STEVIA EXTRACT CONTAINING SELECTED STEVIOL GLYCOSIDES AS FLAVOR, SALTY AND SWEETNESS PROFILE MODIFIER

Information

  • Patent Application
  • 20170339994
  • Publication Number
    20170339994
  • Date Filed
    June 09, 2017
    7 years ago
  • Date Published
    November 30, 2017
    6 years ago
Abstract
Stevia extracts with selected major steviol glycosides (Reb A, stevioside, Reb D, Reb C) and minor steviol glycosides and glycosylated diterpene derivative plant molecules, derived from Stevia rebaudiana plant are found to improve the perception of flavor and taste perception, which includes the sweet, savory and salty perception in a wide range of food and beverage applications.
Description
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]-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.


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







TMelt,
Mol.
Solubility in
Relative
Quality


Name
Formula
° C.
Weight
water, %
sweetness
of taste





















Steviol
C20H30O3
212-213
318.45
ND
ND
Very bitter


Steviolmonoside
C26H40O8
ND
480.58
ND
ND
ND


Stevioside
C38H60O18
196-198
804.88
0.13
210
Bitter


Rebaudioside A
C44H70O23
242-244
967.01
0.80
200-400
Less Bitter


Rebaudioside B
C38H60O18
193-195
804.88
0.10
150
Bitter


Rebaudioside C
C44H70O22
215-217
951.01
0.21
30
Bitter


Rebaudioside D
C50H80O28
248-249
1129.15
1.00
220
Like sucrose


Rebaudioside E
C44H70O23
205-207
967.01
1.70
170
Like sucrose


Rebaudioside F
C43H68O22
ND
936.99
ND
ND
ND


Dulcoside A
C38H60O17
193-195
788.87
0.58
30
Very bitter


Steviolbioside
C32H50O13
188-192
642.73
0.03
90
Unpleasant


Rubusoside
C32H50O13
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)














Common
Trivial
Mol.





#
name
formula
Wt.
R1
R2
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)
Kohda et







Glcβ1-
al. (1976)


1.5
Stevioside
SvG3
805
Glcβ1-
Glcβ(1-2)
Bridel &







Glcβ1-
Lavielle








(1931)


1.6
Stevioside A
SvG3
805
Glcβ(1-2)
Glcβ1-
Wu et al.






Glcβ1-

(2012)


1.7
Rebaudioside B
SvG3
805
H
Glcβ(1-2)
Kohda et







[Glcβ(1-3)]
al. (1976)







Glcβ1-



1.8
Rebaudioside G
SvG3
805
Glcβ1-
Glcβ(1-3)
Ohta et al.







Glcβ1-
(2010)


1.9
Stevioside B
SvG3
805
Glcβ(1-3)
Glcβ1-
Chaturvedula






Glcβ1-

& Zamora








(2014)


1.10
Rebaudioside E
SvG4
967
Glcβ(1-2)
Glcβ(1-2)
Sakamoto






Glcβ1-
Glcβ1-
et al.








(1977a)


1.11
Rebaudioside A
SvG4
967
Glcβ1-
Glcβ(1-2)
Kohda et







[Glcβ(1-3)]
al. (1976)







Glcβ1-



1.12
Rebaudioside A2
SvG4
967
Glcβ1-
Glcβ(1-6)
Chaturvedula







Glcβ(1-2)
& Prakash







Glcβ1-
(2011d)


1.13
Rebaudioside D
SvG5
1129
Glcβ(1-2)
Glcβ(1-2)
Sakamoto






Glcβ1-
[Glcβ(1-3)]
et al.







Glcβ1-
(1977a)


1.14
Rebaudioside I
SvG5
1129
Glcβ(1-3)
Glcβ(1-2)
Ohta et al.






Glcβ1-
[Glcβ(1-3)]
(2010)







Glcβ1-



1.15
Rebaudioside L
SvG5
1129
Glcβ1-
Glcβ(1-6)
Ohta et al.







Glcβ(1-2)
(2010)







[Glcβ(1-3)]








Glcβ1-



1.16
Rebaudioside Q2
SvG5
1129
Glcα(1-2)
Glcβ(1-2)
Chaturvedula






Glcα(1-4)
Glcβ1-
& Prakash






Glcβ1-

(2011c)


1.17
Rebaudioside Q
SvG5
1129
Glcβ1-
Glcα(1-4)








Glcβ(1-2)








[Glcβ(1-3)]








Glcβ1-



1.18
Rebaudioside I2
SvG5
1129
Glcβ1-
Glcα(1-3)
Chaturvedula







Glcβ(1-2)
et al.







[Glcβ(1-3)]
(2011c)







Glcβ1-



1.19
Rebaudioside Q3
SvG5
1129
Glcβ1-
Glcα(1-4)
Chaturvedula







Glcβ(1-3)
et al.







[Glcβ(1-2)]
(2011c)







Glcβ1-



1.20
Rebaudioside I3
SvG5
1129
Glcβ(1-2)
Glcβ(1-2)
Chaturvedula






[Glcβ(1-6)]
Glcβ1-
et al.






Glcβ1-

(2011c)


1.21
Rebaudioside M
SvG6
1291
Glcβ(1-2)
Glcβ(1-2)
Ohta et al.






[Glcβ(1-3)]
[Glcβ(1-3)]
(2010)






Glcβ1-
Glcβ1-








2. Steviol + Rhamnose + Glucose (SvR1Gn)













2.1
Dulcoside A
SvR1G2
789
Glcβ1-
Rhaα(1-2)
Kobayashi







Glcβ1-
et al.








(1977)


2.2
Dulcoside B
SvR1G2
789
H
Rhaα(1-2)
Ohta et al.







[Glcβ(1-3)]
(2010)







Glcβ1-



2.3
Rebaudioside C
SvR1G3
951
Glcβ1-
Rhaα(1-2)
Sakamoto







[Glcβ(1-3)]
et al.







Glcβ1-
(1977b)


2.4
Rebaudioside
SvR1G3
951
Rhaα(1-2)
Glcβ(1-2)
Purkayastha



C2a


Glcβ1-
Glcβ1-
(2016)


2.5
Rebaudioside S
SvR1G3
951
Rhaα(1-2)
Glcα(1-2)
Ibrahim et






Glcβ1-
Glcβ1-
al (2016)


2.6
Rebaudioside H
SvR1G4
1112
Glcβ1-
Glcβ(1-3)
Ohta et al.







Rhaα(1-2)]
(2010)







[Glcβ(1-3)]








Glcβ1-



2.7
Rebaudioside K
SvR1G4
1112
Glcβ(1-2)
Rhaα(1-2)
Ohta et al.






Glcβ1-
[Glcβ(1-3)]
(2010)







Glcβ1-



2.8
Rebaudioside J
SvR1G4
1112
Rhaα(1-2)
Glcβ(1-2)
Ohta et al.






Glcβ1-
[Glcβ(1-3)]
(2010)







Glcβ1-



2.9
Rebaudioside N
SvR1G5
1274
Rhaα(1-2)
Glcβ(1-2)
Ohta et al.






[Glcβ(1-3)]
[Glcβ(1-3)]
(2010)






Glcβ1-
Glcβ1-



2.10
Rebaudioside O
SvR1G6
1436
Glcβ(1-3)
Glcβ(1-2)
Ohta et al.






Rhaα(1-2)
[Glcβ(1-3)]
(2010)






[Glcβ(1-3)]
Glcβ1-







Glcβ1-




2.11
Rebaudioside
SvR1G6
1436
Glcβ(1-4*)
Glcβ(1-2)
Purkayastha



O2a


Rhaα(1-2)
[Glcβ(1-3)]
(2016)






[Glcβ(1-3)]
Glcβ1-







Glcβ1-




2.12
Rebaudioside
SvR1G4
1112
Glcβ(1-6)
Rhaα(1-2)
Purkayastha



K2a


Glcβ1-
[Glcβ(1-3)]
(2016)







Glcβ1-








3. Steviol + Xylose + Glucose (SvX1Gn)













3.1
Stevioside F
SvX1G2
775
Glcβ1-
Xylβ(1-2)
Chaturvedula







Glcβ1-
& Prakash








(2011b)


3.2
Rebaudioside F
SvX1G3
937
Glcβ1-
Xylβ(1-2)
Starratt et







[Glcβ(1-3)]
al. (2002)







Glcβ1-



3.3
Rebaudioside F2
SvX1G3
937
Glcβ1-
Glcβ(1-2)
Chaturvedula







[Xylβ(1-3)]
& Prakash







Glcβ1-
(2011b)


3.4
Rebaudioside F3
SvX1G3
937
Xylβ(1-6)
Glcβ(1-2)
Chaturvedula






Glcβ1-
Glcβ1-
et al.








(2011d)


3.5
Rebaudioside R
SvX1G3
937
Glcβ1-
Glcβ(1-2)
Ibrahim et







[Glcβ(1-3)]
al (2016)







Xylβ1-



3.6
Rebaudioside
SvX1G4
1099
Xylβ(1-2)
Glcβ(1-2)
Purkayastha



Ua


Glcβ1-
[Glcβ(1-3)]
(2016)







Glcβ1-



3.7
Rebaudioside
SvX1G4
1099
Xylβ(1-2*)
Glcβ(1-2)
Purkayastha



U2a


[Glcβ(1-3)]
Glcβ1-
(2016)






Glcβ1-




3.8
Rebaudioside
SvX1G5
1261
Glcβ(1-2)
Xylβ(1-2*)
Purkayastha



Va


[Glcβ(1-3)]
[Glcβ(1-3)]
(2016)






Glcβ1-
Glcβ1-



3.9
Rebaudioside
SvX1G5
1261
Xylβ(1-2)
Glcβ(1-2)
Prakash &



V2a


[Glcβ(1-3)]
[Glcβ(1-3)]
Chaturvedula






Glcβ1-
Glcβ1-
(2013)







4. Steviol + Arabinose + Glucose (SvA1Gn)













4.1
Rebaudioside
SvA1G4
1098
Glcβ(1-2)
Glcβ(1-2)
Purkayastha



Wa


[Araβ(1-3*)]
Glcβ1-
(2016)






Glcβ1-




4.2
Rebaudioside
SvA1G4
1098
Araβ(1-2*)
Glcβ(1-2)
Purkayastha



W2a


Glcβ1-
[Glcβ(1-3)]
(2016)







Glcβ1-



4.3
Rebaudioside
SvA1G4
1098
Araβ(1-6)
Glcβ(1-2)
Purkayastha



W3a


Glcβ1-
[Glcβ(1-3)]
(2016)







Glcβ1-



4.4
Rebaudioside
SvA1G5
1260
Glcβ(1-2)
Glcβ(1-2)
Purkayastha



Ya


[Araβ(1-3*)]
[Glcβ(1-3)]
(2016)






Glcβ1-
Glcβ1-








5. Steviol + Fructose + Glucose (SvF1Gn)













5.1
Rebaudioside A3
SvF1G3
967
Glcβ1-
Glcβ(1-2)
Chaturvedula







[Fruβ(1-3)]
et al.







Glcβ1-
(2011b)







6. Steviol + galactose + Glucose (SvGa1Gn)













6.1
Rebaudioside
SvGa1G4
1128
Glcβ(1-2*)
Glcβ(1-2)
Purkayastha



Ta


Glcβ1
[Glcβ(1-3)]
(2016){circumflex over ( )}







Glcβ1-








7. Steviol + de-oxy glucose + Glucose (SvdG1Gn)













7.1
Stevioside D
SvdG1G2
789
Glcβ1-
6-deoxyGlcβ
Chaturvedula







(1-2)
& Prakash







Glcβ1-
(2011a)


7.2
Stevisoide E
SvdG1G3
951
Glcβ1-
6-deoxyGlcβ
Chaturvedula







(1-2)
& Prakash







[Glcβ(1-3)]
(2011a)







Glcβ1-



7.3
Stevioside E2
SvdG1G3
951
6-deoxyGlcβ1-
Glcβ(1-2)
Chaturvedula







[Glcβ(1-3)]
et al.







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
Stigmasterol, β-sitosterol, Campesterol


derivatives


Flavonoids
Glucosyl-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













and



Steviol Glycosides*, %

related





















Stevia

Stevio-



Dulco-
Rubu-

Steviol-



TSG*
plant


Extracts
Reb A
side
Reb D
Reb F
Reb C
side A
soside
Reb B
bioside
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, 199-209 (2010)).


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]-L-phenylalanine 1-methyl ester, N—[N-[3-(3-hydroxy-4-methoxyphenyl)-3-methylbutyl]-L-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.












Sugar

Stevia





Comparison of sweetness
solution
Extract


perception of STEVIA
(1.5%)
solution

Signif-


EXTRACT in water
sweeter?
sweeter?
P-Value
icance














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)












CSD
CSD





sample
sample


Sweetness perception of
with Sugar
with Stevia

Signif-



STEVIA EXTACT in CSD

sweeter?
Sweeter?
P-Value
icance














PCS-5001: 110 ppm,
23
7
0.0052
***


N = 30


PCS-5001: 120 ppm,
20
16
0.677
NS


N = 36


PCS-1015: 70 ppm,
21
9
0.0428
***


N = 30


PCS-1015: 90 ppm,
12
18
0.3616
NS


N = 30









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% Sugar
Reduction with
Reduction with


FORMULA
Reduction
PCS-5001
PCS-1015













Water
91.68
91.67
91.67


Sugar
7.89
7.89
7.89


Cola Flavor -
0.375
0.375
0.375


Flavor Systems


Phosphoric Acid
0.0333
0.0333
0.0333


85%


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'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



Instruction:
instructed to 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











Reduced
Reduce Sugar Tea
Reduce Sugar Tea



Sugar Tea
with PCS-5001
with PCS-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


0.0120


PCS-5001



Stevia Extract



0.0080


PCS-1015


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



Steviol
Glycoside +
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's Test










Sample Size
~1.5 oz. in a clear capped plastic









cup










Serving Temperature
Room temperature (~70° F.)



Serving/Panelists
Samples served simultaneously.



Instruction:
Panelists evaluate each 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 with
Reduction with stevia


Dairy Formula
steviol glycoside
extract and stevia glycoside












2% Reduced fat
96.5803
96.5753


Milk


Sugar
2.40
2.40


Cocoa Powder
0.80
0.80


Palsgaard 150
0.20
0.20


ChoMilk


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
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, 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'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



Instruction:
instructed to read 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 & bitter aftertaste).









TABLE 14







Effect of PCS-5001(stevia extract) on baked goods











Steviol glycoside



Steviol Glycoside
w/120 ppm


Ingredients
(400 ppm) 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 & 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 & 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%
50%
50%



Total Sugar
Total Sugar
Total Sugar



Reduction with
Reduction with
Reduction with



PureCircle
PC Alpha &
PC Alpha &


Dairy Formula
Alpha
PCS-1015
PCS-1015













2% Reduced fat Milk
96.5803
96.5743
96.5753


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 Alpha
0.0197
0.0197
0.0197


PCS-1015 (MLD-1)

0.0070
0.0080


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 of
70 ppm of





Alpha Only
MLD-1 w/


Attribute
(Control)
PC 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 of
80 ppm of





Alpha Only
MLD-1 w/


Attribute
(Control)
PC 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











Test with



Control with
PureCircle



PureCircle
Alpha w/



Alpha

stevia extract














Milk (1% fat)
94.27
94.27


Sucrose
4.00
4.00


Starch Perma Flo Tate & Lyle
1.25
1.25


TIC Carrageenan
0.09
0.09


Salt
0.06
0.06


ROHA Beta Carotene
0.05
0.05


French Vanilla Flavor UV 420-066-7
0.15
0.15


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 of





Alpha Only
MLD-1 with


Attribute
(Control)
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










Control with Full
Test with Reduced



Amount of Sugar
Sugar and Cocoa



and Cocoa
with Stevia Extract













Milk, 2% milkfat
85.200
86.554


Sugar
8.00
6.80


Hot Water, 190 F.
6
6


Natural 10/12 Cocoa Powder
0.80
0.64



Stevia Extract FMP (PSB-5005)


0.006


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
Trained panel


Participants:


Number of Sessions
 1


Number of
15


Participants:


Test Design:
Balanced, randomized within set.



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, 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'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


Instruction:
instructed to evaluate each sample.
















TABLE 23







Sensory Results










Control with Full
Test with Reduced



Amount of Sugar and
Sugar and Cocoa with


Attribute
Cocoa

Stevia 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)










Control 180 ppm
Test with Reduced



steviol glycoside
Sugar and Cocoa with



sweetener

Stevia 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 (PCS 5001)


0.010


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
Trained sensory panel


Participants:


Number of Sessions
 1


Number of
30


Participants:


Test Design:
Balanced, randomized within set. 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, 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'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


Instruction:
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 glycoside



Control 180 ppm
sweetener and 100



steviol glycoside
ppm 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' 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-alternative 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. Panelists


Instruction:
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.

Claims
  • 1. A stevia extract flavor with modifying properties (FMP) comprising: (a) rebaudioside A at a level of up to about 30% by weight;(b) stevioside at a level of up to about 12% by weight; and(c) minor steviol glycosides and plant molecules at a level of up to 55% by weight,wherein the stevia extract FMP modifies the flavor and sweetness profile of a consumable product without imparting significant sweetness to the consumable product.
  • 2. The stevia extract FMP of claim 1, further comprising up to about 25% rebaudioside C by weight and up to about 20% rebaudioside D by weight.
  • 3. The stevia extract FMP of claim 1, further comprising up to about 6% rebaudioside E by weight, up to about 8% rebaudioside N by weight, and up to about 8% rebaudioside O by weight.
  • 4. The stevia extract FMP of claim 1, wherein the minor steviol glycosides comprise one or more selected from the group consisting of: rebaudioside G; rebaudioside H; rebaudioside I, rebaudioside J; rebaudioside K; and rebaudioside L.
  • 5. The stevia extract FMP of claim 1, wherein the plant glycosides comprise one or more selected from the group consisting of: glucosyl-4′-O-apigenin; glucosyl-7-O-luteolin; rhamnosyl-3-O-kaempferol; glucosyl-3-O-quercitin; and arabinosyl-3-O-quercetin.
  • 6. A method for modifying a flavor of a consumable product, comprising adding the stevia extract FMP of claim 1.
  • 7. The method of claim 6, wherein the stevia extract FMP is added to the consumable product in an amount below a stevia extract FMP sweetness detection threshold of about 110 ppm.
  • 8. The method of claim 6, wherein the stevia extract FMP is added to the consumable product in an amount of up to 100 ppm.
  • 9. The method of claim 6, wherein the stevia extract FMP is added to the consumable product in an amount ranging from about 60 ppm to 100 ppm.
  • 10. The method of claim 6, wherein the flavor of the consumable product is modified by reducing a bitterness of the consumable product as compared to a control product that does not contain the stevia extract FMP.
  • 11. The method of claim 6, wherein the flavor of the consumable product is modified by reducing a bitter aftertaste of the consumable product as compared to a control product that does not contain the stevia extract FMP.
RELATED APPLICATIONS

This application 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.

Provisional Applications (2)
Number Date Country
61832451 Jun 2013 US
61942331 Feb 2014 US
Continuation in Parts (1)
Number Date Country
Parent 14896022 Dec 2015 US
Child 15619107 US