dihydrochalcones from kandelia candel

Abstract

Provided herein are methods of isolating dihydrochalcone compounds having Formula (I) from B. rubra. Also provided are compositions and consumables comprising at least one sweetener and at least one dihydrochalcone compound described herein. Also detailed herein are methods of enhancing the sweetness of a consumable, methods of sweetening a consumable to taste more like sucrose, and methods of preparing a consumable.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Application No. 62 / 749,993, filed October 24, 2019, which is incorporated herein by reference in its entirety. Invention Field

[0003] This instruction relates to a method for isolating dihydrochalcone compounds from Balanophora harlandii and the use of said dihydrochalcone compounds in consumer products. Background of the Invention

[0005] Naturally occurring sugars with calories, such as sucrose, fructose, and glucose, are used to give beverages, food, pharmaceuticals, and oral hygiene / cosmetic products a pleasant taste. In particular, sucrose imparts a flavor that consumers prefer. While sucrose offers superior sweetness, it is disadvantageously high in calories.

[0006] Calorie-free or low-calorie sweeteners have been introduced to meet consumer demand. However, calorie-free and low-calorie sweeteners differ from naturally calorie sugars in ways that frustrate consumers. Based on taste, calorie-free or low-calorie sweeteners exhibit different time characteristic curves, maximum response, flavor characteristic curves, mouthfeel, and / or adaptation behavior compared to sugars. Specifically, calorie-free or low-calorie sweeteners exhibit delayed sweetness onset, persistent sweet aftertaste, bitterness, metallic taste, astringency, cooling taste, and / or a licorice-like flavor. On a source basis, many calorie-free or low-calorie sweeteners are synthetic chemicals. Consumers' expectations for natural calorie-free or low-calorie sweeteners that taste like sucrose remain high.

[0007] Stevia rebaudiana Bertoni is a perennial shrub belonging to the Asteraceae family (Compositae) native to certain regions of South America. Its leaves have been used for centuries in Paraguay and Brazil to sweeten local teas and medicines. The plant is commercially cultivated in Japan, Singapore, Malaysia, South Korea, China, Israel, India, Brazil, Australia, and Paraguay.

[0008] The leaves of the plant contain a mixture of diterpenoid glycosides, in amounts ranging from about 10% to 15% of the total dry weight. These diterpenoid glycosides are about 30 to 450 times sweeter than sugars. Structurally, the diterpenoid glycosides are characterized by a single base structure, steviol, but differ in the presence of carbohydrate residues at the C13 and C19 positions. Typically, based on dry weight, the four main steviol glycosides found in stevia leaves are durqueside A (0.3%), rebaudioside C (0.6%–1.0%), rebaudioside A (3.8%), and steviol glycoside (9.1%). Other glycosides identified in stevia extracts include rebaudiosides B, D, E, and F, steviol disaccharides, and stevia glycosides. Of these, only steviol glycoside and rebaudioside A are commercially available.

[0009] Mogrosides, derived from monk fruit, are the common name for a sweet extract made from the fruit of the monk fruit (Siraitia grosvenorii), a perennial herbaceous vine native to southern China and northern Thailand. Monk fruit extract is nearly 250 times sweeter than sugar and is calorie-free. The sweetness of monk fruit is generally attributed to mogrosides.

[0010] To date, the use of steviol glycosides and mogrosides has been limited by certain undesirable taste characteristics, including licorice, bitterness, astringency, sweet aftertaste, bitter aftertaste, and licorice residue. These taste characteristics become more pronounced at higher concentrations and give consumer products (e.g., beverages) a taste different from sucrose. Furthermore, the maximum sweetness of most steviol glycosides and mogrosides is generally lower than the acceptable sweetness level in conventional beverage formulations for sweetened consumer products (e.g., beverages). Invention Overview

[0012] This course covers dihydrochalcone compounds isolated from red winter jasmine and their use in consumer products such as beverages.

[0013] In one aspect, a method is provided for purifying dihydrochalcone compounds having formula I from *Hemiberlesia lataniae*.

[0014]

[0015] Where R 1 and R 2 Each is independently selected from OH and OR 6 ;R 3 Selected from hydrogen and CH2COOH; R 4 Selected from hydrogen and CH3; and R 6The sugar is optionally substituted. The method comprises (a) providing a crude extract of *Hemiberlesia argyi* plant material; (b) passing the crude extract through a column packed with a polymer adsorption resin and eluting it with an aqueous ethanol solvent containing at least about 75% ethanol to provide a first solution containing at least one dihydrochalcone of formula I; and (c) purifying the first solution containing at least one dihydrochalcone of formula I by reversed-phase chromatography to provide a second solution containing at least one dihydrochalcone of formula I.

[0016] In one aspect, the second solution may then be further processed in one or more steps to provide a single dihydrochalcone of formula Ia selected from CC-00487, CC-00486 and CC-00474, with a purity of at least about 80% by weight on a dry basis.

[0017]

[0018] In another aspect, the second solution can then be further processed in one or more steps to provide a single dihydrochalcone of formula Ib selected from CC-00549, CC-00536 and CC-00537, with a purity of at least about 80% by weight on a dry basis:

[0019]

[0020] In another aspect, a composition comprising at least one sweetener and at least one dihydrochalcone (i.e., a dihydrochalcone compound having formula I, Ia, and Ib) is provided. The at least one sweetener may be any known sweetener and is present in a sweetening amount. In one embodiment, the sweetener is selected from at least one steviol glycoside, at least one mogroside, or a combination thereof.

[0021] In one specific embodiment, the at least one dihydrochalcone compound described herein is present at or below its sweetness identification threshold concentration.

[0022] In one embodiment, the at least one dihydrochalcone compound described herein is present in the composition such that when the composition is added to a consumer product, the sucrose equivalent (SE) of the consumer product is enhanced by at least about 1.2 times compared to the SE of a consumer product in the absence of the at least one dihydrochalcone compound.

[0023] In another embodiment, the at least one dihydrochalcone compound described herein is present in the composition such that when the composition is added to a consumer product, it modifies one or more flavor properties of the sweetener, thereby making the consumer product taste more like a sucrose-sweetened product compared to a consumer product in the absence of the at least one dihydrochalcone compound.

[0024] Consumer products comprising at least one sweetener and at least one dihydrochalcone compound described herein, wherein the sweetener is present in a sweetening amount.

[0025] In one embodiment, the at least one dihydrochalcone compound described herein is present in the consumer product in an amount that effectively enhances the sucrose equivalent (SE) of the consumer product by at least about 1.2 times compared to the SE of the consumer product in the absence of the at least one dihydrochalcone compound.

[0026] In another embodiment, the at least one dihydrochalcone compound described herein is present in the consumer product to effectively modulate one or more flavor properties of the sweetener, thereby making the consumer product taste more like a sucrose-sweetened product compared to a consumer product in the absence of the at least one dihydrochalcone compound.

[0027] In one embodiment, the at least one dihydrochalcone compound described herein is present in a consumer product at a concentration from about 1 ppm to about 50 ppm.

[0028] In one embodiment, the at least one sweetener is present in the consumer product at a concentration of about 50 ppm to about 600 ppm.

[0029] Exemplary consumer products include, but are not limited to, pharmaceutical compositions, edible gels and mixtures, dental compositions, sweets, condiments, chewing gum, cereal compositions, baked goods, dairy products, tabletop sweeteners, beverages, and beverage products.

[0030] In a specific embodiment, the consumer product is a beverage or beverage product. In embodiments where the consumer product is a beverage, the beverage may be a high-calorie beverage, a medium-calorie beverage, a low-calorie beverage, or a zero-calorie beverage.

[0031] In another aspect, a method for enhancing the sweetness of a consumer product is provided, the method comprising (i) providing a consumer product containing at least one sweetener and (ii) adding at least one dihydrochalcone compound described herein to the consumer product, thereby providing a consumer product with enhanced sweetness. The sweetness fraction (SE) of the consumer product containing the at least one dihydrochalcone compound is enhanced by at least about 1.2 times compared to the SE of the consumer product in the absence of the at least one dihydrochalcone compound.

[0032] In another embodiment, a method for enhancing the sweetness of a consumer product is provided, the method comprising (i) providing a consumer product matrix and (ii) adding at least one sweetener and at least one dihydrochalcone compound described herein to the consumer product matrix to provide a consumer product with enhanced sweetness. The at least one dihydrochalcone compound is added to the consumer product matrix in an amount that effectively enhances the sweetness (SE) of the consumer product by at least about 1.2 times compared to the SE of a consumer product in the absence of the at least one dihydrochalcone.

[0033] A method is also provided for making a consumer product taste more like sucrose sweetening, the method comprising (i) providing a consumer product containing at least one sweetener and (ii) adding an amount of at least one dihydrochalcone compound described herein to the consumer product to effectively modulate one or more flavor properties of the sweetener to taste more like sucrose sweetening when compared to a consumer product in the absence of the at least one dihydrochalcone compound.

[0034] In another embodiment, a method is provided to make a consumer product taste more like sucrose sweetening, the method comprising (i) providing a consumer product matrix and (ii) adding at least one sweetener and at least one dihydrochalcone compound described herein to the consumer product matrix. The at least one dihydrochalcone compound is added to the consumer product matrix in an amount that effectively modulates one or more flavor properties of the sweetener, thereby making the consumer product taste more like sucrose sweetening when compared to a consumer product in the absence of the at least one dihydrochalcone compound. Attached Figure Description

[0035] Figure 1 The isolation schemes of CC-00487, CC-00486, CC-00474, CC-00536, CC-00537, and CC-00549 from *Hemiberlesia lataniae* are shown. Detailed Implementation

[0036] I. Definition

[0037] As used herein, the term “consumer product” means a substance that comes into contact with the mouth of a person or animal, including substances that are ingested and subsequently expelled from the mouth and substances that are consumed, eaten, swallowed or otherwise ingested, and that would be healthy to a person or animal when used in a generally acceptable manner.

[0038] As used herein, the term "sweetness enhancer" refers to a compound that, when present in a consumer product (e.g., a beverage) at a concentration at or below the sweetener recognition threshold of the compound (i.e., a concentration at which the compound would not produce any noticeable sweetness in the absence of one or more additional sweeteners), enhances, amplifies, or strengthens the perceived sweetness of the product. The term "sweetness enhancer" is synonymous with the terms "sweetness synergist," "sweetness enhancer," "sweetness amplifier," and "sweetness fortifier."

[0039] As is commonly used herein, the term "sweetness recognition threshold" is the lowest known concentration of a compound that can be perceived as sweet by human taste. Sweetness recognition threshold concentrations are specific to particular compounds and can vary based on temperature, matrix, composition, and / or flavor system.

[0040] As used herein, the term "flavor modifier" refers to a compound that actively influences the perception of non-sucrose sweeteners in a consumer product (e.g., a beverage) in a way that makes the consumer taste a more sucrose-sweetened beverage. For example, flavor modifiers can mask certain negative taste characteristics of non-sucrose sweeteners, such as bitterness, sourness, astringency, saltiness, and metallic notes. In another instance, they can improve mouthfeel. In yet another instance, they can reduce sweetness retention. In yet another instance, they can increase sweetness onset. In yet another instance, they can improve sweetness onset. In still another instance, they can improve bitterness retention.

[0041] II. Methods for separating from *Smilax china*

[0042] A method is provided for purifying a dihydrochalcone compound having formula I from *Hemiberlesia oleracea*, the method comprising:

[0043] (a) Provide crude extracts of the plant material of Red Winter Snakehead;

[0044] (b) Passing the crude extract through a column packed with a polymer adsorption resin and eluting with aqueous ethanol containing at least about 75% ethanol to provide a solution containing at least one dihydrochalcone compound of formula I; and

[0045] (c) Purify the solution containing at least one dihydrochalcone compound by reversed-phase chromatography to provide a purified dihydrochalcone compound having Formula I.

[0046] Red winter snakehead is a parasitic plant belonging to the genus *Symplocos*, typically distributed in temperate and tropical Asia, Africa, and the Pacific region. All species in the genus *Symplocos* act as parasites on the roots of host plants. *Symplocos* species possess underground tubers, an inconspicuous flowering system, colorful scaly leaves, and small flowers.

[0047] All or part of the *Sclerotium rubrum* plant can be used in the method of the present invention. In one embodiment, the *Sclerotium rubrum* plant material is partially dried prior to extraction. In another embodiment, the *Sclerotium rubrum* plant material is completely dried prior to extraction. In some embodiments, the plant material is optionally ground prior to extraction.

[0048] Plant materials can be extracted using any suitable extraction process, such as continuous or intermittent reflux extraction, supercritical fluid extraction, enzyme-assisted extraction, microbial-assisted extraction, ultrasound-assisted extraction, microwave-assisted extraction, etc. These methods can be deployed at any scale.

[0049] The solvent used for extraction can be any suitable solvent, such as polar organic solvents (degassed, vacuum-treated, pressurized, or distilled), nonpolar organic solvents, water (degassed, vacuum-treated, pressurized, deionized, distilled, carbon-treated, or reverse osmosis), or mixtures thereof. In one specific embodiment, the solvent comprises water and one or more alcohols. In another specific embodiment, the solvent comprises water and an alcohol selected from methanol, ethanol, n-propanol, 2-propanol, 1-butanol, 2-butanol, and mixtures thereof. In a more specific embodiment, the solvent contains water and ethanol.

[0050] The amount of alcohol in the solvent can vary from about 10% to about 100%, such as any range like about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or between. In one specific embodiment, the ethanol comprises about 70% to about 90% solvent, with the remainder being water.

[0051] In some embodiments, the extraction process occurs at high temperatures, such as from about 50°C to about 100°C. In a more specific embodiment, the extraction process occurs at a temperature from about 60°C to about 80°C, or about 70°C.

[0052] Those skilled in the art will understand that the duration of extraction will vary based on the amount of solvent and plant material used. In some embodiments, extraction is performed from about 1 hour to about 24 hours, such as at least about 4 hours, at least about 8 hours, at least about 12 hours, at least about 16 hours, or at least about 20 hours.

[0053] Water and / or any insoluble materials can be removed from the extraction mixture to provide a "crude extract".

[0054] Insoluble plant material can be separated from the extracted mixture, for example, by filtration. Other suitable separation methods include, but are not limited to, gravity filtration, plate and frame filter presses, cross-flow filters, mesh filters, Nutsche filters, belt filters, ceramic filters, membrane filters, microfilters, nanofilters, ultrafiltration, or centrifugation. Optionally, various filter aids, such as diatomaceous earth, bentonite, zeolite, etc., can be used in this process.

[0055] The crude extract is then subjected to one or more chromatographic separations to provide a solution of at least one dihydrochalcone compound described herein.

[0056] The crude extract is passed through one or more columns, such as those packed with polymer adsorption resin. Elution of the one or more columns with aqueous ethanol provides a solution containing at least one dihydrochalcone compound described herein. In one embodiment, 10% ethanol is first applied to the column and the eluent is collected. Typically, the first eluent portion does not contain at least one dihydrochalcone compound described herein. Elution with a solvent containing at least about 75% ethanol (e.g., about 85% ethanol or about 95% ethanol) provides a solution containing at least one dihydrochalcone compound described herein.

[0057] The solution containing at least one of the dihydrochalcone compounds described herein was then subjected to reversed-phase chromatography for further purification. The column can be packed with any material suitable for reversed-phase chromatography, such as an MCI gel column. These fractions were collected and, using LC-MS as a guide, extracted ion chromatograms (EIC or XIC) at quasi-ions or fragments of hesperidin dihydrochalcone monosaccharide glycosides or their analogues (e.g., [MH)). - Further processing (m / z 465) is performed. A gradient of methanol and water is used for elution. In one embodiment, a gradient of 1:4 → 4:1 (methanol:water) is used. The gradient can be continuous or gradual.

[0058] These fractions can be collected based on the percentage of methanol used for elution, for example, 20% methanol, 40% methanol, 50% methanol, 60% methanol, and 80% methanol.

[0059] Extraction of these fractions, pH adjustment, partitioning, and chromatography allow for the separation of the individually purified dihydrochalcone compounds described herein. Figure 1 An exemplary separation scheme is shown.

[0060] As those skilled in the art will understand, the quality of the desired dihydrochalcone compound can be tracked by MS or HPLC-MS throughout the methods used herein, for example, to guide fractionation and purification steps.

[0061] CC-00487 can be separated by diluting a solution of at least one dihydrochalcone compound of formula I with water (e.g., obtained during elution with 20% methanol / 80% water in the reversed-phase chromatography step described above and traced by EIC), acidifying, partitioning, alkalizing, and concentrating. Acidification can be performed with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 1 to about 3, such as a pH of about 1. Partitioning can be performed with any suitable organic solvent (e.g., ethyl acetate). The aqueous portion can be alkalized with any suitable base (e.g., a strong base such as NaOH) to a pH from about 6 to about 8, such as a pH of about 7. The concentrate can then be purified, for example by HPLC or other suitable methods, to provide CC-00487.

[0062] CC-00486 can be separated by diluting a solution of at least one dihydrochalcone of Formula I with water (e.g., obtained during elution with 40% methanol / 60% water in the reversed-phase chromatography step described above and traced by EIC), acidifying, partitioning, and concentrating. Acidification can be performed to a pH from about 1 to about 3, such as a pH like about 1, using any suitable acid (e.g., a strong acid such as HCl). Partitioning can be performed using any suitable organic solvent (e.g., ethyl acetate). The organic fraction can then be concentrated. The concentrate can then be purified, for example by HPLC or other suitable methods, to provide CC-00486.

[0063] CC-00474 can be separated by diluting a solution of at least one dihydrochalcone of Formula I with an aqueous base (e.g., obtained during the 60% methanol / 40% water step of the reversed-phase chromatography described above and traced by EIC), acidifying, partitioning, further acidifying, further partitioning, concentrating, and separating. The aqueous base dilution can be, for example, with an aqueous solution of NaOH. Acidification can be performed with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 8 to about 11, such as a pH of about 10. Partitioning can be performed with any suitable organic solvent (e.g., n-butanol). The aqueous fraction of the partition can be further acidified with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 4 to about 6, such as a pH of about 5. The resulting acidified aqueous solution can be further partitioned with any suitable organic solvent (e.g., n-butanol). The n-butanol fraction can then be concentrated. Concentration can be performed, for example, by HPLC purification of the concentrate to provide CC-00474.

[0064] CC-00549 can be separated by further column chromatography (e.g., HPLC) of the material obtained from the above reversed-phase chromatography step (e.g., during the 50% methanol / 50% water phase of the above reversed-phase chromatography step and followed by EIC).

[0065] Hesperidin dihydrochalcone-4'-β-D-glucoside (HDCG) can be separated by diluting a solution of at least one dihydrochalcone of Formula I with an aqueous base (e.g., obtained during the 60% methanol / 40% water step of the reversed-phase chromatography described above and tracked by EIC), acidifying, partitioning, further acidifying, further partitioning, concentrating, and separating. The aqueous base dilution can be, for example, with an aqueous solution of NaOH. Acidification can be performed with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 8 to about 11, such as a pH of about 10. Partitioning can be performed with any suitable organic solvent (e.g., n-butanol). The aqueous fraction of the partition can be further acidified with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 4 to about 6, such as a pH of about 5. The resulting acidified aqueous solution can be further partitioned with any suitable organic solvent (e.g., n-butanol). The n-butanol fraction can then be concentrated. The concentrate can be purified by, for example, HPLC chromatography to provide HDCG.

[0066] CC-00536 can be separated by diluting a solution of at least one dihydrochalcone compound of Formula I with an aqueous base (e.g., obtained during the 60% methanol / 40% water step of the reversed-phase chromatography described above and traced by EIC), acidifying, partitioning, further acidifying, further partitioning, concentrating, and separating. The aqueous base dilution can be, for example, with an aqueous solution of NaOH. Acidification can be performed with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 8 to about 11, such as about 10. Partitioning can be performed with any suitable organic solvent (e.g., n-butanol). The aqueous fraction of the partition can be further acidified with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 4 to about 6, such as about 5. The resulting acidified aqueous solution can be further partitioned with any suitable organic solvent (e.g., n-butanol). The n-butanol fraction can then be concentrated. The concentrate can be purified by, for example, HPLC chromatography to provide CC-00536.

[0067] CC-00537 can be separated by diluting a solution of at least one dihydrochalcone compound of Formula I with an aqueous base (e.g., obtained during the 60% methanol / 40% water step of the reversed-phase chromatography described above and traced by EIC), acidifying, partitioning, further acidifying, further partitioning, concentrating, and separating. The aqueous base dilution can be, for example, with an aqueous solution of NaOH. Acidification can be performed with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 8 to about 11, such as about 10. Partitioning can be performed with any suitable organic solvent (e.g., n-butanol). The aqueous fraction of the partition can be further acidified with any suitable acid (e.g., a strong acid such as HCl) to a pH from about 4 to about 6, such as about 5. The resulting acidified aqueous solution can be further partitioned with any suitable organic solvent (e.g., n-butanol). The n-butanol fraction can then be concentrated. The concentrate can be purified by, for example, HPLC chromatography to provide CC-00537.

[0068] The method described herein provides a dihydrochalcone compound having a purity of at least about 80% on a dry basis by weight, such as at least about 85%, at least about 90%, at least about 95%, at least about 97%, or at least about 99%. Those skilled in the art will understand that additional purification steps, such as chromatographic separation on various types of columns, can be performed to achieve the aforementioned purity.

[0069] II. Enhancers / Flavor Modifiers

[0070] In one embodiment, the dihydrochalcone compound having Formula I is a sweetness enhancer. In another embodiment, the dihydrochalcone compound having Formula I is a flavor modifier, with or without simultaneously acting as a sweetness enhancer. That is, in some embodiments, the dihydrochalcone compound enhances the sweetness of the sweetener and modulates one or more flavor properties of the sweetener. In other embodiments, the dihydrochalcone compound enhances the sweetness of the sweetener without modulating one or more flavor properties of the sweetener. In still other embodiments, the dihydrochalcone compound modulates one or more flavor properties of the sweetener without enhancing the sweetness of the sweetener.

[0071] In one embodiment, a dihydrochalcone compound having formula I can be used as a sweetness enhancer and / or flavor modifier:

[0072]

[0073] Where R 1 and R 2 Each is independently selected from OH and OR 6 ;R 3 Selected from hydrogen and CH2COOH; R 4 Selected from hydrogen and CH3; and R 6It is an optional sugar substitute.

[0074] As used herein, “sugar” refers to monosaccharides, disaccharides, oligosaccharides, and polysaccharides. A sugar contains at least one carbohydrate. Exemplary carbohydrates include, but are not limited to, sucrose, glyceraldehyde, dihydroxyacetone, erythrose, threose, erythulose, arabinose, lysose, ribose, xylose, ribulose, xylulose, allose, arabinose, galactose, glucose, gulose, idulose, mannose, tarose, fructose, allulose, sorbose, tagatose, mannohepulose, sedohepulose, octanose, fucose, rhamnose, arabinose, menobiose, and sialic acid.

[0075] The sugar can be D- or L-. In one embodiment, the sugar is D-glucose.

[0076] Furthermore, the sugar can be in either the α-configuration or the β-configuration. In one embodiment, the sugar is β-D-glucose.

[0077] In some embodiments, the sugar is substituted. As used herein, “substitution” with respect to sugar means substitution on one or more oxygen atoms at the hydroxyl (-OH) position. Exemplary substituents include, but are not limited to, galloyl, vanillyl, feruloyl, any other mono-, di-, tri-hydroxybenzoyl, and methoxybenzoyl.

[0078] In one specific embodiment, the dihydrochalcone compound is a compound having formula Ia:

[0079]

[0080] In another embodiment, the dihydrochalcone compound is a compound having formula Ib:

[0081]

[0082] The dihydrochalcone compounds described herein may be provided in pure form or as part of a mixture. The mixture may be an extract prepared from a plant or part of a plant, as described above, or may be commercially available.

[0083] In one embodiment, the dihydrochalcone compound described herein constitutes at least about 50% of the mixture by weight, such as at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95%. In a more specific embodiment, the dihydrochalcone compound described herein constitutes at least about 96%, at least about 97%, at least about 98%, or at least about 99% of the mixture by weight.

[0084] Mixtures of at least two of the dihydrochalcone compounds described herein, such as at least three dihydrochalcone compounds, are also considered. The at least two dihydrochalcone compounds described herein may be provided independently in pure form or as part of a mixture. In one embodiment, the at least two dihydrochalcone compounds described herein constitute at least about 50% of the mixture by weight, such as at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 95%. In a more specific embodiment, the at least two dihydrochalcone compounds described herein constitute at least about 96%, at least about 97%, at least about 98%, or at least about 99% of the mixture by weight.

[0085] III. Sweeteners

[0086] A dihydrochalcone compound having Formula I enhances and / or modifies the flavor of at least one sweetener. The at least one sweetener can be any known sweetener, such as a natural sweetener, a natural high-efficiency sweetener, or a synthetic sweetener.

[0087] The at least one sweetener is present in a sweetening amount. 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).

[0088] As used herein, the phrase "natural high-efficiency sweetener" refers to any sweetener that is naturally found in nature and has a sweetening potency greater than that of sucrose, fructose, or glucose, while having fewer calories. Natural high-efficiency sweeteners may be provided as pure compounds or alternatively as part of extracts. As used herein, the phrase "synthetic sweetener" refers to any composition that is not naturally found in nature and has a sweetening potency greater than that of sucrose, fructose, or glucose, while having fewer calories.

[0089] In other embodiments, the at least one sweetener is a carbohydrate sweetener. Suitable carbohydrate sweeteners are selected from, but not limited to, the group consisting of: sucrose, glyceraldehyde, dihydroxyacetone, erythrose, threose, erythritol, arabinose, lysose, ribose, xylose, ribulose, xylulose, allose, azoose, galactose, glucose, gulose, idole, mannose, tarose, fructose, allulose, sorbose, tagatose, mannoheptulose, sedoheptulose, octanulose, fucose, rhamnose, arabinose, mesobiose, sialic acid, and combinations thereof.

[0090] In other embodiments, the at least one sweetener does not contain a carbohydrate sweetener.

[0091] In another embodiment, the additional sweetener is a rare sugar selected from sorbitol, lysoose, ribulose, xylose, xylulose, D-allose, L-ribose, D-tagatose, L-glucose, L-fucose, L-arabinose, malnobiose, and combinations thereof.

[0092] Other sweeteners include symmonoside I, monoglin and its salts (monatin SS, RR, RS, SR), curculin, glycyrrhizic acid and its salts, thaumatin, monellin, mabinlin, brazzein, hernandulcin, lecithin, phlorizin, phlorizin, trifolin, baiyunoside, osladin, polypodoside A, and pectin. Erocaryoside A, pectin B, mukurozioside, phlomisoside I, periandrin I, abrusoside A, stevioside and cyclocaryoside I, sugar alcohols such as erythritol, sucralose, acetylsupan potassium, 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.

[0093] 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.

[0094] In other embodiments, the sweetener comprises at least one steviol glycoside, wherein the at least one steviol glycoside is present in a sweetening amount. The steviol glycoside may be natural, synthetic, or a combination of natural and synthetic.

[0095] Steviosides may be provided in pure form or as part of a mixture (i.e., a steviol glycoside blend). Exemplary steviol glycosides include, but are not limited to, rebaudioside M, rebaudioside D, rebaudioside A, rebaudioside N, rebaudioside O, rebaudioside E, steviol monosaccharide glycoside, steviol disaccharide glycoside, stevioside, durcuroside B, durcuroside A, rebaudioside B, rebaudioside G, steviol glycoside, rebaudioside C, rebaudioside F, rebaudioside I, rebaudioside H, rebaudioside L, rebaudioside K, rebaudioside J, rebaudioside M2, rebaudioside D2, rebaudioside S, rebaudioside T, rebaudioside U, rebaudioside V, rebaudioside W, rebaudioside Z1, rebaudioside Z2, rebaudioside IX, enzymatically glycosylated steviol glycosides, and combinations thereof.

[0096] In some embodiments, the steviol glycoside blend 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% steviol glycoside.

[0097] In an exemplary embodiment, the steviol glycoside blend comprises at least about 50% by weight, such as from about 50% to about 90%, from about 50% to about 80%, from about 50% to about 70%, from about 50% to about 60%, from about 60% to about 90%, from about 60% to about 80%, from about 60% to about 70%, from about 70% to about 90%, from about 70% to about 80%, and from about 80% to about 90%.

[0098] In one embodiment, the sweetener is a steviol glycoside blend containing a sweetening amount of rebaudioside M. In one embodiment, the steviol glycoside blend contains 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% rebaudioside M.

[0099] In another embodiment, the sweetener is a steviol glycoside blend containing a sweetening amount of rebaudioside A. In one embodiment, the steviol glycoside blend contains 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% rebaudioside A.

[0100] In yet another embodiment, the sweetener is a steviol glycoside blend containing a sweetening amount of rebaudioside D. In one embodiment, the steviol glycoside blend contains 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% rebaudioside D.

[0101] In yet another embodiment, the sweetener is a steviol glycoside blend containing a sweetening amount of rebaudioside N. In one embodiment, the steviol glycoside blend contains 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% rebaudioside N.

[0102] In another embodiment, the sweetener is a steviol glycoside blend containing a sweetening amount of rebaudioside O. In one embodiment, the steviol glycoside blend contains 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% rebaudioside O.

[0103] In another embodiment, the sweetener is a steviol glycoside blend containing a sweetening amount of rebaudioside J. In one embodiment, the steviol glycoside blend contains 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% rebaudioside J.

[0104] Stevioside blend sweeteners typically have a total steviol glycoside content of about 95% or more on a dry basis by weight. The remaining 5% comprises other non-stevioside compounds, such as byproducts from extraction or purification processes. In some embodiments, the steviol glycoside blend sweetener has a total steviol glycoside content of about 96% or more, about 97% or more, about 98% or more, or about 99% or more. As used herein, "total steviol glycoside content" refers to the sum of the relative weight contributions of each steviol glycoside in the sample.

[0105] In one embodiment, the sweetener comprises at least one mogroside, wherein the at least one mogroside is present in a sweetening amount. Mogroside can be natural, synthetic, or a combination of natural and synthetic.

[0106] Mogrosides can be provided in pure form or as part of a mixture (i.e., mogroside blends). Exemplary mogrosides include, but are not limited to, any of the following: grosmogroside I, mogroside IA, mogroside IE, 11-oxomogroside IA, mogroside II, mogroside II A, mogroside II B, and mogroside II. E, 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-deoxymogroside V, 7-oxomogroside V, 11-oxomogroside V, isomogroside V, mogroside VI, mogroside alcohol, 11-oxomogroside alcohol, symbioside I, isomers of symbioside I (e.g., those disclosed in 20170119032; the patent is incorporated herein by reference in its entirety), and especially the 1,6-α isomers of symbioside I (Examples 7 and 10) and combinations thereof.

[0107] Additional exemplary mogrosides include those described in U.S. Patent Application Publication 2016039864, the contents of which are incorporated herein by reference. In specific embodiments, mogrosides are selected from (3β,9β,10α,11α,24R)-3-[(4-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosyl]-25-hydroxy-9-methyl-19-norlanost-5-en-24-yl-[2-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosyl]-β-D-glucopyranoside); (3β,9β,10α,11α,24R)-[(2-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]-25-hydroxy (3β,9β,10α,11α,24R)-[(2-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosyl]-β-D-glucopyranoside); (3β,9β,10α,11α,24R)-[(2-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]-25-hydroxy-9-methyl-19-norlanost-5-en-24-yl-[2-O-β-D-glucopyranosyl-6-O-β-D-glucopyranosyl]-β-D-glucopyranoside and combinations thereof.

[0108] The mogroside blend contains at least one mogroside for sweetening.

[0109] In some embodiments, the mogroside blend contains 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% mogroside.

[0110] In other embodiments, the mogroside blends have a total mogroside content of about 95% or more on a dry basis by weight. In some embodiments, the mogroside blends have 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.

[0111] In one embodiment, the sweetener is a mogroside blend containing a sweetening amount of sarcoside I. In one embodiment, the mogroside blend contains 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% sarcoside I.

[0112] In another embodiment, the sweetener is a mogroside blend, the blend comprising a sweetening amount of the 1,6-α isomer of symbioside I. In one embodiment, the mogroside blend 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 1,6-α isomer of symbioside I.

[0113] In yet another embodiment, the sweetener is a mogroside blend containing a sweetening amount of mogroside V. In one embodiment, the mogroside blend contains 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% mogroside V.

[0114] The weight ratio of the at least one sweetener to the at least one dihydrochalcone compound described herein can vary. Typically, the weight ratio of the at least one sweetener to the at least one dihydrochalcone 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 ... 0:1 to about 100:1, from about 500:1 to about 50:1, from about 500:1 to about 25:1, from about 500:1 to about 10:1, from about 400:1 to about 300:1, from about 400:1 to about 200:1, from about 400:1 to about 100:1, from about 400:1 to about 50:1, from about 400:1 to about 25:1, from about 400:1 to about 10:1, from about 400:1 to about 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.

[0115] IV. Composition

[0116] The present invention also provides compositions comprising at least one sweetener described herein and at least one dihydrochalcone compound having formula I.

[0117] The at least one dihydrochalcone compound described herein is present in the composition such that when the composition is added to a consumer product, the sucrose equivalent of the consumer product is increased by an amount present in the composition compared to a consumer product in the absence of the dihydrochalcone compound.

[0118] The at least one dihydrochalcone described herein may be present in the composition in such a quantity that when the composition is added to a consumer product, the sucrose equivalent (SE) of the consumer product is increased by at least about 1.2 times, such as at least about 1.5 times, at least about 1.7 times, at least about 2.0 times, at least about 2.5 times, at least about 3.0 times, or at least about 4.0 times.

[0119] The amount of sucrose in a reference solution can be described using Brix (°Bx), and thus another measure of sweetness. One Brix is ​​equal to 1 gram of sucrose in 100 grams of solution, and is expressed as a weight percentage (%w / w) of the strength of the solution (strictly speaking, by mass).

[0120] In some embodiments, the at least one dihydrochalcone compound described herein is present in the composition in an amount such that when the composition is added to a consumer product, the consumer product has a sweetness equivalent to at least about 8 degrees Brix, such as about 8 degrees Brix, about 9 degrees Brix, about 10 degrees Brix, about 11 degrees Brix, or about 12 degrees Brix.

[0121] In other embodiments, the at least one dihydrochalcone compound described herein is present in the composition in an amount such that when the composition is added to a consumer product, the Brix level of the consumer product increases by at least 1 degree Brix, such as at least 2 degrees Brix, at least 3 degrees Brix, at least 4 degrees Brix, or at least 5 degrees Brix. In other embodiments, the at least one dihydrochalcone compound described herein is present in the composition such that when the composition is added to a consumer product, it modifies one or more flavor properties of the sweetener, thereby making the consumer product taste more like a sucrose-sweetened product compared to the same one or more flavor properties of a consumer product in the absence of the at least one dihydrochalcone. Exemplary flavor property modulation includes 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 sweeteners can be simultaneously modulated, resulting in consumer products with a greater overall sucrose sweetening characteristic. Methods for quantifying the improvement of sucrose sweetening characteristics are known in the art and include, for example, taste testing and histogram plotting.

[0122] It should be noted that the comparison is made between (i) a consumer product containing at least one of the sweeteners and at least one of the dihydrochalcone compounds described herein and (ii) a consumer product containing at least one sweetener but not the at least one of the dihydrochalcone compounds described herein.

[0123] Consumer products containing at least one sweetener and at least one dihydrochalcone compound described herein are also provided.

[0124] The at least one dihydrochalcone compound described herein is typically present in a consumer product in an amount that effectively enhances the sweetness of the product and / or modulates one or more flavor properties of the sweetener to make the product taste more like sucrose. In one embodiment, the at least one dihydrochalcone compound described herein is present at or below the sweetness recognition threshold concentration of dihydrochalcone, i.e., an amount that does not provide sweetness when tasted alone.

[0125] Typically, the at least one dihydrochalcone compound described herein is present in the consumer product in an amount that effectively enhances the sucrose equivalent (SE) of the consumer product by at least about 1.2 times, 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.

[0126] In some embodiments, the at least one dihydrochalcone compound described herein is present in an amount that effectively provides a sweetness equivalent to at least about 8 degrees Brix, such as about 8 degrees Brix, about 9 degrees Brix, about 10 degrees Brix, about 11 degrees Brix, or about 12 degrees Brix.

[0127] In other embodiments, the at least one dihydrochalcone compound described herein is present in an amount that effectively increases the Brix level of the consumer product by at least 1 degree compared to the Brix level of the consumer product in the absence of the at least one dihydrochalcone, such as at least 2 degrees Brix, at least 3 degrees Brix, at least 4 degrees Brix, or at least 5 degrees Brix.

[0128] In other embodiments, the amount of the at least one dihydrochalcone compound of Formula I present in the consumer product is such that it effectively modulates one or more flavor properties of the sweetener to make the consumer product taste more like sucrose-sweetened sweetness. Exemplary flavor property modulation includes 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, thus making the consumer product generally more sucrose-sweetened. Methods for quantifying the improvement of sucrose-sweetened characteristics are known in the art and include taste testing and histogram plotting.

[0129] The specific concentrations of the at least one dihydrochalcone compound and sweetener described herein will vary depending on the specific dihydrochalcone compound and sweetener.

[0130] In one embodiment, the at least one dihydrochalcone compound described herein is present in the consumer product at a concentration from about 1 ppm to about 50 ppm, such as from about 1 ppm to about 45 ppm, from about 1 ppm to about 40 ppm, from about 1 ppm to about 35 ppm, from about 1 ppm to about 30 ppm, from about 1 ppm to about 25 ppm, from about 1 ppm to about 20 ppm, from about 1 ppm to about 15 ppm, from about 1 ppm to about 10 ppm, and from about 1 ppm to about 5 ppm. In another embodiment, the at least one dihydrochalcone is present in the consumer product at a concentration from about 5 ppm to about 40 ppm, such as from about 5 ppm to about 35 ppm, from about 5 ppm to about 20 ppm, from about 20 ppm to about 40 ppm, from about 20 ppm to about 30 ppm, or from about 30 ppm to about 40 ppm.

[0131] In one embodiment, the at least one sweetener described herein is in the range of about 50 ppm to about 600 ppm, such as 50 ppm to about 500 ppm, about 50 ppm to about 400 ppm, about 50 ppm to about 300 ppm, about 50 ppm to about 300 ppm, about 50 ppm to about 200 ppm, about 50 ppm to about 100 ppm, about 100 ppm to about 600 ppm, about 100 ppm to about 500 ppm, about 100 ppm to about 400 ppm, about 100 ppm to about The concentrations of 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, and about 500 ppm to about 600 ppm are present in consumer products.

[0132] The weight ratio of at least one sweetener to at least one dihydrochalcone described herein may also vary, as discussed above.

[0133] In one specific embodiment, a consumer product is provided comprising at least one dihydrochalcone compound described herein and at least one sweetener, wherein the at least one dihydrochalcone is present in an amount that effectively enhances the SE of the consumer product by at least about 1.2 times compared to the SE of a consumer product in the absence of the at least one dihydrochalcone.

[0134] In one embodiment, the consumer product with enhanced sweetness has about 2% (w / v) or more, such as about 3% or more, about 4% or more, about 5% or more, about 6% or more, about 7% or more, about 8% or more, about 9% or more, about 10% or more, about 11% or more, about 12% or more, about 13% or more, or about 14% or more SE.

[0135] In another embodiment, the consumer product with enhanced sweetness has a Brix level of about 3 to about 12, such as about 3 Brix or greater, about 4 Brix or greater, about 5 Brix or greater, about 5 Brix or greater, about 7 Brix or greater, about 8 Brix or greater, about 9 Brix or greater, about 10 Brix or greater, and about 11 Brix or greater.

[0136] Exemplary consumer products include, but are not limited to, 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.

[0137] Consumer products may optionally include one or more additional additives. Exemplary additives include, but are not limited to, carbohydrates, polyols, amino acids and their corresponding salts, polyamino acids and their corresponding salts, glycosaminoglycans and their corresponding salts, nucleotides, organic acids, inorganic acids, organic salts (including organic acid salts and organic base salts), inorganic salts, bitter compounds, flavoring agents and flavoring components, astringent compounds, proteins or protein hydrolysates, surfactants, emulsifiers, weighing agents, gums, antioxidants, pigments, flavonoids, alcohols, polymers, and combinations thereof.

[0138] In one embodiment, the consumer product 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.

[0139] 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 carbohydrates that can be reduced without adversely affecting the taste.

[0140] 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 salt forms such as sodium or potassium salts or acid salts. Amino acid additives may also be in D- or L-configuration and in mono-, di-, or ternary forms of the same or different amino acids. Additionally, if appropriate, these amino acids may 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 in D- or L-configuration. 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 such that at least one atom is added, removed, substituted, or a combination thereof (e.g., N-alkyl polyamino acids or N-acyl polyamino acids). As used herein, polyamino acids encompass both modified and unmodified polyamino acids.For example, modified polyamino acids include, but are not limited to, polyamino acids 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.

[0141] 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.

[0142] 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 include nucleotide-related additives, such as nucleosides or nucleic acid bases (e.g., guanine, cytosine, adenine, thymine, uracil).

[0143] 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.

[0144] 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, carboxamido, phosphonyl, oxyphosphonyl, phosphoryl, phosphonyl, thioester, thioether, acid anhydride, oxime, hydrazine, carbamoyl, phosphorus, or phosphonate. In specific embodiments, when present in consumer products (e.g., beverages), the organic acid additive is present in the sweetener composition in an amount that effectively provides a concentration from about 10 ppm to about 5,000 ppm.

[0145] 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 or alkaline earth metal salts (e.g., inositol hexaphosphate Mg / Ca).

[0146] Suitable bitter compound additives include, but are not limited to, caffeine, quinine, urea, bitter orange oil, naringin, quassula and its salts.

[0147] 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 TMBitterness inhibitors 1, 2, 9 and 10 (Natural Advantage) TM Freehold, New Jersey, USA and Sucramask TM (Creative Research Management, Stockton, California, USA)

[0148] Suitable polymer additives include, but are not limited to, chitosan, pectin, pectinic acid, polyuronic acid, polygalacturonic acid, starch, food hydrolysates or their crude extracts (e.g., Senegalese gum arabic (Fibergum arabic)). TM (e.g., seleno-arabinose, 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.

[0149] 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).

[0150] 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, hexadecylpyridine 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.

[0151] 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.

[0152] Suitable alcohol additives include, but are not limited to, ethanol. In specific embodiments, the alcohol additive is present in the consumer product at a concentration from about 625 ppm to about 10,000 ppm.

[0153] 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). The astringent additives are present in the sweetening composition at a concentration from about 10 ppm to about 5,000 ppm.

[0154] The consumer products 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, weight management agents, osteoporosis management agents, phytoestrogens, long-chain primary aliphatic saturated alcohols, phytosterols, and combinations thereof.

[0155] 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 primary aliphatic saturated alcohols, phytosterols, and combinations thereof.

[0156] Examples of suitable antioxidants used in embodiments of the present invention include, but are not limited to, vitamins, vitamin cofactors, minerals, hormones, carotenoids, carotenoid terpenes, non-carotenoid terpenes, flavonoids, flavonoid polyphenols (such as bioflavonoids), flavonols, flavonoids, phenols, polyphenols, phenolic esters, polyphenolic esters, non-flavonoid phenols, isothiocyanates, and combinations thereof. In some embodiments, antioxidants are vitamin A, vitamin C, vitamin E, ubiquinone, minerals selenium, 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. gutamine), oxalic acid, tocopherol derivatives, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediaminetetraacetic acid (EDTA), tert-butylhydroquinone, acetic acid, pectin, tocotrienol, tocopherol, coenzyme Q10, zeaxanthin, astaxanthin, canthaxanthin, saponins, limonene, kaempferol, myricetin, isorhamnetin, proanthocyanidins, quercetin, rutin, luteolin, apigenin, tangeritin, hesperidin, grapefruit Cortisol, 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, anythocyanins, cyaniding, delphinidin, malvidin, malvidin, methyl anthocyanins, petunidin, ellagic acid, and other similar compounds. Gallic acid, salicylic acid, rosmarinic acid, cinnamic acid and its derivatives (e.g., ferulic acid), chlorogenic acid, chicoric acid, gall tannins, ellagic acid, flavonoids, β-anthocyanins 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 suitable sources of 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.

[0157] Specific antioxidants belong to the class of phytonutrients called polyphenols (also known as "polyphenols"), a group of chemical substances found in plants, characterized by the presence of more than one phenolic group per molecule. 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.

[0158] In specific embodiments, the antioxidant is a catechin, such as epigallocatechin gallate (EGCG). Suitable sources of catechins used in embodiments of the present invention include, but are not limited to, green tea, white tea, black tea, oolong tea, chocolate, cocoa, red wine, grape seeds, red grape skins, purple grape skins, red grape juice, purple grape juice, berries, pycnogenol, and red apple skins.

[0159] In some embodiments, the antioxidant is selected from proanthocyanidins, proanthocyanidins, or combinations thereof. Suitable sources of proanthocyanidins and proanthocyanidins used in embodiments of the present invention include, but are not limited to, red grapes, purple grapes, cocoa, chocolate, grape seeds, red wine, cocoa beans, cranberries, apple peels, plums, blueberries, blackcurrants, chokeberries, green tea, sorghum, cinnamon, barley, red kidney beans, black and white bean beans, hops, almonds, hazelnuts, pecans, pistachios, pycnogenol, and amaranth.

[0160] In a specific embodiment, the antioxidant is anthocyanin. Suitable sources of anthocyanins used in embodiments of the present invention include, but are not limited to, raspberries, blueberries, blueberries, cranberries, raspberries, cherries, pomegranates, strawberries, elderberries, rowan fruits, red grape skins, purple grape skins, grape seeds, red wine, blackcurrants, red currants, cocoa, plums, apple peels, peaches, red pears, red cabbage, red onions, red oranges, and blackberries.

[0161] In some embodiments, the antioxidant is selected from quercetin, rutin, or a combination thereof. Suitable sources of quercetin and rutin used in embodiments of the present invention include, but are not limited to, red apples, onions, kale, blueberries, bilberries, rowan fruit, cranberries, blackberries, blueberries, strawberries, raspberries, blackcurrants, green tea, black tea, plums, apricots, parsley, leeks, broccoli, red peppers, berry wines, and ginkgo.

[0162] In some embodiments, the antioxidant is resveratrol. Suitable sources of resveratrol used in embodiments of the present invention include, but are not limited to, red grapes, peanuts, cranberries, blueberries, blueberries, mulberries, Japanese Itadoritea, and red wine.

[0163] In a specific embodiment, the antioxidant is isoflavone. 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.

[0164] In some embodiments, the antioxidant is curcumin. Suitable sources of curcumin used in embodiments of the invention include, but are not limited to, turmeric and mustard.

[0165] In specific embodiments, the antioxidant is selected from quercetin, ellagitannins, or combinations thereof. Suitable sources of quercetin and ellagitannins used in embodiments of the present invention include, but are not limited to, pomegranate, raspberry, strawberry, walnut, and aged red wine.

[0166] In some embodiments, the antioxidant is a citrus flavonoid, such as hesperidin or naringin. Suitable sources of citrus flavonoids such as hesperidin or naringin used in embodiments of the invention include, but are not limited to, oranges, grapefruits, and citrus juices.

[0167] In a specific embodiment, the antioxidant is chlorogenic acid. Suitable sources of chlorogenic acid used in embodiments of the present invention include, but are not limited to, raw coffee, Paraguayan tea, red wine, grape seeds, red grape skins, purple grape skins, red grape juice, purple grape juice, apple juice, cranberries, pomegranates, blueberries, strawberries, sunflowers, echinacea, pycnogenol, and apple peels.

[0168] Suitable dietary fibers include, but are not limited to, non-starch polysaccharides, lignin, cellulose, methylcellulose, hemicellulose, β-glucan, pectin, gums, mucilage, waxes, inulin, oligosaccharides, fructooligosaccharides, cyclodextrin, chitin, and combinations thereof.

[0169] Food sources of dietary fiber include, but are not limited to, grains, legumes, fruits, and vegetables. Grains that provide dietary fiber include, but are not limited to, oats, rye, barley, and wheat. Legumes that provide fiber include, but are not limited to, peas and beans such as soybeans. Fruits and vegetables that provide fiber include, but are not limited to, apples, oranges, pears, bananas, berries, tomatoes, green beans, broccoli, cauliflower, carrots, potatoes, and celery. Plant-based foods such as bran, nuts, and seeds (such as flaxseed) are also sources of dietary fiber. Plant parts that provide dietary fiber include, but are not limited to, stems, roots, leaves, seeds, pulp, and peel.

[0170] Fatty acids are any straight-chain monocarboxylic acids and include 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. Suitable ω-3 fatty acids include, but are not limited to, linolenic acid, α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, octadecanoic acid, eicosapentaenoic acid, and combinations thereof. Suitable ω-6 fatty acids include, but are not limited to, linoleic acid, gamma-linolenic acid, dihomo-gamma-linolenic acid, arachidonic acid, eicosadienoic acid, docosadienoic acid, adrenaline, docosapentaenoic acid, and combinations thereof. Suitable esterified fatty acids used in embodiments of the invention 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.

[0171] Suitable vitamins include vitamins A, D, E, K, B1, B2, B3, B5, B6, B7, B9, B12, and C. Many other compounds have been officially classified as vitamins. These compounds can be called pseudovitamins and include, but are not limited to, compounds such as ubiquinone (coenzyme Q10), panthenine, dimethylglycine, taestrile, amygdalin, flavonoids, p-aminobenzoic acid, adenine, adenosine, and S-methylmethionine. As used herein, the term vitamin includes pseudovitamins.

[0172] Minerals are 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.

[0173] In other specific embodiments of the invention, the minerals are trace minerals considered essential for human nutrition, and non-limiting examples include bismuth, boron, lithium, nickel, rubidium, silicon, strontium, tellurium, tin, titanium, tungsten, and vanadium.

[0174] 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. Sulfites include, but are not limited to, sulfur dioxide, sodium bisulfite, and potassium bisulfite. Propionates include, but are not limited to, propionic acid, calcium propionate, and sodium propionate. Benzoates include, but are not limited to, sodium benzoate and benzoic acid. Sorbicates include, but are not limited to, potassium sorbate, sodium sorbate, calcium sorbate, and sorbic acid. Nitrates and nitrites include, but are not limited to, sodium nitrate and sodium nitrite. In another specific embodiment, the at least one preservative is a bacteriocin, such as nisin. In another specific embodiment, the preservative is ethanol. In yet another specific embodiment, the preservative is ozone. Anti-enzyme agents suitable for use as preservatives in specific embodiments of the present invention include ascorbic acid, citric acid, and metal chelating agents such as ethylenediaminetetraacetic acid (EDTA).

[0175] The hydration product may be an electrolyte, and non-limiting examples include sodium, potassium, calcium, magnesium, chloride, phosphate, bicarbonate, 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. 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 specific embodiments of the invention, the hydration product is a carbohydrate that replenishes the energy stores burned by muscles. Suitable carbohydrates for 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 used in specific embodiments include trioses, tetroses, pentoses, hexoses, heptoses, octoses, and nonoses. Non-limiting examples of specific types of suitable monosaccharides include glyceraldehyde, dihydroxyacetone, erythrose, thoraxose, erythulose, arabinose, lysose, ribose, xylose, ribulose, xylulose, allose, azoose, galactose, glucose, gulose, idulose, mannose, talose, fructose, allulose, sorbitolose, tagatose, mannohepulose, 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, fruit juice, or tea. In another specific embodiment, hydration is provided by flavanols that offer cell rehydration. 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. In a specific embodiment, the hydration product is a glycerol solution that enhances athletic endurance.

[0176] Probiotics include microorganisms that are beneficial to health when consumed in effective amounts. Probiotics can include, but are not limited to, bacteria, yeast, and fungi. Examples of probiotics include, but are not limited to, bacteria of the genus *Lactobacilli*, *Bifidobacteria*, *Streptococci*, or combinations thereof. In a specific embodiment of the invention, the at least one probiotic is selected from the genus *Lactobacilli*. *Lactobacilli* (i.e., bacteria of the genus *Lactobacilli*, hereinafter "L.") Non-limiting examples of lactobacilli species found in the human gastrointestinal tract include *Lactobacillus acidophilus*, *Lactobacillus casei*, *Lactobacillus fermentum*, *Lactobacillus saliva*, *Lactobacillus brevis*, *Lactobacillus leichmannii*, *Lactobacillus plantarum*, *Lactobacillus cellobiosus*, *Lactobacillus reuteri*, *Lactobacillus rhamnosus*, *Lactobacillus GG*, *Lactobacillus bulgaricus*, and *Lactobacillus thermophilus*. According to other specific embodiments of the invention, the probiotics are selected from the genus *Bifidobacterium*.Non-restricted species of Bifidobacteria found in the human gastrointestinal tract include *Bifidobacterium anguulatum*, *Bifidobacterium animalis*, *Bifidobacterium asteroides*, *Bifidobacterium bifidum*, *Bifidobacterium boum*, *Bifidobacterium breve*, *Bifidobacterium catenulatum*, *Bifidobacterium choerinum*, *Bifidobacterium coryneforme*, *Bifidobacterium cuniculi*, *Bifidobacterium dentium*, *Bifidobacterium gallicum*, *Bifidobacterium gallinarum*, and *Bifidobacterium chrysanthemum*. Bifidobacterium indicum, Bifidobacterium longum, Bifidobacterium magnum, Bifidobacterium merycicum, Bifidobacterium minimum, Bifidobacterium pseudodocatenulatum, Bifidobacterium pseudodolongum, Bifidobacterium psychraerophilum, Bifidobacterium pullorum, Bifidobacterium ruminantium, Bifidobacterium saeculare, Bifidobacterium scarovii, Bifidobacterium simiae, Bifidobacterium subtile, Bifidobacterium thermophilum, Bifidobacterium urinalis, and some other Bifidobacterium species. According to other specific embodiments of the invention, the probiotics are selected from the genus Streptococcus. Streptococcus thermophilus is a Gram-positive facultative anaerobic bacterium. Other non-restricted probiotic species of this bacterium include Streptococcus salivarus and Streptococcus cremoris.

[0177] Prebiotics are compositions that promote the growth of beneficial bacteria in the gut. Prebiotics include, but are not limited to, mucopolysaccharides, oligosaccharides, polysaccharides, amino acids, vitamins, nutrient precursors, proteins, and combinations thereof. According to specific embodiments of the 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 the invention include fructooligosaccharides, inulin, isomaltooligosaccharides, lactitol, lactulose oligosaccharides, lactulose, dextrin, soybean oligosaccharides, trans-galactosidase, and xylooligosaccharides. According to other specific embodiments of the invention, prebiotics are amino acids.

[0178] As used herein, “weight management agents” include appetite suppressants and / or thermic agents. As used herein, the phrases “appetite suppressant,” “appetite-satiety composition,” “satiety agent,” and “satiety ingredient” are synonymous. The phrase “appetite suppressant” describes macronutrients, herbal extracts, exogenous hormones, appetite suppressants, anorexia nervosa, drugs, and combinations thereof that, when delivered in effective amounts, inhibit, prohibit, reduce, or otherwise diminish a person’s appetite. The phrase “thermic agent” describes macronutrients, herbal extracts, exogenous hormones, appetite suppressants, anorexia nervosa, drugs, and combinations thereof that, when delivered in effective amounts, stimulate or otherwise enhance a person’s thermic action or metabolism.

[0179] Suitable weight management agents include macronutrients selected from the following groups: protein, carbohydrates, dietary fat, and combinations thereof. Carbohydrates generally include sugars, starches, cellulose, and gums that are converted into glucose by the body for energy. Non-limiting examples of 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. Dietary fat is a lipid consisting of a combination of saturated and unsaturated fatty acids. Polyunsaturated fatty acids have been shown to have a greater satiating effect than monounsaturated fatty acids. Therefore, the dietary fats presented herein ideally include polyunsaturated fatty acids, non-limiting examples of which include triacylglycerols.

[0180] In one specific embodiment, the weight management agent is an herbal extract. 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. In one specific embodiment, the herbal extract is derived from plants of the genus *Hypericum*, which includes 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*. Plants of the genus *H.* are succulent plants native to South Africa. 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*, namely the family Asclepiadaceae. In another specific embodiment, the at least one herbal extract is derived from plants of the genus *Caralluma*.The genus *T. piliferum* consists of succulent plants typically native to South Africa, similar to the genus *T. pyridae*, and includes *T. officinale*. In another specific embodiment, the herbal extract is derived from the genera *S. gigantean* or *Obedia*, whose species include *S. gigantean* and *O. variegate*, respectively. Both *S. gigantean* and *Obedia* belong to the subtribe *Asclepiadaceae*, the same family as *T. pyridae*. In another specific embodiment, the herbal extract is derived from the genus *Aleurone*. *Aleurone* also belongs to the tribe *Asclepiadaceae*. Non-limiting examples of *Aleurone* include *A. incarnate*, *A. curassayica*, *A. syriaca*, and *A. tuberose*. Without being bound by any theory, these extracts are believed to contain steroidal compounds with appetite-suppressing effects, such as pregnaneglycosides and pregnanegenins. 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.

[0181] In some embodiments, the osteoporosis treatment agent is at least one calcium source, i.e., 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. According to a specific embodiment, the osteoporosis management agent is a magnesium source, i.e., 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 includes amino acid chelated magnesium or creatine chelated magnesium. In other embodiments, the osteoporosis agent is selected from vitamins D, C, K, their precursors, and / or β-carotene, and combinations thereof. Many plants and plant extracts have also been identified as effective in preventing and treating osteoporosis. Without wishing to be bound by any theory, it is believed that these plants and plant extracts stimulate osteoblasts and / or inhibit bone resorption, thereby promoting bone regeneration and strength. 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*, *Juniperus*, and *Prunus* as disclosed in U.S. Patent Publication No. 2005 / 0079232. Species from the genera *Prunus*, *Iris*, *Cichorium*, *Dodonaea*, *Epimedium*, *Erigonoum*, *Soya*, *Mentha*, *Ocimum*, *thymus*, *Tanacetum*, *Plantago*, *Spearmint*, *Bixa*, *Vitis*, *Rosemarinus*, *Rhus*, and *Anethum*.

[0182] Examples of suitable phytoestrogens used in embodiments of the present invention include, but are not limited to, isoflavones, stilbenes, lignans, resorcyclic acid lactone, coumarin, coumestan, coumestrol, equol, and combinations thereof. 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 in each molecule. Suitable phytoestrogens isoflavones according to embodiments of the present invention include genistein, daidzein, genistein, chickpea glycoside A, gentiocarbazone, their respective naturally occurring glycosides and glycoside conjugates, maltocarbazone, open-ring isolar resinin, enteroester, enterodiol, plant tissue proteins, and combinations thereof.

[0183] Long-chain aliphatic saturated primary alcohols are a group of organic compounds. The term long-chain refers to the fact that these compounds contain at least eight carbon atoms. Non-limiting examples of specific long-chain aliphatic saturated primary alcohols used in specific embodiments of the invention include 8-carbon 1-octanol, 9-carbon 1-nonanol, 10-carbon 1-decanol, 12-carbon 1-dodecanol, 14-carbon 1-tetradecanol, 16-carbon 1-hexadecanol, 18-carbon 1-octadecanol, 20-carbon 1-eicosanool, 22-carbon 1-docodecanol, 24-carbon 1-tetracosanool, 26-carbon 1-hexadecanol, 27-carbon 1-heptadecanol, 28-carbon 1-octanosol, 29-carbon 1-nonadecanol, 30-carbon 1-triacontanol, 32-carbon 1-triadecanol, and 34-carbon 1-triadecanol. In a particularly desirable embodiment of the invention, the long-chain primary aliphatic saturated alcohol is prilool. Priligol is a term referring to a mixture of long-chain aliphatic saturated primary alcohols, mainly composed of: 28-carbon 1-octacosanol and 30-carbon 1-triacontanol, as well as other alcohols in lower concentrations such as 22-carbon 1-docosahexacosanol, 24-carbon 1-tetracosanol, 26-carbon 1-hexacosanol, 27-carbon 1-heptacosanol, 29-carbon 1-nonacosanol, 32-carbon 1-trimosahexacosanol, and 34-carbon 1-trimosahexacosanol.

[0184] 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).

[0185] According to specific embodiments of the present invention, non-limiting examples of phytosterols include β-sitosterol, campesterol, cycloartenol, and saturated forms of other triterpenoid alcohols.

[0186] 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). The phytosterols and phytosterols of the present invention may also be in their ester forms. 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.

[0187] 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.

[0188] In one embodiment, the consumer product is a beverage or beverage product.

[0189] As used herein, “beverage product” refers to ready-to-drink beverages, beverage concentrates, beverage syrups, or beverage powders. 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 root juice ale. Non-carbonated beverages include, but are not limited to, fruit juices, fruit-flavored 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 (e.g., milk drinks, milk-containing coffee, café au lait, milk tea, fruit milk drinks), beverages containing cereal extracts, and smoothies.

[0190] 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. Solid 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.

[0191] The beverage contains a matrix, which is the basic component in which these ingredients (including the compositions of the present invention) are dissolved. In one embodiment, the beverage contains beverage-quality water as a matrix, such as deionized water, distilled water, reverse osmosis water, carbon-treated water, pure water, softened water, and combinations thereof. Additional suitable matrices include, but are not limited to, phosphoric acid, phosphate buffer, citric acid, citrate buffer, and carbon-treated water.

[0192] The beverage or beverage product may further comprise at least one additional sweetener. Any sweetener detailed herein may be used, including natural, non-natural, or synthetic sweeteners. In some embodiments, the beverage or beverage product does not contain a carbonated sweetener.

[0193] Considering that the pH of a consumer product (e.g., a beverage) does not substantially or adversely affect the taste of a sweetener, a non-limiting example of a beverage's pH range could be from about 1.8 to about 10. Another example includes a pH range from about 2 to about 5. In a specific embodiment, the pH of the beverage could be from about 2.5 to about 4.2. Those skilled in the art will understand that the pH of a beverage can vary depending on the type of beverage. For example, dairy beverages can have a pH greater than 4.2.

[0194] The titratable acidity of a beverage can range, for example, from about 0.01% to about 1.0% by weight of the beverage.

[0195] 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.

[0196] 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).

[0197] The beverage may be caffeinated or decaffeinated.

[0198] 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.

[0199] The beverage can be a high-calorie drink, with up to about 120 calories per 8-ounce serving.

[0200] The beverage can be a medium-calorie drink, with up to about 60 calories per 8-ounce serving.

[0201] The beverage can be a low-calorie drink, with up to about 40 calories per 8-ounce serving.

[0202] The beverage can be a zero-calorie drink, which has less than about 5 calories per 8-ounce serving.

[0203] In one specific embodiment, the consumer product is a cola beverage. The cola beverage may be a low-calorie, medium-calorie, or zero-calorie beverage.

[0204] 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.

[0205] In one specific embodiment, the beverage of the present invention is a flavored water beverage.

[0206] V. Usage Instructions

[0207] A method is provided to enhance the sweetness of a consumer product and / or modulate one or more flavor properties of a sweetener to make the consumer product taste more like a sucrose-sweetened consumer product.

[0208] 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 dihydrochalcone compound having formula I to the consumer product to provide a consumer product with enhanced sweetness.

[0209] 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 dihydrochalcone compound having Formula I to the consumer product matrix to provide a consumer product with enhanced sweetness. The at least one sweetener and the at least one dihydrochalcone compound having Formula I may be added together, i.e., added as a composition, or added separately.

[0210] As used herein, the term “consumer product matrix” refers to a consumer product containing all the typical ingredients except for sweeteners or sweetener components.

[0211] In one specific embodiment, the SE of a consumer product comprising at least one dihydrochalcone compound of formula I 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 the at least one dihydrochalcone compound of formula I, 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.

[0212] In another embodiment, adding at least one dihydrochalcone compound of formula I to a consumer product or a consumer product matrix increases the Brix level by at least 1 degree, such as at least 2 degrees, at least 3 degrees, or at least 4 degrees, compared to the Brix level of a consumer product that does not contain a dihydrochalcone compound of formula I.

[0213] In one specific embodiment, the consumer product is a beverage.

[0214] Therefore, methods for enhancing the sweetness of beverages include (i) providing a beverage containing at least one sweetener and (ii) adding at least one dihydrochalcone compound having formula I to a beverage to provide a beverage with enhanced sweetness.

[0215] In another embodiment, a method for enhancing the sweetness of a beverage includes (i) providing a beverage base and (ii) adding at least one sweetener and at least one dihydrochalcone compound having Formula I to the beverage base to provide a beverage with enhanced sweetness. The at least one sweetener and the at least one dihydrochalcone compound having Formula I may be added together, i.e., added as a composition, or added separately.

[0216] In another aspect, a method for making a consumer product taste more like sucrose-sweetened includes (i) providing a consumer product containing at least one sweetener and (ii) adding one or more flavor properties of the sweetener to make the consumer product taste more like sucrose-sweetened compared to a consumer product in the absence of at least one dihydrochalcone compound of formula I.

[0217] In another embodiment, a method for making a consumer product taste more like sucrose-sweetened includes (i) providing a consumer product matrix and (ii) adding at least one sweetener and at least one dihydrochalcone compound of formula I to the consumer product matrix to provide a consumer product that tastes more like sucrose-sweetened, wherein the at least one dihydrochalcone compound of formula I is present to effectively modulate one or more flavor properties of the sweetener to a level that makes the consumer product taste more like sucrose-sweetened compared to a consumer product in the absence of the at least one dihydrochalcone compound of formula I. The at least one sweetener and the at least one dihydrochalcone compound of formula I may be added together, i.e., added as a composition, or added separately.

[0218] As discussed above, in the method of the present invention, at least one dihydrochalcone compound having Formula I may be present in the consumer product at the identified concentration.

[0219] Methods for preparing consumer products with enhanced sweetness are also provided.

[0220] In one aspect, a method of preparing a consumer product includes (i) providing a consumer product containing at least one sweetener and (ii) adding at least one dihydrochalcone compound having formula I to the consumer product to provide the consumer product.

[0221] In another aspect, a method for preparing a consumer product is provided, the method comprising (i) providing a consumer product matrix and (ii) adding at least one sweetener and at least one dihydrochalcone compound having formula I to the consumer product matrix to provide the consumer product. The at least one sweetener and the at least one dihydrochalcone compound having formula I may be added together, i.e., added as a composition, or added separately.

[0222] In another aspect, a method for preparing a consumer product is provided, the method comprising (i) providing a consumer product matrix and (ii) adding a composition comprising at least one sweetener and at least one dihydrochalcone compound having formula I to the consumer product matrix to provide the consumer product.

[0223] In another aspect, a method for preparing a beverage is provided, the method comprising (i) providing a beverage matrix and (ii) adding at least one sweetener and at least one dihydrochalcone compound having formula I to the beverage matrix to provide the beverage. The at least one sweetener and the at least one dihydrochalcone compound having formula I may be added together, i.e., added as a composition, or added separately.

[0224] In another aspect, a method for preparing a beverage is provided, the method comprising (i) providing a beverage matrix and (ii) adding a composition comprising at least one sweetener and at least one dihydrochalcone compound having formula I to the beverage matrix to provide the beverage.

[0225] In another aspect, a method for preparing a sweetened beverage is provided, the method comprising (i) providing an unsweetened beverage and (ii) adding at least one sweetener and at least one dihydrochalcone compound having formula I to the unsweetened beverage to provide a sweetened beverage. The at least one sweetener and the at least one dihydrochalcone compound having formula I may be added together, i.e., added as a composition, or added separately.

[0226] In another aspect, a method for preparing a sweetened beverage is provided, the method comprising (i) providing an unsweetened beverage and (ii) adding a composition comprising at least one sweetener and at least one dihydrochalcone compound having formula I to the unsweetened beverage to provide a sweetened beverage.

[0227] Example 1: Isolation and characterization of analogues from *Stachys rubrum*

[0228] instrument: Record 1D and 2D NMR data on a Bruker Avance III 500 or 600HD spectrometer, and 1 H and 13 C NMR chemical shift reference methanol-d4 (δH 3.31 and δ C 49.0) and DMSO-d6 (δH 2.50 and δ CThe residual solvent peak was observed at 39.52. High-resolution electrospray ionization mass spectrometry (HRESIMS) was performed using a Sciex Triple TOF 4600 spectrometer in negative ion mode. GC-MS was performed on an Agilent 7890A / 5977A GC / MSD system using an Agilent HP-5MS column (30 m × 0.25 mm × 0.25 μm). Semi-preparative HPLC was performed on an Agilent 1260infinity system with a UV detector using a Boston Green ODS-AQ column (10 × 250 mm, 5 μm). Preparative HPLC was performed on a Shimadzu LC-20AP system using a YMC Actus Hydrosphere C18 column (30 × 250 mm, 5 μm) or a YMC Triart ODS column (50 × 250 mm, 7 μm). Column chromatography (CC) was performed using AB-8 resin (Sunresin NewMaterials Co., Ltd., China), MCI gel CHP-20 (Mitsubishi Chemical Systems, Inc., Japan), and Sephadex LH-20 (GE Healthcare Bio-Sciences AB, Sweden). Lyophilization was carried out on a Scientz-18N freeze dryer (Ningbo Scientz Biotechnology Co.). The chemical reagents are as follows: 95% EtOH, MeOH, MeCN, n-BuOH, and EtOAc, all AR grade (Cinc High Purity Solvents (Shanghai) Co., Ltd.); hydrochloric acid, NaOH, pyridine, and n-hexane, all AR grade (Sinopharm Chemical Reagent Co., Ltd.); acetonitrile for semi-preparative HPLC, HPLC grade (Merck KGaA); and L-cysteine ​​methyl ester hydrochloride, L-trimethylsilimidazole, D-glucose reference standard, and L-glucose reference standard, all LR grade (Shanghai Macklin Biochemical Co., Ltd.).

[0229] source:The dried whole plants of *Hesperidium rubrum* were collected from Anshun, Guizhou. The presence of the provisional hesperidin dihydrochalcone-4'-β-D-glucoside (HDCG) and six analogues was confirmed by LC-MS.

[0230] extract: The dried powder of *Smilax china* (10 kg) was extracted with 80% ethanol (100 L) at 70 °C for 4 h. After evaporation of the ethanol, the solution was concentrated to a small volume (5 L).

[0231] LC-MS-guided separation

[0232] The concentrated solution was subjected to an AB-8 resin column and eluted with 10% ethanol (100 L) and 95% ethanol (60 L). The 95% ethanol eluent was applied to an MCI column and gradually eluted with MeOH / H₂O (1:4 → 4:1) to provide five fractions. EICs targeting quasi-ions of HDCG and its possible analogues (e.g., [MH]) were then analyzed. - The fraction (m / z 465) was used for guiding fractionation and separation. Fraction 1 (70 g) was dissolved in water (3 L), the pH was adjusted to 1 with hydrochloric acid, and partitioned with EtOAc (3 L). The aqueous fraction (30 g) was adjusted to pH 7 with 1% NaOH solution and concentrated to 20 mL. The 20 mL sample was subjected to a Sephadex LH-20 column, followed by preparative HPLC (YMC Actus Hydrosphere C) at 30 mL / min in water-acetonitrile (74:26). 18 ,30×250mm,5μm), to produce CC-00487 (180mg).

[0233] Fraction 2 (60 g) was dissolved in water (3 L), and the pH was adjusted to 1 with hydrochloric acid. It was then extracted twice with EtOAc (3 L). The EtOAc fractions were combined and concentrated to obtain a semi-solid (20 g), which was dissolved in 50 mL of methanol-water (1:1) and subjected to chromatography on an MCI column and a Sephadex LH-20 column. Finally, it was purified by preparative HPLC at 30 mL / min on water-acetonitrile (79:21) to obtain CC-00486 (12 mg).

[0234] Part of 3 (20 g) was dissolved in 100 mL of methanol-water (7:3) and chromatographically analyzed on a Sephadex LH-20 column. The solution was further purified by preparative HPLC at 60 mL / min in water-acetonitrile (78:22) to obtain CC-00549 (3 mg).

[0235] Fraction 4 (60 g) was dissolved in 4% NaOH and the pH was adjusted to 10 with hydrochloric acid, followed by extraction with n-butanol (4 L). The aqueous phase was adjusted to pH 5 and partitioned again with n-butanol (4 L). The second n-butanol fraction (25 g) was subjected to chromatography on a Sephadex LH-20, eluted with 50% ethanol, to produce a mixture containing the target mass of the two fractions. Mixture 1 was further purified by semi-preparative HPLC using a Boston Green ODS-AQ (10 × 250 mm, 5 μm) at 3.5 mL / min water-acetonitrile (74:26) to produce hesperidin dihydrochalcone-4'-β-D-glucoside (HDCG, 2.3 mg) and CC-00474 (250 mg). Mixture 2 was further purified by preparative HPLC at 60 mL / min water-acetonitrile (72:28) to give compounds CC-00536 (34 mg) and CC-00537 (16 mg).

[0236] Determination of the absolute configuration of glucose

[0237] 2.4 mg CC-00486 and 2.6 mg CC-00487 were dissolved separately in 1 M HCl water (5 mL). Each solution was heated to reflux at 100 °C for 1.5 h. The mixture was cooled to room temperature and extracted twice with EtOAc (5 mL / time). The aqueous layer was evaporated to dryness under reduced pressure to provide glucose residue. The residue / D-glucose / L-glucose were dissolved separately in pyridine (0.5 mL), and 0.5 mL of L-cysteine ​​methyl ester hydrochloride was added. The mixture was maintained at 60 °C for 2 h, dried under reduced pressure, and alkylated with 1-trimethylsilimidazole (0.5 mL) trimethylsilylmethyl silylyl alkylate for 2 h. The mixture was partitioned between hexane and water (1.0 mL each), and the hexane extract was analyzed by GC-MS under the following conditions: column temperature, 230 °C; injection temperature, 250 °C; support, He gas; split ratio, 10:1; flow rate, 0.8 mL / min; MS scan range, 50–500.

[0238] result

[0239] HDCG was obtained as a white amorphous powder and exhibited properties as determined by HRESIMS analysis ([MH]). - The molecular formula C (m / z 465.1422, calculated value 465.1412, Δ = 2.1ppm) is... 22 H 26 O 11A single unit loss of 162 Daltons at m / z 303.0873 observed by MS / MS, along with an antecedent (δ 4.85, d, J = 7.7 Hz, H⁻¹”), indicates the presence of a β-glucose moiety in the structure. Its aglycone... 1 1H NMR spectra (Table 1) and 3-hydroxyphlorizin-4'-O-β-D-glucoside 1 The HNMR spectra are similar, except that the hydroxyl groups are methoxyl instead of hydroxyl groups, indicating the presence of a phloroglucinol ring [δ 6.12 (2H, s, H-3' and H-5')] and a catechol ring [δ 6.65 (d, J = 1.8 Hz, H-2), 6.79 (d, J = 8.2 Hz, H-5) and 6.59 (dd, J = 1.8, 8.2 Hz, H-6)].

[0240] Table 1 NMR spectral data of HDCG and CC-00474 a

[0241]

[0242] a NMR data were recorded on a Bruker Avance 600 in DMSO-d6. 1 H: 600MHz 13 C: 150MHz

[0243] exist 1 H- 1 In the H COSY experiment, the two methylene groups were coupled [δ 3.26 (2H,t,J = 7.6 Hz, H-8) and 2.74 (2H,t,J = 7.6 Hz, H-7)]. The key ROESY correlation of OCH3 / H-5 anchored the methoxy group at C-4, which was further determined by the correlation network of OCH3 / C-4, H-2 / C-4, and H-6 / C-4 in the HMBC experiment. Similarly, the glucosyl unit was linked to C-4' via the H-1” / C-4’ correlation. In summary, HDCG was identified as the known hesperidin dihydrochalcone-4'-O-β-D-glucan. The key correlations of HDCG are shown below:

[0244]

[0245] CC-00474 was obtained as a white amorphous powder and had the same molecular formula as HDCG by HRESIMS analysis ([MH]-, m / z 465.1408, calculated 465.1402, Δ = 1.3 ppm). 1 H-NMR and 13A comprehensive comparative analysis of the C-NMR spectra (Table 1) and HDCG showed that the only difference was the position of the methoxy and hydroxyl groups in the catechol ring. Cross-peaks of OCH3 / H-2 in the ROESY experiment indicated that the methoxy group was attached to C-3 instead of C-4, further confirmed by the correlation network of OCH3 / C-3 and H-5 / C-3 in the HMBC experiment. Based on the above evidence, CC-00474 was identified as a novel compound and named 3-methoxyphodermatin-4'-O-β-D-glucoside. The key correlations of CC-00474 are shown below:

[0246]

[0247]

[0248] CC-00486 was obtained as a white amorphous powder and showed promise as determined by HRESIMS analysis ([MH]). - The compound has the same molecular formula as 3-hydroxyphloretin-2'-O-β-D-glucoside (m / z 465.1414, calculated value 465.1402, Δ = 2.6 ppm). The 1H NMR spectrum of the compound (Table 2) is similar to that of 3-hydroxyphloretin-2'-O-β-D-glucoside, except that it is methoxy instead of hydroxyl, which indicates the presence of a phloroglucinol ring [δ 6.15 (1H, d, J = 1.7 Hz, H-3') and 5.92 (1H, d, J = 1.7 Hz, H-5')] and a catechol ring [δ 6.80 (d, J = 1.2 Hz, H-2), 6.65 (d, J = 8.0 Hz, H-5) and 6.62 (dd, J = 1.3, 8.0 Hz, H-6)].

[0249] Table 2 NMR spectral data of CC-00486 and CC-00487 a

[0250]

[0251]

[0252] a NMR data were recorded on a Bruker Avance 600 in DMSO-d6. 1 H: 600MHz 13 C: 150MHz

[0253] exist 1 H- 1In the H COSY experiment, the two methylene groups were coupled [δ 3.43 (1H, m, H-8), 3.34 (1H, m, H-8), and 2.80 (2H, t, J = 8.0 Hz, H-7)]. The key ROESY correlation of OCH3 / H-2 anchored the methoxy group at C-3, further confirmed by the correlation network of OCH3 / C-3, H-2 / C-4, and H-5 / C-3 in the HMBC experiment. Similarly, the correlations of H-1” / H-3’ in the ROESY experiment and H-1” / C-2’ in the HMBC experiment showed that the glucose unit was attached to C-2’. The absolute configuration of the glucose moiety was determined to be D-configuration using the above methods. Based on the above evidence, CC-00486 was identified as a novel compound, 3-methoxyphodermatin-2’-O-β-D-glucan. The key correlations of CC-00486 are shown below:

[0254]

[0255] CC-00487 was obtained as a white amorphous powder. Its HRESIMS peaks show a quasi-molecular ion peak at m / z 627.1931 [MH]. - The calculated value is 627.1931, Δ = 0.1 ppm, which is consistent with the molecular formula C. 28 H 36 O 16 . 1 The 1H NMR spectra (Table 2) show two sets of signals from the sugar fraction, and the coupling constants of the sugar fraction protons [(δ5.01, d, J = 7.4 Hz, H⁻¹”) and (δ4.96, d, J = 7.4 Hz, H⁻¹”’)] indicate the presence of two β-linked glucose molecules. Additionally, the non-sugar fraction... 1 H NMR spectrum and CC-00486 1 The similar 1H NMR spectra indicate the presence of a pyrogallol ring [δ 6.35, d, J = 2.3 Hz, H-3' and δ 6.16, d, J = 2.3 Hz, H-5'] and a catechol ring [δ 6.79 (d, J = 1.6 Hz, H-2), 6.65 (d, J = 8.0 Hz, H-5) and 6.62 (dd, J = 1.7, 8.0 Hz, H-6)]. 1 H- 1In the H COSY experiment, the two methylene groups were coupled [δ 3.45 (1H,m,H-8), 3.32 (1H,m,H-8), and 2.81 (2H,t,J=7.5Hz,H-7)]. The key ROESY correlation of OCH3 / H-2 anchored the methoxy group at C-3, further confirmed by the correlation network of OCH3 / C-3, H-2 / C-4, and H-5 / C-3 in the HMBC experiment. Similarly, in the HMBC experiment, the correlation of H-1”' / C-4' showed one glucose unit attached to C-4', and the correlation of H-1” / C-2' showed another attached to C-2'. Using the above methods, the absolute configuration of the glucose moiety was determined to be D-configuration. Based on the above evidence, CC-00487 was identified as a novel compound, 3-methoxyphodermatin-2',4'-O-β-D-glucan. The key correlations are shown below:

[0256]

[0257] CC-00549 was obtained as a brown amorphous powder with the molecular formula C. 23 H 26 O 13 The results were derived from the negative ion mode HRESIMS (m / z 509.1314 [MH]-, calculated value 509.1301, Δ = 2.6 ppm). ¹H-NMR data (Table 3) show a set of ABX-type aromatic H atom signals at δ 6.70 (¹H, d, overlap, H-2), 6.71 (¹H, d, J = 8.0 Hz, H-5), and 6.56 (¹H, dd, J = 8.0, 1.5 Hz, H-6), a singlet in the high-field aromatic region at δ 6.30 (¹H, s, H-5'), and two coupled methylene signals at δ 3.34 (2H, m, H-8) and 2.80 (2H, t, J = 8.0 Hz, H-7). The data indicate the presence of a dihydrochalcone skeleton and a penta-substituted B ring in the molecule. Furthermore, a pyranose syl unit (anomeric H and C atom signals, at δ 4.94 (1H, d, overlap, H-1”) and δ 101.4 (C-1”) respectively) was obtained from CC-00549. 1 H-NMR and 13 C-NMR data confirm this.

[0258] Table 3 NMR spectral data of CC-00549, CC-00536 and CC-00537 a

[0259]

[0260]

[0261] a NMR data were recorded on a Bruker Avance III 500 in menthol-d4. 1 H: 500MHz 13 C: 125MHz

[0262] HMBC correlations of the glycosyl group H-1” / C-4’ and ROESY correlations between the glycosyl groups H-1” and H-5’ confirmed the position of the pyranose glycosyl group at C-4’. Furthermore, in the HMBC experiments, long-range correlations of H-7’ / C-2’, C-3’, C-4’, and the carboxyl group C-8’ showed that the methylene C-7’ was linked to C-3’. Key correlations are shown below:

[0263]

[0264] CC-00536 was obtained as a pale yellow amorphous powder with the molecular formula C. 28 H 28 O 15 The negative ion HRESIMS has a m / z of 603.1354 ([MH)). - The peak at the calculated value of 603.1355 (Δ = -0.2 ppm) is specified. 1 H-NMR (500MHz, methanol-d4) and 13 C-NMR (125 MHz, methanol-d4, Table 3) allowed CC-00536 to be identified as the known 3-hydroxyphloretin 4'-O-[6”-O-galloyl]-β-D-glucopyranoside.

[0265] CC-00537 was obtained as a pale green amorphous powder. It has the same molecular formula as CC-00536 by HRESIMS analysis (m / z 603.1377 ([MH] - calculated value 603.1355, Δ = 3.6 ppm)). CC-00537 (Table 3) and CC-00536 have the same skeleton because they are very similar in NMR data, except for the difference in the glycosyl unit. Based on the lower field shift of H-4” [δ5.04(m)] and the higher field shift of H-6’ [δ3.70(d, J=12.2Hz), 3.59(m)] compared to CC-00536, the position of the galloyl group in CC-00537 was determined to be at C-4” of glucose instead of C-6”. This is also supported by the long-range correlation (δ167.7, C-7”’) between H-4” and carbonyl group in the galloyl unit in the HMBC experiment. Therefore, CC-00537 is deduced as a new compound, 3-hydroxyphloretin 4’-O-[4’’-O-galloyl]-β-D-glucopyranoside. The key correlations are shown below:

[0266]

[0267]

[0268] Example 2: Enhanced sweetness

[0269] Prepare a sample with the following components:

[0270] Table 1: 2% CC-00474 in PG

[0271] CC-00474 2mg Propylene glycol 98mg

[0272] Table 2: Rebaudioside M at 250 ppm in acidic water

[0273] Lebodiin M 125mg water 499.75g Citric acid 125mg

[0274] Table 3: 20 ppm CC-00474 and 250 ppm Rebaudioside M in acidic water

[0275] 250 ppm of rebaudioside M in acidic water (Table 2) 100g CC-00474 in PG (Table 1) 0.1g

[0276] Table 4: Rebaudioside M and PG at 250 ppm in acidic water (control)

[0277] 250 ppm of rebaudioside M in acidic water (Table 2) 100g PG 98mg

[0278] CC-00474 in PG was prepared in a weighing boat and added to a Reb M solution with stirring. The weighing boat was rinsed with Reb M solution. The solution was stirred until it was visually clear, then the sample was poured into a glass bottle and stored at 4°C.

[0279] Remove the bottle from the refrigerator and pour approximately 25 ml of the beverage into a 4-ounce plastic cup. Before tasting and between different samples, give the expert team members mineral water to rinse their mouths. Before tasting the next sample, give the expert team members unsalted biscuits to eat, followed by rinsing their mouths with mineral water.

[0280] Samples of rebaudioside M in acidic water, as well as samples of CC-00474 and rebaudioside M in acidic water, were given to the panel members. Panel members were asked to evaluate the sweetness of the samples and describe any differences in taste characteristics. Panel members were instructed to sip the samples, evaluate the sweetness, and then spit the samples into cups provided for this purpose.

[0281] All members of the expert panel agreed that the sample containing CC-00474 and rebaudioside M was sweeter than the sample containing only rebaudioside M.

Claims

1. A method for separating at least one dihydrochalcone compound, the method comprising: (a) Providing a crude extract of *Hemiberlesia lataniae* plant material, wherein the plant material is completely dried prior to extraction and subsequently extracted with a solvent consisting of 70% to 90% of one or more alcohols and the balance water, wherein the one or more alcohols are selected from the group consisting of methanol, ethanol, n-propanol, 2-propanol, 1-butanol and 2-butanol, and the extraction is carried out at a temperature of 60°C to 80°C for 1 hour to 24 hours; (b) Passing the crude extract through a column packed with polymer adsorption resin and eluting with an aqueous ethanol solvent containing at least 75% ethanol to provide a first solution containing at least one dihydrochalcone compound; and (c) Purify the first solution by reversed-phase chromatography using methanol and water as eluents to provide a second solution containing at least one dihydrochalcone compound. The at least one dihydrochalcone compound is 3-methoxyphodermin-2',4'-O-β-D-glucoside, 3-methoxyphodermin-4'-O-β-D-glucoside, or 3-methoxyphodermin-2'- O-β- D-glucoside.

2. The method as described in claim 1, wherein, The at least one dihydrochalcone compound is 3-methoxyphodermin-2',4'-O-β-D-glucoside, and the method further comprises: (d) Dilute the second solution with water to provide a diluted solution; (e) By contacting the diluted solution with acid, the diluted solution is acidified to a pH of 1 to 3 to provide an acidified solution; (f) Dispensing the acidified solution with an organic solvent to provide an aqueous portion and an organic portion; (g) By contacting the aqueous portion with an alkali, the aqueous portion is alkalized to a pH of 6 to 8 to provide a solution containing 3-methoxyphodermatin-2',4'-O-β-D-glucoside; (h) Concentrate the solution containing 3-methoxyphodermin-2',4'-O-β-D-glucoside to provide a concentrated solution; and (i) The concentrated solution is purified to provide a composition comprising 3-methoxyphodermatin-2',4'-O-β-D-glucoside having a purity of at least 80% by weight.

3. The method as described in claim 1, wherein, The at least one dihydrochalcone compound is 3-methoxyphodermin-4'-O-β-D-glucoside, and the method further comprises: (d) Dilute the second solution with an aqueous base to provide a diluted solution; (e) Acidifying the diluted solution to a pH of 8 to 10 by contacting it with acid to provide an acidified solution; (f) Dispensing the acidified solution with an organic solvent to provide an aqueous portion and an organic portion; (g) By contacting the aqueous portion with an acid, the aqueous portion is acidified to a pH of 4 to 6 to provide a second acidified solution; (h) Dispense the second acidified solution with an organic solvent to provide a second aqueous portion and a second organic portion; (i) concentrating the second organic fraction to provide a concentrated solution; and (j) Purify the concentrated solution to provide a composition comprising 3-methoxyphodermatin-4'-O-β-D-glucoside having a purity of at least 80% by weight.

4. The method of claim 1, wherein, The at least one dihydrochalcone compound is 3-methoxyphodermin-2'- O-β- D-glucosinolate, and the method further includes: (d) Dilute the second solution with water to provide a diluted solution; (e) By contacting the diluted solution with acid, the diluted solution is acidified to a pH of 1 to 3 to provide an acidified solution; (f) Dispensing the acidified solution with an organic solvent to provide an aqueous portion and an organic portion; (g) Concentrating the organic fraction to provide a concentrated solution; and (h) Purify the concentrated solution to provide 3-methoxyphodermatin-2'- with a purity of at least 80% by weight. O-β- Compositions of D-glucosides.

5. The method according to claim 2, wherein the eluent consists of 20% methanol and water.

6. The method according to claim 4, wherein the eluent consists of 40% methanol and water.

7. The method according to claim 3, wherein the eluent is composed of 60% methanol and water.

8. The method of claim 2, wherein the organic solvent used in the dispensing step is ethyl acetate.

9. The method of claim 4, wherein the organic solvent used in the dispensing step is ethyl acetate.

10. The method of claim 3, wherein the organic solvent used in the dispensing step is n-butanol.

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