Flavor and flavor intensifying composition comprising glycosylate steviol glycoside, food product or beverage with an intense flavor and flavor profile and method for increasing flavor and flavor intensity of a food product or beverage

Description

1 / 34 “FLAVOR AND TASTE ENHANCING COMPOSITION COMPRISING GLYCOSYLATED STEVIOL GLYCOSIDE, FOOD OR BEVERAGE PRODUCT WITH AN INTENSE FLAVOR AND TASTE PROFILE AND METHOD FOR INCREASING THE INTENSITY OF FLAVOR AND TASTE OF A PRODUCT OF FOOD OR DRINK Background of the Invention

[001] Divided from BR 11 2013 023964 6, deposited on 03 / 22 / 2012.

[002] Stevia rebaudiana Bertoni is a perennial shrub of the Asteraceae (Compositae) family that is native to certain regions of South America. The leaves of the plant contain 10 to 20% diterpene glycosides, which are about 150–450 times sweeter than sugar. The leaves have been traditionally used for hundreds of years in Paraguay and Brazil to sweeten teas and local medicines.

[003] Currently, there are over 230 species of Stevia with significant sweetening properties. The plant has been successfully cultivated in a wide range of conditions from its native subtropical area to the cold northern latitudes.

[004] Stevia rebaudiana plant extract contains a mixture of different sweet diterpene glycosides, which have a single base – steviol – and differ by the presence of carbohydrate residues at positions C13 and C19. These glycosides accumulate in stevia leaves and make up about 10%–20% of the total dry weight. Typically, based on dry weight, the four main glycosides found in stevia leaves are Dulcoside A, Rebaudioside C, Rebaudioside A, and Stevia. Other glycosides identified in stevia extract include Rebaudioside B, D, E, and F, Steviolbioside, and Rubusoside.

[005] Steviol glycosides differ from one another in sweetness as well as other sensory characteristics that contribute to taste quality, such as bitterness, long aftertaste, etc. Kinghorn, AD, Stevia: The genus Stevia, Taylor & Francis, London (2002). The taste quality of a glycoside Petition 870260041436, dated 04 / 05 / 2026, p. 10 / 51 The 2 / 34 steviol content is one of the key characteristics that is decisive for its use in various food and beverage applications. The sweetness and taste qualities of stevia glycosides are shown below in Table 1. Table 1: Steviol glycoside molecules and their sweetness and taste quality Steviol Glycosides R1 (C-19) R2(C-13) Sweetening Power relative to Sucrose Taste Quality Stevioside (G3) β-Glc β-Glc -β-Glc (2-1) 150-300 0 Rebaudioside A (G4) β-Glc β-Glc -β-Glc (2η-β-Glc (3-1) 200-400 +2 Rebaudioside B (G3) H β-Glc -β-Glc (2-1)-β-Glc (3-1) 300-350a 0 Rebaudioside C (G4) β-Glc β-Glc -β-Rha (2- 1)-β -Glc (31) 50-120 -1 Rebaudioside D (G5) β-Glc -β -Glc (2-1) β-Glc -β -Glc (2 1)-β -Glc (3-1) 200 - 300 +3 Rebaudioside E (G4) β-Glc-β -Glc (2-1) β-Glc -β -Glc (2 - 1) 250 - 300 + 1 Rebaudioside F (G4) β-Glc β-Glc -β -Xyl (2 1)-β -Glc (3-1) Na Na Rubusoside (G2) β-Glc β-Glc 110 -2 Steviolmonoside (G1) H β-Glc Na Na Steviolbioside (G2) H β-Glc -β -Glc (2 - 1) 100-125 -3 Dulcoside A (G3) β-Glc β-Glc -a -Rha (2 -1) 50-120 -2

[006] Previous studies show a certain correlation between the number of glycosidic residues and the taste quality of a steviol glycoside. When comparing steviol glycosides, rebaudioside A (G4, with four residues of Petition 870260041436, dated 04 / 05 / 2026, page 11 / 51 3 / 34 glucose) clearly surpasses stevioside and rebaudioside B (G3, each with 3 glucose residues) in taste quality. Steviolbioside and rubusoside (G2, each with two glucose residues) have a taste quality that is proven to be significantly inferior to that of steviosides (G3). Furthermore, the taste quality of rhamnosylated glycosides is inferior compared to that of glycosylated glycosides. Tanaka, O., Improvement of Taste of Natural Sweeteners, Pure & Appl. Chem,. Vol. 69, No. 4, pp 675 - 683 (1997).

[007] The sweetness and taste quality of steviol glycoside molecules containing only glucose units are represented in Figures IA - IB. Steviol glycosides with a greater number of glucose residues, for example, more than two glucose residues, show a better taste quality. In particular, the mono- and di-glycosyl forms of stevioside (with 4 glucose residues (G4) and 5 glucose residues (G5), respectively) have significantly better taste quality. Tanaka, O., “Improvement of Taste of Natural Sweeteners” Pure & Appl. Chem., Vol. 69, No. 4, pp. 675 - 683 (1997).

[008] It is known that undesirable flavor attributes can be substantially reduced or eliminated through the intermolecular transglycosylation reaction of various enzymes, in which the attachment of new carbohydrates occurs at the C13 and C19 positions of steviol glycosides. Tanaka (1997) evaluated the effect of adding glucose molecules to stevioside molecules purified by transglycosylation. The resulting glycosylated steviosides were evaluated for their sweetness and taste quality. The improvement in taste quality was greater when glucose units were added at the C19 position than at the C13 position.

[009] Several enzymes have been used to conduct such transglycosylation. Pullulanase, isomaltase (Lobov, SV et al., “Enzymic Production of Sweet Stevioside Derivatives: Transglucosylation by Glucosidases” Agric. Biol. Chem., Vol. 55, No. 12, pp. 2959 - 2965 10 (1991), β-galactosidase (Kitahata, S. et al., “Production of Petition 870260041436, dated 04 / 05 / 2026, page 12 / 51 4 / 34 Rubusoside Derivatives by Transgalactosylation of Various (3-Galactosidases,” Agric. Biol. Chem., Vol. 53, No. 11, pp. 2923-2928 (1989)), and dextrin sucrase (Yamamoto, K. et al., Biosci. Biotech. Biochem., Vol. 58, No. 9, pp. 1657-1661 (1994)) were used as enzymes with pullulan, maltose, lactose, and partially hydrolyzed starch as donors.

[0010] Transglycosylation of steviol glycosides was also achieved by the action of cyclodextrin glucanotransferase (CGTase). The sweeteners obtained had improved sweetness without the bitter, licorice taste (US Patent Nos. 4,219,571, 7,838,044 and 7,807,206).

[0011] With an increase in the number of glucose units in steviol glycoside molecules (e.g., from stevioside to rebaudioside A), the sweetness intensity increases and the sweetness profile (taste) improves. However, the relative sweetness does not increase significantly beyond a certain level even with a further increase in glucose units, as shown in Figure 1a. Published data show that the quality of sweetness improves with the addition of glucose units, but it does not explicitly or implicitly mention that the addition of glucose units contributes to a reduction in sweetness. Brief Summary of the Invention

[0012] The present invention relates to a composition profile that enhances the taste and flavor profile. The composition includes glycosylated steviol glycosides that can increase the intensity of a taste and / or flavor in a food or beverage product. In some embodiments, the glycosylated steviol glycosides may include a plurality of glucose units. For example, the glycosylated steviol glycosides may include three, four, five, or more than five glucose units.

[0013] The present invention also relates to a food product. Petition 870260041436, dated 04 / 05 / 2026, page 13 / 51 5 / 34 or beverage with an intense flavor and taste profile, wherein the food or beverage product includes a flavor and taste-enhancing composition comprising glycosylated steviol glycosides. A wide range of food and beverage products, such as, but not limited to, soft drinks, fruit juices, dairy products, milk drinks, bakery / baked goods, cereal-based products and tabletop sweeteners, can be made in accordance with the present invention. The flavor and taste profile of a food or beverage product including a flavor and taste-enhancing composition, wherein the flavor and taste-enhancing composition includes glycosylated steviol glycosides, can be more intense than a comparative flavor and taste profile of a comparative food or beverage product that does not include the flavor and taste-enhancing composition.Furthermore, the taste of a beverage or food including the flavor-enhancing composition, where the flavor-enhancing composition includes glycosylated steviol glycosides, may be improved relative to the taste of a comparative food or beverage product that does not include the flavor-enhancing composition.

[0014] The present invention further relates to a method for increasing the intensity of taste and flavor of a food or beverage product, including the step of adding a taste and flavor enhancing composition to the food or beverage product, wherein the taste and flavor enhancing composition includes glycosylated steviol glycosides. The present invention also relates to a method of improving the organoleptic properties of a food or beverage product, including a high-fructose syrup, including the step of adding a taste and flavor enhancing composition to the food or beverage product. For example, adding the taste and flavor enhancing composition can cause the high-fructose syrup, such as high-fructose corn syrup, to taste more like sugar. Furthermore, if the fructose syrup is high-fructose corn syrup 42 (HFCS 42), the addition of the enhancing composition Petition 870260041436, dated 04 / 05 / 2026, p. 14 / 51 6 / 34 of flavor and taste may cause HFCS 42 to taste more like high fructose corn syrup 55 (HFCS 55).

[0015] The present invention also relates to a method of producing the flavor and taste-enhancing composition which includes the steps of: extracting steviol glycosides from the leaves of a Stevia rebaudiana Bertoni plant; and transglycosylating steviol glycosides to add glucose units to the steviol glycosides. In some embodiments, the transglycosylation of steviol glycosides includes enzymatic transglycosylation using an enzyme. Examples of enzymes that can be used according to the present invention include, but are not limited to, pullulanase, isomaltase, β-galactosidase, dextrin sucrase, and cyclodextrin glucotransferase.

[0016] The present invention further relates to a method of manufacturing a food or beverage product, which includes the steps of: adding the flavor and taste-enhancing composition, including glycosylated steviol glycosides, and adding a reduced amount of erythritol, where the reduced amount of erythritol is less than an amount of erythritol found in a comparative composition of a food or beverage product that does not include the flavor and taste-enhancing composition. The taste of the food or beverage product is similar to the taste of the comparative food or beverage product, despite the comparative food or beverage product containing high levels of erythritol.

[0017] The above description has highlighted quite broadly the characteristics and technical advantages of the present invention so that the following description of the details of the invention may be better understood. The additional features of the invention, which form the subject matter of the invention claims, will be described below. It should be understood by those skilled in the art that the specific embodiments disclosed can be readily used as a basis for modifying or designing other forms or structures for the Petition 870260041436, dated 04 / 05 / 2026, page 15 / 51 7 / 34 execution of the same purposes as the present invention. It should also be understood by those skilled in the art that such equivalent constructions do not depart from the spirit and scope of the invention, as defined in the appended claims. The novel features, which are believed to be characteristic of the invention, both in terms of their organization and method of operation, together with other objects and advantages, will be better understood from the following description. Brief Description of the Figures

[0018] FIG. 1a represents a bar graph showing the relative sweetness of steviol glycoside molecules in relation to increasing glucose units. FIG. 1b is a bar graph showing the taste quality of steviol glycoside molecules in relation to increasing glucose units.

[0019] FIG. 2 shows the chemical structures of stevioside and glycosyl stevioside.

[0020] FIG. 3 is a graph of the sweetness potency, or equivalent sweetness value (SEV), of stevia glycosides at a sugar sweetness level of 5%. Thus, the aforementioned graph refers to the effect of glycosylation on sweetness.

[0021] FIG. 4a is a graphical representation of the relative classifications of sweetness intensity and the appearance of sweetness in mango and passion fruit flavored beverages to which different stevia ingredients were added, among which sugar was not included. Thus, the aforementioned graphical representation refers to the effect of glycosylation on the sweetness of the beverage (without added sugar). FIG. 4b is a graph of the relative classifications of passion fruit and mango flavor in mango and passion fruit flavored beverages. Thus, the aforementioned graphical representation refers to the effect of glycosylation on Petition 870260041436, dated 04 / 05 / 2026, p. 16 / 51 8 / 34 Modification of beverage flavor (without sugar).

[0022] FIG. 5a is a graphical representation of the relative classifications of sweetness intensity and the appearance of sweetness in mango and passion fruit flavored beverages to which different stevia ingredients, including sugar, were added. Thus, the aforementioned graphical representation refers to the effect of glycosylation on the sweetness of the beverage (with sugar). FIG. 5b is a graph of the relative classifications of passion fruit and mango flavor in mango and passion fruit flavored beverages. Thus, the aforementioned graphical representation refers to the effect of glycosylation on the modification of the beverage's flavor (with sugar).

[0023] FIG. 6 is a graph of the contribution of the SEV (equivalent sweetness value) of NSF-O2 as a function of the concentration of NSF-O2 in ppm, in acidified water and in acidified sugar solutions. Thus, the aforementioned graphical representation refers to the synergy of NSF-O2 in the acidified solution. NSF-O2 (Natural Sweet Flavor #2) contains glycosylated steviol glycosides and dextan, and is discussed further below.

[0024] FIG. 7 is a graph showing the synergy of NSF-O2 with stevia extract, in a batch with reduced stevia concentration versus NSF-O2 concentration. This graphical representation refers to the synergy of NSF-O2 with stevia extract. Detailed Description of the Invention

[0025] It has been unexpectedly discovered that glycosylation of steviol glycosides above a certain number of glucose units efficiently reduces sweetness. It has also been discovered that, with the reduction in sweetness, glycosylated steviol glycosides can contribute to flavor modification and sweetness profile. Therefore, while sweetness decreases with glycosylation, flavor alteration increases. The mixture of steviol glycosides provides a certain amount of sweetness, but the present invention shows that glycosylated steviol glycosides Petition 870260041436, dated 04 / 05 / 2026, page 17 / 51 9 / 34 (hereinafter GSG) improves flavor and sweetness profile in a wide range of applications, such as, but not limited to, the categories shown below in the Table. 2. Table 2: GSG Applications Application Product Results Beverages Apple and Blueberry Juice Significantly higher intensity of blueberry flavor compared to the control. Directionally, more acceptable in terms of overall flavor. Orange, Pineapple, and Passion Fruit Carbonated Beverages Significantly higher intensity of orange flavor, pineapple flavor, and sweetness. Bakery Products Lemon Poppy Seed Muffins Significantly higher intensity of total flavor and sweetness compared to the control. Directionally, the test sample was more acceptable overall and had a higher intensity of lemon flavor. Processed Fruits Strawberry Syrup Significantly less bitter compared to the control Dairy Products Banana Flavored Milk Beverage Significantly higher intensity of total flavor, banana flavor, sweetness, acidity, and bitterness.

[0026] Similar improvements in flavor and taste have been found in other product categories, including, but not limited to, tabletop sweeteners, sauces, confectionery products, baked goods, cereals, snacks, and fruit and vegetable preparations. Examples

[0027] In the following examples, the percentages in the formulas refer to percentages by weight. Example 1: Evaluation of the Iso-sweetness of Steviol Glycosides Petition 870260041436, dated 04 / 05 / 2026, p. 18 / 51 10 / 34

[0028] To evaluate the iso-sweetness of steviol glycosides (SG) and glycosylated steviol glycosides (GSG), a series of samples were selected, as shown below in Table 3. The GSGs were produced by treating the raw materials, steviol glycosides extracted from the Stevia plant, and starch extracted from tapioca with a natural enzyme. The enzyme transfers glucose units from the starch to the steviol glycosides. The enzyme used to facilitate this transfer is produced by fermentation using non-GMO (genetically modified organism) bacteria.

[0029] FIG. 2 is an illustration of an example of glycosylation. Specifically, FIG. 2 illustrates the single glycosylation (G1) of a stevioside molecule. This process can result in multiple glycosylations (G2, G3, etc.) of different steviol glycosides (mainly stevioside and rebaudioside A) present in stevia extract. Table 3 Ingredients of the Test Molecular Weight (med.) Steviol Equivalent Non-Glycosylated Steviol Glycosides Glycosylated Steviol Glycosides G1 & G2 G3-G9 G10-G20 Stevioside 805 0.396 > 99% Reb A 967 0.329 > 99% Reb D 1129 0.282 > 99% GSG-S 1210 0.263 < 10% 95% GSG-M 1380 0.231 7.80% 42% 50% GSG-L 1798 0.177 5% 19% 60% 16%

[0030] To evaluate the sweetness potency of various concentrations of stevia products in aqueous solutions, aqueous solutions of sugar, stevioside, rebaudioside A (Reb A), rebaudioside D (Reb D), GSG-S (contains mainly smaller GSGS, with 1 or 2 glucose units added to SG), GSG-M and GSG-L at various concentrations were prepared using bottled water. Samples were evaluated by assessors at room temperature 21.1 - 22.2 °C (70 - 72 °F). Petition 870260041436, dated 04 / 05 / 2026, p. 19 / 51 11 / 34

[0031] The evaluators were participants previously qualified for their acuity of taste and trained in the use of a sweetness intensity rating scale. The evaluations were performed in duplicate using the same panel participants so that a total of 22 values ​​were generated for each mean data point. Before conducting the study, the evaluators were trained with sugar solutions and the use of the ballot.

[0032] Samples were given sequentially to the evaluators and coded with three-digit numbers. The order of sample presentation was randomized to avoid bias in the order of presentation. A five-minute rest period was provided between samples. Water and unsalted biscuits were provided to cleanse the palate.

[0033] The results were statistically analyzed to generate a standard error value for each of the solutions, as well as a 95% confidence level. The sweetness potency of the different stevia ingredients was estimated by comparing the sweetness of each ingredient in the test with the sweetness of the various sucrose solutions, as shown in FIG. 3. FIG. 3 is a graph of the sweetness potency, or equivalent sweetness value (SEV), of the different stevia ingredients at a 5% sugar sweetness level (i.e., at a concentration corresponding to 5% sucrose). This figure shows the effect of glycosylation on the SEV of steviol glycosides. As the number of glucose units in the steviol glycosides increases, the sweetness of the stevioside also increases for Reb A and then begins to decrease with additional glucose units. Example 2: Effect of Glycosylation on Flavor Modification

[0034] A mango-passion fruit flavored water formula was developed to evaluate the effect of different stevia ingredients on the sweetness and flavor profile of the beverage. A total of 9-10 panel members participated in this sensory test, where they assigned values ​​related to sweetness, appearance Petition 870260041436, dated 04 / 05 / 2026, p. 20 / 51 12 / 34 of the sweetness, to the Mango flavor, to the Passion Fruit flavor, to the acidity, to the overall flavor, etc. Example 2A: Effect of Glycosylation on the Flavor Modification of Sugar-Free Beverages

[0035] Table 4 shows the formula for sugar-free beverages that primarily used Reb A, Reb D, GSG-S, or GSG-L. The quantity of each ingredient (Reb A: 150 ppm; Reb D: 165 ppm; GSG-S: 190 ppm, and GSG-L: ​​300 ppm) was chosen to have approximately 50 ppm of steviol in each formula. Table 4: Mango and Passion Fruit Flavored Beverages Without Added Sugar Sugar Stevia Ingredients Reb A Reb D GSG-S GSG-L Water 95.82 95.82 95.82 95.82 SG Content 0.015 0.0165 0.019 0.03 Citric Acid 0.078 0.078 0.078 0.078 Sodium Citrate 0.056 0.056 0.056 0.056 Mango Flavor 0.031 0.031 0.031 0.031 Passion Fruit Flavor 0.014 0.014 0.014 0.014

[0036] FIGS. 4A - 4B show the modification of flavor and sweetness profiles caused by glycosylation. The intensity of sweetness decreased and the appearance of sweetness was delayed with glycosylation. The Mango flavor was enhanced and the Passion Fruit flavor was reduced with glycosylation. Example 2B: Effect of Glycosylation on the Flavor Modification of a Sugary Beverage

[0037] Table 5 shows the same formula for the Mango and Passion Fruit flavored drinks with 4% sugar and stevia ingredients that contribute an additional 4% sugar-equivalent sweetness. The formula used Reb A, Reb D, GSG-S, or GSG-L in amounts of 50, 55, 73, and 200 ppm, respectively, as described in the formula below. As the number of glucose units increased, greater modification in flavor (either by increase or suppression) was observed as shown in FIGS. 5A - 5B. Petition 870260041436, dated 04 / 05 / 2026, p. 21 / 51 13 / 34 Table 5: Mango and Passion Fruit Flavored Beverages (With Added Addition of Sugar) Stevia Ingredients Reb A Reb D GSG-S GSG-L Water 95.82 95.82 95.82 95.82 Sugar 4.00 4.00 4.00 4.00 SG Content 0.005 0.0055 0.0073 0.02 Citric Acid 0.078 0.078 0.078 0.078 Sodium Citrate 0.056 0.056 0.056 0.056 Mango Flavor 0.031 0.031 0.031 0.031 Passion Fruit Flavor 0.014 0.014 0.014 0.014

[0038] FIGS. 5A - 5B show the effect of SG glycosylation on modifying the flavor and sweetness profiles of a beverage sweetened with sugar and stevia (SG). The sweetness intensity decreased and the onset of sweetness was delayed with glycosylation. Both Mango and Passion Fruit flavors were somewhat suppressed with glycosylation. A key point is that GSG modified the flavor profile. Example 3: Soft drinks with Glycosyl Steviol Glycosides

[0039] In this and the following examples, GSG can be any glycosyl steviol glycoside composition, such as, but not limited to, a combination of GSG-S, GSG-M, and GSG-L. To evaluate changes in the flavor of a soft drink, a range of GSG concentrations (0 to 1,000 ppm) was used with a typical Apple and Blueberry soft drink formula. The objective was to evaluate whether the addition of GSG has an effect on the main flavor attributes in various beverage applications. Specifically, the objective was to determine whether the flavor profile and overall acceptance of a control sample with Apple and Blueberry juice (containing no GSG) differs by 30% from a test sample of the same beverage (containing GSG). After preliminary sensory tests, it was found that GSG modified the sweetness and flavor profile at all concentrations. The GSG concentration is preferably in the range of approximately 0 to 1.000 ppm, more preferably in the range of about 50 to 750 ppm, and, ideally, in the range of about 50 to 500 ppm. Petition 870260041436, dated 04 / 05 / 2026, page 22 / 51 14 / 34

[0040] For the detailed sensory tests, two samples were selected to test against the control sample. The methodology for the detailed sensory tests is indicated below in Table 6. The formulas for the control and test samples are indicated below in Table 7. Table 6: Methodology • Number of sessions: 1 • Number of participants: 20 • Test design: Balanced, randomized within pairs. Blinded • Sensory test method: Intensity and acceptance ratings • Environmental condition: Standard booth lighting • Attributes and scales: • Overall acceptance on a 6-point hedonic scale, where 5 = Extremely liked, 2 = Neither liked nor disliked, and 0 = Disliked • Overall flavor, Apple flavor, Berry flavor, Sweetness, Acidity, Bitterness, and intensity of prolonged residual sweet taste on a 6-point continuous intensity scale, where 0 = Imperceptible and 5 = Extremely pronounced • General Open-ended Comments • Consumption temperature: Refrigerated (~45°F or 7.22°C) • Instructions on consumption / by participants: Samples were served simultaneously. Participants were instructed to read the ingredient list and rate each sample. Table 7: Samples Flavor Description Control Test-1 Low GSG Levels Test-2 High GSG Levels Apple Juice Concentrate 6.21% 6.21% 6.21% Blueberry Juice Concentrate 0.76% 0.76% 0.76% Apple and Blueberry Flavor 0.12% 0.12% 0.12% Sugar 5.60% 5.60% 5.60% GSG 0.025% 0.05% Water Balance Balance Balance

[0041] In this study, twenty members of the consumer panel evaluated Petition 870260041436, dated 04 / 05 / 2026, page 23 / 51 15 / 34 Three samples of flavored apple and blueberry juice were evaluated for overall acceptability and intensity of apple and blueberry flavors, flavor appearance, sweetness, and aftertaste (including acidity, bitterness, and intensity of prolonged residual sweetness). The three samples included a control sample with no glycosyl steviol glycosides (GSG) and two test samples containing low (0.025%) and high (0.05%) levels of GSG. The aim of the test was to determine if the addition of glycosyl steviol glycosides affects the flavor profile of a juice. The results indicated that: - The test samples had greater overall acceptance and a more intense apple flavor than the control sample (with > 90% confidence). - The sweetness intensity of the test sample with low levels of GSG was not significantly different from the control. The use of high levels of GSG increases sweetness and flavor (p = 0.047). - There was no significant difference in the intensity of the aftertaste between the test and control samples (with 90% confidence). Example 4: Carbonated Soft Drink Flavored with Glycosyl Steviol Glycosides

[0042] To evaluate the flavor modification of a carbonated beverage, a range of GSG concentrations (0 to 1000 ppm) was used in an orange-pineapple flavored soft drink formula. The objective was to evaluate whether the addition of GSG has an effect on the main flavor attributes in various carbonated soft drink (CSD) applications. Specifically, the objective was to determine if the flavor profile and overall acceptance of a control sample of orange-pineapple flavored carbonated beverage differs from test samples of the same beverage containing GSG. After preliminary sensory tests, it was found that GSG modified the flavor and sweetness profile at all concentrations. The GSG concentration is preferably in the range of Petition 870260041436, dated 04 / 05 / 2026, page 24 / 51 16 / 34 approximately 0 to 1,000 ppm, more preferably in the range of approximately 25 to 750 ppm, and, primarily, in the range of approximately 50 to 500 ppm.

[0043] For the detailed sensory tests, two samples were selected to test against the control sample. The methodology for the detailed sensory tests is indicated below in Table 8. The formulas for the control and test samples are indicated below in Table 9. Table 8: Methodology • Number of sessions: 1 • Number of participants: 24 • Test design: Balanced, randomized within pairs. Blinded. • Sensory test method: Intensity and acceptance ratings • Environmental condition: Standard booth lighting • Attributes and scales: • Overall acceptance on a 6-point hedonic scale, where 5 = Liked and 0 = Disliked. Overall flavor, Orange flavor, Pineapple flavor, Long-lasting taste, Sweetness on a 6-point continuous intensity scale, where 0 = Imperceptible and 5 = Extremely pronounced. • General Open-ended Comments • Statistical Analysis: ANOVA (by block) with Duncan's post-hoc test. Sample size: ~44.26 mL (1.5 oz) in a clear, capped plastic cup. • Consumption temperature: Refrigerated temperature (~45°F or 7.22°C). • Instructions on consumption / by participants: Samples were served simultaneously. Participants were instructed to read the list of ingredients and evaluate each sample. Table 9: Samples Ingredients Control (%) Test 1 (%) Test 2 (%) Water 90.8 90.8 90.8 Sugar 8.93 8.93 8.93 Petition 870260041436, dated 04 / 05 / 2026, page 25 / 51 17 / 34 Acids 0.142 0.142 0.142 Orange Flavor 0.069 0.069 0.069 Pineapple Flavor 0.050 0.050 0.050 SG95 0.006 0.006 0.006 GSG 0.025 0.05

[0044] In this study, twenty-four consumer panel members evaluated three samples of orange- and pineapple-flavored carbonated beverages for overall acceptability and the intensities of the attributes (overall flavor, orange flavor, pineapple flavor, sweetness, and aftertaste). The three samples included: a reduced-sugar control sample containing SG95 (a stevia extract) and two test samples, which are the same as the control samples plus the addition of low (0.025%) and high (0.05%) levels of SG95. SG95 is a high-purity stevia sweetener available from PureCircle, 915 Harger Road, suite 250, Oak Brook, Illinois 60523, USA. The objective of the test was to determine whether the addition of solid stevia extracts affects the flavor profile of a reduced-sugar carbonated beverage. The results indicated that: - The test sample with low GSG levels had a much more intense orange flavor than the control sample (95% confidence). The test sample with high GSG levels also had a more intense orange flavor than the control sample (p = 0.089).

[0045] There was no significant difference in overall acceptance, pineapple flavor intensity, or aftertaste intensity between the control and the two test samples (with 90% confidence). Example 5: Flavored Milk Beverage with Glycosyl Steviol Glycosides

[0046] To evaluate changes in the flavor of a flavored dairy beverage, a range of GSG concentrations (0 to 1,000 ppm) was used in a banana-flavored flavored dairy beverage formula. The objective was Petition 870260041436, dated 04 / 05 / 2026, page 26 / 51 18 / 34 To determine whether the addition of glycosyl steviol glycosides (GSG) has an effect on key flavor attributes and / or enhances flavor perception in various beverage applications, specifically in dairy beverages. Specifically, the objective was to determine whether the flavor profile and overall acceptance of a control sample of banana-flavored dairy beverage (containing no GSG) differs from two test samples of the same beverage containing two levels of GSG. Although the test was conducted with banana flavor, the results are also relevant for all fruit, vegetable, chocolate, coconut-flavored beverages and energy drinks. After preliminary sensory testing, it was found that GSG modified the sweetness profile and flavor at all concentrations. The GSG concentration is preferably in the range of about 0 to 1,000 ppm, more preferably in the range of about 25 to 750 ppm, and, ideally, in the range of about 50 to 500 ppm.

[0047] For the detailed sensory tests, two samples were selected to test against the control sample. The methodology for the detailed sensory tests is indicated below in Table 10. The formulas for the control and test samples are indicated below in Table 11. Table 10: Methodology • Number of sessions: 1 • Number of participants: 32 • Test design: Balanced, randomized within the pair. Blinded. • Sensory test method: Intensity and acceptance ratings • Environmental condition: Standard booth lighting • Attributes and scales: Overall acceptance on a 9-point hedonic scale, where 9 = Extremely liked, 5 = Neither liked nor disliked, and 1 = Extremely disliked. Overall taste, Banana taste, Milk taste, Sweetness, Acidity, and Bitterness on a continuous 11-point intensity scale, where 0 = Imperceptible and 10 = Extremely pronounced. • Statistical analysis: ANOVA (by block) with Duncan's post-hoc test Petition 870260041436, dated 04 / 05 / 2026, page 27 / 51 19 / 34 • Sample size ~ 59.15 mL (2.0 oz) in a clear, lidded plastic cup of 103.51 mL (3.5 oz) • Serving temperature Refrigerated (~ 45 °F or 7.22 °C) • Instructions on consumption / by participants: Samples were served simultaneously. Participants were instructed to read the ingredient list and rate each sample. Table 11: Samples Control: Sugar and Stevia Extract (Reb A) Test: Sugar and Stevia Extract (Reb A) & GSG Ingredients Control Test-1 Low GSG levels Test-2 High GSG levels Milk (with 2% fat) 95.6% 95.6% 95.6% Sugar 4.2 4.2 4.2 Banana Flavor 0.20 0.20 0.20 Reb A 0.006 0.006 0.006 GSG 0.0175 0.035 TOTAL 100 100 100

[0048] In the present study, thirty-two members of the consumer panel evaluated three samples of banana-flavored dairy beverage for overall acceptance and the intensities of the attributes (overall flavor, banana flavor, milk flavor, sweetness, intensity of aftertaste). The three samples were: a control sample sweetened with sugar and stevia extract (Reb A) and not containing glycosyl steviol glycosides (GSG), and two test samples sweetened with sugar and stevia extract (Reb A) containing glycosyl steviol glycosides (GSG). One of the test samples contained a low amount of GSG (0.0175%) and the other contained a high amount of GSG (0.035%). The objective of the test was to determine if the addition of glycosyl steviol glycosides affects / improves the flavor profile of a banana-flavored dairy beverage. The results indicated that: The test sample containing a high level of GSG had significantly Petition 870260041436, dated 04 / 05 / 2026, page 28 / 51 20 / 34 the much more intense banana flavor, sweetness intensity, and delay in the appearance of the dairy flavor note than the control sample (with 95% confidence). - The test sample containing a low level of GSG also contributed to the greater intensity of the Banana flavor (with > 90% confidence). - There was no significant difference in overall acceptance, intensity of the dairy flavor, appearance of the banana flavor, and aftertaste between the test and control samples. Example 6: Bakery / baked goods with Glycosyl Steviol Glycosides

[0049] To evaluate flavor modification in baked goods, a range of GSG concentrations (0 to 5,000 ppm) were used with a Lemon and Poppy Seed muffin formula. The aim was to determine whether the addition of glycosyl steviol glycosides has an effect on key flavor attributes and / or improves flavor perception in various food applications, especially in various baked goods. Specifically, the aim was to determine whether the flavor profile and overall acceptance of a control sample of Lemon and Poppy Seed muffin (which does not contain glycosyl steviol glycosides) differs from a test sample of the same muffin (containing glycosyl steviol glycosides). Although the test was performed with muffins, the results are also relevant to all baked goods, not limited to cookies, cakes, pastries, bread, etc.Following preliminary sensory tests, it was found that GSG modified the flavor and sweetness profile at all concentrations. The preferred GSG concentration is in the range of approximately 0 to 5,000 ppm, more preferably in the range of approximately 100 to 3,000 ppm, and, primarily, in the range of approximately 100 to 2,000 ppm.

[0050] For detailed sensory testing, two were selected Petition 870260041436, dated 04 / 05 / 2026, page 29 / 51 21 / 34 samples to test against the control sample. The methodology for the detailed sensory tests is indicated below in Table 12. The formulas for the control and test samples are indicated below in Table 13. Table 12: Methodology • Number of sessions: 1 • Number of participants: 35 • Test design: Balanced, randomized within a pair. Blinded. • Sensory Test Method: Intensity and Acceptance Ratings • Environmental Condition: Standard booth lighting • Attributes and Scales: Overall acceptance on a 9-point hedonic scale, where 9 = Extremely liked, 5 = Neither liked nor disliked, and 1 = Extremely disliked; Overall taste, Lemon flavor, Sweetness, Acidity, and Bitterness on an 11-point continuous intensity scale, where 0 = Imperceptible and 10 = Extremely pronounced • General Open-Ended Comments • Gender and Age • Statistical Analysis: ANOVA (by block) with Duncan's post-hoc test • Sample Size: One 1.4-gram muffin in a 103.51 mL (3.5 oz) plastic soufflé cup • Consumption Temperature: Room temperature (~68°F or 20°C) • Instructions on Consumption / by Participants: Samples were served simultaneously.Participants were instructed to read the list of ingredients and evaluate each sample. Table 13: The samples Ingredients Control (%) Test (%) AP Flour 24.37 24.35 Milk 24.01 23.99 Sugar 23.16 23.14 Vegetable Oil 14.79 14.78 Petition 870260041436, dated 04 / 05 / 2026, page 30 / 51 22 / 34 Eggs 9.54 9.53 Poppy Seeds 1.00 1.00 Lemon Flavor 0.83 0.83 Salt 0.72 0.71 Lemon Juice 0.59 0.59 Vanilla Extract 0.52 0.52 Yeast 0.48 0.48 GSG 0.07 TOTAL 100.00 100.00

[0051] In the present study, thirty-five members of the consumer panel evaluated two samples of lemon and poppy seed muffins for overall acceptance and the intensities of the attributes (overall flavor, lemon flavor, sweetness, intensity of acidity and bitterness). The two samples were: 1) a control sample that does not contain glycosyl steviol glycosides (GSG); and 2) a sample containing GSG. The objective of the test was to determine if the addition of glycosyl steviol glycosides affects / improves the flavor profile of a lemon and poppy seed muffin. The results indicated that: The test sample (containing glycosyl steviol glycosides) had significantly more intense overall flavor and sweetness than the control (with 90% confidence). - There was no significant difference in overall flavor acceptance, lemon flavor intensity, acidity intensity, or bitterness intensity between the two samples (90% confidence). Directionally, the test sample was more generally accepted and had a more intense lemon flavor than the control (p-value = 0.124 and 0.190, respectively). Based on participant feedback, the control sample had less lemon flavor than the test sample. Example 7: Jams and Fruit / Vegetable Preparations with Glycosyl Steviol Glycosides Petition 870260041436, dated 04 / 05 / 2026, page 31 / 51 23 / 34

[0052] To evaluate the flavor modification of fruit / vegetable jams and fruit preparations, a range of GSG concentrations (0 to 5,000 ppm) was used with a Strawberry spreadable topping formula. The aim was to determine whether the addition of glycosyl steviol glycosides has an effect on key flavor attributes and / or enhances flavor perception in various food applications, especially in fruit or vegetable preparations. Specifically, the aim was to determine whether the flavor profile and overall acceptance of a control sample of Strawberry topping (containing no GSG) differ from a test sample of the same topping (containing GSG). Although the test was performed with Strawberry jam / preparations / topping, the results are also relevant to fruit and vegetable preparations, not limited to fruits (Banana, all berries, Mango, etc.) and vegetables (celery, artichoke, zucchini, avocado, etc.).After preliminary sensory tests, it was found that the flavor and sweetness profile were consistent at all concentrations. The GSG concentration is preferably in the range of approximately 0 to 5,000 ppm, more preferably in the range of approximately 1,000 to 4,000 ppm, and, primarily, in the range of approximately 2,000 to 3,000 ppm. For detailed sensory tests, two samples were selected to test against the control sample. The methodology for the detailed sensory tests is indicated below in Table 14. The formulas for the control and test samples are indicated below in Table 15. Table 14: Methodology • Number of sessions: 1 • Number of participants: 28 • Test design: Balanced, randomized within the pair. Blinded. • Sensory test method: Intensity and acceptance ratings • Environmental condition: Standard booth lighting • Attributes and scales Petition 870260041436, dated 04 / 05 / 2026, page 32 / 51 24 / 34 Overall acceptance on a 9-point hedonic scale, where 9 = Extremely liked, 5 = Neither liked nor disliked, and 1 = Extremely disliked. Overall taste, Fresh Strawberry Flavor, Sweetness, Acidity, and Bitterness on an 11-point continuous intensity scale, where 0 = Imperceptible and 10 = Extremely pronounced. • General Open Comments • Gender and age • Statistical Analysis ANOVA (by block) with Duncan's post-hoc test • Sample size ~ 14.79 mL (0.5 oz) in a 29.57 mL (1 oz) clear plastic cup with lid • Consumption temperature Refrigerated (~ 45 °F or 7.22 °C) • Instructions on consumption / by Participants: Samples were served simultaneously. Participants were instructed to read the ingredient list and rate each sample. Table 15: The samples Ingredients Control (%) Test (%) Sugar 39.23 39.18 Water 29.43 29.39 Strawberry Pieces 15.44 15.42 Strawberry Juice 15.44 15.42 Pectin A 0.26 0.26 Potassium Benzoate 0.10 0.10 RebA 0.06 0.06 Citric Acid 0.05 0.05 GSG 0.022 TOTAL 100.00 100.00

[0053] In this study, twenty-eight members of the consumer panel evaluated two samples of strawberry-flavored topping with reduced sugar for overall acceptance and intensity of attributes (overall flavor, strawberry flavor, sweetness, intensity of acidity and bitterness). The two samples were: 1) a control sample sweetened with sugar and Rebaudioside A, containing no glycosides Petition 870260041436, dated 04 / 05 / 2026, page 33 / 51 1) a test sample of 25 / 34 glycosyl steviol (GSG); and 2) a test sample sweetened with sugar and Rebaudioside A containing GSG. The aim of the test was to determine if the addition of GSG affects / improves the flavor profile of the strawberry-flavored coating. The results indicated that: - There was no significant difference in overall flavor acceptance, overall flavor intensity, Fresh Strawberry flavor intensity, sweetness intensity, or acidity intensity (with 90% confidence). The test sample was significantly less bitter than the control (with 90% confidence). Example 8: Yogurt with GSG

[0054] To evaluate the flavor modification in flavored and unflavored yogurts, a series of GSG concentrations (0 to 1,000 ppm) were used with a vanilla-flavored yogurt purchased from a local store. Although the test was conducted with vanilla-flavored yogurt, the results are also relevant to all other unflavored and flavored fermented dairy products, such as cheese, yogurt (with or without fat), liquid yogurts, smoothies, yogurt with fruit preparations, not limited to fruit (banana, all berries, mango, etc.). After preliminary sensory tests, it was found that GSG modified the flavor and sweetness profile at all concentrations. The GSG concentration is preferably in the range of about 0 to 1,000 ppm, more preferably in the range of about 50 to 500 ppm, and, primarily, in the range of about 100 to 400 ppm.

[0055] Three samples of vanilla-flavored yogurt (generic brand) were sweetened with sugar, sugar + Reb A, and sugar + Reb A + GSG. The amount of GSG was 220 ppm. The twelve-member panel found that GSG increased the sweetness profile and positively impacted the flavor profile. GSG helped harmonize the sweetness profile and directionally helped reduce the lingering taste of Reb A. Petition 870260041436, dated 04 / 05 / 2026, p. 34 / 51 26 / 34 Example 9: Lime-Lemon Carbonated Soft Drink with GSG

[0056] To evaluate flavor modification in a flavored carbonated soft drink (CSD), a range of GSG concentrations (0 to 1,000 ppm) were used in lime-lemon flavored CSDs. Although the test was performed on lime-lemon flavored CSDs with reduced sugar, the results are also relevant to all other flavored soft drinks (Cola, Orange, Grape, Passion Fruit, Berries group, Mango, etc.) with all sugar levels and diet (sugar-free) products. After preliminary sensory tests, it was found that GSG modified the flavor and sweetness profile at all concentrations. The GSG concentration is preferentially in the range of about 0 to 1,000 ppm, more preferably in the range of about 100 to 500 ppm, and, primarily, in the range of about 200 to 400 ppm.

[0057] Three samples of CSDs were produced with sugar, sugar + Reb A, and sugar + Reb A + GSG. The amount of GSG was 310 ppm. The sample with GSG directionally improved the sweetness profile, increased the Lemon note, and reduced the bitterness and aftertaste compared to the sample sweetened only with sugar and Reb A. Example 10. Chocolate drink with GSG

[0058] To evaluate the flavor modification of flavored dairy beverages, a range of GSG concentrations (0 to 1,000 ppm) were used with chocolate milk formulations sweetened with sugar and / or high fructose corn syrup (HFCS) and stevia. Although the test was conducted with a chocolate-flavored dairy beverage with reduced sugar, the results are also relevant to all other dairy beverages, with different fat levels or fat-free, flavored with different flavors (Strawberry, Blueberry, Mango, etc.), with different sugar levels including dietetic (sugar-free) products. After preliminary sensory tests, it was found that GSG modified the flavor and sweetness profile at all concentrations. The GSG concentration is Petition 870260041436, dated 04 / 05 / 2026, p. 35 / 51 27 / 34 preferably in the range of about 0 to 1,000 ppm, more preferably in the range of about 25 to 500 ppm, and, as a priority, in the range of about 50 to 400 ppm.

[0059] One of the test samples was sweetened with a mixture of HFCS 42, sugar and Reb A, the other test sample also included 337 ppm of GSG. GSG improved the Chocolate flavor, the milky note and the sweetness profile. Example 11: Coating a Baked Goods Product with GSG

[0060] To evaluate the flavor modification in toppings and jams used in bakery products, a concentration range of GSG (0 to 0.5%) was used in vanilla-flavored cake topping formulations sweetened with sugar and / or high-fructose corn syrup (HFCS) and stevia. Although the test was performed on a reduced-sugar topping, the results are also relevant for all other toppings, with different fat levels or no fat, and different flavors (Chocolate, Strawberry, Blueberry, Mango, etc.) with different sugar levels, including products with no added sugar. After preliminary sensory tests, it was found that GSG modified the sweetness profile and flavor at all concentrations. The GSG concentration is preferably in the range of about 0 to 0.5%, more preferably in the range of about 0.1 to 0.4%, and, primarily, in the range of about 0.2 to 0.3%.

[0061] As an example, a typical sensory test was carried out where GSG was added to a reduced-sugar cake frosting. The amount of GSG was 0.23%, which increased the sweetness profile to be most similar to sugar, as well as promoting the vanilla flavor. Example 12: Snacks with GSG

[0062] To evaluate flavor modification in snacks and cereal / oilseed products, a range of GSG concentrations (0 to 0.5%) were used in cinnamon-flavored coated almonds sweetened with sugar and / or high fructose corn syrup (HFCS). Although the test was carried out Petition 870260041436, dated 04 / 05 / 2026, p. 36 / 51 28 / 34 in oilseeds fully coated with sugar, the results are also relevant for all other coatings used for snacks, cereals, confections with different moisture and fat levels (or no fat) and different flavors (Chocolate, Cinnamon, Hazelnut, Maple, Brown Sugar, Strawberry, Blueberry, Mango, etc.) with different sugar levels, including products with no added sugar. After preliminary sensory tests, it was found that GSG modified the sweetness profile and flavor at all concentrations. The GSG concentration is preferably in the range of about 0 to 0.5%, more preferably in the range of about 0.05 to 0.4%, and, primarily, in the range of about 0.1 to 0.3%.

[0063] As an example, two coated almond snacks were prepared, where the test sample had reduced sugar. GSG was added in the amount of 0.19% to the test sample. GSG promoted harmonization of sweetness and increased cinnamon flavor in the test sample.

[0064] GSG can also be added to confectionery product formulations for the purpose of modifying the flavor of confectionery products, such as, but not limited to, hard-boiled candies, soft-textured candies, and chocolates. Example 13: Synergy of GSG Sweetness with Sugar

[0065] To evaluate the detection of GSG sweetness, a series of samples was produced with NSF-O2 (Natural Sweet Flavor #2) in acidified water (pH = 3.3, pH adjusted with citric acid). NSF-O2 contains GSG and approximately 15% to 20% dextrin, and is available from PureCircle, 915 Harger Road, Suite 250, Oak Brook, Illinois 60523, USA.

[0066] Aqueous solutions of NSF-O2 at various concentrations were prepared using mineral water that was acidified to a pH of approximately 3.5. The pH was adjusted using a 1% citric acid solution within a restricted range. The NSF-O2 concentrations ranged from 0 to 1,000 ppm. The samples were Petition 870260041436, dated 04 / 05 / 2026, p. 37 / 51 29 / 34 evaluated by the assessors at room temperature (21.1 - 22.2 °C (70 - 72 °F)).

[0067] The evaluators were 10 participants who were previously qualified for their acuity of taste and trained in the use of a sweetness intensity rating scale. The evaluations were performed in duplicate, using the same panel members (n = 20). Before conducting the study, the evaluators were presented with sugar controls prepared with acidified water for the intensity rating on the ballot, referencing 2, 4, 6, and 8 on the rating scale. These solutions were provided to the evaluators in order to refresh their memory regarding the intensity ratings.

[0068] Samples were given to evaluators sequentially and coded with three-digit numbers. The order of sample presentation was randomized to avoid bias. A five-minute rest period was provided between samples. Water and unsalted biscuits were provided to cleanse the palate. Evaluators could not detect any sweetness below 150 ppm of NSF-O2. The equivalent sweetness values ​​(SEV) of the different NSF-O2 solutions are shown in FIG. 6.

[0069] Similarly, a sensory test was conducted to quantify the synergy between NSF-O2 and sugar at 8% sucrose-equivalent sweetness in acidified water (pH approximately 3.5) at various levels of sugar reduction. The pH was adjusted using a 1% citric acid solution within a restricted range. The sugar was reduced by adding the required level of NSF-O2 until reaching 8% sugar-equivalent sweetness. As shown in FIG. 6, the sensory evaluation shows that the addition of NSF-O2 contributes to additional sweetness in the presence of sugar, even at a lower level of NSF-O2 (0 to 150 ppm NSF-O2 in acidified water, without sugar), it does not contribute any detectable level of Petition 870260041436, dated 04 / 05 / 2026, pp. 38 / 51 30 / 34 sweetness. The synergy between sugar and NSF-O2 is the difference in the SEV contribution of NSF-O2 with and without sugar. The concentration of NSF-O2 is preferably greater than about 25 ppm, more preferably greater than about 100 ppm, and ideally in the range of about 500 ppm to 1,000 ppm.

[0070] Therefore, the present invention shows that GSG not only modifies the flavor, but can also increase the sweetness in the presence of other sweeteners. This sweetness enhancement is caused by the synergy between GSG and other sweeteners. Example 14: Synergy between GSG and HFCS

[0071] The objective of this work was to develop the same sweetness and taste of high fructose corn syrup (HFCS) 55 in an aqueous solution with an equal amount of HFCS 42 plus GSG. HFCS 55 contains a total of 77% dry solids, and 55% of these dry solids is fructose. HFCS 42 contains 71% dry solids, and 42% of these dry solids is fructose. The equivalent sweetness values ​​(SEVS) of HFCS 55 and HFCS 42 are 0.99 and 0.91, respectively.

[0072] To match a similar sweetness and taste profile to HFCS 55, samples were tested including different amounts of GSG from 0 to 500 ppm and different bio-gums (xanthan gum, gum arabic, CMC, guar gum, locust bean gum, pectin), or polysaccharides (maltodextrin, oligosaccharides, resistant maltodextrin), or polyols. All combinations of GSG and bulking agent (to aid taste) provided the desired sweetness and taste that matched HFCS 55. The GSG concentration is preferably in the range of about 0 to 500 ppm, more preferably in the range of about 25 to 300 ppm, and, ideally, in the range of about 50 ppm to 200 ppm. However, the best solution was the combination of xanthan gum and GSG (shown in Table 16). A similar analysis was performed on a lime-lemon CSD application, which had a combination of GSG and xanthan gum to provide similar sweetness and taste. Petition 870260041436, dated 04 / 05 / 2026, pp. 39 / 51 31 / 34 to HFCS 42, as found in the formulation with HFC 55. Table 16 Ingredients: HFCS 55, HFCS 42, Water 87.02, HFCS 55 12.98, HFCS 42 12.98, Xanthan Gum 0.0085, GSG 0.0068, Brix 10 9.30 TOTAL 100 g 100 g - It was found that when xanthan gum was added together with GSG, the perception of taste was improved, as well as the overall flavor profile. The addition of GSG made the sample taste more like a sugar-based product. Sensory results:

[0073] Sensory analysis (discrimination test) was performed to determine the difference between beverage samples with HFCS 55 and HFCS 42. A triangle test was conducted with 19 participants. Only 4 identified the correct sample, two of whom were guessing. The two who correctly identified the odd sample indicated that the differences were due to HFCS 42 being less acidic and more flavorful; both also mentioned that HFCS 55 was slightly less sweet than HFCS 42.

[0074] Although the tests were carried out using high-fructose corn syrups, the results are not limited to high-fructose syrups made from corn. The results are also applicable to high-fructose syrups produced from other carbohydrate sources, such as, but not limited to, wheat, barley, cassava, rice, and potatoes. Example 15: Synergy of GSG with Other Non-Caloric Sweeteners

[0075] GSG was tested with several natural sweeteners (Reb A, SG95 Petition 870260041436, dated 04 / 05 / 2026, pp. 40 / 51 32 / 34 and PureCircle Alpha (derived from stevia extract) and synthetic sweeteners (sucralose, acesulfame-K, cyclamate, and aspartame) to investigate the synergy between GSG and high-intensity sweeteners. While GSG modifies the flavor profile, it also shows a different degree of synergy with high-intensity sweeteners. As an example, to estimate the synergy between GSG and stevia, NSF-02 (GSG + dextrin) was mixed with a required amount of PureCircle alpha or Reb A 97 in acidic solution (pH = 3.8) to achieve 8% sugar-equivalent sweetness as shown in Table 17. Alpha is a mixture of selected steviol glycosides, as described in International Patent Application No. PCT / US 2012 / 024,722, filed February 10, 2012, entitled Stevia Composition, and is available from PureCircle, 915 Harger Road, Suite 250, Oak Brook, Illinois 60523, USA. Reb A 97 is also available from PureCircle.The synergy was calculated as the reduction of stevia sweeteners (Alpha or Reb A) by the addition of different levels of NSF-O2, as shown in FIG. 7. Table 17 Control ppm ppm ppm ppm ppm ppm NSF-02 0 25 50 100 150 200 250 Reb A 345 340 340 338 286 272 260 Alpha 400 350 345 320 280 252 235 Synergy with Reb A 0 5 5 7 59 73 85 Synergy with Alpha 0 50 55 80 120 148 165

[0076] It was found that Alfa shows improved sweetness in the presence of a very small amount of NSF-O2 (25 ppm or less), whereas above 100 ppm, NSF-O2 had to be added to achieve any synergy with Reb A, as shown in FIG. 7. Note that the detection level of NSF-O2 is about 150 ppm, as shown in FIG. 6. In an Alfa solution, the NSF-O2 concentration is preferably greater than about 10 ppm, more preferably greater than about 25 ppm, and, preferentially, greater than about Petition 870260041436, dated 04 / 05 / 2026, pp. 41 / 51 33 / 34 of 100 ppm. In a solution with Reb A, the concentration of NSF-O2 is preferably greater than about 100 ppm, more preferably greater than about 150 ppm, and, most importantly, greater than about 200 ppm. Example 16: GSG as a Taste modifier

[0077] GSG works with sugar, HFCS, and other natural sweeteners to provide a better taste and sweetness profile in beverages. Erythritol is used in stevia-based drinks to provide some sweetness, but mainly to contribute to the taste that is lacking when a high amount of sugar is replaced by the high-intensity sweetener. A study was conducted to investigate the amount of erythritol that can be replaced by GSG in non-carbonated beverages.

[0078] Sample preparation: A number of acidified beverage samples were subjected to a sweetness level of 8 Brix with 200 ppm of Reb A and the combination of erythritol and GSG as shown in Table 18. Table 18 Erythritol Concentration (%) 3.5 2.5 1.5 3 2.75 E E6 E7 E8 E9 Reb A 0.1 0.1 0.1 0.1 0.1 GSG 0.044 0.0875 0.022 0.033 Erythritol 17.5 12.5 7.5 15 13.75 Citric Acid 0.25 0.25 0.25 0.25 0.25 Water 482.2 487.1 492.1 484.6 485.9 Total 500 500 500 500 500 Sensory Evaluation: 1. In comparing samples E, E6, and E7, it was found that E6 is very close to the control (E) in terms of flavor and overall taste, and slightly less sweet than the control. E7, which had a higher water content (very watery). 2. A triangular test with samples E and E6 was conducted over two days, with a total of 12 panel members. Five of them detected the sample. Petition 870260041436, dated 04 / 05 / 2026, pp. 42 / 51 34 / 34 correct. The conclusion was that the panel members were able to detect the difference, therefore the test failed. 3. A preliminary test with E, E8, E9 was then conducted, followed by a triangle test with E and E9. Three of the ten panel members were able to identify the correct sample.

[0079] Conclusion: GSG can reduce the use of erythritol by 20-30% (from 3.5% to 2.75-3%) in beverages without any sacrifice to taste or flavor. In the flavor system studied, the GSG concentration is preferably greater than about 10 ppm, more preferably greater than about 20 ppm, and ideally in the range of about 30 ppm to about 200 ppm. In some flavor systems, GSG can replace more erythritol in order to provide a balanced, harmonized sweet taste.

[0080] Although the present invention and its advantages have been described in detail, it should be understood that various changes, substitutions, and alterations may be made hereto without departing from the spirit and scope of the invention, as defined by the appended claims. Furthermore, the scope of this patent application shall not be limited to the particular embodiments of the invention described in the specification. A person skilled in the art will readily understand from the disclosure of the present invention the compositions, devices, processes, methods, and steps, now existing or to be developed hereafter, which substantially perform the same function or achieve substantially the same result as the corresponding embodiments described herein, which may be used in accordance with the present invention. Petition 870260041436, dated 04 / 05 / 2026, pp. 43 / 51

Claims

1 / 1 CLAIMS 1. Flavor and taste enhancing composition CHARACTERIZED in that it comprises 95 to 19% mono- and diglycosylated steviol glycosides, 0 to 60% glycosylated steviol glycosides G3 to G9, 0 to 16% glycosylated steviol glycosides G10 to G20.

2. Flavor and taste enhancing composition, according to claim 1, CHARACTERIZED in that at least one glucose unit occurs at the C-19 position of the steviol glycoside.

3. Food or beverage product with an intense taste and flavor profile CHARACTERIZED in that it comprises the flavor and taste-enhancing composition as defined in claim 1 or 2.

4. Method for increasing the taste and flavor intensity of a food or beverage product CHARACTERIZED in that it comprises the step of adding the flavor and taste-enhancing composition, as defined in claim 1 or 2, to the food or beverage product. Petition 870260041436, dated 04 / 05 / 2026, pp. 44 / 51

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