Compositions

Abstract

To provide alternative and / or improved sweetness modifying composition.SOLUTION: There is provided a sweetness modifying composition comprising: one or more high-intensity sweetener(s) selected from steviol glycosides and / or mogrosides; and one or more low-intensity sweetener(s) selected from cellobiose, psicose, cyclamate and / or 11-O-mogroside V, wherein the sweetness modifying composition increases the sweetness of a sweetened composition by more than the sweetness of the sweetness modifying composition alone; and / or wherein the ratio of the one or more high-intensity sweetener(s) to the one or more low-potency sweetener(s) ranges from about 2:1 to about 12:1.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention generally relates to the use of one or more low-potency sweetener(s) to improve one or more sweetness characteristics of one or more high-intensity sweetener(s). Accordingly, the present invention also relates to compositions comprising a mixture of at least one high-intensity sweetener and at least one low-potency sweetener. The present invention further relates to the use of a combination of at least one high-intensity sweetener and at least one low-potency sweetener as a sweetness modifier when used in combination with at least one other sweetener and / or as a sweetener. The present invention further relates to the use of one or more mogroside(s) as a sweetness enhancer in sweetened compositions, and to said sweetened compositions. The present invention further relates to methods of making the sweeteners and compositions disclosed herein. [Background technology]

[0002] Sweetness is often a desirable characteristic in edible products, which are products intended to be taken orally, either for permanent ingestion or for temporary expulsion. Traditionally, sweetness has been provided by the addition of one or more sweeteners, particularly low-potency nutritive sweeteners such as sucrose (table sugar), fructose, glucose, xylose, arabinose, and rhamnose; sugar alcohols such as erythritol, xylitol, mannitol, sorbitol, and inositol; and sugar syrups such as high-fructose corn syrup and starch syrup. These impart a significant sweetness without any undesirable aftertaste. However, to reduce the caloric value of edible products, it is desirable to use reduced amounts of these sweeteners. Therefore, it is desirable to provide alternative sweeteners that can reduce the caloric value of edible products while maintaining the same or similar sweet taste.

[0003] High-intensity sweeteners (HIS) have been used for this purpose. High-intensity sweeteners can be natural or artificial and have a sweetness that can be hundreds of times that of sucrose, and therefore, in theory, can replace much larger amounts of sugar in a composition. Examples of high-intensity sweeteners include sucralose, saccharin, aspartame, acesulfame potassium (AceK), neotame, advantame, steviol glycosides (including stevioside, rebaudioside A, and rebaudioside D), or steviol glycoside mixture preparations having rebaudioside A and / or stevioside as the predominant component. However, these substances generally have the disadvantage that they can impart an undesirable off-taste, typically a bitter, metallic, or licorice-like taste, or an undesirable lingering sweetness to edible products. It would therefore be desirable to provide alternative and / or improved sweet taste altering compositions and sweetened compositions to address one or more of these problems. Summary of the Invention

[0004] According to a first aspect of the present invention, one or more high intensity sweetener(s) selected from the group consisting of steviol glycosides and / or mogrosides; and 1. A sweet taste modifying composition comprising one or more low intensity sweetener(s) selected from the group consisting of cellobiose, psicose, cyclamate and / or 11-O-mogroside V; wherein the sweet taste modifying composition increases the sweetness of the sweetened composition to a greater extent than the sweetness of the sweet taste modifying composition alone; and / or wherein the ratio of one or more high-intensity sweetener(s) to one or more low-potency sweetener(s) ranges from about 2:1 to about 12:1; The sweet taste altering compositions are provided.

[0005] According to a second aspect of the present invention, at least one sweetener present in an amount having a sweetness equal to or greater than about 1.5% (w / v) sucrose equivalent; and Sweet taste modifying composition according to any aspect or embodiment of the present invention A sweetened composition is provided comprising:

[0006] According to a third aspect of the present invention, there is provided use of one or more low-potency sweetener(s) selected from the group consisting of cellobiose, psicose, cyclamate, and / or 11-O-mogroside V to improve one or more sweetness characteristics of a sweetened composition comprising one or more high-intensity sweetener(s) selected from the group consisting of steviol glycosides and / or mogrosides, wherein the combined concentration of the one or more low-potency sweetener(s) and one or more high-potency sweetener(s) used has a sweetness of less than 1.5% (w / v) sucrose equivalent.

[0007] According to a fourth aspect of the present invention, there is provided a method for enhancing the sweetness of a sweetened composition, the method comprising providing a base composition comprising at least one sweetener present in an amount at or above its sweetness perception threshold and / or in an amount having a sweetness equivalent to or greater than about 1.5% (w / v) sucrose equivalent, and adding one or more high-intensity sweetener(s) selected from the group consisting of steviol glycosides and / or mogrosides, and one or more low-potency sweetener(s) selected from the group consisting of cellobiose, psicose, cyclamate and / or 11-O-mogroside V, wherein the ratio of one or more high-intensity sweetener(s) to one or more low-potency sweetener(s) is from about 2:1 to about 12:1; and / or wherein one or more high-intensity sweetener(s) are added in a total amount equal to or greater than about 15 ppm, and optionally equal to or less than about 50 ppm, and one or more low-potency sweetener(s) are added in a total amount equal to or greater than about 2 ppm, and optionally equal to or less than about 12 ppm; and / or wherein the combined concentration of the one or more high-intensity sweetener(s) and one or more low-potency sweetener(s) added has a sweetness of less than 1.5% (w / v) sucrose equivalent.

[0008] According to a fifth aspect of the present invention, there is provided a method of making a sweetened composition according to any aspect or embodiment of the present invention, the method comprising combining one or more high-intensity sweetener(s) with one or more low-potency sweetener(s). According to a sixth aspect of the present invention, there is provided a method of making a sweetened composition according to any aspect or embodiment of the present invention, the method comprising combining a base composition, one or more high-intensity sweetener(s), one or more low-intensity sweetener(s), and at least one other sweetener.

[0009] According to a seventh aspect of the present invention, there is provided a sweetened composition comprising at least one sweetener present in an amount having a sweetness equal to or greater than 1.5% (w / v) sucrose equivalent; and one or more sweetness enhancers selected from mogroside IV, siamenoside, and neomogroside. According to an eighth aspect of the present invention, there is provided the use of one or more of mogroside IV, siamenoside and neomogroside for enhancing the sweetness of a sweetened composition. Thus, in a further aspect, there is provided a method for enhancing the sweetness of a sweetened composition, the method comprising providing a base composition and adding at least one sweetener and one or more sweetness enhancers selected from mogroside IV, siamenoside and neomogroside.

[0010] According to a ninth aspect of the present invention, there is provided a method of making a sweetened composition according to any aspect or embodiment of the present invention, the method comprising combining a base composition with one or more sweetness enhancers selected from mogroside IV, siamenoside and neomogroside, and at least one other sweetener. According to a tenth aspect of the present invention, there is provided a sweetened composition comprising one or more mogroside(s). The one or more mogroside(s) may, for example, be present as a sweetness enhancer and thus may be present in an amount having a sweetness less than 1.5% (w / v) sucrose equivalent. The sweetened composition then further comprises at least one sweetener present in an amount having a sweetness equal to or greater than 1.5% (w / v) sucrose equivalent.

[0011] According to an eleventh aspect of the present invention, there is provided the use of one or more mogroside(s) for enhancing the sweetness of a sweetened composition. Thus, there is provided a method for enhancing the sweetness of a sweetened composition, comprising providing a base composition and adding at least one sweetener and one or more mogroside(s). According to a twelfth aspect of the present invention, there is provided a method of making a sweetened composition according to any aspect or embodiment of the present invention, said method comprising combining a base composition, one or more mogroside(s), and at least one other sweetener.

[0012] In some embodiments of any of the aspects of the invention, one or more high-intensity sweeteners may be or include mogroside V and / or one or more low-potency sweeteners may be or include 11-O-mogroside V. In some embodiments of any of the aspects of the invention, the ratio of one or more high intensity sweetener(s) to one or more low potency sweetener(s) is from about 2:1 to about 12:1. In some embodiments of any of the aspects of the invention, the ratio of one or more high intensity sweetener(s) to one or more low potency sweetener(s) is from about 5:1 to about 12:1. In some embodiments of any of the aspects of the invention, the ratio of one or more high intensity sweetener(s) to one or more low potency sweetener(s) may be from about 6:1 to about 10:1.

[0013] In some embodiments of any of the aspects of the invention, one or more high-intensity sweetener(s) may be present in a total amount ranging from about 15 ppm to about 30 ppm, and / or one or more low-potency sweetener(s) may be present in a total amount ranging from about 2 ppm to about 10 ppm. In some embodiments of any of the aspects of the invention, one or more high-intensity sweetener(s) may be present in a total amount ranging from about 22 ppm to about 28 ppm, and / or one or more low-potency sweetener(s) may be present in a total amount ranging from about 2 ppm to about 5 ppm.

[0014] In certain embodiments of the seventh through twelfth aspects of the present invention, the one or more mogroside(s) or one or more sweetener(s) may be present in an amount ranging from about 15 ppm to about 50 ppm, hi certain embodiments, the one or more mogroside(s) or one or more sweetener(s) may be present in an amount ranging from about 15 ppm to about 35 ppm. In some embodiments of any aspect of the present invention, the combination of one or more high-intensity sweetener(s) and one or more low-potency sweetener(s) alone may have a sweetness of less than about 1.5% (w / v) sucrose equivalent. Specifically, the concentration of one or more high-intensity sweetener(s) and one or more low-intensity sweetener(s) in the sweetened composition may have a sweetness of less than about 1.5% (w / v) sucrose equivalent.

[0015] In certain embodiments of the seventh through twelfth aspects of the present invention, the one or more mogroside(s) or one or more sweetness enhancer(s) may have a total sweetness of less than about 1.5% (w / v) sucrose equivalent, hi certain embodiments, the one or more sweetness enhancers increase the sweetness of the sweetened composition by more than the total sweetness of the one or more sweetness enhancers alone.

[0016] In some embodiments of any aspect of the invention, the combination of one or more high-intensity sweetener(s) with one or more low-potency sweetener(s) can increase the sweetness of the sweetened composition by a greater amount than the sweetness of the combination alone. In some embodiments of any aspect of the invention, the combination of one or more high-intensity sweetener(s) with one or more low-potency sweetener(s) can increase the sweetness of the composition by an amount equal to or greater than about 1.25% (w / v) sucrose equivalent. In some embodiments of any aspect of the invention, the one or more low-potency sweetener(s) attenuate the lingering sweet taste of a sweetened composition that includes one or more high-intensity sweetener(s) compared to the lingering sweet taste of the sweetened composition in the complete absence of the one or more low-potency sweetener(s).

[0017] In some embodiments of any aspect of the invention, the one or more low-potency sweetener(s) attenuate the bitter and / or astringent taste of a sweetened composition that includes one or more high-intensity sweetener(s) compared to the bitter and / or astringent taste of the sweetened composition in the complete absence of the one or more low-potency sweetener(s). One or more (e.g., all) of the sweeteners used can be natural or synthetic (artificial). One or more of the sweeteners can be made, for example, by a biological process, an enzymatic process, or a synthetic process.

[0018] Certain embodiments of any aspect of the present invention may provide one or more of the following advantages: Increased sweetness in the composition; Enhanced sweetness in compositions including at least one sweetener; · A reduction in the amount of caloric sweetener needed to achieve the desired sweetness; · Improvement of one or more sweetness attributes to make the sweet taste more similar to sugar (sucrose); · Reduced lingering sweetness (e.g., reducing the length of time the sweet taste lingers and / or reducing the intensity of the sweet taste more quickly); Reduced bitter and / or astringent and / or licorice-like and / or metallic taste; Improvement in sweetness impact (e.g., increasing the maximum intensity of sweet taste and / or decreasing the length of time it takes for sweet taste to be detected) (e.g., decreasing lingering sweetness).

[0019] The details, examples, and preferences provided in connection with one or more specific aspects of any of the described aspects of the invention are described further herein and apply equally to all aspects of the invention. Any combination of the embodiments, examples, and preferences described herein in all possible variations is covered by the present invention unless otherwise indicated herein or otherwise clearly contradicted by context. [Brief explanation of the drawings]

[0020] [Figure 1] FIG. 1 shows the chromatogram of the Monk Fruit extract (Extract 2 in Table 1 below). [Figure 2] Figure 2 shows the chemical structures of mogrosides 1–6; [Figure 3A] Figure 3 shows the LS-MS analysis of commercially available Luo Han Guo extract; [Figure 3B] Figure 3 shows the LS-MS analysis of commercially available Luo Han Guo extract;

[0021] [Figure 4-1] Figure 4 shows the heteronuclear single quantum coherence-total correlation spectroscopy (HSQC-TOCSY) (hsqcgpmlph) of iso-mogroside VI at different mixing times (d9). A: 10 ms mixing time. B: 30 ms mixing time. C: 60 ms mixing time. D: 100 ms mixing time. Due to the overlap between H-1 of Glc II and H-6a of Glc III, the HSQC-TOCSY correlation intensity of Glc II was not analyzed here; [Figure 4-2]Figure 4 shows the heteronuclear single quantum coherence-total correlation spectroscopy (HSQC-TOCSY) (hsqcgpmlph) of iso-mogroside VI at different mixing times (d9). A: 10 ms mixing time. B: 30 ms mixing time. C: 60 ms mixing time. D: 100 ms mixing time. Due to the overlap between H-1 of Glc II and H-6a of Glc III, the HSQC-TOCSY correlation intensity of Glc II was not analyzed here; [Figure 5] Figure 5 shows the HSQC-TOCSY (hsqcgpmlph) peak intensity quantification of iso-mogroside VI glucopyranosyl at different mixing times. (*C-3 and C-5 signals on HSQC-TOCSY appeared overlapped for a mixing time of 100 ms. Therefore, the total integrals of C-3 and C-5 were used in the bar graphs);

[0022] [Figure 6]Figure 6 shows the strategy for elucidating mogroside glycans. (*The number of C-2–C-6 bonds appearing under a certain mixing time may vary slightly if the HSQC-TOCSY peak intensities are adjusted. Nevertheless, the linkage order can be determined by observing the increasing intensities of C-2–C-6 bonds in experiments with different mixing times. **So far, there has been no natural glycosylation on C-3 of mogroside glucopyranosyl. C-3 glycosylation on glucopyranosyl would cause a downshift from δ7 to δ81, which can be easily determined by HSQC-TOCSY experiments.) The sequence of steps in Figure 6 can be outlined as follows: In step 1, heteronuclear multiple bond correlation spectroscopy (HMBC) was used to determine the anomers C-1 and H-1 of the sugar. Starting from the linkage of the sugar to the aglycone, In step 2, HSQC-TOCSY was used with a mixing time of 100 ms to determine the entire group of C-2 through C-6. HSQC-TOCSY or HSQC-TOCSY (d9 = 10 ms) to assign C-2; HSQC-TOCSY (d9 = 30 ms) to assign C-3; HSQC-TOCSY (d9 = 60 ms) to assign C-4; and HSQC-TOCSY (d9 = 100 ms) to assign C-5 and C-6. In step 3, if a C-2 downshift from ~δ75 to ~δ81, a C-4 downshift from ~δ71 to ~δ81, or a C-6 downshift from ~δ62 to ~δ69 is observed, check HMBC for glycosylation at these positions. **If C-2 is downshifted from ~δ75 to ~δ81, C-4 is downshifted from ~δ71 to ~δ81, or C-6 is downshifted from ~δ62 to ~δ69, check HMBC for glycosylation at these positions.**

[0023] [Figure 7]Figure 7 shows the chemical structure for iso-mogroside VI, which has the chemical formula C66H112O34 and an exact mass of 1448.70. This chemical structure is referred to as Formula I; and [Figure 8] Figure 8 shows the chemical structure for 11-epi-mogroside V, which has the chemical formula CHO and an exact mass of 1286.65. This chemical structure is designated Formula II. DETAILED DESCRIPTION OF THE INVENTION

[0024] Detailed Description The present invention is based on the surprising discovery that a combination of one or more high-intensity sweetener(s) (e.g., mogroside V) and one or more low-potency sweetener(s) (e.g., 11-O-mogroside V) can act synergistically with at least one other sweetener (e.g., sucrose) to provide a composition having a sweetness that exceeds the sum of the sweetnesses of the individual sweeteners. The present invention is further based on the surprising discovery that one or more low-potency sweetener(s) offset one or more negative sweetness attributes of one or more high-potency sweetener(s). For example, a combination of one or more high-intensity sweetener(s) (e.g., mogroside V) with one or more low-potency sweetener(s) (e.g., 11-O-mogroside V) in a sweetened composition (i.e., a composition containing at least one other sweetener, such as sucrose, in an amount above its sweetness perception threshold and / or in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent) may provide an improved sweetness profile compared to the use of one or more high-intensity sweetener(s) alone. The sweetness profile may thus, for example, be closer to that of sucrose.

[0025] Thus, various compositions are provided herein that contain one or more high-intensity sweetener(s) and one or more low-potency sweetener(s) as disclosed herein, particularly sweetened compositions that contain at least one sweetener in an amount above its sweetness perception threshold and / or in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent, and one or more high-intensity sweetener(s) and one or more low-potency sweetener(s). Sweetened compositions may also be referred to as edible compositions. Various uses of one or more high-intensity sweetener(s) and one or more low-potency sweetener(s) as disclosed herein, as well as methods for making the various compositions disclosed herein, are also provided herein.

[0026] The present invention is further based on the surprising discovery that mogrosides, such as mogroside IV, siamenoside, and neomogroside, can act as sweetness enhancers (i.e., can increase the sweetness of a sweetened composition to a greater extent than the sweetness of the sweetness enhancer alone). Thus, provided herein are various compositions, particularly sweetened compositions, that include one or more of mogroside IV, siamenoside, and neomogroside.

[0027] composition Various compositions comprising at least one high-intensity sweetener and at least one low-potency sweetener are provided herein. Also provided herein are compositions comprising one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside. In some embodiments, the composition is an edible composition.

[0028] In some embodiments, a sweet taste modifying composition is provided that comprises, consists essentially of, or consists of at least one high-intensity sweetener selected from the group consisting of steviol glycosides and / or mogrosides, and at least one low-potency sweetener selected from the group consisting of cellobiose, psicose, cyclamate, and / or 11-O-mogroside V. In some embodiments, the sweet taste modifying composition comprises, consists essentially of, or consists of one high-intensity sweetener and one low-potency sweetener. The sweet taste modifying composition may be, for example, a concentrate that can be diluted in a sweetened (e.g., edible) composition to impart a desired sweetness to the edible composition. The term "sweetened composition" refers to a composition that contains at least one sweetener present in an amount above its sweetness perception threshold and / or in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent.

[0029] In some embodiments, a sweetened composition (e.g., an edible composition) is provided that includes one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside. In some embodiments, a sweetened composition (e.g., an edible composition) is provided that includes at least one high-intensity sweetener and at least one low-potency sweetener. The combination of high-intensity sweetener(s) and low-potency sweetener(s) may be referred to as a sweetness-modifying composition. One or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, may also be referred to herein as a sweetness-modifying composition. Thus, in some embodiments, sweetened compositions are provided, including a composition comprising at least one sweetener present in an amount above its sweetness perception threshold and / or in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent, and a sweet taste modifying composition comprising, consisting essentially of, or consisting of at least one high-intensity sweetener and at least one low-potency sweetener. The sweetened composition may be, for example, an edible composition.

[0030] The term "enhancing," when used in connection with a particular sweet taste modifier composition, refers to a synergistic sweetening effect when used in combination with at least one other sweetener. The sweet taste modifier composition increases the sweetness of the sweetened composition to a greater extent than the sweetness of the sweet taste modifier composition alone. In other words, the sweetness of a composition comprising at least one sweetener and at least one sweet taste modifier composition exceeds the sum of the sweetnesses of all the sweeteners in the composition. The sweet taste modifier compositions described herein are used in sweetened (e.g., edible) compositions in amounts that have no detectable sweetness or no perceived sweet taste (below their sweetness perception threshold). Typically, sweet taste modifier compositions with a sweetness below 1.5% (w / v) sucrose equivalent are recognized as "essentially non-sweet" by FEMA (Food and Flavor Manufacturers Association). Sweet taste modifiers may also be referred to as sweetness enhancers.

[0031] A sweetened composition comprising a sweet taste modifier composition as disclosed herein and at least one sweetener present in an amount above its sweetness perception threshold and / or in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent may have a sweetness greater than about 1.0% (w / v) sucrose equivalent or greater than the sweetness of the sweetened composition in the absence of the sweet taste modifier composition. For example, the sweetened composition may have a sweetness greater than about 1.1% (w / v) sucrose equivalent or greater, or about 1.15% (w / v) sucrose equivalent or greater, or about 1.2% (w / v) sucrose equivalent or greater, or about 1.25% (w / v) sucrose equivalent or greater than the sweetness of the sweetened composition in the absence of the sweet taste modifier composition. In other words, the sweet taste modifying composition can increase the sweetness of the sweetened composition by about 1% (w / v) sucrose equivalent or more, or about 1.1% (w / v) sucrose equivalent or more, or about 1.15% (w / v) sucrose equivalent or more, or about 1.2% (w / v) sucrose equivalent or more, or about 1.25% (w / v) sucrose equivalent or more. The comparative composition is identical to the sweet taste modifying composition except that it does not include the sweet taste modifying composition.

[0032] The term "sucrose equivalent" refers to the sweetness equivalence of a composition containing at least one non-sucrose sweetener relative to a reference sucrose solution. Typically, taste panelists are trained to detect the sweetness of a reference sucrose solution containing between 1% and 15% sucrose (w / v). Other non-sucrose sweeteners can then be tasted in a series of dilutions to determine the concentration of a non-sucrose sweetener that is as sweet (i.e., isosweet) relative to a given sucrose standard. The term "isosweet" refers to a composition having equivalent sweetness. Typically, the sweetness of a given composition is measured relative to a solution of sucrose. See "A Systematic Study of Concentration-Response Relationships of Sweeteners," G.E. DuBois, D.E. Walters, S.S. Schiffman, Z.S. Warwick, B.J. Booth, S.D. Pecore, K. Gibes, B.T. Carr, and L.M. Brands, in Sweeteners: Discovery, Molecular Design and Chemoreception, D.E. Walters, F.T. Orthoefer, and G.E. DuBois, Eds., American Chemical Society, Washington, D.C. (1991), pp. 261-276.

[0033] A combination of one or more high-intensity sweetener(s) and one or more low-potency sweetener(s) (e.g., a sweet taste modifier composition) may have a sweetness of less than about 1.5% (w / v) sucrose equivalent. For example, a combination of a high-intensity sweetener(s) and a low-potency sweetener(s) (e.g., a sweet taste modifier composition) may have a sweetness of less than or equal to about 1.45% (w / v) sucrose equivalent, or less than or equal to about 1.4% (w / v) sucrose equivalent, or less than or equal to about 1.35% (w / v) sucrose equivalent, or less than or equal to about 1.3% (w / v) sucrose equivalent. For example, a combination of high intensity sweetener(s) and low potency sweetener(s) (e.g., a sweet taste modifying composition) may have a sweetness equal to or greater than about 1% (w / v) sucrose equivalent, or equal to or greater than about 1.1% (w / v) sucrose equivalent, or equal to or greater than about 1.15% (w / v) sucrose equivalent, or equal to or greater than about 1.2% (w / v) sucrose equivalent, or equal to or greater than about 1.25% (w / v) sucrose equivalent, or equal to or greater than about 1.3% (w / v) sucrose equivalent.

[0034] The sweetness enhancer(s) selected from one or more mogroside(s), e.g., mogroside IV, siamenoside, and neomogroside, may have a sweetness of less than about 1.5% (w / v) sucrose equivalent. For example, the sweetness enhancer(s) selected from one or more mogroside(s), e.g., mogroside IV, siamenoside, and neomogroside, may have a sweetness equal to or less than about 1.45% (w / v) sucrose equivalent, or equal to or less than about 1.4% (w / v) sucrose equivalent, or equal to or less than about 1.35% (w / v) sucrose equivalent, or equal to or less than about 1.3% (w / v) sucrose equivalent. For example, one or more sweetness enhancers selected from one or more mogroside(s), e.g., mogroside IV, siamenoside, and neomogroside, may have a sweetness equal to or greater than about 1% (w / v) sucrose equivalent, or equal to or greater than about 1.1% (w / v) sucrose equivalent, or equal to or greater than about 1.15% (w / v) sucrose equivalent, or equal to or greater than about 1.2% (w / v) sucrose equivalent, or equal to or greater than about 1.25% (w / v) sucrose equivalent, or equal to or greater than about 1.3% (w / v) sucrose equivalent.

[0035] Each of the sweeteners and sweetness enhancers used in the compositions disclosed herein may be natural or synthetic (artificial) sweeteners. Examples of non-naturally occurring (i.e., synthetic) mogrosides are disclosed in WO 2017 / 075257, the contents of which are incorporated herein by reference. The term "natural sweetener" refers to a sweetener derived from nature, including mixtures that may have been enzymatically treated (e.g., glycosylated) to form compounds not found in nature (this does not include purified enzymatically treated compounds). For example, a modified extract having a mogrol glycoside distribution that differs from the naturally occurring mogrol glycoside distribution (e.g., enhanced) may be classified as natural. For example, glycosylated steviol glycosides and / or a mixture of glycosylated mogrosides may be classified as natural. Each of the sweeteners used in the compositions disclosed herein may be derived from food. A "food-derived" product refers to a product prepared under typical cooking conditions, such as using temperatures similar to those used in cooking methods. In some embodiments, the high-intensity sweetener and the low-potency sweetener used in the compositions disclosed herein (e.g., in the sweet taste-altering compositions disclosed herein) are both natural sweeteners. In some embodiments, all sweeteners used in the compositions disclosed herein are natural.

[0036] The sweeteners disclosed herein may be used in pure or purified form and may be chemically synthesized, produced by biotechnological processes (e.g., fermentation), or isolated from natural sources (e.g., plant sources including, but not limited to, fruits, sugar cane, sugar beets, etc.). One or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can be, for example, at least 80% pure by weight. For example, one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can be at least about 85% pure by weight, or at least about 90% pure by weight, or at least about 95% pure by weight, or at least about 98% pure by weight, or at least about 99% pure by weight. For example, one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can be up to 100% pure by weight, or up to 99% pure by weight.

[0037] The term "high-intensity sweetener" refers to a compound that is at least 100 times sweeter than sucrose. In some embodiments, a high-intensity sweetener is at least about 120, or at least about 140, or at least about 150, or at least about 160, or at least about 180, or at least about 200, or at least about 220, or at least about 240, or at least about 250, or at least about 260, or at least about 280, or at least about 300, or at least about 320, or at least about 340, or at least about 350, or at least about 360, or at least about 380, or at least about 400, or at least about 420, or at least about 440, or at least about 450 times sweeter than sucrose. A high-intensity sweetener may, for example, be up to at least 1000 times sweeter than sucrose. High intensity sweeteners have a sweetness at least 100 times that of sucrose, but in the context of their use in the sweet taste altering compositions as described herein, they will be used in an amount that does not have any detectable or perceivable sweetness in the sweetened composition (an amount that provides a sweetness of less than 1.5% (w / v) sucrose equivalent, recognized as "essentially unsweet" by FEMA (Food and Flavor Manufacturers Association)).

[0038] The one or more high-intensity sweetener(s) may be, for example, one or more steviol glycosides and / or one or more mogrosides. For example, the one or more high-intensity sweeteners may be a mixture of steviol glycosides and mogrosides. For example, the one or more high-intensity sweetener(s) may be one or more steviol glycosides. For example, the one or more high-intensity sweetener(s) may be one or more mogrosides. In some embodiments, mogrosides may perform better than steviol glycosides in terms of sweetness enhancement and off-note reduction (e.g., reducing lingering sweet aftertaste).

[0039] The high intensity sweetener can be, for example, one or more steviol glycoside(s). Examples of steviol glycosides include, for example, stevioside (CAS: 57817-89-7), rebaudioside A (CAS: 58543-16-1), rebaudioside B (CAS: 58543-17-2), rebaudioside C (CAS: 63550-99-2), rebaudioside D (CAS: 63279-13-0), rebaudioside E (CAS: 63279-14-1), rebaudioside F (CAS: 438045-89-7), rebaudioside G (CAS: 127345-21-5), rebaudioside H, rebaudioside I (CAS: 127345-21-5), rebaudioside II (CAS: 127345-21-5), rebaudioside III (CAS: 127345-21-5), rebaudioside B (CAS: 127345-21-5), rebaudioside C (CAS: 127345-21-5), rebaudioside D (CAS: 127345-21-5), rebaudioside E (CAS: 127345-21-5), rebaudioside F (CAS: 127345-21-5), rebaudioside H (CAS: 127345-21-5), rebaudioside I ... Rebaudioside I (CAS: 1220616-34-1), Rebaudioside J, Rebaudioside K, Rebaudioside L, Rebaudioside M (CAS: 1220616-44-3), Rebaudioside N (CAS: 1220616-46-5), Rebaudioside O (CAS: 1220616-48-7), Dulcoside A (CAS: 64432-06-0), Dulcoside B (CAS: 63550-99-2), Rubusoside (CAS: 64849-39-4), and Naringin Dihydrochalcone (CAS: 18916-17-1).

[0040] The high intensity sweetener can be, for example, one or more mogroside(s). In some embodiments, the high-intensity sweetener may be one or more of the mogrosides listed herein. In some embodiments, the high-intensity sweetener may be one or more of mogroside IV, siamenoside, neomogroside, and mogroside V (including all isomers thereof). For example, the high-intensity sweetener may be a mixture of mogroside IV, siamenoside, and mogroside V (including all isomers thereof). The one or more mogroside(s) may be obtained or obtainable from, for example, a swingle fruit extract.

[0041] The term "low-potency sweetener" refers to a compound that has a sweetness that is less than 100 times that of sucrose. In some embodiments, a low-potency sweetener has a sweetness that is up to about 95 times, or up to about 90 times, or up to about 85 times that of sucrose. The one or more low-potency sweetener(s) are selected from one or more of cellobiose, psicose, cyclamate, and / or 11-O-mogroside V (CAS: 126105-11-1). For example, the one or more low-intensity sweetener(s) can be one or more of cellobiose, psicose, and 11-O-mogroside V.

[0042] In some embodiments, one or more high-intensity sweetener(s) are or include a high-intensity mogroside. In some embodiments, one or more low-potency sweetener(s) are or include a low-potency mogroside. In some embodiments, one or more high-intensity sweetener(s) are or include a high-intensity mogroside, and one or more low-potency sweetener(s) are or include a low-potency mogroside. In some embodiments, one or more high-intensity sweeteners are or include mogroside V. In some embodiments, one or more low-potency sweeteners are or include 11-O-mogroside V. In some embodiments, one or more high-intensity sweeteners are or include mogroside V and one or more low-potency sweeteners are or include 11-O-mogroside V.

[0043] Mogrosides are a group of triterpene glycosides obtained from the fruit of Siraitia grosvenorii (also known as arhat fruit, longevity fruit, or swingle fruit). Mogrosides make up approximately 1% of the flesh of the fresh fruit. Through extraction, a powder form containing up to 80% mogrosides can be obtained. The mogroside extracts consisted of grosvenolin II, grosvenolin I, 11-O-mogroside II(I), 11-O-mogroside II(II), 11-O-mogroside II(III), mogroside II(I), mogroside II(II), mogroside II(III), 11-dehydroxy-mogroside III, 11-O-mogroside III, mogroside III(I), mogroside III(II), mogroside IV(I) (siamenoside), mogroside III(III). Mogroside IV(II), mogroside IV(III), mogroside IV(IV), deoxymogroside V(I), deoxymogroside V(II), 11-O-mogroside V(I), mogroside V isomers, mogroside V, iso-mogroside V, 7-O-mogroside V, 11-O-mogroside VI, mogroside VI(I), mogroside VI(II), mogroside VI(III) (neomogroside), and mogroside VI(IV). The exact amount of mogroside V may vary depending on the maturity of the fruit and / or the extraction process used.

[0044] Mogroside(s) include both naturally occurring mogroside(s) and non-naturally occurring mogrosides. Examples of mogrosides include, for example, grosvenolin II, grosvenolin I, 11-O-mogroside II(I), 11-O-mogroside II(II), 11-O-mogroside II(III), mogroside II(I), mogroside II(II), mogroside II(III), 11-dehydroxy-mogroside III, 11-O-mogroside III, mogroside III(I), mogroside III(II), mogroside IIIe, mogroside IIIx, mogroside IV(I), ) (siamenoside), mogroside IV(II), mogroside IV(III), mogroside IV(IV), deoxymogroside V(I), deoxymogroside V(II), 11-O-mogroside V(I), mogroside V isomer, mogroside V, iso-mogroside V, 7-O-mogroside V, 11-O-mogroside VI, mogroside VI(I), mogroside VI(II), mogroside VI(III) (neomogroside), and mogroside VI(IV). The mogroside(s) may be, for example, obtained or obtainable from a swingle fruit extract.

[0045] Mogroside V (CAS: 88901-36-4) is a glycoside of a cucurbitane derivative and has the chemical formula C 60 H 102 O 29 and has the chemical structure shown below: Mogroside V can be found in certain plant extracts, such as extracts from the fruit Siraitia grosvenorii. Pure mogroside V has been found to be at least 400 times sweeter than sucrose. [ka]

[0046] Siamenoside (CAS: 126105-12-2) is a cucurbitan found in the fruit of Siraitia grosvenorii and has the following chemical structure: [ka]

[0047] Mogroside IV (CAS: 89590-95-4) is a triterpene heteroside found in the fruits of Siraitia grosvenorii and has the following chemical structure: [ka]

[0048] Neomogroside (CAS: 189307-15-1) is also a cucurbitane glycoside found in the fruit of Siraitia grosvenorii and has the following chemical structure: [ka]

[0049] 11-O-mogroside V (CAS: 126105-11-1) is derived from mogroside V and has the following chemical structure: It is also found in plant extracts, such as extracts from the fruit Siraitia grosvenorii. 11-O-mogroside V has been found to be approximately 84 times sweeter than sucrose. [ka]

[0050] The ratio of one or more high-intensity sweetener(s) to one or more low-potency sweetener(s) is equal to or greater than about 2: 1. For example, the ratio of one or more high-intensity sweetener(s) to one or more low-potency sweetener(s) can be equal to or greater than about 2.5: 1, or equal to or greater than about 3: 1, or equal to or greater than about 3.5: 1, or equal to or greater than about 4: 1, or equal to or greater than about 4.5: 1, or equal to or greater than about 5: 1, or equal to or greater than about 5.5: 1, or equal to or greater than about 6: 1, or equal to or greater than about 6.5: 1, or equal to or greater than about 7: 1, or equal to or greater than about 7.5: 1, or equal to or greater than about 8: 1. The ratio of high-intensity sweetener(s) to low-potency sweetener(s) is equal to or less than about 12: 1. For example, the ratio of one or more high-intensity sweetener(s) to one or more low-potency sweetener(s) can be equal to or less than about 11.5:1, or equal to or less than about 11:1, or equal to or less than about 10.5:1, or equal to or less than about 10:1, or equal to or less than about 9.5:1, or equal to or less than about 9:1, or equal to or less than about 8.5:1. For example, the ratio of one or more high intensity sweetener(s) to one or more low potency sweetener(s) can range from about 5:1 to about 11:1, or from about 6:1 to about 10:1, or from about 6.5:1 to about 9.5:1, or from about 7:1 to about 9:1, or from about 7.5:1 to about 8.5:1.

[0051] In some embodiments, the ratio of one or more high-intensity sweetener(s) to one or more low-potency sweetener(s) is from about 2:1 to about 12:1, or from about 4:1 to about 12:1, or from about 5:1 to about 12:1, or from about 6:1 to about 10:1, or from about 7:1 to about 9:1. The ratio may be by weight or by volume. The ratio applies only to the high-intensity sweetener(s) and low-potency sweeteners (high-intensity and low-potency sweeteners used in the sweetened composition in an amount below the sweetness perception threshold or having less than 1.5% (w / v) sucrose equivalent) in the sweet taste altering composition.

[0052] The one or more high-intensity sweetener(s) may be present in the composition in a total amount equal to or greater than about 15 ppm. For example, the one or more high-intensity sweetener(s) may be present in the composition in a total amount equal to or greater than about 16 ppm, or equal to or greater than about 17 ppm, or equal to or greater than about 18 ppm, or equal to or greater than about 19 ppm, or equal to or greater than about 20 ppm, or equal to or greater than about 21 ppm, or equal to or greater than about 22 ppm, or equal to or greater than about 23 ppm, or equal to or greater than about 24 ppm, or equal to or greater than about 25 ppm. For example, one or more high intensity sweetener(s) may be present in the composition in a total amount of less than or equal to about 50 ppm, or less than or equal to about 48 ppm, or less than or equal to about 46 ppm, or less than or equal to about 45 ppm, or less than or equal to about 44 ppm, or less than or equal to about 42 ppm, or less than or equal to about 40 ppm, or less than or equal to about 38 ppm, or less than or equal to about 36 ppm, or less than or equal to about 35 ppm, or less than or equal to about 34 ppm, or less than or equal to about 32 ppm, or less than or equal to about 30 ppm.

[0053] For example, one or more high-intensity sweetener(s) may be present in the composition in a total amount ranging from about 15 ppm to about 50 ppm, or from about 15 ppm to about 45 ppm, or from about 15 ppm to about 40 ppm, or from about 15 ppm to about 35 ppm, or from about 15 ppm to about 30 ppm. For example, one or more high-intensity sweetener(s) may be present in the composition in a total amount ranging from about 15 ppm to about 30 ppm, or from about 20 ppm to about 30 ppm, or from about 22 ppm to about 28 ppm, or from about 23 ppm to about 27 ppm, or from about 24 ppm to about 26 ppm. For example, one or more high-intensity sweetener(s) may be present in the composition in a total amount of about 20 ppm or about 25 ppm. The composition may be, for example, a sweetened composition having a sweetness above its sweetness perception threshold and / or comprising at least one sweetener in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent.

[0054] One or more low-potency sweetener(s) may be present in the composition in a total amount of about 2 ppm or more. For example, one or more low-potency sweetener(s) may be present in the composition in a total amount of about 3 ppm or more. For example, one or more low-potency sweetener(s) may be present in the composition in a total amount of about 12 ppm or less, or about 11 ppm or less, or about 10 ppm or less, or about 9 ppm or less, or about 8 ppm or less, or about 7 ppm or less, or about 6 ppm or less, or about 5 ppm. For example, one or more low-potency sweetener(s) may be present in the composition in a total amount ranging from about 2 ppm to about 12 ppm, or from about 2 ppm to about 10 ppm, or from about 2 ppm to about 5 ppm, e.g., in a total amount of about 3 ppm.

[0055] The composition may, for example, include at least one sweetener other than a combination of a high intensity sweetener and a low potency sweetener as disclosed herein (eg, a sweetness altering composition). This concentration range may be particularly suitable for liquid compositions such as beverages, or compositions that do not contain any protein or fat. In compositions with a base such as milk and yogurt, or other compositions that contain protein and / or fat, higher concentrations of one or more high-intensity sweetener(s) and one or more low-potency sweetener(s) may be used. For example, a concentration about 1.5 times higher than that used for liquid compositions or compositions that do not contain any protein or fat may be used. For example, a concentration about 1.5 to about 3 times higher than that used for liquid compositions or compositions that do not contain any protein or fat may be used.

[0056] Thus, for example, one or more high intensity sweetener(s) may be present in a composition (e.g., a composition having a base such as milk and yogurt, or other composition comprising protein and / or fat) in a total amount ranging from about 20 ppm to about 75 ppm, e.g., from about 22 ppm to about 74 ppm, or from about 24 ppm to about 72 ppm, or from about 25 ppm to about 70 ppm, or from about 26 ppm to about 68 ppm, or from about 28 ppm to about 66 ppm, or from about 30 ppm to about 65 ppm, or from about 30 ppm to about 60 ppm, or from about 30 ppm to about 55 ppm, or from about 30 ppm to about 50 ppm, or from about 30 ppm to about 45 ppm.

[0057] Thus, for example, one or more low-potency sweetener(s) may be present in a composition (e.g., a composition having a base such as milk and yogurt, or other composition containing protein and / or fat) in a total amount ranging from about 3 ppm to about 20 ppm, or from about 4 ppm to about 18 ppm, or from about 4 ppm to about 16 ppm, or from about 5 ppm to about 15 ppm, or from about 6 ppm to about 15 ppm.

[0058] In some embodiments, a sweetened composition comprises at least one sweetener having a sweetness above its sweetness perception threshold and / or in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent, and a sweet taste modifying composition as described herein consisting of 15 ppm to about 50 ppm of one or more high-intensity sweetener(s) as described herein and 2 ppm to 12 ppm of one or more low-potency sweetener(s) as described herein. In some embodiments, an edible composition comprises at least one sweetener and a sweet taste modifying composition consisting of 15 ppm to about 30 ppm of one or more high-intensity sweetener(s) as described herein and 2 ppm to 10 ppm of one or more low-potency sweetener(s) as described herein.

[0059] In some embodiments, an edible composition comprises at least one sweetener and a sweet taste modifying composition consisting of 20 ppm to about 30 ppm of one or more high-intensity sweetener(s) as described herein and 2 ppm to 10 ppm of one or more low-potency sweetener(s) as described herein. In some embodiments, an edible composition comprises at least one sweetener and a sweet taste modifying composition consisting of 22 ppm to about 28 ppm of one or more high-intensity sweetener(s) as described herein and 2 ppm to 5 ppm of one or more low-potency sweetener(s) as described herein. In some embodiments, the high-intensity sweetener is mogroside V. In some embodiments, the low-potency sweetener is 11-O-mogroside V.

[0060] One or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can be present in the sweetened composition in a total amount equal to or greater than about 15 ppm. For example, one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can be present in the sweetened composition in a total amount equal to or greater than about 16 ppm, or equal to or greater than about 17 ppm, or equal to or greater than about 18 ppm, or equal to or greater than about 19 ppm, or equal to or greater than about 20 ppm, or equal to or greater than about 21 ppm, or equal to or greater than about 22 ppm, or equal to or greater than about 23 ppm, or equal to or greater than about 24 ppm, or equal to or greater than about 25 ppm.

[0061] For example, one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can be present in the sweetened composition in a total amount of less than or equal to about 50 ppm, such as less than or equal to about 45 ppm, such as less than or equal to about 40 ppm, or less than or equal to about 35 ppm. For example, one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can be present in the sweetened composition in a total amount ranging from about 15 ppm to about 50 ppm, or from about 15 ppm to about 45 ppm, or from about 15 ppm to about 40 ppm, or from about 15 ppm to about 35 ppm, or from about 20 ppm to about 35 ppm, or from about 20 ppm to about 30 ppm.

[0062] The term "ppm" refers to one or more parts per million by weight, e.g., the weight (in milligrams) of a compound such as mogroside V per kilogram of product containing such compound (i.e., mg / Kg), or the weight (in milligrams) of a compound such as mogroside V (e.g., an oral consumable / edible product of the present disclosure) per liter of product containing such compound (i.e., mg / L), or to one or more parts per million by volume, e.g., the volume (in milliliters) of a compound such as mogroside V per liter of product containing such compound (i.e., ml / L).

[0063] The sweet taste altering compositions described herein may, for example, contain higher concentrations of high-intensity and low-intensity sweeteners and then be diluted in sweetened compositions to obtain the concentrations recited herein. A sweetened composition contains at least one sweetener in an amount equal to or greater than its sweetness perception threshold and / or equal to or greater than about 1.5% (w / v) sucrose equivalent. The term "sweetness perception threshold" refers to the lowest known concentration of a composition that can be perceived as sweet by the human palate. A sweetness equal to or greater than about 1.5% (w / v) sucrose equivalent is recognized as "intrinsically sweet" by FEMA.

[0064] At least one sweetener can be nutritive or non-nutritive. Nutritive sweeteners add caloric value to foods that contain them, while non-nutritive sweeteners contain very few calories or no calories at all. Aspartame, the only approved nutritive high-intensity sweetener, contains more than 2% of the calories in an equivalent amount of sugar, as opposed to non-nutritive sweeteners, which contain less than 2% of the calories in an equivalent amount of sugar.

[0065] The at least one sweetener may be selected from, for example, one or more of sucrose, fructose, glucose, xylose, arabinose, rhamnose, tagatose, allulose, trehalose, isomaltulose, acesulfame potassium (AceK), aspartame, steviol glycoside(s), sucralose, high fructose corn syrup, starch syrup, saccharin, sucralose, neotame, advantame, Monk Fruit Extract, neohesperidin, dihydrochalcone, naringin dihydrochalcone, neohesperidin dihydrochalcone, rubusoside, rebaudioside A, stevioside, stevia, trilobutein, and sugar alcohols such as erythritol, xylitol, mannitol, sorbitol, and inositol. Examples of sweeteners that can be used in the sweetened compositions are disclosed, for example, in WO 2016 / 038617, the contents of which are incorporated herein by reference.

[0066] The at least one sweetener may be selected from, for example, one or more of sucrose, high fructose corn syrup, acesulfame potassium (AceK), aspartame, steviol glycoside(s), and / or sucralose. How to sweeten consumables using a sufficient amount of sweetener is well known in the art. Depending on the consumable, the amount of sweetener can be reduced by adding the sweetness modifying composition as described herein. For example, a reduction of Brix of about 1° to about 4° or more can be achieved.

[0067] At least one other sweetener present in an amount equal to or greater than its sweetness perception threshold and / or in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent may be used in the sweetened composition in an amount equal to or greater than about 0.01% (w / v). For example, at least one other sweetener may be used in the sweetened composition in an amount equal to or greater than about 0.1% (w / v), or in an amount equal to or greater than about 0.5% (w / v), or in an amount equal to or greater than about 1% (w / v), or in an amount equal to or greater than about 2% (w / v). For example, at least one other sweetener may be used in the edible composition in an amount equal to or less than about 20% (w / v), or in an amount equal to or less than about 15% (w / v), or in an amount equal to or less than about 10% (w / v), or in an amount equal to or less than about 8% (w / v), or in an amount equal to or less than about 6% (w / v), or in an amount equal to or less than about 5% (w / v). At least one other sweetener present in an amount equal to or greater than its sweetness perception threshold and / or in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent may be used in the sweetened compositions (e.g., edible compositions) disclosed herein in an amount isosweet to about 2% (w / v) to about 15% (w / v) sucrose.

[0068] In some embodiments, provided herein are sweet taste modifying compositions consisting of mogroside V and 11-O-mogroside V in a ratio ranging from about 2:1 to about 12:1, e.g., from about 6:1 to about 10:1. The sweet taste modifying composition can be used as a sweetness enhancer or modifier in an edible composition. The edible composition can contain at least one other sweetener, such as sucrose. Mogroside V can be used in an edible composition in an amount ranging from about 15 ppm to about 30 ppm, or from about 20 ppm to about 30 ppm (e.g., about 20 ppm or about 25 ppm). 11-O-mogroside V can be used in an edible composition in an amount ranging from about 2 ppm to about 12 ppm, or from about 2 ppm to about 10 ppm (e.g., about 8.5 ppm or about 3 ppm). At least one other sweetener is present in the edible composition, for example, in an amount isosweet to about 2% (w / v) to about 15% (w / v) sucrose.

[0069] The composition may be in any suitable form, for example, a solid (e.g., powder, granules, tablet) or a solution (e.g., an aqueous solution), or an emulsion, or a suspension. For example, the composition may further comprise a diluent or a bulking agent such as dietary fiber. Edible compositions as disclosed herein include, for example:

[0070] - Wet / liquid soups of any consistency or container, including frozen soups. For purposes of this definition, soup(s) means food prepared from meat, poultry, fish, vegetables, grains, fruits, and other ingredients, cooked in a liquid that may contain visible particles of some or all of these ingredients. It may be clear (as a broth) or thick (as a chowder), smooth, strained, or filled, instant, semi-concentrated, or concentrated, and may be served hot or cold, as a first or main course of a meal, or as a snack (sipped like a beverage). Soups may be used as an ingredient for preparing other meal components and may range from broths (consomme) to sauces (cream- or cheese-based soups). - Dehydrated cooking foods, including cooking aid products such as powders, granules, pastes and concentrated liquid products, including concentrated bouillons, bouillons and bouillon-like products in pressed cubes, tablets or powder or granular form, sold separately as finished products or as ingredients in products, sauces or recipe mixes (regardless of technology).

[0071] - Meal solution products, such as: dehydrated or ambient preparations of dehydrated soup mixes, dehydrated instant soups, dehydrated freeze-dried soups, including dehydrated ready-to-cook soups, ready-to-eat meals, meals and single-serve entrees, including pasta, potato and rice dishes - Meal decoration products, such as: condiments, marinades, salad dressings, salad toppings, dips, batters, batter mixes, shelf-stable spreads, barbecue sauces, liquid recipe mixes, concentrates, sauces or sauce mixes (including recipe mixes for salads, whether dehydrated, liquid or frozen, sold as a finished product or as an ingredient within a product).

[0072] - Beverages, including beverage mixes and concentrates, including but not limited to alcoholic and non-alcoholic instant dry powdered beverages, carbonated and non-carbonated beverages (e.g., soda, fruit or vegetable juices, alcoholic and non-alcoholic beverages). Confectionery products, such as cakes, cookies, pies, candy, chewing gum, gelatin, ice cream, sorbet, pudding, jams, jellies, salad dressings and other condiments, cereals and other breakfast foods, canned fruit and fruit sauces, etc. - Dairy products such as milk, cheese, and yogurt.

[0073] - Pharmaceutical compositions, which may be, for example, in the form of a syrup, emulsion, suspension, solution or other liquid form. - Dental compositions including, for example, mouth fresheners, mouthwashes, mouth rinses, toothpastes, tooth polishes, dentifrices, mouth sprays and dental floss. - Edible gel composition.

[0074] The compositions disclosed herein may further comprise a base composition. For example, the edible compositions disclosed herein may further comprise an edible base composition. This refers to all the components required for the composition, except for the combination of a high-intensity sweetener and a low-potency sweetener (e.g., a sweetness-modifying composition). The base composition may be, for example, a sweetened base composition containing at least one other sweetener present in an amount equal to or above its sweetness perception threshold and / or in an amount equal to or above about 1.5% (w / v) sucrose equivalent. Naturally, both the nature and proportions of these may vary depending on the nature and use of the composition, but they are well known in the art and can be used in art-recognized proportions. The formulation of such base compositions for any conceivable purpose is therefore within the ordinary skill in the art.

[0075] Ingredients in the base composition may include, but are not limited to, anti-caking agents, anti-foaming agents, antioxidants, binders, colorants, diluents, disintegrants, emulsifiers, encapsulating agents or formulations, enzymes, fats, flavor enhancers, flavoring agents, gums, lubricants, polysaccharides, preservatives, proteins, solubilizers, solvents, stabilizers, sugar derivatives, surfactants, sweeteners, vitamins, waxes, etc. Solvents that can be used are known to those skilled in the art and include, for example, ethanol, ethylene glycol, propylene glycol, glycerin, and triacetin. Encapsulating agents and gums include maltodextrin, gum arabic, alginates, gelatin, modified starch, and polysaccharides.

[0076] Examples of additives, excipients, carriers, diluents or solvents for flavor or fragrance compositions can be found, for example, in "Perfume and Flavor Materials of Natural Origin", S. Arctander, Ed., Elizabeth, NJ, 1960; "Perfume and Flavor Chemicals", S. Arctander, Ed., Vol. I & II, Allured Publishing Corporation, Carol Stream, USA, 1994; "Flavorings", E. Ziegler and H. Ziegler (ed.), Wiley-VCH Weinheim, 1998, and "CTFA Cosmetic Ingredient Handbook", J.M. Nikitakis (ed.), 1st ed., The Cosmetic, Toiletry and Fragrance Association, Inc., Washington, 1988.

[0077] The ratio of one or more high-intensity sweetener(s) to one or more low-potency sweetener(s) in combination (e.g., sweet taste modifying composition) or one or more sweetness enhancers selected from mogroside IV, siamenoside, and neomogroside depends on the nature of the composition and the desired sweetness intensity and sweetness characteristics. Those skilled in the art can easily ascertain the appropriate ratio in any case by simple, non-inventive experimentation. The amounts and ratios disclosed herein are merely exemplary, and flavorists may seek specific effects by working outside these ranges, and should be considered merely indicative.

[0078] The pH of the compositions disclosed herein may be any pH that does not adversely affect the taste of the sweetener blend. For example, the pH may range from about 1.8 to about 8, or from about 2 to about 5. Those skilled in the art will be able to identify the appropriate concentration of each sweetener used depending on the pH of the composition.

[0079] The use of one or more low-potency sweetener(s) with one or more high-intensity sweetener(s) may improve one or more sweetness characteristics in the sweetened composition, for example, compared to the use of one or more high-intensity sweetener(s) alone. Thus, the sweetened compositions disclosed herein may have one or more improved sweetness characteristics, for example, compared to a sweetened composition in the absence of one or more low-potency sweetener(s). The use of one or more sweetness enhancers selected from mogroside IV, siamenoside, and neomogroside may improve one or more sweetness characteristics of the sweetened composition, for example, compared to the use of a different sweetness enhancer, such as swingle extract, in place of one or more of mogroside IV, siamenoside, and neomogroside.

[0080] The sweetened compositions disclosed herein may have one or more sweetness characteristics that are more similar to sucrose, for example, compared to the sweetened composition in the absence of the one or more low-potency sweetener(s) or compared to a sweetened composition that includes a different sweetness enhancer.

[0081] The sweetened compositions disclosed herein may have, for example, a reduced, lingering sweet taste compared to the sweetened composition in the absence of one or more low-potency sweetener(s) or compared to a sweetened composition containing a different sweetness enhancer.

[0082] The sweetened compositions disclosed herein may have, for example, a reduced bitter and / or astringent and / or metallic and / or licorice-like taste compared to the sweetened composition in the absence of one or more low-potency sweetener(s) or compared to a sweetened composition containing a different sweetness enhancer.

[0083] The sweetened compositions disclosed herein may have an enhanced sweetness impact, for example, compared to a sweetened composition in the absence of one or more low-potency sweetener(s) or compared to a sweetened composition that includes a different sweetness enhancer.

[0084] The comparative sweetened composition is identical except that it does not include any of the one or more low-potency sweetener(s), or it is identical except that it contains a different sweetness enhancer in place of one or more mogroside(s) (e.g., one or more of mogroside IV, siamenoside, and neomogroside).

[0085] use Provided herein is the use of a combination of one or more high-intensity sweetener(s) and one or more low-potency sweetener(s) to enhance the sweetness of a composition containing at least one other sweetener present in an amount equal to or above its sweetness perception threshold and / or in an amount equal to or above about 1.5% (w / v) sucrose equivalent. The combination of one or more high-intensity sweetener(s) and one or more low-intensity sweetener(s) has a sweetness of less than 1.5% (w / v) sucrose equivalent. The one or more high-intensity sweetener(s), one or more low-potency sweetener(s), and at least one other sweetener may be according to any embodiment disclosed herein.

[0086] Provided herein is the use of one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, to enhance the sweetness of a composition comprising at least one other sweetener present in an amount equal to or above its sweetness perception threshold and / or in an amount equal to or above about 1.5% (w / v) sucrose equivalent.

[0087] Thus, there is provided a method for enhancing the sweetness of a sweetened composition, comprising providing a base composition having a sweetness above its sweetness perception threshold and / or containing at least one sweetener in an amount equal to or greater than about 1.5% (w / v) sucrose equivalent, and adding at least one low-potency sweetener, at least one high-intensity sweetener, or one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside. The components of the final composition may be added in any order to obtain the desired final composition. The method may, for example, include mixing the components.

[0088] One or more high-intensity sweeteners, and / or a combination of one or more high-intensity sweeteners with one or more low-potency sweeteners (e.g., a sweet taste modifying composition), and / or one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can, for example, increase the sweetness of a sweetened composition by about 1.0% (w / v) sucrose equivalent or more. For example, high-intensity sweetener(s), and / or a combination of high-intensity sweetener(s) with low-potency sweetener(s), and / or one or more mogroside(s), such as one or more of mogroside IV, siamenoside, and neomogroside, can increase the sweetness of the sweetened composition by about 1.1% (w / v) sucrose equivalent or more, or about 1.15% (w / v) sucrose equivalent or more, or about 1.2% (w / v) sucrose equivalent or more, or about 1.25% (w / v) sucrose equivalent or more. The composition can, for example, contain at least one other sweetener.

[0089] Also provided herein is the use of one or more low-potency sweetener(s) to improve one or more sweetness characteristics of a sweetened composition containing one or more high-intensity sweetener(s), wherein the one or more high-intensity sweetener(s) and the one or more low-potency sweetener(s) are used in an amount having a sweetness of less than about 1.5% (w / v) sucrose equivalent.

[0090] Thus, there is provided a method for improving one or more sweetness characteristics of a sweetened composition comprising one or more high-intensity sweetener(s) in an amount having a sweetness of less than 1.5% (w / v) sucrose equivalent, the method comprising providing one or more high-intensity sweetener(s) and adding one or more low-potency sweetener(s). The components of the final composition may be added in any order to obtain the desired final composition. The method may, for example, include blending the components.

[0091] Improving one or more sweetness characteristics of a sweetened composition that includes a high intensity sweetener can, for example, provide a sweetness characteristic that is more similar to the sweetness characteristic of sucrose.

[0092] Sweetness characteristics may also refer to flavor profile, which refers to the intensity of flavor and sensory attributes of a given compound. Exemplary flavor attributes of sweetness are sweet intensity, bitterness, dark licorice, etc.

[0093] Sweetness profile may refer to a temporal profile, which refers to the change in sweetness perception over time. Every sweetener exhibits a characteristic onset time (AT) and extinction time (ET). Most high-potency sweeteners, in contrast to carbohydrate sweeteners, exhibit a prolonged ET (lingering). Generally, the detected sucrose equivalent spikes to a maximum response level and then tapers off over time. The longer the taper, the greater the lingering sweetness detected from the compound.

[0094] In some embodiments, one or more low-potency sweetener(s) can be used to attenuate the lingering sweet taste of a sweetened composition containing one or more high-intensity sweetener(s). In other words, a low-potency sweetener can be used to reduce the extinction time (ET) of a sweetened composition containing one or more high-intensity sweetener(s). This relates to the undesirable lingering sweet taste in the mouth after the composition is initially ingested or exhaled. Lingering sweet taste refers, for example, to the length of time the sweet taste lingers after initial detection, how quickly the intensity of the sweet taste diminishes or fades after initial detection, and the intensity of the sweet taste after initial detection. One or more low-potency sweetener(s), for example, can decrease the length of time the sweet taste lingers after initial detection and / or increase the speed at which the sweet taste diminishes after initial detection and / or decrease the intensity of the sweet taste after initial detection.

[0095] In some embodiments, one or more low-potency sweetener(s) may be used to counteract the bitter and / or astringent and / or metallic and / or licorice-like taste of a sweetened composition containing one or more high-intensity sweetener(s). The term "licorice-like" refers to the sweet taste of a compound. In some embodiments, one or more low-potency sweetener(s) can be used to enhance the sweetness impact of a sweetened composition containing one or more high-intensity sweetener(s). Sweetness impact is related to the length of time before sweetness is first detected and the intensity of sweetness when it is first detected. One or more low-potency sweetener(s), for example, can decrease the amount of time before sweetness is first detected and / or increase the intensity of sweetness when it is first detected. Sweetness intensity and other sweetness attributes described herein can be evaluated by a trained expert tasting panel, for example, as described in the examples below.

[0096] Manufacturing method Further provided herein are methods of making the compositions disclosed herein. The compositions may be according to any of the embodiments disclosed herein. The method may include combining the components of the desired composition in the desired proportions and, optionally, blending the components. The components may be combined and blended in any suitable order.

[0097] Those skilled in the art will be able to identify suitable methods for making the compositions (e.g., suitable orders for combining or mixing the components) depending on the nature of the composition and the level and characteristics of sweetness desired. The methods may include, for example, providing a desired base composition and adding a sweetener thereto.

[0098] Each of the sweeteners disclosed herein may be made by synthetic processes, or by biological (e.g., enzymatic) or fermentation processes, or may be isolated from natural sources such as plants or fruits. The process may, for example, include contacting at least one mogrol precursor substrate with a mogrol pathway enzyme. The enzyme may, for example, be present in a cell lysate or present in a host cell (e.g., a recombinant host cell). The enzyme may, for example, be a UGT enzyme (UDP-glucuronosyltransferase). For example, mogroside compounds may be made by the biosynthetic pathways disclosed in WO 2013 / 076577 or WO 2014 / 086842, the contents of which are incorporated herein by reference.

[0099] For example, mogroside V may be made by the biosynthetic pathway disclosed in Itkin et al., "The biosynthetic pathway of the nonsugar, high-intensity sweetener mogroside V from Siraitia grosvenorii," PNAS, November 7, 2016, E7619-E7628 and WO 2016 / 038617, the contents of which are incorporated herein by reference. For example, a mogroside compound may be made by modifying (e.g., redistributing) the glycoside content of another mogroside compound. For example, a mogroside compound may be made by redistributing the glycoside content of another mogroside compound using an acid or enzyme as disclosed in WO 2014 / 150127, the contents of which are incorporated herein by reference.

[0100] The process may include extracting one or more sweetener compounds from a natural source, such as a plant or fruit. This may be followed by a purification step to produce, for example, a high-intensity sweetener, a low-intensity sweetener, or a mixture of sweeteners (e.g., a mixture of high-intensity sweeteners, such as a mixture of mogrosides). The extract may have, for example, a relatively high content of mogroside V and / or 11-O-mogroside V (e.g., at least about 30 wt% or at least about 40 wt%). This may involve, for example, fractionation, such as by flash column chromatography. One or more mogroside compounds (e.g., mogroside V) may be obtained from the fruit of Siraitia grosvenorii.

[0101] When a fermentation process is used to produce a target compound (e.g., a target mogroside compound), the target can be extracted from the aqueous fermentation reaction medium using a suitable solvent (e.g., heptane), followed by fractional distillation. The chemical composition of each fraction can be quantitatively determined by GC / MS (gas chromatography-mass spectrometry). The fractions can be blended to produce the desired mogroside compound (e.g., mogroside V and 11-O-mogroside V) for use in flavors or other applications. Verification of the acceptability of the final blended product can be performed by direct comparison with a reference mogroside flavoring product (eg, an existing natural flavoring product obtained from a commercial source).

[0102] example Example 1 method Luo Han Guo fruit extracts obtained from Blue California (Tomas, Rancho Santa Margarita, California) (Extract 4), Azile LCC (Rolling Hills Est, California) (Extracts 1 and 2), and Chr. Olesen Group (Gentofte, Denmark) (Extract 3) were analyzed to identify compounds present in the extracts. A sample solution of the extract was prepared by dissolving 16.52 μg of sample in 25.0 mL of solvent (acetonitrile / water 20 / 80 v / v). From this solution, 100 μL was transferred into an HPLC vial and 900 μL of solvent was added (66.1 ppm solution). From the sample solution, 10 μL was transferred into an HPLC vial and 990 μL of solvent was added (6.61 ppm solution). Both the 66.1 and 6.61 ppm solutions were injected twice onto the LC-MS.

[0103] A calibration (standard) solution of mogroside V was made by dissolving 9.22 mg of mogroside V (98.5% mogroside V obtained from AAPIN chemicals Ltd., Oxfordshire, UK) in 10.0 mL of solvent (acetonitrile / water 20 / 80 v / v). The stock solution was kept in a refrigerator and used to prepare solutions of mogroside V at various concentrations (0.11 ppm, 0.34 ppm, 1.02 ppm, 3.07 ppm, and 9.22 ppm). These solutions were also injected twice onto the LC-MS.

[0104] 2 μL of each solution was injected onto an Acquity C18 BEH 1.7 μm 150 × 2.1 mm column (Waters, Milford, Massachusetts, United States) at 40 °C. Compounds were eluted using a mixture of acetonitrile and 0.1% formic acid in water, starting from 20% acetonitrile and increasing to 50% acetonitrile over 14 min. The gradient was returned to the initial value over 1 min and allowed to stabilize for 5 min. The flow rate was set to 400 μL throughout the run. Eluted compounds were detected using liquid chromatography-mass spectrometry (LC / MS). The mass spectrometer was operated in ESI negative mode, measuring 150-2000 Amu at a resolution of 70,000. The gas flow rates were 60 sheath, 20 auxiliary, and 3 sweep. The capillary and auxiliary gas heater temperatures were set at 380°C and 400°C, respectively.

[0105] The % of each component in the solution was calculated using the following equation and calibrated against the curve of various concentrations of the calibration mogroside V (standard) solution described above.

number

[0106] result FIG. 1 shows the chromatogram of the Monk Fruit extract (Extract 2 in Table 1 below). Figure 1 shows the composition of four different Monk Fruit extracts. Mogroside V is the mogroside with the highest concentration in all four extracts (approximately 45 wt% in Extract 1).

[0107] [Table 1-1] [Table 1-2] [Table 1-3]

[0108] Example 2 method A Monk Fruit Extract (Extract 1 in Example 1 above) obtained from Azile LCC (Rolling Hills Est, California), containing approximately 68 wt% mogrosides, was fractionated by reverse-phase (C-18) flash chromatography. Compounds were eluted using a mixture of methanol (MeOH) in water, starting with 30% MeOH, followed by a linear gradient from 30 to 80% MeOH, and then finally flushing the column with 80% MeOH. The solvent was introduced at a flow rate of 30 ml / min throughout the separation procedure. Eluted compounds were visualized with a UV detector set at 210 nm and a corona light scattering detector. The % of each component in the extract was calculated using the equations described in Example 1 above.

[0109] The collected fractions were pooled according to Table 2 below and freeze-dried into powder. The powders corresponding to the various pooled fractions as given in Table 2 below were layered on top of 5% sucrose and dissolved at various concentrations. The taste of these samples was compared with the 5% sucrose control by three expert panelists (trained flavorists). This determined the sweetness-enhancing effect of each fraction or pool of fractions observed in 5% sucrose.

[0110] result The results are shown in the table below. The total extract was collected in 22 fractions. Fractions 1-10 do not contain mogroside V.

[0111] [Table 2-1] [Table 2-2]

[0112] Fractions 1-10 and 19-22 have the off-taste characteristic of Luo Han Guo fruit and do not have the effect of enhancing sweetness. A better sweetness-enhancing effect was observed in fractions 12-17, which contain mainly mogrosides. When fractions contain almost pure mogroside V, such as fractions 14 and 15, the lingering, harsh, fermented notes are more prominent. Thus, pure mogroside V has the inherent lingering off-taste.

[0113] Fraction 12 had the cleanest sweetness but was less upfront due to a small percentage of mogroside V. Fraction 13 had better sweetness quality but slightly more astringency. 11-O-mogroside V and mogroside V are the two major mogrosides in these two fractions, but in different ratios (F12 mogroside V:11-O-mogroside V is 4:9, and F13 mogroside V:11-O-mogroside V is 13:3). Mogroside V was judged to be extremely sweet, 425 times sweeter than sucrose, while 11-O-mogroside V was considered 84 times sweeter than sucrose.

[0114] Example 3 method A Monk Fruit Extract (Extract 1 in Example 1 above) obtained from Azile LCC (Rolling Hills Est, California), containing approximately 68 wt% mogrosides, was fractionated, and the composition of each fraction was determined by the chromatographic method described above in connection with Example 1. Each fraction was combined with a solution of 5% sucrose and the taste of these samples was compared to a 5% sucrose control by three expert panelists (trained flavorists).

[0115] result Figure 3 shows the chemical composition of fractions 11 to 20 of the extract.

[0116] [Table 3-1] [Table 3-2] [Table 3-3]

[0117] Table 4 shows the tasting results for fractions 12 to 15. The results are similar to those obtained in Example 2.

[0118] [Table 4]

[0119] Example 4 Mogroside V and 11-O-mogroside V were isolated from the Momordica extract using an Agilent 1100 preparative HPLC system with a Phenomenex Luna C18 (2) column (5 μm, 210 × 21.4 mm) and combined to form solutions of various concentrations. These solutions were combined with a solution containing 5% (w / v) sucrose and 0.03% (w / v) citric acid to give test samples, which were evaluated by five experts (trained flavorists) of a sweet-sensitive taste panel. The results are shown in Table 5 below.

[0120] [Table 5]

[0121] Surprisingly, it was found that blending 11-O-mogroside V with mogroside V improved the sweetness quality compared to mogroside V alone. Layering 11-O-mogroside V with mogroside V helped reduce the sweetness linger (diluting the later sweetness) and the astringent, bitter aftertaste compared to mogroside V alone. Thus, 11-O-mogroside V made the sweetness taste more like sugar (i.e., helped provide a temporal profile closer to that of sugar) than mogroside V alone. This allowed for the use of higher concentrations of mogroside V to achieve a higher degree of sweetness while eliminating the disadvantages associated with using this sweetener at higher concentrations (e.g., lingering, bitter, and astringent aftertaste). This was surprising considering that mogroside V is the most potent mogroside sweetener and 11-O-mogroside V is significantly less potent.

[0122] Example 5 A sweet-sensitive taste panel ranked the sweetness of a solution of a sweetener blend ("Mixture 1") relative to sucrose solutions at a range of concentrations to determine sucrose equivalents. The blend was a combination of fractions 11-18 from Example 2 and contained 8.16 wt% 11-O-mogroside V and 61.6 wt% mogroside V. The results are shown in Table 6 below.

[0123] [Table 6]

[0124] The data demonstrate that Mix 1 has a sweetness (as determined by seven panelists) of less than 1% sucrose equivalent, which is recognized by FEMA as "essentially non-sweet." Therefore, Mix 1 is suitable for use as a sweetness modifier or blend at the concentrations noted, since it has no detectable sweetness at these levels.

[0125] A concentration of Mixture 1 with a sweetness threshold close to 1% (35 ppm) was selected and added to a 5% (w / v) sucrose solution. This solution was then ranked against 5, 6, 6.5, and 7% (w / v) sucrose solutions. This was repeated for the 45 ppm Monk Fruit extract. The average score for each solution was determined. The results are shown in Table 7 below.

[0126] [Table 7]

[0127] It was surprisingly found that Blend 1 and Monk Fruit extract acted as sweetness enhancers, as the increase in sweetness in a 5% (w / v) sucrose solution to which they were added exceeded the sweetness of the sweeteners alone.

[0128] The taste of various concentrations of Mixture 1 was tested by a panel of three expert flavorists in a solution containing 5% sucrose and 0.03% citric acid. The taste was compared to the Monk Fruit extract used in Example 2 (obtained from Azile LCC, Rolling Hills Est., California (Extract 1 in Example 1 above) and containing approximately 68 wt.% mogrosides) in combination with the same 5% sucrose and 0.03% citric acid solution. The results are shown in Table 8 below.

[0129] [Table 8]

[0130] Overall, Blend 1 provides better sweetness quality (less harsh, sweeter) than the swingle extract.

[0131] Example 6 A sweet-sensitive taste panel ranked the sweetness of solutions of various sweeteners (mogroside V, mogroside IV, siamenoside, neomogroside, and 11-O-mogroside V) relative to sucrose solutions at a range of concentrations to determine sucrose equivalents. Sweeteners were obtained using an Agilent 1100 preparative HPLC system with a Phenomenex Luna C18 (2) column (5 μm, 210 × 21.4 mm). Results are shown in Tables 9–13.

[0132] [Table 9]

[0133] [Table 10]

[0134] [Table 11]

[0135] [Table 12]

[0136] [Table 13]

[0137] The data demonstrate that mogroside V (25 ppm), mogroside IV (30 ppm), siamenoside (25 ppm), and the combination of mogroside V (25 ppm) with 11-O-mogroside V (3 ppm) and neomogroside (30 ppm) all have sweetness (as determined by seven panelists) below 1.5% sucrose equivalent, which is recognized by FEMA as "essentially non-sweet." Thus, these compounds and mixtures are suitable for use as sweetness modifiers or blends at the concentrations noted, since they have no detectable sweetness at these levels.

[0138] The sweetener concentrations to be tested were selected with an isosweetness threshold close to 1% and added to a 5% (w / v) sucrose solution. These solutions were then ranked against 5, 6, 6.5, and 7% (w / v) sucrose solutions. The average score for each solution was determined. The results are shown in Table 14 below.

[0139] [Table 14]

[0140] It was surprisingly found that mogroside V, siamenoside, neomogroside and mogroside V act as sweetness enhancers, since the increase in sweetness in a 5% (w / v) sucrose solution to which they were added exceeded the sweetness of the sweeteners alone.

[0141] The taste of these sweeteners was tested by a three-person expert panel (trained flavorists) in a solution containing 5% sucrose and 0.03% citric acid. The taste was compared with the Monk Fruit extract used in Example 2 (obtained from Azile LCC, Rolling Hills Est., California (Extract 1 in Example 1 above) and containing approximately 68 wt.% mogrosides). Mogroside IV, siamenoside, and neomogroside all outperformed the Monk Fruit extract in terms of sweetness quality when added to 5% sucrose and 0.03% citric acid. These three compounds provide a sugar-like taste with less lingering sweetness. Siamenoside was described as having "a sweeter body, a sweeter, rounder, slightly more upfront, and a rounder, more persistent sweetness." Mogroside IV was described as having "good sweetness, similar to that of mogroside V." Neomogroside was described as having a "sweet, but slightly bitter aftertaste." The results for mogroside V are shown in Table 15.

[0142] [Table 15]

[0143] In general, tasters agreed that 80% mogroside V did not have the same fully rounded sweetness profile as the swingle extract. 80% mogroside V was more acidic tasting when applied to a sugar / acid / water solution.

[0144] Example 7 method Mogroside V, siamenoside, mogroside IV, and neomogroside were obtained using an Agilent 1100 preparative HPLC system with a Phenomenex Luna C18 (2) column (5 μm, 210 × 21.4 mm). Mogroside V, siamenoside, mogroside IV, and neomogroside were each added to a solution containing 5% sucrose and 0.03% citric acid at concentrations of 25 ppm (mogroside V), 25 ppm (siamenoside), 30 ppm (mogroside IV), and 30 ppm (neomogroside), respectively.

[0145] These test solutions were tasted by a panel of seven experts. Each panelist scored the test solution in comparison with the base solution (a solution of 5% sucrose and 0.03% citric acid) for various sweetness aspects (upfront sweetness, overall sweetness, lingering sweetness, astringency, and volatile off-taste). A score of 0 indicated the same taste aspect, 1 indicated slightly higher, 2 indicated higher, 3 indicated significantly higher, -1 indicated slightly lower, -2 indicated lower, and -3 indicated significantly lower. The average score of each test solution for each taste aspect was calculated. The results are shown in Table 16 below.

[0146] [Table 16]

[0147] Example 8 As shown above, siamenoside, neomogroside, and mogroside IV all have similar or better sweet taste qualities layered on 5% sucrose and 0.03% citric acid compared to mogroside V. Therefore, the taste of 11-O-mogroside V plus each of these mogrosides is evaluated as shown in Table 17.

[0148] [Table 17]

[0149] Example 9 The taste of mogroside V with and without 11-O-mogroside V in various milk or yogurt bases is evaluated as shown in Table 18. The isosweetness threshold for mogroside V in milk and yogurt is also evaluated. The milk base (2% fat) includes 2% fat milk and 5% sucrose. The nonfat yogurt base includes plain nonfat yogurt and 5% sucrose. The full-fat yogurt base includes plain full-fat yogurt and 5% sucrose. Higher dosage levels are used for milk and yogurt due to the fat, protein, and other components. Monk fruit extract is used at 75 ppm for these applications.

[0150] [Table 18]

[0151] Example 10 Identification of a novel minor cucurbitane glycoside from Siraitia grosvenorii Introduction Siraitia grosvenorii (Swingle) C. Jeffrey ex Lu et ZY Zhang is a perennial vine in the Cucurbitaceae family, endemic to southern China and northern Thailand. The fruit of S. grosvenorii, commonly known as "monk fruit," has been used in traditional Chinese medicine for hundreds of years to treat respiratory infections, bronchitis, gastritis, constipation, and other conditions. Recent pharmacological studies have confirmed that S. grosvenorii extracts or their constituents possess various biological activities, including antibacterial, anti-inflammatory, antidiabetic, anticancer, and immunostimulatory properties. [1] Monk fruit is used as a sweetener in food and beverages in China. It is now one of the most well-known natural high-intensity sweeteners worldwide. Since the discovery of the cucurbitane glycoside mogroside V as the sweetener of S. grosvenorii, more than 40 cucurbitane triterpenoids have been reported from S. grosvenorii so far [1-4].

[0152] Researchers in the food and flavor industries are actively discovering and adding more novel compounds to the mogroside pool in an effort to find novel mogrosides with better sweetening properties [5-7]. Novel molecules with better sweetening properties under the category of known natural sweeteners have been highly sought after by the food and flavor industries. The commercialization of rebaudioside M (also known as rebaudioside X) is a good example. Despite being a minor natural product (less than 0.1%) from stevia discovered in 2010, rebaudioside M quickly progressed to the commercialization stage, benefiting from cost reductions resulting from technological developments in plant biology, molecular biology, and enzyme engineering [8, 9]. Rebaudioside M received Letters of No Objection from the US FDA regarding its Generally Recognized as Safe (GRAS) status in 2013, 2014, and 2017 (GRN numbers 473, 512, and 667) [10-12].

[0153] We have conducted a search to find the best-performing mogrosides or their combinations using commercially available Monk Fruit extracts

[13] . Herein, we report two novel minor cucurbitane glycosides from S. grosvenorii and highlight our novel oligosaccharide elucidation strategy based on HSQC-TOCSY experiments with different mixing times.

[0154] material and method General Experimental Procedures Optical rotations were measured on a Rudolph Autopol IV polarimeter. NMR spectra were recorded on a Bruker DRX Avance 300 or 500 spectrometer. Chemical shifts are given in δ (ppm) relative to the residual solvent peak. Low-pressure chromatography was performed on a Biotage Flash System SP1. Preparative HPLC was performed on an Agilent 1100 preparative HPLC system using a Phenomenex Lunar C18(2) column (5 μm, 210 × 21.4 mm) or a TSKgel Amide-80 (5 μm, 300 × 21.5 mm) (Tosoh Bioscience LLC). Analytical HPLC was performed on an Agilent 1100 analytical HPLC system equipped with a Corona CAD detector. LC-MS was performed using a Waters Q-Tof micro mass spectrometer interfaced with a Waters 2795 separations module.

[0155] plant material Luo Han Guo extract (trade name Swingle, ~60% mogrosides) was purchased from Blue California Co., Ltd. Equipment Chromatography conditions: Chromatography was performed on a Waters Acquity H UPLC. Separation was performed at 25°C using a 1.0 x 100 mm Acquity UPLC HSS T3 column (Waters) with a 1.8 mm particle size and a 0.2 mm prefilter. Solvent A was water, and solvent B was acetonitrile; both solvents contained 0.1% formic acid. The injection volume was set to 10 μl. The chromatographic flow rate was 200 μl / min. The sample was eluted from the LC column using the following linear gradient (curve number 6): 0-40 min: 90% A to 30% A; 40-45 min: 30-10% A; 45-50 min: 10% A; 50-51 min: 10% to 90% A; 51-55 min: 90% A for re-equilibration.

[0156] mass spectrometry The U-HPLC system was coupled to a hybrid quadrupole orthogonal time-of-flight (TOF) mass spectrometer (SYNAPT G2 HDMS, Waters MS Technologies, Manchester, UK) in positive electrospray ionization mode (ESI). + The sampling cone voltage was 40, capillary voltage 0.7 kV, source temperature 40 °C, desolvation temperature 450 °C, desolvation gas flow rate 800 L / h, and cone gas flow rate 50 L / h were optimized. Leucine enkephalin was detected by the lock mass [M+H] at m / z 556.2771. - Sodium formate was used for external instrument calibration.

[0157] purification Three grams of Monk Fruit extract was dissolved in 15 mL of water and loaded onto a pre-equilibrated C-18 Snap cartridge (KP-C18-HS, ​​120 g, 132 mL column volume). The gradient system used (A: water; B: methanol) was: 30% 2 CV, 30%-80% 10 CV, 80%-100% 2 CV, and 100% 2 CV. The flow rate was 30 mL / min. Fractions were collected at 27 mL per tube. A total of 12 g of Monk Fruit was fractionated in four loadings. All fractions were analyzed by analytical HPLC to identify fractions containing the target mogrosides (isocratic mobile phase: 24% acetonitrile in water; column: Luna C18 5 μm 4.6 × 150 mm). Fractions 36–38, carrying iso-mogroside VI and 11-epi-mogroside V, were combined and the solvent was evaporated. Further preparative HPLC purification of fractions 36–38 yielded iso-mogroside VI (1, 22 mg) and 11-epi-mogroside V (2, 17 mg) (24% acetonitrile in water, 10 mL / min, retention times 13.1 and 14.3 min, respectively).

[0158] 11-Oxo-mogroside V (4) and neomogroside (3), with 11-oxo-mogroside V as the major component, were primarily found in flash fractions 39–40. On reversed-phase C-18 preparative HPLC, neomogroside appeared as a tail shoulder of 11-oxo-mogroside (24% acetonitrile in water, 10 mL / min, retention times 17.0 and 18.0 min, respectively). Collection of the peak front afforded 105 mg of compound 11-oxo-mogroside V (4). Further purification of the shoulder neomogroside (3, 15 mg) was achieved by preparative HPLC on TSKgel Amide-80 (65% acetonitrile in water, 20 mL / min, rt 15.5 min).

[0159] Iso-mogroside VI(1): White amorphous powder; [α] 20 D -8.2 (c 0.12,MeOH); 1 H NMR and 13See Table 1 for C spectroscopic data; HRESIMS: m / z 1449.7075 [M+H] - (C 66 H 113 O 34 Calculated value for 1447.7113, Δ2.6 ppm). Epi-mogroside V(2): White amorphous powder; [α] 20 D +4.5 (c 0.13,MeOH); 1 H NMR and 13 See Table 1 for C spectroscopic data; HRESIMS: m / z 1287.6558 [M+H] - (C 60 H 103 O 29 Calculated value for 1287.6585, Δ2.1 ppm).

[0160] Acid hydrolysis of sugars and determination of absolute configuration Compound 1 (1.2 mg) or 2 (1.8 mg) was incubated in 1 mL of 1 M HCl at 80 °C for 3 hours. After hydrolysis, the solution was extracted with EtOAc (1 mL × 3). The remaining aqueous solution was evaporated by blowing nitrogen gas and lyophilized. The absolute configuration of the sugar in the residue was determined by GC-MS analysis of its O-silylated derivative and comparison with the derivatives of D-glucose and L-glucose standards. Briefly, the sugar residue, D-glucose (2 mg) or L-glucose (2 mg), was dissolved in pyridine (0.5 mL). 0.1 M L-cysteine ​​methyl ester hydrochloride (Aldrich, Milwaukee, WI) in pyridine (0.5 mL) was added to the solution. The mixture was kept at 60 °C for 2 hours and dried by blowing nitrogen gas. 1-Trimethylsilylimidazole (Fluka, Buchs, Switzerland) (0.5 mL) was added to the residue and incubated for 1 hour at 60° C. The mixture was separated by adding n-hexane and water (1.0 mL each).

[0161] The n-hexane extract was analyzed by GC-MS under the following conditions: capillary column HP-5MS (30 m × 0.25 mm × 0.25 μm, Agilent); column temperature, 180–230 °C with a gradient of 5 °C / min; injection temperature, 250 °C; carrier, He gas; split ratio, 20:1. O-silylated D-glucose and L-glucose derivatives showed retention times of 16.02 and 16.39 min, respectively. By comparing retention times and cochromatography, the sugar residues after acid hydrolysis of 1 and 2 were determined to be D-glucose.

[0162] Reduction of 11-oxo-mogroside V with NaBH4 25 mg of 11-oxo-mogroside V (4) was dissolved in 50% dioxane, to which 20 mg of NaBH4 was added, and the mixture was heated at 50 °C for 3 days. The reaction mixture was periodically analyzed by HPLC to monitor the progress of the reaction. After the reaction, the mixture was acidified with acetic acid and concentrated to dryness by sparging with nitrogen gas. The residue was redissolved in water and passed through a pre-equilibrated C-18 SPE column. The methanol eluate from the SPE column was concentrated. The residue was then separated by semi-preparative HPLC. The two reduction products had the same retention time and molecular weight as the isolated mogroside V and 11-epi-mogroside V by LS-MS analysis and cochromatography on analytical HPLC. 1-D and 2-D NMR data also confirmed the structures of the two reduction products as mogroside V and 11-epi-mogroside V.

[0163] Results and Discussion Isolation and elucidation of iso-mogroside VI (1) and 11-epi-mogroside (2) During the LC-MS investigation of a commercially available Luo Han Guo extract with 60% mogrosides, several mogrosides with six or five sugar moieties in the extract caught our attention (Figures 2 and 3). Because there have been few reports on the sweetness characteristics of mogroside V and VI isomers, we decided to purify and isolate these isomers for our evaluation. The concentrations of 1, 2, 3, and 4 in the extract were estimated to be 0.8%, 0.5%, 0.6%, and 4.9%, respectively, using a universal Corona detector. After fractionation on a flash chromatography system, followed by preparative HPLC purification, we purified the four targeted mogrosides 1–4 and determined them to be iso-mogroside VI (1), 11-epi-mogroside (2), neomogroside (3), and 11-oxo-mogroside V (4).

[0164] The molecular formula of 1 was determined from its HR-ESI-MS spectral data ([M−H] - m / z, 1447.6957, C 66 H 111 O 34 The calculated value for C is 1447.6957. 66 H 112 O 34 The NMR spectral data of 1 suggested the structure of a hexasaccharide triterpenoid saponin: 30 of the 66 carbons were assigned to the triterpenoid aglycone and 36 of the 66 to the six hexose moieties. 13 C and 1 The H NMR spectrum showed seven singlet tertiary methyl signals, a doublet secondary methyl, and an olefinic methine (Table 19), suggesting the typical (24R)-cucurbit-5-ene-3β,11α,24,25-tetraol mogrol aglycone. 1 H, 13 Further confirmation was obtained by detailed analysis of the C and 2D (COSY, TOCSY, HSQC, and NOESY) NMR data and comparison with the NMR data of mogroside V standards.

[0165] [Table 19-1]

[0166] [Table 19-2]

[0167] GC-MS analysis of the aqueous acid hydrolysis product of 1 showed that D-glucose was the only monosaccharide in the structure of 1. The HSQC spectrum clearly showed six anomeric cross peaks of glucosyls: Glc-I (δ C 106.8 and δ H 4.73), Glc-II(δ C 105.1 and δ H 5.10), Glc-III(δ C 103.7 and δ H 4.85), Glc-IV(δ C 104.6 and δ H 4.78), Glc-V(δ C 104.7 and δ H 5.43), Glc-VI(δ C 104.8 and δ H 5.03). The stereochemistry of all six glucopyranosyls is determined by their anomeric proton coupling constants. 3 J H1, H2 From the results, the β-configuration was determined. From the HSQC-TOCSY experiment (hsqcgpmlph) with a mixing time of 100 ms, the glucopyranosyl carbon signals can be divided into six groups (Figure 4). The oligosaccharide elucidation started from the glucopyranosyl attached to C-3 of the cucurbitan aglycone. Its anomeric proton (δ H 4.73, d, J = 7.9 Hz) and aglycone C-3 (δ C According to the HMBC correlation with 87.6) and the NOESY correlation between Glc-I H-1 and aglycone H-3, Glc-I was determined to be linked to the aglycone C-3. 13 C signal (δ C75.1, 77.8, 71.4, 77.2, 70.1) is δ C The typical C-6 carbon signal around 62 was missing. C The downfield shift of 70.1) indicated glycosylation at this position.

[0168] By comparing the HSQC-TOCSY spectra (hsqcgpmlph) with increasing mixing times from 10, 30, 60, and 100 ms, it can be observed that the magnetization transfer relay gradually spreads from C-2 to C-6 (Figure 4). As shown in Figure 4, HSQC-TOCSY at a mixing time of 10 ms revealed a correlation between H-1 and C-2 of glucopyranosyl. At a mixing time of 30 ms, a correlation between H-1 and C-3 appeared in addition to the correlation between H-1 and C-2. At 60 ms, the carbon chain, as represented by the HSQC-TOCSY correlation, extends to C-4. A complete HSQC-TOCSY correlation between H-1 and C-2 to C-6 can be observed at 100 ms. Therefore, the signals at C-2 to C-6 can be unambiguously assigned. The binding of Glc-II to Glc-I is due to the anomeric Glc-II H-1 (δ H Glc-I C-6 (δ C 70.1) was established by HMBC correlation of Glc-II. 13 C signal (δ C The α-glucan residues (75.0, 77.8, 71.5, 78.0, 62.5) did not suggest glycosylation on Glc-II. Consequently, the sugar chain on the aglycone C-3 was proposed to be 3-O-(β-D-glucopyranosyl(1→6)-β-D-glucopyranosyl).

[0169] Anomeric proton (δ H The aglycone carbon signal (δ 4.85, d, J = 7.5 Hz) C HMBC correlation to 92.3) indicated the attachment of Glc-III H-1 to the aglycone C-24. 13 C pattern (δ CThe C-2 and C-6 downfield shifts (δ 81.6, 78.3, 71.4, 76.4, 70.0) suggested C-2 and C-6 glycosylation shifts. HSQC-TOCSY analysis at mixing times of 10, 30, 60, and 100 ms resulted in sequence assignment and confirmation of the C-2 and C-6 downfield shifts. Glc-IV is attached to C-6 (δ 81.6, 78.3, 71.4, 76.4, 70.0) of Glc-III. C 70.0) and its H-1(δ H 4.78, d, J = 7.5 Hz) HMBC correlation determined that Glc-IV is bound to C-6 of Glc-III. Glc-IV is a normal terminal glucopyranosyl (δ C The linkage of Glc-V to C-2 of Glc-III was anomeric Glc-V H-1 (δ H 5.43, d, J=7.8 Hz) of Glc-III C-2 (δ C 81.6) was established by HMBC correlation to Glc-V H-1 (δ H 5.43) is in agreement with previous reports of similar structures. C C-4 in 70-71 13 C chemical shifts are calculated from the set of Glc-V signals (δ C The amino acid sequence was absent in the 104.7, 75.4, 76.4, 82.0, 76.5, 62.6) suggesting glycosylation at C-4.

[0170] By observing the C-2 to C-6 relay from HSQC-TOCSY at mixing times of 10, 30, 60, and 100 ms, δ C The δ 82.0 was clearly assigned to C-4 of Glc-V (Fig. 4). C 82.0) and H-1 of Glc-VI (δ HThe HMBC crosspeak between Glc-VI and Glc-V (5.03, d, J = 7.7 Hz) further confirmed that Glc-VI was bound to Glc-V at this position. Glc-VI was a terminal glucopyranosyl with no additional sugar chain branches. Based on the above evidence, the structure of iso-mogroside 1 was assigned as 3-O-β-D-glucopyranosyl(1→6)-β-D-glucopyranosyl-mogrol-24-O-β-D-glucopyranosyl-(1→6-[β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl-(1→2)]-β-D-glucopyranosyl.

[0171] Compound 2 was identified based on its HR-ESI-MS data ([M−H] - m / z, 1285.6429), C 60 H 102 O 29 The NMR data of the oligosaccharide portion of 2 were superimposable with those of mogroside V. Detailed 2-D NMR experiments, including HSQC, HMBC, NOESY, COSY, and HSQC-TOCSY, confirmed that 2 had the same sugar moiety as mogroside V. Next, attention was directed to the aglycone NMR data. The HMBC correlation between C-11 and H3-19 confirmed that 2 had the same sugar moiety as mogroside V (δ C 77.8), the C-11 (δ C Further assignment of the aglycone data by 2-D NMR experiments revealed major shifts at C-8, C-10, and C-12 compared with the data for mogroside V. 13 The results showed that C chemical shift changes occurred (Table 19). This suggested a β-OH rather than an α-OH at C-11. The β-OH conformation of 2 was further established by NOE correlations between H-8 and H3-18 and 19; H-10 and H3-28 and H3-30; H-11 and H3-30; and H-17 and H3-30. One natural 11-β-OH cucurbitan and one semisynthetic 11-β-OH cucurbitan have been previously reported [14, 15].

[0172] Compound 2 aglycone 13The C NMR data were in good agreement with those of semisynthetic 11-β-OH cucurbitan glycosides recorded in pyridine-d6

[14] . Matsuda et al. 13 C NMR data were obtained in methanol-d4 and were significantly different in terms of the C-11, C-8, C-10, and C-12 chemical shifts. To further confirm the 11-β-OH structure of 2, semisynthesis of 2 was carried out by chemical reduction of 11-oxo-mogroside V (4) to the 11-β-OH and 11-α-OH isomers of mogroside V. From LC-MS, HPLC cochromatography, and NMR data analysis, semisynthetic 11-epi-mogroside V was determined to be identical to isolated 11-epi-mogroside V. Thus, the structure of 11-epi-mogroside V(2) was elucidated to be 3-O-β-D-glucopyranosyl(1→6)-β-D-glucopyranosyl-11β-OH-mogrol-24-O-β-D-glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranosyl. To our knowledge, this is the first report of a naturally occurring mogroside with an 11β hydroxyl group.

[0173] HSQC-TOCSY at different mixing times for oligosaccharide chain elucidation. Gheysen et al. investigated TOCSY experiments with different mixing times and concluded that 100 ms was the optimal spin-lock time for distinguishing between D-glucose, D-galactose, and D-mannose

[16] . Through their results, we noticed that the spin-lock time could significantly affect the magnetization transfer efficiency of H-1 of D-glucose. The correlation between H-1 and H-2 through H-6 gradually extended to H-6 as the spin-lock time increased. Inspired by their investigation, we hypothesized that by increasing the spin-lock time in HSQC-TOCSY, we should be able to observe that the correlation between H-1 and C-2 through C-6 of glucose gradually extended from C-2 to C-6 as the magnetization transfer chain extended. HSQC-TOCSY with increasing spin-lock time should reveal carbon sequence information, which will be extremely useful for oligosaccharide elucidation and assignment.

[0174] Figure 4 shows the HSQC-TOCSY (hsqcgpmlph) spectra of iso-mogroside VI at mixing times of 10, 30, 60, and 100 ms. The cross-peaks in Figure 4 were quantified by their integrals and compared in Figure 5. Peak intensities (presented as integrals) could, in some cases, be an indication of their distance from H-1. For example, under the 30 ms mixing time experiment, all C-3 peaks were significantly weaker than the C-2 peaks. However, under the 60 ms mixing time, the C-3 peaks became larger than the C-2 peaks. To ensure accurate interpretation, the carbon sequence must be determined not only by the peak intensities under one mixing time but also through an overview of all HSQC-TOCSY spectra under different mixing times.

[0175] Traditionally, NMR elucidation and assignment of saponin glycans starts with the sugar attached to the aglycone. HMBC or NOESY can identify well-resolved anomeric H-1 and C-1 signals. COSY correlation and 3The proton signals of monosaccharides can be assigned through matching of J(H,H) coupling constants. Large coupling constants (>7 Hz) typically represent two adjacent axial C-H bonds, and small coupling constants (<4 Hz) represent axial-equatorial or equatorial-equatorial C-H bonds, allowing the type of monosaccharide to be determined. NOE correlation is useful for confirming the stereochemistry of axial-axial, axial-equatorial, or equatorial-equatorial relationships. The carbon signals (C-2 to C-6) of saccharides are assigned according to HSQC or HMQC. Because the patterns of C-1 to C-6 chemical shifts for different types of monosaccharides are characteristic and consistent, the chemical shifts of the carbon signals as determined by HSQC / HMQC are crucial information for confirming the type of monosaccharide. Through observation of carbon chemical shift changes, glycosylation positions on the glycan can be identified and further confirmed by HMBC correlation. In summary, the traditional approach to elucidating saponin sugars is: HMBC → C-1, H-1 → COSY → H-2~H-6 → HSQC / HMQC → C-2~C-6, then assisted and confirmed by coupling constant analysis and NOESY experiments.

[0176] 1 H- 1 H TOCSY (HOHAHA - Total Correlated Spectroscopy, also known as Homonuclear Hartmann-Hahn) experiments can be of great help in separating the complex sugar proton signals into groups. The transfer of magnetization during TOCSY spin lock from the anomeric H-1 to the end of the furanose or pyranose ring is intermediate. 3The J(H,H) scalar coupling constant depends on the magnitude of the coupling constant. Axial-axial protons with large coupling constants (>7 Hz) allow fast magnetization transfer, whereas axial-equatorial or equatorial-equatorial protons with small coupling constants (<4 Hz) significantly reduce the transfer efficiency. Therefore, TOCSY experiments can not only be used to group proton signals into spin systems, but also provide stereochemical information for saccharides. For example, we should be able to capture the magnetization relay of glucose through H-1 to H-6 with the correct mixing time. For galactose, there is no magnetization relay on H-4, even with a mixing time of 200 ms.

[0177] However, for the case of mogrosides, which have a set of five or six glucopyranosyl signals, using COSY and TOCSY to connect H-1 to H-6 can be quite cumbersome. The proton signals of mogroside glucopyranosyl have very similar chemical shifts, and δ H The proton signals appear clustered in a small range from 3.8 to 4.5. It is difficult to clarify the COSY connections through such poorly resolved proton signals. The glucopyranosyl carbon signals are also very close, and the HSQC cross-peak signals overlap strongly with each other, making resolution and assignment even more difficult. Previously, HSQC-TOCSY has been applied to the structure elucidation and assignment of saponins by grouping together carbon signals in each spin system. [17, 18] Through our investigation, we demonstrated for the first time that the sequence of signals within the glucopyranosyl carbon group can be identified by applying different mixing times in HSQC-TOCSY experiments.

[0178] Figure 6 summarizes the novel HSQC-TOCSY-based strategy for elucidating the glucopyranosyl oligosaccharide chains of mogrosides: In step 1, heteronuclear multibond correlation spectroscopy (HMBC) was used to determine the sugar anomers C-1 and H-1, starting from the sugar linkage to the aglycone. In step 2, HSQC-TOCSY was used with a mixing time of 100 ms to determine the entire group from C-2 to C-6. HSQC-TOCSY or HSQC-TOCSY were used to assign C-2 (d9 = 10 ms), HSQC-TOCSY (d9 = 30 ms) to assign C-3, HSQC-TOCSY (d9 = 60 ms) to assign C-4, and HSQC-TOCSY (d9 = 100 ms) to assign C-5 and C-6. In step 3, if a C-2 downshift from ~δ75 to ~δ81, a C-4 downshift from ~δ71 to ~δ81, or a C-6 downshift from ~δ62 to ~δ69 is observed, check HMBC for glycosylation at these positions. ** If C-2 is downshifted from ~δ75 to ~δ81, C-4 is downshifted from ~δ71 to ~δ81, or C-6 is downshifted from ~δ62 to ~δ69, check HMBC for glycosylation at these positions. **

[0179] 1 H coupling constants and 13 1-D NMR data, such as C carbon signal patterns, and 2-D NMR experiments, such as NOESY, HMBC, TOCSY, COSY, and HSQC, could aid the process and confirm the results. The novel HSQC-TOCSY-based strategy is a simple, rapid, and unambiguous way to elucidate and assign the glucopyranosyl chains of any new or known mogroside. The strategy can also be adapted to elucidate and assign other monosaccharides and oligosaccharides.

[0180] The structures of neomogroside and mogroside VI. Compound 3 was determined to be a neomogroside through detailed 1-D and 2-D NMR analyses and comparison with literature data

[19] . To elucidate the oligosaccharide chain of 3, signals were assigned by HSQC-TOCSY and TOCSY experiments with different mixing times of 10, 30, 60, and 100 ms. The linkages of the six saccharides were determined by their NOESY and HMBC correlations. The oligosaccharide chain on C-3 of the aglycone can be unambiguously assigned as β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl. The glucopyranosyl on C-24 of the aglycone is branched at β-D-glucopyranosyl-(1→2) and β-D-glucopyranosyl-(1→6).

[0181] Neomogroside was first discovered in S. grosvenorii and described by Si et al.

[19] . A search for neomogroside in Scifinder returned the CAS number 189307-15-1. However, an incorrect structure for neomogroside is given in Scifinder, even though the reference cited by Scifinder is the 1996 article by Si et al. The incorrect structure for 189307-15-1 is given in Scifinder as 3-O-β-D-glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→6)]-β-D-glucopyranosyl-mogrol-24-O-β-D-glucopyranosyl-(1→6)-[β-D-glucopyranosyl-(1→2)]-β-D-glucopyranosyl (structure 6 in Figure 2). The neomogroside report by Si et al. was written in Chinese and published in a Chinese journal in 1996. The accessibility and misinterpretation of this article may have led to an incorrect structure in Scifinder.

[0182] In Scifinder, neomogroside and mogroside VI have the same CAS number 189307-15-1 and the same structure. Takemoto et al. first reported mogroside VI from S. grosvenorii [2]. However, it has the molecular formula C 66 H 112 O 34 The article only refers to the pure mogroside, which has the structure ##STR1## and the structure has not been determined. [2] Prakash et al. reported the structure and NMR data of mogroside VI as a known compound in their article published in 2011. [6] In their article, the structure of mogroside VI was assigned as 6 in Figure 2. Prakash stated that the structure of mogroside VI was elucidated by NMR analysis and also by comparison with literature values. However, no citation was given for the literature values.

[0183] For known compounds, comparison of NMR data with literature data could be useful for determining structure. However, the complexity of the NMR data for mogrosides makes it difficult to determine structure primarily by comparison of NMR data with literature data. 1 The 1 H NMR data showed variations in different reports due to different NMR solvents used (the ratio of pyridine to DO could cause signal shifts) or simply incorrect assignments.

[0184] parable 13 The C NMR data are quite consistent, 1 Although the H NMR data have better resolution than the known mogrosides, structural determination of the oligosaccharide chains is difficult. 13 Direct comparison of C NMR data with literature data cannot be relied upon. Considering the case of neomogroside, if Glu-VI glucopyranosyl-(1→2) was branched on Glu-I, Glu-II, Glu-III, Glu-IV, or Glu-V, the five isomers would be very similar. 13 C NMR data can be obtained. Before the oligosaccharide chains of a mogroside can be definitively determined,13 Rather than comparing the C NMR data with literature data, detailed 2-D NMR analysis must be carefully performed.

[0185] Example 11 Sweetness Intensity of Iso-mogroside VI and 11-epi-mogroside V method To make a 100 ppm iso-mogroside VI solution, 10 mg of iso-mogroside VI (Figure 7) was dissolved in 31 mL of water. The working solution of 11-epi-mogroside V (Figure 8) was 374 ppm (9.34 mg of 11-epi-mogroside V in 25 mL of water). A series of standard sucrose solutions was prepared as a sweetness standard (0.50, 0.75, 1.00, 1.25, 1.50%).

[0186] result Four sweet-tolerant panelists were asked to evaluate 100 ppm iso-mogroside VI and 374 ppm 11-epi-mogroside V and a sucrose standard and report their sweetness-equivalent concentrations relative to sucrose. The mean sweetness-equivalent concentrations of each compound were used to calculate isosweetness potency. The isosweetness potency values ​​for iso-mogroside VI and 11-epi-mogroside V were determined to be 91 and 35 times sweeter than sucrose, respectively (100 ppm iso-mogroside VI is equivalent to 0.91% sucrose; 374 ppm 11-epi-mogroside V is equivalent to 1.31% sucrose).

[0187] Example 12 method Iso-mogroside VI and 11-epi-mogroside V were obtained as described in Example 10. Iso-mogroside VI and 11-epi-mogroside V were each added at a concentration of 25 ppm to a solution containing 5% sucrose and 0.03% citric acid. These test solutions were tasted by a panel of seven experts. Each panelist scored the test solution compared to the base solution (5% sucrose and 0.03% citric acid solution) for various aspects of sweetness (overall sweetness, upfront sweetness, lingering sweetness, astringency, and volatile off-notes). A score of 0 indicated the same taste aspect, 1 indicated slightly higher, 2 indicated higher, 3 indicated significantly higher, -1 indicated slightly lower, -2 indicated lower, and -3 indicated significantly lower.

[0188] result The average score for each test solution for each taste dimension was calculated and the results are shown in Table 20 below.

[0189] [Table 20]

[0190] Two novel minor cucurbitane glycosides, along with the known 11-oxo-mogroside and neomogroside, were purified from a commercial extract of swingle fruit (Siraitia grosvenorii (Swingle) C. Jeffrey ex Lu et ZY Zhang). Detailed NMR and LC-MS analyses, as well as chemical synthesis, elucidated the structures of two novel compounds, iso-mogroside VI (1) and 11-epi-mogroside V (2), as follows: 3-O-β-D-glucopyranosyl(1→6)-β-D-glucopyranosyl-mogrol-24-O-(β-D-glucopyranosyl-(1→6)-[β-D-glucopyranosyl-(1→4)-β-D-glucopyranosyl-(1→2)]-β-D-glucopyranosyl and 3-O-β-D-glucopyranosyl(1→6)-β-D-glucopyranosyl-11β-OH-mogrol-24-O-β-D-glucopyranosyl-(1→6)-[β-D-glucopyranosyl-(1→2)]-β-D-glucopyranosyl, respectively.

[0191] The sweetening potencies of iso-mogroside VI and 11-epi-mogroside V were estimated to be 91 and 35 times sweeter than sucrose, respectively (100 ppm iso-mogroside VI is equivalent to 0.91% sucrose; 374 ppm 11-epi-mogroside V is equivalent to 1.31% sucrose). In the course of our identification of novel and known mogrosides bearing five or six glucopyranosyl residues, a new strategy for the elucidation and assignment of glucopyranosyl glycans was developed. The new strategy, based on HSQC-TOCSY with different mixing times, featured rapid and unambiguous elucidation and assignment of glucopyranosyl glycans. After our detailed NMR spectroscopic analysis confirmed the neomogroside structure, previous confusion regarding the structures of neomogroside and mogroside VI was reviewed and clarified.

[0192] The above has broadly described certain aspects of the present invention without limitation. As will be readily apparent to those skilled in the art, variations and modifications are intended to be within the scope of the invention as defined by the appended claims. References

[0193]

Table 21-1

Table 21-2

Claims

1. At least one sweetener present in an amount having a sweetness equivalent to or exceeding approximately 1.5% (w / v) sucrose equivalent; and One or more sweeteners selected from mogroside IV, siamenoside, and neomogroside, having a total sweetness of less than 1.5% (w / v) sucrose equivalent. A sweetened composition containing [the specified ingredient].

2. The sweetened composition according to claim 1, wherein at least one sweetener is selected from the group consisting of sucrose, high-fructose corn syrup, acesulfame potassium (AceK), aspartame, steviol glycoside, and sucralose.

3. The sweetened composition according to claim 1 or 2, wherein the sweetened composition is a moist / liquid soup, a dry and culinary food, a meal solution product, a meal decoration product, a beverage, or a dairy product such as milk, cheese, and yogurt.

4. A sweetened composition according to any one of claims 1 to 3, wherein one or more sweeteners have a purity of at least 80 wt%.

5. The use of one or more of mogroside IV, siamenoside, and neomogroside as sweeteners in a sweetened composition.

6. The use according to claim 5, wherein one or more sweeteners (one or more) are used in a sweetened composition in a total amount having a sweetness of less than 1.5% (w / v) sucrose equivalent.

7. The use according to claim 5 or 6, wherein the sweetened composition is a moist / liquid soup, a dry and cookable food, a meal solution product, a meal decoration product, a beverage, or a dairy product such as milk, cheese, and yogurt.

8. The use according to any one of claims 5 to 7, wherein one or more sweeteners have a purity of at least 80 wt%.

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