defatted dried material of each intermediate and final product. These results indicate that the method of the present invention yields a polyphenolic extract with a favorably high concentration of cocoa flavanol, at 44.8 g per 100 g of defatted dried material.

[0109] (Table 9) Table 9: Total cocoa flavanol content (DP1-DP7) determined by HPLC / fluorescence detection. TIFF2026511937000009.tif78170

[0110] The total yield of cocoa flavanols in the polyphenolic concentrate was 20.3% relative to the cocoa flavanol content present in the starting material, i.e., cocoa nibs. Therefore, it has become clear that the method of the present invention simultaneously enables the provision of an extract with a high concentration of flavanols, a high ratio of monomeric and dimeric flavanols, and an excellent yield of total polyphenols relative to the total polyphenol content in the cocoa fruit components.

[0111] To further characterize the extract obtained by this method, the relative content of nutrients (carbohydrates, dietary fiber, protein) is determined by calculating the calorific value and carbohydrate content (according to Regulation (EU) No. 1169 / 2011), as well as the total fat content (by gravimetric method (Weibull-Stoldt) according to Norm (ASU) L44.00-4, 1985), saturated fatty acid content (by GC-FID method according to DGF C-VI 11e(18) / 10a(00) modified according to DGF C-VI 11e(98)), dietary fiber content (by enzyme-gravimetric method according to Norm (ASU) L00.00-18, 1997-01), protein content (by titration Kjeldahl method), and water / dry substance content (according to Norm (ASU) The analysis was performed according to the "Eight Major Analyses," including the determination of total ash content (by gravimetric method according to L44.00-4, 1985), sodium content (by Flame AAS), and sugar content (by HPLC-RI). The results for carbohydrates, dietary fiber, and protein are shown in Table 10, along with the content of intermediate products from the filtration process.

[0112] (Table 10) Table 10: Relative content of carbohydrates, dietary fiber, and protein in intermediate and final products. TIFF2026511937000010.tif64170

[0113] These results indicate that a significant proportion of the nutrients found in the permeate are separated via the non-adsorbed fraction, which yields theobromine concentrate. When considering the content relative to the defatted dry material, the polyphenolic concentrate is notably composed of protein, with 40.1g per 100g of defatted dry material, followed by carbohydrates at 4.7g per 100g, and dietary fiber at 1.5g per 100g. (Example 2)

[0114] In Example 2, 3387 kg of fermented and sterilized unroasted cocoa nibs (origin: Peru) were filtered at a temperature of 10±2°C using a membrane with a pore size of 0.1 μm and a filter area of ​​30 m².2 The extraction process was the same as in Example 1, except that the hydrophilic extract was subjected to a filtration step using a dynamic cross-flow filtration system with a ceramic membrane filter disc having [specific properties]. Furthermore, in contrast to Example 1, in Example 2, the P / DF concentrated extract and the non-adsorbed permeate were not subjected to powdering.

[0115] (Table 11) Table 11: Material balance of cocoa butter throughout the extraction process. Material balance percentages are calculated based on a comparison with the 100% percentage derived from cocoa nibs. TIFF2026511937000011.tif43170

[0116] The yields of the physiologically active alkaloids theobromine and caffeine were monitored by HPLC. The results are shown in Table 12.

[0117] (Table 12) Table 12: Mass balance of alkaloids (caffeine and theobromine) during the extraction, filtration, and adsorption / desorption stages. Percentage balance calculations refer to cocoa nibs assumed to be 100% (nm = not measured). TIFF2026511937000012.tif105170

[0118] Table 13 shows the mass balance of cocoa flavanols (DP1-DP7) as determined by HPLC throughout the extraction, filtration, and adsorption / desorption processes.

[0119] The adsorption / desorption process is shown to enable high yields of flavanols containing DP1-DP4 in polyphenolic powders, ranging from 48% to 60% relative to the filtrate. For flavanols containing DP5 or higher, the yield is even higher, at 59%-68%. Overall, a yield of 55% was achieved across all DP1-DP7 components.

[0120] Throughout all processing steps (extraction, filtration, and adsorption / desorption), the flavanol (DP1-DP7) yield was calculated to be 20%.

[0121] (Table 13) Table 13: Mass balance of cocoa flavanols (DP1-DP7) during the extraction and filtration processes, as determined by HPLC / fluorescence detection (nm = not measured). TIFF2026511937000013.tif252170

[0122] Table 14 summarizes the cocoa flavanol content in the defatted dried material of each intermediate and final product. These results show that the method of the present invention provides a polyphenolic powder containing a favorably high concentration of cocoa flavanol, 44.4 g per 100 g of defatted dried material. Advantageously, the polyphenolic powder showed a monomeric flavanol (DP1) content of 9.70 g per 100 g of defatted dried material.

[0123] (Table 14) Table 14: Total cocoa flavanol content (DP1-DP7) determined by HPLC / fluorescence detection. TIFF2026511937000014.tif71170 (Example 3)

[0124] In Example 3, 755 kg of (partially) defatted Lavado cocoa was subjected to the same extraction process and analysis as in Example 2. As shown in Table 15, in contrast to Examples 1 and 2, cocoa butter was not recovered via the hydrophobic liquid phase.

[0125] (Table 15) Table 15: Material balance of cocoa butter throughout the extraction and filtration process. Material balance percentages are calculated based on a comparison with 100% of the percentage derived from cocoa. TIFF2026511937000015.tif64170

[0126] The yields of the physiologically active alkaloids theobromine and caffeine were monitored by HPLC, and the results are shown in Table 16. Compared to Examples 1 and 2, higher yields of caffeine and theobromine were determined in both filtration and adsorption / desorption.

[0127] (Table 16) Table 16: Mass balance of alkaloids (caffeine and theobromine) during the extraction, filtration, and adsorption / desorption stages. Percentage balance calculations refer to starting materials assumed to be 100% (nm = not measured). TIFF2026511937000016.tif105170

[0128] Table 17 shows the mass balance of cocoa flavanols (DP1-DP7) throughout the extraction, filtration, and adsorption / desorption steps, as determined by HPLC.

[0129] In this experiment, 53% of the flavanols (DP1-DP7) were extracted into the hydrophilic liquid phase, while the remaining amount remained in the cocoa powder. Flavanols with low degrees of polymerization (DP1-DP3) were effectively extracted with the solvent, but flavanols DP4-DP7 were mostly found in the cocoa powder. During the filtration process, approximately 48%-72% of the initial content of flavanols DP1-DP4 was recovered in the permeate. For flavanols DP5-DP7, the yield in the permeate was calculated to be 11%-33%.

[0130] (Table 17) Table 17: Mass balance of cocoa flavanols (DP1-DP7) during the extraction and filtration processes, determined by HPLC / fluorescence detection (nm = not measured). TIFF2026511937000017.tif246170

[0131] Table 18 summarizes the cocoa flavanol content in the defatted dry material of the individual intermediate and final products of Example 3. As shown in the results, extraction resulted in a concentration of flavanols in the cocoa extract, from an initial 15.8 g per 100 g ffTS in defatted cocoa to 20.04 g per 100 g ffTS in the dry hydrophilic phase. Further concentration of flavanols was achieved in the permeate during filtration. Finally, a polyphenolic powder containing a very high concentration of cocoa flavanols, 88.2 g per 100 g defatted dry material, was obtained. These results indicate that the use of unfermented cocoa beans as the starting material and / or effective separation of cocoa butter (which may adversely affect adsorption efficiency) have a positive impact on both the flavanol yield throughout the processing steps and the concentration of flavanols in the final polyphenolic powder.

[0132] (Table 18) Table 18: Total cocoa flavanol content (DP1-DP7) determined by HPLC / fluorescence detection. TIFF2026511937000018.tif71170

[0133] Further examination of the flavanol (DP1-DP7) concentrations between individual diafiltration stages (each with an initial cocoa extract content of 15%) revealed that diafiltration substantially improved the yield of flavanols in the permeate.

[0134] In light of the above disclosures, many other features, modifications, and improvements will become apparent to those skilled in the art.

Claims

1. A method for obtaining an extract from the constituent materials of cocoa fruit, a) A step of subjecting the cacao fruit components to wet grinding in a polar solvent, separating the liquid phase from the wet grinding mixture to obtain a hydrophilic extract, b) A step of optionally separating residual solids from the hydrophilic extract, c) A step of adsorbing a hydrophilic extract onto a nonionic macroporous resin, d) A step of obtaining a polyphenolic extract by desorbing a hydrophilic extract from a nonionic macroporous resin using a water-miscible organic solvent, e) A step of concentrating the polyphenolic extract to obtain a polyphenolic concentrate. Methods that include...

2. Step a) is, a1) A step of adding a polar solvent to the cacao fruit components to form a suspension, a2) A step of wet grinding the suspension, a3) A step of subjecting the suspension to heat treatment at a temperature of 70°C or lower, a4) A step of separating the suspension into a hydrophilic liquid phase (heavy phase), a hydrophobic liquid phase (light phase), and a solid phase. Includes, The method according to claim 1, wherein the hydrophilic liquid phase comprises a hydrophilic extract as a major component and residual solids as a trace component, the hydrophobic liquid phase comprises cocoa butter as a major component and solids and / or a hydrophilic solvent as a trace component, and the solid phase comprises cocoa powder and a hydrophilic solvent.

3. The polar solvent is selected from water, C １ to C ８ alcohol, C ２ to C ８ ketone, C ３ to C ７ ester, C ２ to C ８ ether, C ４ to C １０ lactate, and mixtures thereof, optionally containing an acid, preferably acetic acid or citric acid, and / or Water-miscible organic solvents, C １ ~C ８ Alcohol, C ３ ~C ５ Ketones, C ３ ~C ５ Ester, C ２ ~C ４ Ether, C ２ ~C ５ Selected from aldehydes and mixtures thereof, preferably selected from methanol, ethanol, isopropanol, or acetone. The method according to claim 1 or claim 2.

4. The method according to any one of claims 1 to 3, wherein the cocoa fruit constituents in step a) include unfermented and unroasted cocoa beans, preferably unfermented and unroasted cocoa beans that have not been dried.

5. The method according to any one of claims 1 to 4, wherein in step a), the cocoa fruit constituent material is subjected to one or more wet grinding steps to an average particle size of 100 μm or less, preferably 50 μm or less, and more preferably 20 μm or less.

6. The method according to any one of claims 1 to 5, wherein the cacao fruit components and their extracts are not exposed to temperatures higher than 70°C.

7. The method according to any one of claims 1 to 6, wherein in step e), the polyphenolic concentrate is subjected to a concentration or evaporation step and an optional spray drying step to obtain a polyphenolic powder.

8. A nonionic macroporous resin, at least 400 m ２ The method according to any one of claims 1 to 7, having a surface area determined by the nitrogen BET method at a density of 0.90 cc / g, and / or a total pore volume determined by the nitrogen BET method at a density of 0.90 cc / g.

9. The method according to any one of claims 1 to 8, wherein in step b), residual solids are separated from the hydrophilic extract by filtration, preferably by cross-flow filtration and / or by filtration through a membrane filter having an absolute pore size rating of 0.45 μm or less, preferably 0.06 to 0.4 μm.

10. The method according to claim 9, wherein the residual solids are further processed by at least a concentration step to obtain a concentrated protein / dietary fiber extract.

11. The method according to any one of claims 1 to 10, wherein the non-adsorbed fraction of the hydrophilic extract in step c) is further treated by at least a concentration step to obtain a theobromine concentrated extract.

12. A polyphenolic extract obtained by the method described in any one of claims 1 to 11, preferably having the following characteristics: Total flavanol content determined by HPLC, at least 25 g per 100 g of defatted dry material, preferably at least 30 g per 100 g of defatted dry material, more preferably at least 50 g per 100 g of defatted dry material, and particularly preferably at least 53 g per 100 g of defatted dry material; The content of monomeric flavanol (DP1) determined by HPLC, at least 18% by weight, preferably at least 20% by weight, and more preferably at least 24% by weight, relative to the total content of flavanols having a degree of polymerization of 1 to 7 (DP1 to DP7); and Protein content of less than 42g per 100g of defatted dry material, carbohydrate content of less than 6g per 100g of defatted dry material, and dietary fiber content of less than 2g per 100g of defatted dry material; A polyphenolic extract that satisfies at least one of the following conditions.

13. A protein-concentrated extract obtained by the method described in claim 10.

14. Theobromine concentrated extract obtained by the method according to claim 11.

15. A food composition, supplement, medical composition, or cosmetic composition comprising the extract described in claims 12 to 14.

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