Cocoa extraction method and extract obtained by method

Abstract

The present invention describes a process for obtaining an extract from a cocoa fruit component, comprising the steps of: a) wet milling the cocoa fruit component in a polar solvent and separating a liquid phase from the wet-milled mixture to obtain a hydrophilic extract; b) optionally separating residual solids from the hydrophilic extract; c) allowing the hydrophilic extract to be adsorbed on a nonionic macroporous resin; d) desorbing the hydrophilic extract from the nonionic macroporous resin with a water-miscible organic solvent to obtain a polyphenol extract; and e) concentrating the polyphenol extract to obtain a polyphenol concentrate. In further aspects, an extract obtained by the aforementioned method is described as well as a food composition, a food supplement, a pharmaceutical composition or a cosmetic composition comprising the aforementioned extract.

Description

TECHNICAL FIELD

[0001] The present invention relates to an improved process for obtaining polyphenol, theobromine and protein rich extracts from cocoa fruit constituents using phase separation and adsorption / desorption techniques. In certain embodiments, the present invention relates to extracts obtained by the above process and compositions comprising the same. BACKGROUND

[0002] Cocoa (Theobroma cacao L.) is an important source of recognized healthy ingredients, including minerals, vitamins, and especially polyphenols (e.g. catechins, flavanol glycosides, anthocyanins and procyanidins). The main polyphenolic compounds present in cocoa beans are monomeric flavonoids such as (+)-catechin, (-)-epicatechin and oligomeric flavonoids such as procyanidins, which are known to reduce the risk of cardiovascular events, exerting hypotensive 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] As cocoa beans are one of the best known sources of dietary polyphenols, many extraction processes have been developed to obtain polyphenol-rich extracts from cocoa beans.

[0004] An exemplary process for extracting cocoa polyphenols from cocoa beans is disclosed in US 8 451 045 B2. US 6 159 451 A discloses a process for preparing a cocoa bean shell extract having improved inhibitory activity on glucosyltransferase in the prevention of dental caries.

[0005] In general, it is desirable to provide highly concentrated polyphenol extracts in the form of powders having the highest possible purity.

[0006] In this context, EP 2 071 961 B1 discloses a process for obtaining polyphenol-rich cocoa powder extracts, characterized in that they are obtained by subjecting fresh de-pulped cocoa seeds to blanching, drying and defatting by pressing, stabilizing the defatted product, grinding and subsequent solid-liquid extraction and liquid-liquid extraction and concentration steps.

[0007] WO 2005 / 115160 A1 and EP 1 748 700 A1 disclose a method for obtaining a cocoa polyconcentrate by subjecting unfermented cocoa beans to a blanching step, drying the unfermented cocoa beans, subjecting the dried unfermented cocoa beans to a particle size reduction treatment, extracting polyphenols from the dried unfermented cocoa beans intermediate to obtain a cocoa polyphenol extract and extracted solids, and concentrating the cocoa polyphenol extract to obtain a cocoa polyphenol concentrate, wherein the method further comprises a defatting step which is carried out by pressing the cocoa bean material prior to extraction.

[0008] However, there is still room for improvement in the overall polyphenol yield of these methods based on the initial polyphenol content in the raw material. For example, EP 2 071 961 B1 discloses an overall polyphenol yield of 11.93%.

[0009] Furthermore, it is not trivial to provide a polyphenol extract which is not only highly concentrated but also ensures a good bioavailability at the same time. Studies have shown that monomeric flavanol is more easily absorbed in the gut than polymeric proanthocyanidins and thus exhibits a higher antioxidant activity in vivo. However, the liquid-liquid extraction step can lead to an oligomer distribution favoring higher oligomers with a degree of polymerization (DP) of 3 or higher (see, for example, EP 1 913 821 A1). Moreover, a large number of processing steps, especially those involving thermal and / or mechanical energy input, can lead to degradation, epimerization and oxidative polymerization of polyphenols (see, for example, J. Alean et al., Journal of Food Engineering, 2016, 189, 99-105).

[0010] In order to avoid the use of mechanical pressing for separating the fat (i.e. cocoa butter) and to minimize the thermal load applied to the cocoa bean material, other cocoa bean processing methods have been developed to preserve higher contents of valuable heat-sensitive ingredients.

[0011] For example, WO 2010 / 073117 A1, EP 3 114 940 A1, EP 3 114 941 A1, EP 3 114 942 A1 and EP 3 114 939 A1 disclose a method of processing cocoa beans comprising forming a suspension comprising cocoa beans or cocoa nibs and water, wet milling the suspended cocoa beans or cocoa nibs, heating the suspension, and decanting the suspension such that the suspension separates into an aqueous phase, a fat phase and a solid phase, to avoid cocoa butter liquefaction and formation of chocolate liqueurs during mechanical processing, which method applies a lower thermal load to the cocoa mass compared to conventional methods and thus preserves higher contents of polyphenols, antioxidants and / or vitamins.

[0012] Advantageously, these processes envisage the manufacture of chocolate products by using a chocolate manufacturing kit comprising cocoa butter, cocoa powder, cocoa flavour and polyphenol powder extract, so that the polyphenol extract can be fully integrated into the chocolate production process. Here, the polyphenol powder is obtained by subjecting an aqueous phase to a de- aromatisation step, a concentration step (typically evaporation of excess water) and optionally spray-drying. However, it has been found that these processes are not suitable to provide a high polyphenol-enriched powder having a total flavanol content higher than 10 g / 100 g of dry defatted matter, as they usually result in a content of 2 to 8 g / 100 g of dry defatted matter depending on the variety.

[0013] In view of the above, it would still be desirable to provide a process that increases the yield of total polyphenols based on the total polyphenol content in the cocoa fruit ingredient and produces a polyphenol extract having both a high concentration of flavanols and a high proportion of monomeric and dimeric flavanols.

[0014] In addition to polyphenols, cocoa and cocoa products contain significant amounts of methylxanthines, i.e. theobromine (known to be a brain stimulant, having diuretic effects, potentially lowering blood pressure) and caffeine, which represent about 2.4-3.6% of the dry defatted cocoa composition. A commonly used process for extracting theobromine from cocoa beans typically involves extraction with a chlorinated alkane, such as chloroform or dichloromethane, as shown in US 1,925,326 A. However, the use of such extraction solvents is not recommended, not only from an environmental point of view, but also because the residual presence of such solvents in food products is undesirable and requires extensive removal steps.

[0015] Therefore, it would be desirable to efficiently extract these ingredients to provide additional extracts in the value chain of cocoa bean processing, for the preparation of food compositions, food supplements, pharmaceutical compositions or cosmetic compositions. SUMMARY

[0016] The present invention solves this object by the subject-matter of the claims defined herein. The advantages of the present invention will be further explained in the following sections, and further advantages will become apparent to the skilled person upon consideration of the disclosure of the present invention.

[0017] In one aspect, the present invention relates to a process for obtaining an extract from a cocoa fruit ingredient, comprising the steps of: a) wet-milling the cocoa fruit ingredient in a polar solvent and separating a liquid phase from the wet-milling mixture to obtain a hydrophilic extract; b) optionally separating residual solids from the hydrophilic extract; c) adsorbing the hydrophilic extract on a non-ionic macroporous resin; d) desorbing the hydrophilic extract from the non-ionic macroporous resin with a water-miscible organic solvent to obtain a polyphenol extract; and e) concentrating the polyphenol extract to obtain a polyphenol concentrate.

[0018] In another aspect, the present application relates to the polyphenol extract obtained by the above-mentioned method, preferably fulfilling at least one of the following characteristics: total flavanol content determined by HPLC of preferably at least 25 g / 100 g dry defatted matter, more preferably at least 30 g / 100 g dry defatted matter, further preferably at least 45 g / 100 g dry defatted matter, even more preferably 50 g / 100 g dry defatted matter, particularly preferably at least 53 g / 100 g dry defatted matter, most preferably at least 75 g / 100 g dry defatted matter; content of monomeric flavanols (DP1) determined by HPLC of at least 18 wt.%, preferably of at least 20 wt.%, more preferably of at least 24 wt.% based on the total content of flavanols with a degree of polymerization of 1 to 7 (DP1 to DP7); and protein content of less than 42 g / 100 g dry defatted matter, carbohydrate content of less than 6 g / 100 g dry defatted matter, dietary fiber content of less than 2 g / 100 g dry defatted matter, wherein the protein, carbohydrate and dietary fiber content can be determined by methods known in the art.

[0019] In another aspect, the present application relates to the protein / dietary fiber-enriched extract obtained by the above-mentioned method, wherein the residual solids are further processed by at least one concentration step, preferably spray-drying.

[0020] In another aspect, the present application relates to the protein-enriched extract obtained by the above-mentioned method, wherein the non-adsorbed portion of the hydrophilic extract in step c) is further processed by at least one concentration step.

[0021] In yet another aspect, the present application relates to a food composition, a food supplement, a pharmaceutical composition or a cosmetic composition comprising the aforementioned extract. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 An exemplary method for obtaining an extract from cocoa fruit components according to the present application is shown.

[0023] Figure 2 An exemplary method for making chocolate using the extract provided by the present application is shown. DETAILED DESCRIPTION

[0024] Extraction and fractionation methods

[0025] In a first embodiment, the present application relates to a method for obtaining an extract from a cocoa fruit component, comprising the steps of: a) wet-milling the cocoa fruit component in a polar solvent and separating the liquid phase from the wet-milling mixture to obtain a hydrophilic extract; b) optionally separating residual solids from the hydrophilic extract; c) adsorbing the hydrophilic extract on a non-ionic macroporous resin; d) desorbing the hydrophilic extract from the non-ionic macroporous resin with a water-miscible organic solvent to obtain a polyphenol extract; and e) concentrating the polyphenol extract to obtain a polyphenol concentrate.

[0026] Surprisingly, it was found that the order of the processing steps a) to e) increases the yield of total polyphenols based on the total polyphenol content in the cocoa fruit component and results in a polyphenol extract with both a high concentration of flavanols and a high proportion of monomeric and dimeric flavanols. Furthermore, since this method does not require the use of organic solvents which are not compatible with foodstuffs (such as hexane or halogenated alkanes such as chloroform, dichloromethane or carbon tetrachloride), it can be directly integrated into the preparation of cocoa products suitable for consumption (such as chocolate, cocoa beverages, cocoa-based desserts, food supplements, etc.) while minimizing the necessary processing steps.

[0027] As used herein, the expression "cocoa fruit component" refers to one or more components of a Theobroma cacao pod, wherein typically about 30 to 50 seeds of the fruit (i.e. cocoa beans) surrounded by pulp and mucilage are enclosed in the cocoa pod shell. Thus, such a cocoa fruit component can include cocoa beans, cocoa bean shell (CBS) material, cocoa pulp / mucilage and / or cocoa pod shell material. In a preferred embodiment of the present application, the cocoa fruit component subjected to step a) comprises cocoa beans, which can also include cocoa nibs.

[0028] In Figure 1 A non-limiting example of the method of the present application is illustrated in the flowchart shown below.

[0029] For the purposes of the present application, the cocoa beans or cocoa nibs can be fermented or unfermented, dried or undried, roasted or unroasted, and / or de-pulped or not de-pulped.

[0030] However, the total polyphenol content of fresh cacao beans is typically in the range of about 12-20% of the defatted dry matter, and about 5% for fermented cacao beans. In general, the polyphenol content in cacao beans depends on their origin and the processing (fermentation, drying treatment, alkalization, roasting, etc.). This can have a significant impact on the polyphenol content and thus on the quality of the final product. Since up to 70-80% of the polyphenols can be degraded during fermentation (see e.g. US 2004 / 0096566 Al), the cacao fruit ingredient subjected to step a) preferably comprises or consists of non-fermented cacao beans. The expression "fermented" generally refers to any activity or process involving the enzymatic or metabolic breakdown (digestion) of organic matter by microorganisms, including anaerobic and aerobic processes, as well as combined or sequential processes involving one or more anaerobic and / or aerobic stages. The term "non-fermented" as used herein refers to the degree of fermentation of the cacao beans. As known to the skilled person, the degree of fermentation is expressed by the fermentation index (see Shamsuddin and Dimick, Qualitative and Quantitative Measurements of Cacao Bean Fermentation in Proceedings of the Cacao Biotechnology Symposium, edited by P.S. Dimick, pp. 55-74, The Pennsylvania State University, 1986). In general, the fermentation index of non-fermented beans is less than 0.5. It is further preferred that step a) is performed on the cacao beans immediately after opening the cacao fruit pods and separating the cacao beans from the pod shells and optionally removing the pulp / mucilage. Another example of preferably cacao beans includes lavado beans, which are non-fermented beans (typically of reddish color), prepared by removing the cacao beans from the mature pods, washing with water to remove the pulp, and drying (including but not limited to sun drying).

[0031] Furthermore, polyphenols are generally considered as heat sensitive ingredients. Therefore, in order to preserve the original polyphenol content of the cacao beans as much as possible, the cacao fruit ingredient subjected to step a) preferably comprises cacao beans which have not been subjected to a drying and / or roasting process.

[0032] In another preferred embodiment, the non-fermented cacao beans can be pre-treated in an incubation step with or without cacao pulp and mucilage prior to step (a). This incubation step is characterized by inhibiting the sprouting of the cacao beans by certain physical and / or chemical pre-treatments shortly after removal of the cacao beans (with or without pulp and mucilage) from the pods.

[0033] In particular, germination of the cocoa beans can be inhibited by incubating freshly harvested cocoa beans in an incubation medium at an elevated temperature, for example at a temperature between 10 °C and 70 °C, preferably between 10 °C and 55 °C, for a period of 2 hours to 10 days, preferably 3 to 168 hours. As known to the skilled person, it is particularly preferred to transfer the cocoa beans (optionally together with the pulp / mucilage) into the incubation medium immediately after opening the pods, preferably under sterile conditions, to inhibit spontaneous fermentation as effectively as possible. The incubation medium is not particularly limited and can be an aqueous acidic solution, for example as used in the cocoa bean processing method disclosed in US 8,501256 B2, or an aqueous ethanol solution.

[0034] Furthermore, polyphenols are generally known to be heat sensitive ingredients. Therefore, in order to preserve the original polyphenol content of the cocoa beans as much as possible, the cocoa fruit ingredient subjected to step a) preferably comprises cocoa beans which have not been subjected to a drying and / or roasting process comprising a heat treatment above 100 °C or preferably above 70 °C.

[0035] In step a), the cocoa fruit ingredient is wet milled in a polar solvent and the liquid phase is separated from the wet milling mixture to obtain a hydrophilic extract.

[0036] The polar solvent used herein generally has a dielectric constant of 15 or more, preferably a dielectric constant of 50 or more, more preferably a dielectric constant of 80 or more. While the polar solvent can include polar protic solvents and polar aprotic solvents, it is preferred to use polar protic solvents. In preferred embodiments, the polar solvent is selected from 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 tert-butyl ether), C4-C 10 lactates and mixtures thereof. It is particularly preferred that the polar solvent is water or an aqueous ethanol mixture. It is further preferred that the polar solvent is free of halogenated hydrocarbons.

[0037] In preferred embodiments, the polar solvent comprises or consists of water. In other preferred embodiments, the polar solvent can comprise water in an amount of less than 95 wt.-% (e.g. less than 90 wt.-%, less than 80 wt.-%, less than 50 wt.-%, less than 30 wt.-%, less than 10 wt.-% or less than 5 wt.-%) in admixture with one or more organic solvents which are miscible or immiscible with water under the conditions of the extraction. It is to be understood that alternative aqueous liquids can also be used as the source of water, preferably a liquid selected from one or more of coffee, tea and a liquid having a water content of from 60 wt.-% to about 95 wt.-%, for example a fruit juice, a concentrated fruit juice material or milk.

[0038] In preferred embodiments, the polar solvent can comprise an acidic component (including but not limited to acetic acid, citric acid or other acids) which is added in addition to the acetic acid and / or citric acid formed by microbial activity in the beans, which can facilitate efficient extraction at low temperature and / or reduce oxidative degradation of polyphenols.

[0039] According to the present application, the cocoa bean ingredient is wet milled in the presence of a polar solvent to avoid or reduce the disruption of the cellular structure of the cocoa material due to high mechanical load or shear stress involved in dry milling methods conventionally employed in the art.

[0040] While not particularly limited, the weight ratio of polar solvent to cocoa bean ingredient in the suspension in which wet milling is performed is preferably between 1 : 1 and 6: 1, more preferably between 2: 1 and 4: 1, and particularly preferably about 3: 1, which can advantageously influence processability in subsequent steps (e.g. facilitating pumping, milling and easier phase separation).

[0041] The cocoa fruit ingredient is preferably wet milled to an average particle size of 100 pm or less, preferably 50 pm or less, more preferably 20 pm or less, preferably by a single or multiple wet milling steps. Reducing the cocoa fruit ingredient to such a size range greatly increases the exposed surface area of the particulate material, enabling it to be wetted more efficiently to improve 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.

[0042] The cocoa fruit ingredient (e.g. cocoa beans or cocoa nibs) is preferably soaked to enable the polar solvent to wet the solids better due to the increased available surface area of the solids as a result of the soaking treatment, preferably in at least one milling step. The method and equipment used for wet milling is not particularly limited, provided that undesirable emulsification caused by significant frictional heat generation or high mechanical forces is avoided. For example, when multiple milling steps are used, a coarse wet milling step (e.g. optionally using a different polar solvent) can be performed using a perforated disc mill, and the coarsely milled suspension can be pumped to a toothed colloid mill for a fine milling step. It will be appreciated that if multiple milling steps are employed, the polar solvent can be modified or changed between steps (e.g. by adding other solvents or acids). In preferred embodiments, soaking is achieved by cold milling (one or more steps) at a temperature of 2 °C to 25 °C.

[0043] In a preferred embodiment of the present application, step a) comprises the steps of al) adding a polar solvent to the cocoa fruit ingredient to form a suspension; a2) wet milling the suspension; a3) subjecting the suspension to heat treatment at a temperature of 70 °C or less; and a4) separating the suspension into a hydrophilic liquid phase (heavy phase) comprising as a main component a hydrophilic extract and as a minor component residual solids, a hydrophobic liquid phase (light phase) comprising as a main component cocoa butter and as a minor component solids and / or hydrophilic solvent, and a solid phase comprising cocoa powder and hydrophilic solvent. Steps al) and a2) can be performed on the basis of preparing and milling the suspension according to the above description. In step a3), the wet-milled suspension is subjected to heat treatment at a temperature of not more than about 70 °C to reduce the overall heat load and to prevent emulsification of the cocoa bean material. The heating temperature is preferably between 43 °C and 65 °C from the perspective of a good balance between cocoa butter yield and preservation of the desired flavours, such as aroma, antioxidants and / or vitamins. A heating temperature range of 45 °C to 50 °C is particularly preferred in terms of cocoa butter liquefaction and / or improved mechanical phase separation. Without being limited thereto, the heating of the wet-milled suspension can be performed by means of a scrap or tube heat exchanger.

[0044] Thereafter, in step a4), the suspension is separated into a hydrophilic liquid phase (heavy phase) comprising as a main component a hydrophilic extract and as a minor component residual solids, a hydrophobic liquid phase (light phase) comprising as a main component cocoa butter and as a minor component solids and / or hydrophilic solvent, and a solid phase comprising cocoa powder and hydrophilic solvent. Furthermore, the solid phase can comprise residual cocoa butter in an amount of at most 30 wt.%, preferably below 27 wt.%, more preferably below 20 wt.%, relative to the total dry weight.

[0045] Preferably, the mechanical particle separation can be achieved with means of centrifugal force, such as a decanter or a nozzle separator. For example, the suspension can be decanted to separate coarse or large or high mass solids from the liquid, which can then be further separated from smaller and / or fine solid particles, and / or the oil product can be separated from the non-oil product.

[0046] Improved separation between the hydrophilic liquid phase (heavy phase), the hydrophobic liquid phase (light phase) and the solid phase can be achieved with a multi-phase separation and recombination step. For example, the hydrophobic liquid phase obtained by the initial decanting step can be further filtered or centrifuged to separate remaining fine particles or water from the hydrophobic liquid phase, and the fine particles and polar solvent thus obtained can be recombined with the polar solvent and solid phase from the initial decanting step or in a subsequent processing stage of said phases.

[0047] After the three-phase separation, the phases can be processed separately to isolate, for example, cocoa butter (from the hydrophobic liquid phase), cocoa powder (from the solid phase) and cocoa aroma (from any of the three phases). Although not limited thereto, examples of independent processing of the phases and their combination are disclosed in WO 2010 / 073117 A1, EP 3 114 940 A1, EP 3 114 941 A1, EP 3 114 942 A1, EP 3 114 939 A1, EP 3 747 277 A1, EP 3 747 275 A1 and EP 3 747 276 A1.

[0048] According to the present application, the hydrophilic liquid phase obtained in step a) is further processed to extract a polyphenol concentrate.

[0049] At this stage, the hydrophilic liquid phase can optionally be removed or neutralized from the hydrophilic liquid phase if an acid was added in step a). Suitable removal methods are not particularly limited and can be carried out by any suitable method known in the art, for example by extractive rectification, reactive distillation, extraction (e.g. liquid-liquid extraction), emulsion-type liquid membrane process, salting-out or a combination thereof.

[0050] Generally, the hydrophilic liquid phase obtained in step a) (or step a4), respectively, comprises residual amounts of solids and cocoa butter. According to the present application, it is preferred to separate the residual solids from the hydrophilic extract to remove fine particles and cocoa butter residues that can adversely affect the adsorption process in step c). To this end, it is preferred to subject the hydrophilic liquid phase to a filtration step in step b). In a further preferred embodiment, the filtration process and clarification can be enhanced by the prior addition of a filter aid and / or a protein precipitant. 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 to the isoelectric point by the addition of an acid and / or a base to the isoelectric point to achieve isoelectric precipitation and improve the efficiency of the subsequent filtration. As examples of protein precipitants, carrageenan, agar, alginic acid, bentonite, pectin, colloidal silica, tannin or a combination thereof can be mentioned.

[0051] In a particularly preferred embodiment, filtration is performed using a cross-flow filtration with an absolute pore size rating of 0.45 pm or less, for example from 0.02 pm to 0.45 pm and / or by membrane filtration. Particularly preferred is the use of dynamic cross-flow filtration using ceramic membranes, wherein the cross-flow effect (= tangential overflow of the membrane surface) is generated by rotating filter discs rather than by pumping large amounts of liquid. Thus, efficient filtration can be performed continuously at a relatively high solids load without clogging the filter. It is further preferred to use membrane filters with an absolute pore size rating of 0.30 pm or less, for example membrane filters with a pore size rating of 0.25 pm or less, 0.22 pm or less, 0.20 pm or less or 0.10 pm or less. In some embodiments, it can be particularly preferred to use membrane filters with an absolute pore size rating of 0.20 pm or less. For example, it has been found that filtration using membrane filters with a pore size rating between 0.04 pm and 0.16 pm, more preferably between 0.06 pm and 0.10 pm, advantageously leads to a higher concentration of flavonols in the desorbed polyphenol extract without a significant reduction in the yield of flavonols.

[0052] In preferred embodiments, filtration can be performed at a temperature of from 3 °C to 50 °C, for example between 25 °C and 35 °C. However, it is particularly preferred to perform filtration at a temperature of from 4 °C to 15 °C or from 5 °C to 10 °C.

[0053] In particularly preferred embodiments, the diafiltration step is further integrated by introducing purified solvent (e.g. nanofiltered (NF) or reverse osmosis (RO) water) into the process fluid to facilitate extraction of the flavonoid compounds. Preferably, the volume of purified solvent is adjusted to 45% or less of the total volume of the feed product (cocoa extract, respectively).

[0054] The retentate (i.e. the residual solids) obtained after filtration can be further processed by at least one concentration step to obtain a protein-enriched extract, which can be achieved by evaporation of the residual polar solvent in one or more steps. The protein-enriched extract typically comprises a significant amount of dietary fiber and can thus also be understood as a protein- / dietary fiber-enriched extract (P / DF-enriched extract). Optionally, the protein-enriched extract thus obtained can be subjected to spray-drying to provide a cocoa-based, protein-enriched powder or a protein- / dietary fiber-enriched powder (P / DF-enriched powder), which represents an additional valuable and previously often discarded ingredient.

[0055] The permeate (i.e. the clarified hydrophilic extract) obtained in the filtration step typically contains less than 1 g / 100 g dry matter of cocoa butter. The major part of the flavanols and theobromine contained in the hydrophilic phase is typically still present in the permeate as well, while also a significant amount of amino acids is contained.

[0056] In step c), the resulting hydrophilic extract can optionally be gently heated (below 70°C, preferably below 60°C) and adsorbed onto a nonionic macroporous resin. For this purpose, a column containing a nonionic macroporous resin is prepared, washed, and appropriately conditioned using methods known to those skilled in the art, and the hydrophilic extract is pumped through the column, thereby adsorbing cocoa polyphenols onto the resin.

[0057] In a preferred embodiment, the nonionic macroporous resin comprises or is composed of cross-linked aliphatic polymers, aliphatic acrylic polymers, or polyacrylic polymers.

[0058] In terms of improving bonding ability and process efficiency, nonionic macroporous resins preferably have a molecular weight of at least 400 μm as determined by the nitrogen BET method. 2 The surface area per g, more preferably at least 450 m² 2 The surface area per g is more preferably at least 500 m². 2 / g surface area. Alternatively or in combination, the nonionic macroporous resin preferably has a total pore volume of at least 0.55 cc / g as determined by the nitrogen BET method, more preferably at least 0.80 cc / g total pore volume, even more preferably at least 0.85 cc / g total pore volume, and particularly preferably at least 0.99 cc / g total pore volume.

[0059] After adsorption, the resin can optionally be washed (e.g., with deionized water) to remove unwanted components. The washing step is preferably carried out at a temperature of 10 to 50°C. It is worth noting that the unadsorbed portion of the hydrophilic extract in step c) can be further processed by at least one concentration step to obtain a theobromine-rich extract.

[0060] In step d), the hydrophilic extract is desorbed from the nonionic macroporous resin by eluting the adsorbate with a water-miscible organic solvent to obtain a polyphenol extract.

[0061] 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. Optionally, the water-miscible organic solvent can be diluted with water, provided that the water content does not exceed 50% by volume. In a particularly preferred embodiment, an aqueous solution of ethanol (e.g., 60%-80% by volume ethanol) can be used. Further preferred is that the water-miscible organic solvent is free of halogenated hydrocarbons. In a preferred embodiment, an acidic component (including but not limited to inorganic acids, acetic acid, citric acid, or other acids) can be added to the water-miscible organic solvent.

[0062] It is worth noting that the loaded resin can be eluted with the same solvent but at different temperatures or with different solvents in succession. For example, it is conceivable that the alkaloids are first selectively eluted from the non-ionic macroporous resin using a first solvent having a high solubility for caffeine and / or theobromine, and then the polyphenolic ingredients are eluted using a second solvent, i.e. a water-miscible organic solvent. Depending on the desired quality of the individual extracts, the desorption process can be optimized accordingly.

[0063] Alternatively, the water-miscible organic solvent can be heated prior to elution in order to facilitate the removal of unwanted volatile compounds.

[0064] In step e), the polyphenol extract obtained in step d) is subjected to a concentration step to obtain a polyphenol 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. From the perspective of stabilizing the polyphenolic ingredients, such as flavanols, vacuum distillation / evaporation and spray drying in this order to obtain a polyphenol powder are preferred.

[0065] In order to preserve as many of the polyphenols contained in the original cocoa fruit ingredients as possible, the cocoa fruit ingredients and their extracts are preferably not subjected to temperatures above 70°C at least during steps a) to d), more preferably during steps a) to e), and further preferably throughout the entire extraction process.

[0066] Advantageously, the above-described method can be fully integrated into the process for the preparation of cocoa-based products for human consumption, such as chocolate, cocoa powder, cocoa butter, etc., without relying on unwanted solvents and provides for an excellent yield of cocoa polyphenols. In particular, a total polyphenol yield of 15% or more, such as a total polyphenol yield in the range of 15 to 30%, typically at least 20% of the polyphenol content in the cocoa beans (100%) can be achieved.

[0067] Cocoa extract

[0068] In a second embodiment, the present application relates to a polyphenol extract obtained by the method of the first embodiment described above.

[0069] Generally, the polyphenolic compounds originally present in the cocoa bean include mainly catechins ((-)-epicatechin, (+)-catechin, (+)-gallocatechin and (-)epigallocatechin), proanthocyanidins (procyanidins Bl to B5, procyanidin Cl, procyanidin D and higher oligomers and polymers, mainly homologues of epicatechin with 2 to 18 monomeric units), anthocyanins (i.e. cyanidin-3a-L-arabinoside and cyanidin-3-P-D-galactoside), flavanol glycosides (i.e. quercetin-3-0-a-D-arabinoside and quercetin 3-0-P-D-glucoside), eugenol amides and dideoxyeugenol amides.

[0070] The term "cocoa flavanols" or "flavanols" as used herein refers to the monomeric flavanols (-)-epicatechin and (+)-catechin with a degree of polymerization (DP) of DP1 to DP7 and their related oligomeric flavanols (proanthocyanidins). The determination of the individual contents based on the degree of polymerization (DP1 to DP7) and the total flavanol content can be determined by HPLC methods combined with fluorescence detection of commercially available standards (e.g. according to U. Bussy et al., Food Funct. 2020, 11 (1), 131-138).

[0071] According to a preferred embodiment, the polyphenol extract of the present application has a total flavanol content of at least 25 g / 100 g dry defatted matter, more preferably of at least 30 g / 100 g dry defatted matter, further preferably of at least 45 g / 100 g dry defatted matter, even further preferably of at least 50 g / 100 g dry defatted matter, particularly preferably of at least 53 g / 100 g dry defatted matter, most preferably of at least 75 g / 100 g dry defatted matter, as determined by HPLC. It is worth noting that the total flavanol content can exceed the currently available flavanol standards (e.g. NIST reference material 8403). The upper limit of the total flavanol content is not particularly limited and can be 100 g / 100 g dry defatted matter.

[0072] Additionally or alternatively, the polyphenol extract of the present application preferably shows a content of monomeric flavanols (DP1) of at least 18 wt.-%, more preferably at least 20 wt.-%, further preferably at least 24 wt.-%, as determined by HPLC, based on the total content of flavanols with a degree of polymerization of 1 to 7 (DP1 to DP7). It can be further preferred that the polyphenol extract of the present application preferably shows a content of dimeric flavanols (DP2) of at least 18 wt.-%, preferably at least 20 wt.-%, as determined by HPLC, based on the total content of flavanols with a degree of polymerization of 1 to 7 (DP1 to DP7). Thus, the polyphenol extract contains high amounts of monomeric and dimeric flavanols which show a particularly advantageous bioavailability. Additionally or alternatively, the polyphenol extract of the present application preferably exhibits a protein content of less than 42 g / 100 g dry matter free of fat (e.g. 5 to 41 g / 100 g dry matter free of fat), a carbohydrate content of less than 6 g / 100 g dry matter free of fat (e.g. 2 to 5 g / 100 g dry matter free of fat), and a dietary fiber content of less than 2 g / 100 g dry matter free of fat (e.g. 0.1 to 1.8 g / 100 g dry matter free of fat). The above-mentioned nutritional ingredients are characteristic for the polyphenol extract prepared by the process of the present application. The respective content of protein, carbohydrate and dietary fiber can be determined by nutritional analysis methods commonly used in the art, including the "Big 8" or "Big 12" nutritional analysis methods. The protein content can be determined based on the total nitrogen content by titration Kjeldahl method, e.g. using a conversion factor of 6.25 (see e.g. AOAC method 970.22). The term "dietary fiber" as used herein is understood to define indigestible carbohydrates and lignin, and the dietary fiber content can be determined according to methods well known to the person skilled in the art, e.g. by determining the difference in heat value according to the method prescribed by Regulation (EU) No 1169 / 2011. The content of (digestible) carbohydrates can be calculated from 100% minus the total fraction of moisture, ash, protein, fat, alcohol and insoluble organic fibers in the sample in %, wherein the fat content can be determined by e.g. the Weibull-Stoldt method.

[0073] In a third embodiment, the present application relates to a protein-enriched extract obtained by the process according to the first embodiment. As described above, the protein-enriched extract is derived from the retentate (i.e. the residual solids) obtained after filtration, which retentate is subjected to at least one concentration step to remove the polar solvent, and further comprises cocoa-derived dietary fibers. Optionally, the protein-enriched extract thus obtained can be subjected to spray-drying to provide a cocoa-based protein-enriched powder.

[0074] Typically, the protein content of the protein-enriched powder is at least 30 wt%, often more than 35 wt%, based on the total dry weight of the protein, the carbohydrate and the dietary fiber. The respective content can be suitably determined by methods known in the art, for example as the Kjeldahl method or the Dumas method, using a conversion factor of for example 6.25.

[0075] In a fourth embodiment, the present application relates to a theobromine-enriched extract obtained by the method according to the first embodiment. As mentioned above, the theobromine-enriched extract is derived from the non-adsorbed fraction of the hydrophilic extract in step c), which is further processed, preferably dried, by at least one concentration step to provide the extract in the form of a powder. The theobromine-enriched extract typically exhibits a theobromine content of at least 3 g / 100 g of defatted dry matter, more preferably a theobromine content of at least 4 g / 100 g of defatted dry matter, and particularly preferably a theobromine content of at least 5 g / 100 g of defatted dry matter.

[0076] Composition and use of the extract

[0077] In a fifth embodiment, the present application relates to a food composition, a food supplement, a pharmaceutical composition or a cosmetic composition comprising the extract according to the second to fourth embodiments, respectively, or comprising the polyphenol extract, the protein-enriched extract and / or the theobromine-enriched extract produced according to the first embodiment.

[0078] It will be appreciated that the various extracts obtained by the method of the present application can be used and / or sold separately or integrated into the cocoa bean processing method aimed at providing food and beverage compositions.

[0079] Exemplary food compositions according to the present application include cocoa-based products suitable for consumption in liquid and solid form (e.g. chocolate, cocoa-based desserts, cocoa beverages).

[0080] Furthermore, the extracts, preferably in the form of a powder, can be used in a building kit for cocoa-based products (e.g. chocolate, cocoa-based desserts, cocoa beverages), which kit can further comprise cocoa powder (including but not limited to cocoa powder provided by drying the solid phase obtained in step a), or a4), respectively), cocoa butter (including but not limited to cocoa butter provided by processing the hydrophobic liquid phase obtained in step a4), and / or cocoa aroma (including but not limited to cocoa aroma obtained by de-aromatization of the hydrophilic liquid phase obtained in step a4).

[0081] For example, it is conceivable to use polyphenol extracts, protein-rich extracts and / or theobromine-rich extracts, each preferably in powder form, in the methods of WO 2010 / 073117 A1, EP 3 114 940 A1, EP 3 114 941 A1, EP 3 114 942 A1, EP 3 114 939 A1, EP 3 747 277 A1, EP 3 747 275 A1 and EP 3 747 276 A1, in order to be able to further fine-tune the sensory and / or nutritional properties of the desired product.

[0082] Figure 2 An example of a method for preparing dark and milk chocolate based on a chocolate building kit comprising cocoa aroma, cocoa powder, cocoa butter and polyphenol extract, protein-rich extract and theobromine-rich extract produced according to the present application is shown. While not being limited thereto, the cocoa aroma can be obtained from de-aromatization of the hydrophilic phase and / or can be a roasted cocoa aroma obtained from drying / roasting of the cocoa powder. The components of the kit are mixed prior to performing the conching step. In comparison to the prior art methods, the polyphenol extract according to the present application can be used to increase the yield of polyphenols and to increase the bioavailability of the final product. The protein-rich extract (or protein / dietary fiber-rich extract, respectively) is a part of the above-mentioned prior art cocoa bean processing methods that has not been used before and can be added or re-introduced to alter the protein content in the final product and to ideally efficiently utilize the cocoa bean ingredients. Additional customization of the flavor or development of a flavor can be performed by adding one or more of sugar, sweetener, cocoa nibs and / or juice. Furthermore, the theobromine-rich extract can be selectively added or re-introduced to utilize the pharmacological effects of the alkaloid and / or to balance the bitterness of the cocoa-based product.

[0083] For the preparation of milk chocolate, it is preferred to further add milk powder prior to the conching step. Optionally, an emulsifier (e.g. lecithin) can be added prior to conching to reduce viscosity, control sugar crystallization and flow properties of the chocolate, and aid in the uniform mixing of the ingredients. In addition, additional ingredients and flavorings, such as vanilla, rum, etc., e.g. sea salt, iris, nuts and raisins, can be added prior to or during the conching step, as well as after conching. The conching process redistributes the flavor-producing substances in the dry cocoa into the fat phase, while removing unwanted acetic, propionic and butyric acids from the chocolate, reducing moisture, and resulting in a mellow flavor of the product. The temperature of the conche is controlled and varies depending on the different types of chocolate (from about 49°C for milk chocolate to as high as 82°C for dark chocolate). While dependent to some extent on temperature, the conching duration in the traditional chocolate manufacturing process is typically in the range of 16 to 72 hours for good results. The conching duration is preferably less than 16 hours, more preferably less than 12 hours, and typically 10 hours or less. Thus, the loss of desirable flavor characteristics and degradation of heat-sensitive ingredients, including the polyphenol fraction, is minimized.

[0084] For pharmaceutical compositions or dietary supplements, the extract of the present application can be administered in a form and at a dosage appropriate for the skilled person in the art, e.g. in capsules or tablets.

[0085] Due to the antioxidant properties of the polyphenolic ingredients, the polyphenol concentrate obtained in the present application can also be used in cosmetic applications, e.g. in shampoos, lotions, creams, balms, ointments, etc.

[0086] It will be appreciated that features of the first to fifth embodiments can be freely combined in any combination, except that at least some features are mutually exclusive in combination. DETAILED DESCRIPTION

[0088] Example 1

[0089] Extraction and phase separation

[0090] A quantity of 2569 kg of fermented, sterilized and unroasted cocoa nibs (origin: Peru) was mixed with water as the polar solvent in a ratio of about 1 :3 to form a suspension. Subsequently, the suspension was wet milled in multiple stages using a perforated colloid mill to an average particle size of less than 100 μιη and heated to a temperature of about 45°C to 60°C using a tube heat exchanger. Using a three-phase decanter, the two immiscible liquid phases, i.e. the hydrophilic and the hydrophobic liquid phase, were separated from each other, while the solid phase was discharged. After removal of the lighter oil phase and the solid phase, the hydrophilic liquid phase was used to provide a polyphenol extract, a theobromine-rich extract and a protein-rich extract according to the procedures described in the following sections.

[0091] Filtration

[0092] Initially, a filtration step was performed on the hydrophilic extract using a dynamic cross-flow filtration system with ceramic membrane filter discs (commercially available from Novoflow GmbH; filtration area: 3 m 2 ; membrane pore size: 0.2 pm) at a temperature of 5-10 °C. Diafiltration was performed by introducing 10% of the total volume of the hydrophilic extract feed with reverse osmosis water.

[0093] From the filtration of the aqueous cocoa extract, two products were obtained, one being the retentate mainly comprising proteins and dietary fibers, and the other being the filtrate mainly comprising the aqueous solvent, the cocoa flavanols, caffeine and theobromine, and proteins. The retentate was then concentrated in an evaporator and converted into a protein-rich powder or a protein- / dietary fiber-rich powder (hereinafter referred to as P / DF-rich powder), respectively, by spray drying.

[0094] Furthermore, as shown in Table 1, the filtration step effectively removed residual fat (i.e. cocoa butter) from the hydrophilic extract to avoid interference with the subsequent adsorption.

[0095] Table 1: Mass balance of cocoa butter during extraction and filtration. The calculation of the percentage of the mass balance is based on a percentage of 100% of the cocoa nibs.

[0096]

[0097] In addition to the reduction of the fat content in the cocoa extract, the distribution of the biologically active alkaloids theobromine and caffeine was monitored by HPLC, the results of which are shown in Tables 2 and 3.

[0098] Table 2: Mass balance of alkaloids (caffeine and theobromine) during extraction and filtration. The calculation of the percentage of the balance is based on a percentage of 100% of the cocoa nibs.

[0099]

[0100] Table 3: Content of alkaloids (caffeine and theobromine) in the intermediates during extraction and filtration as determined by HPLC.

[0101]

[0102] Table 3 shows that during the extraction process, the alkaloids were enriched in the hydrophilic liquid phase (1.11 g / 100 g caffeine and 5.92 g / 100 g theobromine based on the defatted dry matter). As shown by the mass balance, the alkaloids permeated well through the filtration membrane.

[0103] The mass balance of the cocoa flavanols (DP1-DP7) during extraction and filtration was analyzed by HPLC and fluorescence detection.

[0104] The results, summarized in Table 4, show that the lower degree of polymerization flavanols (DP1-DP3) are particularly well extracted with the aqueous solvent, while the higher degree of polymerization flavanols (i.e. DP4-DP7) are mainly present in the solid phase.

[0105] Similarly, during the filtration step, the initial content of DP1-DP4 flavanols in the cocoa nibs was recovered in the permeate, with a yield of about 25-37% in the permeate, while the yield of DP5-DP7 flavanols dropped dramatically to 8-15%.

[0106] Table 4: Mass balance of the cocoa flavanols (DP1-DP7) during extraction and filtration as determined by HPLC / fluorescence detection.

[0107]

[0108] Table 5 lists the cocoa flavanol content in the fat-free dry matter of the various intermediate products. The results show that the cocoa flavanols are mainly present in the hydrophilic liquid phase and are effectively recovered and enriched in the permeate after the filtration step. In this experiment, the total yield of cocoa flavanols in the permeate (filtrate) was 27% based on the total content of cocoa flavanols in the cocoa nibs (assumed to be 100%).

[0109] Table 5: Total content of cocoa flavanols (DP1-DP7) as determined by HPLC / fluorescence detection.

[0110]

[0111] Table 6 shows the balance of the nutritional values (carbohydrates, dietary fiber, protein) of the various intermediate products during extraction and filtration. The balance percentages were calculated based on the cocoa nibs, which were assumed to be 100%.

[0112] Table 6: Relative content of the nutritional components (carbohydrates, dietary fiber, protein) of the various intermediate products during extraction and filtration as determined by the analysis of the 8 core nutritional components.

[0113]

[0114] The study shows that during the extraction step, the major part of the carbohydrates, dietary fiber and protein are not extracted with water, but are mainly retained in the solid phase. During the filtration step, the carbohydrates and protein mainly permeate into the filtrate, while the dietary fiber is evenly distributed between the retentate (i.e. the P / DF-rich powder) and the filtrate. In the permeate, a protein yield of 19% was found based on the initial content in the cocoa nibs.

[0115] Adsorption / desorption of cocoa flavanols

[0116] Following the filtration step, the permeate (filtrate) is subjected to an adsorption / desorption step. To this end, a glass column is prepared and packed with a polymeric adsorbent (Amberlite™ XAD™ 7 HP, commercially available from DuPont™). The adsorbent used is a non-ionic aliphatic acrylic resin having a surface area of about 520 m 2 / g, and a total pore volume of about 0.95 cc / g, both as determined by nitrogen BET. After washing and conditioning the adsorbent with deionized water, the permeate is pumped into the column, thereby contacting the non-ionic macroporous resin. After loading, the column is washed with deionized water. Thereafter, the product is desorbed from the non-ionic macroporous resin using aqueous alcoholic solutions with a polarity gradient (60% to 80% ethanol). As a result, the cocoa flavanols are released from the adsorbent and can be pumped from the column together with the solvent to provide a polyphenol extract. The polyphenol-rich fraction obtained is then concentrated in an evaporator and processed by spray-drying to obtain a polyphenol concentrate in the form of a powder. Similarly, the non-adsorbed fraction is collected, concentrated in an evaporator and processed by spray-drying to a powder to obtain a theobromine-rich powder.

[0117] The content of alkaloids, flavanols and nutrients is analyzed for each fraction, similarly to the measurements described above for the extraction and filtration steps.

[0118] Table 7 shows the yield and distribution of alkaloids, caffeine and theobromine in the individual intermediates and final products throughout the three processing steps.

[0119] Table 7: Relative mass balance of alkaloids (caffeine and theobromine) at the extraction, filtration and adsorption / desorption stages. The percentage balance is calculated on the basis of the cocoa nibs, assuming that the cocoa nibs are 100%.

[0120]

[0121] The results show that, while theobromine is mainly present in the corresponding concentrated extract, caffeine is almost equally distributed between the theobromine-rich extract and the polyphenol concentrate. The concentration of theobromine in the theobromine-rich extract was determined to be 4.5 g / 100 g dry defatted matter.

[0122] Table 8 shows the content and distribution of cocoa flavanols (DP1-DP7) in the whole adsorption / desorption process with respect to the content of the permeate obtained in the filtration step. The results show that 75% of the flavanol content in the filtrate can be recovered in the polyphenol concentrate, while reducing the proportion of high degree of polymerization oligomers (DP6 and DP7) that are mainly present in the theobromine-rich extract. A favourable high content of monomeric flavanols (DP1) of 10.5 g / 100 g dry defatted matter is achieved, equivalent to a content of 23.4% by weight based on the total content of flavanols with a degree of polymerization from 1 to 7 (DP1 to DP7).

[0123] Table 8: Content distribution and yield of cocoa flavanols (DP1-DP7) after adsorption / desorption, determined by HPLC / fluorescence detection, with respect to the content in the hypothetical permeate (filtrate) taken as 100%.

[0124]

[0125] Table 9 summarises the content of cocoa flavanols in dry defatted matter for each intermediate product and final product. The results show that the process of the application provides a polyphenol extract in which the concentration of cocoa flavanols is advantageously as high as 44.8 g / 100 g dry defatted matter.

[0126] Table 9: Total content of cocoa flavanols (DP1-DP7) determined by HPLC / fluorescence detection.

[0127]

[0128] The total yield of cocoa flavanols in the polyphenol concentrate with respect to the content of cocoa flavanols present in the raw material (i.e. cocoa nibs) is 20.3%. Thus, it has been demonstrated that the process of the application is capable of simultaneously providing an extract with a high concentration of flavanols, a high proportion of monomeric and dimeric flavanols, and a very high yield of total polyphenols based on the total polyphenol content in the ingredients of the cocoa fruit.

[0129] To further characterize the extract obtained by this process, the relative content of nutrients (carbohydrates, dietary fibers, proteins) was determined in the context of the "8 categories of core nutrients analysis", including the calculation of the caloric value and the carbohydrate content (in compliance with the Regulation (EU) No 1169 / 2011), and the determination of the total fat content (by Weibull-Stoldt according to the ASU L44.00-4, 1985 standard), the saturated fatty acids content (by GC-FID method according to DGF C-VI 1985, modified according to DGF C-VI 1 le (98)), the dietary fiber content (by enzymatic-gravimetric method according to the ASU L00.00-18, 1997-01 standard), the protein content (by Kjeldahl titration), the water / dry mass content (by gravimetric method according to the ASU L44.00-4, 1985 standard), the total ash content (by gravimetric method), the sodium content (by flame AAS) and the sugar content (by HPLC-RI). The results of carbohydrates, dietary fibers and proteins are shown in Table 10, together with the content of the intermediate product of the filtration step.

[0130] Table 10: Relative content of carbohydrates, dietary fibers, proteins in the intermediate product and in the final product.

[0131]

[0132] The results show that the majority of the nutrients in the permeate are separated by the non-adsorbed fraction, providing an extract rich in theobromine. In the polyphenol concentrate, when considering the content in relation to the defatted dry matter, it is worth noting that the proteins constitute the largest part of the nutritional value, 40.1 g / 100 g of defatted dry matter, followed by the carbohydrates, 4.7 g / 100 g of defatted dry matter, and the dietary fibers, 1.5 g / 100 g of defatted dry matter.

[0133] Example 2

[0134] In example 2, 3387 kg of fermented, pasteurized and non-roasted cocoa nibs (origin: Peru) were subjected to the same extraction process as in example 1, with the difference that the hydrophilic extract was filtered at a temperature of 10 ± 2°C and under a filtration area of 30 m2of dynamic cross-flow filtration system using a ceramic membrane filter disc with a membrane pore size of 0.1 pm. In addition, contrary to example 1, in example 2, the P / DF-rich extract and the non-adsorbed permeate were not processed into a powder. 2

[0135] Table 11: Mass balance of cocoa butter throughout the extraction process. The calculation of the mass balance percentage is based on a percentage of 100% of cocoa nibs.

[0136] ​

[0137] The production of the biologically active alkaloids theobromine and caffeine was monitored by HPLC and the results are shown in Table 12.

[0138] Table 12: Mass balance of alkaloids (caffeine and theobromine) during extraction, filtration and adsorption / desorption stages. The percentage balance was calculated on the basis of cocoa nibs, assuming cocoa nibs to be 100% (n.m. = not measured).

[0139]

[0140]

[0141] Table 13 shows the mass balance of the cocoa flavanols (DP1-DP7) during the entire extraction, filtration and adsorption / desorption steps as determined by HPLC.

[0142] The study shows that the adsorption / desorption process enables a high yield of flavanols DP1-DP4 in the polyphenol powder, of 48% to 60% relative to the filtrate. For DP5 or higher flavanols, the yield is even higher, of 59% to 68%. Overall, i.e. across DP1-DP7, a yield of 55% is achieved.

[0143] The yield of flavanols (DP1-DP7) was calculated to be 20% across all process steps (extraction, filtration and adsorption / desorption).

[0144] Table 13: Mass balance of cocoa flavanols (DP1-DP7) during extraction and filtration as determined by HPLC / fluorescence detection (n.m. = not measured).

[0145]

[0146] Table 14 summarizes the content of cocoa flavanols in the defatted dry matter of the various intermediate products and final product. The results show that the process of the present application provides a polyphenol powder having a favourable high concentration of cocoa flavanols of 44.4 g / 100 g defatted dry matter. Advantageously, the polyphenol powder shows a monomeric flavanol (DP1) content of 9.70 g / 100 g defatted dry matter.

[0147] Table 14: Total content of cocoa flavanols (DP1-DP7) as determined by HPLC / fluorescence detection.

[0148]

[0149] Example 3

[0150] In Example 3, the same extraction procedure and analysis as in Example 2 was performed on 755 kg (part) of defatted, washed treated cocoa. As shown in Table 15, in contrast to Examples 1 and 2, no cocoa butter was recovered by hydrophobic liquid phase.

[0151] Table 15: Mass balance of cocoa butter during the entire extraction and filtration process. The calculation of the percentage of mass balance is based on 100% of cocoa.

[0152]

[0153] The production of the biologically active alkaloids theobromine and caffeine was monitored by HPLC and the results are shown in Table 16. In contrast to Examples 1 and 2, the yield of both caffeine and theobromine was higher during filtration and adsorption / desorption.

[0154] Table 16: Mass balance of alkaloids (caffeine and theobromine) during the extraction, filtration and adsorption / desorption stages. The calculation of the percentage of balance is based on the starting material, which is assumed to be 100% (n.m. = not measured).

[0155]

[0156]

[0157] Table 17 shows the mass balance of the cocoa flavanols (DP1-DP7) during the entire extraction, filtration and adsorption / desorption steps, as determined by HPLC.

[0158] In this experiment, 53% of the flavanols (DP1-DP7) were extracted into the hydrophilic liquid phase, while the remaining amount was retained in the cocoa powder. The low degree of polymerization flavanols (DP1-DP3) were efficiently extracted by the solvent, while the flavanols DP4-DP7 were mainly present in the cocoa powder. During the filtration process, about 48% to 72% of the initial content of flavanols DP1-DP4 was recovered in the permeate. For the flavanols DP5-DP7, the yield in the permeate was calculated to be 11% to 33%.

[0159] Table 17: Mass balance of cocoa flavanols (DP1-DP7) during the extraction and filtration process, as determined by HPLC / fluorescence detection (n.m. = not measured).

[0160]

[0161] Table 18 summarizes the content of cocoa flavanols in the defatted dry matter of the various intermediate products and final product of Example 3. As the results show, extraction led to an enrichment of flavanols from 15.8 g / 100 g ffTS in defatted cocoa in the cocoa extract to 20.04 g / 100 g ffTS in the dry hydrophilic phase. After filtration, flavanols were further enriched in the permeate. Finally, a polyphenol powder with an extremely high concentration of cocoa flavanols was obtained, of 88.2 g / 100 g defatted dry matter. These results show that the use of unfermented cocoa beans as raw material and / or the efficient separation of cocoa butter (which can negatively affect the adsorption efficiency) has a positive impact on both the yield of flavanols in the overall process step and the enrichment of flavanols in the final polyphenol powder.

[0162] Table 18: Total content of cocoa flavanols (DP1-DP7) determined by HPLC / fluorescence detection.

[0163]

[0164] Upon further examination of the flavanol (DP1-DP7) concentration at each stage of the diafiltration (15% initial cocoa extract content for each stage), it was observed that diafiltration significantly improved the yield of flavanols in the permeate.

[0165] Many other features, modifications, and improvements will occur to those skilled in the art once given this disclosure.

Claims

1. A process for obtaining an extract from a cocoa fruit ingredient, comprising the following steps: a) wet-milling the cocoa fruit ingredient in a polar solvent and separating a liquid phase from the wet-milling mixture to obtain a hydrophilic extract; b) optionally separating residual solids from the hydrophilic extract; c) adsorbing the hydrophilic extract on a non-ionic macroporous resin; d) desorbing the hydrophilic extract from the non-ionic macroporous resin with a water-miscible organic solvent to obtain a polyphenol extract; and e) concentrating the polyphenol extract to obtain a polyphenol concentrate.

2. The process according to claim 1, wherein step a) comprises the following steps: a1 ) adding a polar solvent to the cocoa fruit ingredient to form a suspension; a2) wet-milling the suspension; a3) heat-treating the suspension at a temperature of 70 °C or less; and a4) separating the suspension into a hydrophilic liquid phase (heavy phase), a hydrophobic liquid phase (light phase) and a solid phase, the hydrophilic liquid phase comprising the hydrophilic extract as a main component and residual solids as a minor component, the hydrophobic liquid phase comprising cocoa butter as a main component and solids and / or hydrophilic solvent as a minor component, and the solid phase comprising cocoa powder and hydrophilic solvent.

3. The process according to claim 1 or 2, wherein the polar solvent is selected from water, Ci-C8alcohols, C2-C8ketones, C3-C7esters, C2-C8ethers, C4-C 10 lactates and mixtures thereof, and can optionally include an acid, preferably acetic acid or citric acid; and / or, wherein the water-miscible organic solvent is selected from the group consisting of Ci-C8 alcohols, C3-C5 ketones, C3-C5 esters, C2-C4 ethers, C2-C5 aldehydes and mixtures thereof, preferably from the group consisting of methanol, ethanol, isopropanol or acetone.

4. The process according to any one of claims 1 to 3, wherein the cocoa fruit ingredient in step a) comprises unfermented and unbaked cocoa beans, preferably unfermented and unbaked cocoa beans which have not been dried.

5. The process according to any one of claims 1 to 4, wherein in step a) the cocoa fruit ingredient is subjected to a single or multiple wet-milling step to an average particle size of 100 pm or less, preferably 50 pm or less, more preferably 20 pm or less.

6. The process according to any one of claims 1 to 5, wherein the cocoa fruit ingredient and its extracts are not subjected to a temperature higher than 70 °C.

7. The process according to any one of claims 1 to 6, wherein in step e) the polyphenol concentrate is subjected to a concentration or evaporation step and optionally a spray-drying step to obtain a polyphenol powder.

8. The method of any one of claims 1 to 7, wherein the non-ionic macroporous resin has a surface area of at least 400 m2 / g as determined by nitrogen BET method and / or a total pore volume of at least 0.90 cc / g as determined by nitrogen BET method. 2 / g of surface area as determined by nitrogen BET method and / or a total pore volume of at least 0.90 cc / g as determined by nitrogen BET method.

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

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

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

12. Polyphenol extract obtained by the method according to any one of claims 1 to 11, preferably fulfilling at least one of the following characteristics: a total flavanol content of at least 25 g / 100 g dry defatted matter, preferably at least 30 g / 100 g dry defatted matter, further preferably at least 50 g / 100 g dry defatted matter, particularly preferably at least 53 g / 100 g dry defatted matter, as determined by HPLC; a content of monomeric flavanol (DP1) of at least 18 wt.-%, preferably at least 20 wt.-%, more preferably at least 24 wt.-%, based on the total content of flavanols with a degree of polymerization of 1 to 7 (DP1 to DP7), as determined by HPLC; and a protein content of less than 42 g / 100 g dry defatted matter, a carbohydrate content of less than 6 g / 100 g dry defatted matter, and a dietary fiber content of less than 2 g / 100 g dry defatted matter.

13. Protein-enriched extract obtained by the method according to claim 10.

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

15. Food composition, food supplement, pharmaceutical composition or cosmetic composition comprising the extract according to claims 12 to 14.

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