AU2008201286A1 - Compositions containing green tea catechins and one or more polyvalent mineral cations - Google Patents

Description

AUSTRALIA

Patents Act COMPLETE SPECIFICATION

(ORIGINAL)

Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: Name of Applicant: The Procter Gamble Company Actual Inventor(s): Jeffrey John Kester, Ashok Prem Chand Luhadiya, Gerhard Norbert Zehentbauer Address for Service and Correspondence: PHILLIPS ORMONDE FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Invention Title: COMPOSITIONS CONTAINING GREEN TEA CATECHINS AND ONE OR MORE POLYVALENT MINERAL CATIONS Our Ref 825321 POF Code: 103357/44135 The following statement is a full description of this invention, including the best method of performing it known to applicant(s): 00 O COMPOSITIONS CONTAINING GREEN TEA CATECHINS C AND ONE OR MORE POLYVALENT MINERAL CATIONS SThe present application is a divisional application from Australian patent application \number 2004257595, the entire disclosure of which is incorporated herein by reference.

IN FIELD OF INVENTION 00 The present invention relates to compositions containing green tea catechins and one or more polyvalent mineral cations. Such compositions include ready-to-drink (RTD) beverages, as well as powders and concentrates that are combined with an aqueous liquid to form a consumable 00 beverage.

C BACKGROUND Consumption of tea, particularly green tea, has increasingly been associated with therapeutic benefits in the areas of cardiovascular health and cancer prevention. These and other health benefits have been attributed to the unoxidized flavanols in green tea, i.e. the catechin monomers. As a result of these health benefits, there has been a substantial increase in the number of entrants of tea containing beverages into the ready-to-drink beverage market. Currently, many tea containing beverages are sold in the form of bottled, canned or carton-boxed products. Many of the beverages contain black tea extract with fruit juice, fruit aroma or fruit flavors, while others contain only tea flavors. When black tea is used in beverages, many of the published health benefits of the unoxidized flavanols (green tea catechins) are not realized by the consumer because black tea contains oxidized, polymerized flavanols. While certain beverage products contain green tea extract, these beverages generally contain a relatively low level of the beneficial catechins and typically do not contain a significant level of polyvalent mineral cations. The beverages also typically have a brown color or contain natural or artificial colorants to mask the brown color, or the beverages have a turbid appearance. There are not any green tea-containing beverages currently on the market that possess the combination of a relatively high level of catechins, one or more polyvalent mineral cations, and a relatively high degree of clarity low turbidity).

Therefore, there is a need for such green tea-containing beverages.

It is an aspect of this invention to provide compositions that contain significant levels of green tea catechins and one or more polyvalent mineral cations, wherein such compositions, when prepared in a final beverage form, are relatively clear. In one particular embodiment, the final beverage has a low level of brown color such that the appearance of the beverage is not typical of tea, but rather is similar to water.

00 A reference herein to a patent document or other matter which is given as prior art is not to 0 be taken as an admission that that document or matter was, in Australia, known or that the information it contains was part of the common general knowledge as at the priority date of any of Sthe claims.

Throughout the description and claims of the specification, the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other

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00 additives, components, integers or steps.

SSUMMARY OF THE INVENTION 00 0In one aspect, the present invention relates to a process for treating a tea extract, the Sprocess comprising: providing a tea extract in a liquid form, combining the tea extract and a soluble calcium source, permitting the tea extract and soluble calcium source to interact under conditions sufficient to form a precipitate of insoluble calcium components, and separating the insoluble calcium components from the tea extract.

In another aspect, the present invention relates to a fortified ready-to-drink (RTD) beverage composition comprising at least about 100 ppm green tea catechins; and at least about of the U.S. Reference Daily Intake (RDI) of one or more polyvalent mineral cations per 250 mL of the composition; wherein the composition exhibits a turbidity of not more than about 150 Nephelometric Turbidity Units (NTU).

In another aspect, the invention relates to a liquid concentrate or dry beverage composition that exhibits the above characteristics upon dilution with the appropriate amount of aqueous liquid water) to provide a consumable beverage.

00 00 00 00 DETAILED DESCRIPTION OF THE INVENTION A. Definitions As used herein, the term "botanical flavor" or "botanical extract" refers to flavors derived from parts of the plant other than the fruit. As such, botanical flavors can include those flavors derived from nuts, bark, roots and leaves. Also included within this term are synthetically prepared flavors made to simulate botanical flavors derived from natural sources. Examples of botanical flavors include hibiscus, marigold, chrysanthemum and the like. Botanical flavors can also be synthetically prepared.

As used herein, the term "caloric content" refers to the total energy in kilocalories (kcal) per serving of consumable beverage (250 mL) derived from the carbohydrate, protein, and lipid components of the beverage. Caloric content is calculated using the general factors of 4, 4, and 9 kcal per gram for the total level of carbohydrate, protein, and lipid, respectively (United States Code of Federal Regulations, Title 21, Section 10 1.9, Nutrition Labeling of Food).

As used herein, the term "comprising" means various components can be conjointly employed in the beverages of this invention. Accordingly, the terms "consisting essentially of' and "consisting of' are embodied in the term comprising.

As used herein, the terms "concentrated liquid composition" and "concentrate" mean a liquid composition of the present invention that is intended to be diluted with an aqueous liquid, such as water, prior to consumption as a beverage. The skilled artisan will recognize that the concentration of such compositions will be dictated by the components contained therein and their relative stability in an aqueous state. The concentrates of the present invention are reconstituted with an aqueous liquid, such as water, to provide a consumable beverage as described herein.

As used herein, the term "consumable beverage" and "final beverage" are used interchangeably and refer to a beverage that is prepared diluted) in accordance with relevant WO 2005/006871 PCT/US2004/020872

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00 3 c package labeling. Thus, as discussed below, a ready-to-drink beverage is one that is sold such Sthat it is consumed without additional preparation by the consumer. In contrast, dry beverage compositions and concentrates are intended to be diluted to provide a consumable beverage.

As used herein, the term "dry beverage composition" means a flowable particulate IsO compositions that is substantially free of moisture a powder). Typically, such compositions 00 01 will contain not more than about 6% by weight total moisture. The dry beverage compositions of the present invention are reconstituted with an aqueous liquid, such as water, to provide a 00 consumable beverage as described herein.

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SAs used herein, the term "juice" means whole, concentrated or diluted juice from fruits C,1 and vegetables and other produce which are squeezed or crushed to supply a beverage. Juice also refers to citrus and non-citrus juices including vegetable juices.

As used herein the term "fruit flavors" refers to those flavors derived from the edible reproductive part of the seed plant, especially one having a sweet pulp associated with the seed, for example, apples, oranges, lemon, limes, etc. Also included within the term fruit flavor are synthetically prepared flavors made to simulate fruit flavors derived from natural sources.

As used herein, the term "green tea catechins" refers to the total content in parts per million (ppm) (or mg/L or ug/mL) of the following five catechins derived from green tea: epicatechin epigallocatechin (EGC), epigallocatechin gallate (EGCG), gallocatechin gallate (GCG), and epicatechin gallate (ECG).

As used herein, the term "nutritionally-supplemental amount" is meant that the mineral (and any optional vitamin) sources used in the practice of this invention provide a nourishing amount of minerals and vitamins. This supplemental amount will provide from about 5% to about 150% of the U.S. Reference Daily Intake (RDI). The RDI for vitamins and minerals is as defined in the United States of America (United States Code of Federal Regulations, Title 21, Section 101.9, Nutrition Labeling of Food).

As used herein, the term "polyvalent mineral cation" refers to one or more minerals that typically exist in solution in the form of positively charged cations with a net charge of +2 or greater. Non-limiting examples of polyvalent mineral cations include calcium (Ca 2 magnesium (Mg 2 manganese (Mn 2 zinc (Zn 2 and iron (Fe 2 Fe 3 As used herein, "ready-to-drink" (or "RTD") and "ready-to-serve" are used interchangeably and refer to compositions of the present invention that are at their recommended drink strength and, therefore, are ready for consumption without the need for reconstitution or dilution with aqueous liquids.

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As used herein "single strength" refers to recommended drink strength, i.e. the recommended concentration of the given beverage composition.

As used herein the term "shelf stable" refers to packaged beverages that do not require refrigeration while stored at ambient temperatures for prolonged periods more than 10 days).

As used herein, the term "solids" refers to the solid materials extracted from the tea 00 solutions which are soluble in water. The solids include caffeine, flavanols, amino acids (especially theanine), carbohydrates, edible acids, buffering salts, proteins and related materials.

00 As used herein, the term "total insoluble calcium component" refers to the aggregate of Sthe moieties of a sample that bind to calcium under the conditions described in the Analytical Ci Methods section below (Measurement of the Total Insoluble Calcium Components) and thereby become insoluble in the sample.

As used herein, the term "vitamin C" refers to L-ascorbic acid. The term "erythorbic acid" refers to its isomer, D-isoascorbic acid.

As used herein, the term "water" includes any of tap, filtered, purified, deionized, demineralized, distilled, spring, mineral, artesian, softened, sparkling, or carbonated water.

All percentages are by weight unless otherwise indicated.

B. Compositions All concentration limits of components provided herein are based on single strength beverages, unless otherwise stated.

As mentioned, the invention relates to RTD beverages, as well as concentrated liquids and dry beverage compositions that exhibit the above characteristics upon dilution with the appropriate amount of aqueous liquid to provide a consumable beverage. Thus, in the case of dry or concentrate compositions, it will be recognized that the levels described presume that the dry or concentrate composition is diluted to provide a consumable beverage. That is, where catechin, polyvalent cation, turbidity, etc. levels are stated with respect to a dry or concentrate composition, those levels refer to the composition after it is appropriately diluted to provide a consumable beverage.

The compositions of the present invention comprise a significant level of green tea catechins, which are known to be associated with various health benefits. Some of these health benefits include cancer prevention, improved cardiovascular health, and reduced blood pressure.

The compositions of the present invention comprise at least about 100 ppm of green tea catechins.

(Again, for compositions that are intended to be diluted prior to consumption, the level of catechins stated herein assumes dilution as appropriate for such consumable beverage.) In one aspect, the beverage composition will comprise at least about 150 ppm green tea catechins. In WO 2005/006871 PCT/US2004/020872

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00 Sanother aspect, the beverage composition will comprise at least about 200 ppm green tea Scatechins. Preferably, the beverage composition will comprise at least about 300 ppm green tea catechins, and most preferably at least about 400 ppm green tea catechins. Typically, the beverage composition will comprise from about 100 to about 3,000 ppm green tea catechins.

O The beverage compositions of the present invention have a relatively high level of clarity 00 Ci low turbidity). In this regard, the compositions (in final, consumable form) will exhibit a turbidity of not more than about 150 NTU. In another aspect, the compositions will typically c exhibit a turbidity of not more than about 125 NTU, more typically not more than about 100 SNTU. Preferred beverage compositions of the present invention will exhibit a turbidity of not C'1 more than about 50 NTU, even more preferably not more than about 30 NTU. Typically, the beverage compositions will exhibit a turbidity of from about 1 to about 150 NTU. Turbidity of the consumable beverages is measured with a laboratory turbidimeter as described in the Analytical Methods section.

The finished beverage compositions will comprise, on a 250 mL serving basis, at least about 10% of the RDI of one or more polyvalent mineral cations. While the polyvalent mineral cation(s) may be selected from a wide range of materials, the following provides a non-exhaustive list of preferred cations. Preferred polyvalent mineral cations may be selected from calcium, magnesium, manganese, zinc, and/or iron (di- or trivalent), or mixtures thereof. The RDI values for these minerals are as follows (United States Code of Federal Regulations, Title 21, Section 101.9, Nutrition Labeling of Food): calcium, 1000 mg; magnesium, 400 mg; manganese, 2 mg; zinc, 15 mg; and iron, 18 mg. Suitable sources of these minerals include, but are not limited to, the following salts: citrate, sulfate, gluconate, lactate, bis-glycinate, amino acid chelates, carbonate, oxide, hydroxide, chloride, phosphate, pyrophosphate, acetate, fumarate, and malate.

Preferred sources of calcium are calcium citrate malate, calcium gluconate, calcium lactate, and calcium lactate gluconate. Preferred sources of magnesium are magnesium sulfate and magnesium gluconate. A preferred source of zinc is zinc gluconate. A preferred source of iron is the amino acid chelate iron bis-glycinate, available as Ferrochel® from Albion Laboratories, Inc., Clearfield, Utah. Another preferred source of iron is ferrous gluconate. Optionally, other minerals can be added to the beverage compositions of the present invention, including phosphorus, iodine, selenium, copper, fluoride, chromium, molybdenum, sodium, potassium, and chloride. In one aspect, the compositions will comprise, on a 250 mL serving basis, at least about of the RDI of one or more polyvalent mineral cations. In yet another aspect, the compositions will comprise, on a 250 mL serving basis, at least about 20% of the RDI of one or more polyvalent mineral cations. Typically, the compositions will comprise, on a 250 mL serving WO 2005/006871 PCT/US2004/020872 00 6 C basis, from about 10% to about 150% of the RDI of one or more polyvalent mineral cations. Of Scourse, the dry and concentrate compositions of the present invention will have mineral levels such that they provide the above described ranges upon appropriate dilution to provide a consumable beverage.

IDThe compositions will preferably have, when in final form for consumption, a pH of from 00 Sabout 2 to about 6.5. In one aspect, the consumable beverage will have a pH of from about 2.5 to O about 5, more preferably from about 3 to about 4.5. A pH within the most preferred range of from N about 3 to about 4.5 is particularly desirable for minimizing dark color in a consumable beverage that does not have added coloring agents. For example, applicants surprisingly discovered that, Seven when iron is not part of the formulation, green tea and mineral fortified water beverages are characterized by significantly less brown color when the pH is within the most preferred range.

When iron is one of the fortifying polyvalent mineral cations in the beverage formulation, low pH is especially important for minimizing the interaction between the iron cation and green tea polyphenol components, which yields significant darkening of the beverage at higher pH.

The compositions of the present invention will preferably exhibit low brown color, which is important to the ultimate consumer. Brown color is characterized by measuring absorbance of the consumable beverage at 430 nmn as described in the Analytical Methods section. (Again, it will be recognized that the absorbance at 430 nm is measured using the consumable beverage.

Thus, for compositions that are intended to be diluted prior to consumption, the absorbance characteristics stated herein assume dilution as appropriate for such consumable beverage.) Absorbance at 430 nm reflects the degree of brownish color in the beverage that can be caused by the presence of theaflavins, thearubigins, proteins, chlorophylls, and their degradation products.

In one aspect, the beverage compositions will preferably have an absorbance at 430 nm of not more than about 0.15. In another aspect, the beverage compositions will have an absorbance at 430 nmn of not more than about 0.12. In yet another aspect, the beverage compositions will have an absorbance at 430 nm of not more than about 0.10. Preferably, the beverage compositions will have an absorbance at 430 nm of not more than about 0.08, most preferably not more than about 0.05. Typically, the beverage compositions will have an absorbance at 430 nm of from about 0.01 to about 0.15.

It will be recognized that in addition to preferably being low in brown color, by virtue of low absorbance at 430 nm, the beverages of the present invention will also preferably be low in color in general. As absorbance at 430 nm of the finished beverage decreases, appearance of the beverage becomes more similar to water and eventually the appearance becomes almost WO 2005/006871 PCT/US2004/020872 00 7 Sindistinguishable from water. This is particularly true when the beverage is packaged in a green colored glass or plastic bottle, as is often the case for premium bottled waters Perrier®).

Applicants have discovered that undesired solubility issues associated with inclusion of calcium (often a preferred nutrient in fortified beverages) in the present beverage compositions I\ can be addressed by limiting the level of total insoluble calcium components, including oxalic 00 00 acid, prior to calcium addition. As such, compositions of the present invention that include calcium will preferably contain not more than about 35 ppm oxalic acid or salts thereof. More 00 typically, the compositions will contain not more than about 20 ppm, still more typically not more Sthan about 7 ppm, oxalic acid or salts thereof. In addition, when calcium is present in the C( beverage composition, the composition will preferably contain not more than about 1000 ppm of total insoluble calcium components including oxalic acid. More typically, the compositions will contain not more than about 450 ppm, still more typically not more than about 250 ppm, total insoluble calcium components including oxalic acid. A preferred process for removing total insoluble calcium components, including oxalic acid, to avoid solubility issues is described below.

In a preferred process, tea extract is treated with a soluble calcium source to remove insoluble calcium components without affecting the level of catechins. The treatment is typically carried out at ambient or higher temperatures, preferably at temperatures greater than about 130°F More typically, the reaction temperature is greater than about 145°F still more typically the reaction temperature is greater than about 160 0 F The reaction time is typically about 2 hours or less. More typically the reaction time is about 45 minutes or less; still more typically the reaction time is about 15 minutes or less. The preferred calcium salts to promote precipitation of the calcium insoluble components are calcium sulfate, calcium chloride, calcium acetate and calcium gluconate. Other sources of soluble calcium can also be used. The preferred pH during the treatment is less than or equal to about 7. The concentration of soluble calcium added to the tea extract to promote precipitation of the calcium insoluble components may vary from about 400 mg to about 10,000 mg per 100 gm of tea extract on a dry weight basis.

For conventional green tea extracts, e.g. green tea powdered extract #285060 commercially available from Plantextrakt, Inc. (Parsippany, NJ), the amount of calcium required tends to be toward the upper limit of this range of addition. For nano-filtered tea extract, e.g. Green Power SynerTea T M brand of green tea extract (commercially available from Amax NutraSource, Inc., Eugene, OR), the amount of calcium required tends to be toward the lower limit of this range of addition. This is because the nanofiltration process used during manufacture of the SynerTea T M brand of green tea extract removes a portion of the higher molecular weight calcium insoluble WO 2005/006871 PCT/US2004/020872 00 8 Scomponents, such as pectin. After treatment to precipitate the calcium insoluble components, the Stea extract is filtered to remove the insoluble/suspended material. The preferred level of insoluble calcium components in the treated green tea extract, on a dry weight basis, is not more than about 1% w/w. More typically, the level of insoluble calcium components is not more than about 0.6% IND w/w; still more typically the level is not more than about 0.1% w/w.

00 In a related aspect, applicants have surprisingly discovered that the solubility issues associated with calcium addition are not encountered with other polyvalent mineral cations. As 00 such, it surprisingly does not appear that additional processing of the green tea to reduce oxalic acid/pectin levels) is necessary when soluble polyvalent mineral cations other than calcium CK1 are included in the present compositions.

The beverage compositions of the present invention will preferably have a relatively low caloric content. In one preferred aspect of the invention, the compositions will have a caloric content, on a 250 mL serving basis, of not more than about 60 kilocalories (kcal). In another aspect, the compositions will typically have a caloric content, on a 250 mL serving basis, of not more than about 30 kcal, preferably not more than about 15 kcal. In a similar regard, the beverage compositions will preferably have a relatively low carbohydrate level. As such, in one preferred aspect, the compositions will have a carbohydrate level, on a 250 mL serving basis, of not more than about 15 g, preferably not more than about 8 g, still more preferably not more than about 4 g.

The beverage compositions of the present invention, when in the form of a liquid concentrate or a RTD beverage, comprise water. Ready-to-drink beverages typically contain from about 80% to about 99.8% water. Preferably the water is soft, demineralized, or deionized.

Concentrates of the present invention typically contain from about 20% to about 79%, more typically from about 30% to about 70% water. If desired, the water used in the beverage formulation may be carbonated; e.g. sparkling or carbonated water. More typically, a carbonated beverage is produced by solubilizing carbon dioxide gas into the final blended beverage. Usually a beverage will be considered carbonated if it comprises more than 30%, preferably more than 100%, by volume of solubilized carbon dioxide. Carbonated beverages typically comprise from 100% to 450%, preferably from 200% to 350%, carbon dioxide by volume of the beverage.

The beverage compositions of the present invention comprise green tea extract. The extract can be obtained from either a single tea plant or a mixture of plants and it can be in the form of a liquid extract or a dry powdered extract. It is preferred that the green tea extract be light in color and enriched in unoxidized flavanols, catechin monomers. Preferably, the green tea extract comprises at least about 10% green tea catechins on a dry solids basis, more preferably at least about 20% catechins. Non-limiting examples of suitable green tea extracts include Green WO 20105/006871 PCT/US2004/020872 00 9

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SPower SynerTea T M brand of green tea extract (commercially available from Amax NutraSource, SInc., Eugene, OR), green tea powdered extract #285060 commercially available from Plantextrakt, Inc. (Parsippany, NJ), and green tea extracts produced by the processes described in U.S. Patent No. 5,879,733 issued to Ekanayake, et al. (1999) and U.S. Patent No. 5,427,806 IN) issued to Ekanayake, et al. (1995). Preferred green tea extracts to use when calcium is not 00 Sincluded in the beverage formulation are the Green Power SynerTeaTM brand of green tea extract and/or the extract prepared according to the process described in U.S. Patent No. 5,879,733. A 00 preferred green tea extract when calcium is included in the beverage compositions of the present Sinvention is the extract prepared according to the process described above, in which the extract CNl has been treated to remove insoluble calcium components, such as oxalic acid and pectin. The level of green tea extract in the beverage compositions of the present invention may vary, as long as the beverage comprises at least about 100 ppm of green tea catechins. Typically, the consumable beverages of the present invention will comprise from about 0.03% to about 1% green tea extract solids, more typically from about 0.08% to about In one aspect of the present invention, the beverage composition preferably comprises food-grade acids to reduce the pH of the beverage. The inventors surprisingly discovered that addition of acid to reduce the pH of the consumable beverage results in a lower absorbance at 430 nm, i.e. the beverage is less brown in color. Non-limiting examples of food-grade acids that can be used to reduce the pH of the beverage include citric, malic, phosphoric, tartaric, ascorbic, and erythorbic acids. Another benefit associated with the addition of food-grade acids to the present beverage composition is improved color stability of the beverage during storage. It is known that the brown color of tea beverages results from oxidation of the catechin monomers to yield highmolecular weight polymers theaflavins and thearubigins). Oxidation of the catechins can be controlled and, hence, browning of the green tea beverage delayed or prevented by the addition of acids such as citric, ascorbic, and erythorbic acids. While not wishing to be bound by theory, it is believed that citric acid helps prevent oxidation of the green tea catechins by lowering the beverage pH and by complexing metal ions such as iron and copper, which are known to be oxidation catalysts. It is believed that ascorbic and erythorbic acids help prevent oxidation of the catechins because they are effective reducing agents. The preferred food-grade acids for use in the beverage compositions of the present invention are citric, malic, ascorbic, and/or erythorbic acids.

Other optional ingredients in the beverage compositions of the present invention include carbohydrate sweeteners, zero or low calorie high-intensity or artificial sweeteners, soluble fibers and polysaccharides, proteins and/or peptides, amino acids, emulsifiers, vitamins, salts, natural WO 2005/006871 PCT/US2004/020872 00 t and/or artificial flavoring agents, flavor enhancers, caffeine, preservatives, and natural and/or Sartificial coloring agents or other ingredients.

_The beverage compositions of the present invention can optionally contain a natural and/or artificial flavoring agent. The particular amount of flavoring agent effective for imparting IDflavor characteristics to the beverage depends upon the flavor(s) selected, the flavor impression 00 desired and the form of the flavor. The flavoring agent can comprise a fruit juice or vegetable juice, a fruit or vegetable flavor or mixtures thereof. The juice can be provided as juice made 00 from, for example, apple, cranberry, pear, peach, plum, apricot, nectarine, grape, cherry, currant, raspberry, gooseberry, blackberry, blueberry, strawberry, lemon, lime, mandarin, orange, grapefruit, potato, tomato, lettuce, celery, spinach, cabbage, watercress, dandelion, rhubarb, carrot, beet, cucumber, pineapple, coconut, pomegranate, kiwi, mango, papaya, banana, watermelon, tangerine and cantaloupe. Preferred juices are apple, pear, lemon, lime, mandarin, grapefruit, cranberry, orange, strawberry, tangerine, grape, kiwi, pineapple, passion fruit, mango, guava, raspberry and cherry. Citrus juices, such as grapefruit, orange, lemon, lime, mandarin and juices of mango, passion fruit and guava, or mixtures thereof, are most preferred.

The fruit or vegetable juice(s) are present in an amount of from 0% to about 100%, preferably from 0% to about 35%, and more preferably from 0% to about 10% of the beverage.

This concentration is based on the single strength of the beverage.

The flavoring agent according to the present invention can also comprise flavors selected from fruit flavors, botanical flavors, vegetable flavors and mixtures thereof Particularly preferred fruit flavors are the citrus flavors including orange flavors, lemon flavors, lime flavors and grapefruit flavors. Besides citrus flavors, a variety of other fruit flavors can be used such as apple flavors, grape flavors, cherry flavors, pineapple flavors and the like. These fruit flavors can be derived from natural sources such as fruit juices and flavor oils, or else be synthetically prepared.

Particularly preferred botanical flavors are hibiscus, marigold and chrysanthemum.

The flavor agent can also comprise a blend of various flavors, lemon and lime flavors, citrus flavors and selected spices (the typical cola soft drink flavor) etc. If desired the flavor can be formed into emulsion droplets and then dispersed in the beverage drink. Because these droplets usually have a specific gravity less than that of water and would therefore form a separate phase, weighting agents (which can also act as clouding agents) can be used to keep the emulsion droplets dispersed in the beverage. Examples of such weighting agents are sucrose acetate isobutyrate, brominated vegetable oils (BVO) and resin esters, in particular the ester gums.

See L.F. Green, Developments in Soft Drinks Technology,. Vol. 1 (Applied Science Publishers Ltd. 1978) pp. 87-93 for a further description of the use of weighting and clouding agents in liquid beverages.

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0 SFlavoring agents which are not substantially juice, comprise no more than about 3% of Sthe beverage, preferably such flavor component comprises at least 0.001% by weight of the beverage and typically from about 0.01% to about 3% by weight of the beverage.

The beverage compositions of the present invention can optionally contain preservatives such as benzoic acid, sorbic acid, and salts thereof; salts such as sodium chloride, potassium 00 00 chloride, sodium phosphate, potassium phosphate, sodium citrate, and potassium citrate; polysaccharides and/or soluble fibers such as guar gum, xanthan gum, alginates, starches, pectin, c maltodextrins, gum arabic, arabinogalactan, inulin, fructooligosaccharide, and polydextrose; Sproteins such as milk proteins (caseins and/or whey proteins) and soy protein; emulsifiers such as mono- and di-glycerides and lecithin; and vitamins such as vitamins C, A, D, E, K, and the Bcomplex vitamins BI (thiamine), B2 (riboflavin), B3 (niacin), B6 (pyridoxine), B12 (cyanocobalamin), and folic acid.

The beverage compositions of the present invention are produced by combining the appropriate ingredients using standard beverage manufacturing and processing procedures. For example, see The Chemistry and Technology of Soft Drinks and Fruit Juices Ashurt, Editor, CRC Press, 1998). In making a single strength beverage, a liquid concentrate or syrup is usually formed first. This liquid concentrate typically contains flavoring agents and/or fruit juice concentrates, acids, salts, high-intensity sweeteners if used, and suitable preservatives if needed.

The liquid concentrate usually contains from about 30% to about 70% by weight water. The ingredients are added to the water and mixed in conventional equipment. If desired, the green tea extract can also be incorporated into the liquid concentrate or, alternatively, the green tea extract may be added to the final blended beverage just prior to packaging. After the concentrate is formed, additional carbohydrate sweetener may be added and water is added to make a finished consumable beverage ready for packaging. The product may be cold-filled or hot packed into a suitable container.

C. Analytical Methods 1. Measurement of the Green Tea Catechin Level: The total level of green tea catechins in consumable beverages or aqueous tea extracts is measured using the following method.

a) Reference: A. Finger et al., "Chromatography of tea constituents", J. Chromatogr., (1992), 624: 293-315.

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c This method is used to measure the level of green tea catechins in consumable beverages.

SFive green tea catechins (EC, EGC, EGCG, GCG, and ECG) are measured in the beverage using linear response plots prepared with individual pure standards. This method is also used to measure the level of green tea catechins in aqueous tea extracts \after dissolving or diluting the tea extract in water to achieve approximately 0.15% tea 00 00 solids.

c) Principle: (1 The consumable beverage is usually diluted 1:10 or 1:100 with diluent (see below) to 00 obtain signals lying within the range of the calibration curve. The diluted sample is C< filtered through a 0.2 gm filter to remove particulates. The sample is then injected onto a C-18 reversed phase High Performance Liquid Chromatography (HPLC) column. The catechins are eluted using a gradient mobile phase composed of a 0.01 M phosphate buffer at pH 3.1 and acetonitrile. Detection is performed using a UV absorbance detector set to 280 nm.

d) Apparatus: Class A volumetric flasks (1000 mL, 10 mL, 5 mL) Sartorius four place balance Supelco vacuum filtration unit HP auto sampler vials Whatman Anotop 25 (0.2 pm) inorganic membrane filter mL disposable syringe Instrument Set-up Agilent-1090 HPLC Agilent 1090 Diode Array Detector set to 280 nm Agilent Chemstation for LC, Rev. A.04.02 YMC basic column 250 mm x 2 mm 5 im, serial 112099G e) Reagents: Acetonitrile (HPLC grade, J. T. BAKER, cat# 9011-03, lot N42B26) Milli-Q water with resistance greater than or equal to 18 MQ-cm EDTA (Fisher Scientific, cat#. S311, lot 860954) Phosphoric acid T. BAKER, cat# 0260-01, lot 25806) Potassium dihydrogen phosphate T. BAKER, cat# 4008-01, lot 30K0178)) Sodium bisulfite (Sigma, cat#. S9000, lot 30K0178) WO 2005/006871 PCT/US2004/020872 00 13 SCaffeine (CF, Sigma, cat#. C0750, lot 127F-0395) SEpigallocatechin (EGC, Sigma, cat#. E3768, lot 61K1428) Epicatechin (EC, Sigma, cat#. E1753) Epigallocatechingallate (EGCG, Sigma, cat#. E4143, lot 61K1773) Epicatechingallate (ECG, Sigma, cat#. E3893, lot 31K1236) 00 00 Gallocatechin gallate (GCG, Sigma, cat#. G6782, lot 110K1413) f) Solutions: 0 0.01 M (1.3i g/L) potassium dihydrogen phosphate buffer: 00 O Weigh 1.36 g potassium dihydrogen phosphate into a 1 liter volumetric flask. Add CN approximately 600 mL HPLC grade water and stir until solid is dissolved. Titrate with phosphoric acid to a pH of 3.1. Bring to volume with HPLC grade water. Degas using vacuum filtration system.

Diluent: Use to dilute the sample and ensure stability of the catechins under ambient conditions.

Add 0.24 g EDTA and 1.24 g sodium bisulfite (NaHSO 3 to 910 mL of 0.01M potassium dihydrogen phosphate buffer. Add 90 mL of acetonitrile.

Preparation of beverage sample: Typically, pipet 0.5 mL of a finished beverage into a 5 mL volumetric flask (1:10 dilution) or 0.1 mL of a finished beverage into a 10 mL volumetric flask (1:100 dilution) and bring to volume with diluent. Filter the resulting solution through a 0.2 pm Whatman filter. Transfer the filtered sample to the auto sampler vial.

Preparation of standard solutions: Each pure standard (caffeine and the five catechins) is accurately weighed and placed into a 5 mL or 10 mL volumetric flask as shown below. Add diluent to dissolve the standard and fill the flask to volume. These are standard stock solutions (~1000 ppm). Standard working solutions are then prepared by diluting the standard stock solutions with diluent.

The actual weights and concentrations of the standard stock solutions and working solutions are shown in the following Table.

EGC CF EC EGCG GCG ECG Volume (mL) 5 10 10 5 5 Actual weights 0.004 0.0105 0.094 0.0049 0.0045 0.0044 3 0 Stock Solution 860 1050 940 980 900 880 WO 2005/006871 PCT/US2004/020872 (pg/mL) Working 172 420 188 196 180 176 Solution (Ag/mL) Using the standard working solutions, response factor mixtures are prepared as shown below: iL of Standard Working Solution

NO

00 00

(N

00 Mixture# EGC Caffeine EC EGCG ECG GCG pL Diluent Total PL 6 600 400 250 100 250 250 3150 5000 7 500 500 20 300 150 20 3510 5000 8 400 25 100 1200 200 100 2975 5000 9 150 300 200 900 20 200 3230 5000 50 100 300 650 300 300 3150 5000 The actual concentrations of each standard (caffeine and the 5 catechins) factor mixtures are as follows: in the response Concentration of Standards (p.g/mL) Mixture EGC Caffeine EC EGCG ECG GCG 6 20.64 33.6 9.4 3.92 8.8 7 17.2 42 0.75 11.76 5.28 0.72 8 13.76 2.1 3.76 47.04 7.04 3.6 9 5.16 25.2 7.52 35.28 0.704 7.2 1.72 8.4 11.28 31.36 10.56 10.8 g) Chromatography Conditions: Sample size 25 tL Detector -280 nm AUFS 0.100 Mobile Phase: A water B 0.01M potassium dihydrogen phosphate buffer, pH 3.1 C Acetonitrile Gradient: 0 min. 1%A 90% B 9%C min. 2% A 80%B 18% C 18 min. 3% A 70%B 27% B WO 2005/006871 00 Os PCT/US2004/020872 min. 1% A 90% B 9% C min. stop Flow rate: 0.4 mL/min 00 h) Calculation of Catechin Level in the Consumable Beverage: 00 Peak areas corresponding to each catechin standard on the HPLC chromatogram of the 0response factor mixtures are used to prepare a linear response plot for each catechin. The 00 correlation coefficients (r-square) for each response plot are typically equal to or greater than 0.99. The beverage sample chromatogram is analyzed for the peak area of each catechin. The level of catechins in the beverage sample is determined from the linear response plots. The level of catechins in the consumable beverage is then calculated by multiplying by the dilution factor used (10x or 100x). The total level of green tea catechins in ppm (gg/mL or mg/L) in the finished beverage is calculated by adding the levels of the five individual catechins measured (EGC EC EGCG ECG GCG).

2. Measurement of the Turbidity of Consumable Beverages: A Hach Model 2100AN Turbidimeter (Hach Company, Loveland, CO) is used to measure the turbidity of consumable beverages. This instrument measures the turbidity of liquids in Nephelometric Turbidity Units (NTU). The turbidimeter is first calibrated with a set of Gelex® Secondary Turbidity Standards. Measurements are made using the following instrument parameters: ratio on; signal averaging on; automatic range selection; US EPA filter installed. The beverage is equilibrated to room temperature and then poured into a turbidimeter sample cell (sample cell outer diameter 25 mm). The sample cell is inserted into the instrument and the turbidity reading (NTU) is recorded after waiting 30-60 seconds for the reading to stabilize. The measured NTU value is indicative of the level of turbidity of the consumable beverage. Higher NTU values indicate greater turbidity.

3. Measurement of Absorbance at 430 nm of Consumable Beverages: A UV-Visible spectrophotometer (Unicam UV1 UV-Visible Spectrometer; Unicam Limited, Cambridge, United Kingdom) is used to measure absorbance at 430 nm of consumable beverages. The beverage is equilibrated to room temperature and then poured into a sample cuvette (1 cm path length), which is placed in the sample holder of the spectrophotometer. A blank cuvette filled with deionized water is placed in the blank holder of the spectrophotometer. Absorbance of the beverage sample at a wavelength of 430 nm is measured and recorded The cuvette in the sample holder is then replaced with another WO 2005/006871 PCT/US2004/020872 00 16 0 ct blank cuvette filled with deionized water and absorbance of the blank measured at 430 nm and Srecorded The absorbance of the beverage sample is then adjusted by subtracting the measured absorbance of the deionized water blank: Absorbance at 430 nm (B) IN This procedure corrects for any measured absorbance contributed from the water. In theory, 00 absorbance of the deionized water blank will approach zero. The absorbance at 430 nm is indicative of the level of brown color in the beverage.

00 4. Measurement of pH: SThe pH of consumable beverages and aqueous tea extracts is measured with a Coming Model 7. 440 pH Meter. Prior to making pH measurements, the meter is calibrated using pH 4.00 and 7.00 standard buffer solutions (VWR Scientific, West Chester, PA).

Measurement of the Level of Polyvalent Mineral Cations In Consumable Beverages: Levels of the minerals calcium, magnesium, manganese, zinc, and iron in consumable beverages are measured by Inductively Coupled Plasma Emission Spectrometry according to the methods described in the following references: a) "Inductively Coupled Plasma-Atomic Emission Spectrometry Analysis of Biological Materials and Soils for Major, Trace, and Ultra-Trace Elements", Applied Spectroscopy, volume 32, pg. 1-29, 1978.

b) AOAC Official Method 985.01, "Metals and Other Elements in Plants and Pet Foods", (3.2.06, Chp. 3, pg. 4).

c) AOAC Official Method 984.27, "Calcium, Copper, Iron, Magnesium, Manganese, Phosphorus, Potassium, Sodium, and Zinc in Infant Formula", (50.1.15, Chp. 50, pg.

15-16).

All AOAC (Association of Official Analytical Chemists) published methods can be found in the following reference: AOAC International, Official Methods of Analysis. P. Cunniff 16 th edition, 5 t Revision, 1999, Gaithersburg, MD The level of each mineral is expressed as mg per 250 mL of beverage. The %RDI of each mineral is calculated by dividing the mineral level by the respective RDI value: %RDI [(mg/250 mL beverage) (RDI)] x 100 The RDI values for the minerals are as follows (United States Code of Federal Regulations, Title 21, Section 101.9, Nutrition Labeling of Food): calcium 1,000 mg; magnesium 400 mg; manganese 2 mg; zinc 15 mg; iron 18 mg.

WO 20()5/00(16871 PCT/US2004/020872 00 17 t 6. Measurement of the Caloric Content and Carbohydrate Content of Consumable SBeverages: SCaloric content in kilocalories (kcal) per 250 mL of beverage is determined from the carbohydrate, protein, and lipid contents of the beverage. Caloric content is calculated using the general factors of 4, 4, and 9 kcal/g for the content of carbohydrate, protein, and lipid, 00 0C respectively, per 250 mL serving of beverage (United States Code of Federal Regulations, Title 21, Section 101.9, Nutrition Labeling of Food): OO Caloric Content (kcal/250 mL beverage) carbohydrate/250 mL) x 4] protein/250 mL) x 4] lipid/250 mL) x 9] Protein content (g/250 mL beverage) is measured according to the following method (a conversion factor of 6.25 is used to convert nitrogen to protein): AOAC Official Method 968.06, "Protein (Crude) in Animal Feed" (modified), Official Methods of Analysis, Patricia Cunniff 16 th edition, Vol. 1, AOAC International, Gaithersburg, MD (1995).

Lipid content (g/250 mL beverage) is measured by acid hydrolysis according to the following method: AOAC Official Method 954.02, Official Methods of Analysis, 17" edition, Vol. 1, AOAC International, Gaithersburg, MD (2000).

Ash content (g/250 mL beverage) is measured according to the following method: AOAC Official Method 923.03, "Ash of Flour" (modified), Official Methods of Analysis, Patricia Cunniff 16'h edition, Vol. 2, AOAC International, Gaithersburg, MD (1995).

Moisture content (g/250 mL beverage) is measured according to the following method: AOAC Official Method 934.06, "Moisture in Dried Fruits" (modified), Official Methods of Analysis, Patricia Cunniff 16" edition, Vol. 2, AOAC International, Gaithersburg, MD (1995).

Carbohydrate content is calculated by difference using the following equation: Carbohydrate Content (g/250 mL beverage) 100 (Protein Content) (Lipid Content) (Ash Content) (Moisture Content) 7. Measurement of the Moisture Content of Dry Beverage Compositions: Moisture content of dry beverage compositions is measured according to the following method: WO 2005/006871 PCT/US2004/020872 18 AOAC Official Method 979.12, "Moisture (Loss on Drying) in Roasted Coffee", Official SMethods of Analysis, Patricia Cunniff 16 t h edition, 5 th revision, AOAC International, SGaithersburg, MD (1999).

8. Measurement of the Oxalic Acid Level: This method is used to measure the level of oxalic acid in consumable beverages and aqueous 00 CI tea extracts.

O a) Reference: 00 Official Methods of Analysis of AOAC INTERNATIONAL (2000) 17 Ed. AOAC INTERNATIONAL, Gaithersburg, MD, USA, Official Method 986.13. (modified) I b) Scope: This method is used to measure the level of oxalic acid, fumaric acid, tartaric acid, citric acid, malic acid, lactic acid, succinic acid, and acetic acid in food products.

c) Principle: The sample is filtered to remove interferences. Organic acids are then separated using HPLC utilizing UV detection at 214 nm.

d) Apparatus: Class A volumetric flasks (1000 mL, 10 mL, 5 mL) Sartorius four place balance Liquid Chromatograph System equipped with Model 7725i inhector, Model 2487 variable wavelength detector operable at 214 nm, 0.1 AUFS (Waters Associates, Inc.) and computing integrator (Hewlett-Packard Integrator 3390 [replaced by No. 3396B], or equivalent).

Analytcal Columns Supelcosil LC-18, or equivalent, 5 pm particle size, 25 cmX 4.6 mm, in tandem with and followed by Radial-Pak C18 cartridge (Water Associate, Inc.), 5 pm particle size, 10 cm long, used with Radial Compression Module. Radial-Pak C18 cartridge can be substituted by any standard 25 or 30 cm stainless steel reverse phase C18 column with 10 pm particle size. Connect Bio-Rad reverse phase micro-guard column (ODS-10) ahead of column 1. Mobile phase: phosphate buffer at 0.8 mL/min; sensitivity 0.1 AUFS.

e) Disposable cartridges: Sep-Pak C18 (Water Associates, Inc.) 0.45 pin inorganic membrane filter f) Reagents: Potassium phosphate monobasic (KH 2

PO

4 Phosphoric acid WO 2005/006871 PCT/US2004/020872 00 19 t Methanol

SCH

3

CN

Malic acid Standard ACS grade Citric acid Standard ACS grade \Oxalic acid Standard ACS grade 00 Quinic acid Standard ACS grade g) LC mobile phase: C1 0.2 M Phosphate buffer, pH 2.4: 00 SWeigh 27.2 g potassium dihydrogen phosphate in beaker. Add water to 950 mL. Using C, pH meter and 85% phosphoric acid, adjust to pH 2.4. Pour into 1 L graduate and adjust to volume with water; filter.

Preparation of Test Samples and Standards Working standard solutions.- Weigh 0.200 g each of ACS grade quinic, malic, citric, and oxalic acids to the nearest 0.1 mg. Dissolve in water, make up the volume to 100 mL in a volumetric flask with water and filter.

Test sample solutions.- Cartridge is conditioned by eluting 10 mL CH 3 CN -H 2 0 (50+50) through 10 mL Luer-Lok syringe. The syringe is removed and 10 mL air is passed through the cartridge. Ten mL of sample is eluted through the conditioned cartridge. First 4-5 mL is discarded and the next 4-5 mL is collected and filtered for LC analysis.

Determination: The system is conditioned with 100% methanol (or methanol-H 2 0[70+30]) followed by water and then phosphate buffer. The order is reversed at end of working day; never letting methyl alcohol to come in contact with phosphate buffer. Operating conditions: flow rate 0.80 mL/min; 214 nm detector; temperature ambient; sensitivity 0.1 AUFS.

Column system is satisfactory when baseline separation is achieved between sugar front peak and quinic acid in aqueous green tea extract. 5-20 pL standard solution is injected after each 2 test sample injections to check linearity. 5 L test sample solution is injected. An average of 2 injections are recorded for standard and test sample responses.

h) Calculations of Oxalic acid Oxalic acid (PA/PA') X X C where PA and PA' peak area of test sample and standard, respectively; V and V' volume of test sample and standard, respectively; and C concentration of standard, 9. Measurement of the Total Insoluble Calcium Components: WO 2005/006871 PCTIU S21)04/020872 00

O

O

4c This method is used to measure the level of total insoluble calcium components in consumable Sbeverages or aqueous tea extracts. Weigh 1000 g of the consumable beverage or 100 g of a w/w aqueous solution of green tea extract in a 2000 mL beaker. Add 20 mL of a calcium chloride solution. Allow to stand for 1 hour and then heat to 80 0 C and hold for \minutes. Filter through a tared filter paper. Wash the gelatinous precipitate with boiling water.

00 00 Then dry and weigh the precipitate as total insoluble calcium components.

O D. Examples 00 Example 1 A green tea fortified water beverage of the present invention is prepared according to the Sfollowing formulation.

Ingredient Wt. g/L Deionized Water 96.582 989.1 Magnesium Sulfate, anhydrous (Tangram Co.; Holtsville, NY)' 0.086 0.88 Vitamin Premix (Roche Vitamins, Inc.; Parsippany, New Jersey) 0.029 0.30 Sucrose, granulated (The Amalgamated Sugar Co.; Ogden, Utah) 2.682 27.47 Acesulfame Potassium (Nutrinova, Inc.; Somerset, New Jersey) 0.013 0.13 Sodium Citrate, dihydrate Staley; Decatur, Illinois) 0.155 1.59 Mono-potassium Phosphate (Astaris LLC; Carteret, New Jersey) 0.086 0.88 Green Tea Extract Powder (SynerTea™; Amax NutraSource, Inc.; 0.147 1.50 Eugene, Oregon) Citric Acid, anhydrous Staley; Decatur, Illinois) 0.220 2.25 TOTAL 100.0 1,024.1 Magnesium sulfate is 20.2% Mg 2 The vitamin premix provides the following nutrients per 250 mL serving of beverage: Vitamin C 22.2 mg Vitamin B12 0.79 tig Niacin 2.6 mg Vitamin E 3.96 International Units Vitamin B6 0.26 mg Zinc 1.74 mg The source of zinc in the vitamin premix is zinc gluconate (13.4% Zn).

The dry ingredients are weighed out and added to the deionized water with agitation. Mixing is continued until all the ingredients are in solution. The final beverage is clear with minimal brown color and no turbidity. The beverage has the following attributes: Green Tea Catechin Level (ppm mg/L) 400 ppm WO 2005/006871 PCT/US2004/020872 00 00

N

O

rsl Turbidity 22.3 NTU Absorbance at 430 nm (average of 2 measurements) 0.043 pH 3.61 Mineral Levels (per 250 mL beverage): Magnesium 44 mg 11.0% RDI Zinc 1.7mg= 11.3% RDI Carbohydrate Content (per 250 mL beverage) 7 g Calorie Content (per 250 mL beverage) 28 kcal The beverage is packaged in green colored plastic bottles. The in-package appearance is virtually identical to pure water.

Example 2 Two water beverage formulations fortified with green tea extract are prepared according to the following formulations: Formula A Wt. Grams/ Ingredient Liter Deionized Water 97.113 994.53 Magnesium Sulfate, anhydrous (Tangram Co.; Holtsville, NY)' 0.086 0.88 Zinc Gluconate (Glucona America, Inc.; Janesville, Wisconsin) 0.009 0.09 Sucrose, granulated (The Amalgamated Sugar Co.; Ogden, Utah) 2.682 27.47 Acesulfame Potassium (Nutrinova, Inc.; Somerset, New Jersey) 0.013 0.13 Green Tea Extract Powder (Product #285060; Plantextrakt, Inc., 0.098 1.00 Parsippany, New Jersey) TOTAL 100.0 1,024.1 Formula B Wt. Grams/ Ingredient Liter Deionized Water 96.652 989.81 Magnesium Sulfate, anhydrous (Tangram Co.; Holtsville, NY) 0.086 0.88 Zinc Gluconate (Glucona America, Inc.; Janesville, Wisconsin)2 0.009 0.09 Sucrose, granulated (The Amalgamated Sugar Co.; Ogden, Utah) 2.682 27.47 Acesulfame Potassium (Nutrinova, Inc.; Somerset, New Jersey) 0.013 0.13 Sodium Citrate, dihydrate Staley; Decatur, Illinois) 0.155 1.59 Mono-potassium Phosphate (Astaris LLC; Carteret, New Jersey) 0.086 0.88 WO 200(5/)006871 PCT/US2004/020872 Green Tea Extract Powder (Product #285060; Plantextrakt, Inc., 0.098 1.00 Parsippany, New Jersey) Citric Acid, anhydrous Staley; Decatur, Illinois) 0.220 2.25 TOTAL 100.0 1,024.1 1 Magnesium sulfate is 20.2% Mg 2 Zinc gluconate is 13.4% Zn Each beverage is prepared by weighing out the appropriate amount of dry ingredients and adding the dry ingredients to the deionized water with agitation. Mixing is continued until all the ingredients are in solution. The final beverages have the following attributes: Formula A Formula B Green Tea Catechin Level 150 ppm 150 ppm (ppm mg/L) Turbidity 15.3 NTU 4.1 NTU Absorbance at 430 nm 0.302 0.129 (average of 2 measurements) pH 5.22 3.60 Mineral Levels (per 250 mL): Magnesium 44 mg 11.0% RDI 44 mg= 11.0% RDI Zinc 3.0 mg 20% RDI 3.0 mg 20% RDI Formula B has a lower pH and yields a final beverage with a significantly lower absorbance at 430 nm and less visually apparent brown color, relative to formula A.

Example 3 A fortified sports drink of the present invention is prepared according to the following formulation.

Ingredient Wt. Grams/ Liter Deionized Water 93.279 955.27 Magnesium Sulfate, anhydrous (Tangram Co.; Holtsville, NY)' 0.154 1.58 Zinc Gluconate (Glucona America, Inc.; Janesville, Wisconsin) 2 0.009 0.09 Ferrous bis-glycinate (Ferrochel Albion Laboratories, Inc., 0.004 0.04 Clearfield, Utah) 3 Maltodextrin, 15 DE (Maltrin M150, Grain Processing Corp., 3.095 31.70 WO 2005/006871 PCT/US24104/020872 Muscatine, Iowa) Sucrose, granulated (The Amalgamated Sugar Co.; Ogden, Utah) 2.682 27.47 Acesulfame Potassium (Nutrinova, Inc.; Somerset, New Jersey) 0.013 0.13 Sodium Citrate, dihydrate Staley; Decatur, Illinois) 0.155 1.59 Sodium Chloride (IMC Salt, Inc.; Overland Park, Kansas) 0.033 0.34 Mono-potassium Phosphate (Astaris LLC; Carteret, New Jersey) 0.086 0.88 Green Tea Extract Powder (SynerTeaTM; Amax NutraSource, Inc.; 0.110 1.13 Eugene, Oregon) Citric Acid, anhydrous Staley; Decatur, Illinois) 0.244 2.50 FD&C Red No. 40 powder Goodrich, Cleveland, Ohio) 0.005 0.05 Natural Flavor (Mane, Inc., Milford, Ohio) 0.130 1.33 TOTAL 100.0 1,024.1 SMagnesium sulfate is 20.2% Mg 2 Zinc gluconate is 13.4% Zn 3 Ferrous bis-glycinate (Ferrochel®) is 20.2% Fe The dry ingredients are weighed out and added to the deionized water with agitation. Mixing is continued until all the ingredients are in solution. The natural flavor is then added with agitation.

The finished drink has the following attributes.

WO 2005/006871 PCT/US2004/020872 Green Tea Catechin Level (ppm mg/L) 300 ppm Turbidity 4.1 NTU pH 3.47 Mineral Levels (per 250 mL): Magnesium 80 mg 20% RDI Zinc 3.0 mg 20% RDI Iron 2.0mg 11% RDI Carbohydrate Content (per 250 mL beverage) 15.0 g Calorie Content (per 250 mL beverage) 60 kcal Example 4 A calcium compatible nano-filtered green tea extract is prepared as follows. To 100 g of a 5% w/w solution of SynerTea M green tea extract in water, 0.22 g of calcium sulfate dihydrate is added and stirred for 15 minutes. The treated extract is heated to 168 0 F (75.6 0 C) and held for minutes. The extract is then cooled and filtered through a 0.45 micron filter. The filtered extract thus obtained is free of insoluble calcium components, such as oxalic acid, and does not yield sedimentation on standing.

Before Treatment After Treatment Green Tea Extract Solution pH 5.24 5.03 Green Tea Catechin Level (g per 100 g on dry weight 2.56 2.50 basis) Insoluble Calcium Components (g per 100 g on dry 3.16 N.D.

weight basis) Oxalic Acid Content (g per 100 g on dry weight 2.62 <0.1 basis) N.D. not detected Example A calcium compatible green tea extract is prepared as follows. To 100 g of a 5% w/w solution of Plantextrakt (#285060) Green Tea extract in water, 1.6 g of calcium chloride dihydrate is added and stirred for 15 minutes. The treated extract is heated to 168 0 F (75.6 0 C) and held for minutes. The extract is then cooled and filtered through Whatman paper #1 and then through a WO 2005/006871 PCTUS20(041020872 0.45 micron filter. The filtered extract thus obtained is free of insoluble calcium components, such as oxalic acid and pectin, and does not yield sedimentation on standing.

Before Treatment After Treatment Green Tea Extract Solution pH 5.70 5.08 Green Tea Catechin Level (g per 100 g on dry weight 1.50 1.60 basis) Insoluble Calcium Components (g per 100 g on dry 4.94 N.D.

weight basis) Oxalic Acid Content (g per 100 g on dry weight 2.62 <0.1 basis) N.D. not detected Example 6 A water beverage fortified with calcium and green tea extract is prepared according to the following formulation. Prior to addition, an aqueous solution of the Plantextrakt green tea extract w/w) is treated to remove calcium insoluble components, such as oxalic acid and pectin, using the procedure described in Example 5 above.

Ingredient Wt. Grams/ Liter Deionized Water 93.309 955.58 Magnesium Sulfate, anhydrous (Tangram Co.; Holtsville, NY)' 0.086 0.88 Vitamin Premix (Roche Vitamins, Inc.; Parsippany, New Jersey) 2 0.029 0.30 Sucrose, granulated (The Amalgamated Sugar Co.; Ogden, Utah) 2.682 27.47 Acesulfame Potassium (Nutrinova, Inc.; Somerset, New Jersey) 0.013 0.13 Sodium Citrate, dihydrate Staley; Decatur, Illinois) 0.155 1.59 Mono-potassium Phosphate (Astaris LLC; Carteret, New Jersey) 0.086 0.88 Treated Green Tea Extract aqueous solution from Example 5 2.94 30.11 Citric Acid, anhydrous Staley; Decatur, Illinois) 0.220 2.25 Calcium gluconate (Glucona America, Inc.; Janesville, Wisconsin)3 0.480 4.92 TOTAL 100.0 1,024.1 Magnesium sulfate is 20.2% Mg 2 The vitamin premix provides the following nutrients per 250 mL serving of finished beverage: Vitamin C 22.2 mg Vitamin B12 0.79 pg Niacin 2.6 mg Vitamin E 3.96 International Units WO 2005/006871 PCT/US21)(4/020872 00 26 0 SVitamin B6 0.26 mg Zinc 1.74 mg SThe source of zinc in the vitamin premix is zinc gluconate (13.4% Zn).

Calcium gluconate is 9.31% Ca The dry ingredients are weighed out and added to the deionized water with agitation. The O) treated green tea extract solution from Example 5 is then added and mixing is continued until all C1 the ingredients are in solution. The final beverage is clear with no turbidity. The beverage has O the following attributes: 00 Green Tea Catechin Level (ppm mg/L) 227 ppm Turbidity 30 NTU pH 3.63 Mineral Levels (per 250 mL beverage): Magnesium 44 mg 11.0% RDI Zinc 1.7mg 11.3%RDI Calcium 115 mg 11.5% RDI Carbohydrate Content (per 250 mL beverage) 7 g Calorie Content (per 250 mL beverage) 28 kcal While particular embodiments of the present invention have been illustrated and described, it would be obvious to those skilled in the art that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.

Claims (20)

1. A process for treating a tea extract, the process comprising: providing a tea extract in a liquid form, combining the tea extract and a soluble calcium source, permitting the tea extract and soluble calcium source to interact under 00 conditions sufficient to form a precipitate of insoluble calcium components, and (N separating the insoluble calcium components from the tea extract. C 00

2. The process according to claim 1, wherein the soluble calcium source is added to the C tea extract at a concentration from about 400mg to about 10,000 mg per 100 gm of tea extract on a dry weight basis.

3. The process according to claim 1 or claim 2, wherein the soluble calcium source is selected form calcium sulfate, calcium chloride, calcium acetate, calcium gluconate, or mixtures thereof.

4. The process according to claim 3, wherein after treatment the insoluble calcium components in the tea extract on a dry weight basis is not present in more than about 1% w/w.

The process according to claim 3 or claim 4, wherein the insoluble calcium components in the tea extract on a dry weight basis is not present in more than 0.6% w/w.

6. The process according to any one of claims 1 to 5, wherein the insoluble calcium components in the tea extract on a dry weight basis is not present in more than 0.1% w/w.

7. The process according the any one of claims 1 to 6, wherein the tea extract at step (a) comprises at least 10% green tea catechins on a dry weight basis.

8. The process according to, any one of claims 1 to 7, wherein the tea extract at step (a) comprises at least 20% green tea catechins on a dry weight basis.

9. The process according to any one of claims 1 to 8, wherein treatment conditions at step are carried out at ambient temperatures. 28 00

10. The process according to any one of claims 1 to 9, wherein treatment conditions at Sstep are carried out at high temperature. t,

11. The process according to any one of claims 1 to 10, wherein the temperature is greater than about 130'F (54'C).

12. The process according to any one of claims 1 to 11, wherein the temperature is greater 00 than 145 0 F (63 0 C). CI 10

13. The process according to any one of claims 1 to 12, wherein the temperature is greater 00 than 160 0 F (71'C).

14. The process according to any one of claims 1 to 13, wherein treatment conditions at step are carried at a reaction time of 2 hours or less.

The process according to any one of claims 1 to 14, wherein treatment conditions at step are carried at a reaction time of 45 minutes or less.

16. The process according to any one of claims 1 to 15, wherein treatment conditions at step are carried at a reaction time of 15 minutes or less.

17. The process according to any one of claims 1 to 16, further comprising: addition of acid to reduce pH, thereby improving colour stability of the tea extract.

18. The process according to any one of claims 1 to 17, wherein the acid is a food-grade acid.

19. The tea extract treated by the process of any one of claims I to 18.

20. The process of any one of claims 1 to 18, substantially as herein before descried with reference to any of the Examples.