Mogroside and uses thereof

Abstract

Provided herein are mogrosides containing non-glucose glycosides. Also provided are compositions, including consumables, comprising the mogrosides described herein.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Application No. 62 / 810,556, filed February 26, 2019, the entire contents of which are incorporated herein by reference.

[0003] field

[0004] This disclosure relates to mogroside compounds containing one or more non-glucose carbohydrates and methods for their production. More specifically, this disclosure relates to the use of mogroside compounds containing one or more non-glucose carbohydrates as sweeteners in consumer products.

[0005] background

[0006] Monk fruit extract, obtained from *Siraitia grosvenori* (a plant in the Cucurbitaceae family), is commercially used as a natural sweetener. However, monk fruit extracts may have taste characteristics that hinder their use as alternatives to caloric sweeteners (e.g., sugar) in food and beverage compositions. For example, the extract may have certain off-flavors or lingering aftertastes, or it may take longer than desired for sweetness to develop after consumption (i.e., delayed sweetness onset).

[0007] There is still a need for sweeteners with reduced calorie content, which are low-calorie or calorie-free, have improved taste characteristics, and for foods and beverages containing said sweeteners.

[0008] Overview

[0009] Typical mogrosides are characterized by glucose-containing glycosides at the C-3 and C24 positions of the mogroside core. This disclosure relates to novel mogroside compounds that contain at least one carbohydrate other than glucose at the C3 and C24 positions of the glycoside.

[0010] In one aspect, the present invention provides a mogroside of formula I:

[0011]

[0012] in:

[0013] when When it is a double bond, X is O;

[0014] when When it is a single bond, X is selected from OH and H, and the carbon attached to X is replaced by H to provide the appropriate valence (e.g., HC[(CH2)(C)]-X);

[0015] R 1R 2 R 3 R 4 R 5 and R 6 Each is independently selected from hydrogen-rich, pentose, or hexose carbohydrates, where R 1 -R 6 At least one of them is a non-glucose pentose or hexose carbohydrate; and

[0016] The total number of carbohydrates is four, five, or six.

[0017] 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, lyseose, fucose, allose, allusose, azoose, mannose, gulose, iodose, galactose, tarose, and rhamnose. The carbohydrate bond can be α-configured or β-configured. The carbohydrate can be D-configured or L-configured.

[0018] In one embodiment, R 1 -R 4 One is a non-glucose pentose or hexose carbohydrate, and R 5 and R 6 It is hydrogen.

[0019] In another embodiment, R 1 -R 4 One of them is xylose, and R 5 and R 6 It is hydrogen.

[0020] In some embodiments, the total number of carbohydrates in the compound of Formula I is four, five, or six, and the non-glucose pentose or hexose carbohydrate is xylose.

[0021] In some other embodiments, the total number of carbohydrates in the compound of Formula I is four, five, or six, and the non-glucose pentose or hexose carbohydrates are not xylose.

[0022] In a second aspect, the present invention provides a mogroside of formula II:

[0023]

[0024] in:

[0025] when When it is a double bond, X is O;

[0026] when When it is a single bond, X is selected from OH and H, and the carbon attached to X is replaced by H to provide the appropriate valence (e.g., HC[(CH2)(C)]-X);

[0027] R 1 R 2 R 3 R 4 and R 7 Each is independently selected from hydrogen-rich, pentose, or hexose carbohydrates, where R 1 R 2 R 3 R 4 and R 7 At least one of them is a non-glucose pentose or hexose carbohydrate.

[0028] 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, lyseose, fucose, allose, allusose, azoose, mannose, gulose, idose, galactose, tarose, and rhamnose. The carbohydrate bond can be α-configured or β-configured.

[0029] In some embodiments, when the total number of carbohydrates in the compound of Formula II is three, four, five, or six, the non-glucose pentose or hexose carbohydrate is xylose.

[0030] The specific mogrosides disclosed herein include, but are not limited to, mogroside-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-xylanosyl-(1→6)]-β-D-glucopyranoside}, mogroside-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-L-rhamnosyl-(1→6)]-β-D-glucopyranoside}, mogroside-3-O-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[β-D-xylanosyl-(1→6)-β-D-glucopyranoside]-24-O-[β-D-xylanosyl-(1→6)-β-D-glucopyranoside]. [Glucoside], mogroside-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-D-galactopyranosyl-(1→6)]-β-D-glucopyranoside}, mogroside-3-O-[α-L-rhamnosyl-(1→6)-β-D-glucopyranoside]-24-O-[β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside], mogroside-3-O-[{β-D-glucopyranosyl-(1→6)}-β-D-glucopyranoside]-24-O-[{β-D-xylosyl-(1→2)}-{β-D-glucopyranosyl-(1→6)] [α-D-xylopyranoside], mongholic acid-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside, mongholic acid-3-O-{[β-D-xylopyranosyl-(1→4)]-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}, mongholic acid-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}, mogroside-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside], mogroside-3-O-[β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside], mogroside-3-O-[α-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside], mogroside-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-β-D-glucopyranoside,3-O-β-D-glucopyranosylmogroside, 24-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside, and mogroside-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}.

[0031] In one specific embodiment, mogrosides are isolated and purified.

[0032] In another aspect, the present invention is a composition comprising at least one mogroside described herein. In one specific embodiment, the present invention is a composition comprising at least one isolated and purified mogroside described herein.

[0033] In one embodiment, the present invention is a sweetener composition comprising at least one mogroside described herein.

[0034] In another embodiment, the present invention is a flavor-enhancing composition comprising at least one mogroside described herein, wherein when added to a consumer product, the mogroside is present in the composition in an amount that effectively provides a concentration equal to or below the flavor recognition threshold of the mogroside.

[0035] In yet another embodiment, the present invention is a sweetness-enhancing composition comprising at least one mogroside described herein, wherein when the sweetness-enhancing composition is added to a consumer product, the mogroside is present in the composition in an amount that effectively provides a concentration equal to or below the sweetness recognition threshold of the mogroside.

[0036] In yet another embodiment, the present invention is a consumer product comprising at least one mogroside described herein. Suitable consumer products include, but are not limited to, liquid-based or dry-based consumer products, such as pharmaceutical compositions, edible gel mixtures and compositions, dental compositions, food, beverages, and beverage products.

[0037] In one specific embodiment, the present invention is a beverage comprising at least one mogroside described herein. In one specific embodiment, the mogroside is present in the beverage at a concentration higher than, equal to, or lower than the threshold sweetness identification concentration of the mogroside.

[0038] In another aspect, the present invention is a method for preparing a consumer product, the method comprising (i) providing a consumer product matrix and (ii) adding at least one mogroside described herein to the consumer product matrix to provide a consumer product.

[0039] In one specific embodiment, the present invention is a method for preparing a beverage, the method comprising (i) providing a beverage matrix and (ii) adding at least one mogroside described herein to the beverage matrix to provide a beverage.

[0040] In another aspect, the present invention is a method for enhancing the sweetness of a consumer product, the method comprising (i) providing a consumer product containing at least one sweetening ingredient and (ii) adding at least one isolated and purified mogroside described herein to the consumer product to provide a consumer product with enhanced sweetness, wherein the mogroside is present in the consumer product with enhanced sweetness at a concentration equal to or below a sweetness recognition threshold of the mogroside. In one specific embodiment, the consumer product is a beverage.

[0041] In some embodiments, the compositions of the present invention comprise one or more sweeteners, additives, and / or functional ingredients.

[0042] In one embodiment, the present invention is a consumer product comprising at least one mogroside of the present invention and one or more sweeteners, additives, and / or functional ingredients. In another embodiment, the present invention is a beverage comprising at least one mogroside of the present invention and one or more sweeteners, additives, and / or functional ingredients.

[0043] In another aspect, the present invention is a method for purifying mogrosides of the present invention, the method comprising (i) passing a solution containing a source material comprising mogrosides of the present invention through an HPLC column, and (ii) eluting a fraction containing mogrosides of the present invention to provide a purified mogroside composition comprising at least about 80% by weight of mogrosides of the present invention. Exemplary source materials include, but are not limited to, mixtures of mogrosides, Luo Han Guo extracts (commercial or prepared), and compositions produced by the biotransformation processes described herein. Attached Figure Description

[0044] Figure 1 shows the LC-MS of 90% mogroside V starting material (MV 90) (Example 1).

[0045] Figure 2 The HPLC trace of the reaction between MogIIIE and rhamnose (Example 2) is shown.

[0046] Figure 3 shows the peak 45 (separated by the reaction of MogIIIE and rhamnose (Example 2)). Figure 3A ) and 46 ( Figure 3B Maldi mass spectrometry.

[0047] Figure 4 The HPLC trace of the reaction between MogIIIE and galactose (Example 2) is shown.

[0048] Figure 5 shows the peak 108 separated by the reaction of MogIIIE and galactose (Example 2). Figure 5A ) and 109 ( Figure 5B Maldi mass spectrometry.

[0049] Figure 6 MS traces of the reaction between MogIIIE and xylose (Example 2) are shown.

[0050] Figure 7 The Maldi mass spectrum of peak 39, separated by the reaction of MogIIIE and xylose (Example 2), is shown. Detailed Implementation

[0051] This document discloses compounds, compositions, and methods for use as sweeteners in beverages and foods. The disclosed compounds include mogroside compounds containing non-glucose carbohydrate units. Surprisingly, the mogrosides of the present invention have been found to be usable in a variety of applications, such as as sweeteners or food additives in consumer products like beverages and foods. Such compositions include the mogrosides of the present invention, alone or in blends with other sweeteners, additives, or flavor enhancers.

[0052] I. Definition

[0053] As used herein, the term "consumable" means a substance suitable for individual ingestion. Exemplary consumer products include, but are not limited to, edible gel mixtures and compositions, dental compositions, food (sweeteners, condiments, chewing gum, cereal compositions, baked goods, dairy products, and tabletop sweetener compositions), beverages, and beverage products.

[0054] As used in this article, the term "monk fruit" or "Luo Han Guo" (luo hanguo) refers to the fruit of *Siraitia grosvenori*, a member of the Cucurbitaceae family.

[0055] As used herein, the term "pH" refers to a number on a logarithmic scale representing the acidity or alkalinity of a solution, where 7 is neutral, lower values ​​are more acidic, and higher values ​​are more alkaline. pH equals -log10 c, where c is the concentration of hydrogen ions in moles per liter.

[0056] As used herein, the term "purified" means that the purity of a compound has been increased, so that it exists in a purer form than it does in its natural environment and / or extracts. Purity is a relative term and does not necessarily refer to absolute purity.

[0057] II.Compounds

[0058] This disclosure provides a mogroside of formula I:

[0059]

[0060] in:

[0061] when When it is a double bond, X is O;

[0062] when When it is a single bond, X is selected from OH and H, and the carbon attached to X is replaced by H to provide the appropriate valence (e.g., HC[(CH2)(C)]-X);

[0063] R 1 R 2 R 3 R 4 R 5 and R 6 Each is independently selected from hydrogen-rich, pentose, or hexose carbohydrates, where R 1 -R 6 At least one of them is a non-glucose pentose or hexose carbohydrate; and

[0064] The total number of carbohydrates is four, five, or six.

[0065] 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, lyseose, fucose, allose, allusose, azoose, mannose, gulose, idose, galactose, tarose, and rhamnose. The carbohydrate bond can be α-configured or β-configured. The carbohydrate can be D-configured or L-configured.

[0066] The compound of Formula I has at least four total carbohydrates (i.e., the sum of carbohydrate units at positions C-3 and C-24). 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 still another embodiment, the compound of Formula I has six total carbohydrates.

[0067] In one embodiment, R 1 -R 4 One is a non-glucose pentose or hexose carbohydrate, and R 5 and R 6 It is hydrogen.

[0068] In one embodiment, the total number of carbohydrates in the compound of Formula I is four, five, or six, and the non-glucose pentose or hexose carbohydrate is xylose.

[0069] In another embodiment, the total number of carbohydrates in the compound of Formula I is four, five, or six, and the non-glucose pentose or hexose carbohydrate is rhamnose.

[0070] In yet another embodiment, the total number of carbohydrates in the compound of Formula I is four, five, or six, and the non-glucose pentose or hexose carbohydrate is galactose.

[0071] In some embodiments, the mogroside of formula I is 11-oxomogroside of formula Ia:

[0072]

[0073] Where R 1 R 2 R 3 R 4 R 5 and R 6 As defined above for Equation I.

[0074] In some other embodiments, the mogroside of formula I is 11-deoxymogroside of formula Ib:

[0075]

[0076] Where R 1 R 2 R 3 R 4 R 5 and R 6 As defined above for Equation I.

[0077] In some other embodiments, mogroside of formula I is a compound of formula Ic:

[0078]

[0079] Where R 1 R 2 R 3 R 4 R 5 and R 6 As defined above for Equation I.

[0080] 11-OH can be in the R-configuration or the S-configuration.

[0081] This disclosure also provides a mogroside of formula II:

[0082]

[0083] in:

[0084] when When it is a double bond, X is O;

[0085] when When it is a single bond, X is selected from OH and H, and the carbon attached to X is replaced by H to provide the appropriate valence (e.g., HC[(CH2)(C)]-X);

[0086] R 1 R 2 R 3 R 4 and R 7 Each is independently selected from hydrogen-rich, pentose, or hexose carbohydrates, where R 1 R 2 R 3 R 4 and R 7 At least one of them is a non-glucose pentose or hexose carbohydrate.

[0087] 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, lyseose, fucose, allose, allusose, azoose, mannose, gulose, idose, galactose, tarose, and rhamnose. The carbohydrate bond can be α-configured or β-configured.

[0088] The compound of Formula II has at least three total carbohydrates (i.e., the sum of carbohydrate units at positions C-3 and C-24). 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.

[0089] In one embodiment, the total number of carbohydrates in the compound of Formula II is three, four, five, or six, and the non-glucose pentose or hexose carbohydrate is xylose.

[0090] In another embodiment, the total number of carbohydrates in the compound of formula II is three, four, five, or six, and the non-glucose pentose or hexose carbohydrate is rhamnose.

[0091] In some embodiments, the mogroside of formula II is 11-oxomogroside of formula IIa:

[0092]

[0093] Where R 1 R 2 R 3 R 4 and R 7 As defined above for Equation II.

[0094] In some other embodiments, the mogroside of formula II is 11-deoxymogroside of formula IIb:

[0095]

[0096]

[0097] Where R 1 R 2 R 3 R 4 and R 7 As defined above for Equation II.

[0098] In some other embodiments, the mogroside of formula II is a compound of formula IIc:

[0099]

[0100] Where R 1 R 2 R 3 R 4 and R 7 As defined above for Equation II.

[0101] 11-OH can be in the R-configuration or the S-configuration.

[0102] In one embodiment, the mogroside of the present invention is selected from the following:

[0103] (i) Monk fruit alcohol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[β-D-xylopyranosyl-(1→6)]-β-D-glucopyranoside}(CC-00489):

[0104]

[0105] (ii) Monk fruit alcohol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-L-rhamnosyl-(1→6)]-β-D-glucopyranoside} (CC-00491):

[0106]

[0107] (iii) Monk fruit alcohol-3-O-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside](CC-00497):

[0108]

[0109] (iv) Monk fruit alcohol-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranosyl-(1→2)]-[α-D-galactopyranosyl-(1→6)]-β-D-glucopyranoside} (CC-00498):

[0110] as well as

[0111] (v) Monk fruit alcohol-3-O-[α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside](CC-00500):

[0112]

[0113] (vi) Monk fruit alcohol-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):

[0114]

[0115] (vii) Monk fruit alcohol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside](CC-00518)

[0116]

[0117] (viii) Monk fruit alcohol-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)

[0118]

[0119] (ix) Monk fruit alcohol-3-O-{[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[α-L-rhamnosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}(CC-00539)

[0120]

[0121] (x) Monk fruit alcohol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside](CC-00540)

[0122]

[0123] (xi) Monk fruit alcohol-3-O-[β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside](CC-00541)

[0124]

[0125] (xii) Monk fruit alcohol-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-β-D-glucopyranoside (CC-00542)

[0126]

[0127] (xiii) 3-O-β-D-glucopyranosylmogrostrol 24-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (CC-00550)

[0128] as well as

[0129] (xiv) Monk fruit alcohol-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)

[0130]

[0131] In an exemplary embodiment, the mogroside of the present invention is isolated and purified.

[0132] As used herein, the term "isolated and purified" means that the compound is about 95% or more by weight based on dry weight, i.e., a purity greater than 95%. The remainder of the mixture is typically other mogrosides and / or mogroside extracts. In more specific embodiments, the mogrosides of the formula described herein have a purity of about 96% or more, about 97% or more, about 98% or more, or about 99% or more. In some embodiments, the mogrosides are enzymatically produced.

[0133] In some embodiments, the mogrosides of the present invention are sweet. The sweetness of a given composition is typically measured with reference to a sucrose solution. See also "A Systematic Study of Concentration-Response Relationships of Sweeteners", GE DuBois, DE Walters, S.S. Schiffman, Z.S. Warwick, B.J. Booth, S.D. Pecore, K. Gibes, B.C. Arr, and L.M. Brands, in Sweeteners: Discovery, Molecular Design and Chemoreception, edited by DE Walters, F.T. Orthoefer, and GE DuBois, American Chemical Society, Washington, DC (1991), pp. 261-276.

[0134] The sweetness of non-sucrose sweeteners can be measured relative to a sucrose reference by determining the sucrose equivalent (SE) of the non-sucrose sweetener. Typically, members of a taste expert panel are trained to detect the sweetness of a reference sucrose solution containing between 1% and 15% sucrose (w / v). Other non-sucrose sweeteners are then tasted at a series of dilutions to determine the concentration of a non-sucrose sweetener that is as sweet as a given percentage of the sucrose reference. 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.

[0135] In one embodiment, mogroside is present in an amount that, when added to a consumer product, provides a sucrose equivalent of more than about 2% (w / v), such as 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% SE.

[0136] The amount of sucrose in a reference solution can be described using Brix (°Bx), and thus another measure of sweetness. One Brix is ​​1 gram of sucrose in 100 grams of solution, and represents the strength of said solution as a weight percentage (%w / w) (strictly speaking, by mass). In one embodiment, the mogroside of the present invention, when added to a consumer product, is present in an amount providing a sweetness equivalent of from about 0.50 to 14 Brix, such as from about 5 to about 12 Brix, about 7 to 10 Brix, or above 10 Brix.

[0137] In an exemplary embodiment, the isolated and purified mogrosides of the present invention have a sweetness that is about 30% or more higher than that of partially purified mogrosides or mogrosides, for example, such as about 40% or more, about 50% or more, about 60% or more, about 70% or more, about 80% or more, or about 90% or more.

[0138] In other exemplary embodiments, the isolated and purified mogrosides of the present invention have a bitterness (a taste stimulated by certain substances such as quinine, caffeine, and sucrose octaacetate) that is at least about 30%, for example, 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%, lower than that of partially purified mogrosides or mogrosides. In one specific embodiment, the isolated and purified mogrosides of the present invention are substantially free of bitterness. Methods for measuring the bitterness of compounds are known in the art.

[0139] In other exemplary embodiments, the isolated and purified mogrosides of the present invention have a residual sweet aftertaste (sweetness intensity after regurgitation) that is at least 30% lower than that of partially purified mogrosides or mogrosides, for example, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90%. In one specific embodiment, the isolated and purified mogrosides of the present invention have virtually no residual sweet aftertaste. Methods for measuring residual sweet aftertaste are known in the art.

[0140] In other exemplary embodiments, the isolated and purified mogrosides of the present invention have a metallic taste (a taste associated with metallic tin or iron) that is at least 30% lower, for example, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% lower than that of partially purified mogrosides or mogrosides. In one specific embodiment, the isolated and purified mogrosides of the present invention have essentially no metallic taste.

[0141] In exemplary embodiments, compared to partially purified mogrosides or mogrosides, the isolated and purified mogrosides of the present invention exhibit a maximum response (maximum sweetness (%SE) reached with increasing compound concentration) that is at least about 30% higher, such as 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% higher. Methods for measuring the maximum response of the compound are known in the art. In one embodiment, the method is an in vitro cell assay. In some embodiments, the cells express sweet taste receptors or dimers of sweet taste receptors.

[0142] In other exemplary embodiments, the isolated and purified mogrosides of the present invention exhibit a sweetness onset (the time to experience maximum sweetness) that is at least about 30% shorter than that of partially purified mogrosides or mogrosides, for example, 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% shorter. Methods for measuring the sweetness onset are known in the art. In one embodiment, the method is an in vitro cell assay. In some embodiments, the cells express sweet taste receptors or dimers of sweet taste receptors.

[0143] III. Composition

[0144] This invention includes compositions containing at least one mogroside of the present invention. As used herein, the term "composition" refers to a mixture of at least one mogroside of the present invention and at least one other substance.

[0145] In one specific embodiment, the at least one other substance does not exist in nature with the diterpene glycoside and / or does not mix with the diterpene glycoside. Therefore, these compositions do not exist in nature.

[0146] In one embodiment, the present invention is a composition comprising at least one mogroside of the present invention, said mogroside being provided as part of a mixture. In a specific embodiment, the mixture is selected from the group consisting of mogroside, mogroside, byproducts of other mogroside separation and purification processes, commercially available mogroside extracts, byproducts of biotransformation reactions, or any combination thereof.

[0147] In one embodiment, the mixture contains at least one mogroside of the present invention in an amount ranging from about 1% to about 99% on a dry basis by weight, such as about 5% to about 99%, from about 10% to about 99%, from about 20% to about 99%, from about 30% to about 99%, from about 40% to about 99%, from about 50% to about 99%, from about 60% to about 99%, from about 70% to about 99%, from about 80% to about 99%, and from about 90% to about 99%. In a specific embodiment, the mixture contains at least one mogroside of the present invention in an amount greater than about 90% on a dry basis by weight, such as 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%.

[0148] The composition may contain at least about 5% by weight, such as 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% of the mogrosides of the present invention.

[0149] In other embodiments, the composition has a total mogroside content of about 95% or more on a dry basis by weight. In some embodiments, the composition has a total mogroside content of about 96% or more, about 97% or more, about 98% or more, or about 99% or more. As used herein, “total mogroside content” refers to the sum of the relative weight contributions of each mogroside in the sample, including non-glucose-substituted mogrosides as described herein.

[0150] As used herein, the term "purified mogrosides" refers to mogrosides present in a mixture (e.g., monk fruit) at a weight of at least about 50%, such as 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%.

[0151] In one embodiment, the present invention is a composition comprising at least one mogroside described herein, wherein the mogroside is provided as a pure compound, i.e., >99% by weight on a dry basis.

[0152] The mogrosides of the present invention can be effectively provided in the composition at concentrations ranging from about 1 ppm to about 10,000 ppm, such as from about 1 ppm to about 4,000 ppm, from about 1 ppm to about 3,000 ppm, from about 1 ppm to about 2,000 ppm, from about 1 ppm to about 1,000 ppm, from about 1 ppm to about 600 ppm, from about 1 ppm to about 500 ppm, from about 1 ppm to about 400 ppm, from about 1 ppm to about 300 ppm, from about 1 ppm to about 200 ppm, or from about 1 ppm to about 100 ppm when the composition is added to a consumer product.

[0153] In another embodiment, the mogroside of the present invention is present in the composition in an amount that effectively provides mogroside concentrations 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, and from about 100 ppm to about 1,000 ppm.

[0154] 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 ranges from about 500:1 to about 2:1, 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 2 ... From approximately 1 to approximately 100:1, from approximately 500:1 to approximately 50:1, from approximately 500:1 to approximately 25:1, from approximately 500:1 to approximately 10:1, from approximately 400:1 to approximately 300:1, from approximately 400:1 to approximately 200:1, from approximately 400:1 to approximately 100:1, from approximately 400:1 to approximately 50:1, from approximately 400:1 to approximately 25:1, from approximately 400:1 to approximately 10:1, from approximately 400:1 to approximately 6.67:1. From about 300:1 to about 200:1, from about 300:1 to about 100:1, from about 300:1 to about 50:1, from about 300:1 to about 25:1, from about 300:1 to about 10:1, from about 300:1 to about 6.67:2, from about 200:1 to about 100:1, from about 200:1 to about 50:1, from about 200:1 to about 25:1, from about 200:1 to about 10:1, from about 100:1 to about 50:1, from about 100:1 to about 25:1, from about 100:1 to about 10:1, from about 100:1 to about 6.67:1, from about 50:1 to about 25:1, from about 50:1 to about 25:1, from about 50:1 to about 10:1, from about 50:1 to about 6.65:1, from about 25:1 to about 10:1, from about 25:1 to about 6.67:1, from about 10:1 to about 6.67:1, and any range in between.

[0155] A. Sweetener composition

[0156] As described above, in some embodiments, the mogrosides of the present invention are sweet. Therefore, the present invention also provides sweetener compositions comprising at least one mogroside of the present invention. As used herein, the term "sweetener composition" refers to a mixture of at least one mogroside of the present invention and at least one other substance.

[0157] In one specific embodiment, the at least one other substance does not exist in nature with and / or is not blended with the mogroside. Therefore, these sweetener compositions do not exist in nature. In one embodiment, the at least one other substance modifies the taste characteristics of the at least one mogroside to provide a composition with a taste characteristic more like sucrose compared to the mogroside in nature and (if applicable) the at least one other substance in nature. For example, in some embodiments, the composition exhibits one or more of the following characteristics: improved sweetness potency, improved mouthfeel, shortened sweetness retention, reduced bitterness, and / or reduced metallic taste.

[0158] In one aspect, the sweetener composition comprises a sweetening amount of mogroside of the present invention. As used herein, “sweetening amount” refers to the amount of compound required to provide detectable sweetness when present in a consumer product (e.g., a beverage) and is also referred to as the “sweetness detection threshold.”

[0159] In one embodiment, when added to a sweetening composition or a sweetening consumer product, the sweetener composition provides a sucrose equivalent of more than about 2% (w / v), such as 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%.

[0160] In some embodiments, the sweetener composition comprises at least about 5% by weight, such as 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% of the mogroside of the present invention.

[0161] In some embodiments, the sweetener composition comprises at least 5% by weight of at least one other substance (e.g., a sweetener), such as 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%.

[0162] In some embodiments, the at least one other substance is a sweetener, i.e., the sweetener composition is a sweetener blend or contains a sweetener blend. When added to a sweetener composition or a sweetener consumer product, such a sweetener composition can provide a sucrose equivalent of more than about 2% (w / v), such as 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%.

[0163] In one embodiment, the at least one other substance is a sweetener different from the mogroside of the present invention. The sweetener may be present in a sweetening amount.

[0164] In one embodiment, the at least one other substance is a natural high-efficiency sweetener. As used herein, the phrase "natural high-efficiency sweetener" means any sweetener found in plants or other species that requires no further purification and has a sweetness potency greater than sucrose, fructose, or glucose, while having fewer calories. Natural high-efficiency sweeteners may be provided as pure compounds or alternatively as part of an extract.

[0165] In another embodiment, at least one other substance is a synthetic sweetener. As used herein, the phrase "synthetic sweetener" means any composition that is not naturally found in nature and has a sweetening potency greater than that of sucrose, fructose, or glucose, but has fewer calories.

[0166] In other embodiments, combinations of natural high-efficiency sweeteners and synthetic sweeteners are considered.

[0167] In other embodiments, the at least one other substance is a carbohydrate sweetener. Suitable carbohydrate sweeteners are selected from, but are not limited to, the group consisting of: sucrose, glyceraldehyde, dihydroxyacetone, erythrose, threose, erythulose, arabinose, lysose, ribose, xylose, ribulose, xylulose, allose, azoose, galactose, glucose, gulose, idole, mannose, tarose, fructose, allulose, sorbose, tagatose, mannoheptulose, sedoheptulose, octanose, fucose, rhamnose, arabinose, mesobiose, sialic acid, and combinations thereof.

[0168] Other suitable sweeteners include: rebaudin A, rebaudin B, rebaudin C, rebaudin D, rebaudin E, rebaudin F, rebaudin I, rebaudin H, rebaudin L, rebaudin K, rebaudin J, rebaudin M, rebaudin N, rebaudin O, durqueside A, durqueside B, stevia, stevioside, mogroside IV, mogroside V, mogroside VI, mogroside, cimenoside I, mogroside IIIE, mogaratin and its salts (monatin SS, RR, RS, SR), curculin, glycyrrhizic acid and its salts, sematrandrine, monellin, mabinlin, brazzein, hernandulcin, lecithin, phloroglucinol, root bark glycosides, root bark Glycosides, trifolioside, baiyunoside, osladin, polypodoside A, pterocaryoside A, pterocaryoside B, mukurozioside, phlomisoside I, periandrin I, abrusoside A, stevioside and cyclocaryaside I, sugar alcohols such as erythritol, sucralose, acetylsupanitate, acesulfame potassium and its salts, aspartame, alitane, saccharin and its salts, neohesperidin dihydrochalcone, cyclohexylsulfamic acid, cyclohexylsulfamic acid and its salts, neotame, saccharin, glycosylated steviol glycosides (GSG) and combinations thereof.

[0169] In one specific embodiment, the sweetener is at least one carbohydrate sweetener that provides calories.

[0170] In one embodiment, the sweetener is a caloric sweetener or a mixture of caloric sweeteners. In another embodiment, the caloric sweetener is selected from sucrose, fructose, glucose, high-fructose corn / starch syrup, beet sugar, cane sugar, and combinations thereof.

[0171] In another embodiment, the sweetener is a rare sugar selected from: allusion, sorbitol, lysose, ribulose, xylose, xylulose, D-allose, L-ribose, D-tagatose, L-glucose, L-fucose, L-arabinose, malinosose, kosherose, and combinations thereof.

[0172] In one embodiment, the sweetener composition comprises at least one additional mogroside. Mogrosides may be provided in pure form or as part of a mixture (i.e., a mogroside blend). Exemplary mogrosides include, but are not limited to, any of the following: grosmogroside I, mogroside IA, mogroside IE, 11-oxomogroside IA, mogroside II, mogroside IIA, mogroside IIB, mogroside IIE, 7-oxomogroside IIE, mogroside III, mogroside IIIE, 11-oxomogroside IIIE, 11-deoxymogroside III, mogroside IV, mogroside IVA, 11-oxomogroside IV, 11-oxomogroside IVA, mogroside V, Isomogroside V, 11-oxomonoside V, 11-deoxymonoside V, 7-oxomonoside V, 11-oxomonoside V, isomogroside V, mogroside VI, mogroside alcohol, 11-oxomonoside alcohol, mogroside I, 11-oxomonoside I, and isomers of mogroside I (e.g., those disclosed in 20170119032; the entire contents of that patent are incorporated herein by reference), particularly the 1,6-α isomer of mogroside I and combinations thereof. Further exemplary mogrosides include those described in U.S. Patent Application Publication 2016039864, the contents of which are incorporated herein by reference.

[0173] In one embodiment, the at least one additional substance is a sweetening agent, symmenoside I.

[0174] In another embodiment, the at least one additional substance is the 1,6-α isomer of symbioside I for sweetening (mogroside-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranoside-(1→2)]-[α-D-glucopyranoside-(1→6)]-β-D-glucopyranoside}).

[0175] In yet another embodiment, the at least one additional substance is mogroside V, which enhances sweetness.

[0176] In yet another embodiment, the at least one additional substance is a sweetening agent called leboside M.

[0177] In yet another embodiment, the at least one additional substance is a sweetening agent, leboside A.

[0178] In another embodiment, the at least one additional substance is a sweetening agent, leboside D.

[0179] IV. Flavor-enhancing compositions

[0180] In some embodiments, the mogrosides of the present invention are flavor enhancers. For example, in some embodiments, at least one mogroside of the present invention modulates one or more flavor properties of a consumer product sweetened with a non-sucrose sweetener, thereby making the consumer product taste more like a sucrose-sweetened consumer product. Exemplary flavor property modulation includes increasing sweetness, reducing or eliminating bitterness, reducing or eliminating bitterness retention, reducing or eliminating sourness, reducing or eliminating astringency, reducing or eliminating saltiness, reducing or eliminating metallic notes, improving mouthfeel, reducing or eliminating sweetness retention, and increasing sweetness onset. Multiple flavor properties of the sweetener can be modulated simultaneously, so that the consumer product as a whole has more characteristics of sucrose sweetening. Methods for quantifying the improvement of sucrose sweetening characteristics are known in the art and include, for example, taste testing and histogram plotting.

[0181] In one specific embodiment, the mogroside of the present invention is a sweetness enhancer or modifier. As used herein, the term "sweetness enhancer" refers to a compound that enhances, amplifies, or strengthens the perceived sweetness of a consumer product (e.g., a beverage) when the compound is present in the product at a concentration equal to or below the sweetness recognition threshold of the compound (i.e., a concentration at which the compound does not contribute any noticeable sweetness in the absence of one or more other sweeteners).

[0182] As used herein, the term "sweetness modifier" refers to a compound that, when present in a consumer product (e.g., a beverage) at a concentration equal to or below the sweetness recognition threshold of the compound, alters the taste characteristics (such as persistence, off-flavor, etc.) of the sweetness of the product.

[0183] The term "sweetness enhancer" is synonymous with the terms "sweetness synergist," "sweetness enhancer," "sweetness amplifier," and "sweetness fortifier."

[0184] In one embodiment, the mogroside of the present invention can be directly added to a consumer product, i.e., provided in the form of a compound rather than a composition, to enhance sweetness. In the aforementioned embodiment, the mogroside of the present invention is added to the consumer product at a concentration equal to or below its sweetness recognition threshold concentration, i.e., a sweetness enhancer. In a specific embodiment, the mogroside of the present invention is added to the consumer product at a concentration below its sweetness recognition threshold concentration, i.e., a sweetness enhancer.

[0185] In some embodiments, the mogroside of the present invention is a sweetness enhancer or modifier, and is added to a consumer product in an amount providing a mogroside concentration 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 lower than its sweetness recognition threshold.

[0186] In some embodiments, when compared to the sucrose equivalent (SE) of a consumer product in which the mogroside of the present invention is absent, the mogroside of the present invention enhances the SE of the consumer product 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%.

[0187] In other embodiments, at least one mogroside of the present invention can be added to consumer products in the form of a sweetness-enhancing composition. As used herein, the term "sweetness-enhancing composition" refers to the composition of the present invention as described above, wherein when the mogroside of the present invention is present in a sweetness-enhancing composition at an amount that provides a mogroside concentration equal to or below its sweetness recognition threshold, the composition enhances, amplifies, or strengthens the perceived sweetness of the consumer product (e.g., a beverage). In one specific embodiment, the mogroside of the present invention is present at an amount that provides a mogroside concentration below its sweetness recognition threshold.

[0188] Considering that the sweetness-enhancing composition may contain one or more sweetness enhancers or modifiers in addition to at least one mogroside of the present invention, in one embodiment, the sweetness-enhancing composition may contain one additional sweetness enhancer. In other embodiments, the composition may contain two or more additional sweetness enhancers. In embodiments using two or more sweetness enhancers or modifiers, each should be present at a concentration equal to or lower than its corresponding sweetness recognition threshold concentration.

[0189] The one or more other sweetness enhancers or modifiers mentioned are selected from, but not limited to, the group consisting of: 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-hesperidin dihydrochalcone, MG isomogrosaide V, 4-hydroxycinnamic acid, 4-methyl Oxycinnamic acid, 1-(2-hydroxyphenyl)-3-(4-pyridyl)-1-propanone, 4-ethoxybenzyl nitrile, 2-methoxy-5-(phenoxymethyl)-phenol, 1-(2,4-dihydroxyphenyl)-2-(3-methoxy-4-hydroxyphenyl)-ethyl ketone, hesperidin dihydrochalcone-4'-β-D-glucoside, hesperidin, 2,3',6-trihydroxy-4'-methoxydihydrochalcone, N-(3'-methoxy-4'-hydroxybenzyl)-2,4,6-trihydroxybenzamide, 3'-7-dihydroxy-4'-methoxyflavone, phlorizin, FEMA 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, Hesperidin dihydrochalcone, FEMA GRAS Flavor 4872, FEMA GRAS Flavor 4899, 4-amino-5-(cyclohexyloxy)-2-methylquinoline-3-carboxylic acid, Rebaudin M, Rebaudin N, Rebaudin O, Rebaudin C and combinations thereof.

[0190] In another specific embodiment, the mogroside of the present invention is a flavor enhancer. As used herein, the term "flavor enhancer" refers to a compound that, when present in a consumer product (e.g., a beverage) at a concentration equal to or below the compound's flavor recognition threshold (i.e., a concentration at which the compound does not contribute any noticeable flavor in the absence of any flavor components), enhances, amplifies, or strengthens the perception of flavor components (i.e., any substance that provides sweetness, sourness, saltiness, aroma, bitterness, metallic taste, etc.). The term "flavor recognition threshold," as commonly used herein, is the lowest known concentration of a compound that can be perceived by human taste as a particular flavor. Flavor recognition threshold concentrations are specific to a particular compound and can vary based on temperature, matrix, composition, and / or flavor system.

[0191] The term "flavor enhancer" is synonymous with the terms "flavor synergist," "flavor amplifier," and "flavor fortifier."

[0192] In one embodiment, at least one mogroside of the present invention is added directly to a consumer product, i.e., not as a composition, but as a compound to enhance flavor. In this embodiment, the mogroside of the present invention is added to the consumer product at a concentration equal to or below its flavor recognition threshold concentration, i.e., a flavor enhancer. In a specific embodiment, the mogroside of the present invention is added to the consumer product at a concentration below its flavor recognition threshold concentration, i.e., a flavor enhancer.

[0193] When compared with the flavor of a consumer product in which the mogroside of the present invention is absent, the mogroside of the present invention enhances the flavor of the consumer product 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%.

[0194] In other embodiments, at least one mogroside of the present invention can be added to consumer products in the form of a flavor-enhancing composition. As used herein, the term "flavor-enhancing composition" 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 wherein when the at least one mogroside of the present invention is present in the flavor-enhancing composition in an amount that provides a mogroside concentration equal to or below its flavor recognition threshold, the composition enhances, amplifies, or strengthens the perception of the flavor components of the consumer product when added to a consumer product (e.g., a beverage).

[0195] Compared to the detected flavor of the same ingredient in a consumer product without a flavor enhancer, the addition of a flavor-enhancing composition increases the detected flavor of at least one flavor component in the consumer product. Without being bound by theory, the flavor-enhancing composition is unlikely to provide any noticeable flavor to the consumer product to which it is added, because the flavor enhancer is present in the consumer product at a concentration equal to or below its flavor recognition threshold.

[0196] Suitable flavoring ingredients include, but are not limited to, vanillin, vanilla extract, mango extract, cinnamon, citrus, coconut, ginger, terpineol, almond, menthol (including menthol without 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 substances that impart flavor and may include natural or non-natural (synthetic) substances that are safe for human or animal use within generally acceptable limits. Non-limiting examples of proprietary flavoring agents include... Natural flavoring sweetener K14323 ( Darmstadt, Germany; Symrise TMNatural sweetness masking agents 161453 and 164126 (Symrise) TM Holzminden, Germany; NaturalAdvantage TM Bitterness inhibitors 1, 2, 9 and 10 (Natural Advantage) TM Freehold, New Jersey, USA and Sucramask TM (Creative Research Management, Stockton, California, USA)

[0197] In another embodiment, flavor-enhancing compositions comprising at least one mogroside of the present invention enhance flavor (either individually or as a whole) when added to consumer products. These flavoring agents include, but are not limited to, fruit flavoring agents (including tropical fruit flavoring agents) and vanilla-caramel flavoring agents.

[0198] V. Additives

[0199] On the other hand, the compositions described herein (i.e., the compositions described above and the consumer products described below) may contain one or more additional additives and / or functional ingredients.

[0200] 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.

[0201] In one embodiment, the composition further comprises one or more polyols. As used herein, the term "polyol" refers to a molecule containing more than one hydroxyl group. Polyols can be diols, triols, or tetraols containing 2, 3, and 4 hydroxyl groups, respectively. Polyols can also contain more than 4 hydroxyl groups, such as pentaols, hexaols, heptaols, etc., containing 5, 6, or 7 hydroxyl groups, respectively. Additionally, polyols can also be sugar alcohols, polyhydroxy alcohols, or polyols as a reduced form of carbohydrate, wherein the carbonyl group (aldehyde or ketone, reducing sugar) has been reduced to a primary or secondary hydroxyl group.

[0202] Non-limiting examples of polyols in some embodiments include maltitol, mannitol, sorbitol, lactitol, xylitol, isomaltitol, propylene glycol, glycerol, threitol, galactitol, palaginose, reduced isomaltooligosaccharide, reduced xylooligosaccharide, reduced gentianooligosaccharide, reduced maltose syrup, reduced glucose syrup, and sugar alcohols or any other carbohydrate that can be reduced without adversely affecting the taste.

[0203] 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, GABA (α-isomer, β-isomer, and / or δ-isomer), glutamine, hydroxyproline, taurine, valine, sarcosine, and their salts such as sodium or potassium salts or acid salts. Amino acid additives can also be in D- or L-configuration and in mono-, di-, or ternary forms of the same or different amino acids. Additionally, if appropriate, the amino acids can be α-, β-, γ-, and / or δ-isomers. In some embodiments, combinations of the above amino acids and their corresponding salts (e.g., their sodium, potassium, calcium, magnesium, or other alkali metal or alkaline earth metal salts, or acid salts) are also suitable additives. Amino acids can be natural or synthetic. Amino acids can also be modified. A modified amino acid is any amino acid in which at least one atom has been added, removed, substituted, or a combination thereof (e.g., N-alkyl amino acids, N-acyl amino acids, or N-methyl amino acids). Non-limiting examples of modified amino acids include amino acid derivatives such as trimethylglycine, N-methylglycine, and N-methylalanine. 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 monosodium glutamate). Polyamino acid additives may also be D- or L-configurations. Additionally, if appropriate, polyamino acids may be α-, β-, γ-, δ-, and ε-isomers. In some embodiments, combinations of the above polyamino acids and their corresponding salts (e.g., their sodium, potassium, calcium, magnesium, or other alkali metal or alkaline earth metal salts or acid salts) are also suitable additives. The polyamino acids described herein may also include copolymers of different amino acids. Polyamino acids may be natural or synthetic. Polyamino acids can also be modified by adding, removing, substituting, or combining at least one atom (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 with different molecular weights (MW), such as poly-L-α-lysine having MW of 1,500, 6,000, 25,200, 63,000, 83,000, or 300,000.

[0204] Suitable sugar acid additives include, but are not limited to, aldonic acid, glucuronic acid, aldonic acid, alginic acid, gluconic acid, glucuronic acid, gluconic acid, galactonic acid, galacturonic acid, and their salts (e.g., sodium salts, potassium salts, calcium salts, magnesium salts or other physiologically acceptable salts) and combinations thereof.

[0205] 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, their alkali metal salts or alkaline earth metal salts, and combinations thereof. The nucleotides described herein may also contain nucleotide-related additives, such as nucleosides or nucleic acid bases (e.g., guanine, cytosine, adenine, thymine, uracil).

[0206] Suitable organic acid additives include any compound containing a -COOH moiety, such as C2-C30 carboxylic acids, substituted hydroxy C2-C30 carboxylic acids, butyric acid (ethyl ester), substituted butyric acid (ethyl ester), benzoic acid, substituted benzoic acid (e.g., 2,4-dihydroxybenzoic acid), substituted cinnamic acid, hydroxy acids, substituted hydroxybenzoic acid, anisic acid, substituted cyclohexyl carboxylic acids, tannic acid, aconitic acid, lactic acid, tartaric acid, citric acid, isocitric acid, gluconic acid, glucoheponic acid, adipic acid, hydroxycitric acid, malic acid, fruitaric acid (a blend of malic acid, fumaric acid, and tartaric acid), fumaric acid, maleic acid, succinic acid, chlorogenic acid, salicylic acid, creatine, caffeic acid, bile acids, acetic acid, ascorbic acid, alginic acid, isoascorbic acid, polyglutamic acid, gluconic acid δ-lactone, and their alkali metal salts or alkaline earth metal salt derivatives. In addition, organic acid additives can also be in D-configuration or L-configuration.

[0207] Suitable organic acid additive salts include, but are not limited to, sodium, calcium, potassium, and magnesium salts of all organic acids, such as citrate, malate, tartrate, fumarate, lactate (e.g., sodium lactate), alginate (e.g., sodium alginate), ascorbate (e.g., sodium ascorbate), benzoate (e.g., sodium benzoate or potassium benzoate), sorbate, and adipate. Examples of the organic acid additives may optionally be substituted with at least one group selected from the following: hydrogen, alkyl, alkenyl, alkynyl, halogen, haloalkyl, carboxyl, acyl, acyloxy, amino, amide, carboxyl derivative, alkylamino, dialkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonyl, thiol, imine, sulfonyl, alkylthio, sulfinyl, aminosulfonyl, carboxyalkoxy, formamide, phosphonyl, oxyphosphonyl, phosphoryl, phosphonyl, thioester, thioether, acid anhydride, oxime, hydrazyl, carbamoyl, phosphoro, or phosphonate.

[0208] 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 their alkali metal salts or alkaline earth metal salts (e.g., inositol hexaphosphate Mg / Ca).

[0209] Suitable bitter compound additives include, but are not limited to, caffeine, quinine, urea, bitter orange oil, naringin, quassula and its salts.

[0210] 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 menthol without 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 substances that impart flavor and may include natural or non-natural (synthetic) substances that are safe for human or animal use within generally acceptable limits. Non-limiting examples of proprietary flavoring agents include... Natural flavoring sweetener K14323 ( Darmstadt, Germany; Symrise TM Natural sweetness masking agents 161453 and 164126 (Symrise) TM Holzminden, Germany; Natural Advantage TM Bitterness inhibitors 1, 2, 9 and 10 (Natural Advantage) TM Freehold, New Jersey, USA and SucramaskTM (Creative Research Management, Stockton, California, USA)

[0211] Suitable polymer additives include, but are not limited to, chitosan, pectin, pectic acid, pectinuric acid, polyuronic acid, polygalacturonic acid, starch, food hydrolysates or their crude extracts (e.g., Senegalese gum arabic (Fibergum arabic)). TM Seyal gum arabic, 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, alginate, sodium alginate, propylene glycol alginate, and polyethylene glycol sodium alginate, sodium hexametaphosphate and its salts, as well as other cationic and anionic polymers.

[0212] Suitable protein or protein hydrolysate additives include, but are not limited to, bovine serum albumin (BSA), whey protein (including its fractions or concentrates, 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 isolates, protein hydrolysates, reaction products of protein hydrolysates, glycoproteins and / or proteoglycans containing amino acids (e.g. glycine, alanine, serine, threonine, asparagine, glutamine, arginine, valine, isoleucine, leucine, n-valine, methionine, proline, tyrosine, hydroxyproline, etc.), collagen (e.g., gelatin), partially hydrolyzed collagen (e.g., hydrolyzed fish collagen), and collagen hydrolysates (e.g., porcine collagen hydrolysates).

[0213] Suitable surfactant additives include, but are not limited to, polysorbates (e.g., polyoxyethylene sorbitan monooleate (polysorbate 80), polysorbate 20, polysorbate 60), sodium dodecylbenzene sulfonate, dioctyl sulfosuccinate or sodium dioctyl sulfosuccinate, sodium dodecyl sulfate, hexadecylpyridinium chloride (hexadecylpyridinium chloride), hexadecyltrimethylammonium bromide, sodium cholate, carbamoyl, choline chloride, sodium glycocholate, sodium taurodeoxycholate, lauroyl arginine ester, sodium stearoyl lactylate, sodium taurocholate, lecithin, sucrose oleate, sucrose stearate, sucrose palmitate, sucrose laurate, and other emulsifiers.

[0214] Suitable flavonoid additives are classified as flavonols, flavones, flavanones, flavan-3-ols, isoflavones, or anthocyanins. Non-limiting examples of flavonoid additives include, but are not limited to, catechins (e.g., green tea extracts, such as polyphenonium). TM 60. Polyphenon TM 30 and Polyphenon TM 25 (Mitsui Norin Co., Ltd., Japan), polyphenols, rutin (e.g., enzyme-modified rutin Sanmelin) TM AO (San-fi Gen FFI, Inc., Osaka, Japan), neohesperidin, naringin, neohesperidin dihydrochalcone, etc.

[0215] Suitable alcohol additives include, but are not limited to, ethanol.

[0216] Suitable astringent compound additives include, but are not limited to, tannic acid, europium chloride (EuCl3), gadolinium chloride (GdCl3), terbium chloride (TbCl3), alum, tannic acid, and polyphenols (e.g., tea polyphenols).

[0217] The compositions provided herein may also contain one or more functional ingredients that provide actual or perceived health benefits to the composition. Functional ingredients include, but are not limited to, saponins, antioxidants, dietary fiber sources, fatty acids, vitamins, glucosamine, minerals, preservatives, hydrating agents, probiotics, prebiotics, post-biotics, weight management agents, osteoporosis management agents, phytoestrogens, long-chain primary aliphatic saturated alcohols, phytosterols, and combinations thereof.

[0218] Exemplary functional ingredients include, but are not limited to, saponins, antioxidants, dietary fiber sources, fatty acids, vitamins, glucosamine, minerals, preservatives, hydrating agents, probiotics, prebiotics, weight management agents, osteoporosis management agents, phytoestrogens, long-chain aliphatic saturated primary alcohols, phytosterols, and combinations thereof.

[0219] In some embodiments, the functional ingredient is at least one saponin. As used herein, the at least one saponin may comprise a single saponin or multiple saponins as functional ingredients of the compositions provided herein. Saponins are natural plant products containing an aglycone ring structure and one or more sugar moieties. Non-limiting examples of specific saponins used in specific embodiments of the invention include group A acetyl saponins, group B acetyl saponins, and group E acetyl saponins. Several common sources of saponins include soybeans, soapwort plants (of the genus *Saponaria*, whose roots have historically been used as soap), as well as alfalfa, aloe vera, asparagus, grapes, chickpeas, yucca, and various other legumes and wild herbs, having a saponin content of approximately 5% by weight on a dry basis. 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 U.S. Patent Application No. 2005 / 0123662, the disclosure of which is expressly incorporated herein by reference.

[0220] In some embodiments, the functional ingredient is at least one antioxidant. As used herein, "antioxidant" means any substance that inhibits, suppresses, or reduces oxidative damage to cells and biomolecules. Examples of suitable antioxidants for use in embodiments of the invention include, but are not limited to, vitamins, vitamin cofactors, minerals, hormones, carotenoids, carotenoid terpenes, non-carotenoid terpenes, flavonoids, flavonoid polyphenols (e.g., bioflavonoids), flavonols, flavonoids, phenols, polyphenols, phenolic esters, polyphenolic esters, non-flavonoid phenols, isothiocyanates, and combinations thereof. In some embodiments, the antioxidants are vitamin A, vitamin C, vitamin E, ubiquinone, minerals selenium and manganese, melatonin, α-carotene, β-carotene, lycopene, lutein, zeanthin, cryptoxanthin, resveratrol, eugenol, quercetin, catechin, gossypol, hesperidin, curcumin, ferulic acid, thymol, hydroxytyrosol, turmeric, thyme, olive oil, alpha-lipoic acid, glutathione, glutamine, oxalic acid, tocopherol derivatives, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediaminetetraacetic acid (EDTA), tert-butylhydroquinone, acetic acid, pectin, tocotrienols, tocopherol, coenzyme Q10, zeaxanthin, astaxanthin, canthaxanthin, and saponins. Limonene, kaempfedrol, myricetin, isorhamnetin, proanthocyanidins, quercetin, rutin, luteolin, apigenin, tangeritin, hesperidin, naringenin, erodictyol, flavan-3-ols (e.g., anthocyanins), gallocatechin, epicatechin and its gallate esters, epigallocatechin and its gallate esters (ECGC), theaflavins and their gallate esters, thearubigins, isoflavones, phytoestrogens, genistein, daidzein, stigmacanoidin, anythocyanin, cyaniding, delphinidin, malvidin, malvidin, methyl anthocyanin, petunidin, ellagic acid, gallic acid, salicylic acid, rosmarinic acid, cinnamic acid and its derivatives (e.g., ferulic acid), chlorogenic acid, chicoric acid (acid), gallnut tannins, ellagic acid, β-anthocyanin and other plant pigments, silymarin, citric acid, lignans, antinutrients, bilirubin, uric acid, R-α-lipoic acid, N-acetylcysteine, emblicanin, apple extract, apple peel extract (apple polyphenols), red rooibos extract, green rooibos extract,Green), hawthorn fruit extract, raspberry extract, green coffee antioxidant (GCA), 20% wild cherries extract, grape seed extract (VinOseed), cocoa bean extract, hops extract, mangosteen fruit extract, mangosteen shell extract, cranberry extract, pomegranate extract, pomegranate peel extract, pomegranate seed extract, hawthorn berry extract, pomella pomegranate extract, cinnamon bark extract, grape skin extract, blueberry extract, pine bark extract, pycnogenol, elderberry extract, mulberry root extract, goji berry extract, blackberry extract, blueberry extract, blueberry leaf extract, raspberry extract, turmeric extract, citrus bioflavonoids, blackcurrant, ginger, acai berry powder, green coffee bean extract, green tea extract, and phytic acid or combinations thereof. In alternative embodiments, the antioxidant is a synthetic antioxidant, such as butylated hydroxytoluene or butylated hydroxyanisole. Other sources of suitable antioxidants for use in embodiments of the present invention include, but are not limited to, fruits, vegetables, tea, cocoa, chocolate, spices, herbs, rice, organ meats from livestock, yeast, whole grains, or cereal grains.

[0221] Specific antioxidants belong to a class of phytonutrients called polyphenols (also known as "polyphenols"), which are a group of chemical substances found in plants, characterized by the presence of more than one phenolic group per molecule. A variety of health benefits can be derived from polyphenols, including, for example, prevention of cancer, heart disease, and chronic inflammatory diseases, as well as improvement of mental and physical energy. Suitable polyphenols used in embodiments of the present invention include catechins, proanthocyanidins, proanthocyanidins, anthocyanins, quercetin, rutin, resveratrol, isoflavones, curcumin, punicin, ellagitannins, hesperidin, naringin, citrus flavonoids, chlorogenic acid, other similar materials, and combinations thereof.

[0222] In a specific embodiment, the antioxidant is a catechin, such as epigallocatechin gallate (EGCG). In another embodiment, the antioxidant is selected from proanthocyanidins, proanthocyanidins, or combinations thereof. In a specific embodiment, the antioxidant is anthocyanin. In still other embodiments, the antioxidant is selected from quercetin, rutin, or combinations thereof. In other embodiments, the antioxidant is resveratrol. In other embodiments, the antioxidant is isoflavone. In other embodiments, the antioxidant is curcumin. In other embodiments, the antioxidant is selected from quercetin, ellagitannins, or combinations thereof. In other embodiments, the antioxidant is chlorogenic acid.

[0223] In some embodiments, the functional ingredient is at least one source of dietary fiber. A variety of polymeric carbohydrates with significantly different structures in both composition and bonding 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-glucan, pectin, gums, mucilage, waxes, inulin, oligosaccharides, fructooligosaccharides, cyclodextrin, chitin, and combinations thereof. Although dietary fiber is generally derived from plant sources, recalcitrant animal products such as chitin are also classified as dietary fiber. Chitin is a polysaccharide composed of acetylglucosamine units linked by β(1-4) bonds similar to those in cellulose.

[0224] In some embodiments, the functional ingredient is at least one fatty acid. As used herein, "fatty acid" refers to any straight-chain monocarboxylic acid and includes saturated fatty acids, unsaturated fatty acids, long-chain fatty acids, medium-chain fatty acids, short-chain fatty acids, fatty acid precursors (including ω-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 having a long aliphatic tail. As used herein, "ω-3 fatty acid" refers to any polyunsaturated fatty acid having a first double bond as the third carbon-carbon bond from the terminal methyl end of its carbon chain. In specific embodiments, ω-3 fatty acids may include long-chain ω-3 fatty acids. As used herein, "ω-6 fatty acid" refers to any polyunsaturated fatty acid having a first double bond as the sixth carbon-carbon bond from the terminal methyl end of its carbon chain.

[0225] Suitable ω-3 fatty acids used in embodiments of the present invention may be derived from, for example, algae, fish, animals, plants, or combinations thereof. Examples of suitable ω-3 fatty acids include, but are not limited to, linolenic acid, alpha-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, octadecanoic acid, eicosapentaenoic acid, and combinations thereof. In some embodiments, suitable ω-3 fatty acids may be provided in fish oil (e.g., herring oil, tuna oil, salmon oil, bonito oil, and cod oil), microalgae ω-3 oil, or combinations thereof. In specific embodiments, suitable omega-3 fatty acids can 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) (from Governor Naho, Illinois)); bonito oil and MEG-3 (from Ocean Nutrition, Dartmouth, NS) (from Symrise, Holzminden, Germany) (from Symrise, Holzminden, Germany) (from tuna or salmon) Wilton, CT), OmegaSource2000, marine oil from herring and marine oil from cod (from OmegaSource, RTP, NC, Research Triangle, North Carolina).

[0226] Suitable ω-6 fatty acids include, but are not limited to, linoleic acid, gamma-linolenic acid, dihomeno-gamma-linolenic acid, arachidonic acid, eicosadienoic acid, docosadienoic acid, adrenaline, docosapentaenoic acid, and combinations thereof.

[0227] Suitable esterified fatty acids used in embodiments of the present invention may include, but are not limited to, monoacylglycerols containing ω-3 and / or ω-6 fatty acids, diacylglycerols containing ω-3 and / or ω-6 fatty acids, or triacylglycerols containing ω-3 and / or ω-6 fatty acids, and combinations thereof.

[0228] In some 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.

[0229] Many other compounds have been officially classified as vitamins. These compounds may be referred to as pseudovitamins and include, but are not limited to, compounds such as ubiquinone (coenzyme Q10), panthenine, dimethylglycine, taestrile, amygdalin, flavonoids, para-aminobenzoic acid, adenine, adenosine, and S-methylmethionine. As used herein, the term vitamin includes pseudovitamins. In some embodiments, vitamins are selected from fat-soluble vitamins such as vitamin A, vitamin D, vitamin E, vitamin K, and combinations thereof. In other embodiments, vitamins are selected from water-soluble vitamins such as vitamin B1, vitamin B2, vitamin B3, vitamin B6, vitamin B12, folic acid, biotin, pantothenic acid, vitamin C, and combinations thereof.

[0230] In some embodiments, the functional ingredient is glucosamine, optionally further comprising chondroitin sulfate.

[0231] In some embodiments, the functional ingredient is at least one mineral. According to the teachings of the present invention, minerals comprise inorganic chemical elements necessary for living organisms. Minerals consist of a wide range of compositions (e.g., elements, simple salts, and complex silicates) and have widely different crystal structures. They may occur naturally in foods and beverages, may be added as supplements, or may be consumed or administered separately from food or beverages.

[0232] Minerals can be classified as bulk minerals, which are required in relatively large quantities, or trace minerals, which are required in relatively small quantities. Bulk minerals are typically required in amounts greater than or equal to about 100 mg / day, while trace minerals are those required in amounts less than about 100 mg / day.

[0233] In one embodiment, the mineral is selected from major minerals, trace minerals, or combinations thereof. Non-limiting examples of major minerals include calcium, chlorine, magnesium, phosphorus, potassium, sodium, and sulfur. Non-limiting examples of trace minerals include chromium, cobalt, copper, fluorine, iron, manganese, molybdenum, selenium, zinc, and iodine. Although iodine is generally classified as a trace mineral, it requires a larger quantity than other trace minerals and is often classified as a major mineral.

[0234] In one specific embodiment, the mineral is a trace mineral that is considered essential for human nutrition, and non-limiting examples include bismuth, boron, lithium, nickel, rubidium, silicon, strontium, tellurium, tin, titanium, tungsten, and vanadium.

[0235] The minerals presented herein may be in any form known to those skilled in the art. For example, in one specific embodiment, the mineral may be in its ionic form, having a positive or negative charge. In another specific embodiment, the mineral may be in its molecular form. For example, sulfur and phosphorus typically exist naturally as sulfates, sulfides, and phosphates.

[0236] In some embodiments, the functional ingredient is at least one preservative. In specific embodiments of the invention, the preservative is selected from antimicrobial agents, antioxidants, antienzymes, 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 another embodiment, the preservative is a sorbate. Sorbic acid includes, but is not limited to, potassium sorbate, sodium sorbate, calcium sorbate, and sorbic acid. In yet another embodiment, the preservative is a nitrate and / or a nitrite. Nitrates and nitrites include, but are not limited to, sodium nitrate and sodium nitrite. In another embodiment, at least one preservative is a bacteriocin, such as nifedipine. In another embodiment, the preservative is ethanol. In yet another embodiment, the preservative is ozone. Non-limiting examples of enzyme-resistant agents suitable for use as preservatives in specific embodiments of the invention include ascorbic acid, citric acid, and metal chelating agents such as ethylenediaminetetraacetic acid (EDTA).

[0237] In some embodiments, the functional ingredient is at least one hydrating agent. In specific embodiments, the hydrating agent is an electrolyte. Non-limiting examples of electrolytes include sodium, potassium, calcium, magnesium, chlorides, phosphates, bicarbonates, and combinations thereof. Suitable electrolytes for specific embodiments of the invention are also described in U.S. Patent No. 5,681,569, the disclosure of which is expressly incorporated herein by reference. In one embodiment, the electrolyte is obtained from its corresponding water-soluble salt. Non-limiting examples of salts for specific embodiments include chlorides, carbonates, sulfates, acetates, bicarbonates, citrates, phosphates, hydrogen phosphates, tartrates, sorbates, citrates, benzoates, or combinations thereof. In other embodiments, the electrolyte is provided by juicing, fruit extracts, vegetable extracts, tea, or tea extracts.

[0238] In a specific embodiment of the invention, the hydrating agent is a carbohydrate that replenishes the energy stored by muscles. Suitable carbohydrates for use in specific embodiments of the 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 specific embodiments include trioses, tetroses, pentoses, hexoses, heptoses, octoses, and nonoses. Non-limiting examples of suitable monosaccharides of specific types include glyceraldehyde, dihydroxyacetone, erythrose, thoraxose, erythritolose, arabinose, lysose, ribose, xylose, ribulose, xylulose, allose, azoose, galactose, glucose, gulose, idulose, mannose, talose, fructose, allulose, sorbose, tagatose, mannoheptulose, sedoheltulose, octolose, and sialose. 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, carbohydrates are provided via corn syrup, beet sugar, cane sugar, juice, or tea.

[0239] In another specific embodiment, the hydrating agent is a flavanol that provides cell rehydration. Flavanols are a class of naturally occurring substances found in plants and typically comprise a 2-phenylbenzopyranone molecular skeleton attached to one or more chemical moieties. Non-limiting examples of suitable flavanols used in specific embodiments of the invention include catechins, epicatechin, gallocatechin, epigallocatechin, epicatechin gallate, epigallocatechin 3-galate, theaflavins, theaflavins 3-galate, theaflavins 3'-galate, theaflavins 3,3'-galate, thearubigins, or combinations thereof. Several common sources of flavanols include tea plants, fruits, vegetables, and flowers. In a preferred embodiment, flavanols are extracted from green tea.

[0240] In one specific embodiment, the hydrating agent is a glycerol solution that enhances exercise endurance. Ingestion of glycerol-containing solutions has been shown to provide a variety of beneficial physiological effects, such as increased blood volume, reduced heart rate, and decreased rectal temperature.

[0241] In some embodiments, the functional ingredient is selected from at least one probiotic, prebiotic, or combination thereof. Probiotics are beneficial microorganisms that influence the naturally occurring gastrointestinal microbiota of the human body. Examples of probiotics include, but are not limited to, bacteria of the genera *Lactobacillus*, *Bifidobacteria*, *Streptococcus*, or combinations thereof that provide beneficial effects to humans. In a specific embodiment of the invention, the at least one probiotic is selected from the genus *Lactobacillus*. According to other specific embodiments of the invention, the probiotic is selected from the genus *Bifidobacteria*. According to other specific embodiments of the invention, the probiotic is selected from the genus *Streptococcus*.

[0242] The probiotics that can be used according to the present invention are well known to those skilled in the art. Non-limiting examples of foods containing probiotics include yogurt, German sauerkraut, kefir, Korean kimchi, fermented vegetables, and other foods containing microbial elements that beneficially influence the host animal by improving intestinal microbial homeostasis.

[0243] According to the teachings of this invention, prebiotics include, but are not limited to, mucopolysaccharides, oligosaccharides, polysaccharides, amino acids, vitamins, nutrient precursors, proteins, and combinations thereof. In specific embodiments of this invention, prebiotics are selected from dietary fiber, including, but not limited to, polysaccharides and oligosaccharides. Non-limiting examples of oligosaccharides classified as prebiotics according to specific embodiments of this invention include fructooligosaccharides, inulin, isomaltooligosaccharides, lactitol, lactulose oligosaccharides, lactulose, dextrin, soybean oligosaccharides, trans-galactose-oligosaccharides, and xylooligosaccharides. In other embodiments, prebiotics are amino acids. Although many known prebiotics break down to provide carbohydrates for probiotics, some probiotics also require amino acids to provide nutrients.

[0244] Prebiotics are naturally found in a variety of foods, including but not limited to bananas, berries, asparagus, garlic, wheat, oats, barley (and other whole grains), flaxseed, tomatoes, Jerusalem artichoke, onions and chicory, leafy greens (e.g., dandelion leaves, spinach, kale leaves, beets, headless kale, mustard greens, turnip leaves), and legumes (e.g., lentils, kidney beans, chickpeas, navy beans, white beans, black beans).

[0245] In some embodiments, the functional ingredient is at least one weight management agent.

[0246] As used herein, “weight management agents” include appetite suppressants and / or thermic agents. As used herein, the phrases “appetite suppressant,” “appetite-satisfying composition,” “satisfying agent,” and “satisfying ingredient” are synonymous. The phrase “appetite suppressant” describes a macronutrient, herbal extract, exogenous hormone, appetite suppressant, anorexia nervosa, drug, or combination thereof that, when delivered in an effective amount, suppresses, inhibits, reduces, or otherwise diminishes a person’s appetite. The phrase “thermic agent” describes a macronutrient, herbal extract, exogenous hormone, appetite suppressant, anorexia nervosa, drug, or combination thereof that, when delivered in an effective amount, activates or otherwise enhances a person’s thermic action or metabolism.

[0247] Suitable weight management agents include macronutrients selected from groups of the following: protein, carbohydrates, dietary fat, and combinations thereof. The consumption of protein, carbohydrates, and dietary fat stimulates the release of peptides that suppress appetite. For example, the consumption of protein and dietary fat stimulates the release of the gastrointestinal hormone cholecystokinin (CCK), while the consumption of carbohydrates and dietary fat stimulates the release of glucagon-like peptide-1 (GLP-1).

[0248] Suitable macronutrient weight management agents also include carbohydrates. Carbohydrates generally include sugars, starches, cellulose, and gums that are converted into glucose by the body for energy. Carbohydrates are generally divided into two categories: digestible carbohydrates (e.g., monosaccharides, disaccharides, and starches) and indigestible carbohydrates (e.g., dietary fiber). Studies have shown that indigestible carbohydrates and complex polymeric carbohydrates with reduced absorption and digestibility in the small intestine stimulate physiological responses that inhibit food intake. Therefore, the carbohydrates presented herein ideally include indigestible carbohydrates or carbohydrates with reduced digestibility. Non-limiting examples of such carbohydrates include polydextrose; inulin; monosaccharide-derived polyols such as erythritol, mannitol, xylitol, and sorbitol; disaccharide-derived alcohols such as isomaltitol, lactitol, and maltitol; and hydrogenated starch hydrolysates. Carbohydrates are described in more detail below.

[0249] In another specific embodiment, the weight management agent is dietary fat. Dietary fat is a lipid comprising a combination of saturated and unsaturated fatty acids. Polyunsaturated fatty acids have been shown to have a greater satiating capacity than monounsaturated fatty acids. Therefore, the dietary fats presented herein ideally include polyunsaturated fatty acids, with triacylglycerols being a non-limiting example.

[0250] In one specific embodiment, the weight management agent is an herbal extract. Extracts from a variety of plant types have been identified as having appetite-suppressing properties. Non-limiting examples of plants whose extracts have appetite-suppressing 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, gum resin, and blackcurrant seed oil.

[0251] Herbal extracts can be prepared from any type of plant material or plant biomass. Non-limiting examples of plant material and biomass include stems, roots, leaves, dried powders obtained from plant material, and sap or dried sap. Herbal extracts are typically prepared by extracting sap from a plant and then spray-drying the sap. Alternatively, solvent extraction procedures can be used. After the initial extraction, it may be desirable to further fractionate the initial extract (e.g., by column chromatography) to obtain a herbal extract with enhanced activity. Such techniques are well known to those skilled in the art.

[0252] In one specific embodiment, the herbal extract is derived from plants of the genus *Hypericum*, including species such as *H. alstonii*, *H. curlorii*, *H. dregei*, *H. flava*, *H. gordonii*, *H. jutatae*, *H. mossamedensis*, *H. officinalis*, *H. parviflorai*, *H. pedicellata*, *H. pilifera*, *H. ruschii*, and *H. triebneri*. *Hypericum* plants are succulent plants native to South Africa. A sterol glycoside called P57 from the genus *Hypericum* is believed to be responsible for the appetite-suppressing effect of *Hypericum* species. In another specific embodiment, the herbal extract is derived from plants of the genus *Caralluma*, which includes species such as *C. indica*, *C. fimbriata*, *C. attenuate*, *C. tuberculata*, *C. edulis*, *C. adscendens*, *C. stalammifera*, *C. umbellate*, *C. penicillata*, *C. russeliana*, *C. retrospicens*, *C. Arabica*, and *C. lasiantha*. *Caralluma* plants belong to the same subfamily as the genus *Caralluma*, the family Asclepiadaceae. Buffalo paw is a small, upright, succulent plant native to India with medicinal properties such as appetite suppression. These medicinal properties are generally attributed to glycosides belonging to the glycoside pregnane group. Non-limiting examples of these glycosides 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 plants of the genus *Trichocaulon*. *Trichocaulon* plants are succulent plants typically native to South Africa, similar to the genus *Trichocaulon*, and include species *Trichocaulon piliferum* and *Trichocaulon officinale*.In another specific embodiment, the herbal extract is derived from plants of the genera *Stapelia* or *Orbea*, including species such as *S. gigantea* and *O. variegate*. Both *Stapelia* and *O. variegate* belong to the same subfamily as *Pyridae*, the Asclepias family. Without being bound by any theory, compounds believed to exhibit appetite-suppressing activity are saponins, such as pregnane glycosides, including stavaroside A, B, C, D, E, F, G, H, I, J, and K. In another specific embodiment, the herbal extract is derived from plants of the genus *Asclepias*. *Asclepias* plants also belong to the Asclepias family. Non-limiting examples of *Asclepias* plants include *A. incarnate*, *A. curassayica*, *A. syriaca*, and *A. tuberose*. Not wishing to be bound by any theory, the extract is believed to contain steroidal compounds with appetite-suppressing effects, such as pregnane glycosides and pregnane aglycones.

[0253] In one specific embodiment, the weight management agent is an exogenous hormone with weight management function. Non-limiting examples of such hormones include CCK, peptide YY, ghrelin, bufotenoid and gastrin-releasing peptide (GRP), enterostatin, apolipoprotein A-IV, GLP-1, amyloidin, somastatin, and leptin.

[0254] In another embodiment, the weight management agent is a drug. Non-limiting examples include phenbutylamine, diethylamine benzophenone, benzotriazine, sibutramine, rimonaban, gastrin, fluoxetine hydrochloride, ephedrine, phenethylamine, or other stimulants.

[0255] In some embodiments, the functional ingredient is at least one osteoporosis management agent. In some embodiments, the osteoporosis management agent is at least one calcium source. According to specific embodiments, the calcium source is any compound containing calcium, including calcium salt complexes, dissolved substances, and other forms. 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, dissolved substances thereof, and combinations thereof.

[0256] According to one specific embodiment, the osteoporosis management agent is a magnesium source. A magnesium source is any compound containing magnesium, including magnesium salt complexes, dissolved substances, and other forms. Non-limiting examples of magnesium sources include magnesium chloride, magnesium citrate, magnesium gluconate, magnesium gluconate, magnesium lactate, magnesium hydroxide, magnesium picolate, magnesium sulfate, dissolved substances thereof, and mixtures thereof. In another specific embodiment, the magnesium source comprises amino acid chelated magnesium or creatine chelated magnesium.

[0257] In other embodiments, the osteoporosis agent is selected from vitamins D, C, K, their precursors and / or β-carotene and combinations thereof.

[0258] Many plants and plant extracts have also been identified as effective in the prevention and treatment of osteoporosis. Non-limiting examples of suitable plants and plant extracts as osteoporosis management agents include species of the genera *Taraxacum* and *Amelanchier* as disclosed in U.S. Patent Publication No. 2005 / 0106215, and species of the genera *Lindera*, *Artemisia*, *Acorus*, *Carthamus*, *Carum*, *Cnidium*, *Curcuma*, *Cyperus*, and *Juniperus*. s), Prunus, Iris, Cichorium, Dodonaea, Epimedium, Erigorum, Soya, Mentha, Ocimum, thymus, Tanacetum, Plantago, Spearmint, Bixa, Vitis, Rosemarinus, Rhus, and Anethum.

[0259] In some embodiments, the functional ingredient is at least one phytoestrogen. Phytoestrogens are compounds found in plants that can typically be delivered to the human body by ingestion of plants or plant parts containing phytoestrogens. As used herein, "phytoestrogen" means any substance that causes any degree of estrogen-like effect when introduced into the body. For example, phytoestrogens can bind to estrogen receptors in the body and have a small estrogen-like effect. Examples of suitable phytoestrogens for use in embodiments of the invention include, but are not limited to, isoflavones, stilbenes, lignans, resorcyclic acidlactone, coumarins, coumestan, coumestrobin, equol, and combinations thereof. Suitable sources of phytoestrogens include, but are not limited to, whole grains, cereals, fiber, fruits, vegetables, black cohosh, agave root, blackcurrant, cherry pod, chasteberry, spasmodic bark, angelica root, devil's club root, false unicorn root, ginseng root, *Gnaphalium affine*, licorice juice, *Gnaphalium affine*, motherwort, peony root, raspberry leaves, Rosaceae plants, sage leaves, sage root, *Salvia splendens* seeds, wild yam root, flowering yarrow, legumes, soybeans, soybean products (e.g., miso, soy flour, soy milk, soy nuts, soy protein isolate, tempen, or tofu), chickpeas, nuts, lentils, seeds, clover, red clover, dandelion leaves, dandelion root, fenugreek seeds, green tea, hops, red wine, flaxseed, garlic, onion, flaxseed, borage, and butterfly tubers. Weed, caraway, privet tree, vitex, jujube, dill, fennel seeds, centella asiatica, milk thistle, mint, pomegranate, artemisia annua, bean flour, chrysanthemum, kudzu root, and combinations thereof.

[0260] Isoflavones belong to the group of phytonutrients called polyphenols. Generally, polyphenols (also known as "polyphenols") are a group of chemical substances found in plants, characterized by the presence of more than one phenolic group per molecule.

[0261] Suitable phytoestrogens and isoflavones according to embodiments of the present invention include genistein, daidzein, genistein, chickpea extract A, gentiocarbazone, their respective naturally occurring glycosides and glycoside conjugates, maltosine, open-ring isolar resinin, enteroester, enterodiol, plant tissue protein, and combinations thereof.

[0262] Suitable sources of isoflavones used in embodiments of the present invention include, but are not limited to, soybeans, soybean products, legumes, alfalfa sprouts, chickpeas, peanuts, and red clover.

[0263] In some embodiments, the functional ingredient is at least one long-chain aliphatic saturated primary alcohol. Long-chain aliphatic saturated primary alcohols are organic compounds from different groups. The term "alcohol" refers to the fact that these compounds are characterized by a hydroxyl group (-OH) bonded to a carbon atom. Non-limiting examples of specific long-chain aliphatic saturated primary alcohols used in specific embodiments of the invention include 1-octanol (C8), 1-nonanol (C9), 1-decanol (C10), 1-dodecanol (C12), 1-tetradecanol (C14), 1-hexadecanol (C16), 1-octadecanol (C18), 1-eicosanool (C20), 1-docodecanol (C22), 1-tetracosanool (C24), 1-hexadecanol (C26), 1-heptadecanol (C27), 1-octanosol (C28), 1-nonadecanol (C29), 1-triacontanol (C30), 1-triadecanol (C32), and 1-triacontanol (C34).

[0264] In a particularly desirable embodiment of the invention, the long-chain primary aliphatic saturated alcohol is a prilool. Prilool is a term relating to a mixture of long-chain aliphatic saturated primary alcohols consisting primarily of: 28-carbon 1-octacosanol and 30-carbon 1-triacontanol, and other alcohols in lower concentrations such as 22-carbon 1-docosahexacontanol, 24-carbon 1-tetracosanol, 26-carbon 1-hexacosanol, 27-carbon 1-heptacosanol, 29-carbon 1-nonacontanol, 32-carbon 1-trimosahexacontanol, and 34-carbon 1-trimosahexacontanol.

[0265] In some embodiments, the functional ingredient is at least one phytosterol, phytosterol, or a combination thereof. As used herein, the phrases “sterol,” “phytosterol,” and “plant sterol” are synonymous. Phytosterols and sterols are naturally found in small amounts in many fruits, vegetables, nuts, seeds, grains, legumes, vegetable oils, bark, and other plant sources. Sterols are a subgroup of steroidal compounds having a hydroxyl group at C-3. Typically, phytosterols have a double bond within the steroid nucleus, such as cholesterol; however, phytosterols may also contain a substituted side chain (R) at C-24, such as an ethyl or methyl group, or another double bond. The structures of phytosterols are well known to those skilled in the art.

[0266] At least 44 naturally occurring phytosterols have been discovered, and they are generally derived from plants such as corn, soybean, wheat, and tung oil; however, they can also be produced synthetically to form compositions identical to those naturally occurring ones or compositions having properties similar to those of naturally occurring phytosterols. According to specific embodiments of the invention, non-limiting examples of phytosterols well known to those skilled in the art include 4-demethylsterols (e.g., β-sitosterol, campesterol, stigmasterol, brassosterol, 22-dehydrobrassosterol, and Δ5-aprilsterol), 4-monomethylsterols, and 4,4-dimethylsterols (triterpenoids) (e.g., cycloartenol, 24-methylenecycloargentanol, and cyclobranol).

[0267] As used herein, the phrases “sterol,” “phytosterol,” and “plant sterol” are synonymous. Phytosterols are saturated sterols that exist in only trace amounts in nature and can also be produced synthetically, such as by hydrogenation of phytosterols. According to specific embodiments of the invention, non-limiting examples of phytosterols include β-sitosterol, campesterol, cycloartenol, and other saturated forms of triterpenoid alcohols.

[0268] Phytosterols and phytosterols as used herein include a variety of isomers such as α and β isomers (e.g., α-sitosterol and β-sitosterol, which respectively comprise one of the most effective phytosterols and phytosterols for lowering serum cholesterol in mammals).

[0269] The phytosterols and phytosterols of the present invention may also be in their ester forms. Suitable methods for obtaining esters of phytosterols and phytosterols 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 Publication No. 2003 / 0045473, the disclosures of which are incorporated herein by reference in their entirety. Non-limiting examples of suitable esters of phytosterols and phytosterols include sitosterol acetate, sitosterol oleate, stigmasterol oleate, and their corresponding phytosterol esters. The phytosterols and phytosterols of the present invention may also include their derivatives.

[0270] Typically, the amount of functional ingredient in a composition varies widely depending on the specific composition and the desired functional ingredient. Those skilled in the art will readily determine the appropriate amount of functional ingredient for each composition.

[0271] In one embodiment, a method for preparing the composition includes combining at least one mogroside of the present invention with at least one sweetener and / or additive and / or functional ingredient.

[0272] VI. Consumer Goods

[0273] The present invention also provides a consumer product comprising at least one mogroside of the present invention or a composition comprising at least one mogroside of the present invention. In one specific embodiment, the at least one mogroside is isolated and purified.

[0274] One or more mogrosides of the present invention, or compositions containing said mogrosides, can be compounded with any known edible or oral compositions (referred to herein as “consumer products”). As used herein, a consumer product means a substance that comes into contact with the mouth of a human or animal, including substances that are ingested and subsequently expelled from the mouth and substances that are consumed, eaten, swallowed, or otherwise ingested, and are healthy for human or animal consumption when used in a generally acceptable manner.

[0275] Exemplary consumer products include pharmaceutical compositions, edible gel mixtures and compositions, dental compositions, foods (sweeteners, condiments, chewing gum, cereal compositions, baked goods, dairy products, and tabletop sweetener compositions), beverages, and beverage products.

[0276] For example, beverages are consumer products. The beverage may be sweetened or unsweetened. One or more mogrosides of the present invention, or compositions containing said mogrosides, may be added to a beverage or beverage base to sweeten the beverage or enhance its existing sweetness or flavor.

[0277] In one specific embodiment, the consumer product contains at least one mogroside of the present invention at a concentration greater than about 1 ppm, such as 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.

[0278] The consumer product may optionally contain additives, additional sweeteners, functional ingredients, and combinations thereof, as described herein. Any of the additives, additional sweeteners, and functional ingredients described above may be present in the consumer product.

[0279] In one embodiment, the composition is a consumer product. In another embodiment, the consumer product is a beverage or beverage product. The beverage or beverage product contains at least one mogroside of the present invention, or a composition containing at least one mogroside of the present invention.

[0280] As used herein, “beverage product” refers to ready-to-drink beverages, beverage concentrates, beverage syrups, or powdered beverages. Suitable ready-to-drink beverages include both carbonated and non-carbonated beverages. Carbonated beverages include, but are not limited to, frozen carbonated beverages, enhanced effervescent beverages, cola, fruit-flavored effervescent beverages (e.g., lemon-lime, orange, grape, strawberry, and pineapple), ginger ale, soft drinks, and sarsaparilla. Non-carbonated beverages include, but are not limited to, fruit juices, fruit-flavored fruit juices, fruit juice drinks, nectar, vegetable juices, vegetable-flavored juices, sports drinks, energy drinks, enhanced water drinks, vitamin-enriched enhanced water, near-water drinks (e.g., water with natural or synthetic flavorings), coconut juice, tea drinks (e.g., black tea, green tea, red tea, oolong tea), coffee, cocoa drinks, beverages containing dairy components (dairy drinks, coffee containing dairy components, café au lait, milk tea, fruit milk drinks), beverages containing grain extracts, drinkable yogurt, protein drinks, and smoothies.

[0281] Beverage concentrates and beverage syrups are prepared using an initial volume of liquid matrix (e.g., water) and desired beverage ingredients. Full-strength beverages are then prepared by adding an additional volume of water. Powdered beverages are prepared by dry-mixing all beverage ingredients in the absence of a liquid matrix. Full-strength beverages are then prepared by adding the full volume of water.

[0282] The beverage contains a matrix, which is the basic component of the dissolved ingredients (including the compositions of the present invention). In one embodiment, the beverage contains beverage-quality water as a matrix, such as deionized water, distilled water, reverse osmosis water, carbon-treated water, purified water, softened water, and combinations thereof. Other suitable matrices include, but are not limited to, phosphoric acid, phosphate buffer, citric acid, citrate buffer, and carbon-treated water.

[0283] Beverages or beverage products may further include at least one additional sweetener and / or functional ingredient and / or additive as described herein.

[0284] Considering that the pH of the composition (e.g., a beverage) does not substantially or adversely affect the taste of the sweetener, a non-limiting example of the pH range of the beverage can be from about 1.8 to about 10. Another example includes a pH range from about 2 to about 5. In one specific embodiment, the pH of the beverage can be from about 2.5 to about 4.2. Those skilled in the art will understand that the pH of the beverage can vary depending on the type of beverage. For example, dairy beverages can have a pH greater than 4.2.

[0285] The titratable acidity of a beverage can range, for example, from about 0.01% to about 1.0% by weight of the beverage.

[0286] In one embodiment, the effervescent beverage product has an acidity of from about 0.01% to about 1.0% by weight of the beverage, such as from about 0.05% to about 0.25% by weight of the beverage.

[0287] The carbonation of the effervescent beverage product has a carbon dioxide content of 0 to about 2% (w / w) or its equivalent, for example from about 0.1% to about 1.0% (w / w).

[0288] The beverage may be caffeinated or decaffeinated.

[0289] The temperature range of the beverage can be, for example, from about 4°C to about 100°C, such as from about 4°C to about 25°C.

[0290] The beverage can be a high-calorie drink, with up to about 120 calories per 8-ounce serving.

[0291] The beverage can be a medium-calorie drink, with up to about 60 calories per 8-ounce serving.

[0292] The beverage can be a low-calorie drink, with up to about 40 calories per 8-ounce serving.

[0293] The beverage can be a zero-calorie drink, which has less than about 5 calories per 8-ounce serving.

[0294] In one specific embodiment, the composition is a cola beverage. The cola beverage may be a low-calorie, medium-calorie, or zero-calorie beverage.

[0295] In one specific embodiment, the beverage is a low-calorie (diet) beverage. In a more specific embodiment, the beverage is a low-calorie carbonated beverage.

[0296] In one specific embodiment, the beverage of the present invention is a flavored water beverage.

[0297] The concentration of mogroside of the present invention in beverages can be higher than, equal to or lower than the threshold sweetness or flavor recognition concentration of mogroside of the present invention.

[0298] In one embodiment, the mogroside of the present invention is present in the beverage at a concentration higher than about 1 ppm, for example, about 1 ppm to about 1,000 ppm, about 25 ppm to about 1,000 ppm, about 50 ppm to about 1,000 ppm, about 75 ppm to about 1,000 ppm, about 100 ppm to about 1,000 ppm, about 200 ppm to about 1,000 ppm, about 300 ppm to about 1,000 ppm, about 400 ppm to about 1,000 ppm, or about 500 ppm to about 1,000 ppm.

[0299] In a more specific embodiment, the mogroside of the present invention is present in amounts from about 25 ppm to about 600 ppm, for example, from about 25 ppm to about 500 ppm, from about 25 ppm to about 400 ppm, from about 25 ppm to about 300 ppm, from about 25 ppm to about 200 ppm, from about 25 ppm to about 100 ppm, from about 50 ppm to about 600 ppm, from about 50 ppm to about 500 ppm, from about 50 ppm to about 400 ppm, from about 50 ppm to about 300 ppm, from about 50 ppm to about 200 ppm, from about 50 ppm to about 100 ppm, from about 100 ppm to about 600 ppm, from about 100 ppm. It exists in beverages at concentrations of about 500 ppm, about 100 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.

[0300] In one embodiment, the beverage has a sweetness of 3 Brix or greater, such as 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.

[0301] In another embodiment, the mogroside of the present invention is present in the beverage in an amount that effectively provides 3 Brix or greater, such as 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.

[0302] In yet another embodiment, the sweetener composition of the present invention is present in a beverage in an amount that effectively provides 3 or more Brix, such as 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, or 10 or more Brix.

[0303] VII. Instructions for Use

[0304] The compounds and compositions of the present invention can be used to impart sweetness to consumer products or enhance their flavor or sweetness. In specific embodiments, one or more mogrosides of the present invention can be used to sweeten consumer products and / or enhance their flavor. Compared to consumer products without at least one mogroside of the present invention, consumer products containing at least one mogroside of the present invention have a more sucrose-like taste profile.

[0305] In one aspect, the present invention is a method for preparing a sweetened consumer product, the method comprising (i) providing the consumer product and (ii) adding at least one mogroside of the present invention to the consumer product to provide the sweetened consumer product.

[0306] In one specific embodiment, a method of preparing a sweetened consumer product includes (i) providing an unsweetened consumer product and (ii) adding at least one mogroside of the present invention to the unsweetened consumer product to provide a sweetened consumer product.

[0307] In one specific embodiment, 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 to provide a sweetened beverage.

[0308] In one specific embodiment, the present invention is a method for preparing a sweetened beverage, the method 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.

[0309] In the above methods, one or more mogrosides of the present invention can be provided as is, i.e., in the form of compounds, or in the form of compositions. When provided as compositions, the amount of mogrosides in the composition effectively provides a mogroside concentration higher than, equal to, or lower than its flavor or sweetness recognition threshold when the composition is added to a consumer product (e.g., a beverage). When one or more mogrosides of the present invention are not provided as compositions, they can be added to consumer products at a concentration higher than, equal to, or lower than their flavor or sweetness recognition threshold.

[0310] In one embodiment, the present invention is a method for enhancing the sweetness of a consumer product, the method comprising (i) providing a consumer product containing at least one sweetener in an amount of sweetening agent and (ii) adding at least one mogroside of the present invention or a composition containing said mogroside to said consumer product to provide a consumer product with enhanced sweetness, wherein the mogroside of the present invention is added to said consumer product at a concentration equal to or below its sweetness recognition threshold. In one specific embodiment, the mogroside of the present invention is added to the consumer product at a concentration below its sweetness recognition threshold.

[0311] In one specific embodiment, the present invention is a method for enhancing the sweetness of a beverage, the method comprising (i) providing a beverage containing at least one sweetener in an amount of sweetening agent and (ii) adding at least one mogroside of the present invention or a composition containing said mogroside to said beverage to provide a beverage with enhanced sweetness, wherein said mogroside is added to said beverage at a concentration equal to or below its sweetness recognition threshold. In one specific embodiment, the mogroside of the present invention is added to a consumer product at a concentration below its sweetness recognition concentration threshold.

[0312] In another embodiment, the present invention is a method for enhancing the flavor of a consumer product, the method comprising (i) providing a consumer product containing at least one flavor component and (ii) adding at least one mogroside of the present invention or a composition containing said mogroside to said consumer product to provide a consumer product with enhanced flavor, wherein the mogroside of the present invention is added to said consumer product at a concentration equal to or below its flavor recognition threshold. In one specific embodiment, the mogroside of the present invention is added to the consumer product at a concentration below its flavor recognition threshold.

[0313] In one specific embodiment, a method for enhancing the flavor of a beverage is provided, the method comprising (i) providing a beverage containing at least one flavor component and (ii) adding at least one mogroside of the present invention or a composition containing said mogroside to said beverage to provide a beverage with enhanced flavor, wherein said mogroside is added to said beverage at a concentration equal to or below a flavor recognition threshold of said mogroside. In one specific embodiment, the mogroside of the present invention is added to a consumer product at a concentration below its flavor recognition threshold.

[0314] In one embodiment, a method of enhancing the sweetness of a consumer product includes (i) providing a consumer product containing at least one sweetener and (ii) adding at least one mogroside of the present invention to a composition to provide a composition having enhanced sweetness.

[0315] In another embodiment, a method for enhancing the sweetness of a consumer product includes (i) providing a consumer product matrix and (ii) adding at least one sweetener and at least one mogroside of the present invention to the consumer product matrix to provide a consumer product with enhanced sweetness. The at least one sweetener and the at least one mogroside of the present invention may be added together, i.e., added as a composition, or added separately.

[0316] As used herein, the term “consumer product matrix” refers to a composition containing all of the typical ingredients except for sweeteners or sweetener compositions.

[0317] In one specific embodiment, the SE of a consumer product comprising at least one mogroside of the present invention and at least one sweetener is enhanced by at least about 1.2 times compared to the SE of a consumer product in the absence of at least one mogroside of the present invention, such as at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, at least about 1.6 times, at least about 1.7 times, at least about 1.8 times, at least about 1.9 times, and at least about 2.0 times.

[0318] In another embodiment, compared with the Brix content of a consumer product that does not contain mogrosides of the present invention, adding at least one mogroside of the present invention to a consumer product or a consumer product matrix increases the Brix content by at least 1 degree, such as at least 2 degrees, at least 3 degrees, or at least 4 degrees.

[0319] In another aspect, a method for making a consumer product taste more like sucrose-sweetened products includes (i) providing a consumer product containing at least one sweetener in an amount that enhances sweetness and (ii) adding at least one mogroside of the present invention to effectively adjust one or more flavor properties of the sweetener to make the consumer product taste more like sucrose-sweetened products compared to a consumer product in the absence of at least one mogroside of the present invention.

[0320] In another embodiment, a method for making a consumer product taste more like sucrose-sweetened food includes (i) providing a consumer product matrix and (ii) adding at least one sweetener and at least one mogroside of the present invention to the consumer product matrix to provide a consumer product that tastes more like sucrose-sweetened food, wherein the at least one mogroside of the present invention is present to effectively modulate one or more flavor properties of the sweetener to make the consumer product taste more like sucrose-sweetened food compared to a consumer product 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., added in the form of a composition, or added separately.

[0321] A method for preparing compositions with enhanced sweetness is also provided.

[0322] In one aspect, a method of preparing the 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.

[0323] In one aspect, the present invention is a method for preparing a sweetened consumer product, the method comprising (i) providing the consumer product and (ii) adding at least one mogroside of the present invention to the consumer product in a sweetening amount to provide the sweetened consumer product.

[0324] In one specific embodiment, a method of preparing a sweetened consumer product includes (i) providing an unsweetened consumer product and (ii) adding at least one mogroside of the present invention to the unsweetened consumer product in a sweetening amount to provide a sweetened consumer product.

[0325] In one specific embodiment, the present invention is a method for preparing a sweetened beverage, the method comprising (i) providing the beverage and (ii) adding at least one mogroside of the present invention to the beverage in a sweetening amount to provide the sweetened beverage.

[0326] In one specific embodiment, the present invention is a method for preparing a sweetened beverage, the method 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 a sweetened beverage.

[0327] VIII. Purification Methods

[0328] The present invention is also extended to the method for purifying mogroside of the present invention.

[0329] In one embodiment, the present invention is a method for purifying mogrosides of the present invention, the method comprising (i) passing a solution containing a source material containing mogrosides of the present invention through an HPLC column, and (ii) eluting a fraction containing mogrosides of the present invention to provide a purified mogroside composition containing mogrosides of the present invention. The HPLC column can be any suitable preparative or semi-preparative HPLC column.

[0330] As used herein, the term "preparative HPLC" refers to an HPLC system capable of producing product fractions in the sizes of high (500 or more) micrograms, milligrams, or grams. The term "preparative" includes both preparative columns and semi-preparative columns, but is not intended to include analytical columns that provide fractions in the nanogram to low microgram range.

[0331] As used herein, an "HPLC-compatible detector" is a detector suitable for use in an HPLC system that provides a detectable signal after the elution of a compound peak. For example, a detector that generates a signal when a compound is eluted is an HPLC-compatible detector. In cases where the absorbance of a component varies widely, it may be necessary to use more than one detector. Since an "incompatible" detector cannot detect undesirable peaks, a detector that can detect the desired component is not an "incompatible" detector.

[0332] An HPLC apparatus typically includes at least the following components: 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 eluted from the column. The apparatus may optionally include components for providing gradient elution, although this is not required when using the methods described herein. Conventional methods and equipment for performing HPLC separations are well known in the art.

[0333] A suitable stationary phase is one in which the compound of interest is eluted. Preferred columns can be, but are not limited to, normal-phase columns (neutral, acidic, or basic), reverse-phase columns (with alkyl chains of any 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 ranges from a few μm to hundreds of μm.

[0334] Suitable detection devices include, but are not limited to, mass spectrometers, UV detectors, IR detectors, and light scattering detectors. The methods described herein utilize any combination of these detectors. The most preferred embodiment uses a mass spectrometer and a UV detector.

[0335] As used herein, "source material" refers to material purified by the method of the present invention. The source material contains mogrosides of the present invention, but at a purity lower than that provided by the purification method of the present invention. The source material may be liquid or solid. Exemplary source materials include, but are not limited to, mixtures of mogrosides and mogroside extracts (commercial or prepared).

[0336] As those skilled in the art will understand, any solid source material must be introduced into the solution before performing an HPLC method.

[0337] In one embodiment, a representative analytical HPLC protocol is associated with a preparative or semi-preparative HPLC protocol used to purify compounds.

[0338] In another embodiment, appropriate conditions for purifying the mogrosides of the present invention can be determined by identifying representative samples for a given analytical HPLC column, solvent system, and flow path. In yet another embodiment, relevant preparative or semi-preparative HPLC methods can be applied to purify the mogrosides of the present invention, wherein the purification parameters may or may not be changed.

[0339] 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.

[0340] 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.

[0341] In one embodiment, impurities are eluted from the HPLC column after eluting one or more fractions containing mogrosides of the present invention. In another embodiment, impurities are eluted from the HPLC column before eluting one or more fractions containing mogrosides of the present invention.

[0342] The method may further include removing the solvent from the elution solution, i.e., drying. In one embodiment, the method further includes partially removing the solvent from the elution solution to provide a concentrate containing mogrosides of the present invention. In another embodiment, the method further includes removing substantially all of the solvent from the elution solution to provide a substantially dry composition containing mogrosides of the present invention.

[0343] Solvent removal can be carried out by any means known to those skilled in the art, including but not limited to evaporation, distillation, lyophilization, vacuum drying and spray drying.

[0344] The purified fraction containing mogrosides of the present invention can be further purified by other methods to improve 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.

[0345] The source material can be one fraction or multiple fractions containing the mogrosides of the present invention collected by at least one prior method or HPLC protocol. In one embodiment, multiple fractions from the same prior method or HPLC protocol are combined, and optionally, the solvent is removed before subjecting the source material to another method. In other embodiments, fractions from different prior methods or HPLC protocols are combined, and optionally, the solvent is removed before subjecting the source material to another method.

[0346] In one embodiment, the source material subjected to one or more additional methods comprises a liquid fraction obtained from one or more prior (and optionally, different) methods, which is mixed with a substantially dry material obtained by drying a fraction obtained from one or more prior (and optionally, different) methods. In another embodiment, the source material subjected to one or more additional methods comprises a substantially dry material obtained by drying a fraction obtained from one or more prior (and optionally, different) methods, wherein the source material is introduced into a solution, which is then passed through a next HPLC column.

[0347] The second and subsequent methods can have different HPLC schemes (e.g., solvent system, column, method) and different post-elution steps (e.g., partial solvent removal, complete solvent removal, elution of impurities, crystallization, or extraction).

[0348] The separated material can be subjected to two, three, four or more further processes, each time providing a higher level of purified mogroside of the present invention.

[0349] In one embodiment, the method provides a purified mogroside composition comprising the mogroside of the present invention, having a purity of at least about 80% or higher by weight, such as at least about 85%, at least about 90%, at least about 95%, or at least about 97% or higher by weight. In another embodiment, purification provides pure mogroside of the present invention, i.e., >99% by weight on a dry basis.

[0350] The advantages of the present invention will become more apparent from the detailed description given below. However, it should be understood that while pointing out preferred embodiments of the invention, the detailed description and specific examples are given only by way of illustration, as 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.

[0351] Example

[0352] Example 1: Biotransformation of mogroside V

[0353] LC-MS was performed on MV 90 (90% mogroside V) substrate (Hunan Huacheng Biotech Inc.). Two significant peaks were observed in the LC-MS of the mass extracts of xylose-containing mogrosides with 5-glycosylated + / - sodium (“xylose-containing mogroside 1” and “xylose-containing mogroside 2”). The results are shown in Figure 1.

[0354] When using the DSM Maxilact enzyme in a biotransformation reaction, the following reaction is expected to occur.

[0355] Option 1

[0356]

[0357] In one embodiment, a reaction is provided to convert a xylose-containing compound, such as isomogroside V, into a xylose-containing compound, such as mogroside IV.

[0358] Option 2

[0359]

[0360] One or two of these can be converted into mogrosides containing xylose with four glycosylations.

[0361] Based on our understanding of the structure of mogrosides containing 5-glucosylation (i.e., mogrosides with only glucose), it is highly likely that xylose-containing mogrosides with 5 glycosyl units are similar to mogroside V and isomogroside V.

[0362] Example 2: Diversification of mogrosides

[0363] To prepare mogroside compounds using the biotransformation pathway of mogroside V, various enzymes for adding non-glucose monosaccharides to the C24 glycosyl group of MogIIIE were screened.

[0364] Option 3:

[0365]

[0366] Materials and instruments:

[0367] Select glycosyltransferases and transglycosylation glycosidases capable of adding α / β non-glucose sugars to glucose positions (not only C6, but also C3 and C4). Enzymes that can be used for inversion reactions 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).

[0368] Reaction mixture:

[0369] 50 μL reaction (40℃ for 40 hours):

[0370]

[0371] Setting up a pH step to assess changes in biotransformation yield:

[0372] Acetate buffer 3.6;

[0373] Acetate buffer 4.6;

[0374] Acetate buffer 5.6;

[0375] Phosphate buffer 6.6;

[0376] Phosphate buffer 7.0

[0377] We used the two tables above to prepare buffer solutions with different pH values ​​(4.6, 5.6, 6.6).

[0378] Purification process

[0379] The reaction mixture was purified by passing it through a C18 column using an HPLC system.

[0380] 1. Rhamnose

[0381] 25 mL reaction (40°C for 60 hours):

[0382] α-Rhamnosidase 0.5 mL

[0383] 12.5 mL of 500 mM rhamnose in 2× buffer, pH 5.6

[0384] 130mM MogIIIE 11mL

[0385] Add 0.5 mL of α-rhamnosidase at 30 h.

[0386] After 60 hours, the reaction was boiled, filtered, and purified by HPLC. This... Figure 2 As shown in the image.

[0387] After C18 HPLC purification (10%-40% acetonitrile), a peak of approximately 24 mg was collected at peak 45. Peak 46 consisted of two indistinguishable compounds. Peaks 47 and 48 were the substrate MogIIIE. Peak 45 was subjected to NMR analysis and identified as CC-00500.

[0388] The reaction at pH 4.6–5.6 has a high productivity.

[0389] The separated HPLC peaks were subjected to mass spectrometry analysis, as shown in Figure 3. Both separated HPLC peaks showed an increase in mass from 985 to 1131, as expected with the addition of one rhamnose molecule to the mogroside IIIE substrate.

[0390] 2. Galactose

[0391] 25 mL reaction (40°C for 60 hours):

[0392] α-Galactosinase 0.5 mL

[0393] 12.5 mL of 500 mM rhamnose in 2× buffer, pH 4.6

[0394] 130mM MogIIIE 11mL

[0395] Add 0.5 mL of α-galactosidase at 30 h.

[0396] After 60 hours, the reaction was boiled and filtered, then purified by HPLC. This... Figure 4 As shown in the image.

[0397] After C18 HPLC purification (10%-40% acetonitrile), a peak of approximately 4 mg was collected at 108, a peak of approximately 28 mg was collected at 109, and peak 110 contained a large amount of the substrate MogIIIE. Peaks 111 and 112 were also composed of the substrate MogIIIE. Peaks 108 and 109 were subjected to NMR analysis, and peak 108 was identified as CC-00489.

[0398] The reaction at pH 3.6–4.6 exhibited high productivity. The separated HPLC peaks were subjected to mass spectrometry analysis, as shown in Figure 5. Both separated HPLC peaks showed a mass increase from 985 to 1147, as expected with the addition of one rhamnose molecule to the mogroside IIIE substrate.

[0399] 25 mL reaction (40°C for 60 hours):

[0400] α-Galactosinase 0.5 mL

[0401] 500 mM xylose in 2× buffer, pH 4.6, 12.5 mL

[0402] 130mM MogIIIE 11mL

[0403] Add 0.5 mL of α-galactosidase at 30 h.

[0404] After 60 hours, the reaction was boiled and filtered, then purified by HPLC. The reaction at pH 4.6 exhibited high productivity. This... Figure 6 As shown in the image.

[0405] After C18 HPLC purification (10%-40% acetonitrile), peaks 36, 37, 38, 39, and 40 were collected. Mass spectrometry analysis of peak 39 showed a secondary peak with m / z of 1117, indicating that xylose had been added to mogroside IIIE, such as... Figure 7 As shown in the image.

[0406] in conclusion:

[0407] MALDI-TOF analysis showed that adding galactose, rhamnose, or xylose resulted in higher productivity.

[0408] Example 3: Sweetness rating of CC-00489

[0409] The sweetness of mogroside-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 reference.

[0410] Detailed description:

[0411] 1. Reference material: 8%, 9%, and 10% sucrose in deionized water.

[0412] 2. Measure the sweetness of the sample relative to a sucrose reference.

[0413] 3. Testing method: sipping and spitting out.

[0414] 4. Other temporal characteristics: bitterness, astringency, lingering sweetness, and bitter aftertaste.

[0415] 5. The test level for CC-00489 is 400 ppm.

[0416] 6. The batch number of CC-00489 is IN-SDV-D-196-2.

[0417] Sample preparation

[0418] Table 1: CC-00489 at 400 ppm in deionized water

[0419] Element quantity CC-00489 2.5mg Deionized water 6.25g

[0420] Table 2: Sucrose in deionized water

[0421] Element 8% sucrose 9% sucrose 10% sucrose sucrose 8g 9g 10g Deionized water 92g 91g 90g

[0422] Add the ingredients to deionized water while stirring until the solid dissolves visibly. Pour the sample into a glass bottle or vial and store at 4°C.

[0423] Taste Review

[0424] Taste testing was conducted by two expert panel members. Bottles / vials were removed from the refrigerator. Approximately 25 ml of the sucrose solution was poured into a 4-ounce plastic cup, and 3 ml of the sweetener solution into a 2-ounce plastic cup. Before tasting and between tasting different samples, the expert panel members were given mineral water to rinse their mouths. Before tasting the next sample, unsalted biscuits were also given to the expert panel members, followed by rinsing their mouths with mineral water.

[0425] Initially, the expert panel was asked to examine the sweetness of three sucrose solutions in deionized water (Table 2).

[0426] Samples of compound A in deionized water (Table 1) were given to the panel members. Panel members were asked to measure the sweetness of the sample compared to a sucrose reference (Table 2) and to describe any taste characteristics, such as bitterness, astringency, and bitter aftertaste. Panel members were instructed to sip the sample, evaluate the sweetness, and then spit the sample into a cup provided for this purpose.

[0427] One panel member assessed the sweetness of the sample containing compound A as 9-10 sucrose equivalents (SE), noting a slightly astringent taste and a pleasant mouthfeel. Another panel member assessed the sweetness of the sample as close to 10 SE.

[0428] Example 4: Purification using C18 resin

[0429] After the substrate of MV 90 (90% mogroside V) (Hunan Huacheng Biotech Inc.) was bioconverted to sympathoside I using a β-galactosidase (e.g., Aspergillus oryzae β-galactosidase (AoBG), such as Maxilact A4), the mogroside of interest (e.g., CC-00520) was isolated.

[0430] Dissolve 340g of MV 90 in 1L of potassium phosphate buffer (100mM, pH 6.5) and dilute with 1L of water (total 2L, final buffer concentration 50mM). Then filter through a 0.2μm sterile filter unit.

[0431] 6 L of freshly concentrated Maxilact A4 (3.5× concentrate) was diluted with 6 L 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 barrel equipped with coarse glass frit. The filtrate was passed through a series of capsule filters (5 μm and 0.2 μm) and finally filtered into a sterile filter unit. The final volume of the filtered enzyme was approximately 11 L.

[0432] 11 L of sterile filter enzyme and 2 L of sterile Mog. V solution were independently transferred to the fermenter. The pH was adjusted to 6.2 and the reaction temperature was set to 50 °C, and the reaction was initiated.

[0433] Following the biotransformation reaction, the mogrosides of the present invention are separated from the enzymes and salts. To separate the protein from the mogrosides, the reaction mixture is mixed with sodium hydroxide to increase the pH to 12.4. Ethanol is added to prepare a 20% ethanol solution. The mixture is filtered through a 10 kDa Koch Romicon membrane with an inlet pressure of 1.7 barg and an outlet pressure of atmospheric pressure. The pH of the permeate is lowered to 5.5 using acetic acid and cooled overnight. The next day, the solution is re-filtered through a 10 kDa Koch Romicon membrane.

[0434] Water, ethanol, and salt were removed using a Koch SR3D nanofiltration membrane with a rejection capacity of 200 Da. The solution was percolated until the ethanol concentration was below 3%, and then concentrated to 20-30 L. The concentrated mogrosides were then mixed with a water / ammonia acetate solution to bring the total solution volume to approximately 110 L.

[0435] Pass the mixture through a Biotage SNAP KP-C18-HS 400g guard column. Mix the resulting solutions and dispense them into equal portions into 5L HDPE jerry cans.

[0436] Chromatography

[0437] Chromatographic separation has six stages:

[0438] 1) Column balancing to prepare for load.

[0439] 2) Column loading. After loading, a small amount of equilibration solution is applied to distribute mogrosides throughout the bed.

[0440] 3) Remove mogroside V and other compounds eluted earlier. Take the first approximately 120 L sample and discard it. Collect the next approximately 27 kg; the target purity appears in the final fraction.

[0441] 4) Removal of sialoside I (two large 18kg fractions), followed by 4 × 4.5kg fractions. The final fraction did not meet purity specifications.

[0442] 5) Pure XNS grade ethanol (95%). The first 18 kg collected was of Mog IIIe grade.

[0443] 6) Use pure XNS grade ethanol to clean the column.

[0444] The composition of the eluent is shown in Table 14.

[0445] Table 14: Column Eluent Mixing

[0446]

[0447] percolation

[0448] Further processing is performed on one or more fractions containing mogrosides of the present invention. Fractions containing mogrosides of the present invention are identified by, for example, HPLC-MS, and typically eluted with symmonidine I. Ethanol and ammonium acetate are removed. The combined fractions are percolated and RO water is added to the solution to maintain an ethanol concentration below 15%. Ammonia, acetate, ethanol, and water are removed.

[0449] Secondary HPLC chromatography was performed, for example, on another C18 column, to further purify the mogrosides of the present invention. The fractions were freeze-dried for analysis and storage.

[0450] Example 5: Synthesis of CC-00491

[0451] 3-O-β-D-glucopyranosylmogroside 24-O-β-D-glucopyranosyl-(1→2)-α-L-rhamnosyl-(1→6)-β-D-glucopyranosyl (Scheme 1). At room temperature under N2 atmosphere, a 0.5 M solution (0.24 mL, 0.12 mmol) of NaOMe in MeOH was added dropwise to a solution of 6 (562.5 mg, 0.24 mmol) in dry THF (10 mL) and dry MeOH (10 mL). The mixture was stirred at the same temperature for 18 h, neutralized with 10% AcOH aqueous solution, and concentrated under reduced pressure. The remaining white solid was subjected to preparative HPLC (Phenomenex Luna C18 column, 250 × 30 mm, 10 μm particles, H2O → CH3CN gradient, 40 mL / min, t). R Purification was performed at a retention time of 11-15 min. Fractions 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. mp = 179℃-182℃R f =0.56 (silica gel, CH2Cl2 / MeOH / H2O = 5:4:1). 11H NMR (500 MHz, pyridine-d5) δ 7.53 (d, J = 5.0 Hz, 1H), 7.51 (d, J = 3.0 Hz, 1H), 7.44 (d, J = 3.0 Hz, 1H), 7.30 (d, J = 3.8 Hz, 1H), 7.19 - 7.12 (m, 2H), 7.11 - 6.99 (m, 2H), 6.65 - 6.53 (m, 2H), 6.46 (d, J = 6.2 Hz, 1H), 6.22 - 6.13 (m, 1H), 5.66 (d, J = 5.8 Hz, 1H), 5.47 (d, J = 6.4 Hz, 1H), 5.46 (d, J = 1.1 Hz, 1H), 5.43 (t, J = 5.9 Hz, 1H), 5.36 (d, J = 7.8 Hz, 1H), 5.16 (s, 1H), 5.04 (d, J = 7.8 Hz, 1H), 4.91 (d, J = 7.8 Hz, 1H), 4.70 - 4.65 (m, 1H), 4.64 (d, J = 9.7 Hz, 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 = 6.2 Hz, 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 = 6.4 Hz, 3H), 0.90 (s, 3H), 0.89 (s, 3H). 13C NMR (125MHz, 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.

[0452]

[0453] Example 6: Purification and Characterization of CC-00507

[0454] Materials: The material used to separate CC-00507 (lot number #IN-RAS-A-53-3) was Luo Han Guo extract (lot number #LHGE-180125) purchased from Huacheng Bio.

[0455] HPLC analysis: HPLC analysis was performed on an Agilent 1200 system coupled with a variable wavelength detector (VWD). Sample processing and final purity evaluation were performed using the conditions described in Table 1.

[0456] Table 1: Analytical HPLC conditions used for final purity evaluation

[0457]

[0458] Preparative HPLC analysis: Preparative HPLC analysis was performed on an Agilent preparative HPLC system coupled with a UV-Vis detector.

[0459] Primary processing: Approximately 109 g of *Monk Fruit* extract (lot number #LHGE-180125) was treated using the preparative HPLC method described in Table 2. The retention time of the target fraction was 20.00–23.00 min (peak ID: LHGE-180125-P4). This material was combined and lyophilized. The final yield of peak ID: LHGE-180125-P4 (lot number #IN-RAS-A-24-4) was approximately 12.2 g.

[0460] Table 2: Preparative HPLC method conditions for primary treatment of #LHGE-180125.

[0461]

[0462]

[0463] 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 minutes (peak ID: LHGE-180125-P4-D). This material was combined and lyophilized. The final yield of peak ID: LHGE-180125-P4-D (lot number #IN-VVP-K-194-4) was approximately 60 mg.

[0464] Table 3: Preparative HPLC Method Conditions for Processing #LHGE-180125-P4

[0465]

[0466]

[0467] Third-stage 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). This material was combined and lyophilized for separation. The final yield of peak ID: LHGE-180125-P4-D3 (lot number #IN-RAS-A-53-3) was 4.4 mg with a purity of 90.6% (area %).

[0468] Table 4: Preparative HPLC Method Conditions for Processing #LHGE-180125-P4-D

[0469]

[0470]

[0471] MS and MS / MS. MS and MS / MS data were generated using a Waters QTof Micro mass spectrometer equipped with an electrospray ionization source. Samples were analyzed by negative ESI. The sample (approximately 0.2 mg) was diluted to approximately 0.2 mg / mL with 50:50 ACN:H2O and introduced via direct perfusion.

[0472] Mass spectrometry. ESI-TOF mass spectra obtained by perfusing the CC-00507 sample showed [MH] at m / z 1255.6326.- Ions. [MH] - The mass of the ion and the expected molecular formula C 59 H 100 O 28 Very consistent (for C) 59 H 99 O 28 The calculated value is 1255.6323 (error: 0.2 ppm). MS data confirms that CC-00507 has a nominal mass of 1256 Daltons and a molecular formula of C2. 59 H 100 O 28 The ion observed at m / z 1353.6130 is most likely attributed to [M-H+H3PO4]. - .

[0473] MS / MS spectrum of CC-00507 (selected [MH] at m / z 1255.5) - Ion fragmentation indicates the loss of a xylose unit at m / z 1123.5382, followed by the successive loss of four glucose units at m / z 961.4853, 799.4445, 637.3991, and 475.3554. Following the loss of xylose units from the structure, an alternative fragmentation pathway was also observed in the spectrum, corresponding to the loss of a water molecule from the central triterpene core, followed by the successive loss of sugar units at m / z 1105.5109, 943.4888, 781.4421, and 619.3909.

[0474] NMR. Samples were prepared by dissolving available materials in 130 μL of CD3OD, and NMR data were acquired. Data were obtained on a Bruker Avance 500 MHz NMR instrument equipped with a 2.5 mm reverse probe. 1 H, 1 H- 1 H COSY、 1 H- 13 C HSQC-DEPT and 1D TOCSY NMR data. Due to the low sample concentration, additional NMR data, such as [data missing], were acquired at Rensselaer Polytechnic Institute using a Bruker Avance 600MHz instrument equipped with a 5mm cryopreservation probe. 13 C 1 H- 13 C HMBC and 1 H- 1 H ROESY. 1 H NMR spectral reference at δ HThe CHD2OD resonance at 3.30 and 13 C NMR spectral reference at δ C CD3OD resonance at 49.0.

[0475] Table 5. 1 H and 13 C NMR (500 and 125 MHz, CD3OD), designation of CC-00507 aglycone.

[0476]

[0477]

[0478] The terms can be interchanged. ￥ There are two carbon resonances at 26.2 ppm (26.20 ppm and 26.24 ppm), so the chemical shift cannot be definitively specified. There is a partially overlapping methyl resonance at 1.10 ppm (2D NMR data confirm the observation of H-19 at 1.104 ppm and H-26 at 1.097 ppm).

[0479] Table 6. 1 H and 13 C10 NMR (500 and 125 MHz, CD3OD), designation of CC-00507C-3 glycoside.

[0480] Location <![CDATA[ 13 C]]> <![CDATA[ 1 H]]> <![CDATA[Glc IV -1]]> 106.4 4.28d(7.7) <![CDATA[Glc IV -2]]> 75.6 3.19m <![CDATA[Glc IV -3]]> <![CDATA[77.7-78.1 § ]]> Approximately 3.30m <![CDATA[Glc IV -4]]> <![CDATA[71.5-71.6 λ ]]> Approximately 3.30m <![CDATA[Glc IV -5]]> 77.3 3.40m <![CDATA[Glc IV -6]]> 69.7 3.80m, 4.05m <![CDATA[Glc V -1]]> 104.8 4.42d(8.0) <![CDATA[Glc V -2]]> <![CDATA[75.2 € ]]> 3.18m <![CDATA[Glc V -3]]> <![CDATA[77.7-78.1 § ]]> 3.35m <![CDATA[Glc V -4]]> <![CDATA[71.5-71.6 λ ]]> 3.28m <![CDATA[Glc V -5]]> <![CDATA[77.7-78.1 § ]]> Approximately 3.26m <![CDATA[Glc V -6]]> <![CDATA[62.7 ￥ ]]> 3.65m, 3.85m

[0481] § There are six carbon resonances in the range of 77.7–78.1 ppm (77.68 ppm, 77.93 ppm, 78.00 ppm, and 78.14 ppm; two additional carbons overlap in this region), so the chemical shift cannot be definitively specified.

[0482] λ There are four carbon resonances in the range of 71.5–71.6 ppm (71.48 ppm, 71.58 ppm, 71.61 ppm and 71.64 ppm), so the chemical shift cannot be clearly specified.

[0483] € There are two overlapping carbon resonances at 75.2 ppm, so the chemical shift cannot be clearly specified.

[0484] ￥ There are two overlapping carbon resonances at 62.7 ppm, therefore the chemical shift cannot be definitively specified. (Table 7) 1 H and 13CNMR (500 and 125 MHz, CD3OD), designated as CC-00507C-24 glycoside.

[0485] Location <![CDATA[ 13 C]]> <![CDATA[ 1 H]]> <![CDATA[Glc I -1]]> 104.1 4.40d 8.2) <![CDATA[Glc I -2]]> 82.7 3.48m <![CDATA[Glc I -3]]> 78.5 3.57m <![CDATA[Glc I -4]]> <![CDATA[71.5-71.6 λ ]]> 3.33m <![CDATA[Glc I -5]]> 76.4 3.49m <![CDATA[Glc I -6]]> 70.1 3.61m, 4.23m Xyl-1 106.0 4.62d(7.5) Xyl-2 75.8 3.25m Xyl-3 <![CDATA[77.7-78.1 § ]]> 3.32m Xyl-4 71.2 3.49m Xyl-5 67.2 3.15m, 3.84m <![CDATA[Glc II -1]]> 104.4 4.27d(7.7) <![CDATA[Glc II -2]]> <![CDATA[75.2 € ]]> 3.20m <![CDATA[Glc II -3]]> <![CDATA[77.7-78.1 § ]]> 3.35m <![CDATA[Glc II -4]]> <![CDATA[71.5-71.6 λ ]]> Approximately 3.27m <![CDATA[Glc II -5]]> <![CDATA[77.7-78.1 § ]]> Approximately 3.27m <![CDATA[Glc II -6]]> <![CDATA[62.7 ￥ ]]> 3.64m, 3.84m

[0486] λ There are four carbon resonances in the range of 71.5–71.6 ppm (71.48 ppm, 71.58 ppm, 71.61 ppm and 71.64 ppm), so the chemical shift cannot be clearly specified.

[0487] § There are six carbon resonances in the range of 77.7–78.1 ppm (77.68 ppm, 77.93 ppm, 78.00 ppm, and 78.14 ppm; two additional carbons overlap in this region), so the chemical shift cannot be definitively specified.

[0488] € There are two overlapping carbon resonances at 75.2 ppm, so the chemical shift cannot be clearly specified.

[0489] ￥ There are two overlapping carbon resonances at 62.7 ppm, so the chemical shift cannot be clearly specified.

[0490] Spectroscopic (NMR) and spectroscopic (MS) analyses of CC-00507 allowed for the complete specification of its structure, identifying it as mogroside-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 contains four glucose units and one xylose unit. The xylose unit is linked to Glc via a 1→2 glycosidic bond. I .

[0491] Example 7: Preparation and Characterization of CC-00518

[0492] From mogroside III E CC-00518 was isolated from the reaction product of biotransformation with α-D-xylosylfluorine and α-xylosidase (Megazyme, E-AXSEC). The biotransformation was performed under the following conditions: 10 mM α-xylosylfluorine, 10 mM MogIIIE, 0.01% BSA, 6-unit α-xylosidase, in 50 mM phosphate buffer (pH 7.0) at 37°C for approximately 30 min. CC-00518 was purified using preparative HPLC in multiple rounds to provide approximately 25 mg of 99% purity (area %). A series of 1D and 2D experiments were conducted. 1 H, 13 C1 H- 1 H COSY、 1 H- 13 C HSQC-DEPT, 1 H- 13 C HSQC-TOCSY, and 1 H- 13 C HMBC was used to elucidate the structure as mogroside-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]. Spectroscopic data revealed a central triterpenoid core and four sugar units: three glucose units and one xylose unit. The bonds between the sugar units and their attachment to the central triterpenoid core were determined based on COSY, TOCSY, and HSQC-DEPT data. The glucose unit exists as a β-terminal isomer, while the xylose unit (… 3 J = 3.7 Hz exists in the α configuration.

[0493] Table 1. 1 H NMR (500.13MHz, CD3OD) and 13 C NMR (125MHz) is specified as CC-00518.

[0494]

[0495]

[0496]

[0497] Example 8: Preparation, purification and characterization of CC-00520

[0498] CC-00520, a minor mogroside, was isolated from the conversion of mogroside V to symbioside I, as in Example 4. The compound can also be found in and isolated from mogroside extracts.

[0499] Materials: The materials used for separating batch number #IN-VVP-K-172(CC-00520) were R11 purified samples and batch number #AMR100489-23-F2.

[0500] The material used for separating batch number #IN-RAS-A-119-3(CC-00520) was a monk fruit extract sample, batch number #LHGE-180125.

[0501] HPLC analysis: HPLC analysis was performed on an Agilent 1200 system coupled with a variable wavelength (VWD) detector. Multiple procedures and final purity assessments of the samples were performed using the conditions described in Tables 1 and 2.

[0502] Table 1. Analytical HPLC conditions for fractionation analysis in multiple processes and final purity evaluation

[0503]

[0504]

[0505] Table 2. Analytical HPLC conditions for fractionation analysis in multiple processes and final purity evaluation

[0506]

[0507]

[0508] Primary processing: Approximately 68 g of lot number #AMR100489-23-F2 was processed using the primary preparative HPLC method described in Table 3. Fractions with peak ID: AMR100489-23-F2-P2 (lot number #IN-SDV-D-168-2) (collected fractions with retention times from 13.0 to 13.8 min) were combined and lyophilized. The final yield of peak ID: AMR100489-23-F2-P2 (lot number #IN-SDV-D-168-2) was 1.4 g.

[0509] Table 3. Preparative HPLC Method Conditions for Primary Treatment of Lot No. #AMR100489-23-F2

[0510]

[0511] Approximately 109 g of lot #LHGE-180125 was processed using the primary preparative HPLC method described in Table 4. The fraction with peak ID: LHGE-180125-P6 (lot #IN-RAS-A-24-6) (collected fractions with retention times from 25.5 to 29.0 min) was combined and lyophilized. The final yield of peak ID: LHGE-180125-P6 (lot #IN-RAS-A-24-6) was 5.5 g.

[0512] Table 4. Preparative HPLC Method Conditions for Primary Treatment of Lot No. #LHGE-180125

[0513]

[0514] Secondary processing: Approximately 1.4 g of peak ID: AMR100489-23-F2-P2 (lot number #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 (used for collection within a retention time range of 26.0 to 27.0 min) collected from the secondary processing of #AMR100489-23-F2-P2 were combined and lyophilized for separation, as described in section 3.8. The final yield of peak ID: AMR100489-23-F2-P2-G (lot number #IN-VVP-K-138-7) was 34 mg with a purity of 89.0% (area %).

[0515] Table 5. Preparation method conditions for secondary treatment of peak ID: AMR100489-23-F2-P2 (lot number #IN-SDV-D-168-2)

[0516]

[0517] Approximately 5.5 g of peak ID: LHGE-180125-P6 (lot number #IN-RAS-A-24-6) was processed using the preparative HPLC method conditions described in Table 6. The fraction with peak ID: LHGE-180125-P6-B (the time range for which the fraction was collected was retention time 5.0 to 5.50 min) was combined and lyophilized. The final yield of peak ID: LHGE-180125-P6-B (lot number #IN-RAS-A-87-2) was 800 mg.

[0518] Table 6. Preparation method conditions for secondary processing of peak ID: LHGE-180125-P6 (lot number #IN-RAS-A-24-6)

[0519]

[0520]

[0521] Third-stage processing: Approximately 34 mg of the peak ID: AMR100489-23-F2-P2-G (lot number #IN-VVP-K-138-7) was further processed using the preparative HPLC method conditions described in Table 7. The target fraction had retention times ranging from 6.3 to 6.9 min. The target fractions were combined and lyophilized. The final yield of high-purity peak ID: AMR100489-23-F2-P2-G (lot number #IN-VVP-K-172) was 13 mg with a purity of 95.5% (area%).

[0522] Table 7. Preparation method conditions for the three-stage treatment of peak ID: AMR100489-23-F2-P2-G (lot number #IN-VVP-K-137-8)

[0523]

[0524]

[0525] The peak ID: LHGE-180125-P6-B (lot number #IN-RAS-A-87-2) at approximately 800 was processed using the preparative HPLC method conditions described in Table 8. The target fraction had a retention time of 23.0 to 25.0 min. The target fractions were combined and lyophilized. The final yield of peak ID: LHGE-180125-P6-B7 (lot number #IN-RAS-A-106-7) was 15 mg.

[0526] Table 8. Level III preparation method conditions for peak ID: LHGE-180125-P6-B (lot number #IN-RAS-A-87-2)

[0527]

[0528]

[0529] Level IV processing: Approximately 15 mg of peak ID: LHGE-180125-P6-B-7 (lot number #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 retention times from 22.0 to 22.8 min. The target fractions were combined and lyophilized. The final yield of peak ID: LHGE-180125-P6-B7-C (lot number #IN-RAS-A-119-3) was 0.9 mg with a purity of 81.3% (area%).

[0530] The fractions collected from the preparative process were combined and freeze-dried using a Labconco freeze dryer (the collector temperature was maintained at -44°C under vacuum).

[0531] Table 9. Preparation method conditions for level four peak ID: LHGE-180125-P6-B-7 (lot number #IN-RAS-A-106-7)

[0532]

[0533]

[0534] MS and MS / MS.MS and MS / MS data were generated using a Waters QTof Micro mass spectrometer equipped with an electrospray ionization source. Samples were analyzed by negative ESI. The sample (approximately 0.2 mg) was diluted to approximately 0.2 mg / mL with 50:50 ACN:H2O and introduced via direct perfusion.

[0535] ESI-TOF mass spectra obtained by perfusing the CC-00520 sample showed [MH] at m / z 1417.6843. - Ions. [MH] - The mass of the ion and the expected molecular formula C 65 H 110 O 33 Very consistent (for C) 65 H 109 O 33 The calculated value is 1417.6851 (error: -0.6ppm).

[0536] NMR spectroscopy. Obtain a series of NMR experiments, including 1 H NMR, 13 C NMR, 1 H- 1 H COSY, HSQC-DEPT, HMBC, ROESY, and 1D TOCSY are used to allow the specification of CC-00520.

[0537] Table 10. 1 H and 13 C NMR (500 and 125 MHz, CD3OD), designated as CC-00520 aglycone.

[0538]

[0539]

[0540] The terms can be interchanged.

[0541] Table 2. 1 H and 13 C10 NMR (500 and 125 MHz, CD3OD), CC-00520, designated as C-3 glycoside.

[0542]

[0543]

[0544] λThere are four carbon resonances in the range of 75.1–75.3 ppm (75.09 ppm, 75.17 ppm, 75.29 ppm and 75.32 ppm), so the chemical shift cannot be clearly specified.

[0545] § There are seven carbon resonances in the range of 77.7–78.1 ppm (77.67 ppm, 77.88 ppm, 77.91 ppm, 78.06 ppm, and 78.14 ppm; two additional carbon resonances overlap in this region), so the chemical shift cannot be definitively specified.

[0546] There are three carbon resonances at 71.6 ppm (71.55 ppm and 71.60 ppm; the two carbons overlap at 71.60 ppm), so the chemical shift cannot be definitively specified.

[0547] There are two carbon resonances at 62.7 ppm (62.71 ppm and 62.73 ppm), so the chemical shift cannot be definitively specified.

[0548] CC-00520 was identified as mogroside-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 in that additional xylose units are attached to GlcIV via 1→4 glycosidic bonds.

[0549] Example 9: Purification and Characterization of CC-00539

[0550] CC-00539 (mogroside-3-O-{[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}-24-O-{[α-L-rhamnosyl-(1→2)]-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranoside}) was isolated from a Luo Han Guo extract containing 90% mogroside V. The isolation of this compound involved multiple treatments using preparative HPLC to obtain 12 mg (70% purity) of CC-00539. NMR data indicated that the sample contained approximately 70% CC-00539 and approximately 30% mogroside V.

[0551] 1 H, 13 C 1 H- 1 H COSY、1 H- 13 C HSQC-DEPT and 1 H- 13 C HMBC data indicate the presence of a central triterpenoid core and five sugar units in the structure. Based on 1 H, 13 C 1 H- 1 H COSY, 1D-TOCSY, 1 H- 13 C HSQC-DEPT and 1 H- 13 C HMBC data determines the complete structure. 1 In H NMR, in addition to the eight methyl protons observed for the central triterpene nucleus, in δ H The additional methyl group observed at 1.21 (d, J = 6.2 Hz) indicates the presence of a rhamnose unit in the structure. 1D and 2D NMR data indicate that the remaining four sugar units are glucose. δ¹⁸ NMR values ​​were obtained using different mixing times (20–140 ms). H 1D-TOCSY data of the terminal matrix at 5.31 confirms that at δ H The methyl proton at 1.21 belongs to the same spin system, and therefore δ H The terminal proton at 5.31 is designated as rhamnose H-1. In δ H The terminal protons of rhamnose observed at position 5.31 show a similarity to Glc I C-2 HMBC correlation (δ) C 77.4). It was also observed from Glc... I H-2(δ H 3.49) to the terminal carbon of rhamnose (δ C The reciprocal HMBC correlation of 102.0 indicates that rhamnose and Glc I The 1→2 bond between them. This was obtained using a 500MHz NMR instrument equipped with a 2.5mm reverse probe. 1 In the 1H NMR spectrum, a broad singlet was observed at the terminal protons of rhamnose, indicating the α-configuration of rhamnose. (The text abruptly ends here, likely due to an incomplete sentence or missing information.) 13 Before performing C10 NMR spectroscopy, when using a 500MHz NMR instrument equipped with a 5mm broadband probe to obtain... 1 During 1H NMR spectroscopy, the spectra were better resolved and the rhamnose terminal protons were observed as broad doublets with a coupling value of 1.1 Hz, confirming the presence of the α-configuration of rhamnose. This will be crucial... 1 H- 1 H COSY and 1 H- 13C HMBC correlation is used to identify sugar bonds in the structure.

[0552] Mass spectrometry analysis of CC-00539 in negative polarity mode by electrospray ionization time-of-flight (ESI-TOF) mass spectrometry showed [MH] at m / z 1269.6537. - Ions. [MH] - The mass of the ion and the expected molecular formula C 60 H 102 O 28 Very consistent (for C) 60 H 101 O 28 The calculated value is 1269.6479 (error: 4.6 ppm).

[0553] Example 10: Preparation, purification and characterization of CC-00540

[0554] CC-00540 was isolated from the reaction product of mogroside IIIE with α-D-xylosylfluorine and α-xylosidase. CC-00540 was purified using a multi-round preparative HPLC method to provide approximately 3.7 mg with a purity of 95.4% (HPLC area %). A series of 1D and 2D experiments were conducted on this sample: 1 H, 13 C 1 H- 1 H COSY、 1 H- 13 C HSQC-DEPT, 1 H- 13 CHSQC-TOCSY, and 1 H- 13 C HMBC was used to elucidate the structure as mogroside-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]. Spectroscopic data revealed a central triterpenoid core and four sugar units: two glucose units and two xylose units. The bonds between the sugar units and their attachment to the central triterpenoid core were determined based on COSY, TOCSY, and HSQC-DEPT data. The two glucose units exist as β-terminal isomers, while the two xylose units (… 3 J = 3.1 Hz) exists in the α configuration.

[0555] Table 1. 1 H NMR (500.13MHz, CD3OD) and 13 C NMR (125MHz) specified CC-00540

[0556]

[0557]

[0558] Example 11: Preparation, purification and characterization of CC-00541

[0559] CC-00541 was isolated from the reaction product of mogroside IIIE with α-D-xylosylfluorine and α-xylosidase. CC-00541 was purified using multiple rounds of preparative HPLC to provide approximately 11.1 mg of CC-00541 with a purity of 83.8% (HPLC area %), as well as CC-00540 as a minor compound. A series of 1D and 2D experiments were conducted on this sample: 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 used to elucidate the structure as mogroside-3-O-[β-D-glucopyranoside]-24-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]. Spectroscopic data revealed the presence of a central triterpenoid core and three sugar units: two glucose units and one xylose unit. The bonds between the sugar units and their attachment to the central triterpenoid core were determined based on COSY, TOCSY, and HSQC-DEPT data. The two glucose units exist as β-terminal isomers, while the remaining xylose unit ( 3 J = 3.6 Hz exists in the α configuration.

[0560] Table 1. 1 H NMR (500.13MHz, CD3OD) and 13 C NMR (125MHz) specified CC-00541

[0561]

[0562]

[0563] Example 12: Preparation, purification and characterization of CC-00542

[0564] CC-0542 was isolated from the reaction product of mogroside IIIE with α-D-xylosylfluoride and α-xylosidase. CC-00542 was purified using multiple rounds of preparative HPLC to provide approximately 6.8 mg with a purity of 84% (HPLC area %). A series of 1D and 2D experiments were conducted on this sample: 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 used to elucidate the structure as mogroside-3-O-[α-D-xylopyranosyl-(1→6)-β-D-glucopyranoside]-24-O-β-D-glucopyranoside. Spectroscopic data indicated the presence of a central triterpenoid core and three sugar units: two glucose units and one xylose unit. The bonds between the sugar units and their attachment to the central triterpenoid core were determined based on COSY, TOCSY, and HSQC-DEPT data. The two glucose units existed as β-terminal isomers, while the remaining xylose unit ( 3 J = 3.6 Hz exists in the α configuration.

[0565] Table 1. 1 H NMR (500.13MHz, CD3OD) and 13 C NMR (125MHz) is specified as CC-00542.

[0566]

[0567]

[0568] Example 13: Preparation, purification and characterization of CC-00550

[0569]

[0570] 3-O-β-D-glucopyranosylmogromellose 24-O-α-L-rhamnosyl-(1→2)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside (4). At room temperature under N2 atmosphere, 0.5M solution of NaOMe in MeOH (0.52mL, 0.2625mmol) was added dropwise to a solution of 3 (613.8mg, 0.2625mmol) in dry THF (11mL) and dry MeOH (11mL). The mixture was stirred at the same temperature for 18h, neutralized with 10% AcOH aqueous solution and concentrated under reduced pressure. The remaining white solid was passed through a preparative HPLC column (Phenomenex Luna C18(2), 21.2×250mm, 5μm particles, 25% CH3CN in H2O, 20mL / min, t R =10.2 min) purification. The combined fractions were concentrated under reduced pressure and the residue was dried under vacuum to give title compound 4 (225.5 mg, 77%) as a white powder. R f =0.68 (silica gel, CH2Cl2 / MeOH / H2O = 5:4:1). 11H NMR (500 MHz, pyridine-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 = 5.7 Hz, 1H), 5.02 (d, J = 7.8 Hz, 1H), 4.92 (d, J = 7.8 Hz, 2H), 4.87 - 4.82 (m, 1H), 4.82 - 4.74 (m, 2H), 4.62 (dd, J = 9.2, 3.3 Hz, 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 = 6.2 Hz, 3H), 1.73 - 1.65 (m, 3H), 1.63 (d, J = 7.5 Hz, 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 = 4.3 Hz, 3H), 1.05 - 0.96 (m, 1H), 0.89 (s, 3H), 0.81 (s, 3H). 13C10 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.

[0571] Example 14: Preparation, purification and characterization of CC-00551

[0572] CC-00551 was isolated from a Luo Han Guo extract containing 90% mogroside V. 1D and 2D NMR and MS spectral analysis of CC-00551 allowed for the complete specification of its structure as mogroside-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 specified by a combination of 1H, 13C, 1H-1H COSY, 1H-13C HSQC-DEPT, and 1H-13CHMBC data, while a series of 1D-TOCSY experiments combined with 1D and 2D NMR data were used for the specification of sugar units and bonds. Evaluation of the NMR data concluded that this compound contains four glucose units and one xylose unit. Xylose units are linked to GlcI via 1→6 sugar bonds.

[0573] ESI-TOF mass spectra obtained from samples perfused with CC-00551 showed [MH]- ions at m / z 1255.6377. The mass of the [MH]- ion is consistent with the expected molecular formula C. 59 H 100 O 28 Very consistent (for C) 59 H 99 O 28 The calculated value is 1255.6323 (error: 4.3 ppm). MS data confirms the nominal mass of 1256 Daltons, with the molecular formula C. 59 H 100 O 28The ions observed at m / z 1291.5961 and 1353.5873 are most likely attributed to [M-H+HCl]- and [M-H+H3PO4]-, respectively. MS / MS spectroscopy (fragmentation of the [MH]- ion at m / z 1255.6) indicates the loss of a xylose unit at m / z 1123.5693, followed by the successive loss of four glucose units at m / z 961.5647, 799.5051, 637.4304, and 475.3798, suggesting the presence of one xylose unit and four glucose units in the structure.

[0574] Table 1. 1 H and 13 C NMR (500 and 125 MHz, CD3OD), designated as CC-00551 aglycone.

[0575]

[0576]

[0577] The terms can be interchanged.

[0578] Table 2. 1 H and 13 C10 NMR (500 and 125 MHz, CD3OD), designation of CC-00551C-3 glycoside.

[0579] Location <![CDATA[ 13 C]]> <![CDATA[ 1 H]]> <![CDATA[Glc IV -1]]> 106.4 4.28d(7.8) <![CDATA[Glc IV -2]]> 75.6 3.19m <![CDATA[Glc IV -3]]> <![CDATA[77.9-78.2 § ]]> Approximately 3.31m <![CDATA[Glc IV -4]]> <![CDATA[71.6-71.9 λ ]]> Approximately 3.30m <![CDATA[Glc IV -5]]> 77.2 3.40m <![CDATA[Glc IV -6]]> 69.8 <![CDATA[3.80m,4.05brdd(10.3 Ψ )]]> <![CDATA[Glc V -1]]> 104.8 4.42d(7.8) <![CDATA[Glc V -2]]> 75.1 or 75.2 3.18m <![CDATA[Glc V -3]]> <![CDATA[77.9-78.2 § ]]> 3.35m <![CDATA[Glc V -4]]> <![CDATA[71.6-71.9 λ ]]> 3.28m <![CDATA[Glc V -5]]> <![CDATA[77.9-78.2 § ]]> Approximately 3.26m <![CDATA[Glc V -6]]> 62.7 3.66m, 3.85m

[0580] § Five carbon resonances (77.93ppm, 77.98ppm, 78.06ppm, 78.14ppm and 78.19ppm) in the range of 77.9-78.2ppm, therefore the chemical shift cannot be clearly specified.

[0581] λ There are three carbon resonances (71.59 ppm, 71.65 ppm and 71.88 ppm) in the range of 71.6–71.9 ppm, so the chemical shift cannot be clearly specified.

[0582] Ψ The signal was not fully resolved, so smaller couplings could not be identified.

[0583] Table 3. 1 H and 13 C10 NMR (500 and 125 MHz, CD3OD), designation of CC-00551C-24 glycoside.

[0584] Location <![CDATA[ 13 C]]> <![CDATA[ 1 H]]> <![CDATA[Glc I -1]]> 104.1 4.43d(7.3) <![CDATA[Glc I -2]]> 81.3 3.61m <![CDATA[Glc I -3]]> 78.8 3.58m <![CDATA[Glc I -4]]> <![CDATA[71.6-71.9 λ ]]> Approximately 3.28m <![CDATA[Glc I -5]]> 76.5 3.51m <![CDATA[Glc I -6]]> 70.5 <![CDATA[3.51m,4.21 € ]]> <![CDATA[Glc III -1]]> 104.5 4.77d(7.8) <![CDATA[Glc III -2]]> 75.7 3.27m <![CDATA[Glc III -3]]> <![CDATA[77.9-78.2 § ]]> 3.36m <![CDATA[Glc III -4]]> 72.3 3.21m <![CDATA[Glc III -5]]> <![CDATA[77.9-78.2 § ]]> Approximately 3.27m <![CDATA[Glc III -6]]> 63.5 3.63m, 3.86m Xyl-1 105.4 4.22d(7.4) Xyl-2 75.1 or 75.2 3.19m Xyl-3 77.5 3.31m Xyl-4 71.2 3.47m Xyl-5 66.9 3.18m, 3.84m

[0585] § Five carbon resonances (77.93ppm, 77.98ppm, 78.06ppm, 78.14ppm and 78.19ppm) in the range of 77.9-78.2ppm, therefore the chemical shift cannot be clearly specified.

[0586] λ There are three carbon resonances (71.59 ppm, 71.65 ppm and 71.88 ppm) in the range of 71.6–71.9 ppm, so the chemical shift cannot be clearly specified.

[0587] € Resonance that partially overlaps with the xylose terminal protons. Therefore, diversity cannot be specified.

[0588] Example 15: Sensory Analysis

[0589] The following samples were tested in water at 400 ppm at 4°C. Sample volumes were limited. Therefore, the number of expert panel members for each sample was 1–10 and the test volume was 2–10 mL. The sweetness of each sample was compared with a sucrose reference.

[0590] compound Sweetness (SE) CC-00489 >9SE CC-00491 >10 CC-00500 >5 CC-00518 >9 CC-00520 >7 CC-00539 >3 CC-00540 >2 CC-00541 >1 CC-00542 >2

[0591] Example 16: Preparation of CC-00497

[0592] CC-00497 was prepared by biotransformation of isomogroside V. 250 mg of isomogroside V and 10 mg of β-galactosidase G5160 were stirred in 3.1 mL of pH 5 sodium acetate buffer for 3 days and heated for 30 minutes at 37 °C. The crude mixture was directly purified by preparative HPLC to provide 2.8 mg of CC-00497. The structure was confirmed by 1D and 2D NMR analysis.

[0593] Table 1. 1 H and 13 C NMR (500 and 125 MHz, CD3OD), designation of CC-00497 aglycone.

[0594]

[0595]

[0596]

[0597] The terms can be interchanged. § There are two carbon resonances at 37.3 ppm (37.25 ppm and 37.31 ppm), so the chemical shift cannot be definitively specified.￥ There are two carbon resonances at 26.2 ppm (26.20 ppm and 26.23 ppm), so the chemical shift cannot be definitively specified. Partially overlapping methyl resonances.

[0598] Table 2. 1 H and 13 C10 NMR (500 and 125 MHz, CD3OD), designation of CC-00497C-3 glycoside.

[0599]

[0600] € There are four carbon resonances in the range of 75.1–75.5 ppm (75.12 ppm, 75.15 ppm, 75.18 ppm and 75.54 ppm), so the chemical shift cannot be clearly specified.

[0601] § There are four carbon resonances in the range of 77.9–78.1 ppm (77.89 ppm, 77.95 ppm, 78.04 ppm, and 78.08 ppm), so the chemical shift cannot be definitively specified.

[0602] λ There are two carbon resonances at 71.6 ppm (71.57 ppm and 71.62 ppm), so the chemical shift cannot be definitively specified.

[0603] Table 3. 1 H and 13 C10 NMR (500 and 125 MHz, CD3OD), designation of CC-00497C-24 glycoside.

[0604]

[0605] € There are four carbon resonances in the range of 75.1–75.5 ppm (75.12 ppm, 75.15 ppm, 75.18 ppm and 75.54 ppm), so the chemical shift cannot be clearly specified.

[0606] § There are four carbon resonances in the range of 77.9–78.1 ppm (77.89 ppm, 77.95 ppm, 78.04 ppm, and 78.08 ppm), so the chemical shift cannot be definitively specified.

[0607] ￡ Due to Glc at 4.29 ppm I H-1 and Glc at 4.28 ppm IVThe partial overlap and impurities of H-1 make it impossible to explicitly specify the coupling constant.

Claims

1. A mogroside, selected from the following structures: ， CC-00489 ， CC-00491 ， CC-00500 ,and CC-00518 CC-00520.

2. A composition comprising mogroside as claimed in claim 1 and at least one other substance, wherein the composition comprises at least 5% by weight mogroside as claimed in claim 1.

3. A consumer product comprising mogroside as described in claim 1 or a composition as described in claim 2.

4. The consumer product of claim 3, wherein the consumer product is a beverage.

5. The consumer product of claim 4, wherein the beverage contains mogroside at a concentration of 10 ppm to 1,000 ppm or 1 ppm to 100 ppm.

6. The consumer product of claim 4, wherein the beverage further comprises at least one additional sweetener selected from the group consisting of carbohydrate sweeteners, high-efficiency sweeteners, synthetic sweeteners, and combinations thereof.

7. The consumer product of claim 4, wherein the beverage further comprises at least one additional sweetener selected from the group consisting of: rebaudin A, rebaudin M, rebaudin D, mogroside V, symmonin I, and mogroside-3-O-[β-D-glucopyranoside]-24-O-{[β-D-glucopyranoside-(1→2)]-[α-D-glucopyranoside-(1→6)]-β-D-glucopyranoside}.

8. The consumer product of claim 4, wherein the beverage is carbonated.

9. The consumer product of claim 4, wherein the beverage is non-carbonated.

10. The consumer product of claim 4, wherein the beverage comprises an organic acid salt.

11. The consumer product of claim 3, wherein the consumer product is a beverage having a serving size of up to 40 calories per 8 ounces.

12. The consumer product of claim 3, wherein the consumer product is a beverage having a serving size of less than 5 calories per 8 ounces.

13. The consumer product of claim 6, wherein the additional sweetener is a carbohydrate sweetener, and the carbohydrate sweetener is a rare sugar sweetener.

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