Novel mogrosides and their uses

Abstract

Provided herein are mogrosides, including non-glucose glycosides. Also provided are compositions, including consumables, that contain the novel mogrosides described herein.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 810,556, filed February 26, 2019, which is incorporated by reference in its entirety.

[0002] FIELD OF THE INVENTION The present disclosure relates to mogroside compounds containing one or more non-glucose carbohydrates and methods for making the same. More particularly, the present disclosure relates to the use of mogroside compounds containing one or more non-glucose carbohydrates as sweeteners in consumable products. [Background technology]

[0003] background Monk fruit extracts, obtained from Siraitia grosvenori (a plant in the Cucurbitaceae family), are commercially used as natural sweeteners. However, monk fruit extracts can have taste characteristics that discourage their use as a substitute for high-calorie sweeteners (e.g., sugar) in food and beverage compositions. For example, the extracts may have a particular off-flavor or lingering aftertaste, or the sweetness may take longer than desired to develop after ingestion (i.e., delayed onset of sweetness).

[0004] There remains a need for sweeteners with reduced calorie content, and foods and beverages containing same, that have low or no calories and improved taste characteristics. Summary of the Invention [Means for solving the problem]

[0005] overview Typical mogrosides are characterized by glucose-containing glycosides at the C-3 and C-24 positions of the mogrol core. The present disclosure relates to novel mogroside compounds that contain at least one carbohydrate other than glucose in the glycoside at the C-3 and C-24 positions.

[0006] In one aspect, the present invention provides a compound of formula I: [ka] (In the formula, [ka] is a double bond, then X is O; [ka] is a single bond, X is selected from OH and H, and the carbon to which X is attached is replaced by H to provide the correct valence (e.g., HC[(CH2)(C)]-X); R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are each independently selected from hydrogen or a pentose or hexose carbohydrate, where R 1 ~R 6 at least one of which is a pentose or hexose carbohydrate that is not glucose; and The total number of carbohydrates is 4, 5, or 6. of mogrosides.

[0007] In each case, the pentose or hexose carbohydrate can be selected from any known pentose or hexose carbohydrate. Exemplary pentose and hexose carbohydrates include, but are not limited to, glucose, xylose, ribose, arabinose, deoxyglucose, kyxose, lyxose, fucose, allose, allulose, altrose, mannose, gulose, idose, galactose, talose, and rhamnose. The carbohydrate bond can be in the α- or β-configuration. The carbohydrate can be in the D- or L-configuration.

[0008] In one embodiment, R 1 ~R 4 One of the carbohydrates is a pentose or hexose carbohydrate that is not glucose, and R 5 and R 6 is hydrogen.

[0009] In another embodiment, R 1 ~R 4 One of them is xylose, and R 5 and R 6 is hydrogen.

[0010] In certain embodiments, the total number of carbohydrates in the compound of Formula I is 4, 5, or 6, and the non-glucose pentose or hexose carbohydrate is xylose.

[0011] In certain other embodiments, the total number of carbohydrates in the compound of Formula I is 4, 5, or 6, and the non-glucose pentose or hexose carbohydrate is not xylose.

[0012] In a second aspect, the present invention provides a compound of formula II: [ka] (In the ceremony [ka] is a double bond, then X is O; [ka] is a single bond, X is selected from OH and H, and the carbon to which X is attached is replaced by H to provide the correct valence (e.g., HC[(CH2)(C)]-X); R 1 , R 2 , R 3 , R 4 , and R 7 are each independently selected from hydrogen or a pentose or hexose carbohydrate, where R 1 , R 2 , R 3 , R 4 , and R 7 at least one of which is a pentose or hexose carbohydrate other than glucose) of mogrosides.

[0013] In each case, the pentose or hexose carbohydrate can be selected from any known pentose or hexose carbohydrate. Exemplary pentose and hexose carbohydrates include, but are not limited to, glucose, xylose, ribose, arabinose, deoxyglucose, chrysose, lyxose, fucose, allose, allulose, altrose, mannose, gulose, idose, galactose, talose, and rhamnose. The carbohydrate linkage can be in the α- or β-configuration.

[0014] In certain embodiments, when the total number of carbohydrates in the compound of Formula II is 3, 4, 5, or 6, the non-glucose pentose or hexose carbohydrate is xylose.

[0015] Specific mogrosides of the present disclosure include mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranoside}, mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-L-rhamnopyranosyl-(1→6)]-β-D-glucopyranoside}, mogrol-3-O-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside]-24-O -[β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside], Mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-D-galactopyranosyl-(1→6)]-β-D-glucopyranoside}, Mogrol-3-O-[α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside], Mogrol-3-O-[{β-D-glucopyranosyl-(1→6)}-β -D-glucopyranoside]-24-O-[{β-D-xylopyranosyl-(1→2)}-{β-D-glucopyranosyl-(1→6)}-β-D-glucopyranoside], Mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside, Mogrol-3-O-{[β-D-xylopyranosyl-(1→4)]-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[β-D-gluco pyranosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}, mogrol-3-O-{[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[α-L-rhamnopyranosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}, mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside],Mogrol-3-O-[β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside], Mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-β-D-glucopyranoside, 3-O-β-D-glucopyranosylmogrol 24-O-α-L-rhamnopyranosyl -(1→2)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside, and mogrol-3-O-{[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranoside},

[0016] In certain embodiments, the mogrosides are isolated and purified.

[0017] In a further aspect, the invention is a composition comprising at least one mogroside described herein. In certain embodiments, the invention is a composition comprising at least one isolated and purified mogroside described herein.

[0018] In one embodiment, the present invention is a sweetener composition comprising at least one mogroside described herein.

[0019] In another embodiment, the present invention is a taste enhancing composition comprising at least one mogroside described herein, wherein when added to a consumable product, the mogroside is present in the composition in an amount effective to provide a concentration below the flavor recognition threshold of the mogroside.

[0020] In yet another embodiment, the invention is a sweetener enhancer composition comprising at least one mogroside described herein, wherein the mogroside is present in the composition in an amount effective to provide a concentration below the sweetness perception threshold of the mogroside when the sweetener enhancer composition is added to a consumable product.

[0021] In yet another embodiment, the present invention is a consumable comprising at least one mogroside described herein. Suitable consumables include, but are not limited to, liquid-based or dry consumables, such as pharmaceutical compositions, edible gel mixes and compositions, dental compositions, food products, beverages, and drink products.

[0022] In certain embodiments, the invention is a beverage comprising at least one mogroside described herein, hi certain embodiments, the mogroside is present in the beverage at a concentration at, above, or below the threshold sweetness perception of the mogroside.

[0023] In another aspect, the invention is a method of preparing a consumable product, comprising: (i) providing a consumable matrix; and (ii) adding at least one mogroside described herein to the consumable matrix to provide a consumable product.

[0024] In certain embodiments, the present invention is a method of preparing a beverage comprising: (i) providing a beverage matrix; and (ii) adding at least one mogroside described herein to the beverage matrix to provide a beverage.

[0025] In another aspect, the present invention is a method for enhancing the sweetness of a consumable, comprising: (i) providing a consumable comprising at least one sweetening component; and (ii) adding at least one isolated and purified mogroside described herein to the consumable to provide an enhanced sweetness consumable, wherein the mogroside is present in the enhanced sweetness consumable at a concentration below the sweetness perception threshold of the mogroside. In certain embodiments, the consumable is a beverage.

[0026] In some embodiments, the compositions of the present invention comprise one or more sweeteners, additives, and / or functional ingredients.

[0027] In one embodiment, the invention is a consumable product comprising at least one mogroside of the invention and one or more sweeteners, additives, and / or functional ingredients. In another embodiment, the invention is a beverage comprising at least one mogroside of the invention formula and one or more sweeteners, additives, and / or functional ingredients.

[0028] In another aspect, the present invention is a method for purifying a mogroside of the invention, comprising: (i) passing a solution containing a raw material comprising a mogroside of the invention through an HPLC column; and (ii) eluting fractions containing a mogroside of the invention to provide a purified mogroside composition comprising at least about 80% by weight of a mogroside of the invention. Exemplary raw materials include, but are not limited to, mixtures of mogrosides, luo han guo extracts (commercially available or prepared), and compositions resulting from the bioconversion processes described herein. [Brief explanation of the drawings]

[0029] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] FIG. 1 shows the LC-MS of 90% mogroside V starting material (MV90) (Example 1). [Figure 2] 1 shows an HPLC trace of the reaction of MogIIIE with rhamnose (Example 2). [Figure 3A] 1 shows the Maldi mass spectrum of peak 45 isolated from the reaction of MogIIIE with rhamnose (Example 2). [Figure 3B] 1 shows the Maldi mass spectrum of peak 46 isolated from the reaction of MogIIIE with rhamnose (Example 2). [Figure 4] 1 shows an HPLC trace of the reaction of MogIIIE with galactose (Example 2). [Figure 5A] 1 shows the Maldi mass spectrum of peak 108 isolated from the reaction of MogIIIE with galactose (Example 2). [Figure 5B]1 shows the Maldi mass spectrum of peak 109 isolated from the reaction of MogIIIE with galactose (Example 2). [Figure 6] 1 shows an MS trace of the reaction of MogIIIE with xylose (Example 2). [Figure 7] 1 shows the Maldi mass spectrum of peak 39 isolated from the reaction of MogIIIE with xylose (Example 2). DETAILED DESCRIPTION OF THE INVENTION

[0030] Detailed Description Disclosed herein are compounds, compositions, and methods for use as sweeteners in beverages and foods. The disclosed compounds include mogroside compounds containing non-glucose carbohydrate units. Surprisingly, it has been found that the mogrosides of the present invention are useful in a variety of applications, for example, as sweeteners or food additives in consumable products such as beverages and foods. Such compositions include the mogrosides of the present invention, alone or mixed with other sweeteners, additives, or taste enhancers.

[0031] I. Definition As used herein, the term "consumable" refers to a substance suitable for consumption by an individual. Exemplary consumables include, but are not limited to, edible gel mixes and compositions, dental compositions, food products (confectionery, seasonings, chewing gum, cereal compositions, baked goods, dairy products, and tabletop sweetener compositions), beverages and beverage products.

[0032] As used herein, the term "monk fruit" or "luo han guo" refers to the fruit of Siraitia grosvenori, a member of the Cucurbitaceae family.

[0033] As used herein, the term "pH" refers to a number that represents the acidity or alkalinity of a solution on a logarithmic scale, where 7 is neutral, lower values are more acidic, and higher values are more alkaline. pH is equal to -log10c, where c is the hydrogen ion concentration in moles / liter.

[0034] As used herein, the term "purified" means that the purity of a compound has been increased such that the compound is present in a more pure form than it is in its natural environment and / or extract. Purity is a relative term and does not necessarily imply absolute purity.

[0035] II. Compounds The present disclosure provides compounds of formula I: [ka] (In the ceremony [ka] is a double bond, then X is O; [ka] is a single bond, X is selected from OH and H, and the carbon to which X is attached is replaced by H to provide the correct valence (e.g., HC[(CH2)(C)]-X); R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 are each independently selected from hydrogen or a pentose or hexose carbohydrate, where R 1 ~R 6 at least one of which is a pentose or hexose carbohydrate that is not glucose; and The total number of carbohydrates is 4, 5, or 6. The present invention provides a mogroside compound of the formula:

[0036] In each case, the pentose or hexose carbohydrate can be selected from any known pentose or hexose carbohydrate. Exemplary pentose and hexose carbohydrates include, but are not limited to, glucose, xylose, ribose, arabinose, deoxyglucose, lyxose, fucose, allose, allulose, altrose, mannose, gulose, idose, galactose, talose, and rhamnose. The carbohydrate bond can be in the α- or β-configuration. The carbohydrate can be in the D- or L-configuration.

[0037] The compound of Formula I has at least four total carbohydrates (i.e., the sum of the carbohydrate units at the C-3 and C-24 positions). In one embodiment, the compound of Formula I has four total carbohydrates. In yet another embodiment, the compound of Formula I has five total carbohydrates. In yet another embodiment, the compound of Formula I has six total carbohydrates.

[0038] In one embodiment, R 1 ~R 4 One of the carbohydrates is a pentose or hexose carbohydrate that is not glucose, and R 5 and R 6 is hydrogen.

[0039] In one embodiment, the total number of carbohydrates in the compound of formula I is 4, 5, or 6, and the non-glucose pentose or hexose carbohydrate is xylose.

[0040] In another embodiment, the total number of carbohydrates in the compound of formula I is 4, 5, or 6, and the non-glucose pentose or hexose carbohydrate is rhamnose.

[0041] In yet another embodiment, the total number of carbohydrates in the compound of formula I is 4, 5, or 6, and the non-glucose pentose or hexose carbohydrate is galactose.

[0042] In certain embodiments, the mogroside of formula I is of formula Ia: [ka] (In the formula, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 is as defined above for formula I) It is the 11-oxomogroside of

[0043] In certain other embodiments, the mogroside of formula I is of formula Ib: [ka] (In the formula, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 is as defined above for formula I) It is 11-deoxomogroside of

[0044] In certain other embodiments, the mogroside of formula I is of formula Ic: [ka] (In the formula, R 1 , R 2 , R 3 , R 4 , R 5 , and R 6 is as defined above for formula I) is a compound of

[0045] The 11-OH can be in the R or S configuration.

[0046] The present disclosure provides compounds of formula II: [ka] (In the ceremony [ka] is a double bond, then X is O; [ka] is a single bond, X is selected from OH and H, and the carbon to which X is attached is replaced by H to provide the correct valence (e.g., HC[(CH2)(C)]-X); R 1 , R 2 , R 3 , R 4 , and R 7 are each independently selected from hydrogen or a pentose or hexose carbohydrate, where R 1 , R 2 , R 3 , R 4 , and R 7 at least one of which is a pentose or hexose carbohydrate other than glucose) It also provides mogrosides.

[0047] In each case, the pentose or hexose carbohydrate can be selected from any known pentose or hexose carbohydrate. Exemplary pentose and hexose carbohydrates include, but are not limited to, glucose, xylose, ribose, arabinose, deoxyglucose, chrysose, lyxose, fucose, allose, allulose, altrose, mannose, gulose, idose, galactose, talose, and rhamnose. The carbohydrate linkage can be in the α- or β-configuration.

[0048] The compound of Formula II has at least three total carbohydrates (i.e., the sum of the carbohydrate units at the C-3 and C-24 positions). In one embodiment, the compound of Formula II has three total carbohydrates. In another embodiment, the compound of Formula II has four total carbohydrates. In yet another embodiment, the compound of Formula II has five total carbohydrates. In yet another embodiment, the compound of Formula II has six total carbohydrates.

[0049] In one embodiment, the total number of carbohydrates in the compound of formula II is 3, 4, 5, or 6, and the non-glucose pentose or hexose carbohydrate is xylose.

[0050] In another embodiment, the total number of carbohydrates in the compound of formula II is 3, 4, 5, or 6, and the non-glucose pentose or hexose carbohydrate is rhamnose.

[0051] In certain embodiments, the mogroside of formula II is represented by formula IIa: [ka] (In the formula, R 1 , R 2 , R 3 , R 4 , and R 7 is as defined above for formula II) It is the 11-oxomogroside of

[0052] In certain other embodiments, the mogroside of formula II is of formula IIb: [ka] (In the formula, R 1 , R 2 , R 3 , R 4 , and R 7 is as defined above for formula II) It is 11-deoxomogroside of

[0053] In certain other embodiments, the mogroside of formula II is of formula IIc: [ka] (In the formula, R 1 , R 2 , R 3 , R 4 , and R 7 is as defined above for formula II) is a compound of

[0054] The 11-OH can be in the R or S configuration.

[0055] In one embodiment, the mogroside of the present invention is selected from: (i) Mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranoside} (CC-00489): [ka] (ii) Mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-L-rhamnopyranosyl-(1→6)]-β-D-glucopyranoside} (CC-00491): [ka] (iii) Mogrol-3-O-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside] (CC-00497): [ka] (iv) Mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-D-galactopyranosyl-(1→6)]-β-D-glucopyranoside} (CC-00498): [ka] (v) Mogrol-3-O-[α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside] (CC-00500): [ka] (vi) Mogrol-3-O-[{β-D-glucopyranosyl-(1→6)}-β-D-glucopyranoside]-24-O-[{β-D-xylopyranosyl-(1→2)}-{β-D-glucopyranosyl-(1→6)}-β-D-glucopyranoside] (CC-00507): [ka] (vii) Mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside] (CC-00518) [ka] (viii) Mogrol-3-O-{[β-D-xylopyranosyl-(1→4)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside} (CC-00520) [ka] (ix) Mogrol-3-O-{[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[α-L-rhamnopyranosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside} (CC-00539) [ka] (x) Mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside] (CC-00540) [ka] (xi) Mogrol-3-O-[β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside] (CC-00541) [ka] (xii) Mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-β-D-glucopyranoside (CC-00542) [ka] (xiii) 3-O-β-D-Glucopyranosylmogrol 24-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (CC-00550) [ka] (xiv) Mogrol-3-O-{[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranoside} (CC-00551) [ka]

[0056] In an exemplary embodiment, the mogrosides of the invention are isolated and purified.

[0057] As used herein, the term "isolated and purified" means that the compound is about 95% or greater by weight, i.e., greater than 95%, pure on a dry basis. The remainder of the mixture is typically other mogrosides and / or luo han guo extract. In more specific embodiments, the mogrosides of the formulas described herein have a purity of about 96% or greater, about 97% or greater, about 98% or greater, or about 99% or greater. In some embodiments, the mogrosides are produced enzymatically.

[0058] In some embodiments, the mogrosides of the present invention are sweet. The sweetness of a given composition is typically measured relative to a solution of sucrose. See generally, "A Systematic Study of Concentration-Response Relationships of Sweeteners," G.E. DuBois, D.E. Walters, S.S. Schiffman, Z.S. Warwick, B.J. Booth, S.D. Pecore, K. Gibes, B.T. Carr, and L.M. Brands, in Sweeteners: Discovery, Molecular Design and Chemoreception, D.E. Walters, F.T. Orthoefer, and G.E. DuBois, Eds., American Chemical Society, Washington, DC (1991), pp. 261-276.

[0059] The sweetness of a non-sucrose sweetener can be measured against a sucrose standard by determining the non-sucrose sweetener's sucrose equivalent (SE). Typically, taste panelists are trained to detect the sweetness of a standard sucrose solution containing 1-15% sucrose (w / v). Other non-sucrose sweeteners are then tasted at a series of dilutions to determine the concentration of the non-sucrose sweetener that is as sweet as a given percentage sucrose standard. For example, if a 1% solution of a non-sucrose sweetener is as sweet as a 10% sucrose solution, the sweetener is said to be 10 times more potent than sucrose and has a 10% sucrose equivalent.

[0060] In one embodiment, the mogroside, when added to the consumable, is present in an amount that provides a sucrose equivalent of greater than about 2% (w / v), such as, for example, greater than about 3% SE, about 4% SE, about 5% SE, about 6% SE, about 7% SE, about 8% SE, about 9% SE, about 10% SE, about 11% SE, about 12%, about 13% SE, or about 14% SE.

[0061] The amount of sucrose in a reference solution, and therefore another measure of sweetness, can be described in degrees Brix (°Bx). 1 degree Brix is 1 gram of sucrose in 100 grams of solution and expresses the strength of the solution as a percentage by weight (strictly speaking, by mass) (% w / w). In one embodiment, the mogrosides of the present invention, when added to a consumable, are present in an amount that provides a sweetness equivalent of about 0.50 to 14 degrees Brix, e.g., about 5 to about 12 degrees Brix, about 7 to 10 degrees Brix, or greater than 10 degrees Brix.

[0062] In exemplary embodiments, the isolated and purified mogrosides of the present invention have a sweetness that is about 30% or greater, such as about 40% or greater, about 50% or greater, about 60% or greater, about 70% or greater, about 80% or greater, or about 90% or greater, compared to partially purified mogrosides or luo han guo.

[0063] In other exemplary embodiments, the isolated and purified mogrosides of the invention have at least about 30% less bitterness (a taste stimulated by certain substances such as quinine, caffeine, and sucrose octaacetate), e.g., at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, compared to partially purified mogrosides or luo han guo. In certain embodiments, the isolated and purified mogrosides of the invention have substantially no bitterness. Methods for measuring the bitterness of a compound are known in the art.

[0064] In yet other exemplary embodiments, the isolated and purified mogrosides of the invention have a lingering sweet aftertaste (sweetness intensity after expectoration) that is at least about 30% less, e.g., at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, compared to partially purified mogrosides or luo han guo. In certain embodiments, the isolated and purified mogrosides of the invention have substantially no lingering sweet aftertaste. Methods for measuring lingering sweet aftertaste are known in the art.

[0065] In yet other exemplary embodiments, the isolated and purified mogrosides of the invention have at least about 30% less metallic taste (a taste associated with metal, tin, or iron), e.g., at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%, compared to partially purified mogrosides or luo han guo. In certain embodiments, the isolated and purified mogrosides of the invention have substantially no metallic taste.

[0066] In exemplary embodiments, the isolated and purified mogrosides of the present invention exhibit a maximal response (maximal sweetness achieved with increasing compound concentration (%SE)) that is at least about 30% higher, e.g., at least about 40% higher, at least about 50% higher, at least about 60% higher, at least about 70% higher, at least about 80% higher, or at least about 90% higher, compared to partially purified mogrosides or luohanguo. Methods for measuring the maximal response of a compound are known in the art. In one embodiment, the method is an in vitro cellular assay. In some embodiments, the cells express a sweet taste receptor or a sweet taste receptor dimer.

[0067] In other exemplary embodiments, the isolated and purified mogrosides of the present invention exhibit a sweet taste onset (time to maximum sweetness experience) that is at least about 30% shorter, e.g., at least about 40% shorter, at least about 50% shorter, at least about 60% shorter, at least about 70% shorter, at least about 80% shorter, or at least about 90% shorter, compared to partially purified mogrosides or luohanguo. Methods for measuring sweet taste onset are known in the art. In one embodiment, the method is an in vitro cellular assay. In some embodiments, the cells express a sweet taste receptor or a dimer of a sweet taste receptor.

[0068] III. Composition The present invention includes compositions comprising at least one mogroside of the present invention. "Composition," as that term is used herein, refers to a mixture of at least one mogroside of the present invention with at least one other substance.

[0069] In certain embodiments, the at least one other substance is not naturally occurring with the diterpene glycoside and / or is not naturally mixed with the diterpene glycoside, and therefore, these compositions are not naturally occurring.

[0070] In one embodiment, the invention is a composition comprising at least one mogroside of the invention provided as part of a mixture. In certain embodiments, the mixture is selected from the group consisting of mogrosides, luohanguo, by-products of other mogroside isolation and purification processes, commercially available mogroside extracts, by-products of biotransformation reactions, or any combination thereof.

[0071] In one embodiment, the mixture comprises at least one mogroside of the present invention in an amount ranging from about 1% to about 99% by weight on a dry basis, such as, for example, about 5% to about 99%, about 10% to about 99%, about 20% to about 99%, about 30% to about 99%, about 40% to about 99%, about 50% to about 99%, about 60% to about 99%, about 70% to about 99%, about 80% to about 99%, and about 90% to about 99%. In certain embodiments, the mixture comprises at least one mogroside of the present invention in an amount greater than about 90% by weight on a dry basis, such as, for example, greater than about 91%, greater than about 92%, greater than about 93%, greater than about 94%, greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, and greater than about 99%.

[0072] The composition may comprise at least about 5% by weight of a mogroside of the invention, such as, for example, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 97%.

[0073] In other embodiments, the composition has a total mogroside content of about 95% or greater by weight on a dry basis. In some embodiments, the composition has a total mogroside content of about 96% or greater, about 97% or greater, about 98% or greater, or about 99% or greater. As used herein, "total mogroside content" refers to the sum of the relative weight contributions of each mogroside in a sample, including the non-glucose-substituted mogrosides described herein.

[0074] As used herein, the term "purified mogroside" refers to mogrosides present in a mixture, e.g., luohanguo, at least about 50% by weight, such as, for example, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 97%.

[0075] In one embodiment, the present invention is a composition comprising at least one mogroside described herein provided as a pure compound, i.e., greater than 99% by weight on a dry basis.

[0076] The mogrosides of the present invention may be present in the composition in an amount effective to provide a concentration of the mogrosides of the present invention of about 1 ppm to about 10,000 ppm, when the composition is added to a consumable product, such as, for example, about 1 ppm to about 4,000 ppm, about 1 ppm to about 3,000 ppm, about 1 ppm to about 2,000 ppm, about 1 ppm to about 1,000 ppm, about 1 ppm to about 600 ppm, about 1 ppm to about 500 ppm, about 1 ppm to about 400 ppm, about 1 ppm to about 300 ppm, about 1 ppm to about 200 ppm, or about 1 ppm to about 100 ppm.

[0077] In another embodiment, the mogroside of the present invention is present in the composition in an amount effective to provide a concentration of the mogroside of the present invention of from about 10 ppm to about 1,000 ppm, from about 25 ppm to about 1,000 ppm, from about 50 ppm to about 1,000 ppm, or from about 100 ppm to about 1,000 ppm.

[0078] The weight ratio of the at least one other substance to the mogroside of the present invention can vary. Typically, the weight ratio of the at least one other substance to the mogroside of the present invention is, for example, from about 100:1 to about 2:1, from about 50:1 to about 2:1, from about 25:1 to about 2:1, from about 10:1 to about 2:1, from about 5:1 to about 2:1, from about 500:1 to about 400:1, from about 500:1 to about 300:1, from about 500:1 to about 200:1, or from about 500:1 to about 100:1. , about 500:1 to about 50:1, about 500:1 to about 25:1, about 500:1 to about 10:1, about 400:1 to about 300:1, about 400:1 to about 200:1, about 400:1 to about 100:1, about 400:1 to about 50:1, about 400:1 to about 25:1, about 400:1 to about 10:1, about 400:1 to about 6.67:1, about 300:1 to about 200: 1, about 300:1 to about 100:1, about 300:1 to about 50:1, about 300:1 to about 25:1, about 300:1 to about 10:1, about 300:1 to about 6.67:2, about 200:1 to about 100:1, about 200:1 to about 50:1, about 200:1 to about 25:1, about 200:1 to about 10:1, about 100:1 to about 50:1, about 100:1 to about 25:1 , about 500:1 to about 2:1, such as about 100:1 to about 10:1, about 100:1 to about 6.67:1, about 50:1 to about 25:1, about 50:1 to about 25:1, about 50:1 to about 10:1, about 50:1 to about 6.65:1, about 25:1 to about 10:1, about 25:1 to about 6.67:1, about 10:1 to about 6.67:1, and any range therebetween.

[0079] A. Sweetener Composition As noted above, in some embodiments, the mogrosides of the present invention are sweet. Accordingly, the present invention also provides sweetener compositions comprising at least one mogroside of the present invention. "Sweetener composition," as that term is used herein, refers to a mixture of at least one mogroside of the present invention and at least one other substance.

[0080] In certain embodiments, the at least one other substance is not naturally occurring with the mogroside and / or is not naturally mixed with the mogroside. Thus, these sweetener compositions are not naturally occurring. In one embodiment, the at least one other substance modulates the taste profile of the at least one mogroside to provide a composition having a more sucrose-like taste profile compared to the naturally occurring mogroside and (if applicable) the at least one other naturally occurring substance. For example, in certain embodiments, the composition exhibits one or more of the following properties: improved sweetness potency, improved texture, reduced sweetness linger, reduced bitterness, and / or reduced metallic taste.

[0081] In one embodiment, the sweetener composition comprises a mogroside of the present invention in a sweetening amount. As used herein, "sweetening amount" refers to the amount of a compound required to provide a detectable sweetness when present in a consumable product, such as a beverage, also referred to as the "sweetness perception threshold."

[0082] In one embodiment, the sweetener composition, when added to a sweetened composition or sweetened consumable, provides more than about 2% (w / v) sucrose equivalent, such as, for example, more than about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, or about 14%.

[0083] In certain embodiments, the sweetener composition comprises at least about 5% by weight of a mogroside of the present invention, such as, for example, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 97%.

[0084] In certain embodiments, the sweetener composition comprises at least about 5% by weight of at least one other substance (e.g., a sweetener), e.g., at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 97%.

[0085] In some embodiments, the at least one other substance is a sweetener, i.e., the sweetener composition is or includes a sweetener blend. Such a sweetener composition, when added to a sweetened composition or sweetened consumable, may provide more than about 2% (w / v) sucrose equivalent, such as, for example, more than about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, or about 14%.

[0086] In one embodiment, the at least one other substance is a sweetener that is not the same as a mogroside of the present invention. The sweetener may be present in a sweetening amount.

[0087] In one embodiment, at least one other substance is a natural high-potency sweetener.As used herein, the phrase "natural high-potency sweetener" refers to any sweetener that can be found in plants or other species without further modification, and has a higher sweetening potency than sucrose, fructose or glucose, but has lower calories.Natural high-potency sweeteners can be provided as pure compounds or as part of extracts.

[0088] In another embodiment, the at least one other substance is a synthetic sweetener. As used herein, the phrase "synthetic sweetener" refers to any composition that is not naturally occurring in nature and that characteristically has a higher sweetening potency than sucrose, fructose, or glucose but fewer calories.

[0089] In yet other embodiments, combinations of natural high-potency sweeteners and synthetic sweeteners are contemplated.

[0090] In another embodiment, the at least one other substance is a carbohydrate sweetener, such as, but not limited to, sucrose, glyceraldehyde, dihydroxyacetone, erythrose, threose, erythrulose, arabinose, lyxose, ribose, xylose, ribulose, xylulose, allose, altrose, galactose, glucose, gulose, idose, mannose, talose, fructose, psicose, sorbose, tagatose, mannoheptulose, sedoheltulose, octolose, fucose, rhamnose, arabinose, turanose, sialose, and combinations thereof.

[0091] Other suitable sweeteners include rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, rebaudioside I, rebaudioside H, rebaudioside L, rebaudioside K, rebaudioside J, rebaudioside M, rebaudioside N, rebaudioside O, dulcoside A, dulcoside B, rubusoside, stevia, stevioside, mogroside IV, mogroside V, mogroside VI, luohanguo, siamenoside I, mogroside IIIE, monatin and its salts (monatin SS, RR, RS, SR), curculin, glycyrrhizic acid and its salts, thaumatin, monellin, mabinlin, brazzein, hernandulcine, and the like. Examples of suitable steviol glycosides include steviol glycosides, phyllodulcin, glycyphyllin, phlorizin, trilobatin, bayounoside, osladin, polypodoside A, pterocaryoside A, pterocaryoside B, mukurozioside, phlomisoside I, periandrin I, abrusoside A, steviolbioside, hesperitin and cyclocaryoside I, sugar alcohols such as erythritol, sucralose, acesulfame potassium, acesulfamic acid and its salts, aspartame, alitame, saccharin and its salts, neohesperidin dihydrochalcone, cyclamate, cyclamic acid and its salts, neotame, advantame, glucosylated steviol glycosides (GSG), and combinations thereof.

[0092] In certain embodiments, the sweetener is at least one calorie-providing carbohydrate sweetener.

[0093] In one embodiment, the sweetener is a caloric sweetener or a mixture of caloric sweeteners, hi another embodiment, the caloric sweetener is selected from sucrose, fructose, glucose, high fructose corn / starch syrup, beet sugar, cane sugar, and combinations thereof.

[0094] In another embodiment, the sweetener is a rare sugar selected from allulose, sorbose, lyxose, ribulose, xylose, xylulose, D-allose, L-ribose, D-tagatose, L-glucose, L-fucose, L-arabinose, turanose, kojibiose, and combinations thereof.

[0095] In one embodiment, the sweetener composition includes at least one additional mogroside. The mogroside can be provided in pure form or as part of a mixture, i.e., a mogroside blend. Exemplary mogrosides include grosmogroside I, mogroside IA, mogroside IE, 11-oxomogroside IA, mogroside II, mogroside IIA, mogroside IIB, mogroside II E, 7-oxomogroside IIE, mogroside III, mogroside IIIE, 11-oxomogroside IIIE, 11-deoxymogroside III, mogroside IV, and mogroside IVA. Examples of the hydroxybenzoates include, but are not limited to, 11-oxomogroside IV, 11-oxomogroside IVA, mogroside V, isomogroside V, 11-oxoshiamenoside I, 11-deoxymogroside V, 7-oxomogroside V, 11-oxomogroside V, isomogroside V, mogroside VI, mogrol, 11-oxomogrol, siamenoside I, 11-oxosiamenosdie I, isomers of siamenoside I (e.g., those disclosed in WO20170119032; incorporated herein by reference in its entirety), particularly any of the 1,6-α isomers of siamenoside I, and combinations thereof. Additional exemplary mogrosides include those described in U.S. Patent Application Publication No. 2016039864, the contents of which are incorporated herein by reference.

[0096] In one embodiment, the at least one additional substance is siamenoside I in a sweetening amount.

[0097] In another embodiment, the at least one additional substance is a sweetening amount of the 1,6-α isomer of siamenoside I (mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}).

[0098] In yet another embodiment, the at least one additional substance is mogroside V in a sweetening amount.

[0099] In yet another embodiment, the at least one additional substance is rebaudioside M in a sweetening amount.

[0100] In yet another embodiment, the at least one additional substance is rebaudioside A in a sweetening amount.

[0101] In a further embodiment, the at least one additional substance is rebaudioside D in a sweetening amount.

[0102] IV. Taste-Enhancing Compositions In some embodiments, the mogrosides of the present invention are taste enhancers. For example, in some embodiments, at least one mogroside of the present invention modulates one or more taste attributes of a consumable sweetened with a non-sucrose sweetener, thereby making the consumable taste more similar to a sucrose-sweetened consumable. Exemplary taste attribute modulations include increased sweetness, reduced or eliminated bitterness, reduced or eliminated bitter linger, reduced or eliminated sourness, reduced or eliminated astringency, reduced or eliminated saltiness, reduced or eliminated metallic notes, improved texture, reduced or eliminated sweet linger, and increased sweetness expression. Multiple taste attributes of a sweetener can be simultaneously modulated so that the consumable has more of an overall sucrose-sweetened character. Methods for quantifying the improvement in sucrose-sweetened character are known in the art, such as taste testing and histogram mapping.

[0103] In certain embodiments, the mogrosides of the present invention are sweetness enhancers or modulators. "Sweetness enhancer," as that term is used herein, refers to a compound that enhances, amplifies, or intensifies the perception of sweetness in a consumable (e.g., a beverage) when the compound is present in the consumable at a concentration below the compound's sweetener recognition threshold, i.e., at a concentration at which the compound does not contribute to a noticeable sweetness in the absence of added sweeteners.

[0104] "Sweetness modulating agent," as that term is used herein, refers to a compound that, when present in a consumable (e.g., a beverage) at a concentration below the compound's sweetness perception threshold, alters the sweetness taste characteristics (e.g., lingering taste, or off-notes) of the consumable.

[0105] The term "sweetness potentiator" is synonymous with the terms "sweet taste potentiator," "sweetness potentiator," "sweetness amplifier," and "sweetness intensifier."

[0106] In one embodiment, the mogrosides of the present invention can be added directly, i.e., as compounds rather than provided in the form of a composition, to a consumable to enhance its sweetness. In this embodiment, the mogrosides of the present invention are added to the consumable at a concentration below their sweetness perception threshold, i.e., as sweetness enhancers. In certain embodiments, the mogrosides of the present invention are added to the consumable at a concentration below their sweetness perception threshold, i.e., as sweetness enhancers.

[0107] In certain embodiments, the mogrosides of the present invention are sweetness enhancers or modulators and are added to the consumable in an amount that will provide a concentration of mogroside that is at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% or more below its sweetness perception threshold.

[0108] In some embodiments, the mogrosides of the present invention increase the sucrose equivalent (SE) of the consumable by at least about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 1.5%, about 2.0%, about 2.5%, about 3.0%, about 4.0%, or about 5.0% compared to the SE of the consumable in the absence of the mogrosides of the present invention.

[0109] In other embodiments, at least one mogroside of the present invention may be added to a consumable in the form of a sweetener composition. A "sweetener composition," as that term is used herein, refers to a composition of the present invention described above that, when added to a consumable (e.g., a beverage), enhances, amplifies, or intensifies the sweetness perception of the consumable when the mogroside of the present invention is present in the sweetener composition in an amount that would provide a concentration of the mogroside that is below its sweetness perception threshold. In certain embodiments, the mogroside of the present invention is present in an amount that would provide a concentration of the mogroside of the present invention that is below its sweetness perception threshold.

[0110] It is contemplated that the sweetness enhancer composition may contain one or more sweetness enhancers or modulators in addition to at least one mogroside of the present invention. In one embodiment, the sweetness enhancer composition may contain one additional sweetness enhancer. In other embodiments, the composition may contain two or more additional sweetness enhancers. In embodiments in which two or more sweetness enhancers or modulators are utilized, each must be present at or below its respective sweetness perception threshold concentration.

[0111] The one or more other sweetness enhancers or modulators may be 2-hydroxybenzoic acid, 3-hydroxybenzoic acid, 4-hydroxybenzoic acid, 2,4-dihydroxybenzoic acid, 3,4-dihydroxybenzoic acid, 2,5-dihydroxybenzoic acid, 2,6-dihydroxybenzoic acid, 2,3,4-trihydroxybenzoic acid, 2,4,6-trihydroxybenzoic acid, 3-aminobenzoic acid, 4-aminobenzoic acid, 4-O-β-D-glucosyl-hesperetin dihydrochalcone, MG isomogrosaide V, 4-hydroxycinnamic acid, 4-methoxycinnamic acid, 1-(2-hydroxybenzoic acid), ... (2,4-Dihydroxyphenyl)-3-(4-pyridyl)-1-propanone, 4-ethoxybenzonitrile, 2-Methoxy-5-(phenoxymethyl)-phenol, 1-(2,4-dihydroxyphenyl)-2-(3-methoxy-4-hydroxyphenyl)-ethanone, Hesperetin Dihydrochalcone-4'-β-D-glucoside, Hesperetin, 2,3',6-Trihydroxy-4'-methoxydihydrochalcone, N-(3'-Methoxy-4'-hydroxybenzyl)-2,4,6-trihydroxybenzamide, 3'-7-Dihydroxy-4'-methoxyflavan, Phloretin, FEMA The antioxidants may be selected from the group consisting of, but not limited to, GRAS flavor 4669, FEMA GRAS flavor 4701, FEMA GRAS flavor 4720, FEMA GRAS flavor 4774, FEMA GRAS flavor 4708, FEMA GRAS flavor 4728, FEMA GRAS flavor 4601, FEMA GRAS flavor 4802, hesperitin dihydrochalcone, FEMA GRAS flavor 4872, FEMA GRAS flavor 4899, 4-amino-5-(cyclohexyloxy)-2-methylquinoline-3-carboxylic acid, rebaudioside M, rebaudioside N, rebaudioside O, rebaudioside C, and combinations thereof.

[0112] In another specific embodiment, the mogrosides of the present invention are flavor enhancers. "Flavor enhancer," as that term is used herein, refers to a compound that enhances, amplifies, or intensifies the perception of a flavor component when present in a consumable (e.g., a beverage) at a concentration below the compound's flavor recognition threshold, i.e., at a concentration at which the compound does not contribute to a discernible flavor in the absence of any flavor component (i.e., any substance that imparts sweetness, sourness, saltiness, savory, bitterness, metallic taste, etc.). The term "flavor recognition threshold," as used generally herein, is the lowest known concentration of a compound that is detectable as a particular flavor by the human palate. The flavor recognition threshold concentration is specific to a particular compound and may vary with temperature, matrix, ingredients, and / or flavor system.

[0113] The term "flavor enhancer" is synonymous with the terms "flavor potentiator," "flavor amplifier," and "flavor intensifier."

[0114] In one embodiment, at least one mogroside of the present invention is added to a consumable directly, i.e., as a compound rather than provided in the form of a composition, to enhance the flavor. In this embodiment, the mogroside of the present invention is added to the consumable at a concentration below its flavor recognition threshold, i.e., is a flavor enhancer. In certain embodiments, the mogroside of the present invention is added to the consumable at a concentration below its flavor recognition threshold, i.e., is a flavor enhancer.

[0115] The mogrosides of the present invention enhance the flavor of the consumable by at least about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1.0%, about 1.5%, about 2.0%, about 2.5%, about 3.0%, about 4.0%, or about 5.0% compared to the flavor of the consumable in the absence of the mogrosides of the present invention.

[0116] In other embodiments, at least one mogroside of the present invention may be added to a consumable in the form of a flavor enhancer composition. A "flavor enhancer composition," as that term is used herein, refers to a mixture of at least one mogroside of the present invention and at least one flavor component, wherein the at least one mogroside is mixed with the at least one flavor component, and when added to a consumable (e.g., a beverage), the composition enhances, amplifies, or intensifies the perception of the flavor component in the consumable when the at least one mogroside of the present invention is present in the flavor enhancer composition in an amount that would provide a concentration of the mogroside that is below its flavor recognition threshold.

[0117] The addition of the flavor enhancer composition increases the detected flavor of at least one flavor component in the consumable compared to the detected flavor of the same component in the consumable in the absence of the flavor enhancer. Without being bound by theory, because the flavor enhancer is present in the consumable at a concentration below its flavor recognition threshold, the flavor enhancer composition likely does not impart a discernible taste to the consumable to which it is added.

[0118] Suitable flavoring ingredients include, but are not limited to, vanillin, vanilla extract, mango extract, cinnamon, citrus, coconut, ginger, viridiflorol, almond, menthol (including mint-free menthol), grape skin extract, and grape seed extract. "Flavoring agent" and "flavoring ingredient" are synonymous and may include natural or synthetic substances or combinations thereof. Flavoring agents also include any other substance that imparts flavor and may include natural or non-natural (synthetic) substances that are safe for humans or animals when used within generally acceptable limits. Non-limiting examples of proprietary flavoring agents include Doehler™ Natural Flavoring Sweetness Enhancer K14323 (Doehler™, Darmstadt, Germany), Symrise™ Natural Flavor Mask for Sweeteners 161453 and 164126 (Symrise™, Holzminden, Germany), Natural Advantage™ Bitterness Blockers 1, 2, 9, and 10 (Natural Advantage™, Freehold, New Jersey, USA), and Sucramask™ (Creative Research Management, Stockton, California, USA).

[0119] In another embodiment, a flavor enhancing composition comprising at least one mogroside of the present invention, when added to a consumable product, enhances flavors (individual flavors or overall flavors), including, but not limited to, fruit flavors, including tropical fruit flavors, and vanilla-caramel type flavors.

[0120] V. Additives In another aspect, the compositions described herein (i.e., the compositions described above and the consumables described below) may include one or more additional additives and / or functional ingredients.

[0121] Exemplary additives include, but are not limited to, carbohydrates, polyols, amino acids and their corresponding salts, polyamino acids and their corresponding salts, sugar acids and their corresponding salts, nucleotides, organic acids, inorganic acids, organic salts including organic acid salts and organic base salts, inorganic salts, bitter compounds, caffeine, flavoring agents and flavoring ingredients, astringent compounds, proteins or protein hydrolysates, surfactants, emulsifiers, plant extracts, flavonoids, alcohols, polymers, and combinations thereof.

[0122] In one embodiment, the composition further comprises one or more polyols. As used herein, the term "polyol" refers to a molecule containing two or more hydroxyl groups. Polyols can be diols, triols, or tetraols, containing two, three, and four hydroxyl groups, respectively. Polyols can also contain five or more hydroxyl groups, such as pentaols, hexaols, or heptaols, containing five, six, or seven hydroxyl groups, respectively. Furthermore, polyols can be sugar alcohols, polyhydric alcohols, or polyalcohols, which are reduced forms of carbohydrates in which the carbonyl group (aldehyde or ketone, reducing sugar) has been reduced to a primary or secondary hydroxyl group.

[0123] Non-limiting examples of polyols in some embodiments include maltitol, mannitol, sorbitol, lactitol, xylitol, isomalt, propylene glycol, glycerol (glycerin), threitol, galactitol, palatinose, reduced isomaltooligosaccharides, reduced xylooligosaccharides, reduced gentiooligosaccharides, reduced maltose syrup, reduced glucose syrup, and sugar alcohols, or any other carbohydrate that can be reduced without adversely affecting taste.

[0124] Suitable amino acid additives include, but are not limited to, aspartic acid, arginine, glycine, glutamic acid, proline, threonine, theanine, cysteine, cystine, alanine, valine, tyrosine, leucine, arabinose, trans-4-hydroxyproline, isoleucine, asparagine, serine, lysine, histidine, ornithine, methionine, carnitine, aminobutyric acid (α-, β-, and / or δ-isomers), glutamine, hydroxyproline, taurine, norvaline, sarcosine, and salt forms thereof, such as sodium or potassium salts or acid salts. Amino acid additives may be in the D- or L-configuration and may be mono-, di-, or tri-forms of the same or different amino acids. Furthermore, amino acids may be α-, β-, γ-, and / or δ-isomers, where appropriate. Combinations of the aforementioned amino acids and their corresponding salts (e.g., their sodium, potassium, calcium, magnesium salts, or other alkali or alkaline earth metal salts, or acid salts) are also suitable additives in some embodiments. Amino acids may be natural or synthetic. Amino acids may be modified. A modified amino acid refers to any amino acid in which at least one atom has been added, removed, substituted, or a combination thereof (e.g., an N-alkyl amino acid, an N-acyl amino acid, or an N-methyl amino acid). Non-limiting examples of modified amino acids include amino acid derivatives such as trimethylglycine, N-methyl-glycine, and N-methyl-alanine. As used herein, modified amino acids encompass both modified and unmodified amino acids. As used herein, amino acids also encompass both peptides and polypeptides (e.g., dipeptides, tripeptides, tetrapeptides, and pentapeptides) such as glutathione and L-alanyl-L-glutamine.Suitable polyamino acid additives include poly-L-aspartic acid, poly-L-lysine (e.g., poly-L-α-lysine or poly-L-ε-lysine), poly-L-ornithine (e.g., poly-L-α-ornithine or poly-L-ε-ornithine), poly-L-arginine, other polymeric forms of amino acids, and their salt forms (e.g., calcium, potassium, sodium, or magnesium salts, such as L-glutamic acid monosodium salt). The polyamino acid additives may be in the D- or L-configuration. Furthermore, the polyamino acids may be α-, β-, γ-, δ-, and ε-isomers, as appropriate. Combinations of the foregoing polyamino acids and their corresponding salts (e.g., their sodium, potassium, calcium, magnesium salts, or other alkali or alkaline earth metal salts, or acid salts) are also suitable additives in some embodiments. The polyamino acids described herein may also include copolymers of different amino acids. The polyamino acids may be natural or synthetic. Polyamino acids may be modified such that at least one atom is added, removed, substituted, or a combination thereof (e.g., N-alkyl polyamino acids or N-acyl polyamino acids). As used herein, polyamino acids encompass both modified and unmodified polyamino acids. For example, modified polyamino acids include, but are not limited to, polyamino acids of various molecular weights (MW), such as poly-L-α-lysine with a MW of 1,500, 6,000, 25,200, 63,000, 83,000, or 300,000.

[0125] Suitable sugar acid additives include, but are not limited to, aldonic acid, uronic acid, aldaric acid, alginic acid, gluconic acid, glucuronic acid, glucaric acid, galactaric acid, galacturonic acid, and salts thereof (e.g., sodium, potassium, calcium, magnesium salts, or other physiologically acceptable salts), and combinations thereof.

[0126] Suitable nucleotide additives include, but are not limited to, inosine monophosphate ("IMP"), guanosine monophosphate ("GMP"), adenosine monophosphate ("AMP"), cytosine monophosphate (CMP), uracil monophosphate (UMP), inosine diphosphate, guanosine diphosphate, adenosine diphosphate, cytosine diphosphate, uracil diphosphate, inosine triphosphate, guanosine triphosphate, adenosine triphosphate, cytosine triphosphate, uracil triphosphate, alkali or alkaline earth metal salts thereof, and combinations thereof. The nucleotides described herein may also include nucleotide-related additives, such as nucleosides or nucleic acid bases (e.g., guanine, cytosine, adenine, thymine, uracil).

[0127] Suitable organic acid additives include any compound containing a -COOH moiety, such as C2-C30 carboxylic acids, substituted hydroxyl C2-C30 carboxylic acids, butyric acid (ethyl ester), substituted butyric acid (ethyl ester), benzoic acid, substituted benzoic acids (e.g., 2,4-dihydroxybenzoic acid), substituted cinnamic acids, hydroxy acids, substituted hydroxybenzoic acids, anisic acid, substituted cyclohexylcarboxylic acids, tannic acid, aconitic acid, lactic acid, tartaric acid, citric acid, isocitric acid, gluconic acid, glucoheptonic acid, adipic acid, hydroxycitric acid, malic acid, fruit acids, and the like. Acids (a blend of malic, fumaric, and tartaric acids), fumaric acid, maleic acid, succinic acid, chlorogenic acid, salicylic acid, creatine, caffeic acid, bile acids, acetic acid, ascorbic acid, alginic acid, erythorbic acid, polyglutamic acid, glucono-delta-lactone, and their alkali or alkaline earth metal salt derivatives. Additionally, the organic acid additives may be in the D- or L-configuration.

[0128] Suitable organic acid additive salts include, but are not limited to, sodium, calcium, potassium, and magnesium salts of all organic acids, such as citric acid, malic acid, tartaric acid, fumaric acid, lactic acid (e.g., sodium lactate), alginic acid (e.g., sodium alginate), ascorbic acid (e.g., sodium ascorbate), benzoic acid (e.g., sodium benzoate or potassium benzoate), sorbic acid, and adipic acid. Examples of the optionally listed organic acid additives may be substituted with at least one group selected from hydrogen, alkyl, alkenyl, alkynyl, halo, haloalkyl, carboxyl, acyl, acyloxy, amino, amido, carboxyl derivatives, alkylamino, dialkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfo, thiol, imine, sulfonyl, sulfenyl, sulfinyl, sulfamyl, carboxalkoxy, carboxamido, phosphonyl, phosphinyl, phosphoryl, phosphino, thioester, thioether, anhydride, oximino, hydrazino, carbamyl, phosphorus, or phosphonato.

[0129] Suitable inorganic acid additives include, but are not limited to, phosphoric acid, phosphorous acid, polyphosphoric acid, hydrochloric acid, sulfuric acid, carbonic acid, sodium dihydrogen phosphate, and alkali or alkaline earth metal salts thereof (e.g., inositol hexaphosphate Mg / Ca).

[0130] Suitable bitter compound additives include, but are not limited to, caffeine, quinine, urea, orange peel oil, naringin, quascia, and salts thereof.

[0131] Suitable flavoring agents and flavoring ingredient additives include, but are not limited to, vanillin, vanilla extract, mango extract, cinnamon, citrus, coconut, ginger, viridiflorol, almond, menthol (including mint-free menthol), grape skin extract, and grape seed extract. "Flavoring agent" and "flavoring ingredient" are synonymous and may include natural or synthetic substances or combinations thereof. Flavoring agents also include any other substance that imparts flavor and may include natural or non-natural (synthetic) substances that are safe for humans or animals when used within generally acceptable limits. Non-limiting examples of proprietary flavoring agents include Doehler™ Natural Flavoring Sweetness Enhancer K14323 (Doehler™, Darmstadt, Germany), Symrise™ Natural Flavor Mask for Sweeteners 161453 and 164126 (Symrise™, Holzminden, Germany), Natural Advantage™ Bitterness Blockers 1, 2, 9, and 10 (Natural Advantage™, Freehold, New Jersey, USA), and Sucramask™ (Creative Research Management, Stockton, California, USA).

[0132] Suitable polymer additives include, but are not limited to, chitosan, pectin, pectinic acid, pectinic acid, polyuronic acid, polygalacturonic acid, starch, food hydrocolloids or crude extracts thereof (e.g., Acacia Senegal (Fibergum™), Acacia seyal, carrageenan), poly-L-lysine (e.g., poly-L-α-lysine or poly-L-ε-lysine), poly-L-ornithine (e.g., poly-L-α-ornithine or poly-L-ε-ornithine), polypropylene glycol, polyethylene glycol, poly(ethylene glycol methyl ether), polyarginine, polyaspartic acid, polyglutamic acid, polyethyleneimine, alginic acid, sodium alginate, propylene glycol alginate, and sodium polyethyleneglycolalginate, sodium hexametaphosphate and its salts, and other cationic and anionic polymers.

[0133] Suitable protein or protein hydrolysate additives include, but are not limited to, bovine serum albumin (BSA), whey protein (including fractions or concentrates thereof, such as 90% instant whey protein isolate, 34% whey protein, 50% hydrolyzed whey protein, and 80% whey protein concentrate), soluble rice protein, soy protein, protein isolate, protein hydrolysate, reaction products of protein hydrolysates, glycoproteins, and / or proteoglycans containing amino acids (e.g., glycine, alanine, serine, threonine, asparagine, glutamine, arginine, valine, isoleucine, leucine, norvaline, methionine, proline, tyrosine, hydroxyproline, etc.), collagen (e.g., gelatin), partially hydrolyzed collagen (e.g., hydrolyzed fish collagen), and collagen hydrolysates (e.g., porcine collagen hydrolysate).

[0134] Suitable surfactant additives include, but are not limited to, polysorbates (e.g., polyoxyethylene sorbitan monooleate (polysorbate 80), polysorbate 20, polysorbate 60), sodium dodecylbenzenesulfonate, dioctyl sulfosuccinate or dioctyl sodium sulfosuccinate, sodium dodecyl sulfate, cetylpyridinium chloride (hexadecylpyridinium chloride), hexadecyltrimethylammonium bromide, sodium cholate, carbamoyl, choline chloride, sodium glycocholate, sodium taurodeoxycholate, lauric arginate, sodium stearoyl lactylate, sodium taurocholate, lecithin, sucrose oleate, sucrose stearate, sucrose palmitate, sucrose laurate, and other emulsifiers.

[0135] Suitable flavonoid additives are classified as flavonols, flavones, flavanones, flavan-3-ols, isoflavones, or anthocyanidins. Non-limiting examples of flavonoid additives include, but are not limited to, catechins (e.g., green tea extracts such as Polyphenon™ 60, Polyphenon™ 30, and Polyphenon™ 25 (Mitsui Norin Co., Ltd., Japan)), polyphenols, rutin (e.g., enzymatically modified rutin Sammelin™ AO (San-fi Gen FFI, Inc., Osaka, Japan)), neohesperidin, naringin, neohesperidin dihydrochalcone, and the like.

[0136] Suitable alcohol additives include, but are not limited to, ethanol.

[0137] Suitable astringent compound additives include, but are not limited to, tannic acid, europium chloride (EuCl), gadolinium chloride (GdCl), terbium chloride (TbCl), alum, tannic acid, and polyphenols (e.g., tea polyphenols).

[0138] The compositions provided herein may also include one or more functional ingredients, which provide the composition with a proven or recognized health benefit, including, but not limited to, saponins, antioxidants, dietary fiber sources, fatty acids, vitamins, glucosamine, minerals, preservatives, hydrating agents, probiotics, prebiotics, postbiotics, weight management agents, osteoporosis management agents, phytoestrogens, long chain primary aliphatic saturated alcohols, phytosterols, and combinations thereof.

[0139] Exemplary functional ingredients include, but are not limited to, saponins, antioxidants, dietary fiber sources, fatty acids, vitamins, glucosamine, minerals, preservatives, hydration agents, probiotics, prebiotics, weight management agents, osteoporosis management agents, phytoestrogens, long chain primary aliphatic saturated alcohols, phytosterols, and combinations thereof.

[0140] In certain embodiments, the functional ingredient is at least one saponin. As used herein, "at least one saponin" can include a single saponin or multiple saponins as a functional ingredient in the compositions provided herein. Saponins are glycosidic natural plant products containing an aglycone ring structure and one or more sugar moieties. Non-limiting examples of specific saponins for use in certain embodiments of the present invention include Group A acetylsaponins, Group B acetylsaponins, and Group E acetylsaponins. Some common sources of saponins include soybeans, which have a saponin content of approximately 5% by dry weight; soapwort plants (Saponaria), whose roots have historically been used for soap; and alfalfa, aloe, asparagus, grapes, chickpeas, yucca, and various other legumes and weeds. Saponins can be obtained from these sources using extraction techniques well known to those skilled in the art. A description of conventional extraction techniques can be found in US Patent Application Publication No. 2005 / 0123662, the disclosure of which is expressly incorporated by reference.

[0141] In certain embodiments, the functional ingredient is at least one antioxidant. As used herein, "antioxidant" refers to any substance that inhibits, suppresses, or reduces oxidative damage to cells and biomolecules. Examples of suitable antioxidants for embodiments of the present invention include, but are not limited to, vitamins, vitamin cofactors, minerals, hormones, carotenoids, carotenoid terpenoids, non-carotenoid terpenoids, flavonoid polyphenols (e.g., bioflavonoids), flavonols, flavones, phenols, polyphenols, esters of phenols, esters of polyphenols, non-flavonoid phenols, isothiocyanates, and combinations thereof. In some embodiments, the antioxidant is selected from the group consisting of vitamin A, vitamin C, vitamin E, ubiquinone, the mineral selenium, manganese, melatonin, alpha-carotene, beta-carotene, lycopene, lutein, zeanthin, crypoxanthin, reservatol, eugenol, quercetin, catechin, gossypol, hesperetin, curcumin, ferulic acid, thymol, hydroxytyrosol, tumeric, thyme, olive oil, lipoic acid, glutathione, gutamine, oxalic acid, tocopherol-derived compounds, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediaminetetraacetic acid (EDTA), tert-butylhydroxybenzoate (TBHQ), hydroxybenzoates ( ... Non, acetic acid, pectin, tocotrienols, tocopherol, coenzyme Q10, zeaxanthin, astaxanthin, canthaxanthin, saponin, limonoids, kaempfedrol, myricetin, isorhamnetin, proanthocyanidins, quercetin, rutin, luteolin, apigenin, tangeritin, hesperetin, naringenin, erodictyol, flavan-3-ols (e.g., anthocyanidins), gallocatechin, epicatechin, and its gallate ester form, epigallocatechin, and its gallate ester form (ECGC), theaflavin, and its gallate ester form, thearubigins, isoflavones, phytoestrogens, genistein, daidzein, glycitein,Anythocyanins, cyaniding, delphinidin, malvidin, pelargonidin, peonidin, petunidin, ellagic acid, gallic acid, salicylic acid, rosmarinic acid, cinnamic acid and its derivatives (e.g., ferulic acid), chlorogenic acid, chicoric acid, gallotannins, ellagitannins, anthoxanthins, betacyanins and other plant pigments, silymarin, citric acid, lignans, antinutrients, bilirubin, uric acid, R- Lipoic acid, N-acetyl cysteine, emblicanin, apple extract, apple peel extract (Applephenon), red rooibos extract, green rooibos extract, hawthorn berry extract, red raspberry extract, green coffee antioxidant (GCA), aronia extract 20%, grape seed extract (VinOseed), cocoa extract, hops extract, mangosteen extract, mangosteen husk extract, cranberry extract, pomegranate extract, pomegranate husk extract, pomegranate seed extract, hawthorn berry extract, pomegranate extract, cinnamon bark extract, grape skin extract, bilberry extract, pine bark extract, pycnogenol, elderberry extract, morus alba root bark extract, wolfberry (goji) extract, blackberry extract, blueberry extract, blueberry leaf extract, raspberry extract, turmeric extract, citrus bioflavonoids, blackcurrant, ginger, acai powder, green coffee bean extract, green tea extract, and phytic acid, or a combination thereof. In alternate embodiments, the antioxidant is a synthetic antioxidant such as, for example, butylated hydroxytolune or butylated hydroxyanisole. Other sources of suitable antioxidants for embodiments of this invention include, but are not limited to, fruits, vegetables, tea, cocoa, chocolate, spices, herbs, rice, organ meats, yeast, whole grains, or cereals.

[0142] Certain antioxidants belong to a class of phytonutrients called polyphenols (also known as "polyphenolics"), a group of chemicals found in plants characterized by the presence of two or more phenolic groups per molecule. Various health benefits can be derived from polyphenols, including, for example, prevention of cancer, heart disease, and chronic inflammatory diseases, as well as improved mental and physical strength. Suitable polyphenols for embodiments of the present invention include catechins, proanthocyanidins, procyanidins, anthocyanins, quercerin, rutin, reservatrol, isoflavones, curcumin, punicalagin, ellagitannins, hesperidin, naringin, citrus flavonoids, chlorogenic acid, other analogs, and combinations thereof.

[0143] In certain embodiments, the antioxidant is a catechin, such as epigallocatechin gallate (EGCG). In other embodiments, the antioxidant is selected from proanthocyanidins, procyanidins, or combinations thereof. In certain embodiments, the antioxidant is an anthocyanin. In yet other embodiments, the antioxidant is selected from quercetin, rutin, or combinations thereof. In yet other embodiments, the antioxidant is reservatrol. In yet further embodiments, the antioxidant is an isoflavone. In yet further embodiments, the antioxidant is curcumin. In other embodiments, the antioxidant is selected from punicalagins, ellagitannins, or combinations thereof. In yet other embodiments, the antioxidant is chlorogenic acid.

[0144] In certain embodiments, the functional ingredient is at least one source of dietary fiber. Many polymeric carbohydrates, with structures that vary significantly in both composition and linkage, fall within the definition of dietary fiber. Such compounds are well known to those skilled in the art, and non-limiting examples include non-starch polysaccharides, lignin, cellulose, methylcellulose, hemicellulose, β-glucans, pectins, gums, mucilages, waxes, inulin, oligosaccharides, fructooligosaccharides, cyclodextrins, chitin, and combinations thereof. Dietary fiber is generally derived from plant sources, although indigestible animal products, such as chitin, are also classified as dietary fiber. Chitin is a polysaccharide composed of acetylglucosamine units linked by β(1-4) linkages similar to those in cellulose.

[0145] In certain embodiments, the functional ingredient is at least one fatty acid. As used herein, "fatty acid" refers to any straight-chain monocarboxylic acid, including saturated fatty acids, unsaturated fatty acids, long-chain fatty acids, medium-chain fatty acids, short-chain fatty acids, fatty acid precursors (including omega-9 fatty acid precursors), and esterified fatty acids. As used herein, "long-chain polyunsaturated fatty acid" refers to any polyunsaturated carboxylic acid or organic acid with a long aliphatic tail. As used herein, "omega-3 fatty acid" refers to any polyunsaturated fatty acid having its first double bond as the third carbon-carbon bond from the terminal methyl end of its carbon chain. In certain embodiments, omega-3 fatty acids can include long-chain omega-3 fatty acids. As used herein, "omega-6 fatty acid" refers to any polyunsaturated fatty acid having its first double bond as the sixth carbon-carbon bond from the terminal methyl end of its carbon chain.

[0146] Suitable omega-3 fatty acids for use in embodiments of the present invention can be derived from, for example, algae, fish, animals, plants, or combinations thereof.Examples of suitable omega-3 fatty acids include, but are not limited to, linolenic acid, α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, stearidonic acid, eicosatetraenoic acid, and combinations thereof.In some embodiments, suitable omega-3 fatty acids can be provided in fish oil (for example, menhaden oil, tuna oil, salmon oil, bonito oil, and cod oil), microalgae omega-3 oil, or combinations thereof. In certain embodiments, suitable omega-3 fatty acids may be derived from commercially available omega-3 fatty acid oils such as Microalgae DHA oil (from Martek, Columbia, MD), OmegaPure (from Omega Protein, Houston, TX), Marinol C-38 (from Lipid Nutrition, Channahon, IL), Bonito oil and MEG-3 (from Ocean Nutrition, Dartmouth, NS), Evogel (from Symrise, Holzminden, Germany), Marine Oil derived from tuna or salmon (from Arista Wilton, CT), OmegaSource 2000, Marine Oil derived from menhaden, and Marine Oil derived from cod (from OmegaSource, RTP, NC).

[0147] Suitable omega-6 fatty acids include, but are not limited to, linoleic acid, gamma-linolenic acid, dihomo-gamma-linolenic acid, arachidonic acid, eicosadienoic acid, docosadienoic acid, adrenic acid, docosapentaenoic acid, and combinations thereof.

[0148] Suitable esterified fatty acids for embodiments of the present invention may include, but are not limited to, monoacylgycerols containing omega-3 and / or omega-6 fatty acids, diacylgycerols containing omega-3 and / or omega-6 fatty acids, or triacylgycerols containing omega-3 and / or omega-6 fatty acids, and combinations thereof.

[0149] In certain embodiments, the functional ingredient is at least one vitamin. Suitable vitamins include vitamin A, vitamin D, vitamin E, vitamin K, vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin B12, and vitamin C.

[0150] Various other compounds have been classified as vitamins by several authorities. These compounds are sometimes called pseudovitamins and include, but are not limited to, compounds such as ubiquinone (coenzyme Q10), pangamic acid, dimethylglycine, taestrile, amygdalin, flavanoids, para-aminobenzoic acid, adenine, adenylic acid, and s-methylmethionine. As used herein, the term vitamin includes pseudovitamins. In some embodiments, the vitamin is a fat-soluble vitamin selected from vitamins A, D, E, K, and combinations thereof. In other embodiments, the vitamin is a water-soluble vitamin selected from vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B12, folic acid, biotin, pantothenic acid, vitamin C, and combinations thereof.

[0151] In certain embodiments, the functional ingredient is glucosamine, optionally further comprising chondroitin sulfate.

[0152] In certain embodiments, the functional ingredient is at least one mineral. Minerals, according to the teachings of the present invention, contain inorganic chemical elements required by living organisms. Minerals are made up of a wide range of compositions (e.g., elements, simple salts, and complex silicates) and vary widely in crystal structure. They can occur naturally in foods and beverages, be added as dietary supplements, or be ingested or administered separately from foods or beverages.

[0153] Minerals can be classified as either bulk minerals, which are required in relatively large amounts, or trace minerals, which are required in relatively small amounts, with bulk minerals generally required in amounts of about 100 mg or more per day, and trace minerals being those required in amounts less than about 100 mg per day.

[0154] In one embodiment, the minerals are selected from bulk minerals, trace minerals, or combinations thereof. Non-limiting examples of bulk minerals include calcium, chloride, magnesium, phosphorus, potassium, sodium, and sulfur. Non-limiting examples of trace minerals include chromium, cobalt, copper, fluorine, iron, manganese, molybdenum, selenium, zinc, and iodine. Iodine is generally classified as a trace mineral, but is required in greater amounts than other trace minerals and is often classified as a bulk mineral.

[0155] In certain embodiments, the mineral is a trace mineral considered necessary for human nutrition, non-limiting examples of which include bismuth, boron, lithium, nickel, rubidium, silicon, strontium, tellurium, tin, titanium, tungsten, and vanadium.

[0156] The minerals embodied herein may be in any form known to those skilled in the art. For example, in certain embodiments, the minerals may be in their ionic form, having a positive or negative charge. In other particular embodiments, the minerals may be in their molecular form. For example, sulfur and phosphorus often exist naturally as sulfates, sulfides, and phosphates.

[0157] In certain embodiments, the functional ingredient is at least one preservative. In certain embodiments of the present invention, the preservative is selected from antimicrobial agents, antioxidants, antienzymatic agents, or combinations thereof. Non-limiting examples of antimicrobial agents include sulfites, propionates, benzoates, sorbates, nitrates, nitrites, bacteriocins, salts, sugars, acetic acid, dimethyl dicarbonate (DMDC), ethanol, and ozone. In one embodiment, the preservative is a sulfite. Sulfites include, but are not limited to, sulfur dioxide, sodium bisulfite, and potassium bisulfite. In another embodiment, the preservative is a propionate. Propionates include, but are not limited to, propionic acid, calcium propionate, and sodium propionate. In yet another embodiment, the preservative is a benzoate. Benzoates include, but are not limited to, sodium benzoate and benzoic acid. In yet a further embodiment, the preservative is a sorbate. Sorbates include, but are not limited to, potassium sorbate, sodium sorbate, calcium sorbate, and sorbic acid. In yet further embodiments, the preservative is a nitrate and / or nitrite. Nitrates and nitrites include, but are not limited to, sodium nitrate and sodium nitrite. In another embodiment, the at least one preservative is a bacteriocin, such as nisin. In a further embodiment, the preservative is ethanol. In yet another embodiment, the preservative is ozone. Non-limiting examples of antienzyme agents suitable for use as preservatives in certain embodiments of the present invention include ascorbic acid, citric acid, and metal chelators such as ethylenediaminetetraacetic acid (EDTA).

[0158] In certain embodiments, the functional ingredient is at least one hydrating agent. In certain embodiments, the hydrating agent is an electrolyte. Non-limiting examples of electrolytes include sodium, potassium, calcium, magnesium, chloride, phosphate, bicarbonate, and combinations thereof. Electrolytes suitable for use in certain embodiments of the present invention are also described in U.S. Pat. No. 5,681,569, the disclosure of which is expressly incorporated herein by reference. In one embodiment, the electrolyte is derived from its corresponding water-soluble salt. Non-limiting examples of salts for use in certain embodiments include chloride, carbonate, sulfate, acetate, bicarbonate, citrate, phosphate, hydrogen phosphate, tartrate, sorbate, citrate, benzoate, or combinations thereof. In other embodiments, the electrolyte is provided by juice, fruit extract, vegetable extract, tea, or tea extract.

[0159] In certain embodiments of the present invention, the hydrating agent is a carbohydrate that replenishes energy stores burned by muscles. Suitable carbohydrates for use in certain embodiments of the present invention are described in U.S. Patent Nos. 4,312,856, 4,853,237, 5,681,569, and 6,989,171, the disclosures of which are expressly incorporated herein by reference. Non-limiting examples of suitable carbohydrates include monosaccharides, disaccharides, oligosaccharides, complex polysaccharides, or combinations thereof. Non-limiting examples of suitable types of monosaccharides for use in certain embodiments include trioses, tetroses, pentoses, hexoses, heptoses, octose, and nonoses. Non-limiting examples of specific types of suitable monosaccharides include glyceraldehyde, dihydroxyacetone, erythrose, threose, erythrulose, arabinose, lyxose, ribose, xylose, ribulose, xylulose, allose, altrose, galactose, glucose, gulose, idose, mannose, talose, fructose, psicose, sorbose, tagatose, mannoheptulose, sedopertulose, octolose, and sialic acid. Non-limiting examples of suitable disaccharides include sucrose, lactose, and maltose. Non-limiting examples of suitable oligosaccharides include sucrose, maltotriose, and maltodextrin. In other specific embodiments, the carbohydrate is provided by corn syrup, beet sugar, cane sugar, juice, or tea.

[0160] In another specific embodiment, the hydrating agent is a flavanol that provides cellular rehydration. Flavanols are a type of natural substance found in plants, and generally comprise a 2-phenylbenzopyrone molecular skeleton bound to one or more chemical moieties. Non-limiting examples of suitable flavanols for use in certain embodiments of the present invention include catechin, epicatechin, gallocatechin, epigallocatechin, epicatechin gallate, epigallocatechin 3-gallate, theaflavin, theaflavin 3-gallate, theaflavin 3'-gallate, theaflavin 3,3'-gallate, thearubigin, or combinations thereof. Some common sources of flavanols include tea plants, fruits, vegetables, and flowers. In a preferred embodiment, flavanols are extracted from green tea.

[0161] In certain embodiments, the hydration agent is a glycerol solution that enhances exercise endurance. Ingestion of glycerol-containing solutions has been shown to provide beneficial physiological effects, such as expanding blood volume, reducing heart rate, and lowering rectal temperature.

[0162] In certain embodiments, the functional ingredient is selected from at least one probiotic, prebiotic, and combinations thereof. Probiotics are beneficial microorganisms that affect the natural gastrointestinal flora of the human body. Examples of probiotics include, but are not limited to, bacteria from the genera Lactobacilli, Bifidobacteria, Streptococci, or combinations thereof that provide beneficial effects to humans. In certain embodiments of the present invention, at least one probiotic is selected from the genus Lactobacilli. According to other specific embodiments of the present invention, the probiotic is selected from the genus Bifidobacteria. According to yet other specific embodiments of the present invention, the probiotic is selected from the genus Streptococcus.

[0163] Probiotics that can be used in accordance with the present invention are well known to those skilled in the art. Non-limiting examples of foodstuffs containing probiotics include yogurt, sauerkraut, kefir, kimchi, fermented vegetables, and other foodstuffs containing microbial elements that beneficially affect the host animal by improving the intestinal microbalance.

[0164] Prebiotics, according to the teachings of the present invention, include, but are not limited to, mucopolysaccharides, oligosaccharides, polysaccharides, amino acids, vitamins, nutrient precursors, proteins, and combinations thereof. According to certain embodiments of the present invention, the prebiotic is selected from dietary fibers, including, but not limited to, polysaccharides and oligosaccharides. Non-limiting examples of oligosaccharides classified as prebiotics according to certain embodiments of the present invention include fructooligosaccharides, inulin, isomaltooligosaccharides, lactilol, lactosucrose, lactulose, pyrodextrin, soybean oligosaccharides, transgalactooligosaccharides, and xylooligosaccharides. In other embodiments, the prebiotic is an amino acid. While some known prebiotics break down to provide carbohydrates for probiotics, some probiotics also require amino acids for nutrition.

[0165] Prebiotics are found naturally in a variety of foods, including, but not limited to, bananas, berries, asparagus, garlic, wheat, oats, barley (and other whole grains), flaxseed, tomatoes, Jerusalem artichokes, onions and chicory, green vegetables (e.g., dandelion greens, spinach, collard greens, chard, kale, mustard greens, turnip rape), and legumes (e.g., lentils, kidney beans, chickpeas, navy beans, white beans, black beans).

[0166] In certain embodiments, the functional ingredient is at least one weight management agent.

[0167] As used herein, a "weight management agent" includes an appetite suppressant and / or a thermogenic agent. As used herein, the phrases "appetite suppressant," "appetite saturating composition," "satiety agent," and "satiety component" are synonymous. The phrase "appetite suppressant" describes macronutrients, herbal extracts, exogenous hormones, appetite suppressants, appetite reducers, pharmaceuticals, and combinations thereof that, when delivered in an effective amount, suppress, inhibit, reduce, or otherwise reduce a person's appetite. The phrase "thermogenic agent" describes macronutrients, herbal extracts, exogenous hormones, appetite suppressants, appetite reducers, pharmaceuticals, and combinations thereof that, when delivered in an effective amount, activate or otherwise enhance a person's thermogenesis or metabolism.

[0168] Suitable weight management agents include macronutrients selected from the group consisting of proteins, carbohydrates, dietary fats, and combinations thereof. Consumption of proteins, carbohydrates, and dietary fats stimulates the release of peptides that have appetite suppressing effects. For example, consumption of proteins and dietary fats stimulates the release of the gut hormone cholecystokinin (CCK), and consumption of carbohydrates and dietary fats stimulates the release of glucagon-like peptide 1 (GLP-1).

[0169] Suitable macronutrient weight management agents also include carbohydrates. Carbohydrates generally include sugars, starches, cellulose, and gums, which the body converts into glucose for energy. Carbohydrates are often classified into two categories: digestible carbohydrates (e.g., monosaccharides, disaccharides, and starches) and non-digestible carbohydrates (e.g., dietary fiber). Research has shown that non-digestible carbohydrates and complex polymeric carbohydrates with reduced absorption and digestibility in the small intestine stimulate physiological responses that inhibit food intake. Therefore, the carbohydrates embodied herein preferably include non-digestible carbohydrates or carbohydrates with reduced digestibility. Non-limiting examples of such carbohydrates include polydextrose; inulin; polyols derived from monosaccharides, such as erythritol, mannitol, xylitol, and sorbitol; alcohols derived from disaccharides, such as isomalt, lactitol, and maltitol; and hydrogenated starch hydrolysates. Carbohydrates are described in more detail below.

[0170] In another specific embodiment, the weight management agent is dietary fat. Dietary fat is a lipid containing a combination of saturated and unsaturated fatty acids. Polyunsaturated fatty acids have been shown to have a higher satiety power than monounsaturated fatty acids. Therefore, the dietary fat embodied herein preferably contains polyunsaturated fatty acids, non-limiting examples of which include triacylglycerols.

[0171] In certain embodiments, the weight management agent is an herbal extract. Extracts from many types of plants have been identified as having appetite suppressant properties. Non-limiting examples of plants from which extracts have appetite suppressant properties include plants from the genera Hoodia, Trichocaulon, Caralluma, Stapelia, Orbea, Asclepias, and Camellia. Other embodiments include extracts derived from Gymnema Sylvestre, kola nut, Citrus Aurantium, Yerba Mate, Griffonia Simplicifolia, guarana, myrrh, guggul lipids, and blackcurrant seed oil.

[0172] Herbal extracts can be prepared from any type of plant material or plant biomass. Non-limiting examples of plant materials and biomass include stems, roots, leaves, dried powder obtained from the plant material, and sap or dried sap. Herbal extracts are generally prepared by extracting sap from plants and then spray-drying the sap. Alternatively, solvent extraction procedures can be utilized. After the initial extraction, it may be desirable to further fractionate the initial extract (e.g., by column chromatography) to obtain an herbal extract with enhanced activity. Such techniques are well known to those skilled in the art.

[0173] In certain embodiments, the herbal extract is derived from a plant of the genus Hoodia, which includes species such as H. alstonii, H. currorii, H. dregei, H. flava, H. gordonii, H. jutatae, H. mossamedensis, H. officinalis, H. parviflorai, H. pedicellata, H. pilifera, H. ruschii, and H. triebneri. Hoodia plants are fleshy-stemmed succulents native to South Africa. A Hoodia sterol glycoside known as P57 is thought to be responsible for the appetite suppressant effects of Hoodia species. In another particular embodiment, the herbal extract is derived from a plant of the genus Caralluma, which species include C. indica, C. fimbriata, C. attenuate, C. tuberculata, C. edulis, C. adscendens, C. stalagmifera, C. umbellate, C. penicillata, C. russeliana, C. retrospicens, C. arabica, and C. lasiantha. The Carralluma plant belongs to the same subfamily as Hoodia, the Asclepiadaceae.Caralluma is a small, erect, succulent plant native to India that possesses medicinal properties, such as appetite suppression, generally attributed to glycosides belonging to the pregnane group of glycosides, non-limiting examples of which include caratuberside A, caratuberside B, bouceroside I, bouceroside II, bouceroside III, bouceroside IV, bouceroside V, bouceroside VI, bouceroside VII, bouceroside VIII, bouceroside IX, and bouceroside X. In another specific embodiment, the at least one herbal extract is derived from a plant of the genus Trichocaulon. Trichocaulon plants, like Hoodia, are succulents commonly native to South Africa and include the species T. piliferum and T. officinale. In another specific embodiment, the herbal extract is derived from a plant of the genus Stapelia or Orbea, whose species include S. gigantean and O. variegate, respectively. Both Stapelia and Orbea plants belong to the same subfamily as Hoodia, the Asclepiadaceae family. Without wishing to be bound by any theory, compounds exhibiting appetite suppressant activity are believed to be saponins, such as pregnane glycosides, including stavalosides A, B, C, D, E, F, G, H, I, J, and K. In another specific embodiment, the herbal extract is derived from a plant of the genus Asclepias, which also belongs to the Asclepiadaceae family. Non-limiting examples of Asclepias plants include A. incarnate, A. curassayica, A. syriaca, and A. tuberose.Without wishing to be bound by any theory, it is believed that the extract contains steroidal compounds such as pregnane glycosides and pregnane aglycones that have appetite suppressant effects.

[0174] In certain embodiments, the weight control agent is an exogenous hormone having a weight control effect, non-limiting examples of which include CCK, peptide YY, ghrelin, bombesin and gastrin-releasing peptide (GRP), enterostatin, apolipoprotein A-IV, GLP-1, amylin, somastatin, and leptin.

[0175] In another embodiment, the weight control agent is a pharmaceutical agent, non-limiting examples of which include phentenime, diethylpropion, phendimetrazine, sibutramine, rimonabant, oxyntomodulin, floxetine hydrochloride, ephedrine, phenethylamines, or other psychostimulants.

[0176] In certain embodiments, the functional ingredient is at least one osteoporosis treatment agent. In certain embodiments, the osteoporosis treatment agent is at least one calcium source. According to certain embodiments, the calcium source is any compound containing calcium, including salt complexes, solubilized species, and other forms of calcium. Non-limiting examples of calcium sources include amino acid chelated calcium, calcium carbonate, calcium oxide, calcium hydroxide, calcium sulfate, calcium chloride, calcium phosphate, calcium hydrogen phosphate, calcium dihydrogen phosphate, calcium citrate, calcium malate, calcium citrate malate, calcium gluconate, calcium tartrate, calcium lactate, solubilized species thereof, and combinations thereof.

[0177] According to certain embodiments, the agent for treating osteoporosis is a magnesium source. The magnesium source is any compound containing magnesium, including salt complexes, solubilized species, and other forms of magnesium. Non-limiting examples of magnesium sources include magnesium chloride, magnesium citrate, magnesium gluceptate, magnesium gluconate, magnesium lactate, magnesium hydroxide, magnesium picolate, magnesium sulfate, solubilized species thereof, and mixtures thereof. In another specific embodiment, the magnesium source includes amino acid chelated or creatine chelated magnesium.

[0178] In other embodiments, the osteoporosis agent is selected from vitamins D, C, K, precursors thereof, and / or beta-carotene, and combinations thereof.

[0179] Many plants and plant extracts have also been identified as effective in preventing and treating osteoporosis. Non-limiting examples of suitable plants and plant extracts for use as osteoporosis management agents include Taraxacum and Amelanchier species, as disclosed in U.S. Patent Application Publication No. 2005 / 0106215, and Lindera, Artemisia, Acorus, Carthamus, Carum, Cnidium, Curcuma, Cyperus, Juniperus, as disclosed in U.S. Patent Application Publication No. 2005 / 0079232. ), Prunus, Iris, Cichorium, Dodonaea, Epimedium, Erigonoum, Soya, Mentha, Ocimum, thymus, Tanacetum, Plantago, Spearmint, Bixa, Vitis, Rosemarinus, Rhus, and Anethum species.

[0180] In certain embodiments, the functional ingredient is at least one phytoestrogen. Phytoestrogens are compounds typically found in plants that can be delivered to the human body by ingesting plants or plant parts that contain phytoestrogens. As used herein, "phytoestrogen" refers to any substance that, when introduced into the body, produces an estrogenic effect of any degree. For example, phytoestrogens may bind to estrogen receptors in the body and have a small estrogenic effect. Examples of suitable phytoestrogens for embodiments of the present invention include, but are not limited to, isoflavones, stilbenes, lignans, resorcylic acid lactones, coumestans, coumestrol, equol, and combinations thereof. Suitable sources of phytoestrogens include whole grains, cereals, fiber, fruits, vegetables, black cohosh, agave root, blackcurrant, black rhododendron, chasteberry, Viburnum arvense, angelica root, American holly root, false unicorn root, ginseng root, grounzel herb, licorice, liferoot herb, motherwort herb, peony root, raspberry leaves, rose plants, sage leaves, sarsaparilla root, saw palmetto berry, wild yam root, yarrow flowers, beans, soybeans, and soy products (e.g., flavored beans, soybeans, and soy products). miso, soy flour, soy milk, soy nuts, soy protein isolate, tempeh, or tofu), chickpeas, nuts, lentils, seeds, clover, red clover, dandelion greens, dandelion root, fenugreek seeds, green tea, hops, red wine, flaxseed, garlic, onion, linseed, borage, milkweed, caraway, ginseng, vitex, dates, dill, fennel seed, gotu kola, milk thistle, pennyroyal, pomegranate, artemisia, soy flour, tangy flowers, and kudzu root (pueraria lobata), and the like, and combinations thereof.

[0181] Isoflavones belong to a group of phytonutrients called polyphenols. In general, polyphenols (also known as "polyphenolics") are a group of chemicals found in plants that are characterized by the presence of two or more phenolic groups per molecule.

[0182] Suitable phytoestrogenic isoflavones according to embodiments of the present invention include genistein, daidzein, glycitein, biochanin A, formononetin, their respective naturally occurring glycosides and glycoside conjugates, matairesinol, secoisolariciresinol, enterolactone, enterodiol, organized plant proteins, and combinations thereof.

[0183] Suitable sources of isoflavones for embodiments of this invention include, but are not limited to, soybeans, soybean products, beans, alfalfa sprouts, chickpeas, peanuts, and red clover.

[0184] In certain embodiments, the functional ingredient is at least one long-chain primary aliphatic saturated alcohol. Long-chain primary aliphatic saturated alcohols are a diverse group of organic compounds. The term alcohol refers to the fact that these compounds are characterized by a hydroxyl group (-OH) attached to a carbon atom. Non-limiting examples of specific long chain primary aliphatic saturated alcohols for use in certain embodiments of the present invention include 1-octanol of 8 carbon atoms, 1-nonanol of 9 carbon atoms, 1-decanol of 10 carbon atoms, 1-dodecanol of 12 carbon atoms, 1-tetradecanol of 14 carbon atoms, 1-hexadecanol of 16 carbon atoms, 1-octadecanol of 18 carbon atoms, 1-eicosanol of 20 carbon atoms, 1-docosanol of 22 carbon atoms, 1-tetracosanol of 24 carbon atoms, 1-hexacosanol of 26 carbon atoms, 1-heptacosanol of 27 carbon atoms, 1-octanosol of 28 carbon atoms, 1-nonacosanol of 29 carbon atoms, 1-triacontanol of 30 carbon atoms, 1-dotriacontanol of 32 carbon atoms, and 1-tetracontanol of 34 carbon atoms.

[0185] In a particularly desirable embodiment of the present invention, the long-chain primary aliphatic saturated alcohol is polycosanol, a term for a mixture of long-chain primary aliphatic saturated alcohols composed primarily of the 28-carbon 1-octanosol and the 30-carbon 1-triacontanol, along with smaller concentrations of other alcohols such as the 22-carbon 1-docosanol, the 24-carbon 1-tetracosanol, the 26-carbon 1-hexacosanol, the 27-carbon 1-heptacosanol, the 29-carbon 1-nonacosanol, the 32-carbon 1-dotriacontanol, and the 34-carbon 1-tetracontanol.

[0186] In certain embodiments, the functional ingredient is at least one phytosterol, phytostanol, or a combination thereof. As used herein, the phrases "stanol," "plant stanol," and "phytostanol" are synonymous. Plant stanols and stanols naturally occur in small amounts in many fruits, vegetables, nuts, seeds, grains, legumes, vegetable oils, tree bark, and other plant sources. Sterols are a subgroup of steroids that have a hydroxyl group at C-3. Generally, phytosterols, like cholesterol, have a double bond in the steroid nucleus; however, phytosterols may also contain a substituted side chain (R), such as an ethyl or methyl group, or an additional double bond at C-24. The structure of phytosterols is well known to those skilled in the art.

[0187] At least 44 naturally occurring phytosterols have been discovered, generally derived from plants such as corn, soybean, wheat, and wood oils; however, they can also be synthetically produced to form compositions with properties identical to or similar to those of natural phytosterols. According to certain embodiments of the present invention, non-limiting examples of phytosterols known to those of ordinary skill in the art include 4-desmethylsterols (e.g., β-sitosterol, campesterol, stigmasterol, brassicasterol, 22-dehydrobrassicasterol, and Δ5-avenasterol), 4-monomethylsterols, and 4,4-dimethylsterols (triterpene alcohols) (e.g., cycloartenol, 24-methylenecycloartanol, and cyclobuanol).

[0188] As used herein, the phrases "stanol," "plant stanol," and "phytostanol" are synonymous. Phytostanols are saturated sterol alcohols that occur naturally in very small amounts and can also be produced synthetically, such as by hydrogenation of phytosterols. Non-limiting examples of phytostanols, according to certain embodiments of the present invention, include β-sitostanol, campestanol, cycloartanol, and other triterpene alcohols in saturated form.

[0189] Both phytosterols and phytostanols, as used herein, include various isomers such as α and β isomers (e.g., α-sitosterol and β-sitostanol, which constitute among the most effective phytosterols and phytostanols, respectively, for lowering serum cholesterol in mammals).

[0190] The phytosterols and phytostanols of the present invention may be in the form of esters. Suitable methods for deriving esters of phytosterols and phytostanols are well known to those skilled in the art and are disclosed in U.S. Patent Nos. 6,589,588, 6,635,774, 6,800,317, and U.S. Patent Application Publication No. 2003 / 0045473, the disclosures of which are incorporated herein by reference in their entirety. Non-limiting examples of suitable phytosterol and phytostanol esters include sitosterol acetate, sitosterol oleate, stigmasterol oleate, and their corresponding phytostanol esters. The phytosterols and phytostanols of the present invention may also include their derivatives.

[0191] Generally, the amount of functional ingredient in a composition will vary widely depending on the particular composition and the desired functional ingredient. One of ordinary skill in the art will readily ascertain the appropriate amount of functional ingredient for each composition.

[0192] In one embodiment, the method of preparing a composition comprises combining at least one mogroside of the present invention with at least one sweetener and / or additive and / or functional ingredient.

[0193] VI.Consumables The present invention also provides consumable products comprising at least one mogroside of the present invention or compositions comprising at least one mogroside of the present invention. In certain embodiments, the at least one mogroside is isolated and purified.

[0194] The mogrosides of the present invention or compositions comprising same can be mixed with any known edible or oral composition, referred to herein as a "consumable," which, as used herein, means a material that comes into contact with the mouth of a human or animal, including materials that are placed in the mouth and then expelled from the mouth, and materials that are drunk, eaten, swallowed, or otherwise ingested and that are safe for human or animal consumption when used within generally acceptable limits.

[0195] Exemplary consumable products include pharmaceutical compositions, edible gel mixes and compositions, dental compositions, foodstuffs (confectionery, seasonings, chewing gum, cereal compositions, baked goods, dairy products, and tabletop sweetener compositions), beverages and beverage products.

[0196] For example, a beverage is a consumable product. The beverage may be sweetened or unsweetened. The mogrosides of the present invention or compositions comprising same may be added to the beverage or beverage matrix to sweeten the beverage or enhance its existing sweetness or flavor.

[0197] In certain embodiments, the consumable product comprises at least one mogroside of the present invention at a concentration greater than about 1 ppm, such as, for example, from about 1 ppm to about 1,000 ppm, from about 25 ppm to about 1,000 ppm, from about 50 ppm to about 1,000 ppm, from about 75 ppm to about 1,000 ppm, from about 100 ppm to about 1,000 ppm, from about 200 ppm to about 1,000 ppm, from about 300 ppm to about 1,000 ppm, from about 400 ppm to about 1,000 ppm, from about 500 ppm to about 1,000 ppm, or from about 50 ppm to about 600 ppm.

[0198] The consumable may optionally include additives, additional sweeteners, functional ingredients, and combinations thereof, as described above. Any of the additives, additional sweeteners, and functional ingredients described above may be present in the consumable.

[0199] In one embodiment, the composition is a consumable. In one embodiment, the consumable is a beverage or beverage product. The beverage or beverage product comprises at least one mogroside of the present invention or a composition comprising at least one mogroside of the present invention.

[0200] "Beverage product," as used herein, is a ready-to-drink beverage, beverage concentrate, beverage syrup, or powdered beverage. Suitable ready-to-drink beverages include carbonated and non-carbonated beverages. Carbonated beverages include, but are not limited to, frozen carbonated beverages, enhanced sparkling beverages, cola, fruit-flavored sparkling beverages (e.g., lemon-lime, orange, grape, strawberry, and pineapple), ginger ale, soft drinks, and root beer. Non-carbonated beverages include, but are not limited to, fruit juice, fruit-flavored juice, juice drinks, nectar, vegetable juice, vegetable-flavored juice, sports drinks, energy drinks, enhanced water drinks, enhanced water with vitamins, near-water drinks (e.g., water with natural or synthetic flavorings), coconut water, tea-based beverages (e.g., black tea, green tea, rooibos tea, oolong tea), coffee, cocoa drinks, dairy beverages (e.g., milk drinks, dairy coffee, cafe au lait, milk tea, fruit milk drinks), beverages containing grain extracts, drinkable yogurt, protein drinks, and smoothies.

[0201] Beverage concentrates and beverage syrups are prepared with an initial volume of a liquid matrix (e.g., water) and the desired beverage ingredients. A full-strength beverage is then prepared by adding an additional volume of water. Powdered beverages are prepared by dry-mixing all of the beverage ingredients in the absence of a liquid matrix. A full-strength beverage is then prepared by adding the complete volume of water.

[0202] The beverage comprises a matrix, i.e., a base component into which the ingredients, including the compositions of the present invention, are dissolved. In one embodiment, the beverage comprises drinking-quality water as the matrix, such as, for example, deionized water, distilled water, reverse osmosis water, carbon-treated water, purified water, demineralized water, and combinations thereof. Additional suitable matrices include, but are not limited to, phosphoric acid, phosphate buffer, citric acid, citrate buffer, and carbon-treated water.

[0203] The beverage or beverage product may further comprise at least one additional sweetener and / or functional ingredient and / or additive as described herein.

[0204] It is contemplated that the pH of a composition, e.g., a beverage, does not substantially affect or adversely affect the taste of the sweetener. Non-limiting examples of a beverage pH range can be from about 1.8 to about 10. Further examples include a pH range of from about 2 to about 5. In certain embodiments, the beverage pH can be from about 2.5 to about 4.2. One of skill in the art will understand that the pH of a beverage can vary depending on the type of beverage. For example, a dairy beverage can have a pH greater than 4.2.

[0205] The titratable acidity of the beverage can range, for example, from about 0.01 to about 1.0% by weight of the beverage.

[0206] In one embodiment, the sparkling beverage product has an acidity of from about 0.01 to about 1.0% by weight of the beverage, such as, for example, from about 0.05% to about 0.25% by weight of the beverage.

[0207] The carbonation of the sparkling beverage product may be from 0 to about 2% (w / w), for example, from about 0.1 to about 1.0% (w / w) carbon dioxide or its equivalent.

[0208] The beverage may be caffeinated or decaffeinated.

[0209] The temperature of the beverage may range from, for example, about 4°C to about 25°C, for example, from about 4°C to about 100°C.

[0210] The beverage may be a full-calorie beverage having up to about 120 calories per 8 ounce serving.

[0211] The beverage may be a mid-calorie beverage having up to about 60 calories per 8 ounce serving.

[0212] The beverage may be a low-calorie beverage having up to about 40 calories per 8 ounce serving.

[0213] The beverage may be a zero-calorie beverage having less than about 5 calories per 8 ounce serving.

[0214] In certain embodiments, the composition is a cola drink. The cola drink can be a low-calorie, mid-calorie, or zero-calorie drink.

[0215] In particular embodiments, the beverage is a diet beverage. In more particular embodiments, the beverage is a diet carbonated beverage.

[0216] In certain embodiments, the beverage of the present invention is a flavored water beverage.

[0217] The concentration of the mogrosides of the present invention in the beverage may be at, above, or below the threshold sweetness or flavor perception level of the mogrosides of the present invention.

[0218] In one embodiment, the mogrosides of the present invention are present in the beverage at a concentration greater than about 1 ppm, such as, for example, from about 1 ppm to about 1,000 ppm, from about 25 ppm to about 1,000 ppm, from about 50 ppm to about 1,000 ppm, from about 75 ppm to about 1,000 ppm, from about 100 ppm to about 1,000 ppm, from about 200 ppm to about 1,000 ppm, from about 300 ppm to about 1,000 ppm, from about 400 ppm to about 1,000 ppm, or from about 500 ppm to about 1,000 ppm.

[0219] In a more specific embodiment, the mogrosides of the present invention are, for example, about 25 ppm to about 500 ppm, about 25 ppm to about 400 ppm, about 25 ppm to about 300 ppm, about 25 ppm to about 200 ppm, about 25 ppm to about 100 ppm, about 50 ppm to about 600 ppm, about 50 ppm to about 500 ppm, about 50 ppm to about 400 ppm, about 50 ppm to about 300 ppm, about 50 ppm to about 200 ppm, about 50 ppm to about 100 ppm, about 100 ppm to about 600 ppm, about 100 ppm to about 500 ppm, about 100 ppm It is present in the beverage at a concentration of about 25 ppm to about 600 ppm, such as about 25 ppm to about 400 ppm, about 100 ppm to about 300 ppm, about 100 ppm to about 200 ppm, about 200 ppm to about 600 ppm, about 200 ppm to about 500 ppm, about 200 ppm to about 400 ppm, about 200 ppm to about 300 ppm, about 300 ppm to about 600 ppm, about 300 ppm to about 500 ppm, about 300 ppm to about 400 ppm, about 400 ppm to about 600 ppm, about 400 ppm to about 500 ppm, or about 500 ppm to about 600 ppm.

[0220] In one embodiment, the beverage has a sweetness of 3 degrees Brix or greater, e.g., 4 degrees Brix or greater, 5 degrees Brix or greater, 6 degrees Brix or greater, 7 degrees Brix or greater, 8 degrees Brix or greater, 9 degrees Brix or greater, or 10 degrees Brix or greater.

[0221] In another embodiment, the mogrosides of the present invention are present in the beverage at a concentration effective to provide a beverage of 3° Brix or greater, e.g., 4° Brix or greater, 5° Brix or greater, 6° Brix or greater, 7° Brix or greater, 8° Brix or greater, 9° Brix or greater, or 10° Brix or greater.

[0222] In yet another embodiment, the sweetener composition of the present invention is present in the beverage at a concentration effective to provide a sweetener of 3 degrees Brix or greater, e.g., 4 degrees Brix or greater, 5 degrees Brix or greater, 6 degrees Brix or greater, 7 degrees Brix or greater, 8 degrees Brix or greater, 9 degrees Brix or greater, or 10 degrees Brix or greater.

[0223] VII. Method of Use The compounds and compositions of the present invention can be used to sweeten or enhance the flavor or sweetness of consumables. In certain embodiments, the mogrosides of the present invention can be used to sweeten and / or enhance the flavor of consumables. Consumables comprising at least one mogroside of the present invention have taste attributes closer to those of sucrose-sweetened consumables than consumables lacking at least one mogroside of the present invention.

[0224] In one aspect, the invention is a method of preparing a sweetened consumable, the method comprising: (i) providing a consumable; and (ii) adding at least one mogroside of the invention to the consumable to provide the sweetened consumable.

[0225] In certain embodiments, a method of preparing a sweetened consumable includes (i) providing an unsweetened consumable, and (ii) adding at least one mogroside of the present invention to the unsweetened consumable to provide the sweetened consumable.

[0226] In certain embodiments, the present invention is a method for preparing a sweetened beverage, comprising: (i) providing a beverage; and (ii) adding at least one mogroside of the present invention to the beverage to provide a sweetened beverage.

[0227] In certain embodiments, the present invention is a method for preparing a sweetened beverage, comprising: (i) providing an unsweetened beverage; and (ii) adding at least one mogroside of the present invention to the unsweetened beverage to provide a sweetened beverage.

[0228] In the above methods, the mogroside of the present invention can be provided as such, i.e., in the form of a compound, or in the form of a composition. If provided as a composition, the amount of mogroside in the composition is effective to provide a concentration of mogroside that is at, above, or below the flavor or sweetness perception threshold when the composition is added to a consumable (e.g., a beverage). If the mogroside of the present invention is not provided as a composition, it can be added to the consumable at a concentration that is at, above, or below the flavor or sweetness perception threshold.

[0229] In one embodiment, the present invention is a method for enhancing the sweetness of a consumable, comprising: (i) providing a consumable comprising at least one sweetener in a sweetness-imparting amount; and (ii) adding at least one mogroside of the present invention or a composition comprising same to the consumable to provide a consumable having an enhanced sweetness, wherein the mogroside of the present invention is added to the consumable at a concentration below its sweetness perception threshold. In certain embodiments, the mogroside of the present invention is added to the consumable at a concentration below its sweetness perception threshold.

[0230] In certain embodiments, the present invention is a method for enhancing the sweetness of a beverage, comprising: (i) providing a beverage comprising at least one sweetener in a sweetness-imparting amount; and (ii) adding at least one mogroside of the present invention or a composition comprising same to the beverage to provide a sweetness-enhanced beverage, wherein the mogroside is added to the beverage at a concentration below its sweetness perception threshold. In certain embodiments, the mogroside of the present invention is added to the consumable at a concentration below its sweetness perception threshold.

[0231] In another embodiment, the present invention is a method for enhancing the flavor of a consumable, comprising: (i) providing a consumable comprising at least one flavor ingredient; and (ii) adding at least one mogroside of the present invention or a composition comprising same to the consumable to provide a flavor-enhanced consumable, wherein the mogroside of the present invention is added to the consumable at a concentration below its flavor recognition threshold. In certain embodiments, the mogroside of the present invention is added to the consumable at a concentration below its flavor recognition threshold.

[0232] In certain embodiments, a method for enhancing the flavor of a beverage is provided, comprising: (i) providing a beverage comprising at least one flavor component; and (ii) adding at least one mogroside of the present invention or a composition comprising same to the beverage to provide a flavor-enhanced beverage, wherein the mogroside is added to the beverage at a concentration below the flavor recognition threshold of the mogroside. In certain embodiments, the mogroside of the present invention is added to the consumable at a concentration below its flavor recognition threshold.

[0233] In one embodiment, a method for enhancing the sweetness of a consumable comprises (i) providing a consumable comprising at least one sweetener, and (ii) adding at least one mogroside of the present invention to the composition to provide a sweetness-enhanced composition.

[0234] In another embodiment, a method for enhancing the sweetness of a consumable comprises (i) providing a consumable matrix, and (ii) adding at least one sweetener and at least one mogroside of the present invention to the consumable matrix to provide a sweetness-enhanced consumable. The at least one sweetener and the at least one mogroside of the present invention may be added together, i.e., in the form of a composition, or may be added separately.

[0235] As used herein, the term "consumable matrix" refers to a composition that includes all typical ingredients except a sweetener or sweetener composition.

[0236] In certain embodiments, the SE of a consumable comprising at least one mogroside of the present invention and at least one sweetener is enhanced by at least about 1.2-fold, such as at least about 1.3-fold, at least about 1.4-fold, at least about 1.5-fold, at least about 1.6-fold, at least about 1.7-fold, at least about 1.8-fold, at least about 1.9-fold, and at least about 2.0-fold, compared to the SE of the consumable without the at least one mogroside of the present invention.

[0237] In another embodiment, the addition of at least one mogroside of the present invention to a consumable or consumable matrix increases the Brix by at least 1 degree Brix, e.g., at least 2 degrees Brix, at least 3 degrees Brix, or at least 4 degrees Brix, compared to the Brix of the consumable without the mogroside of the present invention.

[0238] In another aspect, a method for making a consumable taste more closely resemble a sucrose-sweetened consumable comprises: (i) providing a consumable comprising at least one sweetener in a sweetening amount; and (ii) adding at least one mogroside of the present invention in an amount effective to adjust one or more taste attributes of the sweetener, thereby making the consumable taste more closely resemble a sucrose-sweetened consumable compared to the consumable in the absence of the at least one mogroside of the present invention.

[0239] In another embodiment, a method for making a consumable taste more similar to a sucrose-sweetened consumable comprises: (i) providing a consumable matrix; and (ii) adding a sweetening amount of at least one sweetener and at least one mogroside of the present invention to the consumable matrix to provide a consumable that tastes more similar to a sucrose-sweetened consumable, wherein the at least one mogroside of the present invention is present in an amount effective to modulate one or more taste attributes of the sweetener to make the consumable taste more similar to a sucrose-sweetened consumable compared to the consumable in the absence of the at least one mogroside of the present invention. The at least one sweetener and the at least one mogroside of the present invention may be added together, i.e., in the form of a composition, or may be added separately.

[0240] Methods or preparing the sweetness-enhanced compositions are also provided.

[0241] In one embodiment, a method of preparing a composition includes (i) providing a composition comprising at least one sweetener, and (ii) adding at least one mogroside of the present invention to the composition to provide the composition.

[0242] In one aspect, the invention is a method for preparing a sweetened consumable, the method comprising: (i) providing a consumable; and (ii) adding at least one mogroside of the invention to the consumable in a sweetening amount to provide the sweetened consumable.

[0243] In certain embodiments, a method of preparing a sweetened consumable comprises (i) providing an unsweetened consumable, and (ii) adding at least one mogroside of the present invention to the unsweetened consumable in a sweetened amount to provide the sweetened consumable.

[0244] In certain embodiments, the present invention is a method for preparing a sweetened beverage, the method comprising: (i) providing a beverage; and (ii) adding at least one mogroside of the present invention to the beverage in a sweetening amount to provide the sweetened beverage.

[0245] In certain embodiments, the present invention is a method for preparing a sweetened beverage, comprising: (i) providing an unsweetened beverage; and (ii) adding at least one mogroside of the present invention to the unsweetened beverage in a sweetening amount to provide the sweetened beverage.

[0246] VIII. Purification Methods The present invention also extends to a method for purifying the mogrosides of the present invention.

[0247] In one embodiment, the present invention is a method for purifying a mogroside of the invention, the method comprising: (i) passing a solution comprising a raw material comprising a mogroside of the invention through an HPLC column; and (ii) eluting fractions comprising a mogroside of the invention to provide a purified mogroside composition comprising a mogroside of the invention. The HPLC column may be any suitable HPLC preparative or semi-preparative scale column.

[0248] As used herein, the term "preparative HPLC" refers to an HPLC system capable of producing high (500 or more) microgram, milligram, or gram sized product fractions. The term "preparative" includes both preparative and semi-preparative columns, but does not include analytical columns that provide fractions in the nanogram to low microgram range.

[0249] As used herein, an "HPLC-compatible detector" is a detector suitable for use in an HPLC system that is capable of providing a detectable signal upon elution of a compound peak. For example, a detector capable of producing a signal as a compound elutes from a compound is an HPLC-compatible detector. When component absorbances vary widely, it may be necessary to utilize more than one detector. A detector capable of detecting the desired component is not a "non-compatible" detector because it is incapable of detecting undesired peaks.

[0250] An HPLC apparatus typically includes at least the following elements: a column packed with a suitable stationary phase, a mobile phase, a pump for forcing the mobile phase through the column under pressure, and a detector for detecting the presence of compounds eluting from the column. The apparatus may optionally include a means for providing gradient elution, although such is not necessary using the methods described herein. Routine methods and apparatus for performing HPLC separations are well known in the art.

[0251] A suitable stationary phase is one that elutes the compound of interest. Preferred columns can be, but are not limited to, normal-phase columns (neutral, acidic, or basic), reverse-phase columns (of any alkyl chain length), synthetic cross-linked polymer columns (e.g., styrene and divinylbenzene), size-exclusion columns, ion-exchange columns, bioaffinity columns, and any combination thereof. The particle size of the stationary phase is in the range of several microns to several hundred microns.

[0252] Suitable detection devices include, but are not limited to, mass spectrometers, UV detectors, IR detectors, and light scattering detectors. The methods described herein use any combination of these detectors. The most preferred embodiment uses a mass spectrometer and a UV detector.

[0253] As used herein, "raw material" refers to the material being purified by the present method. The raw material contains the mogrosides of the present invention at a purity lower than that provided by the present purification method. The raw material can be a liquid or a solid. Exemplary raw materials include, but are not limited to, mixtures of mogrosides and luo han guo extracts (commercially available or prepared).

[0254] As will be appreciated by those skilled in the art, any solid source material must be brought into solution before performing the HPLC method.

[0255] In one embodiment, the representative analytical HPLC protocol correlates with the preparative or semi-preparative HPLC protocol used to purify the compound.

[0256] In another embodiment, suitable conditions for purifying the mogrosides of the invention can be performed by route scouting representative samples for a given analytical HPLC column, solvent system, and flow rate. In yet another embodiment, correlated preparative or semi-preparative HPLC methods can be applied to purify the mogrosides of the invention with or without the need to modify the purification parameters.

[0257] In some embodiments, the eluent (mobile phase) is selected from the group consisting of water, acetonitrile, methanol, 2-propanol, ethyl acetate, dimethylformamide, dimethyl sulfide, pyridine, triethylamine, formic acid, trifluoroacetic acid, acetic acid, an aqueous solution containing ammonium acetate, heptafluorobutyric acid, and any combination thereof.

[0258] In one embodiment, the HPLC method is isocratic. In another embodiment, the HPLC method is gradient. In yet another embodiment, the HPLC method is stepwise.

[0259] In one embodiment, the impurities are eluted from the HPLC column after the elution of one or more fractions comprising the mogrosides of the invention, hi another embodiment, the impurities are eluted from the HPLC column before the elution of one or more fractions comprising the mogrosides of the invention.

[0260] The method may further comprise removing the solvent from the eluted solution, i.e., drying. In one embodiment, the method further comprises partially removing the solvent from the eluted solution to provide a concentrate comprising the mogrosides of the present invention. In another embodiment, the method further comprises removing substantially all of the solvent from the eluted solution to provide a substantially dry composition comprising the mogrosides of the present invention.

[0261] Removal of the solvent can be accomplished by any means known to those skilled in the art, including, but not limited to, evaporation, distillation, freeze drying, vacuum drying, and spray drying.

[0262] The resulting purified fraction containing the mogrosides of the present invention can be further purified by other methods to increase purity. Suitable methods include, but are not limited to, crystallization, chromatography, extraction, and distillation. Such methods are well known to those skilled in the art.

[0263] The raw material can be a fraction or fractions containing mogrosides of the invention recovered from at least one prior method or HPLC protocol. In one embodiment, multiple fractions from the same prior method or HPLC protocol are pooled, and optionally the solvent is removed, before subjecting the raw material to another method again. In another embodiment, fractions from different prior methods or HPLC protocols are pooled, and optionally the solvent is removed, before subjecting the raw material to another method again.

[0264] In one embodiment, the raw material to be resubjected to an additional process comprises a liquid fraction from one or more earlier (and optionally different) processes mixed with a substantially dry material obtained by drying a fraction from one or more earlier (and optionally different) processes. In another embodiment, the raw material to be resubjected to an additional process comprises a substantially dry material obtained by drying a fraction from one or more earlier (and optionally different) processes, wherein the raw material is brought into solution before the solution is passed through a subsequent HPLC column.

[0265] The second and subsequent methods may have different HPLC protocols (e.g., solvent systems, columns, methods) and different post-elution steps (e.g., partial removal of solvent, complete removal of solvent, elution of impurities, use of crystallization or extraction).

[0266] The isolated material can be subjected to further processing two, three, four or more times, each time providing greater purity of the purified mogrosides of the invention.

[0267] In one embodiment, the method provides a purified mogroside composition comprising a mogroside of the invention that is at least about 80% by weight or greater in purity, e.g., at least about 85% by weight, at least about 90% by weight, at least about 95% by weight, or at least about 97% by weight or greater. In another embodiment, the purification provides a pure mogroside of the invention, i.e., greater than 99% by weight on a dry basis.

[0268] The advantages of the present invention will become more apparent from the detailed description given below. It should be understood, however, that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of example only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from the detailed description. [Example]

[0269] Example Example 1: Mogroside V Bioconversion LC-MS was performed on MV90 (90% mogroside V) substrate (Hunan Huacheng Biotech Inc.). Two prominent peaks are observed in the LC-MS extracted for the masses of xylose-containing mogrosides with and without sodium 5-glycosylation ("xylose-containing mogroside 1" and "xylose-containing mogroside 2"). The results are shown in Figure 1.

[0270] Using DSM Maxilact enzymes in bioconversion reactions, the following reactions are expected to occur: [ka]

[0271] In one embodiment, a reaction is provided in which an isomogroside V-like xylose-containing compound is converted to an isomogroside IV-like xylose-containing compound. [ka]

[0272] One or both of these can be converted to xylose-containing mogrosides with four glycosylations.

[0273] Based on knowledge of the structures of mogrosides containing 5-glucosylation (i.e., glucose-only mogrosides), xylose-containing mogrosides with five possible glycosyl units resemble mogroside V and isomogroside V.

[0274] Example 2: Mogroside diversification To prepare mogroside compounds using the mogroside V bioconversion pathway, various enzymes were screened for use in the addition of non-glucose monosaccharides to the C24 glycosyl group of MogIIIE. [ka]

[0275] Materials and equipment: Glycotransferases and transglycosylating glycosidases were selected that are capable of adding α / β non-glucose sugars to glucose positions (not only C6, but also C3 and C4). Enzymes useful for the inversion reaction include, but are not limited to, α-rhamnosidase 78A (Megazyme, E-RHAMS), β-fructofuranosidase (invertase) (Megazyme, E-INVRT), α-galactosidase (Megazyme, E-AGALPS), and α-xylosidase (Megazyme, E-AXSEC).

[0276] Reaction mixture: 50 μL reaction (40 hours at 40°C): Enzyme 1 μL 1M sugar 4μL 100mM MogIIIE 10μL 2x buffer, various pH values, 25 μL 10 μL H2O

[0277] Set up a pH ladder to assess changes in bioconversion yield: Acetate buffer 3.6; Acetate buffer 4.6; Acetate buffer 5.6; phosphate buffer 6.6; Phosphate buffer solution 7.0 The inventors applied the above two tables to prepare buffer solutions with different pH values (4.6, 5.6, 6.6).

[0278] Refining Process The reaction mixture was purified by C18 column on an HPLC system.

[0279] 1. Rhamnose 25mL reaction (60 hours at 40°C): α-rhamnosidase 0.5mL 12.5 mL of 500 mM rhamnose in 2x buffer, pH 5.6 130mM MogIIIE 11mL At 30 hours, another 0.5 mL α-rhamnosidase was added. After 60 hours, the reaction was boiled, filtered and subjected to HPLC purification, which is shown in Figure 2. After C18 HPLC purification (10–40% acetonitrile), approximately 24 mg of peak 45 was recovered, and peak 46 consisted of two inseparable compounds. 47 + 48 were the substrate MogIIIE. Peak 45 was subjected to NMR analysis and determined to be CC-00500. The reaction at pH 4.6 to 5.6 had relatively high productivity. The isolated HPLC peaks were subjected to mass spectrometry analysis, shown in Figure 3. Both isolated HPLC peaks showed an increase in mass from 985 to 1131, as expected for the addition of 1 rhamnose to the mogroside IIIE substrate.

[0280] 2. Galactose 25mL reaction (60 hours at 40°C): α-galactosidase 0.5mL 12.5 mL of 500 mM rhamnose in 2x buffer, pH 4.6 130mM MogIIIE 11mL At 30 hours, another 0.5 mL α-galactosidase was added. After 60 hours, the reaction was boiled, filtered, and then subjected to HPLC purification, which is shown in Figure 4. After C18 HPLC purification (10-40% acetonitrile), approximately 4 mg of peak 108 was recovered, and approximately 28 mg of peak 109 was recovered. Peak 110 contained a large amount of substrate MogIIIE. Peaks 111 and 112 were substrate MogIIIE. Peaks 108 and 109 were subjected to NMR analysis, and peak 108 was identified as CC-00489. Reactions at pH 3.6-4.6 had relatively high productivity. The isolated HPLC peaks were subjected to mass spectrometry analysis, shown in Figure 5. Both isolated HPLC peaks showed an increase in mass from 985 to 1147, as expected for the addition of 1 rhamnose to the mogroside IIIE substrate.

[0281] 3. Xylose 25mL reaction (60 hours at 40°C): α-galactosidase 0.5mL 12.5 mL of 500 mM xylose in 2x buffer, pH 4.6 130mM MogIIIE 11mL At 30 hours, another 0.5 mL α-galactosidase was added. At 60 hours, the reaction was boiled, filtered, and then subjected to HPLC purification. The reaction at pH 4.6 had a relatively high productivity, which is shown in Figure 6. After C18 HPLC purification (10–40% acetonitrile), peaks 36, 37, 38, 39, and 40 were collected. Mass spectrometry analysis of peak 39 showed a small peak with m / z of 1117, suggesting the addition of xylose to mogroside IIIE, as shown in Figure 7 .

[0282] Conclusion: MALDI-TOF analysis revealed that the productivity of adding galactose, rhamnose, or xylose was relatively high.

[0283] Example 3: Evaluation of sweetness of CC-00489 The sweetness of mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-xylopyranosyl-(1→6)]-[β-D-glucopyranosyl-(1→2)]-β-D-glucopyranoside} (CC-00489) was determined relative to a sucrose standard.

[0284] Detailed Description: 1. Standards: 8%, 9%, and 10% sucrose in deionized water. 2. The sweetness of the samples was measured relative to a sucrose standard. 3. Test method: Sip and spit 4. Other temporal profiles: bitterness, astringency, lingering sweetness, bitter aftertaste 5. The test level for CC-00489 was 400 ppm. 6. The batch number for CC-00489 was IN-SDV-D-196-2.

[0285] Sample preparation

[0286] [Table 1]

[0287] [Table 2]

[0288] The components were added to deionized water with stirring until the solids were visibly dissolved, and the samples were poured into glass bottles or vials and stored at 4°C.

[0289] Taste evaluation The taste test was conducted by two panelists. The bottles / vials were removed from the refrigerator. Approximately 25 ml of the sucrose solution was poured into a 4 oz. plastic cup, and 3 ml of the sweetener solution was poured into a 2 oz. plastic cup. The panelists were given mineral water to rinse their mouths before tasting and between tasting different samples. The panelists were also given a non-salty cracker to eat, followed by mineral water to rinse their mouths before tasting the next sample.

[0290] First, panelists were asked to identify the sweetness of three sucrose solutions in deionized water (Table 2).

[0291] Panelists were given samples of Compound A in deionized water (Table 1). Panelists were asked to measure the sweetness of the sample relative to a sucrose standard (Table 2) and describe the taste profile, including bitterness, astringency, and a bitter aftertaste. Panelists were instructed to take a sip, evaluate the sweetness, and then spit the sample into a cup provided for that purpose.

[0292] One panelist rated the sweetness of the Compound A sample as being 9-10 sucrose equivalents (SE) with a mild astringency and a good texture, while another panelist rated the sweetness of the sample as being close to 10 SE.

[0293] Example 4: Purification with C18 resin Mogrosides of interest (e.g., CC-00520) were isolated following bioconversion of MV90 (90% mogroside V) substrate (Hunan Huacheng Biotech Inc.) to siamenoside I using a β-galactosidase enzyme (e.g., Aspergillus oryzae β-galactosidase (AoBG) such as Maxilact A4).

[0294] 340 g of MV90 was dissolved in 1 L of potassium phosphate buffer (100 mM, pH 6.5) and diluted with 1 L of water (total of 2 L for a final buffer concentration of 50 mM), which was then filtered through a 0.2 μm sterile filter unit.

[0295] Six liters of recently concentrated Maxilact A4 (3.5x concentrated) was diluted with 6 liters of buffer (100 mM potassium phosphate, pH 6.5), and the pH of the diluted enzyme was adjusted to approximately 6.0. 1% filter aid (Celite; w / v) was added, and the suspension was mixed for approximately 15 minutes. The suspension was then filtered through a tub equipped with a coarse frit. The filtrate was passed through a series of capsule filters (5 μm and 0.2 μm) before a final filtration into a sterile filtration unit. The final volume of filtered enzyme was approximately 11 L.

[0296] 11 L of sterile filtered enzyme and 2 L of sterilized Mog.V solution were transferred to fermenters separately. The pH was adjusted to 6.2, and the reaction temperature was adjusted to 50°C to start the reaction.

[0297] After the bioconversion reaction, the mogrosides of the present invention were separated from the enzymes and salts. To separate the protein from the mogrosides, the reaction mixture was mixed with sodium hydroxide to raise the pH to 12.4. Ethanol was added to create a 20% ethanolic solution. The mixture was filtered through a 10 kDa Koch Romicon membrane at an inlet pressure of 1.7 bar and atmospheric pressure at the outlet. The pH of the permeate was lowered to 5.5 using acetic acid and allowed to cool overnight. The next day, the solution was refiltered through a 10 kDa Koch Romicon membrane.

[0298] Water, ethanol, and salts were removed using a Koch SR3D nanofiltration membrane with a 200 Da cutoff. The solution was diafiltered until the ethanol concentration was less than 3 percent and concentrated to 20-30 L. The concentrated mogrosides were mixed with a water / ammonia acetate solution to bring the solution up to approximately 110 L.

[0299] The mixture was passed through a Biotage SNAP KP-C18-HS 400 g guard cartridge. The resulting solution was mixed and dispensed into 5 L HDPE jerry cans.

[0300] Chromatography There are six steps in a chromatographic separation: 1) Column equilibration in preparation for loading. 2) Column loading: After packing, a small amount of equilibration solution distributes the mogrosides throughout the bed. 3) Removal of mogroside V and other early eluting compounds. The first approximately 120 L was sampled and sent to waste. The next approximately 27 kg was collected, with the target purity being present in the last fraction. 4) Removal of siamenoside I, two large 18 kg fractions followed by 4 x 4.5 kg fractions. The last fraction did not meet purity specifications. 5) Straight XNS grade ethanol (95%). The first 18 kg for recovery was the Mog IIIe fraction. 6) Straight XNS-grade ethanol for washing the column.

[0301] The compositions of the eluents are shown in Table 14.

[0302] [Table 3]

[0303] diafiltration The fractions containing the mogrosides of the present invention were further processed. The fractions containing the mogrosides of the present invention were identified, for example, by HPLC-MS techniques and typically co-eluted with siamenoside I. Ethanol and ammonia acetate were removed. The combined fractions were diafiltered and RO water was added to the solution to maintain the ethanol concentration below 15%. Ammonia, acetate, ethanol, and water were removed.

[0304] A secondary HPLC chromatography was performed, for example on another C18 column, to further purify the mogrosides of the invention. Fractions were lyophilized for analysis and storage.

[0305] Example 5: Synthesis of CC-00491 3-O-β-D-Glucopyranosylmogrol 24-O-β-D-glucopyranosyl-(1→2)-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (Scheme 1). To a solution of 6 (562.5 mg, 0.24 mmol) in dry THF (10 mL) and dry MeOH (10 mL) was added dropwise a 0.5 M solution of NaOMe in MeOH (0.24 mL, 0.12 mmol) at room temperature under a N atmosphere. The mixture was stirred at the same temperature for 18 h, neutralized with 10% aqueous AcOH, and concentrated under reduced pressure. The remaining white solid was purified by preparative HPLC (Phenomenex Luna C18 column, 250 × 30 mm, 10 μm particles, HO → CH3CN gradient, 40 mL / min, t RThe product was purified by HPLC (HPLC = 12.5 min). Fractions with retention times between 11 and 15 min were collected and combined based on HPLC and LCMS analysis. The product was redissolved in water and dried in a lyophilizer to give 7 (172 mg, 64%) as a white powder. Melting point = 179-182 °C. f = 0.56 (silica gel, CH2Cl2 / MeOH / H2O = 5:4:1). 1 H NMR(500MHz,pyridine-d5)δ 7.53(d,J=5.0Hz,1H),7.51(d,J=3.0Hz,1H),7.44(d,J=3.0Hz,1H),7.30(d,J=3.8Hz,1H),7.19-7.12(m,2H),7. 11-6.99(m,2H),6.65-6.53(m,2H),6.46(d,J=6.2Hz,1H),6.22-6.13(m,1H),5.66(d,J=5.8Hz,1H),5.47(d,J=6 .4Hz,1H),5.46(d,J=1.1Hz,1H),5.43(t,J=5.9Hz,1H),5.36(d,J=7.8Hz,1H),5.16(s,1H),5.04(d,J=7.8Hz,1H ),4.91(d,J=7.8Hz,1H),4.70-4.65(m,1H),4.64(d,J=9.7Hz,1H),4.60-4.48(m,3H),4.46-4.32(m,3H),4.31-4 .21(m,5H),4.20-4.01(m,6H),4.01-3.92(m,4H),3.92-3.86(m,1H),3.68(s,1H),3.02-2.88(m,1H),2.86-2.72 (m,1H),2.52-2.39(m,1H),2.36-2.22(m,1H),2.20-1.94(m,6H),1.90-1.78(m,3H),1.78-1.70(m,1H),1.69-1. 60(m,2H),1.65(d,J=6.2Hz,3H),1.59-1.48(m,1H),1.57(s,3H),1.52(s,3H),1.48-1.38(m,1H),1.44(s,3H),1 .33(s,3H),1.20-1.11(m,1H),1.14(s,3H),1.10-1.00(m,1H),1.08(d,J=6.4Hz,3H),0.90(s,3H),0.89(s,3H). 13C NMR (125 MHz, pyridine-d5) δ 144.5, 118.8, 107.8, 106.6, 103.1, 102.7, 89.2, 88.3, 84.1, 79.1, 78.7, 78.7, 78.5, 78.2, 76.9, 76.5, 75.9, 74.5, 73.1, 72.7, 72.7, 72.5, 72.1, 71.8, 70.1, 68.6, 63.8, 63.4 ,51.2,47.8,43.9,42.7,41.5,40.5,37.2,37.1,34.9,34.1,29.9,29.3,28.9,28.0,27.3,27.2,26.7,26.7,26.3,24.9,19.7,19.5,19.1,17.4.ESI-MS(negative mode) m / z:[MH] - 1108. [ka]

[0306] Example 6: Purification and characterization of CC-00507 Materials: The material used for the isolation of CC-00507 (Lot No. IN-RAS-A-53-3) was Luo Han Guo extract (Batch No. LHGE-180125) purchased from Huacheng Bio.

[0307] HPLC analysis: HPLC analysis was performed on an Agilent 1200 system coupled with a variable wavelength detector (VWD) detector. Samples obtained from processing and final purity assessment were run using the method conditions described in Table 1.

[0308] [Table 4]

[0309] Preparative HPLC analysis: Preparative HPLC analysis was performed on an Agilent preparative HPLC coupled with a UV-Vis detector.

[0310] Primary Processing: Approximately 109 g of Luo Han Guo extract (Batch No. LHGE-180125) was processed using the preparative HPLC method described in Table 2. The target fraction retention time was 20.00-23.00 min (Peak ID: LHGE-180125-P4). This material was pooled and lyophilized. The final yield of Peak ID: LHGE-180125-P4 (Lot No. IN-RAS-A-24-4) was approximately 12.2 g.

[0311] [Table 5]

[0312] Secondary Processing: Approximately 12.2 g of #LHGE-180125-P4 was processed using the preparative HPLC method described in Table 3. The target fraction had a retention time of 8.60-9.50 min (Peak ID: LHGE-180125-P4-D). This material was pooled and lyophilized. The final yield of Peak ID: LHGE-180125-P4-D (Lot No. IN-VVP-K-194-4) was approximately 60 mg.

[0313] [Table 6]

[0314] Tertiary Processing: Approximately 60 mg of #LHGE-180125-P4-D was processed using the preparative HPLC method described in Table 4. The target fraction had a retention time of 7.20-7.60 min (Peak ID: LHGE-180125-P4-D3). The material was pooled and lyophilized for isolation. The final yield of Peak ID: LHGE-180125-P4-D3 (Lot No. IN-RAS-A-53-3) was 4.4 mg with a purity of 90.6% (area %).

[0315] [Table 7]

[0316] MS and MS / MS. MS and MS / MS data were generated on a Waters QTof Micro mass spectrometer equipped with an electrospray ionization source. Samples were analyzed by negative ESI. Samples (approximately 0.2 mg) were diluted with 50:50 ACN:HO to a concentration of approximately 0.2 mg / mL and introduced by direct infusion.

[0317] Mass spectrometry. The ESI-TOF mass spectrum obtained by injecting a sample of CC-00507 showed [M-H] at m / z 1255.6326. - The ion [MH] was shown. - The mass of the ion is calculated based on the predicted molecular formula C 59 H 100 O 28 was in good agreement with (C 59 H 99 O 28 Calculated value: 1255.6323, error: 0.2 ppm). MS data confirmed that CC-00507 has the molecular formula C 59 H 100 O 28 The ion observed at m / z 1353.6130 was probably [M-H+H3PO4] - This is due to the following.

[0318] The MS / MS spectrum of CC-00507 shows fragmentation of [M-H] at m / z 1255.5. - Ions were selected to show the loss of a xylose unit at m / z 1123.5382, followed by the sequential loss of four glucose units at m / z 961.4853, 799.4445, 637.3991, and 475.3554. After the loss of the xylose unit from the structure, an alternative fragmentation pathway was also observed in the spectrum, which would correspond to the loss of a water molecule from the central triterpene core, followed by the sequential loss of sugar units at m / z 1105.5109, 943.4888, 781.4421, and 619.3909.

[0319] NMR. Samples were prepared by dissolving the available material in 130 μL of CD3OD, and NMR data were obtained. 1 H, 1 H- 1 H COSY, 1 H- 13 C HSQC-DEPT and 1D TOCSY NMR data were obtained on a Bruker Avance 500 MHz NMR instrument equipped with a 2.5 mm inverse probe. 13 C. 1 H- 13 C HMBC, and 1 H- 1 Additional NMR data, such as H ROESY, was obtained at Rensselaer Polytechnic Institute using their Bruker Avance 600 MHz instrument with a 5 mm cryoprobe. 1 The H NMR spectrum shows δ H Based on the CHD2OD resonance of 3.30, 13 The C NMR spectrum shows δ C The CD3OD resonance at 49.0 was used as the reference.

[0320] [Table 8]

[0321] [Table 9]

[0322] [Table 10]

[0323] Spectroscopic (NMR) and spectrophotometric (MS) analyses of CC-00507 allowed the complete assignment of its structure, which was identified as mogrol-3-O-[{β-D-glucopyranosyl-(1→6)}-β-D-glucopyranoside]-24-O-[{β-D-xylopyranosyl-(1→2)}-{β-D-glucopyranosyl-(1→6)}-β-D-glucopyranoside]. This compound has four glucose units and one xylose unit. The xylose unit is linked to G1 by forming a 1→2 sugar bond. cI is bonded to.

[0324] Example 7: Preparation and characterization of CC-00518 CC-00518 was synthesized by α-D-xylosyl fluoride and α-xylosidase (Megazyme, E-AXSEC) to synthesize mogroside III. E CC-00518 was isolated from the reaction product of the bioconversion of 10 mM α-xylosyl fluoride, 10 mM MogIIIE, 0.01% BSA, 6 units α-xylosidase, in 50 mM phosphate buffer (pH 7.0) at 37°C for approximately 30 minutes. CC-00518 was purified using preparative HPLC in multiple replicates to give approximately 25 mg with 99% purity (area %). A series of 1D and 2D experiments ( 1 H, 13 C. 1 H- 1 H COSY, 1 H- 13 C HSQC-DEPT, 1 H- 13 C HSQC-TOCSY, and 1 H- 13The structure was elucidated by C HMBC as mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]. Spectroscopic data indicated the presence of a central triterpene core and four sugar units: three glucose and one xylose in the structure. The linkages between the sugar units and their attachment to the central triterpene core were determined based on COSY, TOCSY, and HSQC-DEPT data. The glucose unit exists as the β anomer, while the xylose unit ( 3 J = 3.7 Hz) exists in the α configuration.

[0325] [Table 11]

[0326] [Table 12]

[0327] Example 8: Preparation, purification, and characterization of CC-00520 CC-00520 was isolated as the lesser mogroside obtained from the conversion of mogroside V to siamenoside I, e.g., Example 4. The compound is also present in and can be isolated from monk fruit extracts.

[0328] Materials: The material used in the isolation of Lot No. IN-VVP-K-172 (CC-00520) was an R11 purified sample, Lot No. AMR100489-23-F2.

[0329] The material used to isolate Lot No. IN-RAS-A-119-3 (CC-00520) was Monk Fruit Extract Sample, Lot No. LHGE-180125.

[0330] HPLC Analysis: HPLC analysis was performed on an Agilent 1200 system coupled with a variable wavelength (VWD) detector. Samples from multiple processes and final purity assessments were run using the method conditions listed in Tables 1 and 2.

[0331] [Table 13]

[0332] [Table 14]

[0333] Primary Processing: Approximately 68 g of Lot No. AMR100489-23-F2 was processed using the primary preparative HPLC method described in Table 3. Fractions of Peak ID: AMR100489-23-F2-P2 (Lot No. IN-SDV-D-168-2) (time range of collected fractions: retention time 13.0-13.8 min) were pooled and lyophilized. The final yield of Peak ID: AMR100489-23-F2-P2 (Lot No. IN-SDV-D-168-2) was 1.4 g.

[0334] [Table 15]

[0335] Approximately 109 g of Lot No. LHGE-180125 was processed using the primary preparative HPLC method described in Table 4. Fractions of Peak ID: LHGE-180125-P6 (Lot No. IN-RAS-A-24-6) (time range of collected fractions, retention time 25.5-29.0 min) were pooled and lyophilized. The final yield of Peak ID: LHGE-180125-P6 (Lot No. IN-RAS-A-24-6) was 5.5 g.

[0336] [Table 16]

[0337] Secondary Processing: Approximately 1.4 g of Peak ID: AMR100489-23-F2-P2 (Lot No. IN-SDV-D-168-2) was processed using the preparative HPLC method conditions described in Table 5. The fractions of Peak ID: AMR100489-23-F2-P2-G collected from the secondary processing of # AMR100489-23-F2-P2 (time range of collected fractions, retention time 26.0-27.0 min) were pooled and lyophilized for isolation as described in section 3.8. The final yield of Peak ID: AMR100489-23-F2-P2-G (Lot No. IN-VVP-K-138-7) was 34 mg with a purity of 89.0% (area %).

[0338] [Table 17]

[0339] Peak ID: LHGE-180125-P6 (Lot No. IN-RAS-A-24-6) at approximately 5.5 min was processed using the preparative HPLC method conditions described in Table 6. Fractions of Peak ID: LHGE-180125-P6-B (time range of collected fractions, retention time 5.0 - 5.50 min) were pooled and lyophilized. The final yield of Peak ID: LHGE-180125-P6-B (Lot No. IN-RAS-A-87-2) was 800 mg.

[0340] [Table 18]

[0341] Tertiary Processing: Approximately 34 mg of Peak ID: AMR100489-23-F2-P2-G (Lot No. IN-VVP-K-138-7) was further processed using the preparative HPLC method conditions described in Table 7. The target fractions had a retention time of 6.3-6.9 minutes. The target fractions were pooled and lyophilized. The final yield of highly pure Peak ID: AMR100489-23-F2-P2-G (Lot No. IN-VVP-K-172) was 13 mg with a purity of 95.5% (area %).

[0342] [Table 19]

[0343] Approximately 800 peak ID: LHGE-180125-P6-B (Lot No. IN-RAS-A-87-2) was processed using the preparative HPLC method conditions described in Table 8. The target fractions had a retention time of 23.0-25.0 min. The target fractions were pooled and lyophilized. The final yield of Peak ID: LHGE-180125-P6-B7 (Lot No. IN-RAS-A-106-7) was 15 mg.

[0344] [Table 20]

[0345] Quaternary Processing: Approximately 15 mg of Peak ID: LHGE-180125-P6-B-7 (Lot No. IN-RAS-A-106-7) was processed using the preparative HPLC method conditions described in Table 9. The target fraction (LHGE-180125-P6-B7-C) had a retention time of 22.0-22.8 minutes. The target fractions were pooled and lyophilized. The final yield of Peak ID: LHGE-180125-P6-B7-C (Lot No. IN-RAS-A-119-3) was 0.9 mg with a purity of 81.3% (area %).

[0346] The collected fractions from the preparative run were pooled and lyophilized using a Labconco Lyopholizer (collector temperature maintained at −44° C. under vacuum).

[0347] [Table 21]

[0348] MS and MS / MS. MS and MS / MS data were generated on a Waters QTof Micro mass spectrometer equipped with an electrospray ionization source. Samples were analyzed by negative ESI. Samples (approximately 0.2 mg) were diluted with 50:50 ACN:HO to a concentration of approximately 0.2 mg / mL and introduced by direct infusion.

[0349] The ESI-TOF mass spectrum obtained by injecting a sample of CC-00520 showed [M-H] at m / z 1417.6843. - The ion [MH] was shown. - The mass of the ion is calculated based on the predicted molecular formula C 65 H 110 O 33 was in good agreement with (C 65 H 109 O 33 Calculated value: 1417.6851, error: -0.6 ppm).

[0350] NMR spectroscopy. 1 H NMR, 13 C NMR, 1 H- 1 A series of NMR experiments, including H COSY, HSQC-DEPT, HMBC, ROESY, and 1D TOCSY, were obtained, allowing the assignment of CC-00520.

[0351] [Table 22]

[0352] [Table 23]

[0353] CC-00520 was identified as mogrol-3-O-{[β-D-xylopyranosyl-(1→4)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside}. CC-00520 is related to mogroside V but differs from it by an additional xylose unit attached to GlcIV by forming a 1→4 sugar bond.

[0354] Example 9: Purification and characterization of CC-00539 CC-00539 (mogrol-3-O-{[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[α-L-rhamnopyranosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}) was isolated from a monk fruit (Luo Han Guo) extract containing 90% mogroside V. Isolation of this compound involved multiple workups with preparative HPLC methods, yielding 12 mg (70% purity) of CC-00539. NMR data indicated that the sample contained approximately 70% CC-00539 and approximately 30% mogroside V.

[0355] 1 H, 13 C. 1 H- 1 H COSY, 1 H- 13 C HSQC-DEPT, and 1 H- 13 C HMBC data indicated the presence of a central triterpene core and five sugar units in the structure. The complete structure is 1 H, 13 C. 1 H- 1 H COSY, 1D-TOCSY, 1 H- 13 C HSQC-DEPT and 1 H- 13 C Determined based on HMBC data. 1In addition to the eight methyl protons observed in the central triterpene core, δ H An additional methyl group observed at 1.21 (d, J = 6.2 Hz) indicated the presence of a rhamnose unit in the structure. 1D and 2D NMR data indicated that the remaining four sugar units were glucose. δ using different mixing times (20-140 msec) H 1D-TOCSY data for the anomeric proton of 5.31 reveals that δ H We confirm that the methyl protons at 1.21 belong to the same spin system, and therefore δ H The anomeric proton at 5.31 was assigned to rhamnose H-1. H The anomeric proton of rhamnose observed at 5.31 is Gl cI C-2(δ C 77.4) showed HMBC correlation to Gl cI H-2 (δH 3.49) to the anomeric carbon of rhamnose (δ C The reverse HMBC correlation to 102.0 was also observed, indicating that rhamnose and Gl cI The results were obtained using a 500 MHz NMR instrument equipped with a 2.5 mm inverse probe. 1 In the 1 H NMR spectrum, the rhamnose anomeric protons were observed as a broad singlet, indicating the α-configuration of rhamnose. 13 Before obtaining the C NMR spectrum, use a 500 MHz NMR instrument equipped with a 5 mm broadband probe. 1 When the H NMR spectrum was obtained, the spectrum was better resolved and the rhamnose anomeric protons were observed as a broad doublet with a coupling value of 1.1 Hz, confirming the presence of the α-configuration of rhamnose. 1 H- 1 H COSY and 1 H- 13 C HMBC correlations were used to confirm the sugar linkages in the structure.

[0356] Mass spectrum analysis of CC-00539 by electrospray ionization time-of-flight (ESI-TOF) mass spectrometry in negative polarity mode revealed a [M-H] at m / z 1269.6537. - The ion [MH] was shown. - The mass of the ion is calculated based on the predicted molecular formula C 60 H 102 O 28 was in good agreement with (C 60 H 101 O 28 Calculated value: 1269.6479, error: 4.6 ppm).

[0357] Example 10: Preparation, purification, and characterization of CC-00540 CC-00540 was isolated from the reaction product of the bioconversion of mogroside IIE with α-D-xylosyl fluoride and α-xylosidase. CC-00540 was purified using a preparative HPLC method in multiple replicates to give approximately 3.7 mg with a purity of 95.4% (HPLC area %). A series of 1D and 2D experiments: 1 H, 13 C. 1 H- 1 H COSY, 1 H- 13 C HSQC-DEPT, 1 H- 13 C HSQC-TOCSY, and 1 H- 13 C HMBC was obtained for this sample and the structure was elucidated as mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]. Spectroscopic data indicated the presence of a central triterpene core and four sugar units in the structure: two glucose units and two xylose units. The linkages between the sugar units and their attachment to the central triterpene core were determined based on COSY, TOCSY, and HSQC-DEPT data. Two glucose units exist as β anomers, while two xylose units ( 3 J = 3.1 Hz) exists in the α configuration.

[0358] [Table 24]

[0359] [Table 25]

[0360] Example 11: Preparation, purification, and characterization of CC-00541 CC-00541 was isolated from the reaction product of the bioconversion of mogroside IIE with α-D-xylosyl fluoride and α-xylosidase. CC-00541 was purified using preparative HPLC in multiple replicates to give approximately 11.1 mg with 83.8% purity (HPLC area %), with CC-00540 as the lesser compound. A series of 1D and 2D experiments: 1 H, 13 C. 1 H- 1 H COSY, 1 H- 13 C HSQC-DEPT, 1 H- 13 C HSQC-TOCSY, and 1 H- 13 C HMBC was obtained for this sample and the structure was elucidated as mogrol-3-O-[β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]. Spectroscopic data indicated the presence of a central triterpene core and three sugar units in the structure: two glucose units and one xylose unit. The linkages between the sugar units and their attachment to the central triterpene core were determined based on COSY, TOCSY, and HSQC-DEPT data. Two glucose units exist as β anomers, while the remaining xylose unit ( 3 J = 3.6 Hz) exists in the α configuration.

[0361] [Table 26]

[0362] [Table 27]

[0363] Example 12: Preparation, purification, and characterization of CC-00542 CC-0542 was isolated from the reaction product of the bioconversion of mogroside IIE with α-D-xylosyl fluoride and α-xylosidase. CC-00542 was purified using preparative HPLC in multiple replicates to give approximately 6.8 mg with 84% purity (HPLC area %). A series of 1D and 2D experiments: 1 H, 13 C. 1 H- 1 H COSY, 1 H- 13 C HSQC-DEPT, 1 H- 13 C HSQC-TOCSY, and 1 H- 13 C HMBC was obtained for this sample and the structure was elucidated as mogrol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-β-D-glucopyranoside. Spectroscopic data indicated the presence of a central triterpene core and three sugar units: two glucose units and one xylose unit. The linkages between the sugar units and their attachment to the central triterpene core were determined based on COSY, TOCSY, and HSQC-DEPT data. Two glucose units exist as the β anomer, while the remaining xylose unit ( 3 J = 3.6 Hz) exists in the α configuration.

[0364] [Table 28]

[0365] [Table 29]

[0366] Example 13: Preparation, purification, and characterization of CC-00550 [ka] 3-O-β-D-Glucopyranosylmogrol 24-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (4). To a solution of 3 (613.8 mg, 0.2625 mmol) in dry THF (11 mL) and dry MeOH (11 mL) was added dropwise 0.5 M NaOMe in MeOH (0.52 mL, 0.2625 mmol) at room temperature under a N atmosphere. The mixture was stirred at the same temperature for 18 h, neutralized with 10% aqueous AcOH, and concentrated under reduced pressure. The remaining white solid was purified by preparative HPLC (Phenomenex Luna C18(2) column, 21.2 × 250 mm, 5 μm particles, 25% CH3CN in H2O, 20 mL / min, t R =10.2 min). The combined fractions were concentrated under reduced pressure and the residue was dried under vacuum to give the title compound 4 (225.5 mg, 77%) as a white powder. f = 0.68 (silica gel, CH2Cl2 / MeOH / H2O = 5:4:1). 1H NMR(500MHz,ピリジン-d5)δ 7.68(br s,1H),7.50(br s,1H),7.21-6.87(m,4H),6.63(br s,2H),6.55(br s,1H),6.42(s,1H),6.36(br s,1H),6.20(br s,1H),5.73(s,1H),5.67(br s,1H),5.52(d,J=5.7Hz,1H),5.02(d,J=7.8Hz,1H),4.92(d,J=7.8Hz,2H),4.87-4.82(m,1H),4.82-4.74(m,2H),4.62(dd,J=9.2,3.3Hz,1H),4.58-4.47(m,2H),4.45-4.34(m,2H),4.34-4.27(m,2H),4.27-4.19(m,5H),4.19-4.14(m,1H),4.14-4.08(m,1H),4.08-4.01(m,2H),4.01-3.87(m,5H),3.70(s,1H),3.01-2.89(m,1H),2.84-2.72(m,1H),2.53-2.41(m,1H),2.39-2.26(m,1H),2.21-1.92(m,7H),1.72(d,J=6.2Hz,3H),1.73-1.65(m,3H),1.63(d,J=7.5Hz,1H),1.59(s,3H),1.60-1.53(m,2H),1.53-1.43(m,1H),1.49(s,3H),1.38(s,3H),1.33(s,3H),1.15(s,3H),1.13-1.05(m,1H),1.09(d,J=4.3Hz,3H),1.05-0.96(m,1H),0.89(s,3H),0.81(s,3H). 13C NMR (125 MHz, pyridine-d5) δ 144.7, 118.9, 107.9, 105.3, 103.9, 102.7, 90.0, 88.4, 79.9, 79.2, 79.0, 78.7, 78.6, 78.2, 77.1, 77.0, 76.0, 75.8, 74.5, 73.1, 73.0, 72.8 (2C), 72.2, 71.9, 70.4, 69. 9, 63.5, 63.0, 52.0, 50.1, 47.9, 43.9, 42.9, 41.6, 40.6, 37.3, 36.5, 34.9, 33.4, 30.1, 30.0, 29.2, 28.2, 27.5, 27.3, 26.9, 26.7, 25.12, 25.06, 19.7, 19.3, 19.1, 17.6.

[0367] Example 14: Preparation, purification, and characterization of CC-00551 CC-00551 was isolated from a monk fruit (Luo Han Guo) extract containing 90% mogroside V. Analysis of CC-00551's 1D and 2D NMR and MS spectra allowed the complete assignment of its structure to mogrol-3-O-[{β-D-glucopyranosyl-(1→6)}-β-D-glucopyranoside]-24-O-[{β-D-glucopyranosyl-(1→2)}-{β-D-xylopyranosyl-(1→6)}-β-D-glucopyranoside]. The central triterpene core was assigned as follows: 1 H, 13 C. 1 H- 1 H COSY, 1 H- 13 C HSQC-DEPT and 1 H- 13 A combination of C HMBC data and a series of 1D-TOCSY experiments, combined with 1D and 2D NMR data, were used to assign and link the sugar units. Evaluation of the NMR data led to the conclusion that this compound has four glucose units and one xylose unit. The xylose unit is linked to GlcI by a 1→6 sugar bond.

[0368] The ESI-TOF mass spectrum obtained by injection of a sample of CC-00551 showed [M-H] at m / z 1255.6377.- The ion [MH] was shown. - The mass of the ion is calculated based on the predicted molecular formula C 59 H 100 O 28 was in good agreement with (C 59 H 99 O 28 Calculated value: 1255.6323, error: 4.3 ppm). MS data indicates the molecular formula C 59 H 100 O 28 The ions observed at m / z 1291.5961 and 1353.5873 were likely [M-H+HCl] ions, respectively. - and [M-H+H3PO4] - The MS / MS spectrum shows [M−H] at m / z 1255.6. - The ion was selected for fragmentation and showed the loss of a xylose unit at m / z 1123.5693, followed by the sequential loss of four glucose units at m / z 961.5647, 799.5051, 637.4304, and 475.3798, indicating the presence of one xylose and four glucose units in the structure.

[0369] [Table 30]

[0370] [Table 31]

[0371] [Table 32]

[0372] Example 15: Sensory analysis The following samples were tested at 400 ppm in water at 4°C. Sample size was limited; therefore, the number of panelists per sample ranged from 1 to 10, and the volume tested ranged from 2 to 10 mL. The sweetness of each sample was compared to a sucrose standard.

[0373] [Table 33]

[0374] Example 16: Preparation of CC-00497 CC-00497 was prepared by bioconversion of isomogroside V. 250 mg of isomogroside V and 10 mg of β-galactosidase G5160 in 3.1 mL of pH 5 sodium acetate buffer were stirred at 37°C for 3 days and heated for 30 minutes. The crude mixture was directly purified using preparative HPLC to give 2.8 mg of CC-00497. The structure was confirmed by 1D and 2D NMR analysis.

[0375] [Table 34]

[0376] [Table 35]

[0377] [Table 36]

Claims

1. The following structure: 【Chemical 1】 【Chemistry 2】 【Chemistry 3】 A mogroside selected from

2. 10. The mogroside of claim 1, which is isolated and purified.

3. 10. A composition comprising the mogroside of claim 1 and at least one other substance, the composition comprising at least 5% by weight of the mogroside of claim 1.

4. 10. A consumable product comprising the mogroside of claim 1 or the composition of claim 3.

5. 5. The consumable product of claim 4, which is a beverage or beverage product.

6. 6. The consumable of claim 5, wherein the consumable is a beverage, and the beverage comprises the mogrosides at a concentration of 1 ppm to 100 ppm.

7. 6. The consumable product of claim 5, wherein the beverage further comprises at least one additional sweetener selected from the group consisting of carbohydrate sweeteners, rare sugar sweeteners, high-potency sweeteners, synthetic sweeteners, and combinations thereof.

8. 6. The consumable product of claim 5, wherein the beverage further comprises at least one additional sweetener selected from the group consisting of rebaudioside A, rebaudioside M, rebaudioside D, mogroside V, siamenoside I, and mogrol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}.

9. 6. The consumable product of claim 5, wherein the beverage further comprises at least one functional ingredient selected from the group consisting of saponins, antioxidants, dietary fiber sources, fatty acids, vitamins, glucosamine, minerals, preservatives, hydration agents, probiotics, prebiotics, weight management agents, osteoporosis management agents, phytoestrogens, long chain primary aliphatic saturated alcohols, phytosterols, and combinations thereof.

10. 6. The consumable product of claim 5, wherein the beverage further comprises at least one additive selected from the group consisting of carbohydrates, polyols, amino acids and their corresponding salts, polyamino acids and their corresponding salts, sugar acids and their corresponding salts, nucleotides, organic acids, inorganic acids, organic salts including organic acid salts and organic base salts, inorganic salts, bitter compounds, caffeine, flavoring agents and flavor components, astringent compounds, proteins or protein hydrolysates, surfactants, emulsifiers, plant extracts, flavonoids, alcohols, polymers, and combinations thereof.

11. 6. The consumable product of claim 5, wherein the beverage is selected from the group consisting of frozen carbonated beverages, enhanced sparkling beverages, cola, fruit-flavored sparkling beverages, ginger ale, soft drinks and root beer, fruit juice, fruit-flavored juice, juice drink, nectar, vegetable juice, vegetable-flavored juice, sports drink, energy drink, enhanced water drink, vitamin-enhanced water, near-water drink, coconut water, tea-based beverage, coffee, cocoa drink, dairy beverage, cereal extract beverage, protein drink, yogurt drink, and smoothie.

12. 6. The consumable product of claim 5, wherein the beverage is a low-calorie or mid-calorie beverage.

13. 6. The consumable product of claim 5, wherein the consumable product is a beverage product, and the beverage product comprises the mogrosides at a concentration of 10 ppm to 1,000 ppm.

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