CN101506190A - Products containing polyphenols - Google Patents

Abstract

This invention discloses shelf-stable products, particularly beverages, containing polyphenols and optionally sterol esters and/or steranol esters. Loss of polyphenols during heat treatment is prevented by lowering the product's pH by at least 0.2 after mixing and before heat treatment. The final beverage should have a pH of about 4.6 to about 6.8. Preferred products are low-fat cocoa beverages containing at least 0.2 mg to 5 mg of cocoa polyphenols per gram of beverage.

Description

Products Containing Polyphenols

Cross References to Related Applications

This application is a claim of Provisional Application Serial No. 60/695,361, filed June 29, 2005, entitled "Formulations for Vascular Health" and filed June 29, 2005, entitled "Process for control theIsomerization of(-)-Epicatechin and(+)-Catechin in Edible Food Products PCT application claiming priority to Serial No. 11/170,593.

field of invention

The present invention relates to products, especially beverages, containing polyphenols and optionally sterol esters and/or stanol esters, and to a process for the production of said products. Products prepared by the method of the invention contain preserved levels of polyphenols.

Background of the invention

Polyphenolic compounds are biologically active substances obtained from plant materials. They are closely related to the organoleptic and nutritional qualities of products obtained from plant materials. Many plant polyphenols have antioxidant activity. Consumption of polyphenols present in tea, red wine and cocoa provides significant health benefits.

Standard sterilization process techniques for the preparation of ready-to-eat or ready-to-drink products result in the loss of polyphenols such as flavan-3-ols and proanthocyanidins.

Therefore, it would be highly desirable to prepare ready-to-eat or ready-to-drink products, especially milk drinks, in which the polyphenols are not lost but preserved during the heat treatment required to prepare the final product and produce a shelf-stable product. Heat treatment of milk beverages at neutral or near-neutral pH is of particular concern because the time required to sterilize the beverage can cause denaturation of milk proteins.

Summary of the invention

A heat-processed ready-to-eat or ready-to-drink product is prepared that contains at least about 0.2 milligrams of polyphenols per gram of product. The amount of polyphenols is preferably from about 0.2 to about 5. The product has a moisture content of at least about 5% by weight. When the product is a beverage, the moisture content is from about 50% to about 80%. The pH of the final product is from about 4.8 to about 6.8, preferably from about 6.2 to about 6.8. Polyphenols are food flavan-3-ols, such as catechin, epicatechin, gallocatechin, epigallocatechin, and/or afurcatechin; and/or food proanthocyanidins , for example proanthocyanidins, prodelphinidins and/or propelargonidins.

Preferred products include milk cocoa beverages, soy cocoa beverages and whey cocoa beverages containing cocoa polyphenols such as (±)-catechin, (±)-epicatechin, and/or proanthocyanidin oligomers thereof. One or more high CP cocoa ingredients including part-skim cocoa powder, full-skim cocoa powder, chocolate liquor, liquid cocoa extract and/or dry cocoa extract are used in the cocoa beverage.

Partially defatted cocoa powders contain at least about 25 mg, preferably about 25 mg to 35 mg, of cocoa polyphenols per gram of said defatted cocoa powder. The chocolate mass contains about 12 mg, preferably about 13 mg to 17 mg of cocoa polyphenols per gram of said skim mass. The cocoa extract contains at least about 200 mg, preferably about 350-500 mg of cocoa polyphenols per gram of dry extract.

Adjust the pH with a food acid such as citric acid or a food base such as sodium hydroxide.

An improved process for the preparation of a ready-to-eat or ready-to-drink product containing polyphenols comprises the step of lowering the pH of the product by at least 0.2, preferably 0.4 after mixing and before heat treatment. Typically, the pH prior to heat treatment is below about 7.5, preferably from about 4.6 to about 6.8.

A method for preparing a ready-to-drink cocoa beverage containing one or more proanthocyanidins comprising the steps of: adding one or more proanthocyanidins to the beverage; lowering the pH of the beverage by at least 0.2, to about 4.6 to about 6.8, with an edible acid ; heat-treating a pH-adjusted beverage; and packaging the heat-treated beverage. The heat treatment is performed at about 145°F to about 280°F for about 1 second to about 15 seconds.

A method for preparing a cocoa beverage containing at least about 0.2 milligrams of cocoa polyphenols per gram of beverage comprises the steps of: under high shear, water and a dry cocoa mixture comprising one or more cocoa ingredients; A dry blend of a sweetener, one or more thickeners, one or more stabilizers, and a vitamin-mineral mixture is mixed into milk; cocoa syrup is added to the milk mix; and liquid is added for consumption spices. Use food acid or food base to adjust the pH from about 6.8 to about 7.5, preferably from about 6.8 to about 7.2, more preferably from about 6.9-7 to about 4.6-6.8, preferably from about 6.2 to about 6.8. The slurry is heat treated at about 161°F to about 280°F for about 1 second to about 15 seconds. The slurry is optionally cooled and/or homogenized prior to packaging. Typically, the beverage has a water content of about 50%. Cocoa ingredients are preferably prepared from unfermented and/or underfermented cocoa beans. Cocoa extract is prepared by solvent extraction of defatted, unfermented or underfermented cocoa beans and removal of the solvent. The beverage also contains alkalized cocoa powder, an emulsifier such as lecithin, a sweetener such as sugar, one or more thickeners, one or more stabilizers, a vitamin-mineral mixture, and /or one or more flavorings. (-)-epicatechin and (+)-catechin at a water content of at least about 50% by weight by lowering the pH of the product by at least 0.2, typically to about 4.6 to about 6.8, prior to heat treatment The loss of polyphenol-containing products during heat treatment is minimized. Cocoa polyphenols present in beverages after heat treatment include (+)-epicatechin, (-)-epicatechin, (+)-catechin, (-)-catechin and proanthocyanidin di polymers and their trimers.

Preferably, the cocoa beverage contains heart healthy vitamins such as B6 and B12, antioxidants such as vitamin E and vitamin C, and/or minerals such as calcium. In a preferred embodiment, the product contains phytosterol esters and/or stanol esters. The amount of sterol ester and/or stanol ester is about 0.1 mg to about 30 mg per gram of product. For maximum benefit, the beverage should be used as part of a diet low in saturated fat and cholesterol.

Brief description of the drawings

Fig. 1 is a schematic diagram showing how to heat treat a product. Conventional ingredients are combined, mixed and pH adjusted prior to terminal sterilization and packaging processes. S1 to S5 are sampling points whose total cocoa polyphenol (CP) content is measured during the production process.

Fig. 2 is a schematic diagram describing the sterilization process of a typical high CP milk beverage. The high CP cocoa powder, dried high CP cocoa extract and alkalized cocoa powder were subjected to primary slurrying under high shear at 60°C (180°F) to hydrate the cocoa and completely disperse all dry ingredients with the cocoa powder. The pH of the mixture is adjusted, preferably below 6.5, by adding citric acid. It is important to note that the pH adjustment of the completely untreated batch is done prior to heat treatment. The beverage is then sterilized and packaged.

Figure 3 is a normal phase HPLC/FLD spectrum of high CP cocoa powder.

Figure 4 is a normal phase HPLC/FLD profile of high CP cocoa extract.

Figure 5 is a normal phase HPLC/FLD profile of cooked high CP cocoa extract.

Figures 6A and 6B are normal phase HPLC/FLD profiles of high CP cocoa beverages before and after UHT heat treatment.

Detailed description of the invention

A "foodstuff" as used herein is a protein, carbohydrate and/or fat containing material that is used in any living organism to maintain growth, repair vital processes and provide energy. Foodstuffs may also contain supplementary substances such as minerals, vitamins and flavorings (Merriam-Webster Collegiate Dictionary, Tenth Edition, 1993). The term "foodstuff" includes beverages suitable for human or animal consumption. "Food additive" is defined by FDA in 21 C.F.R. 170.3(e)(1) and includes both direct and indirect additives.

As used herein, a "dietary supplement" is a product (other than tobacco) that is intended to supplement the diet and has or contains one or more of the following dietary ingredients: vitamins, minerals, herbs or other botanicals, amino acids, Daily Intake Dietary substance intended to supplement the diet, or a concentrate, metabolite, component, extract or combination of these ingredients. (Merriam-Webster Collegiate Dictionary, Tenth Edition, 1993). When the term is used on food labels, "supplement" refers to a nutrient added in an amount exceeding 50% of the recommended U.S. daily intake (“Understanding Normal and Clinical Nutrition,” 3rd ed., Editors Whitney, Catalado, , p. 525).

The present invention relates to aqueous products, especially beverages, containing flavan-3-ols such as catechin, epicatechin, gallocatechin, epigallocatechin and/or afurcatechin and proanthocyanidins such as procyanidins, prodelphinoids and progeranium pigments, said products are used in a preferred embodiment in combination with sterol ester(s) and/or stanol ester(s) .

Cholesterol-lowering agents other than sterol/stanol esters may be used herein in combination with sterol ester(s) and/or stanol ester(s). Examples include low-calorie and no-calorie fats.

The product may contain L-arginine, minerals such as calcium, potassium, and magnesium, vitamins such as vitamins B, C, and E, carotenoids, and mono- or polyunsaturated fatty acids (eg, omega-3 fatty acids). Polyphenols derived from sources other than cocoa may also be used alone or in combination with cocoa proanthocyanidins, said polyphenols having antioxidant properties similar to these cocoa proanthocyanidins. These sources include nuts, nut flours and nut skins.

As used herein, the term "cocoa polyphenols" refers to polyphenolic compounds present in cocoa beans, cocoa nibs, and cocoa ingredients prepared from cocoa beans or cocoa nibs, such as partially or fully defatted cocoa powder, chocolate mass, and liquid or dry cocoa extract.

The term "procyanidins" refers to naturally occurring or synthetically derived oligomers of catechins and/or epicatechins, however, any reference herein to "cocoa procyanidins" should be understood to also include (±)-catechins Monomer of theanin and (±)-epicatechin. (+)-catechin, (-)-epicatechin and their respective epimers (-)-catechin and (+)-epicatechin have the following structures:

Proanthocyanidin oligomers may contain from 2 to about 18 monomeric units. The oligomers include, for example, dimers, trimers, tetramers, pentamers, hexamers, heptamers, octamers, nonamers, decamers, and the like. In natural oligomers, the monomers are linked by (4→6) and/or (4→8) interflavan linkages. Oligomers with only (4→8) linkages are linear. The presence of at least one (4→6) linkage results in branched oligomers. For the synthesis of (4→8) proanthocyanidins, refer to US Patent 6,420,572, the contents of which are incorporated herein by reference. The '572 patent discloses phenolic monomers (e.g., epi catechin or catechin) is coupled with another protected phenolic monomer to produce a protected dimer which is subsequently deprotected, or optionally combined with other protected, activated phenolic Monomer coupling.

Cocoa polyphenol derivatives are also useful herein. They include monomers and oligomers of gallated catechin and/or gallated epicatechin, glycosylated monomers and oligomers, and mixtures thereof; metabolites of proanthocyanidin monomers and oligomers , such as sulfated forms, glucuronidated forms, and methylated forms; and enzyme cleavage products of proanthocyanidins produced by colonic microorganisms from metabolism or internal mammalian metabolism (internal mammalian metabolism). The derivatives may be from natural sources or prepared synthetically.

Synthetic oligomers are also useful herein. See US Patent 6,156,912 to W. Tückmantel et al., issued December 5, 2000 and US Patent 6,476,241 to A. Kozikowski et al., issued November 5, 2002.

The term "medium quality cocoa beans" refers to cocoa beans that have been separated from the pulp material and dried. It is relatively free from mold and infection. Such beans are commercially available and used as raw material for the preparation of high CP cocoa solids and cocoa powder, high CP chocolate liquor and high CP cocoa extract. The term includes any genetically modified or produced such beans.

The term "freshly harvested raw cocoa beans" refers to freshly harvested cocoa beans (also referred to as seeds) from cocoa pods that have not been processed except for separation from the pulp. Cocoa beans from any species of Theobroma, Herrania, or hybrids thereof within and between species can be used to prepare the cocoa ingredients used herein, ie, cocoa powder, chocolate liquor, and cocoa extract. Preferably, said cocoa ingredient is prepared from unfermented and/or underfermented cocoa beans having a fermentation factor below 275. "Fermentation factor" is determined using an industry-accepted rating system. To evaluate the degree of fermentation, cocoa beans are usually subjected to a standard cut test to evaluate the quality defined by the standard. Blue-gray beans, purple-red beans, a mixture of blue-gray and purple-red beans, a mixture of purple-red and brown beans, or a mixture of blue-gray, purple-red, and brown beans can be used. More preferably, the cocoa beans are blue-gray and/or purple-red cocoa beans because they have a higher cocoa polyphenol content than fermented beans.

cocoa toppings

As used herein, the term "cocoa powder" refers to partially or fully defatted cocoa powder (e.g., blocks or solids) prepared directly by extruding shelled cocoa beans (e.g., by screw extrusion) into cocoa fat and partially defatted cocoa solids, or by grinding roasted cocoa beans into chocolate mass and pressing the chocolate mass to recover cocoa butter and partially defatted cocoa solids.

As used herein, the term "high CP cocoa ingredient" refers to a cocoa ingredient that preserves flavan-3-ols and proanthocyanidins during the preparation of the cocoa ingredient, and/or refers to a cocoa ingredient prepared from unfermented or underfermented cocoa beans . The high CP cocoa powder contains at least about 25 milligrams, preferably 25 milligrams to 50 milligrams, most preferably 25 milligrams to 35 milligrams of cocoa polyphenols per gram of skim powder. The high CP chocolate mass contains at least about 10 mg, preferably about 12 mg to about 25 mg, most preferably 13 mg to about 17 mg of cocoa polyphenols per gram of skim pulp. The high CP cocoa extract contains at least 200 mg, preferably 250 mg to 500 mg, most preferably about 350 to about 500 mg of cocoa polyphenols per gram of dry extract.

Kirk S. Kealey et al., US Pat. No. 6,015,913 issued Jan. 18, 2000, discloses a process for preparing cocoa solids with high cocoa polyphenol content directly from cocoa beans, the contents of which are incorporated herein by reference. In the method of the '913 patent, the cocoa beans are heated for a period of time at an internal bean temperature (e.g., infrared heating to about 100° C. to about 110° C.) sufficient to loosen the cocoa shell without roasting the cocoa bean flesh. The '913 patent describes a method for determining internal bean temperature (IBT). Wind-picked cocoa beans from the cocoa shell. The cocoa nibs are extruded into cocoa butter and partially defatted cocoa solids. Cocoa solids contain cocoa polyphenols including cocoa proanthocyanidins from cocoa beans. The amount of higher procyanidin oligomers present in the cocoa solids thus produced is higher than the amount of higher procyanidin oligomers present in cocoa solids produced by conventional roasting processes. The total cocoa proanthocyanidin content of cocoa powder and cocoa extracts was determined as described below.

Kirk S. Kealey et al. in U.S. Patent 6,312,753 issued November 6, 2001 disclose a method for preparing cocoa solids or chocolate liquor from roasted unfermented, underfermented, or medium to average quality cocoa beans, Its content is incorporated herein by reference. Use cocoa beans or cocoa beans with a fermentation factor of 275 or less. The beans are passed through infrared heaters and winnowed to separate the shells (husks), roasted to an internal bean temperature of about 95°C to about 150°C, and ground into a coarse chocolate mass from which cocoa butter and part of the Defatted cocoa solids.

Preferably, the products herein, such as cocoa beverages, comprise cocoa extract to increase the total cocoa polyphenol content of the product. Leo J. Romanczyk, Jr. et al. disclose a method of preparing cocoa extract from cocoa beans in US Patent 5,554,645, issued September 10, 1996, the contents of which are incorporated herein by reference. The method of the '645 patent includes: freeze-drying cocoa beans containing pulp; depulping the freeze-dried mass; dehulling and grinding the freeze-dried cocoa beans; and defatting the resulting cocoa mass followed by solvent extraction, the Solvents such as aqueous methanol, aqueous acetone or ethyl acetate. Extracts can also be prepared from high CP cocoa solids prepared by the methods described in the '913 patent and the '753 patent discussed above. U.S. Patent 6,627,232 issued September 30, 2003 to John F. Hammerstone et al. describes a process for the preparation of cocoa extracts enriched in certain oligomers and decaffeinated and detheobrominated cocoa extracts , the contents of which are incorporated herein by reference. Tetramers and higher molecular weight cocoa proanthocyanidin oligomers can be obtained by extracting high CP cocoa solids prepared from non-roasted cocoa beans with ethyl acetate and re-extracting the extracted solids with acetone, ethanol or a mixture of up to A mixture of 50% water is extracted. The ethyl acetate extract was enriched in monomers, dimers and trimers.

Alkalized cocoa powder is added to improve the flavor and color of the drink. The FDA Standard of Identity for alkalized cocoa allows the use of up to 3% anhydrous potassium carbonate or its alkaline equivalent, based on the weight of the roasted pulp.

Sterol Esters and/or Stanol Esters

Phytosterols are phytosterols that are insoluble in water and have a molecular weight and structure similar to cholesterol. More than forty phytosterols have been identified, with β-sitosterol, campesterol and stigmasterol being the most abundant. Other examples of useful sterols are brassicasterol, 24-dehydrocholesterol, chalinosterol, poroserol, and spongesterol.

Stanols are saturated derivatives of sterols in which all carbon-carbon bonds in the ring are saturated. Stanols generally contain 28 or 29 carbon atoms and include beta-sitostanol, spongostanol, 22,23-dyhydrobrassicastanol, and campestenol. Stanols are found in nature in small amounts, but stanols can be readily prepared from sterols by hydrogenation of sterols using any of a variety of methods known to those skilled in the art. When the sterol starting material is prepared from plant material, it will contain a mixture of different sterols. Therefore, the resulting stanols after hydrogenation will also be a mixture of different stanols.

The esterified forms of sterols and stanols are the preferred forms for use herein. Esterification renders sterols/stanols more soluble in fats and oils. For example, sterols can be esterified with fatty acid esters of oils such as rapeseed oil, canola oil, and the like. Suitable fatty acids include saturated or unsaturated fatty acids generally containing 14-24 carbon atoms. Examples of esterified sterols include sitosterol acetate, sitosterol oleate, and stigmasterol oleate. Stanol esters can be prepared as known in the art, for example, as described in: U.S. Patent 6,174,560 issued January 16, 2001 to Miettenen et al. and assigned to Raisio Benecol Ltd.; issued February 29, 2000 US Patent 6,031,118 issued to van Amerongen et al. and assigned to Lipton; US Patent 5,958,913 issued September 28, 1999 to Miettenen et al. and assigned to Raisio Benecol; issued April 6, 1999 to Higgins, III and assigned US Patent 5,892,068 to McNeil PPC, Inc.; and US Patent 5,502,045 issued March 26, 1996 to Miettenen et al. and assigned to Raisio Benecol, Ltd, the contents of which are incorporated herein by reference. The '045 patent describes the transesterification of a mixture of methyl esters of _C2 - _C22 fatty acids (eg, rapeseed oil) with free stanols using a transesterification catalyst such as sodium ethoxide. Transesterification methods such as those disclosed in the '045 patent can also be used to esterify sterols. In another embodiment, useful stanol esters are prepared by esterifying at least one sterol with a _C2 - _C22 fatty acid ester as described in the above-cited '913 patent.

Particularly useful herein are canola sterol, sunflower sterol and mixtures thereof. These sterol ester mixtures melt at around 30°C-50°C; however, typically the esters are heated to about 60°C-80°C to ensure that the entire mixture is liquefied. The liquid or liquefied (ie melted) sterol/stanol esters and emulsifiers are mixed with the cocoa slurry either before or after the addition of milk.

Emulsifier

Preferably, an emulsifier such as lecithin, mono- or di-glycerides is added to the cocoa premixed with the sterol esters and/or stanol esters or separately with the sterol esters and/or stanol esters , mono- or diglycerides, phospholipids, monoglycerides and esters of acetic, lactic, citric, succinic or tartaric acids, fatty acid esters of polyglycerol, sorbitol esters, sucrose esters, propylene glycol or polyglycerol polyresorcinoleate. The emulsifier is used in an amount of about 0.05% to about 5%, preferably about 0.05% to about 1%, more preferably about 0.05% to about 0.25%. Emulsifiers are well known to play a key role in the rheology of suspensions and are widely used in food production, especially confectionary and chocolate production, to enhance the rheology of solids suspensions (i.e. reduce viscosity and/or yield value) .

Lecithins obtained from vegetable oils such as soybean oil, cottonseed oil, corn oil, safflower oil, and rapeseed oil, including clarified lecithin, fluidized lecithin, compounded lecithin, hydroxylated Lecithin from ®, deoiled lecithin, and fractionated lecithin are useful herein. Soy lecithin is a preferred emulsifier for use herein. It is one of the oldest and most widely used emulsifiers. Based on the finished product, it can be used in an amount up to about 5% by weight, preferably about 0.05% by weight to about 0.3% by weight, more preferably about 0.1% by weight to about 0.3% by weight. The pH can be adjusted downward by 50% aqueous citric acid (by weight) or upward by 50% aqueous sodium hydroxide (by weight).

sweetener

Food carbohydrate sweetener(s) and/or sugar substitute(s) are used herein. Suitable sweeteners include those commonly used in food products and include, but are not limited to, sucrose (e.g., from sugar cane or sugar beets), glucose, fructose, lactose, maltose, glucose syrup solids, corn syrup solids, invert sugar, Invert sugar, honey, maple sugar, brown sugar, molasses, etc. Suitable sugar substitutes can be used to partially replace the food carbohydrate sweeteners. The term "sugar substitute" includes high-potency sweeteners, sugar alcohols (polyols), and bulking agents, or combinations thereof. High-potency sweeteners include aspartame, cyclamate, saccharin, acesulfame, neohesperidin dihydrochalcone, sucralose, thiamin, and stevia sweeteners (stevia sweetener), glycyrrhizic acid, African sweetener, etc., and mixtures thereof. Preferred high potency sweeteners include aspartame, cyclamate, saccharin and acesulfame-K. Examples of sugar alcohols may be any commonly used in the art, including sorbitol, mannitol, xylitol, maltitol, isomalt, lactitol, and the like. The food products of the present invention may also contain bulking agents, usually in combination with high-potency sweeteners. The term "filler" defined herein can be any commonly used in the art, including polydextrose, cellulose and its derivatives, maltodextrin, gum arabic, and the like.

other ingredients

Other water-soluble or water-dispersible ingredients also comprise lesser amounts in the beverage, for example up to about 35% thickening agent, preferably about 0.01-about 5%; Up to __% stabilizer, preferably about 0.2%-about 3% %; up to 1.0% vitamins and/or minerals, preferably from about 0.01% to about 0.35%; up to 1% flavoring; and up to 0.5% salt.

stabilizer

Phosphates are used as protein stabilizers. They protect proteins, such as those in milk, from granulation due to heat and acid shock. Milk proteins are more stable at neutral pH under conditions of intense heating, such as those used in sterilization processes.

Another method of stabilizing proteins is a thermal pretreatment step. Preheat the liquid containing the protein to around 160℉-180℉ to increase the stability of the protein to heat shock during sterilization. The pretreatment step unfolds and partially denatures the whey proteins, thereby reducing the possibility of granulation during the sterilization process.

Gums such as carrageenan are often used at ppm levels to suspend the cocoa solids in the milk drink. Microcrystalline cellulose is also used in cocoa beverages to enhance mouthfeel. It works synergistically with carrageenan to suspend cocoa powder.

As used herein, the term "flavorant" refers to a perfuming compound or composition used in foods and beverages to impart a desired taste and/or aroma. Exemplary flavorants suitable for use herein include vanillin, chocolate, fruit such as blueberry, raspberry, strawberry, and banana, spices, and naturally expressed citrus or sesame oils.

heat treatment

The product can be heat treated by the following methods: tank pasteurization, which requires heating at about 63°C (145°F) for about 30 minutes; high temperature short-time pasteurization (HTST), which requires heating at about 89°C (191°F). ) for about 1.0 seconds; or Ultra High Temperature (UHT), which requires heating at about 138°C (280°F) for about 2 seconds.

DH regulation

In most products, the pH is lowered by at least 0.2, preferably about 0.4, most preferably 0.6. The product pH prior to heat treatment should be from about 6.8 to about 7.5, preferably from about 6.8 to about 7.3, or most preferably from about 6.9 to about 7.2. When the pH was adjusted from 7.2 to 6.5, the CP loss was reduced by up to 30%. When the pH was adjusted from 6.8 to 6.2, the retention of polyphenols increased by about 16%. Before heat treatment, the total loss is only about 20%. After heat treatment, the total loss increases to about 35%.

In addition to preserving cocoa polyphenols, pH adjustment provides a method for enhancing flavor and/or reducing bitterness and astringency associated with polyphenols.

Test Methods

The percent CP loss was calculated by subtracting the final CP amount from the starting CP amount and dividing by the starting CP amount and multiplying by 100.

Determination of the content of total cocoa proanthocyanidins

The total cocoa proanthocyanidin content of powders, cocoa extracts and beverages was determined by normal phase high performance liquid chromatography (HPLC) on silica gel with a fluorescence detector. Details of this method are found in Adamson et al., J. Ag. Food Chem. 47(10) 4184-4188 (1999). The cocoa solids were defatted with hexane prior to extraction of proanthocyanidins. The hexane was removed and the solid was extracted with acetone. A total of 32-35 g of cocoa beverage was quantitatively transferred to a 100 mL volumetric flask. Add 0.5 mL of glacial acetic acid to it and dilute the solution with acetone. This solution was compared to the extraction solvent used for all other samples, which consisted of 70:29.5:0.5 acetone:water:acetic acid [volume ratio]. Beverage samples were not defatted prior to analysis. The water of the beverage is used to prepare the aqueous portion of the extraction solvent. Quantification of cocoa proanthocyanidins was achieved by using a highly purified cocoa polyphenol extract. The samples were then compared as described in the paper by Adamson et al. to accurately determine the amount of catechin, epicatechin and proanthocyanidin oligomers.

The following steps were used to prepare and test high CP products.

sample preparation

In Example 1, the drink was freeze-dried and extracted twice with an acetone/water/acetic acid mixture (CH ₂ COCH ₃ :H ₂ O:HOAc, 79.5:20:0.51), sonicated at 50° C. for 15 minutes, centrifuged at 35,000 rpm 6 minute. The solvent was recovered from the collected supernatant under reduced pressure or vacuum and lyophilized. The resulting material was used for normal phase HPLC analysis. A total of 32-35 g of cocoa beverage was quantitatively transferred to a 100 mL volumetric flask. Add 0.5 mL of glacial acetic acid to it and dilute the solution with acetone.

In Example 3 and Example 4, the sample preparation method of cocoa drinks was improved to better adapt to the analysis of high-moisture products. Sample preparation consisted of taking 32-35 g of drink and quantitatively transferring it to a 100 mL volumetric flask; adding 0.5 mL of glacial acetic acid; and adding acetone. This method yielded a solvent that could be used for comparison with the extraction solvent of other cocoa samples consisting of 70:29.5:0.5 - acetone:water:acetic acid (volume ratio). Beverage samples were not defatted prior to analysis. The aqueous portion of the extraction solvent is prepared from the water of the beverage.

Normal phase chromatography-HPLC/MS analysis (method of Adamson et al.)

For Example 1, the normal phase HPLC method published by Adamson et al. (J. Agric. Food Chem., 1999, 47, pp. 4184-4186) was used.

The conditions are as follows:

Stationary phase: Hypersil ODS 100 x 4.6mm 5μm particle size.

Mobile phase A: 0.170 HOAc in water.

Mobile phase B: 0.170 HOAc in methanol.

Flow rate: 1.0mL/min

gradient:

time (minutes) %B 0 15 20 25 30 75 45 100

The column used was a 250 x 4.6-mm, id, 5 μm Developosil diol (Phenomenex, Torrance, CA). The binary mobile phase consisted of (A) CH ₃ CN:HOAc, (98:2, volume ratio) and (B) CH ₃ OH:H ₂ O:HOAc (95:3:2). Separation was carried out at 30°C with a flow rate of 1.0 mL/min according to the following linear gradient: 0-35 min, 0-40% B; 35-45 min, 40% B isocratic elution; 45-46 min, 40-0% B, Maintained at 0% B for 4min. The eluate was monitored with a fluorescence detector with an excitation wavelength of 276 nm and an emission wavelength of 316 nm.

The following examples are intended only as illustrations of certain preferred embodiments and not as limitations of the invention.

Example

Example 1: Preparation of high CP cocoa solids from cocoa beans

Commercially available cocoa beans having an initial moisture content of about 7% to 8% by weight are precleaned in a primary cleaning sieve. The pre-cleaned beans from the primary cleaning screen were further cleaned in an air fluidized bed density separator. The cleaned cocoa beans are then passed through an infrared heating unit at a rate of approximately 1,701 kilograms per hour. The depth of beans in the vibrating bed of the apparatus was about 2-3 beans deep. The surface temperature of the apparatus was set at about 165°C, thereby generating an internal bean temperature (IBT) of about 135°C in the range of 1 minute to 1.5 minutes. This treatment causes the shells to dry quickly and separate from the cocoa pulp. Before the winnowing step, the fragments separated by the vibrating screens preceding the unit are reintroduced into the product stream. The resulting beans after micronization should have a moisture content of about 3.9% by weight. The beans were exposed to an internal bean temperature of about 135°C and immediately cooled to about 90°C in about three minutes to minimize additional moisture loss. The beans are then winnowed to split the beans, loosen the shells and separate the lighter shells from the pulp while minimizing the loss of pulp using a shell reject stream. The resulting cocoa pulp was pressed using two screw presses to extract the fat from the cocoa solids.

A sample of cocoa solids produced from unfermented cocoa beans (fermentation factor 233) according to the above method will typically have a total cocoa proanthocyanidin content of from about 50 mg to about 75 mg, Preferably from about 60 mg to about 75 mg, or more preferably from about 75 mg to about 80 mg total cocoa proanthocyanidins. Figure 3 shows the normal phase HPLC profile of high CP cocoa powder.

Example 2: Preparation of Cocoa Extract

The cocoa solids from Example 1 are contacted with an aqueous organic solvent for 0.5 hours to 2.5 hours at room temperature. For Cocoa Extract A, the solvent was about 75% ethanol/25% water (by volume). For Cocoa Extract B, the solvent was about 80% acetone/20% water (by volume). The micelles are separated from the cocoa residue and concentrated by evaporation. The concentrated extract is then spray dried. The HPLC/FLD profile of cocoa extract is shown. Figure 4 shows the spectrum of the sample before heating. Figure 5 shows the chromatogram of the ethanol extract after heating and refluxing in deionized water overnight.

Example 3: Preparation of high CP cocoa beverages containing sterol esters

A low-fat cocoa drink prepared from: milk, high CP cocoa powder, high CP extract, alkalized cocoa powder, lecithin, canola, granulated sugar, blend of cellulose and carrageenan, lemon salt/phosphate stabilizer blend, vitamin-mineral premix, and chocolate and vanilla flavor. Use 1% milk. The prescriptions are listed in Table 1.

Table 1

ingredients % weight range water 10-40 low fat milk 50-90 granulated sugar 5-10 High CP cocoa powder 0.5-3 Alkalized cocoa powder 0.5-3 High CP Extract 0.5-3 Canola 0.2-1 Lecithin 0.30 Cellulose 0.35 carrageenan 0.01-0.3 Phosphate 01.-0.4 vitamin-mineral blend 0.1-1.0 cocoa spice 0.200 natural flavors 0.1-1.0 total 100.00

The cocoa ingredients were slurried in water at 180°F. Mix remaining dry ingredients with 1% milk and add to cocoa slurry. Adjust pH to 6.5-6.8 with citric acid. Process the cocoa drink at 280°F for 6.5 seconds and pack.

Figures 6A and 6B represent typical profiles of beverages before and after UHT treatment.

Example 4: Preparation of high CP cocoa beverages without sterol esters

Two beverages were prepared from high CP cocoa solids and high CP chocolate liquor that was not high CP extract. The ingredients used are listed in Table 2 below.

Table 2

ingredients Beverage A Beverage B 1% milk 79.77 86.37 granulated sugar 9.50 9.50 High CP cocoa powder 2.20 2.20 Alkalized cocoa powder 2.20 1.10 chocolate syrup 1.00 0.00 cocoa crumbs 4.00 0.00 malted milk powder 0.5 0.00 chocolate spice 0.20 0.20 vanilla spice 0.25 0.25 Cellulose 0.37 0.37 carrageenan 0.01 0.01 total 100.00 100.00

Surprisingly, despite the high level of cocoa polyphenols, the beverage was less bitter and astringent than expected.

Example 5: Effect of Process Variables

The effect of process variables on the loss of cocoa polyphenols (CPs) was investigated. The CP content of cocoa powder is 61.4mg/g defatted powder. The CP content of the extract was 384.99mg/g extract. The extract is a dry ethanolic aqueous extract prepared from unfermented cocoa beans as described in Example 2. Percent loss was calculated based on CP levels after treatment and before treatment. The results are summarized in Table 3.

table 3

Beverage No. CP level before treatment (mg/g) (S1/S4) CP level after treatment (mg/g) (S5) CP loss (%) CPsmg per serving Test purpose and notes 1 1.40 1.06 24.3 229 Test of the sterilization process for CP of water-in-oil milk protein. (pH=6.5) 2 1.40 0.97 30.7 210 Test of the sterilization process for CP containing dairy proteins. (pH=6.5) skimmed milk powder 3 1.40 0.71 49.3 153 The conditions are similar to No.2, alternatively pH=7.2 (conventional chocolate milk drink) - reconstituted milk 4 1.46 0.72 50.6 156 Recipe No.4pH6.8 5 1.39 0.91 34.5 197 Formula No.5pH6.2

The results showed that the loss of CPs occurred at pH 7.2 was 49.3%, while at pH 6.5 the loss was only 30.7%. In Beverage 4 and Beverage 5 (both based on the same formulation), reducing the pH from 6.8 to 6.2 reduced the loss of CPs from 50.6 to 34.5%. Figures 6A and 6B are typical normal phase HPLC profiles of beverages before and after UHT treatment.

Other variables and modifications apparent to those skilled in the art are within the scope and teaching of these inventions. The invention is not to be restricted except as set forth in the appended claims.

Claims (31)

1. A heat-treated ready-to-eat or ready-to-drink product, said product having a water content of at least about 5% by weight and a pH of about 4.8 to about 6.8, containing at least about 0.2 mg of polyphenols per gram of said product, wherein said product The polyphenols are edible flavan-3-ols and/or edible proanthocyanidins. 2. The product of claim 1 which is a beverage having a water content of about 50% by weight to about 80% by weight and a pH of about 6.8 to about 7.2, said product containing from about 0.2 mg to about 5 mg of polyphenols, wherein the flavan-3-ols are catechins, epicatechins, gallocatechins, epigallocatechins, and/or afurcatechins, and wherein the proanthocyanidins are proanthocyanidins , Protodelphinium pigments or protogeranium pigments. 3. The product of claim 1 having a water content greater than 80% by weight and a pH of from about 6.9 to about 7.0, and said product contains from about 0.2 milligrams to about 5 milligrams of polyphenols, wherein said flavan- The 3-alcohols are catechins, epicatechins, gallocatechins, epigallocatechins and/or afurcatechins, and wherein the proanthocyanidins are proanthocyanidins, prodeldelnidins and/or protogeranium pigments. 4. The product of claim 2 which is a milk drink, soy drink or whey drink having a pH of about 6.8 to about 7.2. 5. The product of claim 2 or 3, which is a milk drink, soy drink or whey drink having a pH of from about 6.9 to about 7.0. 6. The product of claim 4 or 5, which is a milk cocoa beverage containing from about 0.6 milligrams to about 2.0 milligrams of cocoa polyphenols per gram of beverage, wherein said polyphenols include (±)-catechin, (±)-epicatechin and/or proanthocyanidin oligomers. 7. The product of claim 6 which is a skim milk cocoa beverage containing from about 0.6 milligrams to about 2.0 milligrams of cocoa polyphenols per gram of the beverage. 8. The product of claim 6 comprising (i) food acid or food base and (ii) one or more cocoa ingredients having a high cocoa polyphenol content selected from the group consisting of partially defatted cocoa powder, whole Defatted cocoa powder, chocolate liquor, liquid cocoa extract, dry cocoa extract and mixtures thereof. 9. The product of claim 8, wherein said part-skimmed or fully-skimmed cocoa powder contains at least 25 milligrams of cocoa polyphenols per gram of skim cocoa powder and said chocolate liquor contains at least about 10 milligrams of cocoa per gram of skim liquor polyphenols, and the cocoa extract contains at least about 200 milligrams of cocoa polyphenols per gram of dry extract. 10. The product of claim 9, wherein said cocoa powder contains from about 20 mg to 50 mg of cocoa polyphenols, wherein said chocolate liquor contains from about 12 mg to about 25 mg of cocoa polyphenols, and wherein said cocoa extract Contains about 250 mg - about 500 mg of cocoa polyphenols. 11. The product of claim 10, wherein said cocoa powder contains about 25 mg to 35 mg cocoa polyphenols, said chocolate liquor contains about 13 mg to about 17 mg cocoa polyphenols, and said cocoa extract contains about 350 mg - about 500 mg cocoa polyphenols. 12. The product of claim 8, further comprising alkalized cocoa powder, an emulsifier, a sweetener, one or more thickeners, one or more stabilizers, one or more flavorants, and/or vitamin-mineral blend. 13. The product of claim 12, wherein said food acid is citric acid; wherein said food base is sodium hydroxide; wherein said thickener is a mixture of microcrystalline cellulose, carrageenan and carboxymethyl cellulose; wherein said stabilizer is a mixture of citrate and phosphate; and wherein said flavor is chocolate or chocolate containing vanilla, raspberry, blueberry, strawberry, banana, coffee, or mocha. 14. An improved sterilization method for preparing a ready-to-eat or ready-to-drink product containing polyphenols, said method comprising the step of lowering the pH of the product by about 0.2 after mixing and prior to sterilization. 15. The method of claim 14, wherein said product is a milk beverage, a soybean beverage, or a whey beverage; wherein said polyphenols are edible flavan-3-ols, edible proanthocyanidins, or mixtures thereof; and wherein The pH of the beverage is lowered by about 0.4 to about 4.8 to about 6.8. 16. The method of claim 15, wherein said flavan-3-ols are catechin, epicatechin, gallocatechin, epigallocatechin and/or afurcatechin; and wherein said The proanthocyanidins are proanthocyanidins, prodeldelnidins and/or progeranium pigments, and wherein the pH is lowered by about 0.6. 17. The method of claim 15, wherein said product is a low-fat cocoa drink; wherein said flavan-3-ols are (±)-catechin and (±)-epicatechin; and wherein said The proanthocyanidins are proanthocyanidins; and wherein the pH of the beverage is from about 6.8 to about 7.2. 18. A method of preparing a beverage having a pH of 4.8 to about 6.8 and containing one or more proanthocyanidins, said method comprising the steps of: (a) adding one or more proanthocyanidins to a beverage having a pH of about 6.8 to about 7.5; (b) adding a food acid to lower the pH of the beverage by at least about 0.2; (c) heat-treated acidified beverages; and (d) Packaging the heat-treated beverage as a ready-to-drink beverage. 19. The method of claim 18, wherein the pH is lowered by about 0.4 with citric acid. 20. The method of claim 19, wherein the pH is lowered by about 0.6, and wherein the heat treatment is performed at about 145°F to about 280°F for about 1 second to about 30 minutes. 21. The method of claim 20, wherein the heat treatment is performed at about 161°F to about 280°F for about 1 second to about 15 seconds, and wherein the pH of the packaged beverage is from about 6.2 to about 6.8. 22. A method of preparing a cocoa beverage comprising at least about 0.2 milligrams of cocoa polyphenols per gram of beverage, said method comprising the steps of: (a) slurried under high shear with water and a dry cocoa mixture comprising one or more cocoa ingredients selected from the group consisting of (i) cocoa polyphenols containing at least Partially defatted cocoa powder of about 25 mg cocoa polyphenols, (ii) dry cocoa extract having a cocoa polyphenol content of at least 200 mg cocoa polyphenols per gram of extract, (iii) alkalized cocoa powder; (b) blending into milk a dry blend consisting essentially of a sweetener, one or more thickeners, one or more stabilizers, and a vitamin-mineral mixture; (c) adding the cocoa slurry from step (c) to the milk mixture from step (b); (d) adding liquid flavoring; (e) adding edible acid or edible base to the mixture obtained from step (d) to lower the pH by at least 0.2; (f) heat treating the slurry from step (e) at about 161°F to about 280°F for about 1 second to about 15 seconds; (g) optionally cooling and/or homogenizing the slurry obtained from step (f); and (h) packaging the slurry from step (f) or step (g) as a ready-to-drink cocoa beverage. 23. The method of claim 22, wherein prior to said heat treating, said beverage has a water content of at least about 50% by weight and a pH of about 6.8 to about 7.5. 24. The method of claim 22, wherein the beverage has a water content of greater than about 80% by weight, and wherein the pH is from about 6.8 to about 7.5. 25. The method of claim 23, wherein prior to said heat treatment, said pH is from about 6.9 to about 7.0. 26. The method of claim 22, wherein said thickener is gum; wherein said stabilizer is a citrate/phosphate blend; wherein said flavor is chocolate, vanilla, blueberry, raspberry, strawberry , banana, and mocha coffee; wherein the food acid is citric acid; and wherein the food base is sodium hydroxide. 27. The method of claim 22, wherein said cocoa ingredient is prepared from unfermented and/or underfermented cocoa beans. 28. The method of claim 22, wherein the cocoa extract is prepared by solvent extraction of defatted freeze-dried unfermented or underfermented cocoa beans and removal of the solvent. 29. A method of minimizing the loss of (-)-epicatechin and (+)-catechin during heat treatment of a polyphenol-containing product having a water content of at least about 5% by weight, said method comprising A step of lowering the pH of said product by at least 0.2 prior to heat treatment. 30. The method of claim 29, wherein said product is a milk drink, a soy drink, or a whey drink; wherein said polyphenols are cocoa polyphenols; and wherein said pH is lowered by at least 0.4. 31. The method of claim 30, wherein said pH is lowered by at least 0.6; wherein, after said heat treatment, said cocoa polyphenols comprise (+)-epicatechin, (-)-epicatechin, (+)-catechin, (-)-catechin, and proanthocyanidin dimers and trimers.

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