US 20110223256A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2011/0223256A1 Zhang et al. (43) Pub. Date: Sep. 15, 2011

(54) METHOD FOR STABILIZING FILAVONOID Publication Classification AQUEOUS DISPERSION (51) Int. Cl. A6IR 9/14 (2006.01) (75) Inventors: Naijie Zhang, Ridgefield, CT (US); A63L/352 (2006.01) Vincent E.A. Rinaldi, Bethel, CT A63L/335 (2006.01) (US); Peter S. Given, JR., A63L/05 (2006.01) Ridgefield, CT (US) A6II 3L/22 (2006.01) A63L/7048 (2006.01) (73) Assignee: STOKELY-VAN CAMP, INC., A6IP33/6 (2006.01) Chicago, IL (US) (52) U.S. Cl...... 424/493: 514/456; 514/450; 514/734; 514/679; 514/27 (21) Appl. No.: 13/036,161 (57) ABSTRACT A method for Suspending microparticulated water insoluble (22) Filed: Feb. 28, 2011 bioactive compound in a beverage by incorporating solubi lized or dispersed microparticulated compound and at least one dispersion stabilizer into a beverage. A composition com Related U.S. Application Data prising solubilized or dispersed microparticulated water (60) Provisional application No. 61/312,694, filed on Mar. insoluble bioactive compound and a dispersion stabilizer 11, 2010. agent.

E= 5. I Probe = 2f )A Aag = 50X Reference ag. = Curtiew. = r Feate = 24-327-2. late 14 Patent Application Publication Sep. 15, 2011 Sheet 1 of 4 US 2011/0223256 A1

&&. S SS E = i. Sigra A = WPSE G3 a = 9a urn I Prote = 201 A Aag = 5x Reference Aag = Cutiew, = . Fs are = 92-2-928-2 s. are: 2

SS = . Signal A = WPSG3 arrer = a Pa I Probe - 408 EA Aag = 6. KX Reference Aag = Out Dow. M = 5. Fis a = 92-928. F if date 52

Patent Application Publication Sep. 15, 2011 Sheet 3 of 4 US 2011/0223256 A1

gz'61-I Patent Application Publication Sep. 15, 2011 Sheet 4 of 4 US 2011/0223256 A1

NIGEHEJSEHNINE?NIHWNNILITE 9." 61-I

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METHOD FOR STABILIZING FILAVONOID BRIEF DESCRIPTION OF THE DRAWINGS AQUEOUS DISPERSION 0007 FIGS. 1A and 1B compare amorphous and crystalline quercetin, respectively. FIELD OF THE INVENTION 0008 FIGS. 2A and 2B show the results of a quercetin 0001. The present invention relates generally to a method stability study by FTIR and Carbon-13 NMR, respectively. for dispersing and Stabilizing water insoluble bioactive com 0009 FIG.3 shows samples of stable aqueous dispersions pounds in a liquid medium and a composition containing prepared in accordance with the present invention. and a Suspension agent. DETAILED DESCRIPTION OF THE INVENTION BACKGROUND 0010. The present invention relates to a method for stabi lizing Suspended microparticulated water insoluble bioactive 0002 Biologically active (bioactive) compounds such as compounds such as quercetin particles in a liquid medium, in general, are used as nutritional Supplements to Such as a beverage, and further to a composition containing provide, for example, antioxidants. Most of bioactive com solubilized or dispersed compound particles and at least one pounds are highly rigid and crystalline and water insoluble. dispersion stabilizer. Beverages, prepared with solubilized or Quercetin, in particular, is considered a powerful antioxidant. dispersed a bioactive compound and a dispersion stabilizer, A number of studies showed that quercetin is effective for the contain fine, stable dispersions. prevention of various diseases. 0011. The water insoluble bioactive compounds may be 0003 Flavonoids may be extracted from plants. For polyphenols, flavanoids, alkaloids, aldehyde, aryl ketone, example, quercetin is a natural, plant-derived, flavonoid. In benzofuranoid, benzopyranoid, diterpenoid, phenylpro particular, quercetin is the aglycone form of a number of other panoid, polyketide, sesquiterpenoid, monoterpenoid, and/or flavonoid glycosides, such as rutin and quercitrin, found in may be derived from plants, herbs, or botanicals. See for citrus fruit, cranberries, blueberries, buckwheat, onions, and example, Naturally Occurring Bioactive Compounds Edited other vegetables, fruits, and greenplants. The chemical struc by Mahendra Rai, Maria Cecillia Carpinella, 2006. Bioactive ture of quercetin is illustrated below: Compounds in Foods. Edited by John Gilbert and Hamide Z. Senyuva. Bioactive Compounds From Plants, Volume 154, John Wiley and Sons, 1990. OH 0012 Suitable polyphenols include quercetin, eriocitrin, OH neoeriocitrin, narirutin, naringin, hesperidin, hesperetin, neo hesperidin, neoponcirin, poncirin, rutin, isorhoifolin, rhoifo lin, , neodiosmin, , , tangeritin, HO O catechin, catechin gallate, epigallocatechin, epigallocatechin gallate, anthocyanin, heptamethoxyflavone, curcumin, res Veratrol, naringenin, tetramethoxyflavone, , and OH . OH O 0013 Suitable flavonoids and other bioactive compounds include quercetin, flavonones, , dihydroflavonols, flavonols, flavandiols, leucoanthocyanidins, flavonol glyco 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H sodes, flavonone glycosides, isoflavonoids, and neofla vonoids. In particular, the favonoids may be, but not limited chromen-4-one to, quercetin, eriocitrin, neoeriocitrin, narirutin, maringin, 0004. It is desired to use flavonoids as a nutritional supple hesperidin, hesperetin, neohesperidin, neoponcirin, poncirin, ment in food products such as beverages. Often such fla rutin, isorhoifolin, rhoifolin, dioSmin, neodiosmin, sinense vonoids are difficult to disperse and mix into beverage prod tin, nobiletin, tangeritin, catechin, catechin gallate, epigallo ucts at efficacious concentrations. Often such flavonoids will catechin, epigallocatechin gallate, oolong tea polymerized simply settle to the bottom of the container holding the bev polyphenol, anthocyanin, heptamethoxyflavone, daidzin, erage. For example, quercetin is typically obtained as a pow daidzein, biochaminn A, prunetin, genistin, glycitein, gly der and is insoluble in water. When added to liquid media, citin, genistein, 6.7.4 trihydroxyisoflavone, , , quercetin usually agglomerizes and settles to the bottom of , balcalein, apin, cupressuflavone, datiscetin, dios the beverage, thereby resulting in a product that is not visually metin, , , gossypetin, geraldol, hinokiflavone, appealing to the consumer. primuletin, pratol, , myricetin, , robinetin, 0005. Therefore, a need exists in the food and beverage quercetagetin, and hydroxy-4-flavone. industry to provide the consumer with a food product con 0014 Suitable compounds derived from plants, herbs, or taining bioactive compounds Such as quercetin wherein the botanicals include pyrrolizildine, alkaloids, and artemisinin. bioactive compound is stabilized in an aqueous Suspension. 0015 For ease of discussion, the application will be dis cussed in terms of quercetin. However, it is intended that the BRIEF SUMMARY process steps and compositions apply to all suitable bioactive compounds described above and below. 0006. The present invention relates to a method for stabi 0016. In one aspect of the invention, quercetin is solubi lizing Suspended quercetin or other Suspended water lized or dispersed prior to adding the quercetin to an aqueous insoluble bioactive compounds in a beverage and a composi Solution containing a dispersion stabilizer. tion for a beverage containing stabilized Suspended quercetin 0017. An aqueous dispersion of quercetin through solubi or other water insoluble bioactive compounds. lization reduces particle size, density, rigidity, and crystalline US 2011/0223256 A1 Sep. 15, 2011

structure resulting in increasing dispersion stability. SEM 0027 Subsequently, water is added to dilute the concen results show dispersed quercetin in gellan gum has amor trated dispersion to yield a stable quercetin dispersion having phous structure (FIG. 1A) compared to crystalline quercetin a concentration between 0.001-5.0 wt.%. The pH of the (FIG. 1B). diluted dispersion is adjusted to less than 7.0, typically 2.5 to 0018. The chemical stability of quercetin during heating solubilizing process in polyols is determined by analytical 6. characterization. Quercetin dispersion with or without gellan 0028. The at least one dispersion stabilizer can be a gum is isolated and purified from beverage. FIGS. 2A and 2B biopolymer or a modified polysaccharide Such as gellangum, show that the spectra of FT-IR and carbon-13 NMR, respec pectin, carrageenan, ghatti gum, acacia gum, guar gum, Xan tively, of quercetin dispersions through solubilization are than gum, locust gum, agar, starch, alginate, cellulose, pro identical with that of starting material quercetin. The results tein, hydrolyzed protein, modified starch, carboxyl methyl indicate that there is no structure change of quercetin during cellulose (CMC) or the combination thereof. heating solubilization process. 0029 Preferably the biopolymers are charged polymers 0019 Quercetin is solubilized by dissolving quercetin in Such as carboxyl-containing polymers and Sulfate-containing a) hot alcohol. Such as hot ethanol or polyols, orb) an alkaline polymers. It was discovered that anionic or cationic biopoly solution having a pH above 7 or c) a mixture of alcohol and mers such as pectin, gellan gum, carrageenan, gum arabic, alkaline Solution, to form solubilized quercetin Solution hav ghatti gum, CMC, whey protein isolate showed better disper ing 1 to 15 wt % quercetin. Suitable alcohols include, but not sion stability than non-ionic polymers. It is believed that limited to, ethanol, isopropyl alcohol, isobutyl alcohol, and benzyl alcohol, and polyols such as glycerol and propylene quercetin absorbed on a charged polymer exhibits stable glycol. Combinations of alcohols are also contemplated. aqueous dispersion due to electrostatic, steric repulsion Preferably ethanol is used in combination with an alkaline between the particles. There is no settling/aggregation after Solution of quercetin. stored at ambient conditions. The inventors discovered that 0020. When completely dissolved, the quercetin solution the length of time that the quercetin stays dispersed in the either in hot alcohol or alkaline is added into an aqueous liquid media varies depending on the type(s) of dispersion Solution containing a stabilizer with high shear mixing or stabilizer(s) used. For example, the quercetin can stay dis agitation. The resulting quercetin comes out of Solution persed in a beverage for about 12 hours to about six months or resulting in fine particle dispersion, Suspension. The alcohol longer, depending on the dispersion stabilizer(s) used. concentration is generally between 85-99 wt.%. 0030 Controlling pH during the process is critical to make 0021. In addition to solubilization, quercetin can be dis stable dispersions and to Suppress undesired color develop persed into the hot alcohol. Such as hot ethanol or polyol. ment. When an alkaline solution of quercetin is added into the resulting in alcohol dispersion slurry in order to reduce quer stabilizer-containing aqueous solution under high shear mix cetin particle size. Suitable alcohols include, but not limited ing, pH of the dispersion should be maintained below 6, to, ethanol, isopropyl alcohol, isobutyl alcohol, and benzyl typically below 5, or 2.5 to 4. Otherwise, if the dispersion pH alcohol, and polyols such as glycerol and propylene glycol. is alkaline, a dark brown color is developed and does not Combinations of alcohols are also contemplated. disappear even if the pH is lowered. 0022 Stable quercetin dispersion is obtained when hot alcohol dispersion slurry is added into an aqueous solution 0031. The quercetin is microparticulated. “Microparticu containing stabilizer under either high shear mixing or agita lated' or “microparticulate” as used in the instant application tion conditions. The concentration of quercetin dispersion means a small particle ranging in size from about 0.1 um to slurry in alcohol is in the range of 1-50 wt %. about 50 um with an average particle size below about 10um. 0023 The temperature of the hot alcohol for dissolving or 0032. The microparticulated quercetin has an average par dispersing the quercetin is between 50-200° C., generally ticle size less than 10 microns, in particular less than 3 65-170° C. microns or less than 1 micron. For example, at least 90% of 0024. The alkaline solution may be prepared from any the particles have a particle size less than 50 microns and 80% Suitable alkaline agent Such as sodium hydroxide or potas of the particles have a particle size less than 3 microns. sium hydroxide. The pH of the alkaline solution is above 7, 0033. The stability of quercetin aqueous dispersion is fur generally between 10 and 12. ther improved by adding an organic compound with density 0025. After the quercetin is solubilized or dispersed, the less than 1.0 grams per cubic centimeter into an alcohol quercetin is added to a solution containing at least one dis Solution during solubilization of quercetin. Organic com persion stabilizer. The solution is mixed or agitated to form pound can be flavor ingredient such as limonene, terpene, stable quercetin dispersion. In particular, the Solubilized or dispersed quercetin is added into a stabilizer-containing citral, ethylbutyrate, ethyl acetate or oil Such as coconut, aqueous solution under mixing/agitation to form a dispersion cotton seed, olive, corn, palm, peanut, rapeseed, safflower, containing from 0.1 to 10 weight% quercetin. The dispersion sesame, Soja beans, Sunflower, canola or combinations stabilizer is present in an amount Sufficient to Suspend the thereof. The concentration of organic compound in alcohol is microparticulated quercetin in the beverage. from 0.001 to 50 wt.%. 0026. The aqueous Solution containing dispersion stabi 0034. As noted, the description focused on quercetin for lizer is maintained at a temperature below 35° C. and at pH ease of discussion; however, other water insoluble bioactive less than 7, typically 3 to 5, during the addition of the solu compounds were also investigated using the present invention bilized alkaline quercetin solution. When quercetin is solubi dispersion method. Such bioactive compounds included cur lized or dispersed in hot alcohol, the solution or dispersion cumin, rutin, resveratrol, naringenin, hesperedin, tet slurry is cooled if necessary to between 50-130° C. prior to ramethoxyflavone (PMF), and artemisinin (anti malaria addition to stabilizer-containing aqueous Solution. drug). US 2011/0223256 A1 Sep. 15, 2011

-continued OCH CHO OCH OCH

O HO OH Curcumin CHO OCH OH O 3,6,3',4'-Tetramethoxyflavone

HO O

OH O Naringenin

OH H -C Artemisinin 0035. As set forth in the examples below, stable aqueous dispersions containing bioactive compounds were prepared by using the solubilization and stabilization process. The or resulting dispersions were fine, homogeneous, and stable. Resveratrol There were no settling, phase separation, and precipitates on the side of the bottle (See FIG. 3 and Table 1). The results indicate that the presented dispersion method could be widely OH used to make stable water insoluble compound aqueous dis persion in different application areas. HO O OH TABLE 1 OH HO Water-Insoluble Bioactive Compounds O O O H Aqueous Dispersion in 20 OZ Beverage OH O O Compound Amount (mg) Shelf Life (22-25° C.) Quercetin 1OOO Stable after at least 6 months H3 C O Curcumin 500 Stable after at least 3 months HO Rutin 500 Stable after at least 3 months HO Resveratrol 500 Stable after at least 3 months OH Naringenin 500 Stable after at least 3 months Rutin Hespered in 500 Stable after at least 3 months Tetramethoxyflavone 500 Stable after at least 3 months Artemisinin 100 Stable after at least 3 months HO HO 0036. The beverage may be any suitable beverage includ HO ing, but not limited to, juices, carbonated Soft drinks, water, dairy and isotonic beverages. One of ordinary skill in the art O of the chemical and food Sciences would recognize that any OH flavonoid may be used in accordance with the present inven HO O OCH tion. HO 0037 Applicants also discovered that although the bever O age appeared relatively viscous and/or thick upon adding the OH O quercetin (or other water-insoluble bioactive compounds) to the beverage, by using the solubilized/dispersed quercetin in conjunction with the dispersion stabilizer, as disclosed herein, the beverage had a thin consistency upon consump tion. This unexpected result, the thin consistency notwith OH O standing the viscous visual appearance, is also advantageous, Hespered in as it leads to consumer likability and acceptance of the bev erage. US 2011/0223256 A1 Sep. 15, 2011

0038 Food-grade preservatives and acidulants may also mabinlin, braZZein, hernandulcin, phyllodulcin, glycyphyl be added to the solubilized water-insoluble bioactive com lin, phloridzin, trilobtain, baiyunoside, osladin, polypodoside pounds. Incorporating Solubilized/dispersed water-insoluble A, pterocaryoside A, pterocaryoside B, mukurozioside, phlo bioactive compounds into the beverage results in improved misoside I, periandrin I, abrusoside A, and cyclocarioside I. suspension and dispersibility. It was further discovered that 0043 NHPS also includes modified NHPS's. Modified by incorporating the solubilized/dispersed water-insoluble NHPS's include NHPS's which have been altered naturally. bioactive compounds into the beverage, along with the dis For example, a modified NHPS includes, but is not limited to, persion stabilizer, that low concentration of the dispersion NHPS's which have been fermented, contacted with enzyme, stabilizer may be added with a high concentration of water or derivatized or substituted on the NHPS. In one embodi insoluble bioactive compounds, and even though the water ment, at least one modified NHPS may be used in combina insoluble bioactive compound is present in high concentra tion with at least one NHPS. In another embodiment, at least tions and the beverage may look Viscous, upon consumption one modified NHPS may be used without a NHPS. Thus, it tastes thin and is easily ingested by the consumer. modified NHPS's may be substituted for a NHPS or may be 0039. The method of the present invention may also used in combination with NHPS's for any of the embodiments include one or more additional ingredients selected from the described herein. For the sake of brevity, however, in the group consisting of carbohydrates, salts, salt blends, food description of embodiments of this invention, a modified grade acids, flavors, colors, Vitamin B3, Vitamin C, non NHPS is not expressly described as an alternative to an nutritive and/or nutritive Sweeteners and combinations of unmodified NHPS, but it should be understood that modified these ingredients. NHPS's can be substituted for NHPS's in any embodiment 0040 Sweeteners of beverage embodiments of the inven disclosed herein. tion include caloric carbohydrate Sweeteners, natural high 0044 As used herein, the phrase “synthetic sweetener potency Sweeteners, synthetic high-potency Sweeteners, refers to any composition that is not found in nature and is a other sweeteners, and combinations thereof. With the guid high potency Sweetener. Non-limiting examples of synthetic ance provided herein, a Suitable Sweetening system (whether sweeteners suitable for embodiments of this invention a single compound or combination thereof) can be selected. include Sucralose, acesulfame potassium (acesulfame K or 0041. Examples of suitable caloric carbohydrate sweeten aceK) or other salts, aspartame, alitame, saccharin, neohes ers include Sucrose, fructose, glucose, erythritol, maltitol, peridin dihydrochalcone, cyclamate, neotame, N-3-(3-hy lactitol, Sorbitol, mannitol. Xylitol, D-tagatose, trehalose, droxy-4-methoxyphenyl)propyl-L-C.-aspartyl-phenylala galactose, rhamnose, cyclodextrin (e.g., C.-cyclodextrin, nine 1-methyl ester, N-3-(3-hydroxy-4-methoxyphenyl)-3- B-cyclodextrin, and Y-cyclodextrin), ribulose, threose, arabi methylbutyl-L-C.-aspartyl-L-phenylalanine 1-methyl ester, nose, Xylose, lyxose, allose, altrose, mannose, idose, lactose, N-3-(3-methoxy-4-hydroxyphenyl)propyl-L-C.-aspartyl maltose, invert Sugar, isotrehalose, neotrehalose, palatinose L-phenylalanine 1-methyl ester, and salts thereof. or isomalitulose, erythrose, deoxyribose, gulose, idose, talose, 0045. The method described herein is advantageous as it erythrulose, Xylulose, psicose, turanose, cellobiose, glu Successfully Suspends water-insoluble bioactive compounds cosamine, mannosamine, fucose, glucuronic acid, gluconic Such as quercetin in the beverage and thereby deters settling acid, glucono-lactone, abequose, galactosamine, Xylo-oli of the water-insoluble bioactive compounds to the bottom of gosaccharides (Xylotriose, Xylobiose and the like), gentio the beverage's packaging. oligoscaccharides (gentiobiose, gentiotriose, gentiotetraose 0046. The present invention also relates to compositions and the like), galacto-oligosaccharides, Sorbose, nigero-oli comprising solubilized or dispersed water-insoluble bioac gosaccharides, fructooligosaccharides (kestose, nystose and tive compounds and at least one dispersion stabilizer. In one aspect of this invention, about 90% of the microparticulated the like), maltotetraol, maltotriol, malto-oligosaccharides water-insoluble bioactive compounds is below about 50 (maltotriose, maltotetraose, maltopentaose, maltohexaose, microns and the at least one dispersion stabilizer is present in maltoheptaose and the like), lactulose, melibiose, raffinose, an amount sufficient to suspend the water-insoluble bioactive rhamnose, ribose, isomerized liquid Sugars such as high fruc compounds in a liquid medium. tose corn/starch syrup (e.g., HFCS55, HFCS42, or HFCS90), 0047. The stability of water-insoluble bioactive com coupling Sugars, soybean oligosaccharides, and glucose pound aqueous dispersion is further improved by adding an syrup. organic compound with density less than 1.0 grams per cubic 0042. Other sweeteners suitable for use in embodiments centimeter into alcohol solution during solubilization of the provided herein include natural, synthetic, and other high water-insoluble bioactive compound. Suitable organic com potency Sweeteners. As used herein, the phrases “natural pounds can be flavor ingredient Such as limonene, terpene, high-potency sweetener.” “NHPS,” “NHPS composition.” citral, ethylbutyrate, ethyl acetate or oil Such as coconut, and “natural high-potency Sweetener composition' are syn cotton seed, olive, corn, palm, peanut, rapeseed, safflower, onymous. “NHPS' means any sweetener found in nature sesame, Soja beans, Sunflower, canola or the combination. which may be in raw, extracted, purified, treated enzymati The concentration of organic compound in alcohol is between cally, or any other form, singularly or in combination thereof O.1-30 wit. 9%. and characteristically has a Sweetness potency greater than 0048. The following examples are specific embodiments Sucrose, fructose, or glucose, yet has fewer calories. Non of the present invention, but are not intended to limit the limiting examples of NHPS's suitable for embodiments of invention. this invention include rebaudioside A, rebaudioside B, rebau dioside C (dulcoside B), rebaudioside D, rebaudioside E. Example 1 rebaudioside F, dulcoside A, rubusoside, Stevia, Stevioside, mogroside IV. mogroside V, Luo Han Guo Sweetener, Sia 0049. A quercetin solution was prepared by dissolving 1.0 menoside, monatin and its salts (monatin SS, RR, RS, SR), g quercetin to 20 g glycerol attemperature 150°C. Then the curculin, glycyrrhizic acid and its salts, thaumatin, monellin, solubilized quercetin Solution was slowly added to an aque US 2011/0223256 A1 Sep. 15, 2011

ous solution containing gellan gum under high mixing at pH added in the concentrations (w/w) listed below to make an 5. A homogeneous dispersion containing 0.5% quercetin was isotonic beverage containing quercetin. The pH range of the obtained. resultant isotonic beverage was 3.5. 0050. The concentrated quercetin dispersion was added to the beverage and Suspended in the beverage. Additional ingredients were added in the concentrations (w/w) listed below to make an isotonic beverage containing quercetin. The Amount pH was about 3.5. The pH range of the resultant isotonic Ingredient (% by wt.) beverage may be about 2.5-4.5. Water 92.16% Liquid Sucrose 2.963% Salt Blend O.176% Quercetin O.2% Gellangum O.O.3% Amount Citric Acid O.18.0% Ingredient (% by wt.) Glycerol 2% Propylene Glycol 2% Water 92.16% Mango Flavor O. 100% Liquid Sucrose 2.963% Yellow #6 Color 10% solution O.O60% Salt Blend O.176% Liq. Sucralose (25%) O.O2.1% Quercetin O.2% Gellangum O.O.3% Ace K O.OO3% Citric Acid O.18.0% Vitamin C (Ascorbic Acid) O. 105% Glycerol 4% Vitamin B3 (Niacinamide) O.004% Mango Flavor O.100% Total 100.000% Yellow #6 Color 10% solution O.O60% Liq. Sucralose (25%) O.O2.1% Ace K O.OO3% Vitamin C (Ascorbic Acid) O.105% Vitamin B3 (Niacinamide) O.004% Example 4 0053. The quercetin solution was prepared by dissolving Total 1OOOOO% 0.5 g quercetin into 10 g sodium hydroxide (0.5N) at room temperature. Then, quercetin alkaline solution was slowly added to an aqueous Solution containing carrageenan under Example 2 homogenization at pH 5. The dispersion pH was controlled 0051 Quercetin (1.0 g) was dissolved in 20 g propylene below 7 during addition of quercetin alkaline solution. A glycol at 150°C. Then, quercetin solution was slowly added homogeneous dispersion containing 0.5% quercetin is into gellan gum-containing aqueous solution under high obtained. The concentrated quercetin dispersion was added to shear mixing at pH 5. The quercetin dispersion was added to the beverage and Suspended in the beverage. beverage. Additional ingredients were added in the concen 0054 Additional ingredients, including a high intensity trations (w/w) listed below to make an isotonic beverage natural Sweetener were added to create a Zero calorie orange containing quercetin. The pH range of the resultant isotonic flavored isotonic beverage. The pH of the resultant beverage beverage was 3.5. was about 3.5.

Amount Amount Ingredient (% by wt.) Ingredient (% by wt.) Water 99.37% Water 92.16% Citric Acid O.OSO% Liquid Sucrose 2.963% Phosphoric Acid O.O16% Salt Blend O.176% Vitamin B3 O.005% Quercetin O.2% Ascorbic Acid O. 109% Gellangum O.O.3% Carrageenan O.1% Citric Acid O.18.0% Quercetin O.1% Propylene Glycol 4% Salt Blend O.176% Mango Flavor O.100% Reb A O.025% Yellow #6 Color 10% solution O.O60% Orange Flavor O.OSO% Liq. Sucralose (25%) O.O2.1% Ace K O.OO3% Total 100.000% Vitamin C (Ascorbic Acid) O.105% Vitamin B3 (Niacinamide) O.004% 0055 Alternatively, or in addition to, the salt blend listed Total 1OOOOO% above, the beverage may contain sea salt to produce a natural Zero-calorie isotonic beverage. Example 3 Example 5 0052 Quercetin (1.0 g) was dissolved in 10 g glycerol and 0056 Quercetin (0.5 g) was dispersed and partially solu 10 g propylene glycol at 150° C. Then, quercetin polyol bilized in 2.5g ethanol at 70° C. Then, the partially solubi Solution was slowly added into gellan gum-containing aque lized, dispersed quercetin slurry was slowly added into ghatti ous solution under high mixing at pH 5. The quercetin dis gum-containing aqueous solution under homogenization at persion was added to beverage. Additional ingredients were pH 5. The quercetin dispersion was added to beverage. Addi US 2011/0223256 A1 Sep. 15, 2011

tional ingredients were added in the concentrations (w/w) listed below to make an isotonic beverage containing querce tin. The pH was 3.5. Amount Ingredient (% by wt.) Water 94.17% Amount Liquid Sucrose 2.963% Ingredient (% by wt.) Salt Blend O.176% Quercetin O.1% Water 95.59% Gum Arabic O.1% Liquid Sucrose 2.963% Citric Acid O.18.0% Ethanol O.S Glycerol 2% Salt Blend O.176% limonene O.02% Quercetin O.1% Mango Flavor O. 100% Ghatti Gum O.2% Yellow #6 Color 10% solution O.O60% Citric Acid O.18.0% Liq. Sucralose (25%) O.O2.1% Mango Flavor O. 100% Ace K O.OO3% Yellow #6 Color 10% solution O.O60% Vitamin C (Ascorbic Acid) O. 105% Liq. Sucralose (25%) O.O2.1% Vitamin B3 (Niacinamide) O.004% Ace K O.OO3% Vitamin C (Ascorbic Acid) O. 105% Total 100.000% Vitamin B3 (Niacinamide) O.004% Total 100.000% Example 8 Example 6 0059. The quercetin solution was prepared by dissolving 0.5 g quercetin to 10 g glycerol and 1.0 g canola oil at 0057 Quercetin (0.5 g) was dissolved in 4 g propylene temperature 120°C. Then the solubilized quercetin solution glycol at 140°C. Then, the quercetin solution was slowly was slowly added to an aqueous solution containing modified added into gum arabic-containing aqueous Solution under starch (purity gum 2000) under homogenization at pH 5. A homogenization at pH 5. The quercetin dispersion was added homogeneous dispersion containing 0.5% quercetin was to beverage. Additional ingredients were added in the con obtained. The concentrated quercetin dispersion was added to centrations (w/w) listed below to make an isotonic beverage the beverage and Suspended in the beverage. Additional containing quercetin. The pH range of the resultant isotonic ingredients were added in the concentrations (w/w) listed beverage was 3.5. below to make an isotonic beverage containing quercetin. The pH was 3.5.

Amount Ingredient (% by wt.) Amount Water 95.29% Ingredient (% by wt.) Liquid Sucrose 2.963% Water 93.59% Salt Blend O.176% Liquid Sucrose 2.963% Quercetin O.1% Salt Blend O.176% Gum Arabic O.2% Propylene Glycol O.8% Quercetin O.1% Citric Acid O.18.0% Purity Gum 2000 O.S9/o Mango Flavor O.100% Citric Acid O.18.0% Yellow #6 Color 10% solution O.O60% Glycerol 2% Canola Oil O.2% Liq. Sucralose (25%) O.O2.1% Mango Flavor O. 100% Ace K O.OO3% Yellow #6 Color 10% solution O.O60% Vitamin C (Ascorbic Acid) O.105% Liq. Sucralose (25%) O.O2.1% Vitamin B3 (Niacinamide) O.004% Ace K O.OO3% Total 1OOOOO% Vitamin C (Ascorbic Acid) O. 105% Vitamin B3 (Niacinamide) O.004% Total 100.000% Example 7 0058. The quercetin solution was prepared by dissolving Example 9 0.5 g quercetin to 10 g glycerol and 0.1 g limonene at tem perature 120°C. Then the solubilized quercetin solution was 0060 Quercetin (0.5 g) was dissolved in 10 g glycerol at slowly added to an aqueous solution containing gum arabic 150° C. Then, the quercetin solution was slowly added into under homogenization at pH 5. A homogeneous dispersion pectin-containing aqueous solution under high shear mixing containing 0.5% quercetin was obtained. The concentrated at pH 5. The quercetin dispersion was added to beverage. quercetin dispersion was added to the beverage and Sus Additional ingredients were added in the concentrations pended in the beverage. Additional ingredients were added in (w/w) listed below to make an isotonic beverage containing the concentrations (w/w) listed below to make an isotonic quercetin. The pH range of the resultant isotonic beverage beverage containing quercetin. The pH was 3.0. was 3.5. US 2011/0223256 A1 Sep. 15, 2011

-continued

Amount Amount Ingredient (% by wt.) Ingredient (% by wt.) Water 94.09% Sodium Citrate O.13% Liquid Sucrose 2.963% Monopotassium Phosphate O.04% Salt Blend O.176% Citric Acid O.187% Quercetin O.1% Reb A O.O15% Pectin O.2% Erythritol O.885% Citric Acid O.18.0% Ascorbic Acid O.1.03% Glycerol 296 Gellan Gum O.O.3% Mango Flavor O.100% Glycerol 4% Yellow #6 Color 10% solution O.O60% Rutin O.0833% Liq. Sucralose (25%) O.O2.1% Sodium Benzoate O.1% Ace K O.OO3% Vitamin C (Ascorbic Acid) O.105% Total 100.000% Vitamin B3 (Niacinamide) O.004% Total 1OOOOO% Example 12 0063 Resveratrol (0.5 g) was dissolved in 24 g glycerol at Example 10 150° C. Then, resveratrol solution was slowly added into 0061 Curcumin (0.5 g) was dissolved in 24 g glycerol at gellan gum-containing aqueous solution under high shear 150° C. Then, curcumin solution was slowly added into gel mixing at pH 5. The resveratrol dispersion was added to langum-containing aqueous Solution under high shear mix beverage. Additional ingredients were added in the concen ing at pH 5. The curcumin dispersion was added to beverage. trations (w/w) listed below to make an isotonic beverage Additional ingredients were added in the concentrations containing resveratrol. The pH range of the resultant isotonic (w/w) listed below to make an isotonic beverage containing beverage was 3.5. curcumin. The pH range of the resultant isotonic beverage was 3.5. Amount Ingredient (% by wt.)

Amount Water 92.5% Ingredient (% by wt.) Sucrose 1.93% Sodium Citrate O.13% Water 92.5% Monopotassium Phosphate O.04% Sucrose 1.93% Citric Acid O.187% Sodium Citrate 0.13% Reb A O.O15% Monopotassium Phosphate O.04% Erythritol O.885% Citric Acid O.187% Ascorbic Acid O.1.03% Reb A O.O15% Gellan Gum O.O.3% Erythritol O.885% Glycerol 4% Ascorbic Acid O.1.03% Resveratrol O.0833% Gellan Gum O.O.3% Sodium Benzoate O.1% Glycerol 4% Curcumin O.0833% Total 100.000% Sodium Benzoate O.1%

Total 1OOOOO% Example 13 0064 Artemisinin (0.1 g) was dissolved in 24 g glycerol at Example 11 150° C. Then, artemisinin solution was slowly added into gellan gum-containing aqueous solution under high shear 0062 Rutin hydrate (0.5 g) was dissolved in 24 g glycerol mixing at pH 5. The artemisinin dispersion was added to at 150°C. Then, Rutin solution was slowly added into gellan beverage. Additional ingredients were added in the concen gum-containing aqueous solution under high shear mixing at trations (w/w) listed below to make an isotonic beverage pH 5. The Rutin dispersion was added to beverage. Additional containing artemisinin. The pH range of the resultant isotonic ingredients were added in the concentrations (w/w) listed beverage was 3.5. below to make an isotonic beverage containing Rutin. The pH range of the resultant isotonic beverage was 3.5.

Amount Ingredient (% by wt.) Amount Ingredient (% by wt.) Water 92.57% Sucrose 1.93% Water 92.5% Sodium Citrate O.13% Sucrose 1.93% Monopotassium Phosphate O.04% US 2011/0223256 A1 Sep. 15, 2011

-continued -continued

Amount Amount Ingredient (% by wt.) Ingredient (% by wt.) Citric Acid O.187% Erythritol O.885% Reb A O.O15% Ascorbic Acid O.1.03% Erythritol O.885% Gellan Gum O.O.3% Ascorbic Acid O.1.03% Glycerol 4% Gellan Gum O.O.3% Hesperedin O.0833% Glycerol 4% Sodium Benzoate O.1% Artemisinin O.017% Sodium Benzoate O.1% Total 100.000%

Total 1OOOOO% Example 16 Example 14 0067 3.6.3',4'-Tetramethoxyflavone (0.5 g) was dissolved in 24 g glycerol at 150° C. Then, tetramethoxyflavone solu 0065 Naringenin (0.5 g) was dissolved in 24 g glycerol at tion was slowly added into gellan gum-containing aqueous 150° C. Then, naringenin solution was slowly added into Solution under high shear mixing at pH 5. The tetramethoxy gellan gum-containing aqueous solution under high shear flavone dispersion was added to beverage. Additional ingre mixing at pH 5. The naringenin dispersion was added to dients were added in the concentrations (w/w) listed below to beverage. Additional ingredients were added in the concen make an isotonic beverage containing tetramethoxyflavone. trations (w/w) listed below to make an isotonic beverage The pH range of the resultant isotonic beverage was 3.5. containing naringenin. The pH range of the resultant isotonic beverage was 3.5.

Amount Ingredient (% by wt.) Amount Ingredient (% by wt.) Water 92.5% Sucrose 1.93% Water 92.5% Sodium Citrate O.13% Sucrose 1.93% Monopotassium Phosphate O.04% Sodium Citrate 0.13% Citric Acid O.187% Monopotassium Phosphate O.04% Reb A O.O15% Citric Acid O.187% Erythritol O.885% Reb A O.O15% Ascorbic Acid O.1.03% Erythritol O.885% Gellan Gum O.O.3% Ascorbic Acid O.1.03% Glycerol 4% Gellan Gum O.O.3% Tetramethoxyflavone O.0833% Glycerol 4% Sodium Benzoate O.1% Naringenin O.0833% Sodium Benzoate O.1% Total 100.000%

Total 1OOOOO% 0068. This invention may be embodied in other specific forms without departing from the spirit or essential charac teristics thereof. The foregoing embodiments, therefore, are Example 15 to be considered in all respects illustrative rather than limiting 0066 Hesperedin (0.5 g) was dissolved in 24 g glycerol at the invention described herein. The scope of the invention is 150° C. Then, hesperedin solution was slowly added into thus indicated by the appended claims, rather than by the gellan gum-containing aqueous solution under high shear foregoing description, and all changes that come within the mixing at pH 5. The hesperedin dispersion was added to meaning and range of equivalency of the claims are intended beverage. Additional ingredients were added in the concen to be embraced therein. trations (w/w) listed below to make an isotonic beverage We claim: containing hesperedin. The pH range of the resultant isotonic 1. A method for dispersing and Suspending microparticu beverage was 3.5. lated water insoluble bioactive compound particles in a bev erage comprising the steps of a) solubilizing water insoluble bioactive compound in a hot alcohol solution, or in an alkaline Solution, or combination thereof or b) dispersing the water Amount insoluble bioactive compound in a hot alcohol solution; and Ingredient (% by wt.) c) introducing the solubilized or dispersed bioactive com Water 92.5% pound into an aqueous solution containing at least one dis Sucrose 1.93% persion stabilizer; wherein the dispersion stabilizer is present Sodium Citrate 0.13% Monopotassium Phosphate O.04% in an amount Sufficient to Suspend the water insoluble bioac Citric Acid O.187% tive compound in the beverage, wherein the Suspended water Reb A O.O15% insoluble bioactive compound comprises microparticulated particles. US 2011/0223256 A1 Sep. 15, 2011

2. The method of claim 1 wherein the water insoluble 17. The method of claim 1 wherein the water insoluble bioactive compound is a polyphenol. bioactive compound is solubilized in an alkaline Solution at a 3. The method of claim 1 wherein the water insoluble pH of from 10 to 12. bioactive compound is a flavonoid. 18. The method of claim 1 wherein the alkaline solution 4. The method of claim3 wherein the flavonoid is selected comprises sodium hydroxide or potassium hydroxide. from the group consisting of quercetin, flavonones, flavones, 19. The method of claim 1 wherein the hot alcohol solution dihydroflavonols, flavonols, flavandiols, leucoanthocyani further comprises an organic compound selected from the dins, flavonol glycosodes, flavonone glycosides, isofla group consisting of limonene, terpene, citral, ethylbutyrate, vonoids, and neoflavonoids. ethyl acetate, coconut oil, cotton seed oil, olive oil, corn oil, 5. The method of claim 1 wherein the water insoluble palm oil, peanut oil, rapeseed oil, safflower oil, Sesame oil, bioactive compound is selected from the group consisting of Soja beans oil, Sunflower oil, canola oil, or combinations quercetin, flavonones, flavones, dihydroflavonols, flavonols, thereof. flavandiols, leucoanthocyanidins, flavonol glycosodes, fla 20. The method of claim 20 wherein the concentration of Vonone glycosides, isoflavonoids, and neoflavonoids. In par organic compound in alcohol is from 0.1 to 30 wt.%. ticular, the favonoids may be, but not limited to, quercetin, 21. The method of claim 1 wherein the beverage further eriocitrin, neoeriocitrin, narirutin, naringin, hesperidin, hes comprises at least one additional ingredient selected from the peretin, neohesperidin, neoponcirin, poncirin, rutin, isor group consisting of carbohydrates, salts, salt blends, food hoifolin, rhoifolin, dioSmin, neodiosmin, sinensetin, nobile grade acids, flavors, colors, Vitamin B3, Vitamin C, sweeten tin, tangeritin, catechin, catechin gallate, epigallocatechin, ers and combinations thereof. epigallocatechin gallate, oolong tea polymerized polyphenol, 22. The method of claim 22 wherein the beverage further anthocyanin, heptamethoxyflavone, daidzin, daidzein, bio comprises salt in a concentration of about 0.1-0.3 wt % based chaminn A, prunetin, genistin, glycitein, glycitin, genistein, on the total weight of the beverage and carbohydrate in a 6.7.4 trihydroxy isoflavone, morin, apigenin, Vitexin, balca concentration of about 1-3 wt % based on the total weight of lein, apin, cupressuflavone, datiscetin, , fisetin, the beverage. galangin, gossypetin, geraldol, hinokiflavone, primuletin, 23. The method of claim 22 wherein the beverage further pratol, luteolin, myricetin, orientin, robinetin, quercetagetin, comprises at least one Sweetener selected from the group and hydroxy-4-flavone. consisting of Stevia, monatin and combinations thereof. 6. The method of claim 1 wherein the water insoluble 24. The method of claim 1 wherein the hot alcohol is at a bioactive compound is extracted from plants, herbs, or temperature of 50 to 200° C. botanicals. 25. A beverage prepared in accordance with the method of claim 1. 7. The method of claim 1 wherein the water insoluble 26. A method for suspending flavonoid particles in a bev bioactive compound is selected from the group consisting of erage comprising combining at least one dispersion stabilizer curcumin, rutin, resveratrol, naringenin, hesperedin, tet and solubilized or dispersed microparticulated flavonoid, ramethoxyflavone (PMF), and artemisinin. wherein the dispersion stabilizer is present in an amount 8. The method of claim 1 wherein the water insoluble sufficient to suspend the microparticulated flavonoid in the bioactive compound is quercetin. beverage, and wherein the flavonoid is solubilized or dis 9. The method of claim 1 wherein the at least one disper persed in hot glycerol, hot propylene glycol, hot ethanol, an sion stabilizer is a n anionic or cationic biopolymer or a alkaline Solution, or combination thereof. modified polysaccharide selected from the group consisting 27. The method of claim 26 wherein the flavonoid is quer of gellan gum, pectin, guar gum, Xanthan gum, acacia gum, cetin. locust gum, agar, starch, ghatti gum, carrageenan, alginate, 28. The method of claim 26 wherein the at least one dis cellulose, protein, hydrolyzed protein, modified Starch, car persion stabilizer is an anionic or cationic biopolymer. boxyl methyl cellulose, or the combination thereof. 29. The method of claim 28 wherein the at least one dis 10. The method of claim 9 wherein the at least one disper persion stabilizer is a gellan gum, gum arabic, pectin, carra Sion stabilizer is a gellangum, pectin, gum arabic, ghattigum, geenan, ghatti gum, alginate, CMC, whey protein isolate, or carrageenan, alginate, CMC, whey protein isolate, or combi combination thereof. nation thereof. 30. The method of claim 26, wherein the particles are 11. The method of claim 1 wherein the dispersion stabilizer microparticulated. is present in the beverage in a concentration of about 0.001 31. A composition for a comestible comprising solubilized, 5.0 wt.% based on total weight of the beverage. dispersed microparticulated flavonoid and at least one disper 12. The method of claim 1 wherein the beverage has a pH sion agent, wherein about 90% of the flavonoid has a particle of less than 6. size below about 50 microns and the at least one dispersion 13. The method of claim 13 wherein the pH of the beverage agent is present in an amount Sufficient to suspend the fla is about 2.5 to about 4.5. vonoid in a liquid medium; wherein the microparticulated 14. The method of claim 1 wherein the alcohol is a polyol. flavonoid is solubilized or dispersed in hot glycerol, hot pro 15. The method of claim 1 wherein alcohol is selected from pylene glycol, hot ethanol, an alkaline Solution, or combina the group consisting of ethanol, benzyl alcohol, isopropyl tions thereof. alcohol, isobutyl alcohol, glycerol, and propylene glycol. 32. The composition for a comestible of claim 30 wherein 16. The method of claim 1 wherein the water insoluble the flavonoid is quercetin. bioactive compound is solubilized in a hot glycerol or hot propylene glycol or hot ethanol. c c c c c