Patent Publication Number: US-2021186070-A1

Title: Stevia extract containing selected steviol glycosides as flavor, salty and sweetness profile modifier

Description:
RELATED APPLICATIONS 
     This application is a divisional application of U.S. patent application Ser. No. 15/619,107, filed on Jun. 9, 2017, which is a continuation-in-part application of U.S. patent application Ser. No. 14/896,022, filed on Dec. 4, 2015, which is a national phase application of International Application No. PCT/US2014/041548, filed on Jun. 9, 2014, and claims the benefit of priority to U.S. Patent Application No. 61/832,451, filed on Jun. 7, 2013, and U.S. Patent Application No. 61/942,331, filed on Feb. 20, 2014, the contents of which applications are incorporated by reference herein in their entirety. 
    
    
     FIELD OF THE INVENTION 
     The invention relates to the use of  Stevia  extracts as flavor modifiers that contain mixtures of steviol glycosides extracted from  Stevia rebaudiana  plant. This invention also relates to the application of the above-said  Stevia  extracts as sweetness profile modifier, not a sweetener, with other natural and artificial sweeteners. This invention also relates to the production and use of the above-mentioned  Stevia  extracts that can be used as flavor and sweetness profile modifier when used in food, beverage, and pharmaceutical products. 
     BACKGROUND 
     High intensity sweeteners possess sweetness level many times exceeding that of sucrose. They are essentially non-caloric and used widely in manufacturing of diet and reduced calorie food. Although natural caloric sweetener such as sucrose, fructose, and glucose provide the most desirable taste to consumers, they are caloric. High intensity sweeteners do not affect the blood glucose level and provide little or no nutritive value. 
     However, high intensity sweeteners that generally are used as substitutes for sucrose possess taste characteristics different than that of sugar, such as sweet taste with different temporal profile, maximal response, flavor profile, mouthfeel, and/or adaptation behavior than that of sugar. For example, the sweet taste of some high-potency sweeteners is slower in onset and longer in duration than that of sugar and thus changes the taste balance of a food composition. Because of these differences, usage of high-potency sweetener in replacing such a bulk sweetener as sugar in a food or beverage causes imbalance in temporal and/or flavor profile. If the taste profile of high-potency sweeteners could be modified to impart desired taste characteristics, it can provide low calorie beverages and food products with taste characteristics more desirable for consumers. To attain the sugar-like temporal and/or flavor profile, several ingredients have been suggested in different publications. 
     Non-limiting examples of synthetic sweeteners include sucralose, potassium acesulfame, aspartame, alitame, saccharin, neohesperidin dihydrochalcone synthetic derivatives, cyclamate, neotame, dulcin, suosan, N—[N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-L-α-aspartyl]-L-phenylalanine 1-methyl ester, N—[N-[3-(3-hydroxy-4-methoxyphenyl)-3-methylbutyl]-L-α-aspartyl]-phenylalanine 1-methyl ester, N—[N-[3-(3-methoxy-4-hydroxyphenyl)propyl]-L-α-aspartyl]-phenylalanine 1-methyl ester, salts thereof, and the like. 
     Non-limiting examples of natural high intensity sweeteners include Stevioside, Rebaudioside A, Rebaudioside B, Rebaudioside C, Rebaudioside E, Rebaudioside F, Steviolbioside, Dulcoside A, Rubusoside, mogrosides, brazzein, neohesperidin dihydrochalcone (NHDC), glycyrrhizic acid and its salts, thaumatin, perillartine, pernandulcin, mukuroziosides, baiyunoside, phlomisoside-I, dimethyl-hexahydrofluorene-dicarboxylic acid, abrusosides, periandrin, carnosiflosides, cyclocarioside, pterocaryosides, polypodoside A, brazilin, hernandulcin, phillodulcin, glycyphyllin, phlorizin, trilobatin, dihydroflavonol, dihydroquercetin-3-acetate, neoastilibin, trans-cinnamaldehyde, monatin and its salts, selligueain A, hematoxylin, monellin, osladin, pterocaryoside A, pterocaryoside B, mabinlin, pentadin, miraculin, curculin, neoculin, chlorogenic acid, cynarin, siamenoside and others. 
     High intensity sweeteners can be derived from the modification of natural high intensity sweeteners, for example, by fermentation, enzymatic treatment, or derivatization. 
     A growing number of consumers perceive the ability to control their health by enhancing their current health and/or hedging against future diseases. This creates a demand for food products with enhanced characteristics and associated health benefits, specifically a food and consumer market trend towards “whole health solutions” lifestyle. The term “natural” is highly emotive in the world of sweeteners and has been identified as one of key trust, along with “whole grains”, “heart-healthy” and “low-sodium”. ‘Natural’ term is closely related to ‘healthier’. 
       Stevia rebaudiana  is a perennial shrub of the Asteraceae (Compositae) family native to certain regions of South America. The leaves of the plant contain from 10 to 20% of diterpene glycosides, which are around 150 to 450 times sweeter than sugar. The leaves have been traditionally used for hundreds of years in Paraguay and Brazil to sweeten local beverages, foods and medicines. 
     At present there are more than 230  Stevia  species with significant sweetening properties. The plant has been successfully grown under a wide range of conditions from its native subtropics to the cold northern latitudes. 
     Steviol glycosides have zero calories and can be used wherever sugar is used. They are ideal for diabetic and low calorie diets. In addition, the sweet steviol glycosides possess functional and sensory properties superior to those of many high potency sweeteners. 
     The extract of  Stevia rebaudiana  plant 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 compose approximately 10%-20% of the total dry weight. Typically, on a dry weight basis, the four major glycosides found in the leaves of  Stevia  are Dulcoside A (0.3%), Rebaudioside C (0.6%), Rebaudioside A (3.8%) and Stevioside (9.1%). Other glycosides identified in  Stevia  extract include Rebaudioside B, C, D, E, and F, Steviolbioside and Rubusoside ( FIG. 1 ). 
     The chemical structures of the diterpene glycosides of  Stevia rebaudiana  are presented in  FIG. 1 . The physical and sensory properties are well studied only for Stevioside and Rebaudioside A. The sweetness potency of Stevioside is around 210 times higher than sucrose, Rebaudioside A around 300 times, and Rebaudioside C and Dulcoside A around 30 times. The  Stevia  extract containing Rebaudioside A and Stevioside as major components showed sweetness potency around 250 times. Rebaudioside A and Rebaudioside D are considered to have most favorable sensory attributes of all major Steviol Glycosides (TABLE 1). 
     
       
         
           
               
               
               
               
               
               
               
             
               
                 TABLE 1 
               
               
                   
               
               
                   
                   
                   
                 Mol. 
                 Solubility in 
                 Relative 
                   
               
               
                 Name 
                 Formula 
                 T Melt , ° C. 
                 Weight 
                 water, % 
                 sweetness 
                 Quality of taste 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 Steviol 
                 C 20 H 30 O 3   
                 212-213 
                 318.45 
                 ND 
                 ND 
                 Very bitter 
               
               
                 Steviolmonoside 
                 C 26 H 40 O 8   
                 ND 
                 480.58 
                 ND 
                 ND 
                 ND 
               
               
                 Stevioside 
                 C 38 H 60 O 18   
                 196-198 
                 804.88 
                 0.13 
                 210 
                 Bitter 
               
               
                 Rebaudioside A 
                 C 44 H 70 O 23   
                 242-244 
                 967.01 
                 0.80 
                 200-400 
                 Less Bitter 
               
               
                 Rebaudioside B 
                 C 38 H 60 O 18   
                 193-195 
                 804.88 
                 0.10 
                 150 
                 Bitter 
               
               
                 Rebaudioside C 
                 C 44 H 70 O 22   
                 215-217 
                 951.01 
                 0.21 
                 30 
                 Bitter 
               
               
                 Rebaudioside D 
                 C 50 H 80 O 28   
                 248-249 
                 1129.15 
                 1.00 
                 220 
                 Like sucrose 
               
               
                 Rebaudioside E 
                 C 44 H 70 O 23   
                 205-207 
                 967.01 
                 1.70 
                 170 
                 Like sucrose 
               
               
                 Rebaudioside F 
                 C 43 H 68 O 22   
                 ND 
                 936.99 
                 ND 
                 ND 
                 ND 
               
               
                 Dulcoside A 
                 C 38 H 60 O 17   
                 193-195 
                 788.87 
                 0.58 
                 30 
                 Very bitter 
               
               
                 Steviolbioside 
                 C 32 H 50 O 13   
                 188-192 
                 642.73 
                 0.03 
                 90 
                 Unpleasant 
               
               
                 Rubusoside 
                 C 32 H 50 O 13   
                 ND 
                 642.73 
                 ND 
                 110 
                 Very bitter 
               
               
                   
               
            
           
         
       
     
     In addition to the commercially known steviol glycosides (Table 1), several new steviol glycosides (glycosylated diterpene) have been found in  Stevia  leaf extracts, as shown in Table 2a. 
     
       
         
           
               
             
               
                 TABLE 2a 
               
             
            
               
                   
               
               
                 Summary of formula and R-groups of identified steviol glycosides 
               
               
                 (see FIG. 1 for backbone structure) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                 Trivial 
                 Mol. 
                   
                   
                   
               
               
                 # 
                 Common name 
                 formula 
                 Wt. 
                 R 1   
                 R 2   
                 Reference 
               
               
                   
               
            
           
           
               
            
               
                 1. Steviol + Glucose (SvGn) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1.1 
                 Steviolmonoside 
                 SvG1 
                   481 
                 H 
                 Glcβ1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                   
                   
                 (2010) 
               
               
                 1.2 
                 Steviolmonoside 
                 SvG1 
                   481 
                 Glcβ1- 
                 H 
                 Gardena et 
               
               
                   
                 A 
                   
                   
                   
                   
                 al. (2010) 
               
               
                 1.3 
                 Rubusoside 
                 SvG2 
                   643 
                 Glcβ1- 
                 Glcβ1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                   
                   
                 (2010) 
               
               
                 1.4 
                 Steviolbioside 
                 SvG2 
                   643 
                 H 
                 Glcβ(1-2)Glcβ1- 
                 Kohda et 
               
               
                   
                   
                   
                   
                   
                   
                 al. (1976) 
               
               
                 1.5 
                 Stevioside 
                 SvG3 
                   805 
                 Glcβ1- 
                 Glcβ(1-2)Glcβ1- 
                 Bridel &amp; 
               
               
                   
                   
                   
                   
                   
                   
                 Lavielle 
               
               
                   
                   
                   
                   
                   
                   
                 (1931) 
               
               
                 1.6 
                 Stevioside A 
                 SvG3 
                   805 
                 Glcβ(1- 
                 Glcβ1- 
                 Wu et al. 
               
               
                   
                   
                   
                   
                 2)Glcβ1- 
                   
                 (2012) 
               
               
                 1.7 
                 Rebaudioside B 
                 SvG3 
                   805 
                 H 
                 Glcβ(1- 
                 Kohda et 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 al. (1976) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 1.8 
                 Rebaudioside G 
                 SvG3 
                   805 
                 Glcβ1- 
                 Glcβ(1-3)Glcβ1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                   
                   
                 (2010) 
               
               
                 1.9 
                 Stevioside B 
                 SvG3 
                   805 
                 Glcβ(1- 
                 Glcβ1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                 3)Glcβ1- 
                   
                 &amp; Zamora 
               
               
                   
                   
                   
                   
                   
                   
                 (2014) 
               
               
                 1.10 
                 Rebaudioside E 
                 SvG4 
                   967 
                 Glcβ(1- 
                 Glcβ(1-2)Glcβ1- 
                 Sakamoto 
               
               
                   
                   
                   
                   
                 2)Glcβ1- 
                   
                 et al. 
               
               
                   
                   
                   
                   
                   
                   
                 (1977a) 
               
               
                 1.11 
                 Rebaudioside A 
                 SvG4 
                   967 
                 Glcβ1- 
                 Glcβ(1- 
                 Kohda et 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 al. (1976) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 1.12 
                 Rebaudioside A2 
                 SvG4 
                   967 
                 Glcβ1- 
                 Glcβ(1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                   
                 6)Glcβ(1- 
                 &amp; Prakash 
               
               
                   
                   
                   
                   
                   
                 2)Glcβ1- 
                 (2011d) 
               
               
                 1.13 
                 Rebaudioside D 
                 SvG5 
                 1 129 
                 Glcβ(1- 
                 Glcβ(1- 
                 Sakamoto 
               
               
                   
                   
                   
                   
                 2)Glcβ1- 
                 2)[Glcβ(1- 
                 et al. 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                 (1977a) 
               
               
                 1.14 
                 Rebaudioside I 
                 SvG5 
                 1 129 
                 Glcβ(1- 
                 Glcβ(1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                 3)Glcβ1- 
                 2)[Glcβ(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 1.15 
                 Rebaudioside L 
                 SvG5 
                 1 129 
                 Glcβ1- 
                 Glcβ(1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                   
                 6)Glcβ(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                   
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 1.16 
                 Rebaudioside Q2 
                 SvG5 
                 1 129 
                 Glcα(1- 
                 Glcβ(1-2)Glcβ1-  
                 Chaturvedula 
               
               
                   
                   
                   
                   
                 2)Glcα(1- 
                   
                 &amp; Prakash 
               
               
                   
                   
                   
                   
                 4)Glcβ1- 
                   
                 (2011c) 
               
               
                 1.17 
                 Rebaudioside Q 
                 SvG5 
                 1 129 
                 Glcβ1- 
                 Glcα(1- 
                 — 
               
               
                   
                   
                   
                   
                   
                 4)Glcβ(1- 
                   
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                   
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 1.18 
                 Rebaudioside I2 
                 SvG5 
                 1 129 
                 Glcβ1- 
                 Glcα(1-  
                 Chaturvedula 
               
               
                   
                   
                   
                   
                   
                 3)Glcβ(1- 
                 et al. 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 (2011c) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 1.19 
                 Rebaudioside Q3 
                 SvG5 
                 1 129 
                 Glcβ1- 
                 Glcα(1-  
                 Chaturvedula 
               
               
                   
                   
                   
                   
                   
                 4)Glcβ(1- 
                 et al. 
               
               
                   
                   
                   
                   
                   
                 3)[Glcβ(1- 
                 (2011c) 
               
               
                   
                   
                   
                   
                   
                 2)]Glcβ1- 
                   
               
               
                 1.20 
                 Rebaudioside I3 
                 SvG5 
                 1 129 
                 Glcβ(1- 
                 Glcβ(1-2)Glcβ1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                 2)[Glcβ(1- 
                   
                 et al. 
               
               
                   
                   
                   
                   
                 6)]Glcβ1- 
                   
                 (2011c) 
               
               
                 1.21 
                 Rebaudioside M 
                 SvG6 
                 1 291 
                 Glcβ(1- 
                 Glcβ(1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                 2)[Glcβ (1- 
                 2)[Glcβ(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                 3)]Glcβ1- 
                 3)]Glcβ1- 
                   
               
            
           
           
               
            
               
                 2. Steviol + Rhamnose + Glucose (SvR1Gn) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 2.1 
                 Dulcoside A 
                 SvR1G2 
                   789 
                 Glcβ1- 
                 Rhaα(1-2)Glcβ1- 
                 Kobayashi 
               
               
                   
                   
                   
                   
                   
                   
                 et al. 
               
               
                   
                   
                   
                   
                   
                   
                 (1977) 
               
               
                 2.2 
                 Dulcoside B 
                 SvR1G2 
                   789 
                 H 
                 Rhaα(1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 2.3 
                 Rebaudioside C 
                 SvR1G3 
                   951 
                 Glcβ1- 
                 Rhaα(1- 
                 Sakamoto 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 et al. 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                 (1977b) 
               
               
                 2.4 
                 Rebaudioside 
                 SvR1G3 
                   951 
                 Rhaα(1- 
                 Glcβ(1-2)Glcβ1- 
                 Purkayastha 
               
               
                   
                 C2 a   
                   
                   
                 2)Glcβ1- 
                   
                 (2016) 
               
               
                 2.5 
                 Rebaudioside S 
                 SvR1G3 
                   951 
                 Rhaα(1- 
                 Glcα (1- 
                 Ibrahim et 
               
               
                   
                   
                   
                   
                 2)Glcβ1- 
                 2)Glcβ1- 
                 al (2016) 
               
               
                 2.6 
                 Rebaudioside H 
                 SvR1G4 
                 1 112 
                 Glcβ1- 
                 Glcβ(1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                   
                 3)Rhaα(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                   
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 2.7 
                 Rebaudioside K 
                 SvR1G4 
                 1 112 
                 Glcβ(1- 
                 Rhaα(1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                 2)Glcβ1- 
                 2)[Glcβ(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 2.8 
                 Rebaudioside J 
                 SvR1G4 
                 1 112 
                 Rhaα(1- 
                 Glcβ(1-  
                 Ohta et al. 
               
               
                   
                   
                   
                   
                 2)Glcβ1- 
                 2)[Glcβ(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 2.9 
                 Rebaudioside N 
                 SvR1G5 
                 1 274 
                 Rhaα(1- 
                 Glcβ(1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 2)[Glcβ(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                 3)]Glcβ1- 
                 3)]Glcβ1- 
                   
               
               
                 2.10 
                 Rebaudioside O 
                 SvR1G6 
                 1 436 
                 Glcβ(1- 
                 Glcβ(1- 
                 Ohta et al. 
               
               
                   
                   
                   
                   
                 3)Rhaα(1- 
                 2)[Glcβ(1- 
                 (2010) 
               
               
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 3)]Glcβ1- 
                   
               
               
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
                   
               
               
                 2.11 
                 Rebaudioside 
                 SvR1G6 
                 1 436 
                 Glcβ(1- 
                 Glcβ(1- 
                 Purkayastha 
               
               
                   
                 O2 a   
                   
                   
                 4*)Rhaα(1- 
                 2)[Glcβ(1- 
                 (2016) 
               
               
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 3)]Glcβ1- 
                   
               
               
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
                   
               
               
                 2.12 
                 Rebaudioside 
                 SvR1G4 
                 1 112 
                 Glcβ(1- 
                 Rhaα(1- 
                 Purkayastha 
               
               
                   
                 K2 a   
                   
                   
                 6)Glcβ1- 
                 2)[Glcβ(1- 
                 (2016) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
            
           
           
               
            
               
                 3. Steviol + Xylose + Glucose (SvX1Gn) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 3.1 
                 Stevioside F 
                 SvX1G2 
                   775 
                 Glcβ1- 
                 Xylβ(1-2)Glcβ1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                   
                   
                 &amp; Prakash 
               
               
                   
                   
                   
                   
                   
                   
                 (2011b) 
               
               
                 3.2 
                 Rebaudioside F 
                 SvX1G3 
                   937 
                 Glcβ1- 
                 Xylβ(1- 
                 Starratt et 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 al. (2002) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 3.3 
                 Rebaudioside F2 
                 SvX1G3 
                   937 
                 Glcβ1- 
                 Glcβ(1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                   
                 2)[Xylβ(1- 
                 &amp; Prakash 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                 (2011b) 
               
               
                 3.4 
                 Rebaudioside F3 
                 SvX1G3 
                   937 
                 Xylβ(1- 
                 Glcβ(1-2)Glcβ1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                 6)Glcβ1- 
                   
                 et al. 
               
               
                   
                   
                   
                   
                   
                   
                 (2011d) 
               
               
                 3.5 
                 Rebaudioside R 
                 SvX1G3 
                   937 
                 Glcβ1- 
                 Glcβ(1- 
                 Ibrahim et 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1-3)] 
                 al (2016) 
               
               
                   
                   
                   
                   
                   
                 Xylβ1- 
                   
               
               
                 3.6 
                 Rebaudioside 
                 SvX1G4 
                 1 099 
                 Xylβ(1- 
                 Glcβ(1- 
                 Purkayastha 
               
               
                   
                 U a   
                   
                   
                 2)Glcβ1- 
                 2)[Glcβ(1- 
                 (2016) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 3.7 
                 Rebaudioside 
                 SvX1G4 
                 1 099 
                 Xylβ(1- 
                 Glcβ(1-2)Glcβ1- 
                 Purkayastha 
               
               
                   
                 U2 a   
                   
                   
                 2*)[Glcβ(1- 
                   
                 (2016) 
               
               
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
                   
               
               
                 3.8 
                 Rebaudioside 
                 SvX1G5 
                 1 261 
                 Glcβ(1- 
                 Xylβ(1- 
                 Purkayastha 
               
               
                   
                 V a   
                   
                   
                 2)[Glcβ(1- 
                 2*)[Glcβ(1- 
                 (2016) 
               
               
                   
                   
                   
                   
                 3)]Glcβ1- 
                 3)]Glcβ1- 
                   
               
               
                 3.9 
                 Rebaudioside 
                 SvX1G5 
                 1 261 
                 Xylβ (1- 
                 Glcβ(1- 
                 Prakash &amp; 
               
               
                   
                 V2 a   
                   
                   
                 2)[Glcβ(1- 
                 2)[Glcβ(1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                 3)]Glcβ1- 
                 3)]Glcβ1- 
                 (2013) 
               
            
           
           
               
            
               
                 4. Steviol + Arabinose + Glucose (SvA1Gn) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 4.1 
                 Rebaudioside 
                 SvA1G4 
                 1 098 
                 Glcβ(1- 
                 Glcβ(1-2)Glcβ1- 
                 Purkayastha 
               
               
                   
                 W a   
                   
                   
                 2)[Araβ(1- 
                   
                 (2016) 
               
               
                   
                   
                   
                   
                 3*)]Glcβ1 
                   
                   
               
               
                 4.2 
                 Rebaudioside 
                 SvA1G4 
                 1 098 
                 Araβ(1- 
                 Glcβ(1- 
                 Purkayastha 
               
               
                   
                 W2 a   
                   
                   
                 2*)Glcβ1 
                 2)[Glcβ(1- 
                 (2016) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 4.3 
                 Rebaudioside 
                 SvA1G4 
                 1 098 
                 Araβ(1- 
                 Glcβ(1- 
                 Purkayastha 
               
               
                   
                 W3 a   
                   
                   
                 6)Glcβ1- 
                 2)[Glcβ(1- 
                 (2016) 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
               
                 4.4 
                 Rebaudioside 
                 SvA1G5 
                 1 260 
                 Glcβ(1- 
                 Glcβ(1- 
                 Purkayastha 
               
               
                   
                 Y a   
                   
                   
                 2)[Araβ(1- 
                 2)[Glcβ(1- 
                 (2016) 
               
               
                   
                   
                   
                   
                 3*)]Glcβ1 
                 3)]Glcβ1- 
                   
               
            
           
           
               
            
               
                 5. Steviol + Fructose + Glucose (SvF1Gn) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 5.1 
                 Rebaudioside A3 
                 SvF1G3 
                   967 
                 Glcβ1- 
                 Glcβ(1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                   
                 2)[Fruβ(1- 
                 et al. 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                 (2011b) 
               
            
           
           
               
            
               
                 6. Steviol + galactose + Glucose (SvGa1Gn) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 6.1 
                 Rebaudioside 
                 SvGa1 
                 1 128 
                 Galβ(1- 
                 Glcβ(1- 
                 Purkayastha 
               
               
                   
                 T a   
                 G4 
                   
                 2*)Glcβ1 
                 2)[Glcβ(1- 
                 (2016){circumflex over ( )} 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                   
               
            
           
           
               
            
               
                 7. Steviol + de-oxy glucose + Glucose (SvdG1Gn) 
               
            
           
           
               
               
               
               
               
               
               
            
               
                 7.1 
                 Stevioside D 
                 SvdG1G2 
                   789 
                 Glcβ1- 
                 6-deoxyGlcβ(1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                   
                 2)Glcβ1- 
                 &amp; Prakash 
               
               
                   
                   
                   
                   
                   
                   
                 (2011a) 
               
               
                 7.2 
                 Stevisoide E 
                 SvdG1G3 
                   951 
                 Glcβ1- 
                 6-deoxyGlcβ(1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                   
                 2)[Glcβ(1- 
                 &amp; Prakash 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                 (2011a) 
               
               
                 7.3 
                 Stevioside E2 
                 SvdG1G3 
                   951 
                 6- 
                 Glcβ(1- 
                 Chaturvedula 
               
               
                   
                   
                   
                   
                 deoxyGlcβ1- 
                 2)[Glcβ(1- 
                 et al. 
               
               
                   
                   
                   
                   
                   
                 3)]Glcβ1- 
                 (2011e) 
               
               
                   
               
            
           
         
       
     
     Besides diterpene glycosides, a number of flavonoids, labdane diterpene, triterpenes, sterols, and volatile oils have also been found in the extracts of  Stevia rebaudiana,  collectively referred to as plant molecules, as shown in Table 2b. 
     
       
         
           
               
               
             
               
                 TABLE 2b 
               
               
                   
               
               
                 Chemical Classes 
                 Chemical Components 
               
               
                   
               
             
            
               
                 Monoterpenoids 
                 Borneol 
               
               
                 Diterpenoids 
                 Austroinulin, 6-0-acetyl austroinulin, 6-acetyl austroinulin 
               
               
                   
                 7-0-acetyl austroinulin, Sterebin A, B, C, D, E, F, G, H, Jhanol 
               
               
                 Triterpenoids 
                 Amyrin beta acetate 
               
               
                 Sesquiterpenes 
                 α-bergamotene, Bisabolene, β-bourbonene, δ-cadinene, γ-cadinene  
               
               
                 Essential oils 
                 β-caryophyllene, Trans β-tarnesene, α-humulene, δ-cadiene 
               
               
                   
                 caryophyllene oxide, Nerolidol, Linalol, α-terpineol, Terpinen-4-ol 
               
               
                 Sterol derivatives 
                 Stigmasterol , β-sitosterol, Campesterol 
               
               
                 Flavonoids 
                 Glucosy1-4′-O-apigenin, Glucosyl-7-O-luteolin, Rhamnosyl-3-O- 
               
               
                   
                 kaempferol, Quercetin, Glucosyl-3-O-quercetin, Arabinosyl-3-O- 
               
               
                   
                 quercetin, 5,7,3′-methoxyflavone, 3,6,4′-methoxyflavone, 
               
               
                   
                 Centaureidin, avicularin 
               
               
                   
               
            
           
         
       
     
     All steviol glycosides provide sweetness and other taste attributes at a higher than certain threshold level of concentrations in water. Below the threshold level of concentration, the steviol glycoside components and their mixtures as found in a typical non-limiting  Stevia  extract as shown below has no recognizable sweetness taste. But such  Stevia  extract below the threshold level of significant sweetness recognition show remarkable characteristics of sweet and flavor profile modification in food and beverage applications. 
     This invention relates to use of the following  Stevia  extracts (Table 3) with the varying level of different steviol glycosides and other  Stevia  plant-derived glycosides, the combination of which contributes no significant sweetness but modifies flavor and sweetness profile at certain concentration in typical food and beverage applications. 
     
       
         
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Minor 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Steviol 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 Glycosides 
               
            
           
           
               
               
               
               
            
               
                   
                 Steviol Glycosides*, % 
                   
                 and related 
               
            
           
           
               
               
               
               
               
               
               
               
               
               
               
               
               
               
               
            
               
                 Stevia 
                   
                   
                   
                 Reb 
                   
                 Dulco- 
                 Rubu- 
                   
                 Steviolbio- 
                   
                   
                   
                 TSG* 
                 plant 
               
               
                 Extracts 
                 Reb A  
                 Stevioside 
                 Reb D 
                 F 
                 Reb C 
                 side A 
                 soside 
                 Reb B 
                 side 
                 Reb E 
                 Reb N 
                 Reb O 
                 (%) 
                 molecules 
               
               
                   
               
               
                 PCS-5001 
                 10-20 
                 4-12 
                 1-4 
                 1-5 
                 10-25 
                 1-5 
                 1-4 
                 0.5-5 
                 0.5-5 
                 1-4 
                 0.5-4  
                 0.5-4 
                 45-65 
                 35-50 
               
               
                 PCS-1015 
                 18-25 
                 5-10 
                 8-20 
                 0-1 
                  1-3 
                 0-1 
                 0-1 
                 0.5-5 
                   0-1 
                 2-6 
                   4-8 
                   3-8 
                 55-65 
                 35-45 
               
               
                 PSB-5005 
                 15-30 
                 3-12 
                 1-10 
                 0-5 
                  5-15 
                 0-5 
                 0-5 
                   0-8 
                   0-5 
                 0-5 
                   0-6 
                   0-5 
                 45-70 
                 30-55% 
               
               
                   
               
               
                 *TSG or Total Steviol Glycosides contain the Steviol Glycosides that are recognized by Codex Alimentarius (a commission of FAO and WHO) and major regulatory authorities 
               
            
           
         
       
     
     The present invention also relates to the  Stevia  extracts that contain major steviol glycosides (Table 3) and other minor steviol glycosides and glycosylated diterpene derivatives (water soluble molecules). The non-limiting examples of such minor molecules are Reb E, Reb G, Reb H, Reb I, Reb K, Reb L, Reb M, Reb N, Reb O (M. Ohta, S. Sasa, A. Inoue, et al. “Characterization of Novel Steviol Glycosides from Leaves of  Stevia rebaudiana  Morita.”  J. Appl. Glycosci.,  57, 1 99-209 (201 0)). 
     The present invention is also directed to a method of making a specific  Stevia  extract composition, including: extracting steviol glycosides and other water soluble molecules from leaves of a  Stevia rebaudiana  plant, and separating the excess steviol glycosides than the amount and type of steviol glycosides required to contribute the taste and flavor modifying characteristics of the  Stevia  extract. 
     This invention combine the different natural sweeteners, especially steviol glycosides in certain proportion along with other water soluble molecules to provide enhanced sweetness and flavor profile in food and beverage application, which can be blended with other natural caloric sweeteners to impart more desirable sweetness profile. Non-limiting examples of caloric sweeteners include dextrose, fructose, sucrose, maltose, lactose, corn syrup, gluco-syrup derived from different carbohydrates, cane syrup, flavored sugar, honey, molasses, 
     This invention combine the different natural sweeteners, especially steviol glycosides in certain proportion along with other water soluble molecules to provide enhanced sweetness and flavor profile in food and beverage application, which can be blended with other natural non-caloric sweeteners to impart more desirable sweetness profile. Non-limiting examples of natural high intensity sweeteners include steviol glycosides, brazzein, monatin and its salt, neohesperidin dihydrochalcone (NHDC), glycyrrhizic acid and its salts, thaumatin, mogrosides and lu han guo extracts, perillartine, mabinlin, pentadin, miraculin, curculin, neoculin, chlorogenic acid, cynarin, siamenoside and others. 
     This invention combine the different natural sweeteners, especially steviol glycosides in certain proportion along with other water soluble molecules to provide enhanced sweetness and flavor profile in food and beverage application, which can be blended with other synthetic non-caloric sweeteners to impart more desirable sweetness profile. Non-limiting examples of synthetic sweeteners include sucralose, potassium acesulfame, aspartame, alitame, advantame, saccharin, neohesperidin dihydrochalcone synthetic derivatives, cyclamate, neotame, dulcin, suosan, N—[N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-L-α-aspartyl]-phenylalanine 1-methyl ester, N—[N-[3-(3-hydroxy-4-methoxyphenyl)-3-methylbutyl]-α-aspartyl]-L-phenylalanine 1-methyl ester, N—[N-[3-(3-methoxy-4-hydroxyphenyl)propyl]-L-α-aspartyl]-L-phenylalanine 1-methyl ester, salts thereof, and the like. 
     BRIEF SUMMARY OF THE INVENTION 
     The present invention is directed to a taste and flavor modifying composition. The composition includes different steviol glycosides with other water soluble molecules derived from  Stevia  leaf, such as non-limiting examples of plant glycosides, flavonoids, labdane diterpene, triterpenes, which can modify the intensity of a taste and/or a flavor in a food or beverage product. 
     The present invention is also directed to a food or beverage product having an intense taste and flavor profile, wherein the food or beverage product includes a taste and flavor modifying composition comprising the  Stevia  extract of steviol glycosides and water soluble molecules derived from  Stevia  plant. A wide range of food and beverage products, such as, but not limited to, carbonated soft drinks, fruit juices, dairy foods, dairy beverages, baked goods, cereal products, snack foods, and table top sweeteners, may be made in accordance with the present invention. The taste and flavor profile of a food or beverage product including a taste and flavor modifying composition, wherein the taste and flavor modifying composition comprising the  Stevia  extract of steviol glycosides and water soluble molecules derived from  Stevia  plant, may be more intense than a comparative taste and flavor profile of a comparative food or beverage product which does not include the taste and flavor modifying composition. Moreover, the mouthfeel and overall taste perception of a food or beverage product including the taste and flavor modifying composition, wherein the taste and flavor enhancing composition includes the complex mixture of steviol glycosides and water soluble molecules, may be improved in relation to a mouthfeel and overall taste perception of a comparative food or beverage product which does not include the taste and flavor enhancing composition. 
     The present invention is further directed to a method of increasing the taste and flavor intensity 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 modifying composition comprising the  Stevia  extract of steviol glycosides and water soluble molecules derived from  Stevia  plant. The present invention is also directed to a method of improving the organoleptic properties of a food or beverage product including a high fructose syrup, including the step of adding the taste and flavor modifying composition to the food or beverage product. For example, adding the taste and flavor modifying composition may cause the high fructose syrup, such as high fructose corn syrup, to taste more like sugar. Also, if the high fructose syrup is high fructose corn syrup 42 (HFCS 42), adding the taste and flavor enhancing composition may cause the HFCS 42 to taste more like high fructose corn syrup 55 (HFCS 55). 
     The present invention is further directed to a method of increasing the taste and flavor intensity of a medical food and pharma product, including the step of adding a taste and flavor modifying composition to the food or beverage product, wherein the taste and flavor modifying composition comprising the  Stevia  extract of selected steviol glycosides and water soluble molecules derived from  Stevia  plant. The present invention is also directed to a method of improving the organoleptic properties of a medical food or pharma product containing functional food ingredients like vitamins, minerals and amino acids, including the step of adding the taste and flavor modifying composition to the food or beverage product. For example, adding the taste and flavor modifying composition may cause the off-taste due to vitamins, mineral, amino acids and other non-limiting functional ingredients, to improve taste and palatability. 
     The present invention is also directed to a method of making a taste and flavor enhancing composition, including: extracting steviol glycosides and other water soluble molecules from leaves of a  Stevia rebaudiana  plant, and separating the excess steviol glycosides than the amount and type of steviol glycosides required to contribute the taste and flavor modifying characteristics of the  Stevia  extract. 
     The foregoing has outlined rather broadly the features and technical advantages of the present invention in order that the detailed description of the invention that follows may be better understood. Additional features of the invention which form the subject of the claims of the invention will be described hereinafter. It should be appreciated by those skilled in the art that the specific embodiments disclosed may be readily utilized as a basis for modifying or designing other methods or structures for carrying out the same purposes of the present invention. It should also be realized by those skilled in the art that such equivalent constructions do not depart from the spirit and scope of the invention as set forth in the appended claims. The novel features which are believed to be characteristic of the invention, both as to its organization and method of operation, together with further objects and advantages will be better understood from the following description. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  shows the chemical structure of the diterpene glycosides of  Stevia rebaudiana.    
         FIG. 2  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to a cola flavored carbonated soft drink. 
         FIG. 3  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to an iced tea beverage. 
         FIG. 4  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to an iced tea beverage. 
         FIG. 5  is a bar graph showing the effect of  Stevia  extract on the flavor profile of roasted peanuts. 
         FIG. 6  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to tomato ketchup. 
         FIG. 7  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to chocolate milk. 
         FIG. 8  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to poppy seed muffins. 
         FIG. 9  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to tortilla chips. 
         FIG. 10  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to beef jerky. 
         FIG. 11  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to brown gravy. 
         FIG. 12  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to chocolate milk. 
         FIG. 13  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to vanilla custard. 
         FIG. 14  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to chocolate milk. 
         FIG. 15  is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract to vanilla yogurt. 
     
    
    
     DETAILED DESCRIPTION 
     Embodiments of the present invention are described in the following examples. 
     EXAMPLES 
     Example 1A: Detection of Concentration Threshold for Sweetness Recognition 
     To detect the sweetness recognition level of PCS-5001, PCS 1015 and PSB 5005 ( Stevia  extract), the test method outlined by Harman, et al (Food Technology, November 2013) was used with ten trained panelists that have been previously qualified for their taste acuity and trained in the use of a sweetness intensity rating scale. The panelists evaluated a series of aqueous solutions of sucrose and the  Stevia  extract (PCS-5001, PCS-1015, or PSB 5005) at room temperature; the sucrose solutions of 1.5% concentration and the  Stevia  extract solutions with concentrations ranging between 100 and 120 ppm for PCS-5001,70-80 ppm for PCS-1015, and 60-70 ppm for PSB 5005 were prepared with filtered water. The objective of the test was to determine the sweetness recognition level of the  Stevia  extract. The evaluations were done in triplicate using the same panelists so that a total of 30 values were generated for each average data point. 
     The samples were coded and presented in random order to panel members to taste and determine which sample was sweeter (ASTM E2164-08: Standard Method for Directional Difference Test). Panelists were asked to focus only on sweet attribute of those samples and to use warm water and salt solution in order to cleanse the palate between samples. 
     The results were tallied and significance was calculated by SIM 2000 (Sensory Computer System, NJ). Results are presented in Table 4. The overall sweetness of those samples was barely detectable. The 2-AFC shows that 100 ppm PCS-5001, 70 ppm of PCS-1015 and 60 ppm of PSB 5005 solutions were the least sweet samples and were significantly less sweet then the 1.5% sugar control. The sample with 120 ppm PCS-5001 and 80 ppm PCS-1015 were the sweetest samples showing significantly higher sweetness than the 1.5% sugar control (Table 4). The recognition threshold concentration of  STEVIA  EXTRACT (PCS-5001) in water was determined to be 100 ppm. The recognition threshold concentration of  STEVIA  EXTRACT (PCS-1015) in water was determined to be 70 ppm. The sweetness recognition threshold of  STEVIA  EXTRACT (PSB 5005) in water was determined to be 60 ppm. 
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Sweetness perception of Stevia Extract in different concentration 
               
               
                 against 1.5% sugar solution.  
               
            
           
           
               
               
               
               
               
            
               
                 Comparison of sweetness 
                   
                   
                   
                   
               
               
                 perception of STEVIA 
                 Sugar solution 
                 Stevia Extract 
                   
                   
               
               
                 EXTRACT in water 
                 (1.5%) sweeter? 
                 solution sweeter? 
                 P-Value 
                 Significance 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 PCS-5001: 100 ppm, N=30 
                 23 
                 7 
                 0.0052 
                 *** 
               
               
                 PCS-5001: 110 ppm, N=30 
                 20 
                 10 
                 0.0987 
                 **  
               
               
                 PCS-5001: 120 ppm, N=30 
                 9 
                 21 
                 0.0457 
                 *** 
               
               
                 PCS-1015:  70 ppm, N=30 
                 26 
                 4 
                 0.0001 
                 *** 
               
               
                 PCS-1015:  80 ppm, N=30 
                 5 
                 25 
                 0.0003 
                 *** 
               
               
                 PSB-5005:  60 ppm, N=30 
                 24 
                 6 
                 0.0014 
                 *** 
               
               
                 PSB-5005:  70 ppm, N=30 
                 19 
                 11 
                 0.2005 
                 NS  
               
               
                   
               
            
           
         
       
     
     Example 1B: Sweetness Detection of Concentration Threshold for Sweetness Detection 
     The ten panel members evaluated a series of lemon-lime flavored carbonated soft drink (CSD) sweetened with sucrose and  STEVIA  EXTRACT at room temperature; the evaluations were done in triplicate using the same panelists so that at least 30 values were generated for each average data point. The lemon lime flavored carbonated soft drink control sample had 1.5% sucrose concentration and the test sample contained  STEVIA  EXTRACT (PCS-5001) with concentrations at 110 and 120 ppm or  STEVIA  EXTRACT (PCS-1015) with concentrations of 70 and 90 ppm. Other ingredients in the CSD samples were citric acid, lemon-lime flavor, sodium benzoate, potassium citrate and xanthan gum. The objective of the test was to determine the sweetness detection limit of  STEVIA  EXTRACT. Tests were conducted as outlined in Example 1A. 
     The samples with 120 ppm PCS-5001( STEVIA  EXTRACT) and 90 ppm PCS-1015( STEVIA  EXTRACT) showed no significant difference in sweetness than the 1.5% sugar control. The recognition threshold concentration of PCS-5001( STEVIA  EXTRACT) in a lemon-lime flavored carbonated soft drink water was determined to be 110 ppm. The recognition threshold concentration of PCS-1015 ( STEVIA  EXTRACT) in a lemon-lime flavored carbonated soft drink water was determined to be 70 ppm. Results are shown in table 5. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Sweetness perception of STEVIA EXTRACT in different concentrations against 
               
               
                 1.5% sugar solution in a typical carbonated soft drink (CSD) 
               
            
           
           
               
               
               
               
               
            
               
                 Sweetness perception of  
                 CSD sample with 
                 CSD sample with 
                   
                   
               
               
                 SIEVIA EXTACT in CSD 
                 Sugar sweeter? 
                 Stevia Sweeter? 
                 P-Value 
                 Significance 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 PCS-5001: 110 ppm, N=30 
                 23 
                 7 
                 0.0052 
                 *** 
               
               
                 PCS-5001: 120 ppm, N=36 
                 20 
                 16 
                 0.677 
                 NS 
               
               
                 PCS-1015:  70 ppm, N=30 
                 21 
                 9 
                 0.0428 
                 *** 
               
               
                 PCS-1015:  90 ppm, N=30 
                 12 
                 18 
                 0.3616 
                 NS 
               
               
                   
               
            
           
         
       
     
     Example 2: Effect of  Stevia  Extract on Flavor Modification in a Typical Carbonated Soft Drink Application 
     A cola flavored carbonated soft drink was developed to evaluate the effect of PCS-5001 and PCS-1015 ( Stevia  extract) on the sweetness and flavor profile of the beverage that was sweetened with sugar and  Stevia  sweetener to achieve 30% sugar reduction (Table 6). The samples with and without PCS-5001 or PCS-1015 were evaluated by thirty consumer panel members, who assigned relative values to each sample for overall Liking, sweetness, vanilla flavor, brown note, and aftertaste on a 10-pt continuous intensity scale as outlined in Table 7. 
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Cola flavored Soft drink for sensory evaluation 
               
            
           
           
               
               
               
               
            
               
                   
                 Control: 
                 Test: 30% Sugar 
                 Test: 30% Sugar 
               
               
                 COLA BEVERAGE 
                 30% 
                 Reduction with 
                 Reduction with 
               
               
                 FORMULA 
                 Sugar Reduction 
                 PCS-5001 
                 PCS-1015 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Water 
                 91.68 
                 91.67 
                 91.67 
               
               
                 Sugar 
                 7.89 
                 7.89 
                 7.89 
               
               
                 Cola Flavor-Flavor Systems 
                 0.375 
                 0.375 
                 0.375 
               
               
                 Phosphoric Acid 85% 
                 0.0333 
                 0.0333 
                 0.0333 
               
               
                 Caffeine 
                 0.0100 
                 0.0100 
                 0.0100 
               
               
                 Steviol glycoside 
                 0.0100 
                 0.0100 
                 0.0100 
               
               
                 PCS-5001 
                   
                 0.0110 
                   
               
               
                 PCS-1015 
                   
                   
                 0.0080 
               
               
                 Total 
                 100 
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
               
                   
               
               
                 Sensory evaluation of Cola flavored carbonated soft drink 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 Nature of Participants: 
                 Company employees 
               
               
                 Number of Sessions 
                 1 
               
               
                 Number of 
                 30 
               
               
                 Participants: 
                   
               
               
                 Test Design: 
                 Balanced, randomized within pair. Blind 
               
               
                 Sensory Test 
                 Intensity and acceptance ratings 
               
               
                 Method: 
                   
               
               
                 Environmental 
                 Standard booth lighting 
               
               
                 Condition 
                   
               
               
                 Attributes and Scales: 
                   
               
            
           
           
               
            
               
                 Overall Acceptance on a 10-pt hedonic scale where 10 = Extremely Like and 0 =  
               
               
                 Extremely Dislike 
               
               
                 Overall Liking, Sweetness, Vanilla flavor, Brown note, and Sweet Aftertaste. 
               
               
                 10-pt continuous intensity scale where 0 = Imperceptible and 10 = Extremely 
               
            
           
           
               
               
            
               
                 Pronounced 
                   
               
               
                 Statistical Analysis: 
                 ANOVA (by Block) with Post Hoc Duncan&#39;s Test 
               
               
                 Sample Size 
                 ~1.5 oz in a clear capped plastic cup 
               
               
                 Serving Temperature 
                 Refrigerated temperature (~45° F.) 
               
               
                 Serving/Panelists 
                 Samples served simultaneously. Panelists instructed to 
               
               
                 Instruction: 
                 read ingredient statement, evaluate each sample. 
               
               
                   
               
            
           
         
       
     
       FIG. 2  shows the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract (PCS-5001). The results indicated the sample containing  Stevia  extract PCS-5001 and the sample containing PCS-1015 had significantly higher cola flavor, vanilla flavor, brown spice notes and overall liking compared to the control samples (at 95% confidence). The sample containing PCS-5001 had directionally lower bitterness, and bitter aftertaste intensity compared to the control samples (at 90% and 95% confidence respectively). The sample containing PCS-1015 had directionally lower bitterness, and sweet aftertaste intensity compared to the control samples (at 80% confidence). In addition, the sample with  Stevia  extract (PCS-1015) had significantly lower bitter aftertaste compared to the control sample (at 95% confidence). 
     Example 3: Peach Flavored Tea Beverage for Sensory Evaluation 
     A peach flavored black tea drink was developed to evaluate the effect of  STEVIA  EXTRACT on the sweetness and flavor profile of the beverage that was sweetened with sugar and  Stevia  sweetener to achieve 30% sugar reduction (Table 8). The samples with and without  STEVIA  EXTRACT were evaluated as outlined in EXAMPLE 2 by thirty consumer panel members, who assigned relative values to sweetness, bitterness, peach flavor, tea flavor, acid intensity, astringency, and aftertaste on 10-pt continuous intensity scale where 0=Imperceptible and 10=extremely pronounced. 
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Peach Flavored Tea Beverage samples for sensory evaluation 
               
            
           
           
               
               
               
               
            
               
                   
                   
                 Reduce Sugar 
                 Reduce Sugar 
               
               
                   
                 Reduced Sugar 
                 Tea with PCS- 
                 Tea with PCS- 
               
               
                   
                 Tea 
                 5001 
                 1015 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Water 
                 95.71 
                 95.70 
                 95.71 
               
               
                 Sucrose 
                 3.850 
                 3.850 
                 3.850 
               
               
                 Black Tea Powder 
                 0.275 
                 0.275 
                 0.275 
               
               
                 Citric Acid 
                 0.0880 
                 0.0880 
                 0.0880 
               
               
                 Peach Flavor 
                 0.0330 
                 0.0330 
                 0.0330 
               
               
                 Sodium Citrate 
                 0.0150 
                 0.0150 
                 0.0150 
               
               
                 Potassium Sorbate 
                 0.0150 
                 0.0150 
                 0.0150 
               
               
                 Steviol Glycoside 
                 0.0140 
                 0.0140 
                 0.0140 
               
               
                 Stevia Extract PCS-5001 
                   
                 0.0120 
                   
               
               
                 Stevia Extract PCS-1015 
                   
                   
                 0.0080 
               
               
                 Xanthan Gum-TIC 
                 0.0013 
                 0.0013 
                 0.0013 
               
               
                   
               
            
           
         
       
     
       FIG. 3  shows the modification of flavor and sweetness profiles contributed by the addition of  STEVIA  EXTRACT (PCS-5001) in peach flavored ice tea beverage. The results indicated that the test sample containing PCS-5001 had significantly higher peach flavor, and overall liking (at 95%, confidence). The sample containing PCS-5001 had significantly lower astringency than the control sample (at 95% confidence). The results shown in  FIG. 4  indicated that the test sample containing PCS-1015 had significantly higher peach flavor, black tea flavor, and overall liking (at 95%, confidence). The sample PCS-1015 also had significantly lower astringency, sweet intensity, bitter intensity, and bitter aftertaste than the Control sample (at 95% confidence). In addition, the PCS-1015 sample had lower sweet aftertaste intensity than the Control sample at 90% confidence). 
     Example 4: Effect of  Stevia  Extract on Flavor Modification of Savory Applications 
     A seasoning blend was developed to determine the flavor modification effect of  Stevia  extract in a seasoning blend on reduced sugar roasted peanut samples. Thirty consumer panel members evaluated two samples of the peanuts for overall acceptance and attribute intensities (overall flavor, saltiness, sweetness, smoke flavor, spice/heat intensity, peanut flavor, chili powder flavor, bitterness and lingering sweet aftertaste intensity). The two samples (Table 9) included: 1) 50% sugar reduced control sample containing  Stevia  glycosides, and 2) 50% reduced sugar test sample containing steviol glycoside and  Stevia  extract, PCS-5001 or PCS-1015. 
     The objective of the test was to determine if the addition of  Stevia  extract affects the flavor profile of a savory snack food. The results indicated that the addition of PCS-5001 at 110 ppm and PCS-1015 at 70 ppm provided flavor modification ( FIG. 5 ). The test samples containing 110 ppm PCS-5001 had significantly higher salt intensity, smoke flavor, and bitter intensity compared to the control (95% confidence). The test sample also had lower sweet intensity than the control (95% confidence). In addition, the test sample containing  Stevia  extract had directionally higher spice and chili notes (90% confidence). The test sample containing PCS-1015 had significantly higher salt intensity than the control sample (at 95% confidence). The test sample showed an increase in heat/spice intensity, and chili flavor compared to the control. 
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Effect of STEVIA EXTRACT on snack and seasoning applications 
               
            
           
           
               
               
               
               
            
               
                   
                 Steviol 
                 Steviol Glycoside + 
                 Steviol Glycoside + 
               
               
                   
                 Glycoside 
                 Stevia Extract 
                 Stevia Extract 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 Unsalted Peanuts 
                 86.8 
                 86.8 
                 86.8 
               
               
                 Vegetable oil 
                 2.93 
                 2.93 
                 2.93 
               
               
                 Sugar 
                 5.88 
                 5.88 
                 5.88 
               
               
                 Salt 
                 2.93 
                 2.93 
                 2.93 
               
               
                 Chilli powder 
                 0.174 
                 0.174 
                 0.174 
               
               
                 Cumin powder 
                 0.286 
                 0.286 
                 0.286 
               
               
                 Garlic powder 
                 0.156 
                 0.156 
                 0.156 
               
               
                 Cayenne pepper 
                 0.156 
                 0.156 
                 0.156 
               
               
                 Smoke liquid 
                 0.729 
                 0.729 
                 0.729 
               
               
                 Steviol Glycoside 
                 0.0243 
                 0.0243 
                 0.0243 
               
               
                 PCS-5001 
                   
                 0.0110 
                   
               
               
                 PCS-1015 
                   
                   
                 0.0070 
               
               
                 Total wt. (g) 
                 100 
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 10 
               
               
                   
               
               
                 Sensory evaluation of snack and seasoning applications 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 Nature of Participants: 
                 Company employees 
               
               
                 Number of Sessions 
                 1 
               
               
                 Number of Participants: 
                 30 
               
               
                 Test Design: 
                 Balanced, randomized within pair. Blind 
               
               
                 Sensory Test Method: 
                 Intensity and acceptance ratings 
               
               
                 Environmental Condition 
                 Standard booth lighting 
               
               
                 Attributes and Scales: 
                   
               
            
           
           
               
            
               
                 Overall Acceptance on a 9-pt hedonic scale where 9 = Like Extremely, 5 = Neither Like 
               
               
                 Nor Dislike, and 1 = Dislike Extremely 
               
               
                 Overall Flavor, Saltiness, Sweetness, Smoke Intensity, Heat/spice intensity, peanut 
               
               
                 flavor, chili powder and Aftertaste Intensity (sweet and bitter) on a 10-pt continuous 
               
               
                 intensity scale where 0 = Imperceptible and 10 = Extremely Pronounced 
               
               
                 Open Ended General Comments 
               
            
           
           
               
               
            
               
                 Statistical Analysis: 
                 ANOVA (by Block) with Post Hoc Duncan&#39;s Test 
               
               
                 Sample Size 
                 ~1.5 oz in a clear capped plastic cup 
               
               
                 Serving Temperature 
                 Room temperature (~70° F.) 
               
               
                 Serving/Panelists 
                 Samples served simultaneously. Panelists evaluate each 
               
               
                 Instruction: 
                 sample once. 
               
               
                   
               
            
           
         
       
     
     Example 5: Flavor Modification of Sauce and Vegetable Preparation 
     A tomato ketchup preparation was developed to determine the flavor modification effect of  Stevia  extract (PCS-1015). A panel of thirty company employees evaluated the overall acceptance and attribute intensities (tomato, onion, vinegar, sweet, saltiness, bitterness and aftertaste) of each sample. The sensory evaluation methodology outlined in Example 4 was adopted for the sauce samples as presented in Table 11. 
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Effect of PCS-1015 (stevia extract) on tomato ketchup 
               
            
           
           
               
               
               
            
               
                   
                   
                 Steviol Glycoside 
               
               
                   
                 Steviol Glycoside 
                 w/Stevia Extract 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Tomato Juice (Sieved) 
                 52.4863 
                 52.4793 
               
               
                 Tomato Puree 
                 24.6236 
                 24.6236 
               
               
                 White Distilled Vinegar 
                 11.3454 
                 11.3454 
               
               
                 Water 
                 1.5845 
                 1.5845 
               
               
                 Sucrose 
                 2.6511 
                 2.6511 
               
               
                 Tomato Paste 
                 5.8311 
                 5.8311 
               
               
                 Onion Powder 
                 0.8649 
                 0.8649 
               
               
                 Salt 
                 0.5811 
                 0.5811 
               
               
                 Steviol glycoside 
                 0.032 
                 0.032 
               
               
                 Stevia Extract (PCS 1015) 
                   
                 0.007 
               
               
                 Total 
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
       FIG. 6  shows the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract (PCS-1015). The results indicate the test samples containing  Stevia  extract, PCS-1015, had a significant increase in herbal notes, and savory (onion/garlic) notes at a 95% confidence interval. The test sample containing PCS-1015 had directionally lower bitterness, bitter aftertaste and overall liking at a 90% confidence interval compared to the control sample. 
     Example 6: Effect of PCS-1015 ( Stevia  Extract) on Flavor Modification of Dairy Applications 
     A chocolate flavored dairy beverage was developed to determine the flavor modification effect of  Stevia  extract (PCS-1015) in dairy beverage. The panel evaluated samples of chocolate milk for overall acceptance and attribute intensities (chocolate flavor, dairy notes, sweetness, bitterness and aftertaste). The two samples (Table 12) included: 1) 50% sugar reduced control sample containing  Stevia  glycosides, and 2) 50% reduced sugar test sample containing  Stevia  glycoside and 80 ppm of  Stevia  extract, PCS-1015. 
     
       
         
           
               
             
               
                 TABLE 12 
               
             
            
               
                   
               
               
                 Effect of PCS-1015 (stevia extract) on flavored dairy beverage 
               
            
           
           
               
               
               
            
               
                   
                 50% Total Sugar 
                 50% Total Sugar 
               
               
                   
                 Reduction  
                 Reduction with stevia  
               
               
                   
                 with steviol 
                 extract and stevia 
               
               
                 Dairy Formula 
                 glycoside 
                 glycoside 
               
               
                   
               
            
           
           
               
               
               
            
               
                 2% Reduced fat Milk 
                 96.5803 
                 96.5753 
               
               
                 Sugar 
                 2.40 
                 2.40 
               
               
                 Cocoa Powder 
                 0.80 
                 0.80 
               
               
                 Palsgaard 150 ChoMilk 
                 0.20 
                 0.20 
               
               
                 Steviol Glycosides 
                 0.0197 
                 0.0197 
               
               
                 PCS-1015 
                   
                 0.080 
               
               
                 Total 
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 13 
               
               
                   
               
               
                 Sensory evaluation of Dairy beverage 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 Nature of Participants: 
                 Company employees 
               
               
                 Number of Sessions 
                 1 
               
               
                 Number of Participants: 
                 30 
               
               
                 Test Design: 
                 Balanced, randomized within pair. Blind 
               
               
                 Sensory Test Method: 
                 Intensity and acceptance ratings 
               
               
                 Environmental 
                 Standard booth lighting 
               
               
                 Condition 
                   
               
               
                 Attributes and Scales: 
                   
               
            
           
           
               
            
               
                 Overall Acceptance on a 10-pt hedonic scale where 10 = Extremely Like and 0 = 
               
               
                 Extremely Dislike 
               
               
                 Overall Liking, sweetness, bitterness, dairy notes, chocolate, and Aftertaste. 10- 
               
               
                 pt continuous intensity scale where 0 = Imperceptible and 10 = Extremely 
               
               
                 Pronounced 
               
            
           
           
               
               
            
               
                 Statistical Analysis: 
                 ANOVA (by Block) with Post Hoc Duncan&#39;s Test 
               
               
                 Sample Size 
                 ~1.5 oz. in a clear capped plastic cup 
               
               
                 Serving Temperature 
                 Refrigerated temperature (~45° F.) 
               
               
                 Serving/Panelists 
                 Samples served simultaneously. Panelists instructed to read 
               
               
                 Instruction: 
                 ingredient statement, evaluate each sample. 
               
               
                   
               
            
           
         
       
     
       FIG. 7  shows the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract (PCS-1015). The results indicate the 50% sugar reduced sample containing steviol glycoside sweetener and  Stevia  extract, PCS-1015, had significantly higher chocolate flavor. 
     Example 7: Effect of  Stevia  Extract (PCS-5001) on Flavor Modification of Baked Goods Applications 
     A lemon poppy seed flavored muffin formulation was developed to determine the flavor modification effect of  Stevia  extract (PCS-5001) in baked good applications. To test the contribution of PCS-5001 in baked goods, lemon flavored poppy seed muffins were baked with a 45% sugar reduced formulation with steviol glycoside as control, and sugar reduced formulation with steviol glycoside and  Stevia  extract (PCS-5001) as a test sample as shown in Table 14. A thirty member consumer panel evaluated two samples of lemon poppy seed muffins for several attributes (lemon, vanilla flavors, brown notes, sweet &amp; bitter aftertaste). 
     
       
         
           
               
             
               
                 TABLE 14 
               
             
            
               
                   
               
               
                 Effect of PCS-5001 (stevia extract) on baked goods 
               
            
           
           
               
               
               
            
               
                   
                 Steviol  
                 Steviol  
               
               
                   
                 Glycoside 
                 glycoside w/ 
               
               
                   
                 (400 ppm)  
                 120 ppm  
               
               
                 Ingredients 
                 Control 
                 stevia extract 
               
               
                   
               
            
           
           
               
               
               
            
               
                 DRY Ingredients 
                   
                   
               
               
                 Sucrose 
                 12.3722 
                 12.3682 
               
               
                 All Purpose Flour 
                 17.6434 
                 17.6434 
               
               
                 Whole Wheat Flour 
                 5.8763 
                 5.8763 
               
               
                 Poppy Seeds 
                 1.0648 
                 1.0648 
               
               
                 Maltodextrin—10DE 
                 2.1368 
                 2.1368 
               
               
                 Fibersol2 (ADM/Matsutani) 
                 1.0648 
                 1.0648 
               
               
                 Modified Starch—Inscosity 656 
                 1.0648 
                 1.0648 
               
               
                 Lemon Flavor—Firmenich 
                 0.8860 
                 0.8860 
               
               
                 Salt (Sodium Chloride) 
                 0.7479 
                 0.7479 
               
               
                 Baking Powder 
                 1.0648 
                 1.0648 
               
               
                 Baking Soda 
                 0.3205 
                 0.3205 
               
               
                 Steviol Glycoside 
                 0.0400 
                 0.0400 
               
               
                 Stevia extract (PCS-5001) 
                   
                 0.0120 
               
               
                 Wet Ingredients 
                   
                   
               
               
                 Milk, 2% 
                 27.2444 
                 27.2444 
               
               
                 Soybean Oil 
                 11.7525 
                 11.7525 
               
               
                 Whole Eggs 
                 8.5473 
                 8.5473 
               
               
                 Water 
                 5.3420 
                 5.3420 
               
               
                 Yogurt, Plain Nonfat 
                 1.6026 
                 1.6026 
               
               
                 Lemon Juice, 100% 
                 0.6410 
                 0.6410 
               
               
                 Vanilla Extract 
                 0.5342 
                 0.5342 
               
               
                   
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
       FIG. 8  shows the modification of flavor and sweetness profiles caused by the addition of  Stevia  extract (PCS-5001). The panel found that the addition of  Stevia  extract provided an increase in brown note than control sample without  Stevia  extract (at 90% confidence). 
     Example 8: Effect of  Stevia  Extract (PCS-5001) on Flavor Modification of Reduced Sodium Applications 
     A 30% salt reduced tortilla chip formulation was developed to determine the flavor modification effect of  Stevia  extract (PCS-5001) in a salt reduced applications. To test the contribution of PCS-5001 in a salt reduced application, cheddar cheese flavor tortilla chips were coated with a control salt formulation, and a 30% salt reduced formulation with  Stevia  extract (PCS-5001) as a test sample as shown in Table 15. A sixteen member consumer panel evaluated two samples of cheddar cheese flavored tortilla chips for different attributes (sweet intensity, saltiness, cheese flavor, dairy notes, corn flavor, bitterness, and sweet &amp; bitter aftertaste). 
       FIG. 9  shows the modification of flavor and salt perception caused by the addition of  Stevia  extract (PCS-5001). The panel found the addition of  Stevia  extract in a 30% salt reduced formulation provided an increase in salt perception, parity to the full sodium control. In addition,  Stevia  extract provided an increase in sweet intensity and dairy note higher than control sample without  Stevia  extract (at 95% confidence). 
     
       
         
           
               
             
               
                 TABLE 15 
               
             
            
               
                   
               
               
                 Tortilla Chips with Cheddar Cheese 30% less sodium 
               
            
           
           
               
               
               
            
               
                   
                 Control 
                 30% Less Salt 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Corn chips 
                 78 
                 78.33 
               
               
                 Cheese seasoning 
                 10 
                 10.04 
               
               
                 Vegetable Oil 
                 11 
                 11.05 
               
               
                 Added Salt 
                 1 
                 0.57 
               
               
                 PCS-5001 
                   
                 0.01 
               
               
                 Total w (g) 
                 100 
                 100.00 
               
               
                   
               
            
           
         
       
     
     Example 9: Effect of  Stevia  Extract (PCS-5001) on Flavor Modification of Dried Meat Applications 
     A beef jerky formulation was developed to determine the flavor modification effect of  Stevia  extract (PCS-5001) in a dried meat applications. To test the contribution of PCS-5001 in a dried meat application, flank steak was marinated with a reduced sugar control formulation, and a 30% sugar reduced formulation with steviol glycosides and  Stevia  extract (PCS-5001) as a test sample as shown in Table 16. A twenty member consumer panel evaluated two samples of beef jerky for different attributes (sweet intensity, saltiness, black pepper, teriyaki flavor, fat-like intensity, beef flavor and sweet aftertaste). 
       FIG. 10  shows the modification of flavor and salt perception caused by the addition of  Stevia  extract (PCS-5001). The panel found the addition of  Stevia  extract in a 30% sugar reduced formulation provided an increase in salt perception. 
     
       
         
           
               
             
               
                 TABLE 16 
               
             
            
               
                   
               
               
                 30% sugar reduced Beef Jerky 
               
            
           
           
               
               
               
            
               
                   
                 Control (%) 
                 Stevia Extract 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Flank Steak 
                 75.44 
                 75.44 
               
               
                 Balsamic vinegar 
                 10.15 
                 10.15 
               
               
                 Salt 
                 2.46 
                 2.46 
               
               
                 Pepper 
                 0.83 
                 0.83 
               
               
                 Sugar 
                 6.88 
                 6.88 
               
               
                 Liquid smoke 
                 0.86 
                 0.86 
               
               
                 Water 
                   
                   
               
               
                 Garlic powder 
                 0.44 
                 0.44 
               
               
                 Onion powder 
                 0.44 
                 0.44 
               
               
                 Steviol Glycoside 
                 0.018 
                 0.018 
               
               
                 PCS-5001 (stevia extract) 
                   
                 0.0100 
               
               
                 Worcestershire sauce 
                 2.46 
                 2.46 
               
               
                   
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
     Example 10: Effect of  Stevia  Extract (PCS-5001) on Flavor Modification of Reduced Sodium Applications in Brown Gravy 
     A 30% sodium reduced brown gravy formulation was developed to determine the flavor modification effect of  Stevia  extract (PCS-5001) in a salt reduced applications. To test the contribution of PCS-5001 in a salt reduced application, a 30% sodium reduced brown gravy formulation, and a 30% salt reduced formulation with  Stevia  extract (PCS-5001) as a test sample. A thirty member consumer panel evaluated two samples of brown gravy for different attributes (sweet intensity, saltiness, black pepper, beef flavor, and onion/savory notes, bitterness, and sweet &amp; bitter aftertaste). 
       FIG. 11  shows the modification of flavor and salt perception caused by the addition of  Stevia  extract (PCS-5001). The panel found the addition of  Stevia  extract in a 30% salt reduced formulation provided an increase in salt perception compared to 30% sodium reduced control. In addition,  Stevia  extract provided an increase in savory and black pepper note higher than control sample without  Stevia  extract (at 95% confidence). There was also a decrease in bitter aftertaste. 
     Example 11: Effect of  Stevia  Extract on Flavor Modification of Dairy Product 
     To evaluate the contribution of PCS-1015 (MLD-1), a  Stevia  extract, to a dairy product, two 50% reduced sugar chocolate milk samples were prepared and tested by a consumer panel of 30 company employees. The consumer panel evaluated those two samples of chocolate milk for overall acceptance and attribute intensities (chocolate flavor, dairy notes, sweetness, bitterness and aftertaste) in two sessions. In session one, the two samples included: 1) a 50% sugar reduced control sample containing PureCircle Alpha (steviol glycoside sweetener) and 2) 50% sugar reduced test sample containing PureCircle Alpha and 70 ppm PCS-1015 (MLD-1). In session two, the two samples included: 1) a 50% sugar reduced control sample containing PureCircle Alpha (steviol glycoside sweetener) and 2) 50% sugar reduced test sample containing PureCircle Alpha and 80 ppm PCS-1015 (MLD-1). Tables 17 shows the formula of the control and test samples of 50% reduced sugar. 
     
       
         
           
               
             
               
                 TABLE 17 
               
             
            
               
                   
               
               
                 50% sugar reduced Chocolate Milk with PCS-1015 
               
            
           
           
               
               
               
               
            
               
                   
                   
                 50% Total  
                 50% Total  
               
               
                   
                 50% Total  
                 Sugar 
                 Sugar 
               
               
                   
                 Sugar  
                 Reduction  
                 Reduction  
               
               
                   
                 Reduction  
                 with 
                 with  
               
               
                   
                 with  
                 PC  
                 PC 
               
               
                   
                 PureCircle 
                 Alpha &amp; 
                 Alpha &amp;  
               
               
                 Dairy Formula 
                 Alpha 
                 PCS-1015 
                 PCS-1015 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 2% Reduced  
                 96.5803 
                 96.5743 
                 96.5753 
               
               
                 fat Milk 
                   
                   
                   
               
               
                 Sugar 
                 2.40 
                 2.40 
                 2.40 
               
               
                 Cocoa Powder  
                 0.80 
                 0.80 
                 0.80 
               
               
                 10/12 
                   
                   
                   
               
               
                 Palsgaard 150  
                 0.20 
                 0.20 
                 0.20 
               
               
                 ChoMilk 
                   
                   
                   
               
               
                 PureCircle  
                 0.0197 
                 0.0197 
                 0.0197 
               
               
                 Alpha 
                   
                   
                   
               
               
                 PCS-1015  
                   
                 0.0070 
                 0.0080 
               
               
                 (MLD-1) 
                   
                   
                   
               
               
                 Total 
                 100 
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
     Table 18 shows the sensory results with the two test samples. Both test samples showed the impact of the  Stevia  extract (PCS 1015) on the Chocolate flavor notes and dairy note. At 80 ppm use level, the chocolate milk sample showed better sweetness profile and overall liking than the control sample.  FIG. 12  shows the comparison of the taste profile between the control and the test sample with 80 ppm  Stevia  extract PCS 1015. 
     
       
         
           
               
             
               
                 TABLE 18 
               
               
                   
               
               
                 Summary of the overall acceptance and mean attribute 
               
               
                 intensity results for each reduced sugar chocolate  
               
               
                 milk samples tested by 30 panel members. 
               
               
                   
               
             
            
               
                 Summary of Mean-Scores, P-Values, and Significance 
               
               
                 Test Result Code—chocolate milk with 70 ppm MLD-1 
               
            
           
           
               
               
               
               
               
            
               
                   
                 197 ppm  
                 70 ppm  
                   
                   
               
               
                   
                 of 
                 of 
                   
                   
               
               
                   
                 Alpha 
                 MLD-1  
                   
                   
               
               
                   
                 Only  
                 w/ PC 
                   
                   
               
               
                 Attribute 
                 (Control) 
                 Alpha 
                 P-Value 
                 Sig 
               
               
                   
               
               
                 Sweet Intensity 
                 8.85 
                 8.89 
                 0.8555 
                 NS 
               
               
                 Chocolate Flavor 
                 6.82 b 
                 7.70 a 
                 0.0482 
                 *** 
               
               
                 Dairy Note 
                 3.61 b 
                 4.19 a 
                 0.1934 
                 * 
               
               
                 Bitterness 
                 0.84 
                 0.83 
                 0.9500 
                 NS 
               
               
                 Bitter Aftertaste 
                 0.74 
                 0.70 
                 0.6096 
                 NS 
               
               
                 Sweet Aftertaste 
                 3.02 
                 3.15 
                 0.7232 
                 NS 
               
               
                 Overall Liking 
                 7.12 
                 7.42 
                 0.5114 
                 NS 
               
               
                   
               
            
           
           
               
            
               
                 Summary of Mean-Scores, P-Values, and Significance 
               
               
                 Test Result Code—chocolate milk with 80 ppm MLD1 
               
            
           
           
               
               
               
               
               
            
               
                   
                 197 ppm  
                 80 ppm  
                   
                   
               
               
                   
                 of 
                 of 
                   
                   
               
               
                   
                 Alpha 
                 MLD-1  
                   
                   
               
               
                   
                 only  
                 w/ PC 
                   
                   
               
               
                 Attribute 
                 (Control) 
                 Alpha 
                 P-Value 
                 Sig 
               
               
                   
               
               
                 Sweet Intensity 
                 8.90 b 
                 9.05 a 
                 0.1557 
                 * 
               
               
                 Chocolate Flavor 
                 6.89 b 
                 7.53 a 
                 0.0048 
                 *** 
               
               
                 Dairy Note 
                 4.12 b 
                 4.44 a 
                 0.1470 
                 * 
               
               
                 Bitterness 
                 0.49 
                 0.35 
                 0.2473 
                 NS 
               
               
                 Bitter Aftertaste 
                 0.71 a 
                 0.55 b 
                 0.1824 
                 * 
               
               
                 Sweet Aftertaste 
                 2.66 
                 2.82 
                 0.5177 
                 NS 
               
               
                 Overall Liking 
                 6.49 b 
                 6.89 a 
                 0.1908 
                 * 
               
               
                   
               
               
                 * = 80% CI, ** = 90% CI, *** = 95% CI 
               
            
           
         
       
     
     Example 12: Effect of  Stevia  Extract on Desserts (Vanilla Custard) 
     To test the contribution of the  Stevia  extract, PCS-1015 in gelatin and puddings, two 30% calorie reduced vanilla custard samples were tested: 1) sweetened with PureCircle Alpha, a PureCircle  Stevia  sweetener, 2) sweetened with PureCircle Alpha and PCS-1015 (MLD-1). Table 19 shows the formulation of the control and test samples. A panel of 30 trained panelists with extensive experience in profiling sensory attributes tasted both samples. 
     To prepare the sample, blend the PureCircle Alpha and the test ingredient (PCS-1015) with the dry ingredients. Add the dry ingredients to the milk using good agitation. Heat on low until all ingredients are dissolved. Heat up to 95° C. for 10 minutes to cook up the starches. Add flavors, stir it, cool, stir it before place it in the refrigerator. Serve at chilled in 1 oz cups. 
     
       
         
           
               
             
               
                 TABLE 19 
               
             
            
               
                   
               
               
                 Reduced sugar dessert (Vanilla Custard) with PCS-1015 
               
            
           
           
               
               
               
            
               
                   
                 Control with 
                 Test with PureCircle 
               
               
                   
                 PureCircle Alpha 
                 Alpha w/ stevia extract 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Milk ( 1% fat) 
                 94.27 
                 94.27 
               
               
                 Sucrose 
                 4.00 
                 4.00 
               
               
                 Starch Perma Flo  
                 1.25 
                 1.25 
               
               
                 Tate &amp; Lyle 
                   
                   
               
               
                 TIC Carrageenan 
                 0.09 
                 0.09 
               
               
                 Salt 
                 0.06 
                 0.06 
               
               
                 ROHA Beta Carotene 
                 0.05 
                 0.05 
               
               
                 French Vanilla Flavor 
                 0.15 
                 0.15 
               
               
                 UV 420-066-7 
                   
                   
               
               
                 Steviol Glycoside 
                 0.0166 
                 0.0166 
               
               
                 Stevia Extract 
                 — 
                 0.0080 
               
               
                 Total 
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
     The trained panel found that the test sample had stronger sweet intensity, vanilla, dairy flavor notes and overall liking at 80% confidence. The sample containing  Stevia  extract also had significantly higher egg note at 95% confidence.  FIG. 13  shows the pictorial rendition of the sensory difference between the control and test dessert samples 
     
       
         
           
               
             
               
                 TABLE 20 
               
             
            
               
                   
               
               
                 Summary of the overall acceptance and mean attribute  
               
               
                 intensity results for reduced sugar dessert  
               
               
                 (Vanilla Custard) with PCS-1015 
               
            
           
           
               
               
               
               
               
            
               
                   
                 166 ppm  
                   
                   
                   
               
               
                   
                 of  
                 70 ppm  
                   
                   
               
               
                   
                 Alpha 
                 of  
                   
                   
               
               
                   
                 Only  
                 MLD-1 
                   
                   
               
               
                 Attribute 
                 (Control) 
                 with Alpha 
                 P-Value 
                 Sig 
               
               
                   
               
               
                 Sweet Intensity 
                 7.01 a 
                 7.13 b 
                 0.1095 
                 * 
               
               
                 Vanilla Flavor 
                 3.22 a 
                 3.5 b 
                 0.1299 
                 * 
               
               
                 Egg Note 
                 1.22 a 
                 1.56 b 
                 0.0497 
                 *** 
               
               
                 Dairy / Creaminess 
                 3.04 a 
                 3.22 b 
                 0.1164 
                 * 
               
               
                 Bitterness 
                 0.43 
                 0.5 
                 0.3001 
                 NS 
               
               
                 Bitter Aftertaste 
                 0.36 
                 0.38 
                 0.7692 
                 NS 
               
               
                 Sweet Aftertaste 
                 2.23 
                 2.24 
                 0.8794 
                 NS 
               
               
                 Overall Liking 
                 6.49 a 
                 6.87 b 
                 0.1149 
                 * 
               
               
                   
               
            
           
         
       
     
     Example 13: Effect of  Stevia  Extract on Flavor Modification of Chocolate-Flavored Beverage with Cocoa Powder Reduction 
     A chocolate flavored dairy beverage was developed to determine the flavor modification effect of  Stevia  extract flavor with modifying properties (FMP) in a dairy beverage. The two samples included: 1) control sample with full amounts of sugar and cocoa powder, and 2) test sample with 15% reduced sugar and 20% reduced cocoa, containing 60 ppm of  Stevia  extract FMP, as shown in Table 21. 
     
       
         
           
               
             
               
                 TABLE 21 
               
             
            
               
                   
               
               
                 Reduced Sugar and Cocoa Chocolate-Flavored Beverage  
               
               
                 (Chocolate Milk) with Stevia Extract 
               
            
           
           
               
               
               
            
               
                   
                   
                 Test with  
               
               
                   
                 Control with  
                 Reduced 
               
               
                   
                 Full Amount  
                 Sugar and  
               
               
                   
                 of Sugar 
                 Cocoa with  
               
               
                   
                 and Cocoa 
                 Stevia Extract 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Milk, 2% milkfat 
                 85.200 
                 86.554 
               
               
                 Sugar 
                 8.00 
                 6.80 
               
               
                 Hot Water, 190° F. 
                 6 
                 6 
               
               
                 Natural 10/12  
                 0.80 
                 0.64 
               
               
                 Cocoa Powder 
                   
                   
               
               
                 Stevia Extract  
                 — 
                 0.006 
               
               
                 FMP (PSB-5005) 
                   
                   
               
               
                 Total 
                 100 
                 100 
               
               
                   
               
            
           
         
       
     
     A 15 member trained panel evaluated samples of chocolate milk for overall acceptance and attribute intensities (sweet intensity, bitterness, cocoa flavor, dairy note, sweet aftertaste and bitter aftertaste). The parameters for the sensory evaluation are shown in Table 22. 
     Table 23 shows the sensory results for the control and test products. The test product with 20% reduced cocoa powder and  Stevia  extract FMP shows no significant difference in cocoa flavor from the control. The test sample with 60 ppm  Stevia  extract FMP was higher in sweet intensity (90% confidence) and sweet aftertaste (directional).  FIG. 14  illustrates this comparison. 
     
       
         
           
               
             
               
                 TABLE 22 
               
               
                   
               
               
                 Sensory evaluation of chocolate flavored beverage 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 Nature of Participants: 
                 Trained panel 
               
               
                 Number of Sessions 
                  1 
               
               
                 Number of Participants: 
                 15 
               
               
                 Test Design: 
                 Balanced, randomized within set.  
               
               
                   
                 Blind 
               
               
                 Sensory Test Method: 
                 Intensity and acceptance ratings 
               
               
                 Environmental Condition 
                 Standard booth lighting 
               
               
                 Attributes and Scales: 
                   
               
            
           
           
               
            
               
                 Overall Acceptance on a 10-pt hedonic scale where 10 = Extremely Like  
               
               
                 and 0 = Extremely Dislike 
               
               
                 Overall Liking, sweet intensity, bitterness, cocoa flavor, dairy note, sweet  
               
               
                 aftertaste and bitter aftertaste. 10-pt continuous intensity scale where  
               
               
                 0 = Imperceptible and 10 = Extremely Pronounced 
               
            
           
           
               
               
            
               
                 Statistical Analysis: 
                 ANOVA (by Block) with  
               
               
                   
                 Post Hoc Duncan&#39;s Test 
               
               
                 Sample Size 
                 ~1.5 oz. in a clear capped plastic cup 
               
               
                 Serving Temperature 
                 Refrigerated temperature (~45° F.) 
               
               
                 Serving/Panelists 
                 Samples served simultaneously.  
               
               
                 Instruction: 
                 Panelists instructed to evaluate  
               
               
                   
                 each sample. 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 23 
               
             
            
               
                   
               
               
                 Sensory Results 
               
            
           
           
               
               
               
            
               
                   
                   
                 Test with  
               
               
                   
                 Control  
                 Reduced 
               
               
                   
                 with Full 
                 Sugar and  
               
               
                   
                 Amount of  
                 Cocoa with 
               
               
                   
                 Sugar and 
                 Stevia  
               
               
                 Attribute 
                 Cocoa 
                 Extract FMP 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Sweet intensity 
                 7.05 
                 7.26 
               
               
                 Bitterness 
                 0.65 
                 0.55 
               
               
                 Cocoa flavor 
                 4.4 
                 4.28 
               
               
                 Dairy note 
                 4.91 
                 5.13 
               
               
                 Sweet aftertaste 
                 1.24 
                 1.43 
               
               
                 Bitter aftertaste 
                 0.21 
                 0.16 
               
               
                 Overall liking 
                 7.43 
                 7.12 
               
               
                   
               
            
           
         
       
     
     As seen in Table 23, the test product with reduced sugar and cocoa, and containing  Stevia  extract FMP, in this case PSB-5005, had statistically similar overall liking and mean cocoa flavor intensity results as compared to a full-cocoa formulation. The test product containing  Stevia  extract FMP had lower bitterness attribute and bitter aftertaste ratings compared to the control product made without  Stevia  extract. The dairy note was rated higher in the test product compared to the control product. From these results it can be seen that a reduction in cocoa and sugar content in a dairy beverage can be suitably accomplished using a  Stevia  extract FMP, such as PSB-5005, and unexpectedly with a decrease in bitterness which is typically associated with  Stevia  ingredients. 
     Example 14: Effect of  Stevia  Extract FMP on Flavor Modification of Vanilla-Flavored Dairy Product 
     A 50% sugar-reduced vanilla yogurt was developed to determine the flavor modification effect of  Stevia  extract flavor with modifying properties (FMP) in a reduced-sugar vanilla-flavored dairy product. The two samples as shown in Table 24 included: 1) control sample with 180 ppm steviol glycoside sweetener, and 2) test sample with 180 ppm steviol glycoside sweetener and 100 ppm of  Stevia  extract FMP. 
     
       
         
           
               
             
               
                 TABLE 24 
               
             
            
               
                   
               
               
                 Sugar Reduced Vanilla-Flavored Dairy Product (Vanilla Yogurt) 
               
            
           
           
               
               
               
            
               
                   
                   
                 Test with  
               
               
                   
                 Control  
                 Reduced 
               
               
                   
                 180 ppm  
                 Sugar and  
               
               
                   
                 steviol 
                 Cocoa with 
               
               
                   
                 glycoside  
                 Stevia  
               
               
                   
                 sweetener 
                 Extract FMP 
               
               
                   
               
            
           
           
               
               
               
            
               
                 Plain nonfat yogurt 
                 96.132 
                 96.122 
               
               
                 Sugar 
                 3.750 
                 3.750 
               
               
                 Vanilla Flavor 
                 0.100 
                 0.100 
               
               
                 Steviol Glycoside 
                 0.018 
                 0.018 
               
               
                 Stevia Extract  
                 — 
                 0.010 
               
               
                 (PCS 5001) 
                   
                   
               
               
                 Total 
                 100.000 
                 100.000 
               
               
                   
               
            
           
         
       
     
     A 30 member panel evaluated samples of vanilla yogurt for overall acceptance and attribute intensities (sweet intensity, bitterness, vanilla flavor, dairy, astringency, sweet aftertaste and bitter aftertaste). Table 25 lists the sensory evaluation parameters. 
     
       
         
           
               
             
               
                 TABLE 25 
               
               
                   
               
               
                 Sensory evaluation of vanilla flavored dairy product 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 Nature of Participants: 
                 Trained sensory panel 
               
               
                 Number of Sessions 
                  1 
               
               
                 Number of Participants: 
                 30 
               
               
                 Test Design: 
                 Balanced, randomized within set.  
               
               
                   
                 Blind 
               
               
                 Sensory Test Method: 
                 Intensity and acceptance ratings 
               
               
                 Environmental Condition 
                 Standard booth lighting 
               
               
                 Attributes and Scales: 
                   
               
            
           
           
               
            
               
                 Overall Acceptance on a 10-pt hedonic scale where 10 = Extremely Like  
               
               
                 and 0 = Extremely Dislike 
               
               
                 Overall Liking, sweet intensity, bitterness, vanilla flavor, dairy, astringency, 
               
               
                 sweet aftertaste and bitter aftertaste. 10-pt continuous intensity scale where  
               
               
                 0 = Imperceptible and 10 = Extremely Pronounced 
               
            
           
           
               
               
            
               
                 Statistical Analysis: 
                 ANOVA (by Block) with  
               
               
                   
                 Post Hoc Duncan&#39;s Test 
               
               
                 Sample Size 
                 ~1.5 oz. in a clear capped plastic cup 
               
               
                 Serving Temperature 
                 Refrigerated temperature (~45° F.) 
               
               
                 Serving/Panelists 
                 Samples served simultaneously.  
               
               
                 Instruction: 
                 Panelists instructed to evaluate  
               
               
                   
                 each sample. 
               
               
                   
               
            
           
         
       
     
     Table 26 shows the sensory results for the control and test products. At 95% confidence, the test sample containing  Stevia  extract FMP was significantly higher for sweet intensity and vanilla flavor and significantly and unexpectedly lower in bitterness, astringency and sweet aftertaste. At 90% confidence, the test sample was higher in dairy and had higher overall liking.  FIG. 15  illustrates this comparison. 
     
       
         
           
               
             
               
                 TABLE 26 
               
             
            
               
                   
               
               
                 Summary of the overall acceptance and mean attribute 
               
               
                 intensity results for vanilla-flavored dairy product 
               
               
                 (vanilla yogurt) with stevia extract 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Test with  
                   
                   
               
               
                   
                   
                 180 ppm 
                   
                   
               
               
                   
                   
                 steviol  
                   
                   
               
               
                   
                 Control  
                 glycoside 
                   
                   
               
               
                   
                 180 ppm 
                 sweetener and  
                   
                   
               
               
                   
                 steviol  
                 100 ppm  
                   
                   
               
               
                   
                 glycoside 
                 stevia extract 
                   
                   
               
               
                 Attribute 
                 sweetener 
                 FMP 
                 p-value 
                 Sig 
               
               
                   
               
               
                 Sweet intensity 
                 6.88 
                 7.05 
                 0.0281 
                 *** 
               
               
                 Bitterness 
                 2.53 
                 1.74 
                 0.0374 
                 *** 
               
               
                 Vanilla flavor 
                 4.95 
                 5.87 
                 0.0452 
                 *** 
               
               
                 Dairy 
                 4.78 
                 5.65 
                 0.0507 
                 ** 
               
               
                 Astringency 
                 2.36 
                 1.63 
                 0.0407 
                 *** 
               
               
                 Sweet aftertaste 
                 1.73 
                 0.95 
                 0.0144 
                 *** 
               
               
                 Bitter aftertaste 
                 1.55 
                 0.94 
                 0.0682 
                 ** 
               
               
                 Overall liking 
                 5.26 
                 6.37 
                 0.0629 
                 ** 
               
               
                   
               
            
           
         
       
     
     Example 15: Usage Levels of  Stevia  Extract FMP 
     Useful or maximum usage levels of  Stevia  extract FMPs were evaluated. To be useful as a flavor with modifying properties, the level of use of the  Stevia  extract must be below a certain sweetness detection threshold in a particular food or beverage product. To determine this threshold, a sensory evaluation is conducted with a full sugar product as the control, and a test product containing different levels of the  Stevia  extract FMP. Sensory panel members are then asked to identify which product is sweeter. 
     Using the Flavor and Extract Manufacturers&#39; Association (FEMA) guidance document called “Guidance for the Sensory Testing of Flavorings with Modifying Properties within the FEMA GRAS™ Program, 2013”, the recognition threshold was determined using a 2-alternatve forced choice (2-AFC) methodology, as described in Table 27. 
     
       
         
           
               
             
               
                 TABLE 27 
               
               
                   
               
               
                 Sensory evaluation to determine usage levels 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
            
               
                 Nature of Participants: 
                 Company employees 
               
               
                 Number of Sessions 
                  1 
               
               
                 Number of Participants: 
                 30 
               
               
                 Test Design: 
                 2- AFC, Balanced, randomized  
               
               
                   
                 within pair. Blind 
               
               
                 Sensory Test Method: 
                 Intensity ratings 
               
               
                 Environmental Condition 
                 Standard booth lighting 
               
               
                 Attributes and Scales: 
                 Which sample is sweeter? 
               
               
                 Statistical Analysis: 
                 Paired comparison Test 
               
               
                 Sample Size 
                 ~1.5 oz. in a clear capped plastic cup 
               
               
                 Serving Temperature 
                 Refrigerated temperature (~42° F.)  
               
               
                   
                 or room temperature, depending  
               
               
                   
                 on sample requirements 
               
               
                 Serving/Panelists 
                 Samples served simultaneously.  
               
               
                 Instruction: 
                 Panelists instructed to read ingredient  
               
               
                   
                 statement, evaluate each sample. 
               
               
                   
               
            
           
         
       
     
     Usage levels for  Stevia  extract FMP are determined by those levels at which the  Stevia  extract FMP provides a sweetness perception that is significantly less than the full sugar control. For products other than baked goods and breakfast cereals, the sugar level in the control product was 1.5%. In baked goods, the sugar level in the control product was 4%, and in breakfast cereals, the sugar level in the control product was 3%. The test products contained no added sugar and contain various levels of  Stevia  extract FMP. 
     Table 28 shows usage levels of  Stevia  extract FMP in various food and beverage applications as determined using the FEMA sensory testing guidance. 
     
       
         
           
               
             
               
                 TABLE 28 
               
             
            
               
                   
               
               
                 Usage levels of stevia extract FMP 
               
            
           
           
               
               
            
               
                 Category 
                 Usage Level (ppm) 
               
               
                   
               
               
                 Baked Goods 
                 500 
               
               
                 Beverages, Non-Alcoholic 
                 110 
               
               
                 Beverages, Alcoholic 
                 130 
               
               
                 Breakfast Cereals 
                 600 
               
               
                 Chewing Gum 
                 100 
               
               
                 Condiments and Relishes 
                 100 
               
               
                 Confections and Frostings 
                 100 
               
               
                 Fats and Oils 
                 180 
               
               
                 Frozen Dairy 
                 100 
               
               
                 Fruit Ices 
                 100 
               
               
                 Gelatins and Puddings 
                 100 
               
               
                 Gravies 
                 100 
               
               
                 Hard Candy 
                 100 
               
               
                 Imitation Dairy 
                 165 
               
               
                 Instant Coffee and Tea 
                 200 
               
               
                 Jams and Jellies 
                 100 
               
               
                 Milk Products 
                 165 
               
               
                 Nut Products 
                 230 
               
               
                 Processed Fruits 
                 100 
               
               
                 Processed Vegetables 
                 100 
               
               
                 Seasonings and Flavors 
                 230 
               
               
                 Snack Foods 
                 230 
               
               
                 Soft Candy 
                 100 
               
               
                 Soups 
                 100 
               
               
                 Sugar Substitutes 
                 100 
               
               
                 Sweet Sauces 
                 100 
               
               
                   
               
            
           
         
       
     
     It was unexpectedly discovered that  Stevia  extract FMPs can be used at various levels to favorably impact the taste and flavor profile of a food or beverage product while having little or no detectable sweetness perception in that product. These usage levels serve as examples of use, and other usage levels of the  Stevia  extract FMP in various consumable products are contemplated by this invention. 
     Although various embodiments of the present invention have been disclosed here for purposes of illustration, it should be understood that a variety of changes, modifications and substitutions may be incorporated without departing from either the spirit or the scope of the invention.