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

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.

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.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0029] FIG. 1 shows the chemical structure of the diterpene glycosides of Stevia rebaudiana.

[0030] 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.

[0031] 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.

[0032] 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.

[0033] FIG. 5 is a bar graph showing the effect of stevia extract on the flavor profile of roasted peanuts.

[0034] FIG. 6 is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of stevia extract to tomato ketchup.

[0035] FIG. 7 is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of stevia extract to chocolate milk.

[0036] 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.

[0037] FIG. 9 is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of stevia extract to tortilla chips.

[0038] FIG. 10 is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of stevia extract to beef jerky.

[0039] FIG. 11 is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of stevia extract to brown gravy.

[0040] FIG. 12 is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of stevia extract to chocolate milk.

[0041] FIG. 13 is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of stevia extract to vanilla custard.

[0042] FIG. 14 is a bar graph showing the modification of flavor and sweetness profiles caused by the addition of stevia extract to chocolate milk.

[0043] 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

[0044] Embodiments of the present invention are described in the following examples.

EXAMPLES

Example 1A: Detection of Concentration Threshold for Sweetness Recognition

[0045] 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.

[0046] 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.

[0047] 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-US-00005 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

[0048] 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.

[0049] 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-US-00006 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

[0050] 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-US-00007 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-US-00008 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.

[0051] 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

[0052] 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-US-00009 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

[0053] 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

[0054] 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.

[0055] 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-US-00010 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-US-00011 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

[0056] 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-US-00012 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

[0057] 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

[0058] 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-US-00013 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-US-00014 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.

[0059] 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

[0060] 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-US-00015 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

[0061] 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

[0062] 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).

[0063] 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-US-00016 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

[0064] 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).

[0065] 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-US-00017 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

[0066] 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).

[0067] 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

[0068] 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-US-00018 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

[0069] 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-US-00019 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 .sup. 6.82 b .sup. 7.70 a 0.0482 *** Dairy Note .sup. 3.61 b .sup. 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.sup.  0.35.sup.  0.2473 NS Bitter Aftertaste 0.71 a 0.55 b 0.1824 * Sweet Aftertaste 2.66.sup.  2.82.sup.  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)

[0070] 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.

[0071] 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-US-00020 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

[0072] 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-US-00021 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.sup.  0.5   0.3001 NS Bitter Aftertaste 0.36.sup.  0.38.sup.  0.7692 NS Sweet Aftertaste 2.23.sup.  2.24.sup.  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

[0073] 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-US-00022 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

[0074] 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.

[0075] 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-US-00023 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-US-00024 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

[0076] 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

[0077] 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-US-00025 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

[0078] 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-US-00026 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.

[0079] 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-US-00027 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

[0080] 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.

[0081] 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-US-00028 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.

[0082] 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.

[0083] Table 28 shows usage levels of stevia extract FMP in various food and beverage applications as determined using the FEMA sensory testing guidance.

TABLE-US-00029 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

[0084] 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.

[0085] 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.