AU2011343504A1 - Monatin sweetened food or beverage with improved sweetener performance - Google Patents

Abstract

Monatin compositions including steviol glycosides exhibit an improved sweetener performance, compared to comparable compositions containing monatin in the absence of steviol glycosides. More specifically, after exposure to light, monatin beverage compositions containing rebaudioside A exhibit a reduction in discoloration, a reduction in off-flavors and a higher retention of monatin equivalents, as compared to comparable monatin beverage compositions not containing rebaudioside A. Monatin beverage compositions containing rebaudioside A exhibit decreased sweetness linger as compared to comparable beverage compositions containing monatin without steviol glycosides and exhibit greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation.

Description

WO 2012/083251 PCT/US20111/065643 MONATIN SWEETENED FOOD OR BEVERAGE WITH IMPROVED SWEETENER PERFORMANCE CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims benefit of U.S. Patent Application 61/423,948, filed December 16, 2010, and which is hereby incorporated by reference in its entirety. FIELD [0002] The present disclosure relates generally to a monatin-sweetened food or beverage. Aspects of the disclosure are particularly directed to a inonatin-sweetened food or a beverage containing steviol glycosides, resulting in improved sweetener performance. BACKGROUND [0003] Monatin (2-hydroxy-2-(indol-3-ylmethyl)-4-aminoglutaric acid) is a naturally occurring, high potency sweetener that was originally isolated from the plant Sclerochiton ilicifolius, found in the Transvaal Region of South Africa. Monatin has the chemical structure: HO O NH 2O OH OH N H [0004] Because of various naming conventions, monatin is also known by a number of alternative chemical names, including but not limited to: 2-hydroxy-2-(indol-3-ylmethyl)-4 aminoglutaric acid; 4-amino-2-hydroxy-2- (1 H-indol-3-ylmethyl)-pentanedioic acid; 4 hydroxy-4-(3-indolylmethyl)glutamic acid; and, 3-(1-amino-1,3-dicarboxy-3-hydroxy-but-4 yl)indole. [0005 Monatin has two chiral centers thus leading to four potential stercoisomeric configurations; the R,R configuration (the "R,R stereoisomer" or "R,R monatin"); the S,S configuration (the "S,S stereoisomer" or "S,S monatin"); the R,S configuration (the "R,S 1 WO 2012/083251 PCT/US2011/065643 stereoisomer" or "R,S monatin"); and the S,R configuration (the "S,R stereoisomer" or "S,R monatin"). [0006] Monatin has an excellent sweetness quality, and depending on a particular composition, monatin may be several hundred times sweeter than sucrose and in some cases thousands of times sweeter than sucrose. Monatin has four stereoisomeric configurations which exhibit differing levels of sweetness. The R,R stereoisomer of monatin is about 2,000-3,000 times sweeter than sucrose by weight. Depending on the methods of manufacture, purification, and intended uses, a monatin composition may include a pure stereoisomer or it may include a mixture of stereoisomers. [0007] Monatin can be isolated from the bark of the roots of the plant Sclerochiton ilicfolius. For example, the bark can be ground and extracted with water, filtered and freeze dried to obtain a dark brown, amorphous mass. The mass can be re-dissolved in water and reacted with a cation resin in the acid form, e.g., AG 50W-X8 in the HCl form (Bio-Rad Laboratories, Richmond, Calif.). The resin can be washed with water and the compounds bound to the resin elated using an aqueous ammonia solution, The eluate can be freeze dried and subjected to aqueous gel filtration. See, e.g., U.S. Pat. No. 5,128,164, which is hereby incorporated by reference in its entirety. Alternatively, monatin can be chemically synthesized. See, e.g., the methods of Holzapfel and Olivier, Synh. Common. 23:2511 (1993); Holzapfel et al., Synlh. Cowmun. 38:7025 (1994); U.S. Pat. No. 5,128,164; U.S. Pat. No. 4,975,298; and U.S. Pat. No. 5,994,559, which are hereby incorporated by reference in their entirety. [0008] Monatin can also be produced via enzymatic processes. Reference is made to WO 2003/091396 A2, which discloses, inter alia, polypeptides, pathways, and microorganisms for in vivo and in vitro production of monatin. WO 2003/091396 A2 (see, e.g., Figures 1-3 and 11-13) and U.S. Patent Publication No. 2005/282260 describe the production of monatin from tryptophan through multi-step pathways involving biological conversions with polypeptides (proteins) or enzymes and are hereby incorporated by reference in their entirety. One pathway described involves converting tryptophan to indole-3-pyruvate ("I-3-P"), converting indole-3-pyruvate to 2-hydroxy 2-(indol-3-ylmethyl)-4-keto glutaric acid (monatin precursor, "MP"), and converting MP to monatin, biologically, for example, with enzymes. This process may be used to make stereoisomer enriched monatin compositions including a monatin composition that comprises greater than 90% R,R monatin. Stevia is a natural sweetener, derived from the South American perennial Stevia rebaudiona. The 2 WO 2012/083251 PCT/US20111/065643 sweetness in the stevia plant is attributed to the presence of high potency sweet glycosides. The two major glycosides found in the leaves and stems of the stevia plant are stevioside and rebaudioside A. Rebaudioside A is a desirable glycoside for use in non-caloric sweeteners. It is recognized that other steviol glycosides or a combination of steviol glycosides may be used as a sweetener. Various methods are available for the purification of rebaudioside A from crude rebaudioside A-containing stevia extracts. [0009] The use of high-potency sweeteners allows for the formulation of sweetened beverages with zero, or at least significantly fewer, calories than standard sweetened beverages, an important health and wellness feature as many countries are attempting to address weight-related public health concerns, In particular, a naturally occurring high potency sweetener with a pleasing, sugar-like taste profile, such as monatin, is desirable. However, at times various high-potency sweeteners, including monatin, may present formulation challenges due to varying chemical stability tinder common beverage conditions, such as low pH, exposure to light, presence of carbonation ("C0 2 "), presence of oxygen ("02"), interaction with various flavor components, packaging that is transparent to ultraviolet ("UV") light or visible light or packaging that is to some extent 02 permeable. These stability challenges can manifest themselves in a perceived loss of sweetness, discoloration, or undesirable flavors or aromas. High-potency sweeteners often affect sensory properties beyond simply maximum sweetness intensity. For example, some high potency sweeteners cause a prolonged sweetness in the mouth, which is not common with caloric sweeteners. Finally, the combination of sweeteners may not behave in predictable ways with respect to sweetness intensity. Selection of appropriate blends of sweeteners may permit more cost-effective blends. The present disclosure addresses one or more of these commercially-relevant concerns, SUMMARY [0010] Provided herein are compositions containing monatin and steviol glyclosides. In one aspect, provided is a method of improving a sweetener performance in a food product, the method comprising: adding monatin or a salt thereof to the food product ; and adding steviol glycosides to the food product, wherein the steviol glycosides are added in an amount sufficient such that the food product exhibits at least one of a reduction in discoloration and a reduction in off-flavors after exposure to ultraviolet or visible light, as compared to a comparable food product containing monatin without steviol glycosides. In some 3 WO 2012/083251 PCT/US20111/065643 embodiments, the food product comprises between about 10 ppm and about 50 ppm of monatin and between about 10 ppm and about 80 ppm of rebaudioside A. In some embodiments, the food product contains less than about 0.5 ppm of 3-methyl indole after 28 days storage at about 22*C in the continuous presence of fluorescent lighting of about 4,000 lux. In some embodiments, the food product retains at least about 65% of its monatin equivalents after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4,000 lux, wherein monatin equivalents are defined as monatin and monatin lactone. In some embodiments, the food product does not exhibit any appreciable discoloration or appreciable off-flavors after 28 days storage at about 22*C in the continuous presence of fluorescent lighting of about 4,000 lux. In some embodiments, the food product exhibits decreased sweetness linger as compared to a comparable food product containing either monatin or steviol glycosides alone. In some embodiments, the food product exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation. In some embodiments, the food product is a beverage. [0011] In a second aspect, provided is a method of making a stable beverage, the method comprising: adding monatin or a salt thereof to the beverage; and adding steviol glycosides to the beverage in an amount sufficient such that the beverage retains at least about 65% of its monatin equivalents after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4000 lux, wherein monatin equivalents are defined as nmonatin and monatin lactone. In some embodiments, the beverage retains at least about 80% of its nionatin equivalents after 28 days storage at about 224C in the continuous presence of fluorescent lighting of about 4,000 lux. In some embodiments, the beverage is a carbonated beverage. In some embodiments, the beverage is a still beverage. In some embodiments, the beverage comprises about 10 ppm to about 80 ppm of rebaudioside A. In some embodiments, the beverage comprises about 10 ppm to about 50 ppm of monatin, In some embodiments, the beverage exhibits decreased sweetness linger as compared to a comparable food product containing either monatin or steviol glycosides alone. In some embodiments, the beverage exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation. [0012] In a third aspect, provided is a beverage comprising: about 10 ppm to about 50 ppm of monatin or a salt thereof; and about 10 ppm to about 80 ppm of steviol glycosides, wherein the beverage retains at least about 65% of its monatin equivalents after 28 days storage at about 22*C in the continuous presence of fluorescent lighting of about 4,000 lux, wherein 4 WO 2012/083251 PCT/US20111/065643 monatin equivalents are defined as monatin and monatin lactone. In some embodiments, the monatin is at least about 90% R,R-monatin. In some embodiments, the monatin is at least about 95% R,R-monatin. In some embodiments, the beverage retains at least about 80% of its monatin equivalents after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4,000 lux. In some embodiments, the beverage is a carbonated beverage. In some embodiments, the steviol glycosides are at least about 95 wt% rebaudioside A. In some embodiments, the steviol glycosides are chosen from the group consisting of rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside F, rebaudioside F, stevioside, dulcoside A, steviolbioside, and a combination thereof. In some embodiments, the beverage exhibits decreased sweetness linger as compared to a comparable food product containing monatin without steviol glycosides. In some embodiments, the beverage exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation. [0013] In a fourth aspect, provided is a beverage composition comprising: about 10 ppm to about 50 ppm of monatin or a salt thereof; and about 10 ppm to about 80 ppm of steviol glycosides, wherein the beverage composition does not exhibit any appreciable odors or any appreciable off-flavors after 28 days storage at 22 C in the continuous presence of fluorescent lighting of about 4,000 lux. In some embodiments, the beverage composition is a carbonated beverage. In some embodiments, the monatin comprises at least about 90% R,R monatin. In some embodiments, the steviol glycosides include rebaudioside A. In some embodiments, the steviol glycosides are at least about 80 wt% rebaudioside A. In some embodiments, the steviol glycosides are at least about 95 wt% rebaudioside A. In some embodiments, the steviol glycosides are chosen from the group consisting of rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, stevioside, dulcoside A, steviolbioside, and a combination thereof. In some embodiments, the beverage exhibits decreased sweetness linger as compared to a comparable food product containing monatin without steviol glycosides. In some embodiments, the beverage exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation. [0014] In a fifth aspect, provided is a beverage composition comprising: about 10 ppm to about 50 ppm of monatin or a salt thereof; and about 10 ppm to about 80 ppm of steviol glycosides, wherein the beverage does not exhibit any appreciable discoloration after 28 days storage at 22 *C in the continuous presence of fluorescent lighting of about 4,000 lux. In 5 WO 2012/083251 PCT/US20111/065643 some embodiments, the beverage composition is a carbonated beverage. In some embodiments, the monatin comprises at least about 90% R,R-monatin. In some embodiments, the steviol glycocides include rebaudioside A. In some embodiments, the steviol glycosides are at least about 80 wt% rebaudioside A. In some embodiments, the steviol glycosides are at least about 95 wt% rebaudioside A. In some embodiments, the steviol glycosides are chosen from the group consisting of rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, stevioside, dulcoside A, steviolbioside, and a combination thereof. In some embodiments, the beverage exhibits decreased sweetness linger as compared to a comparable food product containing monatin without steviol glycosides. In some embodiments, the beverage exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a sweetness intensity versus time curve for various blends of monatin and 95wt% rebaudioside A, as compared to a monatin-only sample, a 95 wt% rebaudioside A-only sample, and a sucrose reference sample. DETAILED DESCRIPTION [0015] As stated above, monatin has two chiral centers leading to four potential stereoisomeric configurations. As used herein, unless otherwise indicated, the term "monatin" is used to refer to compositions including any combination of the four stereoisomers of monatin (or any of the salts thereof), including a single isomeric form. [0016] Unless otherwise stated, the term "monatin" includes any salt thereof. As used herein, unless otherwise stated, the term "monatin" is independent of the method by which the monatin was made, and thus encompasses monatin that was, for example, synthesized in whole or in part by biosynthetic pathway(s), purely synthetic means, or isolated from a natural source. [0017] As used herein, "sucrose equivalence value", or "% SEV", is a measure of sweetness. The SEV is the concentration (% w/v) of a sucrose solution that gives a degree of sweetness that is perceived to be the same degree of sweetness as that of a test sample. The % SEV of an aqueous solution containing monatin is determined through comparison of the monatin containing solution to a series of sucrose-containing solutions prepared in the same medium. The monatin-containing solution may also include a steviol glycoside, which may contribute sweetness to the solution. For purposes of this disclosure, an approximate % SEV can be 6 WO 2012/083251 PCT/US20111/065643 calculated for a solution containing monatin and/or steviol glycosides, based on the concentration response curves for the monatin and/or steviol glycosides in the composition. [0018] Monatin-containing compositions are described herein as being "X" times as sweet as sucrose. The various stereoisomers of monatin are also described as being "X" times sweeter than sucrose. These values are determined from the % SEV of the compositions. [0019] As used herein, the term "about" encompasses the range of experimental error that occurs in any measurement. [0020] In some cases, a "comparable food product" is used herein to compare a monatin containing food product to essentially the same monatin-containing food product that also includes steviol glycosides. Essentially the same food product means that the food product has essentially the same attributes, properties, and/or composition as the food product it is being compared to, and has been exposed to the same conditions as the food product it is being compared to. In some cases, a "comparable beverage" is used herein to compare a monatin- containing beverage to essentially the same monatin-containing beverage that also includes steviol glycosides. As used herein, "food", "food product", "foodstuff", and "food composition" include beverages. [0021] In the process of formulating monatin into various beverage compositions it has been observed that monatin alone is insufficiently stable in extended trials when found in the combined presence of 02 and light (either visible and/or UV). What constitutes appropriate levels of stability varies by specific beverage application and market requirements. A general rule of thumb is that 13 weeks of storage stability (or "shelf stability") under typical store handling conditions is required for common cold-filled consumer beverage applications. And another general rule of thumb is that 26 to 52 weeks of storage stability (or "shelf stability") under typical handling conditions is required for common hot-filled consumer beverage applications. In general, storage stability is not readily quantifiable, but is rather an assessment of organoleptic taste, aroma perception, or appearance. Key features of instability can be identified and used to predict unacceptable levels of shelf stability. In the case of monatin, the levels of total indole-containing molecules (as an indole ring is a substantial portion of the monatin molecule) are a good indicator of the relative shelf stability of various formulations. For purposes of this report, total indole means the sum of monatin, and its lactone and lactam forms. It has also been found that monitoring "musty" off flavors or aromas by trained human sensory evaluators also is predictive of monatin's shelf stability. 7 WO 2012/083251 PCT/US20111/065643 Further, it is believed that a degradation product from monatin that contributes to this "musty" organoleptic property is 3-methyl indole. Limiting the presence of these components in the beverage in some instances can be used to ensure appropriate shelf stability. In particular, limitation of the presence of 3-methyl indole to less than 0.5 ppb is desirable in some embodiments. [0022] In clear or lightly colored beverages, one may also quantitatively observe yellow discoloration. The prevention of discoloration is then predictive of appropriate shelf stability. [0023] It has been discovered that monatin is subject to degradation in the combined presence of dissolved 02 and light resulting in one or more of a loss of sweetness, yellow discoloration, and development of off-flavors or aromas. This light may be in either the visible or UV spectrum. [0024] For purposes of this disclosure, an "off flavor" is defined as a sensory experience of a volatile compound(s) that may be detected orthonasally and/or retronasally. In many instances when an off flavor is referred to herein, the off flavor is a musty off flavor. However, it is recognized that other negative off flavors may be predictive of monatin's shelf stability. Off flavors and discoloration may be referred to herein in terms of an "appreciable" off flavor or an "appreciable" discoloration. For purposes of this disclosure, an appreciable off flavor is detectable nasally by sniffing or retronasally by tasting by a trained human sensory evaluator and an appreciable discoloration is detectable visually by a trained human sensory evaluator. [0025] As stated above, a musty off flavor is believed to be, in part, from formation of 3 methyl indole from monatin. Thus instability may lead to a loss of monatin in a sweetened beverage composition, thus affecting the sweetness properties of the beverage. [0026] The inventors unexpectedly observed that a combination of monatin and steviol glycosides in a beverage resulted in the beverage having an improved sweetener performance, as compared to a beverage containing monatin without steviol glycosides. As described herein, the sweetener performance of a beverage may be defined in terms of its stability, or more specifically, in terms of the absence of off flavors and yellow discoloration after exposure to light over a certain period of time. Sweetener performance may also be described in terms of the preservation or retention of the monatin in the beverage after exposure to light over a certain period of time. Alternatively, sweetener performance of a beverage containing a blend of monatin and steviol glycosides may be described herein in 8 WO 2012/083251 PCT/US20111/065643 terms of having a reduction in discoloration and/or a reduction in off flavors over a certain period of time as compared to a beverage containing monatin without steviol glycosides. The sweetener performance in a beverage may, in some cases, also be defined herein in terms of its sweetness linger. For purposes of this disclosure, sweetness linger is detectable by tasting by a trained human sensory evaluator. The sweetener performance in a beverage may, in some cases, also be defined in terms of its sweetness synergy. For purposes of this disclosure, sweetness synergy is detectable by tasting by a trained human sensory evaluator. [0027] The disclosure herein focuses on compositions containing a blend of monatin and steviol glycosides. The term "steviol glycosides" as used herein refers to any of the glycosides of the aglycone steviol (ent-] 3-hydroxykaur-1 6-en-I 9-oic-acid) including, but not limited to, stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, dulcoside, rebusoside, steviolmonoside, steviolbioside, and 19-0-P glucopyranosol-steviol. As used herein, the term "steviol glycosides" also includes isomers and derivatives of steviol glycosides. "Reb A", as used herein, refers to a stevia leaf derived composition comprising greater than about 95% rebaudioside A. "RA80" refers to a stevia leaf-derived composition comprising greater than about 80% rebaudioside A. [0028] Several grades of steviol glycosides are available. Grades having a high rebaudioside A concentration contribute lower bitter off taste levels. For the compositions disclosed herein, in some embodiments, the steviol glycosides have a rebaudioside A concentration of about 30 to 99.5 wt. % relative to all steviol glycosides, In some embodiments, the steviol glycosides have a rebaudioside A concentration of about 60 to 99.5 wt. % relative to all steviol glycosides. In other embodiments, the steviol glycosides have a rebaudioside A concentration of about 80 to 99.5 wt. % relative to all steviol glycosides. In other embodiments, the steviol glycosides have a rebaudioside A concentration of about 95 to 99.5 wt. % relative to all steviol glycosides, and in yet other embodiments, the steviol glycosides have a rebaudioside A concentration of about 97 to 99.5 wt. % relative to all steviol glycosides. In some embodiments, the steviol glycosides in the blend may be essentially all or predominantly rebaudioside A (predominantly being defined herein as greater than 90% by weight), In some embodiments, the steviol glycosides may include rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, stevioside, dulcoside A, steviolbioside, and a combination thereof. In some embodiments, the steviol glycosides may be predominantly stevioside. In other 9 WO 2012/083251 PCT/US20111/065643 embodiments, the steviol glycosides may be predominantly rebaudioside D. In yet other embodiments, a mixture of any or all of the steviol glycosides may be used. [0029] Methods to selectively extract one or more of the steviol glycosides, for example, rebaudioside A, have been previously disclosed in the art. For example, Japanese Patent 63173531, which is hereby incorporated by reference in its entirety, describes a method of extracting sweet glycosides from the Stevia rebaudiana plant. This procedure isolates a mixture of sweet glycosides. Other techniques include those reported in Japanese Publication Numbers 56121454, 52062300, and 56121453 assigned to Ajinomoto Company, Inc. which are hereby incorporated by reference in their entirety. [0030] Although the disclosure herein is focused on a beverage composition containing monatin and steviol glycosides, it is recognized that this disclosure is applicable to food and beverage compositions. [0031] A composition may be described herein in terms of its total indole concentration, which is defined as monatin and its intramolecular rearrangement products - monatin lactone and monatin lactam. A composition may also be described herein as retaining a certain percentage of monatin equivalents, which is defined herein as monatin plus monatin lactone. [0032] As exemplified in the examples below, in some embodiments, the addition of steviol glycosides to a monatin-containing composition can result in the composition retaining a larger percentage of monatin equivalents after exposure to light as compared to a sample containing monatin without steviol glycosides. Due to the higher percentage of monatin retained, it is believed that there is less yellow discoloration and less conversion to 3-methyl indole, which may be a significant contributor to a musty off flavor. While not being bound by theory, it is believed that steviol glycosides may inhibit monatin photooxidation. [0033] In some embodiments, a food product (e.g., a beverage) including monatin and steviol glycosides may exhibit decreased sweetness linger as compared to a comparable food product containing monatin or steviol glycosides alone [0034] In some embodiments, a food product (e.g., a beverage) including monatin and steviol glycosides may exhibit greater sweetness than the sweetness expected from sweetness calculations made Lising the Beidler equation, i.e, sweetness synergy. The concentration of each sweetener correlating to a target sucrose equivalent value ("% SEV") may be calculated 10 WO 2012/083251 PCT/US20111/065643 from the Beidler curve of concentration response data for each of the individual sweeteners. The standard form of the Beidler equation is: (a * C) Sweetness (b + C) where C is the concentration of the sweetener (in mg/L), and a and b are coefficients calculated from the empirical measurements. Monatin has a different Beidler equation at neutral pH as compared to acid pH. [0035] A beverage composition disclosed herein can include a blend of monatin and steviol glycosides. In some embodiments, the amount of steviol glycosides is equal to or greater than an amount sufficient to cause the beverage composition to exhibit a reduction in discoloration and/or a reduction in off flavors after exposure to ultraviolet and visible light, as compared to a comparable beverage composition containing monatin without steviol glycosides. It is recognized that if steviol glycosides are added to the monatin containing composition, the resulting composition will have a higher sucrose equivalence value ("% SEV"). In order for the composition containing monatin and steviol glycosides to have an essentially equal % SEV to the monatin only containing composition, it is recognized that a smaller amount of monatin would be used in order to account for the sweetness from the steviol glycosides. [0036] In some embodiments, the amount of steviol glycosides is equal to or greater than an amount sufficient such that the beverage retains at least about 65% of its monatin equivalents after 28 days stored at about 224C in the continuous presence of fluorescent lighting of about 4000 lux. In other embodiments, the amount of steviol glycosides is equal to or greater than an amount sufficient such that the beverage retains at least about 80% of its monatin equivalents under the same conditions. It is recognized that other conditions may be used to define a certain percent retention of monatin equivalents. [0037] In some embodiments, beverage compositions containing monatin and steviol glycosides may comprise other stability enhancing features, such as, for example, antioxidants. Reference is made to PCT Publication Number WO 2010/138513, titled "SHELF STABLE MONATIN SWEETENED BEVERAGE," which is hereby incorporated by reference in its entirety. [0038] For a beverage composition having a blend of monatin and steviol glycosides, in some embodiments. monatin is present in an amount that ranges from about 0.0003 to about 1% of 11 WO 2012/083251 PCT/US20111/065643 the beverage composition (i.e., about 3 to about 10,000 ppm) (e.g., about 0.0005 to about 0.2%), including any particular value within that range (e.g., 0.0003%, 0.005%, 0.06% or 0.2% of the beverage composition). For example, a beverage composition may comprise about 0.0005 to about 0.005% (e.g., about 0.001 to about 0.0050%) of the R,R monatin, or about 0.005 to about 0.2% (e.g., about 0.01 to about 0.175%) of S,S monatin. In some embodiments, a beverage composition may comprise about 0.001% to about 0.0050% (ie. about 10 to about 50 ppm), about 0.001% to about 0.0040% (i.e. about 10 to about 40 ppm), or about 0.001% to about 0.0035% (i.e. about 10 to about 35 ppm) of R,R monatin and about 0.001% to about 0.008% (i.e. about 10 to about 80 ppm), about 0.0015% to about 0.0075% (i.e. about 15 to about 75 ppm), or about 0.002% to about 0.007% (i.e. about 20 to about 70 ppm) of Reb A. In some embodiments, the rebaudioside A may be 95 wt% rebaudioside A. In other embodiments, beverages containing monatin and steviol glycosides may have amounts of monatin and/or amounts of steviol glycosides that are outside of the ranges disclosed herein. [0039] It is recognized that an amount of monatin in a beverage composition may vary as a function, for example, of the desired sweetness intensity, the beverage formulation, and the stereoisomeric configuration of the monatin. Similarly, it is recognized that the amounts of steviol glycosides in a beverage composition may vary as a function, for example, of the profile of the steviol glycosides (ie. rebaudioside A, rebaudioside D, stevioside, other steviol glycosides, and any combination thereof), the amount of monatin in the beverage, the desired stability profile, and the presence of any other stability enhancing features. [0040] As used herein, "beverage composition" refers to a composition that is drinkable as is (i.e., does not need to be diluted, or is "ready-to-drink") or a liquid concentrate or a dry powder that can be diluted or mixed with additional liquid to form a drinkable beverage. Beverage compositions herein include carbonated and non-carbonated soft drinks, coffee beverages, tea beverages, dairy beverages, liquid concentrates, flavored waters, enhanced waters, fruit juice and fruit juice-flavored drinks, sport drinks, and alcohol products. Beverages herein include beverages formed from a powder. In some embodiments, beverage compositions comprising monatin and steviol glycosides include other ingredients, such as, for example, a flavoring, coloring, organic acids, inorganic acids, preservatives, caffeine, other sweeteners, and/or polyols. [0041] In some embodiments, the monatin and steviol glycosides containing beverage compositions comprise monatin that consists essentially of SS or RR monatin. In other 12 WO 2012/083251 PCT/US20111/065643 embodiments, the compositions contain predominantly S,S or R,R monatin. "Predominantly" means that of the monatin stereoisomers present in the composition, the monatin contains greater than 90% of a particular stereoisomer. In some embodiments, the compositions are substantially free of S,S or R,R monatin. "Substantially free" means that of the monatin stereoisomers present in the composition, the composition contains less than 2% of a particular stereoisomer. In another aspect, a beverage composition includes a stereoisomerically-enriched monatin mixture. "Stereoisormerically-enriched ionatin mixture" means that the mixture contains more than one monatin stereoisomer and at least 60% of the monatin stereoisomers in the mixture is a particular stereoisomer, such as R,R, S,S, S,R or R,S. In other embodiments, the mixture contains greater than about 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% of a particular monatin stercoisomer. In another embodiment, a beverage composition comprises a stereoisomerically-enriched RR or S,S monatin. "Stercoisomerically-enriched" R,R monatin means that the monatin comprises at least 60% R,R monatin. "Stercoisomerically-enriched" S,S monatin means that the monatin comprises at least 60% S,S monatin. In other embodiments, "stereoisomerically-enriched" monatin comprises greater than about 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% of R,R or S,S monatin. [0042] It should be understood that beverages and other beverage products in accordance with this disclosure may have any of numerous different specific formulations or constitutions. The formulation of a beverage product in accordance with this disclosure can vary to a certain extent, depending upon such factors as the product's intended market segment, its desired nutritional characteristics, flavor profile and the like. For example, it will generally be an option to add further ingredients to the formulation of a particular beverage embodiment, including any of the beverage formulations described below. Additional (ite.. more and/or other) sweeteners may be added, flavorings, electrolytes, vitamins, fruit juices or other fruit products, tastants, masking agents and the like, flavor enhancers, and/or carbonation typically can be added to any such formulations to vary the taste, mouthfeel, nutritional characteristics, etc. In general, a beverage in accordance with this disclosure typically comprises at least water, sweetener, acidulant, and flavoring. Exemplary flavorings which may be suitable for at least certain formulations in accordance with this disclosure include, without limitation, cola flavoring, citrus flavoring, root beer flavoring, and spice flavorings. Carbonation, in the form of carbon dioxide, may be added for effervescence. Natural and synthetic preservatives can be added if desired, depending 13 WO 2012/083251 PCT/US20111/065643 upon the other ingredients, production technique, desired shelf life, etc. Optionally, caffeine can be added. Certain exemplary embodiments of the beverages disclosed herein are, without limitation, cola-flavored carbonated beverages, characteristically containing carbonated water, sweetener, kola nut extract, and/or other cola flavoring, caramel coloring, and optionally other ingredients. Additional and alternative suitable ingredients will be recognized by those skilled in the art given the benefit of this disclosure. [0043] Some embodiments may be considered still beverages, i.e., beverages which are not carbonated. Common examples include, without limitation, coffee beverages, tea beverages, dairy beverages, flavored waters, enhanced waters, non-carbonated soft drinks, fruit juice and fruit juice-flavored drinks, sport drinks, and alcohol products other than beer and champagnes. [0044] In other embodiments, the beverage may contain dissolved carbon dioxide ("C0 2 ") in amounts sufficient to provide effervescence. Common examples include, without limitation, carbonated soft drinks, beer and champagnes. Such carbonated beverages typically have carbon dioxide concentrations of about 1.6 volumes CO 2 per volume of beverage to about 4.2 volumes CO 2 per volume of beverage. Carbon dioxide is typically introduced into a beverage by either fermentation (as in the case of beer and champagnes) or dissolving the carbon dioxide into the beverage under pressure (as in the case of carbonated beverages). Specific methods of beverage carbonation are well known to those skilled in the art. [0045] In carbonated beverages, the process of carbonation results in a removal or displacement of dissolved oxygen from the beverage, In some embodiments, the dissolved oxygen levels may be reduced or controlled for stability purposes, as disclosed in WO 2010/138513, which is hereby incorporated by reference in its entirety. 14 WO 2012/083251 PCT/US20111/065643 EXAMPLES [0046] Aspects of certain embodiments in accordance with aspects of the invention are illustrated in the following examples. The materials and methods described in these examples are illustrative and not intended to be limiting. [0047] Three lots of monatin were used in the following examples: Lot #5837-72RRSYN monopotassium salt; Lot #25001119 (90.3% 2R,4R- monatin potassium/sodium salt); Lot #25001392 (90.4% 2R,4R- monatin sodium salt). Monatin concentration is expressed on a pure monatin salt basis in the examples unless otherwise specified. Lot #5837-72RRSYN was used in Example I1. Lot #25001119 was used in Examples I through 6 and 8 through 10. Lot #25001392 was used in Examples 7 and 12 through 16. [0048] The Beidler equation for monatin in water is % SEV = 26.7 * [monatinj/(69,6 + [monatin]). The Beidler equation for Reb A in water at room temperature is % SEV = 1 L26*[Reb A]/(253.7 +_[Reb A]), These equations were used in all examples unless otherwise specified. Example 1 [0049] Lemon-lime flavored carbonated beverages were stored in the dark and evaluated over a seven week period. Solutions of about 9 mM citric acid/trisodium citrate (pH1 of approximately 3.0) in high purity water were sweetened to about 8.0% SEV (the calculated % SEV of each sample is shown in Table I below) with various amounts of monatin and/or Reb A, depending on the intended blend, and then flavored with lemon-lime flavor; and preserved with approximately 150 ppm sodium benzoate. The solutions were carbonated in 10 liter batch sizes using an Armfield carbonator, by adding approximately 3.5 liters of carbon dioxide ("CO2") per liter of solution before being transferred to twelve ounce clear, glass bottles. Caps were placed on the bottles after filling. The bottles were stored in a dark oven maintained at a temperature of approximately 38*C. Multiple bottles of each sample were created since a bottle was sacrificed when the measurements below were taken, [0050] Total indole (monatin and its intramolecular rearrangement products, monatin lactone and monatin lactam) concentrations were quantified by with a Waters Alliance 2690 H PLC at 40'C using a Zorbax Eclipse XDB-C18 column (Agilent Technologies, Inc., Santa Clara CA) with UV detection at 280 nm by a photodiode array detector. The compounds were eluted by a gradient mobile phase consisting of (A) 0.3% formic acid in water with 10 mM ammonium formate and (B) 1:1 methanol and acetonitrile (v:v) with 0.3% formic acid and 10 mM 15 WO 2012/083251 PCT/US20111/065643 ammonium formate. The integrated peak areas for the three compounds were expressed in terms of monatin concentration (mg/L) based on a standard curve prepared for monatin. Thus, the extinction coefficient for monatin was applied to estimate concentrations of monatin lactone and monatin lactarn since standards for these compounds were unavailable. [0051] Total indole concentrations are shown in Table I for each of the blends. The percent of the original indole concentration is shown in parentheses. Table 2 shows monatin equivalents, which is defined for purposes herein as monatin and monatin lactone; the percent of the original monatin equivalents is shown in parentheses. Table 1. Total Indole Concentrations for Various Blends of Monatin and Reb A Stored in the Dark Sample 1 2 3 4 5 Monatin (% SEV) 7.9 6.3 4.8 3.4 1.7 Reb A(% SEV) 0.0 1.6 3.2 4.8 6.4 Total (% SEV) 7.9 7.9 8.0 8.2 8.1 Rob A (ppm) 0 35 81 144 236 Time (Weeks) Total Indole (ppm) 0.0 35.6 (100,0) 23.6 (100.0) 15.8 (100.0) 10.0 (100.0) 4.5 (100.0) 2.0 35.6 (100.0) 23.6 (100.0) 15.8 (100.0) 10.0 (100.0) 4.5 (100.0) 4.0 32.8 (92.1) 22.2 (94.1) 14.9 (94.3) 9.4 (94.0) 4.1 (91.1) 7.3 33.0 (92.7) 23.9 (101.3) 16.0 (101.3) 9,3 (93.0) 4.4 (97.8) Table 2. Monatin Equivalents Concentration for Various Blends of Monatin and Reb A Stored in the Dark Sample Time (Weeks) 1 2 3 4 5 0.0 35.6 (100.0) 23.6 (100.0) 15.8 (100.0) 10.0 (100.0) 4.5 (100.0) 2.0 31.2 (87.6) 20.6 (87.3) 13.8 (87.3) 8.8 (88.0) 3.9 (86.7) 4.0 25.1 (70.5) 16.6 (70.3) 11.1 (70.3) 7.1 (71.0) 2.9 (64.4) 7.3 20.3 (57.0) 14.8(62.7) 9.7(61.4) 5.6(56.0) 2,7 (60.0) 16 WO 2012/083251 PCT/US20111/065643 Table 3. Lactan Concentration for Various Blends of Monatin and Reb A Stored in the Dark Sample Time (Weeks) 1 2 3 4 5 0.0 0.0 (0.0) 0.0 (0.0) 0.0(0.0) 0.0(0.0) 0.0 (0.0) 2.0 4.4 (12.4 .0 (0.) . (1 .0) .0 (12.0) 0.6 (13.3) 4.0 7.7 (23.5) 5.6 (25.2) 3.8 (25.5) 2.3 (24.5) 1.2 (29.7) 7.3 12.7 (38.5) 9.2 (38.5) 6.3 (39.4) 3.6 (38.7) 1.7 (38.6) [0052] Table 2 shows that there is a decrease in monatin equivalents (monatin plus monatin lactone) over time. The decrease in monatin equivalents may be attributed to the formation of monatin lactam from monatin. The presence of Reb A in samples 2 through 5 did not affect the proportion of monatin converted to monatin lactam. Example 2 10053] Compositions from Example I were freshly prepared and exposed to light. The solutions were prepared using the same steps described above for Example 1. The bottles containing the solutions were exposed to light in a light box (approximately 4,000 lux light intensity) continuously (24 hours per day), using a rotating carousel to provide even illumination. The source of light was provided by ultra-violet ("UV") light wavelength transmitting bulbs and standard fluorescent bulbs. The heat produced by the lights warmed the samples to a temperature of about 22 0 C. Multiple bottles of each sample were created since each bottle was sacrificed when the measurements below were taken. The samples were analyzed for total indole concentration and monatin equivalents, using the methodology disclosed in Example I above. 17 WO 2012/083251 PCT/US20111/065643 Table 4. Total Indole Concentrations for Various Blends of Monatin and Reb A Exposed to Light Sample 1 2 3 4 5 Monatin (% SEV) 7.9 6.3 4.8 3,4 1.7 Reb A (% SEV) 0.0 1.6 3.2 4.8 6.4 Total (% SEV) 7.9 7.9 8.0 8,2 8.1 Reb A (ppm) 0 35 81 J 144 236 Time (Weeks) Total Indole (ppm) 35.6 0.0 (100.0) 23.6 (100.0) 15.8 (100.0) 10.0 (100.0) 4.5 (100.0) 2.3 32.7 (92.0) 23.1 (97.9) 18.1 (114.3) 9.3 (93.7) 4.2 (93.0) 4.4 26.9 (75.7) 20.4 (86.4) 11.8 (74.8) 8.2 (82.2) 3.7 (82.5) 7.3 22.5 (63.2) 20.6 (87.5) 7.9 (50.1) 7.9 (79.7) 3,7 (81.8) Table 5. Monatin Equivalents for Various Blends of Monatin and Reb A Exposed to Light Sample Time (Weeks) 1 2 3 4 5 35.6 0.0 (100.0) 23.6 (100.0) 15.8 (100.0) 10.0 (100.0) 4.5 (100.0) 2.3 31.8(89.3) 22.4(94.9) 18.0 (113.9) 9.1 (91.0) 4.1 (91.1) 4.4 25.7 (72.2) 19.3 (81.8) 11.4 (72.2) 7.8 (78.0) 3.5 (77.8) 7.3 21.0 (59.0) 19.3 (81.8) 7.2 (45.6) 7.4 (74.0) 3.7 (82.2) [0054] Tables 4 and 5 show that each of the blends of monatin and Reb A, with the exception of sample 3, had a smaller loss of monatin over time, as compared to sample I having monatin alone. Sample 3, which contained an amount of Reb A in between the amount in samples 2 and 4, showed the largest loss of monatin over time compared to all samples, including sample 1. Although a specific explanation is not provided, it is believed that there was a deficiency in sample 3, for example, an error in preparation of the sample. [0055] At the same points in time that data was collected for Tables 4 and 5, the samples were also evaluated for discoloration by comparing each sample to a control solution stored in the same environment but covered in foil and thus protected from light. The evaluator gave the sample a rating between 0 and 9, and the results are shown in Table 6 below. Similarly, the same evaluator observed the samples for any off flavors (using the same 18 WO 2012/083251 PCT/US20111/065643 control used for discoloration), using a scale of 0 and 9, and the results are shown in Table 7 below. Table 6. Discoloration of Blends Exposed to Light Sample 2 3 4 Time aYellow Color Intensity Rating (Weeks) 60 ppm "35 ppm b81 ppm b144 ppn "2 ppm 0.0 0 0 0 0 0 2.3 1 0 0 0 0 4.4 1 0 0 0 0 7.3 J No Sample 1 2 1 0 "The intensity of color was rated on a scale from 0 to 9 where 0 = none, 1 = trace, 2 = faint, 3 = slight, 4 = mild, 5 = moderate, 6 = definite, 7 = strong, 8 = very strong, and 9 = extreme. blInitial Reb A concentration. Table 7. Musty Off-flavor of Blends Exposed to Light Sample 1 2 3 4 5 Time "Musty Off-Flavor Intensity Rating (Weeks) 0 ppm t)35 ppm 8 pp b 144 ppm '236 ppm 0 0 0 0 0 0 2.3 0 0 0 0 0 4.4 0 0 0 0 0 7.3 NoSample 0 2 0 0 "The intensity of musty off-flavor was rated on a scale from 0 to 9 where 0 = none, I trace, 2 = faint, 3 = slight, 4 = mild, 5 = moderate, 6 = definite, 7 = strong, 8 = very strong, and 9 = extreme. bInitial Reb A concentration. [0056] Discoloration was observed in sample I (no Reb A) at 2.3 weeks and 4.4 weeks. However, for samples 2 through 4 having a blend of monatin and Reb A, discoloration was not observed until 7.3 weeks. None of the samples exhibited a musty off flavor except for sample 3 at 7.3 weeks. 19 WO 2012/083251 PCT/US20111/065643 Example 3 [0057] Solutions containing blends of monatin and Reb A were exposed to light and underwent the same evaluations as were done for the samples in Example 2, In contrast to Example 2, the solutions were exposed to sunlight rather than fluorescent light. More specifically, the samples were stored on a windowsill of a north facing window and at a temperature ranging between about 20 and about 25 degrees Celsius. The solutions were prepared in the same manner as those of Examples I and 2 above. [0058] Table 8 shows the various blends of monatin and Reb A. The amount of monatin varied at 13.1 ppm, 17.6 ppm, 21.2 ppm, 25.4 ppm and 31.6 ppm. Except for the sample containing 31.6 ppm monatin, the samples were created by making a composition and dividing it into two equal sized samples, with the first sample containing only monatin and the second sample containing monatin and Reb A. For example, sample I contained 13.1 ppm of monatin for a % SEV of 4.2%; sample 2 contained 13.1 ppm of monatin and 73.2 ppm of Reb A for a % SEV of 7.2%. This sample preparation method reduced the potential for a formulation error and improved the validity of the resulting comparative data between monatin alone and its corresponding monatin/Reb A blend. [0059] Samples 2, 4, 6, and 8 had % SEVs of 7.2%, 7.4%, 7.3%, and 7.3% respectively, and contained a blend of monatin and Reb A. These samples were compared to sample 9 having a % SEV of 7.4% without any Reb A. 20 WO 2012/083251 PCT/US20111/065643 Table 8. Blends of Monatin and Reb A Sample 1 2 3 4 5 6 7 8 9 Monatin 4.2 4.2 5.2 5.2 5.9 5.9 6.6 6.6 7,4 (% SEV) RebA 0 3.0 0 2.2 0 1.4 0 0.7 0 (% SEV) Total 4.2 7.2 5.2 7.4 5.9 7.3 6.6 7.3 7.4 (% SEV) "Monatin (M )nt 13.1 13.1 17.6 17.6 21.2 21.2 25.4 25.4 31.6 (ppmn) "RbA (ppm A 0 73.2 0 50.6 0 29.7 0 14.6 0 (ppmn) 'Initial monatin concentration. bInitial Reb A concentration, [0060] Total indole concentration for each sample was measured at various points using the methodology described above under Example I and is shown in Table 9 below. (The values in parentheses represent the percentage of total indole remaining relative to the initial concentration.) For each comparison, after exposure to sunlight for ten weeks, the sample containing monatin and Reb A had a higher remaining indole concentration as compared to the sample containing monatin alone. Table 10 shows the stability of the Reb A in the blend over time. (The values in parentheses represent the percentage of Reb A remaining relative to the initial concentration.) 21 WO 2012/083251 PCT/US20111/065643 Table 9. Total Indole Concentration (ppm) Sample Time (Weeks) 1 2 3 4 5 6 7 8 9 0 13.1 13.1 17.6 17.6 21.2 21.2 25.4 25.4 31.6 (100.0) (100.0) (100.0) (100.0) (100.0) (100.0) (100.0) (100.0) (100.0) 2 11.3 12.5 15.3 15.6 18.5 19.0 22.6 22.2 26.4 (86.3) (95.4) (86.9) (88.6) (87.3) (89.6) (89.0) (87.4) (83.5) 4 8.8 11.5 14.0 13.3 18.4 18.7 19.8 21.5 17.6 (67.2) (87.8) (79.5) (75.6) (86.8) (88.2) (78.0) (84.6) (55,7) 6 9.2 10.5 10.5 13.2 17.9 19.6 14.9 20.1 20.3 (70.2) (80.2) (59.7) (75.0) (84.4) (92.5) (58.7) (79.1) (64.2) 8 6.2 8.6 6.2 11.3 17.5 20.3 13.1 19.8 5.2 (47.3) (65.6) (35.2) (64.2) (82.5) (95.8) (51.6) (78,0) (16.5) 10 7.6 8.8 4.7 11.5 15.9 20.3 11.7 17.7 10.7 (58.0) (67.2) (26.7) (65.3) (75.0) (95.8) (46.1) (69.7) (33.9) 22 WO 2012/083251 PCT/US2011/065643 Table 10. Reb A Concentration (ppm) Sample (Weeks) 1 2 3 4 5 6 7 8 9 0 N/A 73.2 N/A 50.6 N/A 29.7 N/A 14.6 N/A (100.0) (100.0) (100.0) (100.0) 2 N/A 71.7 N/A 50.2 N/A 27.2 N/A 15.7 N/A (97.3) (99.2) (91.6) (107.5) 4 N/A 72.4 N/A 49.7 N/A 29.6 N/A 15.6 N/A (98.9) (98.2) (99.7) (106.8) 6 N/A 71.9 N/A 49.8 N/A 27.2 N/A 13.9 N/A (98.2) (98.4) (91.6) (95.2) 8 N/A 72.3 N/A 50.8 N/A 28.4 N/A 13.8 N/A (98.8) (100.0) (95.6) (94.5) 10 N/A 72.2 N/A 50.1 N/A 26.9 N/A 13.8 N/A (98.6) (99.0) 90.6) (94.5) [0061] The color of each sample was also measured using a HunterLab ColorQuest XE colorimeter (Hunter Associates Laboratory, Inc., Reston, VA). As shown in Table I 1 below, as the level of monatin in the solution increased, the value of color intensity similarly increased, with the exception of sample 5. The color intensity data also shows that the yellow discoloration was reduced by adding Reb A to the monatin-containing solution. 23 WO 2012/083251 PCT/US20111/065643 Table 11. Yellow Color (b*) Intensity Measured by Colorimeter Sample Time 1 2 (Weeks) 0 0.02 0.01 0.01 0.01 0.02 0.02 0.02 0.02 0.03 2 0.24 0.19 0.37 0.29 0.44 0.27 1.14 0.45 1.17 4 ll 0.42 0.60 0.48 0.54 0.22 1.50 0.60 3.64 6 1.23 0.82 1.65 0.73 0.71 0.18 2.88 0.72 3.34 8 1.96 1.56 3.53 1.36 0.82 0.21 3.54 0.79 7.66 10 1.53 1.42 3.20 1.46 1.08 0.99 3.99 1.03 5.23 [0062] The solutions were also evaluated for the presence of any off flavors. As shown below, for the majority of the samples containing monatin without Reb A, the solutions developed a musty off flavor over time, as compared to the solutions blended with monatin and Reb A, which did not exhibit a musty off flavor after 6 weeks. An off flavor was observed in all of the samples after 8 weeks. However, the intensity of the off flavor was less for the samples containing monatin and Reb A, as compared to the samples containing monatin only. Table 12. Musty Off-Flavor Development (Odor Intensity Score) Sample Time 1 2 3 4 5 6 7 8 9 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 1 0 1 4 0 0 0 0 0 0 1 0 1 6 1 0 1 0 0 0 2 0 2 8 4 2 2 2 2 2 3 3 3 10 4 3 3 2 2 1 3 2 3 The intensity of musty off-flavor was rated on a scale from 0 to 9 where 0 = none, 1 = trace, 2 = faint, 3 = slight, 4 = mild, 5 = moderate, 6 = definite, 7 = strong, 8 = very strong, and 9 = extreme. 24 WO 2012/083251 PCT/US20111/065643 Example 4 [0063] Lemon-lime flavored carbonated soft drinks were made using the formulas shown in the table below. Table 13. Blends of Monatin and Reb A Concentration (% by Weight) Ingredients SampI 1 SamIple 2 Sample 3 Sample 4 Water 99.601 99.599 99.595 99.575 Citric Acid 0.160 0.160 0.160 0.160 Trisodium Citrate 0.020 0.020 0.020 0.020 Sodium Benzoate 0.015 0.015 0.015 0.015 R,R-Monatin Salt 0.0040 0.0027 0.0017 0 Reb A 0.0000 0.0035 0.0081 0.0300 Lemon-lime Flavor 0.200 0.200 0.200 0.200 Total 100.000 100.000 100.000 100.000 [0064] The solutions were evaluated by a six-member sensory panel with experience in evaluating sweetness. A one-ounce sample of each solution, in random order, was tasted cold by each panelist and then rated for lingering sweetness intensity after ten seconds. Some of the panelists spit the sample out; others swallowed the sample - each panelist was consistent with his or her methodology. The panelists arrived at a consensus score for sweetness linger for each sample. Table 14. Sweetness Linger Sample I Sample 2 Sample 3 Sample 4 Total % SEV 8.0 8.0 8.0 8.0 % SEV from Monatin 8.0 6.4 4.8 [ 0 % SEV from Reb A 0 1.6 3.2 8.0 Sweetness Lingering 5 4 3 5 Scored "Sweetness linger was rated on a scale from I to 9 where 1 = Not at all lingering, 3 = Slightly lingering sweetness, 5 = Moderately lingering sweetness, 7 = Very lingering sweetness, and 9 = Extremely lingering sweetness. [0065] The linger scores in Table 14 show that the blends of monatin and Reb A (Samples 2 and 3) had a reduced linger as compared to both the solution containing monatin only (Sample 1) and the solution containing Reb A only (Sample 4). 25 WO 2012/083251 PCT/US20111/065643 Example 5 [0066] The relationship between sweetness intensity and time as it relates to monatin, Reb A, and blends of the two sweeteners was studied. [0067] Samples were prepared to about 8% sucrose equivalent sweetness. The blends were formulated such that the ratio of sweetness from monatin to rebiana was about 100/0, 90/10, 80/20, 70/30, 60/40 and 0/100 (Table 15). The concentration of each sweetener correlating to the target sucrose equivalent value ("% SEV") was calculated from the Beidler curve of concentration response data for each of the individual sweeteners. The standard form of the Beidler equation is: Sweetness = (a (b + C) where C is the concentration of the sweetener (in mg/L), and a and b are coefficients calculated from the empirical measurements. [0068] The Beidler equation for Reb A at room temperature in water is % SEV = (1 .26*C) / (253.7+C) and for monatin at room temperature and pH 3 is % SEV = (16.32* C) / (41.38 + C), where C = concentration of sweetener in mg/L. All samples were prepared in a citrate buffer at pH 3.0 and room temperature. 26 WO 2012/083251 PCT/US20111/065643 Table 15. Formulas for Sample Solutions and Calculated (Theoretical) Sucrose Equivalence Value Monatin: Monatin Reb A Monatin Reb A Total Reb A (ppm) (ppm) (% SEV) (% SEV) (% SEV) Ratio 100/0 43 0 8.3 0 8.3 90/10 36 19 7.5 0.8 8.3 80/20 29 42 6.8 1.6 8.4 70/30 24 69 6.0 2.4 8.4 60/40 19 101 5.2 3.2 8.4 0/100 0 623 0 8.0 8.0 Sucrose N/A N/A N/A N/A N/A (reference) [0069 Table 16 shows the formulation for each of the blends in Table 15, as well as the 8% sucrose reference sample. Table 16. Formulations of the Monatin/Reb A Blends Concentration (% by Weight) Ingredients 100/0* 90/10* 80/20* 70/30* 60/40* 0/100* Sucrose Water 99.816 99.815 99.813 99.810 99.808 99.758 91.82 Citric Acid 0.160 0.160 0.160 0.160 0.160 0.160 0.16 Trisodium 0.020 0.020 0.020 0,020 0.020 0.020 0.02 Citrate R,R- 0.0043 0.0036 0.0029 0.0024 0.0019 0 0 Monatin Salt Reb A 0.0000 0.0019 0.0042 0.0069 0.0101 0.0623 0 Sucrose 0 0 0 0 0 0 8.00 Total 100.000 100.000 100.000 100.000 100.000 100.000 100.000 *Monatin to Reb A ratio 27 WO 2012/083251 PCT/US20111/065643 [0070] A panel of six trained and experienced panelists performed the time intensity evaluation testing. The panelists were instructed to put the entire one fluid ounce sample in his or her mouth and hold for ten seconds while continuously rating the intensity of sweetness, then swallow or spit the sample and continue rating sweetness intensity for one minute. Sweetness intensity was recorded using a computer mouse and Sensory Information Management System ("SIMS") 2000 software (Sensory Computer Systems, Morristown, NJ). The panelists moved the mouse to the right as the sweetness intensity increased and to the left as the intensity decreased. All samples were served at room temperature in a balanced random order, one at a time, and evaluated in duplicate. Panelists took a five-minute break between samples and cleansed their mouth with water and crackers. Results of the time intensity evaluation are shown in Figure 1. [0071] The goal was for all of the samples in the study to have equivalent sweetness. The Reb A sample matched target sweetness of the sucrose reference very closely. After adjusting for R,R-monatin purity, the monatin sample and high-ratio monatin blends were sweeter than the sucrose reference (as outlined in Table 15). Surprisingly, the sweetness of monatin and rebiana blends did not fall somewhere between the 100% monatin and 100% Reb A. Rather, the blends were all sweeter than either monatin or Reb A alone, evidence of an unexpected sweetness synergy between the two sweeteners. Example 6 [0072] Sweetener samples were prepared to about 8 % SEV for quantitative descriptive analysis ("QDA") as described in Example 5, [0073] A panel of nine trained and experienced panelists performed quantitative descriptive analysis ("QDA") on the samples using typical QDA methodology. The panelists tasted a set of control and test samples to develop the ballot with taste attributes, and then were further trained with reference solutions to practice scoring all of the samples on a fifteen-point intensity line scales developed for each attribute. Samples were served at room temperature in a balanced random order, one at a time, and evaluated in duplicate. Panelists had a five minute break between evaluations and used water and crackers for rinsing. Results of the QDA testing are summarized in Table 17. 28 WO 2012/083251 PCT/US20111/065643 Table 17: Quantitative Descriptive Analysis Results Samples Time to Maximum Sour Bitter Metallic Drying Reach Sweetness Astringent Maximum Sweetness 8% Sucrose 1.29 a 7.41 a 4.38 b 1.02 b 0.90 b 3.19 a Monatin 1.57 a 7.27 a 4.90 ab 1.22 b 0.85 b 3.47 a Reb A 1.70 a 6.34 b 5.15 a 2.65 a 1.46 a 3.94 a 9OMonatin/ 1.48 a 7.38 a 4.69 ab 1.14 b 0.94 b 3.48 a 1OReb A Blend 80Monatin/ 1.39 a 7.38 a 4.45 b 1.32 b 0.83 b 3.72 a 20Reb A Blend 70Monatin/ 1.35 a 7.54 a 4.52 ab 1.56 b 0.92 b 4.03 a 30Reb A Blend 60Monatin/ 1.61 a 7.65 a 4.77 ab 1.41 b 0.94 b 3.64 a 40Reb A Blend *Means followed by different letters are significantly different from each other at p <0.05. [0074] The data in Table 17 showed no difference between samples in time to reach maximum sweetness or in drying and astringent attributes. However, it was surprising to observe sweetness synergy between the monatin and Reb A blends, in that that maximum sweetness of the blends was greater than either monatin or Reb A alone. Blends were also less sour than either sweetener alone. Blending with monatin reduced the bitter and metallic attributes most closely attributed to Reb A. Example 7 [0075] The sensory attributes of monatin, Reb A, and monatin/Reb A blends using QDA methodology were evaluated in water. A monatin solution and a Reb A solution were first formulated to match the sweetness of a 7% sucrose solution based on tasting trials by a QDA 29 WO 2012/083251 PCT/US20111/065643 panel. Then the Beidler equations for monatin and Reb A at room temperature, neutral pH were applied to formulate various blends of monatin and Reb A that had equivalent theoretical sweetness as the individual sweeteners (7% SEV). Formulations are outlined in Table 18, Table 18. Test Sample Formulations # Name Monatin Reb A Sucrose Concentration Concentration Concentration I Sucrose 0% 0% 7% 2 Monatin 0.0036% 0% 0% 3 Reb A 0% 0.0417% 0% 4 80Monatin/20Reb A 0.0027% 0.0036% 0% 5 60Monatin/4OReb A 0.0019% 0.0084% 0% 6 40Monatin/60Reb A 0.0012% 0.0151% 0% 7 20Monatin/80Reb A 0.0006% 0.0251% 0% [0076] Nine trained and experienced QDA panelists evaluated samples using typical QDA methodology. They trained with references, identified attributes to put on the ballot, aligned on tasting technique to be used, and practiced scoring the samples using the ballot. Samples were evaluated in individual sensory booths. All samples, except sucrose, were evaluated in a balanced sequential order one at a time. The sucrose sample was evaluated at the end of each session. Panelists were given 7% sucrose solution reference to calibrate scoring before the booth sessions. Samples size was 1.5 fluid ounces presented in a 2 ounce souffl6 cup and samples were presented at room temperature. All samples were evaluated in duplicate. Panelists took a five-minute break between samples and were provided with apples, carrots, and sparking water and crackers to rinse and cleanse their palates during the break. Results of the QDA Analysis are summarized in Tables 19 and 20. 30 WO 2012/083251 PCT/US20111/065643 Table 19. QDA Panel Results - Mean Intensity Scores for Flavor Attributes Time Maximum Herbal Bitterness* Non- Sweet Sucrose Maximum Sweetness* Flavor* Aftertaste Flavors* Sweetness* Immediately** Sucrose 1.1c 6.1 a 0.5 c 0.3 d 0.7 d 5.1 ab Monatin 1.4 c 6.1 ab 0.9 be 0.9 c 2.0 e 4.5 c Reb A 2.0 a 6.3 ab 1.8 a 2.1 a 3.3 a 4.8 be 8OMonatin! 1.4 be 7.0 c 0.8 be 0.8 ed 1.9 c 5.3 ab 20RebA 60Monatin/ 1.3 c 6.9 e 0.8 be 1 1 be 2.4 be 5.5 a 40Reb A 40Monatin/ 1.8 a 6.7 be 1.3 ab 1.2 be 2.6 b 5.1 ab 60Reb A 20Monatin/ 1.7 ab 7.1 e 1.3 ab 1.6 ab 2.5 b 54 a 8OReb A *Means followed by different letters are significantly different from each other at p <0.05. Table 20. Mean Intensity Scores for Aftertaste Attributes (after 30 and 60 Seconds of Swallowing) Samples Sweet Herbal Bitter Non-Sucrose Sweet Drying/ Aftertaste Aftertaste Aftertaste Flavors Aftertaste Astringent 30sec* 30sec 30sec* Aftertaste 60sec Afterfeel 3Osec* 60sec* Sucrose 2.2 b 0.3 a 0.2 b 0.5 d 0.8 a 2.2 c Monatin 2.7 ab 0.4 a 0.8 b 1.7 be 1.6 a 2.9 b Reb A 2.7 ab 0.9 a 1.4 a 2.5 a 1.3 a 3.0 ab 8OMonatin/ 2.9 a 0.3 a 0.5 b 1.4 c 1.4 a 2.9 b 20Reb A 60Monatin/ 3.2 a 0.5 a 0.7 b 1.8 be 1.4 a 2.9 b 40Reb A 40Monatin/ 3,0 a 0.6 a 0.7 b 2.0 ab 1.4 a 3.0 ab 60Reb A 20Monatin/ 3.0 a 0.7 a 1.3 a 2.0ab 1.5 a 3.3 a 8OReb A *Means followed by different letters are significantly different from each other at p 10.05. 31 WO 2012/083251 PCT/US20111/065643 [0077] The data show that there was no significant difference in maximum sweetness between the sucrose, monatin and Reb A samples, which all scored a sweetness intensity score of about 6. All of the monatin/Reb A blends were formulated to be equally sweet to the single sweeteners using data from the individual sweeteners' Beidler equations, but surprisingly, all of the blends were actually significantly sweeter than predicted, demonstrating sweetness synergy between the two sweeteners. 10078] In summary, Reb A sample scored significantly higher in: time to reach max sweetness (slower onset); herbal flavor; bitter taste/aftertaste; and non-sucrose flavors/aftertaste compared to sucrose, monatin, 20Monatin/2OReb A, and 60Monatin/4OReb A samples. The 80Monatin/20Reb A blend scored the closest to sucrose in herbal, bitter, and non-sucrose flavors attributes followed by monatin and 60Monatin/4OReb A blend. The 80Monatin/2OReb A scored closer to sucrose in herbal, bitter and non-sucrose flavor attributes followed by monatin, and 60Monatin/4OReb A blend, Reb A scored the most different in its flavor/aftertaste profile except for sweetness compared to sucrose. Example 8 [00793 Samples of blends were made and tested to further evaluate an effect of Reb A on the linger of monatin. Table 21. Blends of Monatin and Reb A Sample ID Monatin Reb A(ppm) Monatin Reb A Total (ppm) % SEV % SEV Calculated % SEV Sucrose - - - - 8.0 Monatin 40 0 8.0 0.0 8.0 90Mon/lOReb 33 19 7.2 0.8 8.0 8OMon/2OReb 27 42 6.4 1.6 8.0 70Mon/3OReb 22 69 5.6 2.4 8.0 60Mon/4OReb 17 101 4.8 3.2 8.0 32 WO 2012/083251 PCT/US20111/065643 Table 22. Formulations of the Monatin/Reb A Blends of Table 20 Concentration (% by Weight) Ingredients 100/0 90/10 80/20 70/30 60/40 Water 99.819 99.818 99.812 99.812 99.810 Citric Acid 0.160 0.160 0.160 0.160 0.160 Trisodium 0.020 0.020 0.020 0.020 0.020 Citrate R,R-Monatin 0.0040 0.0033 0.0027 0.0022 0.0017 Salt Reb A 0.0000 0.0019 0.0042 0.0069 0.0101 Total 100.000 100.000 100.000 100.000 100.000 [0080] Samples of the solutions were served at ambient temperature (approximately 70 to 72 *F). Six panelists were instructed to put the entire sample (about I ounce) in their mouth, and rate the initial intensity of sweetness using a 9-point intensity scale (0=none and extremely sweet) while holding the sample for 10 seconds and moving their tongue around in their mouth. Each of the panelists then swallowed or spit the sample, and continued rating sweetness intensity of each sample every 10 seconds. All samples were served blind coded in a balanced random order to the panelists. The panelists rated each sample in duplicate. Table 23 shows the mean sweetness intensity of the six panelists at four specific points in time. Table 23. Mean Sweetness Intensity Results Sample Description Initial 1Osec 20sec 30sec Sucrose 7.30 7.05 5.05 3.42 Monatin 7.63 7.35 5.70 3.90 90Mon/lOReb 7.28 6.48 4.97 2.88 8OMon/2OReb 7.76 7.50 5.45 3.69 70Mon/3OReb 7.68 7.26 5.42 3.63 60Mon/4OReb 7.45 7.04 5.39 3.65 [0081] Each panelist recorded the sweetness intensity at 10 second intervals up to 80 seconds. Each panelist's rating for the duplicate samples was then averaged and plotted. The area under the curve (AUC) was calculated for each sample for each panelist. A linger index was calculated by dividing the AUC for each test sample by the AUC for sucrose for that panelist. This compares each panelist's linger response of the blends relative to that panelist's response to sucrose. The linger index for each panelist, as well as the mean value and standard deviation across panelists, is shown below. 33 WO 2012/083251 PCT/US20111/065643 Table 24. Linger Index Values Panelist Panelist Panelist Panelist Panelist Panelist Standard 1 2 3 4 5 6 Mean Deviation Sucrose 1.00 1.00 1.00 1.00 1.00 1.00 1.00 0.00 Monatin 1.13 0.86 1.44 1.20 1.22 1.17 1.17 0.18 90Mon/l0Reb 0.83 0.84 1.28 1.16 1.09 0.75 0.99 0.21 8OMon/20Reb 1.32 0.97 1.32 1.18 1.12 0.89 1.13 0.18 70Mon/3OReb 1.06 0.84 1.30 1.37 1.15 0.96 1.11 0.20 60Mon/4OReb 0.97 1.00 1.27 1.00 0.89 1.11 1.04 0.13 Monatin without Reb A had a mean linger index value of 1.17. Example 9 [0082] Solutions containing blends of monatin with other steviol glycosides (i.e., stevia leaf extract, rebaudioside-B, rebaudioside-D, stevioside) were exposed to light and underwent similar evaluations as described above for monatin and Reb A blends. Lemon-lime flavored carbonated beverages were exposed to fluorescent light (about 5900 Lux) continuously in a light box at 25 0 C for nine weeks. Solutions of about 9 mM citric acid/trisodium citrate (pH of approximately 3.0) in high purity water were sweetened with mionatin (about 6.4 % SEV) and equimolar steviol glycoside concentrations (about 1.6 % SEV), flavored with lemon-lime flavor, and preserved with approximately 150 ppm sodium benzoate. The solutions were carbonated in four liter batch sizes using an Armfield carbonator with approximately 3.5 liters of carbon dioxide ("C0 2 ") per liter of solution and then transferred to 12 ounce clear, plastic bottles. Caps were placed on the bottles after filling. Multiple bottles of each sample were created since a bottle was sacrificed when the measurements below were taken. [0083] Total indole concentrations (monatin and its intramolecular rearrangement products, monatin lactone and monatin lactam) were quantified by chromatography using an Agilent 1290 Infinity UHPLC at 45*C using with a Waters Acquity UPLC H]SS T3 2.lx]00mm, 1.8 ptm Cis column (Waters Corp., Milford, MA) and UV detection at 280 nm. The compounds were eluted by a gradient mobile phase consisting of (A) 0.1% formic acid in 10 mM ammonium formate solution and (13) 100% acetonitrile. The integrated peak areas for the three (3) compounds were expressed in terms of monatin concentration (mg/L) based on a standard curve prepared for monatin. Monatin lactone and monatin lactan were quantified using the monatin standard curve and empirically determined response factors. 34 WO 2012/083251 PCT/US20111/065643 [0084] Total indole concentrations are shown in Table 25 for each of the blends. The percent of the original indole concentration remaining is shown in parentheses below each value. The addition of RA80, rebaudioside-B ("Reb-B"), rebaudioside-D ("Reb-D"), and stevioside did not reduce the proportion of indole that degraded relative to the solution containing monatin alone (untreated). 35 WO 2012/083251 PCT/US20111/065643 Table 25. Total Indole Concentrations for Various Blends of Monatin and Steviol Glycosides Exposed to Light Steviol Glycoside Blended with Monatin Untreated Reb A RA80 Reb B Reb D Stevioside Steviol Glycoside 0 35 35 32 44 31 (ppm) Steviol Glycoside 0 0.0362 0.0374 0.0398 0.0390 0.0385 (mM)' Total Indole (pp _ 24.6 24.6 24.6 24.5 24.6 24.7 Week 0 (100.0) (100.0) (100.0) (100.0) (100.0) (100.0) 23.8 24.0 24.4 23.3 23.8 19.2 Week 1 (96.6) (97.5) (99.1) (94.8) (96.6) (77.9) 23.7 24.1 25.0 23.4 23.6 23.8 Week 2 (96.3) (97.7) (101.5) (95.3) (96.0) (96.5) 24.7 22.6 23.8 22.0 22.6 23.5 Week 3 (100.3) (91.6) (96.8) (89.4) (91.7) (95.1) 23.2 21.7 23.0 20.9 22.8 23.6 Week 4 (94.3) (87.9) (93.6) (85.3) (92.7) (95.6) 23.7 23.0 22.5 22.4 23.6 23.4 Week 5 (96.6) (93.3) (91.6) (91.2) (96.0) (95.0) 21.7 19.8 21.3 18.8 21.1 20.8 Week 7 (88.3) (80.5) (86.6) (76.5) (85.7) (84.5) 19.4 19.7 20.6 16.6 18.6 19.5 Week 8 (78.7) (79,8) (83.8) (67.8) (75.4) (79.1) 15.2 16.9 15.2 15.4 13.8 15.0 Week 9 (61.8) (68.7) (61.8) (62.9) (56.1) (60.7) 36 WO 2012/083251 PCT/US20111/065643 [0085] The yellow color intensity (b*) of each sample was also measured using a HunterLab ColorQuest XE colorimeter (Hunter Associates Laboratory, Inc., Reston, VA). As shown in Table 26, the addition of RA80, Reb B, Reb D, and stevioside did not reduce yellow discoloration relative to the solution containing monatin alone (untreated). Table 26. Yellow Color Development by Various Blends of Monatin and Steviol Glycosides Exposed to Light Steviol Glycoside Blended with Monatin Untreated Reb A RASO Reb B Reb D Stevioside Steviol Glycoside 0 35 35 32 44 31 (ppm) Steviol Glycoside 0 0.0362 0.0374 0.0398 0.0390 0.0385 (mM) Yellow Color Intensity (b*) Week 0 0.12 0.12 0.14 0.13 0.11 0.12 Week 1 0.18 0.28 0.28 0.25 0.33 0.18 Week 2 0.31 0.27 0.40 0.40 0.44 0.33 Week 3 0.26 0.45 0.37 0.58 0.50 0.35 Week 4 0.43 0.60 0.52 0.76 0.69 0.48 Week 5 0.50 0.59 0.72 0.62 0.64 0.48 Week 7 0.65 0.83 0.76 1.03 0.84 0.81 Week 8 1.25 1.00 1.07 1.66 1.37 1.26 Week 9 1.52 0.94 1.63 1.36 1.84 1.60 [0086] The same samples sacrificed for indole and yellow color measurements above were tasted to evaluate the presence or absence of musty off-flavor (Table 27). The addition of RA80, Reb B, Reb D. and stevioside did not delay the onset of musty off-flavor (all samples were musty after 2 weeks) but the intensity of the musty off-flavored tended to be lower in the presence of steviol glycosides compared to monatin alone (untreated), 37 WO 2012/083251 PCT/US20111/065643 Table 27. Musty Off-Flavor Development by Various Blends of Monatin and Steviol Glycosides Exposed to Light Steviol Glycoside Blended with Monatin Untreated Reb A RA80 Reb B Reb D Stevioside Steviol Glycoside 0 35 35 32 44 31 (ppm) Steviol Glycoside 0 0.0362 0.0374 0.0398 0.0390 0.0385 (mM) "Musty Off-Flavor Intensity Rating Week 0 0 0 0 0 0 0 Week 1 0 0 0 0 0 0 Week 2 3 2 2 3 2 3 Week 3 4 3 3 3 2 3 Week 4 4 4 3 3 2 3 Week 5 4 4 4 3 4 'The intensity of musty off-flavor was rated on a scale from 0 to 9 where 0 = none, 1 = trace, 2 = faint, 3 = slight, 4 = mild, 5 = moderate, 6 = definite, 7 = strong, 8 = very strong, and 9 extreme. Example 10 [0087] Unflavored still beverages (non-carbonated) were exposed to fluorescent light (-5900 Lux) continuously in a light box at 25oC for 10 days. Solutions of about 9 mM citric acid/trisodium citrate (p I of approximately 3.0) in high purity water were sweetened with monatin (about 6.4 % SEV) and equimolar concentrations of steviol glycoside (about 1.6 % SEV) and preserved with approximately 150 ppm sodium benzoate. The solutions were transferred to 60 mL clear, plastic bottles. Caps were placed on the bottles after filling. Multiple bottles of each sample were created since duplicate bottles of each treatment were sacrificed when the measurements below were taken. [0088] Total indole concentrations (monatin and its intramolecular rearrangement products, monatin lactone and monatin lactam) were quantified by reversed phase chromatography using an Agilent 1290 Infinity UHPLC at 45*C with a Waters Acquity UPLC HSS T3 38 WO 2012/083251 PCT/US20111/065643 2.1x1I00nm, 1.8 pm Cis column (Waters Corp., Milford, MA) and UV detection at 280 nm. The compounds were eluted by a gradient mobile phase consisting of (A) 0.1% formic acid in 10 mM ammonium formate solution and (B) 100% acetonitrile. The integrated peak areas for the three (3) compounds were expressed in terms of monatin concentration (mg/L) based on a standard curve prepared for monatin. Monatin lactone and monatin lactam were quantified using the monatin standard curve and empirically determined response factors. [0089] Total indole concentrations are shown in Table 28 for each of the blends. The percent of the original indole concentration remaining is shown in parentheses below each value. The addition of Reb A reduced the proportion of indole that degraded relative to the solution containing monatin alone (untreated), However, addition of RA80, Reb B, Reb D, and stevioside did not reduce the proportion of indole that degraded. Thus, the observed effects for Reb A on indole recovery were unique compared to the other steviol glycosides. 39 WO 2012/083251 PCT/US20111/065643 Table 28. Total Indole Concentrations for Various Blends of Monatin and Steviol Glycosides Exposed to Light Steviol Glycoside Blended with Monatin Untreated Reb A. RA80 Reb B Rub D Stevioside Steviol Glycoside 0 35 35 32 44 31 (ppm) Steviol Glycoside 0 0.0362 0.0374 0.0398 0.0390 0.0385 (mM) Total Indole (pp n) 26.1 25.8 25.8 25.7 25.9 26.2 Day0 (100.0) (100.0) (100.0) (100.0) (100.0) (100.0) 26.2 25.4 25.5 25.1 26.2 25.4 Day 2 (100.4) (98.4) (98,8) (97.7) (101.2) (96.9) 23.7 24.0 22.7 22.6 23.3 22.1 Day 4 (90.8) (93.0) (88.0) (87.9) (90.0) (84.4) 19.4 20.0 18.6 18.7 19.4 19.0 Day 6 (74.3) (77.5) (72.1) (72.8) (74.9) (72.5) 15.8 16.8 13.5 12.5 13.9 13.0 Day 8 (60.5) (65.1) (52.3) (48.6) (53.7) (49.6) 15.7 16.2 14.1 13.3 14.5 13.4 Day 10 (60.2) (62.8) (54.7) (51.8) (56.0) (51.1) [0090] The color of each sample was also measured using a HunterLab ColorQuest XE colorimeter (Hunter Associatcs Laboratory, Inc., Reston, VA). As shown in Table 29, the addition of RA80, Reb B, Reb D, and stevioside did not reduce yellow discoloration relative to the solution containing monatin alone (untreated). However, addition of Reb A reduced yellow discoloration and so its ability to reduce discoloration by monatin solutions exposed to light was unique compared to the other steviol glycosides. 40 WO 2012/083251 PCT/US20111/065643 Table 29. Yellow Color Development by Various Blends of Monatin and Steviol Glycosides Exposed to Light Steviol Glycoside Blended with Monatin Untreated Reb A RA80 Reb B Reb D Stevioside Steviol Glycoside 0 35 35 32 44 31 (ppm) Steviol Glycoside 0 0.0362 0.0374 0.0398 0.0390 0.0385 (mM) Yellow Color Intensity (b*) Day 0 0.02 0.04 0.02 0.03 0.03 0.06 Day 2 0.12 0.11 0.15 0.22 0.16 0.19 Day 4 0.40 0.34 0.64 0.60 0.52 0.71 Day 6 0.53 0.42 0.70 0.68 0.59 0.81 Day 8 1.44 1.23 1.99 2.24 1.90 2.18 Day 10 1.51 1.33 1.83 2.05 1.77 2.06 Example 11 [0091] Whether mixtuI-es of monatin, stevia, and lo han guo, exhibit quantitative sweetness synergy in binary blends with each other was investigated. [0092] Quantitative synergy was identified by evaluating the sweetness response for each of the sweeteners alone, creating concentration-response ("C/R") curves for each sweetener, and then evaluating the sweetness of the various blends against predicted results extrapolated from the C/R curves. [0093] Monatin (2R, 4R-nonatin monopotassium salt) and stevia (95% rebaudioside A) were obtained from Cargill. Lo han gno (PureLo 46% mogroside V) was obtained from Biovittoria. All sweetener samples and sucrose references were prepared in a citric/citrate buffer consisting of citric acid monohydrate (0.158%, w/v) and trisodium citrate dihydrate (0.022%, w/v). 41 WO 2012/083251 PCT/US20111/065643 [0094] Sweetness evaluations were made by matching the sweetness of the test solutions to that of a known sucrose standard. A panel of 8-10 trained, experienced judges was presented with a range (2-12%; w/v) of sucrose reference solutions and a single test sample. Each test solution was monatin, Reb A, lo han guo, or a blend, Panelists were instructed to taste the test sample and then to identify the sucrose reference sample that matched the perceived sweetness of the test sample. If the sweetness of the test sample was judged to fall between that of two references, panelists were instructed to estimate sweetness to one decimal point. A rigorous rinsing procedure was adopted following each sip of test or reference solution to minimize the potential for sweetness adaptation and carry-over. Panelists evaluated each sample in duplicate. Results for each test sample were averaged. [00953 To create the C/R curves, each sweetener was evaluated at 4 or 5 evenly spaced points across a range of sucrose equivalence values ("% SEV"). Monatin was evaluated at 10-40 ppm, stevia at 120-600 ppm, and lo han guo at 200-800 ppm. The sweetener concentrations tested and measured % SEV values are summarized in Table 30. The Beidler equation was applied to these data to create an individual C/R curve for each sweetener. Table 30. Concentration-Response Determinations of Monatin, Reb A, and Lo han guo Sweetener Concentration (ppm) % SEV (% sucrose) Monatin 10 3.9 20 6.2 30 7.8 40 9.1 Stevia 120 3.8 240 5.3 360 5.8 480 6.1 600 7.1 Lo han guo 200 3.9 350 4.6 500 6.0 650 6.9 800 7.0 [0096] Binary blends of the sweeteners were then prepared for the synergy evaluations. The % SEV target for the blends was 7-8% SEV, with varying ratios of sweetness contribution from each sweetener. The predicted % SEV for each blend, extrapolated from the Beidler 42 WO 2012/083251 PCT/US20111/065643 C/R curves prepared from the data set gathered in this specific study from this specific panel, and the actual % SEV measured by sensory panel are reported in Table 31. Table 31. Sucrose Equivalence of Theoretical 8% Sucrose Blends Sweetener Calculated % Sweetener Predicted % Actual % % SEV Blend SEV Ratio Blend SEV SEV Difference (% sucrose Concentration (% sucrose (% sucrose (% sucrose equivalence) (ppm) equivalence) equivalence) equivalence) Stevia/ 1.6/5.8 65/475 7.4 7.4 0 Lo han guo 2.6/5.1 120/355 7.6 7.1 -0.5 3.3/4.3 175/265 7.6 7.2 -0.4 3.9/3.4 240/185 7.3 6.7 -0.6 4.7/2.4 340/115 7.1 6.3 -0.8 Monatin/Stevia 2.2/4.7 5/340 6.9 8.3 1.4 3.2/3.9 8/240 7.1 8.9 1.8 4.1/3.3 11/175 7.4 8.9 1.5 5.1/2.6 15/120 7.7 8.5 0.8 6.0/1.6 19/65 7.6 8.4 0.8 Monatin/ 2.2/5.8 5/475 8.0 8.1 0.1 Lo han guo 3.2/5.1 8/355 8.3 8.0 -0.3 4.1/4.3 11/265 8.4 8.3 -0.1 5.1/3.4 15/185 8.5 8.7 0.2 6.0/2.4 19/115 8.4 7.9 -0.5 [0097] The results demonstrate that stevia/lo han guaO blends are almost additive, as are monatin/ Io han guo blends. However, monatin/stevia blends demonstrated quantitative synergy in that the measured sweetness was much higher than predicted. The percent increased sweetness of the monatin/stevia blends is summarized in Table 32. Table 32. Synergy of Monatin/Stevia Blends Sweetener Concentrations Predicted Measured Sweetness Blend (ppm) % SEV % SEV Synergy (%) Monatin/Stevia 5/340 6.9 8.3 20.3 8/240 7.1 8.9 25.3 11/175 7.4 8.9 20.3 15/120 7.7 8.5 10.4 19/65 7.6 8.4 10.5 43 WO 2012/083251 PCT/US20111/065643 Example 12 [0098] The attributes of monatin and Reb A blends in a tabletop sweetener application were investigated. Ingredients and the lot numbers of ingredients used to make tabletop sweetener formulations are identified in Table 33. Table 33. Ingredients Used Ingredients Lot Number Dextrose* 03186883 R, R, lonatin* 1392 Reb A* 1040 Sugar Retail *Available from Cargill Inc., Minneapolis, MN [0099] Tabletop sweeteners were formulated to be equiswcet to two teaspoons of sucrose (8.5 g). The amount of monatin and Reb A used in each formulation was calculated from the Beidler equations for each sweetener, determined previously in Example 5. The Beidler equation relating sucrose equivalence value ("% SEV") to anhydrous monatin concentration (mg/L) in water was derived as: % SEV =26.7* [monatin] / (69.6 + [imonatin]). For Reb A: % SEV=1 1.26* [Reb A] / (253 F [Reb A]). Dextrose was considered to be 75% as sweet as sucrose in formulating. Packet weight for each sweetener was I gram using dextrose for the majority of the bulk. [00100] Six sweetener blends with varying ratios of monatin and Reb A were evaluated in this study. The blends of monatin and Reb A were expressed on a sweetness basis. Because of monatin's greater sweetness potency it provided 90% of the sweetness in the combination. The other blends ratios were about 100/0, 75/25, 50/50, 25/75 and 0/100 of monatin to Reb A. [00101] To prepare each sweetener sample for tasting, one packet (I grain) of each sweetener blend was added to 170g of reverse osmosis-purified water and stirred to dissolve. Sensory attributes of these products were determined by an expert panel of sixteen applications and sensory scientists from the Cargill Research Center who were all experienced in working with and tasting monatin and Reb A. Samples were evaluated in individual sensory booths. Panelists were offered water and crackers to rinse their palates and then waited one minute between samples. Three samples were evaluated in each tasting 44 WO 2012/083251 PCT/US20111/065643 session. All three tasting sessions were completed in one day. The blind coded samples were presented in a balanced rotation and served at room temperature in 2 fluid ounce souffld cups. [00102] The panelists were asked to evaluate each sample using a nine-point scale. A blind coded sucrose sample was also included in the test and evaluated with the other samples. Six attributes were evaluated, including sweetness intensity and sweetness linger. Mean scores for each of the attributes are compiled in Table 34. Table 34. Average Scores for Various Blends of Monatin and Reb A Compared to Sucrose Ratio of % Sweetness from Monatin/Reb A Attribute 100% 90/10 75/25 50/50 25/75 100% Sucrose Monatin Reb A Control Sweetness 4.2 4.3 4.9 4.4 4.1 4.3 4.6 Onset* Sweetness 5.1 4.9 6.1 5.4 5.9 5.3 4.8 Intensity** Sweetness 3.8 3.9 4.1 4.1 3.2 3.3 4.6 Quality*** Sweetness 6.1 6.2 6.6 6.4 6.4 6.3 5.2 Linger* Bitterness 5.7 5.8 5.9 5.6 6.6 6.5 5.3 Intensity** *9-point onset scale (l="xtremely Slower"; 5-"Same as Control"; 9=" Extremely Faster") **9-point linger scale (1="Extrernely Less"; 5="Same as Control"; 9=" Extremely More") ***9-point quality scale (l="Extrenely Worse"; 5="Sane as Control"; 9=" Extremely Better") [00103] The data show that the sweetness intensity of the 75/25 (6.1), 50/50 (5.4), and 25/75 (5.9) blends was greater than the sweetness intensity of either monatin (5.1) or Reb A (5.3) alone. This result is evidence of the sweetness synergy between the two sweeteners. [00104] The greater sweetness intensity may also have driven the higher overall quality, and sweetness onset scores for the 75/25 (4.1 and 4.9) and 50/50 (4.1 and 4.4) blends relative to either monatin (3.8 and 4.2) or Reb A (3.3 and 4.3) alone. The lower difference from control score for the blends indicated the quality was better and closer to that of the sucrose control. [00105] No substantial differences were observed between treatments with respect to sweetness linger. Though blends of monatin and Reb A typically reduce the lingering sweetness, it is likely that that the higher sweetness intensity of the blends also increased the sweetness linger such that no difference was observed between the blends and either monatin 45 WO 2012/083251 PCT/US20111/065643 or Reb A alone. It was also observed that greater ratios of monatin in the blends reduced the bitterness of Reb A. Example 13 [00106] The objective of this study was to demonstrate sweetness synergy between monatin and Reb A at 4% SEV (Table 35), and monatin and RASO at 7% SEV (Table 36). [00107] A group of 7 to 8 food scientists participated in this study. The participants were all experienced tasters and have been working with monatin and Reb A samples. Sample concentrations (Tables 35 and 36) at 4% and 7% SEV were determined using Beidler equations of each sweetener. Samples were mixed in filtered water, and tasted at room temperature. Each participant received 1.5oz of sample in 2oz souffle cups. They were served a set of two samples at a time, asked to taste the samples, and indicate which sample was sweeter than the other. In each set, one sample was the blend, and the other was either monatin or a Reb A sample. Table 35. Samples at 4% SEV Calculated Using Beidler Curves for Each Sweetener Monatin Reb A Relative Sweetness Contribution (ppm) (ppm) (Monatin/Reb A Blend) 12 0 100/0 0 140 0/100 9 19 80/20 Table 36. Samples at 7% SEV Calculated Using Beidler Curves for Each Sweetener Monatin RA80 (ppm) Relative Sweetness Contribution (ppm) (Monatin/RA80 Blend) 25 0 100/0 0 376 0/100 18 38 80/20 46 WO 2012/083251 PCT/US20111/065643 Table 37. Tasting Results for Monatin and Reb A Samples at 4% SEV % Sample Set Total # of # of Panelists Choosing SEV Panelists the Blend as Sweeter 4% Monatin vs. 7 6 (86%) 80Mon/20RebA Blend 4% Reb A vs. 8 4(50%) 8OMon/2ORebA Blend [00108] As the data in Table 37 show, the 80/20 blend of monatin and Reb A was found to be sweeter compared to monatin alone by 86% of the panelists and compared to Reb A alone by 50% of the panelists at 4% SEV. Table 38. Tasting Results for Monatin and RASO Samples at 7% SEV % Sample Set Total # of # of Panelists Choosing SEV Panelists the Blend as Sweeter 7% Monatin vs. 7 7(100%) 80Mon/20RebA80 7% RA80 vs. 80Mon/20 7 7(100%) RebA80 [00109] As the data in Table 38 show, the 80/20 blend of monatin and RA80 was found to be sweeter compared to both monatin and RASO alone by 100% of the panelists at 7% SEV. [00110] These results demonstrate examples of sweetness synergy between monatin and Reb A samples at different % SEV levels. When the sweeteners blend together the resulting sweetness of the blend is higher than that of the individual sweeteners. Example 14 [00 111] The objective of this study was to assess sweetness linger of no-sugar added applesauce using monatin only compared to variations using blends of monatin with Reb A (95% rebaudioside-A), In this study, monatin (2R, 4R-rnonatin monopotassium salt) and Reb A, both available from Cargill, were combined with commercially-available MOTT'S* 47 WO 2012/083251 PCT/US20111/065643 Natural no-sugar added applesauce. This natural applesauce contained 12 grains of sugar from apples in a /2 cup serving (123 grams). Additional sweetness from monatin only or monatin/Reb A blends in the natural applesauce was about 10% SEV, yielding a similar sweetness intensity as MOTT'S* Original applesauce (25 grams of sugar per 2 serving). Three different sweetened variations of the natural applesauce were prepared (Table 39). Table 39. Formulated Levels of Monatin and Reb A in Commercially Available Natural, No-Sugar Added Applesauce Monatin Reb A Relative Sweetness (ppm) (ppm) Contribution (Monatin/Reb A) 42 0 100/0 30 55 77/23 20 140 56/44 [00112] Sensory evaluation of the sweetened applesauces was conducted by a panel of 6 application and sensory scientists from the Cargill Research Center, who were all experienced in working with and tasting monatin and Reb A. Panelists were provided water and crackers to rinse their palates between tasting samples. Applesauce samples were served at refrigeration temperature (44C) in coded 2 ounce souffle cups. [00113] Two different paired sets of applesauce were evaluated by panelists in a balanced rotation in a single day: (a) 100% nonatin sweetened and 77%/23% monatin/ Reb A sweetened blend and (b) 100% monatin sweetened and 56%/44% monatin/ Reb A sweetened blend, For each set, panelists were instructed to consume the entire sample, cleanse their palate, consume the entire second sample, then select the coded sample which had a longer lingering sweetness. Results are shown in Table 40. 48 WO 2012/083251 PCT/US20111/065643 Table 40. Number (n = 6) of Panelists Indicating Longer Lingering Sweetness of Applesauces Sweetened with Monatin Only Compared to Those with Monatin/ Reb A Blends 77% 56% 100% Monatin/ 100% Monatin/ Monatin 23% Monatin 44% Reb A Reb A 6 0 4 2 [001141 The monatin-sweetened applesauce was identified as having more lingering sweetness by all panelists in comparison to the applesauce sweetened with either the 77%/23% or the 56%/44% blend of monatin to Reb A. [00115] These results indicate that the presence of stevia-based sweeteners, such as Reb A, reduced the lingering sweetness profile of monatin in applesauce. It should also be noted that this effect was observed in the presence of inherent fruit sugars from the apples. Example 15 [00116] The objective of this study was to assess sweetness linger of sweetener blends shown in Table 41 compared to monatin, Reb A, RASO, and sucrose over time. Trained Quantitative Descriptive Analysis ("QDA") panelists evaluated these blends and individual sweeteners mentioned above for sweetness intensity and linger at different sweetness blend ratios of 80/20, 60/40, and 50/50, and at different % SEV levels of 4%, 7%, and 12%. Table 41. Sweetener Blends Sweetener Blend % SEV 1. *80Monatin/20RebA 4%, 7%, and 12% 2. 60Monatin/4ORcbA 7% 3. 5OMonatin/5ORebA 7% 4. 80Monatin/20RA80 7% *In this blend 80% of total sweetness comes from monatin and 20% comes from Reb A; other blends are composed in a similar fashion, 49 WO 2012/083251 PCT/US20111/065643 Trained Panel Method [00117] Training: QDA panelists, highly trained and experienced on-call taste panelists, had two to three sessions of training prior to testing. The panelists were trained with reference sucrose samples to practice rating sweetness intensity on the scale, and to determine best technique for rating sweetness and sweetness linger. They determined the best technique to be to take a sip from the sample, hold sample in the mouth for about 10 seconds to allow the sweetener to reach its peak sweetness, and rate the initial sweetness intensity at peak within 10 seconds. Then they spit out the samples to rate remaining sweetness (linger) in their mouth at every 10 seconds. They waited 10 - 15 minutes between samples and rinsed with sparkling water to help cleanse their palate and prevent carryover from previous samples. All of the sweeteners including the blends were adjusted to match in sweetness intensity initially at 10 seconds evaluation. Sweetness matching was carried out using panelists' feedback during training. [00118] Testing: Panelists were served 1.5 oz of each sample in 2 oz souffid cups at room temperature (68-72*F) for the testing session. They followed the technique determined during training: They took one sip of the sample, held the sample in their mouths for 10 seconds, and rated sweetness intensity at peak. They then spit the sample, and rated remaining sweetness intensity every 10 seconds. The sweetness intensity was rated up to 50 seconds for 4% SEV, up to 70 seconds for 8% SEV, and up to 80 seconds for 12% SEV samples. The panelists evaluated the samples with a 10 - 15 minute break in between samples. They rinsed their mouths with sparkling water after their evaluation was finished. All samples were evaluated in a balanced sequential order one at a time. The panelists were given appropriate sucrose sample (e.g., 4%, 7%, or 12%) to calibrate before the testing session. [00119] Ballot: Samples were rated on a 15 point sweetness intensity scale. none strong 0 1 2 3 4 5 6 7 8 9 10 1] 12 13 14 15 [00120] Data Analysis. Data was collected in replicate, and analyzed by ANOVA at 95% confidence interval using SPSS software (Version 15.0 for Windows). 50 WO 2012/083251 PCT/US20111/065643 [001211 QDA Panel Sample and Panelists Information: Tables 42 to 47 show concentrations (%w/v) of each sample used, and number of panelists attended each test. Samples were prepared in filtered water. Table 42. Concentrations of Monatin, Reb A, 80Monatin/20RebA Blend, and Sucrose Formulated at 7% SEV Name Monatin Reb A Sucrose Concentration Concentration Concentration Sucrose 0% 0% 7% Monatin 0.0036% 0% 0% Reb A 0% 0.0417% 0% 80Mon/20RebA 0.0020% 0.0035% 0% Panelist Number: nine trained panelists Table 43. Concentrations of Monatin, Reb A, 60Monatin/40RebA Blend, and Sucrose Formulated at 7% SEV Name Monatin Reb A Sucrose Concentration Concentration Concentration Sucrose 0% 0% 7% Monatin 0.0036% 0% 0% Reb A 0% 0.0417% 0% 60Monatin/4ORebA 0.0016% 0.0069% 0% Panelist Number: eleven trained panelists Table 44. Concentrations of Monatin, Reb A, SOMonatin/50RebA Blend, and Sucrose Formulated at 7% SEV Name Monatin Reb A Sucrose Concentration Concentration Concentration Sucrose 0% 0% 7% Monatin 0.0036% 0% 0% Reb A 0% 0.0417% 0% 50Monatin/5oRebA 0.0013% 0.0092% 0% Panelist Number: ten trained panelists 51 WO 2012/083251 PCT/US20111/065643 Table 45. Concentrations of Monatin, Reb A, SOMonatin/20RebA Blend, and Sucrose Formulated at 4% SEV Name Monatin Reb A Sucrose Concentration Concentration Concentration Sucrose 0% 0% 4% Monatin 0.0015% 0% 0% Reb A 0% 0.014% 0% 80Monatin/20RebA 0.0003% 0.0084% 0% Panelist Number: eight trained panelists Table 46. Concentrations of Monatin, Reb A, 80Monatin/20RebA Blend, and Sucrose Formulated at 12% SEV Name Monatin Reb A Sucrose Concentration Concentration Concentration Sucrose 0% 0% 12% Monatin 0.0090% 0% 0% 8OMon/2ORebA 0.0050% 0.0062% 0% Panelist Number: nine trained panelists Table 47. Concentrations of Monatin, RA80, 80Monatin/20RASO Blend, and Sucrose Formulated at 7% SEV Name Monatin RASO Sucrose Concentration Concentration Concentration Sucrose 0% 0% 7% Monatin 0.0036% 0% 0% RASO 0% 0.0417% 0% 80Monatin/20RA80 0.0025% 0.0033% 0% Panelist Number: six trained panelists [00122] Resu/ts: Sweetness intensity scores collected over time can be seen in the following tables for monatin, Reb A, and various blends of monatin/RebA (80/20, 60/40, and 50/50) samples formulated at 7%, 4%, and 12% SEV. 52 WO 2012/083251 PCT/US20111/065643 Table 48. Sweetness Intensity Change Over Time for Monatin, Reb A, 80monatin/20RebA, and Sucrose Formulated at 7% SEV Samples Iosec 20sec 30sec 40sec 50sec 60sec 70sec Monatin 7.0 a* 6.3 a 4.5 a 2,9 a 1.6 a 0.6 a 0.1 a Reb A 7.0 a 6.2 a 4.4 a 2.4 ab 1.2 ab 0.4 ab 0.1 a 80Monatin/2ORebA 6.9 a 5.6 b 3.7 b 2.1 be 1.0 be 0.3 be 0.1 a Sucrose 6.9a 5.1 b 3.0 b 1.7 c 0.6 c 0.1 c 0.0 a p-values 0.460 0.011 0.008 0.003 0.001 0.002 0.658 * Means followed by different letters are significantly different from each other at p < 0.05. Table 49. Sweetness Intensity Change Over Time for Monatin, Reb A, 60Monatin/40RebA, and Sucrose Formulated at 7% SEV Samples lOsec 20sec 30sec 40sec 50sec 60sec 70sec Monatin 6.9 a 5.8 a 4.O a 2.7 a 1.8a 0.8 a 0.4 a Reb A 6.7 a 6.1 a 4.2 a 2.8 a 1.6 ab 0.8 a 0.2 a 60Monatin/4ORebA 7.0 a 5.7 a 3.6 a 2.1 b 1.3 b 0.4 b 0.2 a Sucrose 7.0 a 4.6 b 2.7 b 1.7 b 0.6 c 0.1 b 0.1 a p-values 0.201 0.001 0.003 0.025 [0.008 0.006 0.181 Table 50. Sweetness Intensity Change Over Time for Monatin, Reb A, 50Monatin/50RebA, and Sucrose Formulated at 7% SEV Samples 1lOsec 20sec 30sec 40sec 50sec 60sec 70sec Monatin 6.9 a 6.1 a 4.4 a 2.9 a 1.8 a 0.9 a 0.3 a RebA 6.8 a 6.3 a 4.8 a 3.1 a 2.0 a 1.0 a 0.4 a 50Monatin/50RebA 6.9 a 6.0 a 3.8 b 2.3 b 1.1 b 0.4 b 0.0 a Sucrose 6.9 a 4.6 b 2.6 c 1.3 c 0.5 c 0.0 c 0.0 a p-values 0.771 0.000 0.000 0.000 0.000 0.000 0.004 Table 51. Sweetness Intensity Change Over Time for Monatin, Reb A, 8OMonatin/20RebA, and Sucrose Formulated at 4% SEV Samples l0sec 20sec 30sec 4 Osec 50sec Monatin 3.84 a 3.38 a 2.31 a 1.31 a 0.44 a Reb A 3.91 a 3.34 a 2.09 a 1.19 a 0.34 a 80Monatin/20RebA 4.00 a 3.31 a 2.06 a 1.06 ab 0.34 a Sucrose 4.03 a 2.75 b 1.50 b 0.66 b 0.16 a p-values 0.329 0.025 0.014 0.021 0.18 53 WO 2012/083251 PCT/US20111/065643 Table 52. Sweetness Intensity Change Over Time for Monatin, 80Monatin/20RebA, and Sucrose Formulated at 12% SEV Samples lOsec 20sec 30sec 40sec 50sec 60sec 70sec 80sec Monatin 11.9 a 10.6 a 7.9 a 5.5 a 3.6 a 23 a 1.2 a 0.5 a 80Monatin/20RebA* 11.0 a 10.4 a 7.7 a 5.0 b 3.2 a 1.9 b 0,7 b 0.2 b Sucrose 12.0 a 9.2 b 5.8 b 3.3 c 1.7 b 0.5 c 0.0 c 0.0 c p-values 0.382 0.000 0.000 0.000 0.000 0.000 0.000 0.006 *A single Reb A sample was not included in the testing. Based on sweetness dose response characteristics of Reb A, a 12% SEV level with Reb A as a single sweetener cannot be reached. Therefore Reb A was not included here as the sweetness of all samples initially needed to match at 10 seconds of tasting. [00123] Tables 48 to 52 show sweetness intensity scores and the change in the scores over time. Panelists scored sweetness intensity of each sample initially within 10 seconds when the sweetness reached its peak while they were holding the sample in their mouth. Then they spit the samples, and continue evaluating the remaining sweetness (sweetness linger) in their mouth every 10 seconds until panelist no longer received any sweetness, As seen from the tables above, monatin, Reb A, and monatin/Reb A blend (80/20, 60/40, and 50/50) samples scored the same or not statistically significantly different from each other for sweetness intensity at initial 10 seconds of evaluation. In other words initially all of the samples matched in sweetness intensity to each other. Over time a decrease in sweetness intensities were observed for all of the samples. The decrease in sweetness was faster for the monatin and Reb A blend samples compared to either monatin or Reb A samples individually. In other words Monatin/Reb A blends had less sweetness linger compared to the either sweetener alone. For example, in the case of 80Monatin/20RebA blend sample at 7% SEV (Table 48), the sweetness scores for this blend were statistically significantly lower than the sweetness scores for monatin and Reb A samples at 20 and 30 seconds of tasting. Similarly, 60Monatin/40RebA blend (Table 49), and 50Monatin/5ORebA blend (Table 50) samples at 7% SEV scored significantly lower in sweetness than both monatin and Reb A samples at 40, 50, and 60 seconds of tasting. The 80Monatin/20RebA blend samples at 4%, and 12% SEV (Tables 51 and 52) also showed similar results that the sweetness scores of blend samples were lower than the either sweetener alone indicating less sweetness linger over time for the blend samples. These results indicate that different blends (80/20, 60/40, and 50/50) of monatin and Reb A had significantly reduced sweetness linger compared to sweetness linger of monatin and Reb A alone at different % SEV (7%, 4%, and 12%) levels. 54 WO 2012/083251 PCT/US20111/065643 Table 53. Sweetness Intensity Change Over Time for Monatin, RA80, SOMonatin/20RA80, and Sucrose Formulated at 7 % SEV Samples 1Osec 20sec 30sec 40sec 50sec 60sec 70sec Monatin 7.0 a 6.3 a 4.7 a 3.3 a 2.0 a 0.9 a 0.2 a RASO 7.0 a 6.4 a 4.7 a 3.3 a 1.6 b 0.5 b 0.2 a 80Monatin/20RA80 7.0 a 5.8 b 4.1 a 2.6 b 1.0 c 0.1 c 0.0 a Sucrose 6.8 a 4.8 c 2.8 b 1.3 c 0.3 d 0.0 c 0.0 a p-values 0.295 0.003 0.007 0.001 0.001 0.009 0.A58 [00124] Table 53 shows another example of blending monatin with RA80. As the data in Table 53 show, the 8OMonatin/20RA80 blend was significantly lower in sweetness at 20, 40, 50, and 60 seconds of tasting compared to both monatin and RA80 samples indicating less sweetness linger for the blend over time. Example 16 [00125] Methods: A roundtable consensus panel was conducted with 6 expert tasters who are research employees experienced in tasting and evaluating monatin, and Reb D samples. Samples were prepared in filtered water and evaluated for sweetness linger. A category scale of 1 to 9 (1=Not at all lingering, 5=Moderately lingering, 9=Extremely lingering sweetness) was used to measure the sweetness linger. Samples were matched for sweetness at 7% SEV before the evaluation started by the expert tasters. Sample concentrations as well as the lot numbers can be found in Table 54. Tasters were served 1.5 oz of sample in 2 oz souffl6 cups at room temperature (68-72F). They were asked to have a sip of the sample, swirl the sample in their mouths and hold it for 10 seconds. After 10 seconds, the tasters either swallowed or expectorated the sample, and rated the sweetness linger at 30, 60 and 90 seconds. Tasters were asked to cleanse well with water and crackers in between tasting the samples. 55 WO 2012/083251 PCT/US2011/065643 Table 54. Concentrations of Monatin, Reb D(92%), 80Monatin/20RebD Blend, and Sucrose Formulated at 7% SEV Name Lot # Monatin Reb D (92%) Sucrose Concentration Concentration Concentration Sucrose M3WTBO 0% 0% 7% Monatin 25001392 0.0036% 0% 0% Reb D 00018279-1931 0% 0.030% 0% 80Monatin/2ORebD 25001392/ 0.0020% 0.0035% 0% 00018279-1931 Table 55. Sweetness Linger Scores Samples 30sec 60sec 90sec Monatin 6.8 5.0 3.7 Rob D 5.8 4.3 3.0 80Monatin/2OReb D 5.8 4.3 3.2 Sucrose 4.2 2.7 2.2 [00126] Results: As the data in Table 55 show, sweetness linger scores of both 80Monatin/2ORebD blend and Reb D samples were lower than that of the monatin at 30, 60, and 90 seconds of evaluation. In consensus expert tasters indicated that SOMon/2ORebD blend, and Reb D were lingering less in sweetness than monatin, and closer to the sucrose in overall linger. In agreement with the previous trained panel results, this example also showed that blending monatin and Reb D decreased sweetness linger of monatin over time. [00127] It will be apparent to one of ordinary skill in the art from the teachings herein that specific embodiments and examples of the present invention may be directed to one or a combination of the above-indicated aspects, as well as other aspects. 56

Claims (43)

1. What is claimed is: A method of improving a sweetener performance in a food product, the method comprising: adding monatin or a salt thereof to the food product ; and adding steviol glycosides to the food product, wherein the steviol glycosides are added in an amount sufficient such that the food product exhibits at least one of a reduction in discoloration and a reduction in off-flavors after exposure to ultraviolet or visible light, as compared to a comparable food product containing monatin without steviol glycosides.

2. The method of claim 1, wherein the food product comprises between about 10 ppm and about 50 ppm ofrmonatin and between about 10 ppm and about 80 ppm of rebaudioside A.

3. The method of claim I or 2, wherein the food product contains less than about 0.5 ppm of 3-methyl indole after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4,000 lux.

4. The method of any of claims I to 3, wherein the food product retains at least about 65% of its monatin equivalents after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4,000 lux, wherein monatin equivalents are defined as monatin and monatin lactone.

5. The method of any of claims I to 4, wherein the food product does not exhibit any appreciable discoloration or appreciable off-flavors after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4,000 lux.

6. The method of any of claims I to 5, wherein the food product exhibits decreased sweetness linger as compared to a comparable food product containing either monatin or steviol glycosides alone.

7. The method of any of claims I to 6, wherein the food product exhibits greater sweetness than the sweetness expected from sweetness calculations made based on the Bcidler equation.

8. The method of any of claims I to 7. wherein the food product is a beverage. 57 WO 2012/083251 PCT/US20111/065643

9. A method of making a stable beverage, the method comprising: adding monatin or a salt thereof to the beverage; and adding steviol glycosides to the beverage in an amount sufficient such that the beverage retains at least about 65% of its monatin equivalents after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4000 lux, wherein monatin equivalents are defined as monatin and monatin lactone.

10. The method of claim 9, wherein the beverage retains at least about 80% of its monatin equivalents after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4000 lux.

11. The method of claim 9 or claim 10, wherein the beverage is a carbonated beverage.

12. The method of any of claims 9 to 11, wherein the beverage is a still beverage.

13. The method of any of claims 9 to 12, wherein the beverage comprises about 10 ppm to about 80 ppm of rebaudioside A.

14. The method of any of claims 9 to 13, wherein the beverage comprises about 10 ppm to about 50 ppm of monatin,

15. The method of any of claims 9 to 14, wherein the beverage exhibits decreased sweetness linger as compared to a comparable food product containing either monatin steviol glycosides alone.

16. The method of any of claims 9 to 15, wherein the beverage exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation.

17. A beverage comprising: about 10 ppm to about 50 ppm of monatin or a salt thereof; and about 10 ppm to about 80 ppm of steviol glycosides, wherein the beverage retains at least about 65% of its monatin equivalents after 28 days storage at about 22 0 C in the continuous presence of fluorescent lighting of about 4,000 lux, wherein monatin equivalents are defined as monatin and monatin lactone.

18. The beverage of claim 17 wherein the monatin is at least about 90% R,R-monatin. 58 WO 2012/083251 PCT/US20111/065643

19. The beverage of claim 17 or claim 18, wherein the monatin is at least about 95% R,R mronatin.

20. The beverage of any of claims 17 to 19, wherein the beverage retains at least about 80% of its monatin equivalents after 28 days storage at about 22*C in the continuous presence of fluorescent lighting of about 4,000 lux.

21. The beverage of any of claims 17 to 20, wherein the beverage is a carbonated beverage.

22. The beverage of any of claims 17 to 21, wherein the steviol glycosides are at least about 95 wt% rebaudioside A.

23. The beverage of any of claims 17 to 22, wherein the steviol glycosides are chosen from the group consisting of rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, stevioside, dulcoside A, steviolbioside, and a combination thereof

24. The beverage of any of claims 17 to 23, wherein the beverage exhibits decreased sweetness linger as compared to a comparable food product containing monatin without steviol glycosides.

25. The beverage of any of claims 17 to 24, wherein the beverage exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation.

26. A beverage composition comprising about 10 ppm to about 50 ppm of monatin or a salt thereof; and about 10 ppm to about 80 ppm of steviol glycosides, wherein the beverage composition does not exhibit any appreciable odors or any appreciable off-flavors after 28 days storage at 22 'C in the continuous presence of fluorescent lighting of about 4,000 lux.

27. The beverage composition of claim 26, wherein the beverage composition is a carbonated beverage.

28. The beverage composition of claim 26 or claim 27, wherein the monatin comprises at least about 90% R,R-monatin. 59 WO 2012/083251 PCT/US20111/065643

29. The beverage composition of any of claims 26 to 28, wherein the steviol glycosides include rebaudioside A.

30. The beverage composition of any of claims 26 to 29, wherein the steviol glycosides are at least about 80 wt% rebaudioside A.

31. The beverage composition of any of claims 26 to 30, wherein the steviol glycosides are at least about 95 wt% rebaudioside A.

32. The beverage composition of any of claims 26 to 31, wherein the steviol glycosides are chosen from the group consisting of rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, stevioside, dulcoside A, steviolbioside, and a combination thereof.

33. The beverage of any of claims 26 to 32, wherein the beverage exhibits decreased sweetness linger as compared to a comparable food product containing monatin without steviol glycosides.

34. The beverage of any of claims 26 to 33, wherein the beverage exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation.

35. A beverage composition comprising about 10 ppm to about 50 ppm of monatin or a salt thereof; and about 10 ppm to about 80 ppm of steviol glycosides, wherein the beverage does not exhibit any appreciable discoloration after 28 days storage at 22 *C in the continuous presence of fluorescent lighting of about 4,000 lux.

36. The beverage composition of claim 35, wherein the beverage composition is a carbonated beverage,

37. The beverage composition of claim 35 or claim 36, wherein the monatin comprises at least about 90% R,R-monatin.

38. The beverage composition of any of claims 35 to 37, wherein the steviol glycocides include rebaudioside A.

39. The beverage composition of any of claims 35 to 38, wherein the steviol glycosides are at least about 80 wt% rebaudioside A. 60 WO 2012/083251 PCT/US20111/065643

40. The beverage composition of any of claims 35 to 39, wherein the steviol glycosides are at least about 95 wt% rebaudioside A.

41. The beverage composition of any of claims 35 to 40, wherein the steviol glycosides are chosen from the group consisting of rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, stevioside, dulcoside A, steviolbioside, and a combination thereof.

42. The beverage of any of claims 35 to 41, wherein the beverage exhibits decreased sweetness linger as compared to a comparable food product containing monatin without steviol glycosides.

43. The beverage of any of claims 35 to 42, wherein the beverage exhibits greater sweetness than the sweetness expected from sweetness calculations made using the Beidler equation. 61