INGREDIENT SYSTEMS COMPRISING TREHALOSE, FOOD PRODUCTS CONTAINING TREHALOSE, AND METHODS OF MAKING SAME

RELATED APPLICATIONS This application claims priority under 35 U.S.C. §119(e) of U.S. Application No. 60/602,878, filed August 20, 2004. FIELD OF INVENTION The present invention relates to salt-substitute compositions comprising trehalose, food products comprising such compositions and methods of preparing such food products. The salt-substitute compositions include ingredient systems comprising trehalose, potassium chloride, and/or sodium chloride, and the food products comprising the ingredient system include less than 1.5% trehalose by weight. The present invention also relates to ingredient systems comprising trehalose, which systems can be useful for reducing the level of sodium in food products; food products comprising less than 1.5% by weight of trehalose, which food products can have a reduced level of sodium; and methods of reducing the amount of sodium in food products. As used herein, "trehalose" refers to alpha-D-glucopyranosyl alpha-D- glucopyranoside in a form that is suitable for human consumption. BACKGROUND Salt substitute compositions containing potassium chloride with and without sodium chloride have been proposed for people who seek to reduce the amount of sodium chloride in their diet due to medical necessity or for other reasons. However, potassium chloride can be associated with a bitter, metallic taste and salt-substitute compositions comprising potassium chloride can therefore lack palatability and/or lack satisfactory similarity to common salt (i.e. sodium chloride) in taste. U.S. Patent No. 6,159,529 ("the '529 patent") relates to a method for enhancing the salty-taste and/or delicious-taste of food products containing sodium chloride by incorporating trehalose into the food product in the range of 1.5% to 12% on a dry solid basis. The '529 patent states that trehalose content below 1.5% does not sufficiently enhance the level of salty- and delicious-taste. CGL03/0470WO

SUMMARY The present invention relates to methods for reducing the sodium content of finished food products by adding trehalose to suitable food products in an amount ranging from greater than 0% to less than 1.5% by weight of the finished food product. The method can further include adding potassium or sodium or combinations thereof to the suitable food products. In some embodiments, the finished food product has less sodium than a control product, yet the finished food product has a similar salty taste and flavor as the control product. The present invention also relates to ingredient systems for achieving salt (i.e. sodium) reduction in suitable food products. The ingredient systems comprise trehalose and can include sodium, potassium, or combinations thereof. For purposes of this specification, "suitable food products" specifically refer to liquid systems, including food systems that have a liquid component including salt, food systems having topically applied salt, and cheese spreads. Examples of suitable finished food systems include marinated meats such as marinated whole muscle meat (e.g. beef, pork, poultry); Soups, Sauces, Stews; Bullion, Pasta and Sauce; Prepared Instant Pasta; Prepared Instant Rice; Prepared Meat seasoning Mixes; Gravy; Marinades; Glazes; Vegetable Side dishes; Potato Side Dishes; Dressings; Salsa; Meat and Vegetable Stocks; Soy Sauce (wherein the culture is not capable of metabolizing trehalose); Meat and Vegetables Base; Potato Chips; Crackers; Pretzels; Tortilla Chips; French Fries; Popcorn; Extruded Snacks; Rice Cakes; Baked Goods with topically applied salt; topically applied seasonings, including: Seasoned Snacks (Potato chips, Corn Chips, Tortilla chips, Rice Cakes, Cheese Puffs); Seasoned Potatoes; Potato Sticks; Soy Crisps; Meat Rubs; cheese spreads including: process cheese base products (e.g. spreads, sauces; slices); dips (e.g. bean, dairy); cream cheese; cottage cheese; vegetables including: pickles; canned vegetables; canned tomato products (e.g. whole, paste, sauce); salsa; plain and prepared beans (e.g. legumes); sauerkraut, and, beverages including: vegetable juices and cocktails, and isotonic beverages. The salt reduction method may be used with any available application of the food products such as fresh, shelf stable, frozen and refrigerated applications. CGL03/0470WO

A "control food product" is a food product has substantially similar ingredients to a finished food product, except that the control food product includes a greater amount of sodium than the finished food product, and the control product does not include trehalose DESCRIPTION In one embodiment, the method of making a suitable finished food product comprises: (i) adding a total amount of sodium and potassium to a food product, wherein the total amount equals an amount of sodium added to a control food product; and, (ii) adding an amount of trehalose ranging from greater than 0% to less than 1.5% by weight of the finished food product to the food product. In one embodiment, the method of making a suitable finished food product comprises: (i) adding a total amount of sodium and potassium to a food product, wherein the total amount equals an amount of sodium added to a control food product, and further wherein the total amount of the sodium and potassium comprises from about 20% to about 50% potassium by weight; and, (ii) adding an amount of trehalose ranging from greater than 0% to less than 1.5% by weight of the finished food product to the food product, wherein the finished food product and the control product have a comparable salty taste. In one embodiment, the method of making a suitable finished food product comprises (i) adding a total amount of sodium and potassium to a food product, wherein the total amount equals an amount of sodium added to a control food product, and further wherein the total amount of the sodium and potassium comprises from about 20% to about 50% potassium by weight; and, (ii) adding an amount of trehalose ranging from greater than 0% to less than 1.5% by weight of the finished food product to the food product, wherein the finished food product and the control product have both a comparable salty taste and flavor. In one embodiment, the method comprises: adding a total amount of sodium, potassium, and trehalose to a suitable food product, wherein the total amount equals an amount of sodium added to a control food product, and wherein the amount of trehalose added is less than 1.5% by weight of the food product to the food product. In one embodiment, the method comprises making a finished food product, wherein making the finished food product comprises adding a total amount of sodium and trehalose to a suitable food product, wherein the finished food product has the same salty taste as a control CGL03/0470WO

product and further wherein the finished food product contains less than 1.5% trehalose by weight and less total sodium than the control product. In one embodiment, the method comprises making a finished food product, wherein making the finished food product comprises adding a total amount of potassium and trehalose to a suitable food product, wherein the finished food product has the same salty taste as a control product and further wherein the finished food product contains less than 1.5% trehalose by weight and less total sodium than the control product. In one embodiment, the method comprises making a finished food product, wherein making the finished food product comprises adding a total amount of sodium, potassium and trehalose to a suitable food product, wherein the finished food product has the same salty taste as a control product and further wherein the finished food product contains less than 1.5% trehalose by weight and less total sodium than the control product. The present invention also relates to ingredient systems useful for salt-substitute compositions. In one embodiment, the ingredient systems comprise sodium, potassium, and trehalose, wherein the ingredient systems when added to a suitable food product produces a finished food product having less than 1.5% trehalose by weight. In one embodiment, the ingredient systems comprise sodium chloride, potassium chloride, and trehalose, wherein the ingredient systems, when added to a suitable food product, deliver less than 1.5% trehalose by weight. In one embodiment, the ingredient systems comprise sodium, potassium, and trehalose, wherein the ingredient systems when added to a suitable food product produces a finished food product including less than 1.5% trehalose by weight, and wherein the finished food product contains an amount of sodium and potassium equal to the amount of sodium in a control food product. In one embodiment, the ingredient systems comprise sodium, potassium, and trehalose, wherein the ingredient systems when added to a suitable food product produces a finished food product including less than 1.5% trehalose by weight, and wherein the finished food product contains from about 20% to about 50% less sodium than a control food product, and further wherein the finished food product and the control food product have a comparable salty taste and comparable flavor. In some embodiments according to the present invention, the salt-substitute compositions comprise trehalose and sodium (for example sodium chloride). In some embodiments according to the present invention, the compositions comprise trehalose and CGL03/0470WO

potassium (for example potassium chloride). In some embodiments according to the present invention, the compositions comprise trehalose, sodium (for example sodium chloride) and potassium (for example potassium chloride). In some embodiments according to the present invention, the salt substitute compositions are used as seasonings for finished food products, and/or incorporated into finished food products. In some embodiments where the salt- substitute compositions are used as seasonings and/or incorporated into finished food products, the amount of trehalose in the finished food product is less than 1.5% by weight of the finished food product. In such embodiments, the amount of trehalose in the salt-substitute composition may, however, be far greater than 1.5% by weight of the salt-substitute composition. For example, the amount of trehalose in the salt -substitute composition may be determined by the following formula: 1.49 = S/(100-T)*T, explained in more detail below. In some embodiments, the amount of trehalose in the finished food product is less than 1.5% by weight of the finished food product and is such that the sweet taste of trehalose is imperceptible. It should be noted that the term "blend", when used to describe combinations of trehalose and other salts, is used interchangeably with the phrase "salt substitute composition comprising trehalose." Specifically, the term "blend" is not intended to mean, nor does it mean that the composition is or has been mixed, or that the composition is homogenous. According to one embodiment of the present invention, a composition comprises: (i) sodium chloride; and (ii) trehalose at an amount of less than 1.5% by weight of the composition. In some embodiments, the amount of trehalose may be up to about 1.2%, up to about 1.0%, up to about 0.75%, up to about 0.5%, up to 0.10% , up to about 0.075%; up to about 0.05%, up to about 0.0225%, up to about 0.015%; up to about 0.01%, up to about 0.008%, up to about 0.005%, up to about 0.003%, or up to 0.001% by weight of the composition. In other embodiments, the amount of trehalose may be about 0.001% to about 1.4%; about 0.003% to about 0.5%; about 0.003% to about 0.1%,; about 0.003% to about 0.075%; about 0.003% to about 0.05%; about 0.003% to about 0.0225%; about 0.003% to about 0.015%; about 0.003% to about 0.008%; about 0.015% to about 0.1%; about 0.015% to about 0.075%; about 0.015% to about 0.05%; or about 0.015% to about 0.0225% by weight of the composition. CGL03/0470WO

Another embodiment according to the present invention relates to a food product comprising sodium chloride and trehalose, wherein the amount of trehalose is less than 1.5% by weight of the food product. In some embodiments, the amount of trehalose in the blend is up to 99%, up to about 95%, up to about 90%, up to about 85%, up to about 80%, up to about 75%, up to about 70%, up to about 65%, up to about 60%, up to about 55%, up to about 50%, up to about 45%, up to about 40%, up to about 35%, up to about 30%, up to about 25%, up to about 20%, up to abut 15%, up to about 10%, up to about 5% by weight of the blend. In some embodiments the amount of sodium in the food product is from about 20% to about 50% less than the amount of sodium in a control food product and the food product and control food product have similar salt taste and similar flavor. A further aspect of the present invention relates to a composition comprising: (i) a blend of sodium chloride and potassium chloride; and (ii) trehalose at an amount of less than 1.5% by weight of the composition. In one embodiment, the blend comprises up to about 99.9% by weight potassium chloride. In other embodiments, the blend comprises up to about 95%, up to about 90%, up to about 85%, up to about 80%, up to about 75%, up to about 70%, up to about 65%, up to about 60%, up to about 55%, up to about 50%, up to about 45%, up to about 40%, up to about 35%, up to about 30%, up to about 25%, up to about 20%, up to about 15%, up to about 10%, or up to about 5% by weight potassium chloride. In another embodiment, the blend comprises about 50% by weight sodium chloride and about 50% by weight potassium chloride. In an example of this embodiment, the amount of trehalose is about 0.20% by weight of the composition. In a further embodiment, the blend comprises 5% by weight sodium chloride and 95% by weight potassium chloride. In an example of this embodiment, the amount of trehalose is about 0.95% by weight of the composition. Another aspect of the present invention relates to a composition comprising: (i) sodium chloride; (ii) potassium chloride; and (iii) trehalose at a sufficient amount to obtain a composition having less bitterness and/or metallic flavor than a control composition that does not contain trehalose. CGL03/0470WO

In one embodiment, the bitterness and/or metallic flavor is attributed to potassium chloride.

"Flavor" refers to a taste sensation or a blend of taste and smell sensations evoked by a

substance in the mouth.

Another aspect of the present invention relates to a food product comprising:

(i) one or more food ingredient(s); and

(ii) a blend of trehalose and sodium chloride and potassium chloride, wherein the blend

provides an amount of trehalose that is less than 1.5% by weight of the food product. The

amount of trehalose in the blend may vary depending upon the amount of the blend in the

food product. In one embodiment, the amount of trehalose in the blend is calculated by the

formula:

[S + 1.49] * T = 149,

wherein S refers to the amount of sodium chloride generally found in the food product, as a

percent by weight of the food product, and T refers to the percent by weight of the trehalose in

the salt substitute used in place of the sodium chloride, or "S".

Non-limiting examples of this embodiment include those set forth in Table I below:

TABLE I

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According to one embodiment, if a finished food product typically included 1% sodium chloride (i.e. the food product is 1% sodium chloride, 99% other food ingredients), that 1% sodium chloride could be replaced by 1% by weight of a blend, which blend includes 59.9% trehalose and 40.1% of a combination of sodium chloride and potassium chloride. According to another embodiment, if a finished food product typically included 1% sodium chloride, that 1% sodium chloride could be replaced by trehalose and 1% by weight of a blend comprising sodium chloride and potassium chloride (i.e. the original 1% sodium chloride would be replace by more than 1% of a composition comprising trehalose, sodium chloride, and potassium chloride). In another embodiment, the present invention relates to a food composition, comprising: a blend of trehalose and sodium chloride, wherein the food composition includes less than 1.5% trehalose by weight of the food product. The amount of trehalose in the blend can be far greater than 1.5% (w/w) of the blend, for example up to 99% (w/w) of the blend so long as the amount of trehalose in the food product is less than 1.5% by weight of the food product. In another embodiment, the present invention relates to a food composition, comprising: a blend of trehalose and potassium chloride, wherein the food composition includes less than 1.5% trehalose by weight of the food product. The amount of trehalose in the blend can be far greater than 1.5% (w/w) of the blend, for example up to 99% (w/w) of the blend so long as the amount of trehalose in the food product is less than 1.5% by weight of the food product. Non-limiting examples of food products include snacks, bouillons, soups, soup mixes, broths, sauces, sauce mixes, dressings, condiments, mayonnaise, ketchup, meats, juices, pickles, and baked products. Another aspect of the present invention relates to a food product comprising: (i) one or more food ingredient(s); (ii) sodium chloride; and (iii) trehalose in an amount sufficient to potentiate the salty flavor contributed by the sodium chloride to the food product without affecting the food product's sweetness, wherein the salty flavor is comparable to that of a control food, the control food product contains more CGL03/0470WO

sodium chloride than the food product, and the control food product does not contain trehalose. Except for the noted differences, a "control food product" is otherwise substantially the same as the food product. As an example, the food product may have at least about 50%, at least about 45%, at least about 40%, at least about 35%, at least about 30%, at least about 25%, at least about 20%, at least about 15%, at least about 10%, or at least about 10% less sodium chloride than the control product. As another example, the food product is very low sodium, low sodium, light in sodium, lightly salted, or les sodium. "Very low sodium" means 35 mg or less per serving or, if the serving is 30 grams or less or 2 tablespoons or less, 35 mg or less per 50 g of the food product. "Low sodium" means 140 mg or less per serving or, if the serving is 30 g or less or 2 tablespoons or less, 140 mg or less per 50 g of the food product. "Light in sodium" means at least 50 percent less sodium per serving than average reference amount for same food product with no sodium reduction. "Lightly salted" means at least 50 percent less sodium per serving than reference amount. "Less sodium" means at least 25 percent less per serving than reference food. See Publication No. FDA 95-2284. In further embodiments, the food product comprises no more than about 500 mg, no more than about 450 mg, no more than about 400 mg, no more than about 350 mg, no more than about 300 mg, no more than about 250 mg, no more than about 200 mg, no more than about 140 mg, no more than about 100 mg, no more than about 50 mg, or no more than about 35 mg of sodium chloride. Another aspect of the present invention relates to a food product comprising: (i) one or more food ingredient(s); (ii) sodium chloride; and (iii) trehalose at a sufficient amount to obtain a food product having more savory flavor than a control product that does not contain trehalose. Non-limiting examples of savory flavors include spicy flavor, vegetable flavor, tomato flavor, dill flavor, meat flavor, poultry flavor, chicken flavor and reaction flavors that are added or generated during heating of the food product. In one embodiment, the amount of trehalose does not affect the food product's sweetness. CGL03/0470WO

Another aspect of the present invention relates to a food product comprising: (i) one or more food ingredient(s); (ii) sodium chloride; (iii) potassium chloride; and (iv) trehalose at a sufficient amount to obtain a food product having more savory flavor than a control product that does not contain trehalose. In one embodiment, the amount of trehalose does not affect the food product's sweetness. Another aspect of the present invention relates to a food product comprising: (i) one or more food ingredient(s); (ii) sodium chloride; (iii) potassium chloride; and (iv) trehalose at a sufficient amount to obtain a food product having less bitterness and/or metallic flavor than a control product that does not contain trehalose. Another aspect of the present invention relates to a method of preparing a food product, which comprises adding to one or more food ingredient(s), sodium chloride and trehalose at an amount of less than 1.5% by weight of the food product. In one embodiment, the method further comprises adding potassium chloride to the one or more food ingredient(s). Another aspect of the present invention relates to a method of preparing a food product, which comprises adding to one or more food ingredient(s), sodium chloride and trehalose at a sufficient amount to obtain a food product having more savory flavor than a control product that does not contain trehalose. In one embodiment, the amount of trehalose does not affect the food product's sweetness. Another aspect of the present invention relates to a method of preparing a food product, which comprises adding to one or more food ingredient(s), sodium chloride and trehalose at a sufficient amount to impart a salty flavor to the food product without affecting the food product's sweetness, wherein the salty flavor is comparable to that of a control food, the control food product contains more sodium chloride than the food product, and the control food product does not contain trehalose. In one embodiment, the method further comprises adding potassium chloride to the one or more food ingredient(s). CGL03/0470WO

Another aspect of the present invention relates to a method of preparing a food product, which comprises combining one or more food ingredient(s), sodium chloride, potassium chloride and trehalose at a sufficient amount to obtain a food product having less bitterness and/or metallic flavor than a control product that does not contain trehalose, without affecting the food product's sweetness. Another aspect of the present invention relates to a method of preparing a seasoning composition, which comprises combining sodium chloride with trehalose at a sufficient amount to obtain a food product having more savory flavor than a control product that does not contain trehalose. In one embodiment, the amount of trehalose does not affect the food product's sweetness. Another aspect of the present invention relates to a method of preparing a seasoning composition, which comprises combining sodium chloride with trehalose at a sufficient to impart a salty flavor to the food product without affecting the food product's sweetness, wherein the salty flavor is comparable to that of a control food, the control food product contains more sodium chloride than the food product, and the control food product does not contain trehalose. Another aspect of the present invention relates to a method of preparing a seasoning composition, which comprises combining sodium chloride, potassium chloride and trehalose at a sufficient amount to obtain a seasoning composition that has less bitterness and/or metallic flavor than a control composition that does not contain trehalose. In one embodiment, the bitterness and/or metallic flavor is attributed to the potassium chloride. The above-described salt-substitutes can be applied to a broad range of food products. Similarly, the sodium content of a broad range of food products can be reduced (as compared to the typical sodium content of the food product) while still maintaining desirable saltiness and flavor by making the food products with sodium and trehalose, and optionally potassium. The food products include those identified as "suitable food products" above. It will be apparent to one of ordinary skill in the art that specific embodiments of the present invention may be directed to one, some or all of the above-indicated aspects as well as other aspects, and may encompass one, some or all of the above- and below- indicated embodiments, as well as other embodiments. Thus, for example, a food product in CGL03/0470WO

accordance with the present invention may comprise: i) one or more food ingredients, ii) potassium chloride; and, iii) trehalose in an amount sufficient to mask the bitterness of the potassium chloride, and/or in an amount sufficient to obtain a food product having more savory flavor than a control product that does not contain trehalose, and/or in an amount sufficient to enhance the salty flavor of potassium chloride. Other than in the working examples, or where otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified by the term "about". Accordingly, unless indicated to the contrary, such numbers are approximations that may vary depending upon the-desired properties sought to be obtained by the present invention. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should be construed in light of the number of significant digits and ordinary rounding techniques. While the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the working examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

EXAMPLES

The following examples are illustrative of the present invention and are not intended to be limitations thereon. EXAMPLE 1: CREAMY CHICKEN SOUP Laboratory Method of Preparation 1. Weigh about a third of the water in a stainless steel container. 2. Blend in the water at medium speed (~7500rpm), the chicken base using a high-speed mixer such as Omni International's PowerGen Homogenizer. 3. Add and blend in Trehalose, potassium chloride and salt mixture at medium speed. 4. Add and blend in all remaining dry ingredients - starches, maltodextrin, onion and garlic powders, beta-carotene and white pepper. CGL03/0470WO

5. Melt the mono- and di-glyceπdes in the soybean oil and blend. Blend in the oil at high

speed (~ 10,000 rpm).

6. Add remaining water and blend at low speed, ~ 5000 rpm, to a uniform consistency.

7. Transfer the soup preparation into Mason jars and cook the samples in a pressure

cooker at 15 psi for 40 minutes. Follow the recommended method for pressure-

cooking. After the samples cooled down, store them in a refrigerator for evaluations.

Example IA: Control 50 % Reduced Sodium

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Example IB: Creamy Chicken Soup - 50 % Sodium Reduction

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Example 1C: Creamy Chicken Soup - 50 % Sodium Reduction

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Example ID: Cream Chicken Sou - 50 % Sodium Reduction

EXAMPLE2: REGULARPOPCORN

Example 2A: Control - Regular Popcorn Ingredients Percent Weight (g) Popcorn Kernels 65.20 41.1 Butter Oil 32.35 20.40 Butter Flavor # 816.32/EN (FONA) 0.10 0.06 Salt (Flour Salt) 2.35 1.48 100.00 ( 63.06 Popcorn Serving Size = 36 grams Sodium Content = 1.48 g x 23 /58.5 x 36/63.06 g x 1000 mg = 332 mg Na /Serving Size Example 2B: Salt Blends for Sodium Reduction in Regular Popcorns (Use level or equivalent saltiness of salt blends per 100 g of popcorns = 1.48 g / 0.76 = 1.95 grams) CGL03/0470WO

Example 2C:

Compositions of Reduced Sodium Regular Popcorn Formulations

Example 2D:

Reduced Sodium Salt Blend Form' Control - Light Popcorn Ingredients Percent Weight (g) Popcorn Kernels 85.74 60.02 Butter Oil 11.37 7.96 Butter Flavor # 816.32/EN (FONA) 0.10 0.07 Salt (Flour Salt) 2.79 1.95 100.00 70.00 Popcorn Serving Size = 36 grams Sodium Content = 1.95 g x 23 /58.5 x 36/70.0 g x 1000 mg = 394 mg Na /Serving Size CGL03/0470WO

Example 2E:

Salt Blends for Sodium Reduction in Light Popcorns

(Use level or equivalent saltiness of salt blends per 100 g of popcorns = 1.95 g / 076 = 2.57 grams)

Example 2F:

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EXAMPLE 3: FRIED AND BAKED POTATO CHIPS

Compositions of Reduced Sodium Salt Blends

(Use level or equivalent saltiness of salt blends per 100 g of potato chips = 1.60 g / 0.76 =

2.10 grams)

Lab Preparation of Reduced Sodium Potato Chips

1. Weigh 100 grams of unsalted fried potato chips 2. Weigh the salt (1.60 g) or salt blend (2.10 g) into a salt shaker 3. Transfer the chips into a confectionery type coating vessel lined with a plastic bag. 4. While rotating the vessel at low speed, heat the chips with a heat gun for about 45 seconds or when tiny oil droplet become visible on the chips. 5. Shake the salt or salt blend uniformly on the relatively hot chips while the vessel was rotating. 6. Remove and package the chips in suitable storage bags for evaluations.

EXAMPLE 4. SODIUM REDUCTION IN A PROCESSED CHEESE

Processed cheese formulations produced in the lab and evaluated by a group of tasters

are shown in Tables below.

Table 4.1. Processed Cheddar Cheese Spread Produced with only salt.

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Table 4.2. 25 % Reduced Sodium Processed Cheddar Cheese Spread

With Trehalose Without Trehalose

Laboratory Process:

1. Weigh the milks into a stainless steel container placed in a water bath. While agitating the milk at medium speed, 8,000 rpm, using a PowerGen Homogenizer, heat the milk to about 120 0F. 2. Blend in the warm milk the carrageenan. Continue the heating and the agitation. 3. Increase the mixing speed to 10,000 rpm and blend in the whey protein concentrate and salt. (If Trehalose and KCl are present, blend in the Trehalose followed by KCl before adding and blending in the salt). 4. Add and blend in the Cheddar cheese flavor and then the annatto. 5. Continue the agitation and heat the processed cheese preparation to 185 °F (85 0C). 6. Hold the temperature of the cheese preparation at 185 0F for 5 minutes to Pasteurize the sample. 7. Pour the hot cheese preparation into suitable containers. Cover the cheese spread and store the samples in a refrigerator.

Product Evaluations

The reduced sodium sample in Table 4.2 was evaluated against the control sample

shown in Table 4.1. The taste test results are summarized as followed.

The 25 % reduced sodium samples were evaluated first by 8 people in the lab and later

by 9 panelists at a round table. Both taste panels described the 25 % reduced sodium cheese

sample with Trehalose as being quite close to the control in saltiness perception and overall CGL03/0470WO

flavor. The cheese flavor was described as being balanced when compared to the sample that did not contain Trehalose.

EXAMPLE 5: SENSORY RESULTS FOR BBQ POTATO CHIPS, CHICKEN BREAST, AND CHILL

Samples were analyzed in a round table discussion sensory panel. Products were tasted one at a time and discussed prior to tasting the next sample. At the conclusion of tasting each product type the overall conclusions were discussed and agreed upon. Products discussed here are chicken breast, BBQ potato chips and chili. The samples were as follows CGL03/0470WO

BBO Potato Chips (three samples)

1. Control 2. 50% reduced sodium (1.6:1, 1:1) 3. 50% reduced sodium (1.6:1, 0:1)

50% Salt Reduction (#2 above):

100.000% 100.000% 100.000% 100.000% Procedure for coating potato chips: 1 - Place potato chips in drum coater. 2 - Heat chips to 120F with heat gun. 3 - Add preblended dry spice blend topically to potato chips until uniformly coated. 4 - Remove chips from coater and package. CGL03/0470WO

Chicken Breast

Control

60% reduced sodium (1.6:1, 1:1)

60% reduced sodium (1.6:1, 0:1)

100.00% 100.00% 100.00% Vacuum Marinade Process: 1 - Dissolve phosphate in cold water first with high speed agitation. 2 - Add salt, KCl, Trehalose and ice, mix thoroughly until ice is melted. 3 - Place refrigerated whole muscle chicken breasts into a vacuum tumbler. 4 - Add water, phosphate, salt, KCl, Trehalose solution to vacuum tumbler. 5 - Place top on vacuum tumbler and draw vacuum to about 16 inches of mercury. Close vacuum and tumble chicken and solution under vacuum for 30 minutes at 9 RPM. 6 - Stop tumbler, release vacuum, remove chicken. 7 - Cook chicken...see below.

Cooking Process: 1 - Heat oven to 350F. 2 - Place marinaded chicken breasts on cooking tray and cover with aluminum foil. 3 - Bake chicken until thickest part of chicken breast is 165F. 3 - Remove chicken from oven and cool to 4OF. 4 - Cut into l"xl" cubes, package and freeze. 5 - Reheat to 165F for evaluation. CGL03/0470WO

Chili Control

50% reduced sodium (1.6:1, 1:1)

50% reduced sodium (1.6:1, 0:1)

100.00% 100.00% 100.00%

Processing: 1 - Slurry water and meat in a cooking vessel until meat is thoroughly mixed. 2 - Add the remaining ingredients and mix thoroughly until uniformly mixed. 3 - With continuous stirring or swept surface agitation, cook mixture to 195F and hold for 10 minutes. 4 - Cool mixture to 4OF. 5 - Portion chili in Zip Lock Freezer bags and freeze. 6 - KEEP FROZEN until ready to use.

Preparation: 1 - Remove frozen chili from package and place into sauce pan. 2 - Heat chili to 165F and serve CGL03/0470WO

Method of Analysis: Samples were analyzed in a round table discussion sensory panel. Samples were assigned randomized three digit codes and analyzed individually by panelists (6 panelists for chicken breast, 8 panelists for processed cheese spread, 10 panelists for BBQ potato chips, and 8 panelists for chili). Panelists were provided with common descriptors for each product type and were asked to taste the samples and describe the characteristics of each sample on an individual ballot. Products were tasted one at a time and discussed prior to tasting the next sample. Panelists were instructed to use a water-Sprite- water rinse between each sample. At the conclusion of tasting each product type the overall conclusions were discussed and agreed upon. Panelists were asked to focus on the saltiness and were then instructed to focus on individual attributes. Panelists were given a break to rest their palette between tasting each sample type. Conclusions:

The chicken breast products tasted were determined to have equivalent saltiness, therefore, the control, the product with trehalose, and the product without trehalose all had the equivalent salt performance. Panelists were split on overall liking of the control product and the product without trehalose, however, the product with trehalose was least preferred.

The salt performance of the BBQ potato chips was deemed to be equivalent in all three products. However, the overall flavor of the sample with trehalose was most similar to the control. The flavor of the sample without trehalose was distinctly different than the other samples.

The salt performance of the chili samples was deemed to be equivalent in all three products. The flavor of the sample without trehalose was more similar to the control than the sample with trehalose. CGL03/0470WO

EXAMPLE 6. EFFECT OF TREHALOSE IN REDUCED SODIUM PRODUCTS CONTAINING SODIUM AND POTASSIUM CHLORIDE

Example 6.1. Preliminary Taste Tests in Finished Food Products Based on informal taste tests in the laboratory by six tasters, the addition of trehalose (0.1% to 2% w/w) was found to improve the flavor and/or mask the off-flavor of reduced sodium products containing sodium and potassium chlorides. Table Salt. A salt solution (1.5% w/w) was made using Morton® Lite Salt® (a mixture of sodium chloride, potassium chloride, iodide and a freeflowing agent, with 50% less sodium than regular table salt). To one part of this solution, about 0.5% w/w trehalose was added to make a first sample, and about 0.8% w/w trehalose was added to make a second sample. The solutions with and without trehalose were evaluated using sensory tests for taste. The Lite Salt® with trehalose was found to taste better than the Lite Salt® without trehalose. Impact of Sweetness on Bitterness. Sugar was used as the sweetener and its level was adjusted in such a way that it had the same sweetness as Trehalose. Using Morton® Lite Salt®, a 1.5% (w/v) salt solution was made. Sucrose was added to this solution at a 0.5% level. The salt solution with Trehalose was compared against a control Morton® Lite Salt® solution. The taste panel (4 tasters) felt that the product with sugar tasted as bad as or worse than the product without sugar. Tasters described the taste as "dirty" compared to the metallic taste of the control. Soup. Campbell's regular cream of chicken soup and low sodium level cream of chicken soup (Campbell's® Healthy Request® soup, which contained both sodium chloride and potassium chloride, with an advertised 30% less sodium than Campbell's® regular soups) was used for testing. Two different levels of Trehalsoe were added to the low sodium product — 0.5% and 0.8% weight percent. Two tasters (food scientists) evaluated the above products against a control of low sodium soup without the addition of trehalose. Both tasters found that the soups with the addition of trehalose tasted better (masked the off taste of potassium chloride) than the commercial low sodium soup. Bouillon Cubes. Soup was made from both regular and low sodium Herbox brand chicken cubes in accordance with package directions. Trehalose was added to the chicken CGL03/0470WO

broth at 0.5% weight percent levels. The two tasters found that the low sodium soup with Trehalose tasted much better than low sodium soup without Trehalose. The off taste of potassium (from potassium chloride added to low sodium soups) did not come through during the tasting of the product with Trehalose. Broth. Swanson broth was used to evaluate if Trehalose enhanced the saltiness of the broth. Trehalose was added to the chicken broth at 0.5% and 0.8% (by weight) to see if there was an increased saltiness perception. The two tasters did not find any significant increase in saltiness perception compared to the standard product. Example 6.2: Reduced Sodium Brines with Trehalose. Brines (2.25% w/w salt) containing sodium and potassium chlorides were made in the laboratory for tasting to see the effect of trehalose addition. Different levels of sodium reductions were tried out using potassium chloride so that saltiness is maintained. Potassium chloride levels varied from 50% to 95% of the salt blends, and the corresponding sodium chloride levels were 50% to 5%. Trehalose levels ranged from 0.2% of the salt blend for 50% salt blends to 0.95% of the salt blend for 95% potassium chloride blends. As the level of trehalose was increased for a particular salt blend (50/50 or 80/20) from low to high, tasters observed that bitterness/off-taste reached a minimum level and any level of trehalose beyond that did not have any effect of bitterness. On the other hand, saltiness, as perceived by tasters (measured by taste scores), went from low to high then to low again before reaching an optimum and falling down again. For example in 50/50 salt blends, the first high score for saltiness (acceptability) occurred at very low levels of trehalose (0.2%) and then saltiness (acceptability) perception fell down followed by optimum score for saltiness at 1.5% trehalose. At higher levels of trehalose beyond 1.5%, sweetness interfered with saltiness and the perception of saltiness was compromised. Methodology: Different levels of Trehalose were added to a salt solution that contained both Sodium Chloride and Potassium Chloride. In the initial tests, Trehalose was added to the solution at various levels to find the optimum level that would mask the metallic taste of potassium. Samples were compared against Salt (NaCl) solution and Lite Salt (NaCl & KCl) solution at the same concentration. 2 experimenters did the evaluations. CGL03/0470WO

Materials: Standard salt from stores were used in the experimentation (Morton's Salt or Cargill Evaporated Salt) unless otherwise mentioned. Anhydrous Potassium Chloride was obtained from Mallinckrodt Chemical Company. Trehalose used in the experiment was the standard product sold by Cargill.

Test 1: Determine the level of Trehalose in 50% Reduced Salt Solutions: Used a 2.25% (w/v) salt or low salt mixture (KCl & NaCl) solution was used for evaluation. Evaluation: 2.25% solutions of the above mixtures were compared against Control salt solution and Lite Salt (50% KCl & NaCl) solutions. Final Trehalose concentration in the finished salt blend solutions: 0.011%, 0.017%, 0.006%, 0.0008%, & 0.0045% respectively from Blends 1 to 5. Three tasters tasted the salt solutions for bitterness and saltiness against the two controls. Level of 0.15% Trehalose did not mask the bitter taste of potassium chloride. Salt blends with 0.2% & 0.25% Trehalose masked the bitterness and was similar to the control salt solution (only NaCl). Test 2: Determine Trehalose level in 80% Reduced Salt Solutions Methodology: Replaced 80% NaCl with Potassium Chloride and tested the following levels of Trehalose in the 2.25% (w/v) salt blend solution: 0.009%, 0.0124%, & 0.016%. These products were evaluated against 2.25% (w/v) standard salt solution and also salt solution that had 80% KCl & 20% NaCl Evaluation: Of the sample salt solutions with both KCl (80%) & NaCl (20%), the salt solution with 0.009% Trehalose in the finished salt solution tasted the best with respect to least bitterness and closest in taste to control salt (NaCl) solution. CGL03/0470WO

Test 3: Determine Trehalose level in 95% Reduced Salt Solutions Methodology: Similar to the previous method. Replaced 95% of NaCl with KCl. Trehalose levels in the finished salt blend solutions (2.25% w/v) were as follows: 0.0135%, 0.018% and 0.024% Evaluation: Even at high levels of Trehalose (0.024%), bitterness was not as pronounced as the control salt solution that had 95% KCl and 5% NaCl (without Trehalose) but at these levels of substitution, the taste of the Lite salt solution (KCl & NaCl) was changed and was different from control salt solution (100% NaCl). Test 4: Determination of Trehalose levels for 60% reduction of sodium Methodology: Similar to above methods. Replaced 60% of NaCl with KCl. The Trehalose levels in the finished salt blend solutions (2.25%w/v) were: 0.0057%, 0.0063%, 0.0074% & 0.0086%. Evaluation: 0.0063% in the finished product tasted the best with respect to both masking bitterness and similar tasting to salt solution (only NaCl). Test 5: Determination of Trehalose levels for 65% reduction of sodium Methodology: Similar to the above. Replaced 65% of NaCl with KCl. The Trehalose levels in the finished salt blend solutions (2.25%w/v) were: 0.0052%, 0.0063%, 0.0074% & 0.0086%. Evaluation: 0.0074% in the finished product tasted the best with respect to both masking bitterness and similar tasting to salt solution (only NaCl). Test 6: Determination of Trehalose levels for 70% reduction of sodium Methodology: Similar to the above. Replaced 70% of NaCI with KCl. The Trehalose levels in the finished salt blend solutions (2.25 %w/v) were: 0.0077%, 0.0081%, 0.0086% & 0.009%. Evaluation: 0.0086% in the finished product tasted the best with respect to both masking bitterness and similar tasting to salt solution (only NaCl). Test 7: Effect of Trehalose % on saltiness and bitterness in 66% Reduced Salt Brines: Methodology: In our previous trials, we had tested intermediate levels of Trehalose (0.0052 to 0.086%) in the finished product and found that 0.0074% Trehalose gave the best tasting low sodium brine solutions at 65% reduction in sodium. In our next trials, we reduced CGL03/0470WO

the salt concentration of the salt blend brines to 1.15% since tasters described the previous

concentration as too salty, and also the sensitivity of the tasters was improving. The amount

of salt blend mixed with water was adjusted so that all solutions had the same level of sodium

and potassium chlorides. Some of the levels were repeated since it was difficult to perceive

bitterness/off taste of the solutions and also there were some differences in comments among

the tasters.

Evaluation: Three tasters who were involved in evaluating the products from the

beginning of the project did the evaluation & tasting. It was very difficult to evaluate such

minor differences in the levels of Trehalose especially in salt brine solutions. Hence, it was

decided to do further sensory work using chicken broth rather than salt brine.

TABLE 6.1

EXAMPLE 7. TREHALOSE IN CHICKEN BROTH

Methodology: Prepared a no salt chicken broth from fresh chicken; salt blends were

used at 0.76% of the chicken broth.

Procedure for Making Chicken Broth: 9 to 10 Ib of cut chicken and 6 quarts of cold

water was brought to boil in a pot. Heat was reduced to simmer and the contents simmered

for another 30 minutes. The fat layer was removed from the top. To this mixture, we added:

10 celery ribs, 1 bay leaf, 1 cup chopped onion, 1 cup chopped carrots, 12 sprigs of parsley.

The pot was continued to simmer for another 2.5 hours with lid partially covered. The

contents strained, and refrigerated. Before serving, any fat separating on top was removed. CGL03/0470WO

66% reduced sodium blends using potassium chloride was used to test the

effectiveness of Trehalose at four different levels in the finished chicken broth; 0%, 0.0019%,

0.0025% & 0.0031%.

Evaluation: The low sodium chicken broth with 0.0025% had the best overall taste

with respect to saltiness and absence of bitterness. Below this level, bitterness was perceived

and above that level, salt taste was compromised and tasters described the flavor as not

natural.

Example 7(a) Further Studies in Chicken Broth:

Tasters found that it was easier to perceive the differences in Trehalose treatments in

chicken broth and hence we did a range of Trehalose % in the finished chicken broth. It was

decided to increase the salt (low sodium blend) level to 1.5% to detect small differences.

Specifically, various levels of trehalose were tested in 50/50 salt blends (KCl and NaCl), and

it was found that 1.5% of trehalose in the salt blend had the best overall taste. The results are

presented in Table 7.1 and Table 7.2, below.

TABLE 7.1

CGL03/0470WO

TABLE 7.2

Sucrose was also tested as another control at reduced levels to provide the same

sweetness as trehalose to verify if the bitterness masking and enhancement of saltiness is

observed. It was found that there was some level of bitterness masking but saltiness was

compromised leading to an off-taste even at very low levels of sucrose.

Example 7(b) Sensory Testing in Chicken Broth:

We wanted to confirm the results observed by the technical team and wanted more

people to do the sensory testing of the products. Chicken broth was made as described above.

A panel of 12 testers evaluated three chicken broths. Tasters tested all three samples at the

same time. Specfically, a panel of 12 taste testers evaluated three chicken soup broths, a

standard formula, a standard formula with 50% reduction in NaCl using KCl and a standard

formula with 50% reduction in NaCl using KCl and trehalose. The broths were presented in

random order. The panelists rated the samples for overall liking, liking of the flavor,

bitterness intensity and saltiness intensity.

The sample of "50% reduction in NaCl using KCl" had a significantly (p<0.05) higher

bitterness intensity score than the other two samples did. There were no other significant

differences between the other products. CGL03/0470WO

The results from ANOVA on the scores are provided in Table 7.3, below.

TABLE 7.3

Attribute 100% NaCl 50% reduction 50% reduction in NaCI using in NaCi using KCl KCl and trehalose

Overall Liking1 6.33 6.25 7.17 0.1479 Flavor Liking2 6.50 6.33 6.58 0.8912 Bitterness 5.92b 11.61a 6.43b 0.0272 Intensity2 Saltiness 33.43 38.53 31.48 0.2941 Intensity

1 1 = "Dislike Extremely"; 2 = "Dislike Very Much"; 3 = "Dislike Moderately"; 4 = "Dislike

Slightly"; 5 = "Neither Like nor Dislike"; 6 = "Like Slightly"; 7 = "Like Moderately", 8 =

"Like Very Much"; 9 = "Like Extremely"

2 1.4 = "barely detectable"; 6.1 = "weak"; 17.2 = "moderate"; 35.4 = "strong"; 53.3 = "very

strong"; 100 = "strongest imaginable"

EXAMPLE 8. The inventors have found that trehalose can have one or more effects

in salty and/or savory products, including: (A) improved flavor of low sodium food products

that had both sodium and potassium chlorides at very low levels of trehalose (e.g. less than

0.05% in the finished food products); (B) enhanced perception of saltiness at very low levels

of trehalose (e.g. less than 0.05% in the finished food products); and (C) enhanced flavor

perception of certain savory flavors in food products that had regular or reduced sodium

levels.

Example 8.1. Effect of Trehalose in Enhancing Saltiness

The effect of saltiness enhancement was studied using very low levels of trehalose to

high levels of trehalose. Initial findings indicate that trehalose at 0.0225% in the finished CGL03/0470WO

product (equivalent of 1.5% in salt blend) enhanced the saltiness the most and the saltiness perception showed a pattern (low to high to low to optimum to low) similar to the study with salt blend (sodium and potassium chlorides). In the next set of trials, the level of enhancement due to trehalose in chicken broth was evaluated by comparing chicken broths that had a blend of salt and trehalose against those that had various levels of salt alone. In initial trials, it was found that the level of salt enhancement was close to 20% when trehalose was used along with salt (the product with 1.5% salt blend (salt and trehalose) tasted as salty as that product with 1.8% salt alone). Example 8.2: Effect of Trehalose in Enhancing Savory Character In studies with chicken broth, tomato juice (contained both sodium and potassium chlorides), tomato soup, pickles, chicken base (as provided by Five Star Foods), it was found that the addition of trehalose brought out the characterizing flavor of the above products in addition to enhancing saltiness and masking undesirable flavors. The tasters who tasted the various products mentioned a difference in the perception of the overall flavor of the product in terms of savory character. Also, when the products with trehalose were compared to those that had comparable salt, tasters perceived a much smoother delivery of flavor (for example in the dill pickle, the dill flavor in the trehalose added product had less flavor spikes when eating a individual pickle pieces). In initial trials, tasters sampled chicken broth and other savory food products such as tomato juice, chicken base, etc. with trehalose against products without trehalose. The addition of trehalose enhanced the savory flavor of the products (chicken or tomato flavor) when compared to products that did not have trehalose. A low sodium broth with trehalose was also compared against a broth without sodium reduction. It was found that the savory character of the broth with trehalose was better than that of the corresponding product without trehalose. The optimum level of trehalose that showed this effect was below 0.05% and the flavor enhancing effect varied with the level of trehalose. CGL03/0470WO

EXAMPLE 9. OTHER FOOD PRODUCTS

Example 9.1 Dili Pickles.

Dill Pickle was made using brines with regular and reduced sodium brines. The

products with reduced sodium (50% reduction) and trehalose scored as well as products with

regular levels of sodium and the product without trehalose had an off-flavor.

Methodology: In this experiment Dill Pickles were made with salt and 50% Reduced

sodium blends with and without Trehalose. Trehalose was used at levels of 0.015% in the

brine used for pickling. The formula for the brine & also the ratio of cucumber to brine are

given below. The products were packed in glass jars and pasteurized at 165F.

Evaluation: Four tasters evaluated the product. Three tasters chose the Trehalose

containing product, as the best tasting product and one person liked the control best. All the

four tasters described the taste of the product without Trehalose as having a bitter/off taste.

Example 9.2 Lo-Carb Dry Mix Soup:

Methodology: One trial of a Dry Potato Soup Mix was used as a model system to

further study the effect of Trehalose. The following salt & salt blends were used in the mix

for evaluation: 80% Reduced Sodium Blend (80g KCl, 2Og NaCI blend with 0.4g Trehalose),

50% Reduced Sodium Blend (5Og NaCl, 5Og KCl with 0.2g Trehalose), Morton Lite Salt

(50% NaCl & 50% KCl). CGL03/0470WO

Lo Carb Dry Mix was made up of the following ingredients:

In the above formula, salt was replaced by the salt blends & Morton lite salt. Evaluation (3 tasters): 50% Reduced salt blend tasted the best in overall taste with respect to least bitterness and most salty taste. Example 9.3. Potato Chips: Methodology: In order to disperse the salt 1-2 gm of salt or salt blend in 100 gm of chips, we mixed with the salt with a carrier such as maltodextrin. Two different DE (dextrose equivalent with differing sweetness) of maltodextrins were tried out - 18 & 7.5 DE. We found that 7.5 DE was the acceptable type of maltodextrin for this application. Trehalose levels in the salt blends were as follows: 0.2 g of Trehalose/5 Ogm of KCl/50 gm of NaCl for 50% Reduced sodium sample, 0.33 gm of Trehalose/65 gm of KCl/35 gm of NaCl, 0.95gm of Trehalose/95 gm of KCl/5 gm of NaCl. The finished Trehalose percentages in chips were as follows:0. 004% for 50% reduction, 0.007% for 65% reduction in sodium and 0.019% for 95% reduction of sodium.

0.5 gm of salt blend was added to 2.5 gm of 7.5 DE maltodextrin. This mixture was added to 50 gm of unsalted potato chips in a Ziploc plastic bag & thoroughly shaken for the seasoning blend to mix. In all the following tests, low sodium salt blends were made using potassium chloride (1 part of sodium chloride was replaced by 1 part of potassium chloride on a mass basis). Unsalted potato chips were salted with low sodium salt blends with and without Trehalose. Evaluation (done by 4 tasters): 50% Reduction of sodium: It was difficult to distinguish between the chips salted with low sodium blends with trehalose and without trehalose since the products without trehalose did not have the off taste of potassium due to the high fat CGL03/0470WO

65% Reduction in sodium: The tasters could not perceive differences in the taste

between the potato chips salted with low sodium blends with and without trehalose. It was

decided to increase the salt reduction levels.

95% Reduction in sodium: The tasters perceived an off taste in both the chips with

and without Trehalose but the chips with trehalose tasted saltier & had a lesser off taste than

the product that did not have trehalose.

Example 9.4. Seasoning Blends:

Methodology: A seasoning blend was created for use in snack products. The formula

for such a blend is given below:

We used the above seasoning blend to do trials of 80% Sodium reduction in potato

chips. In the place of salt in the above formula, we used the low sodium blend with 80% KCl

and 20% NaCl and 0.4 g of Trehalose. 1 gm of seasoning blend was mixed with 3 gm of 7.5

DE maltodextrin (carrier) & the combined mixture was added to 50 gm of unsalted potato

chips (bought from the stores). A control without the Trehalose (low sodium blend without

Trehalose) was also made for comparing against test product. The level of Trehalose in the

finished food product was 0.008% CGL03/0470WO

Evaluation: The tasters (four in number) perceived the off taste in the product without trehalose while the product with trehalose did not have such an off taste. However, the salty taste was faint. Example 9.5. Low Sodium Application in Freshly Made Potato Chips: Methodology: Same seasoning blend was used & the seasoning was applied to freshly made potato chips from potatoes (slice potatoes in 1/16 inch thickness, heat in Frymax vegetable oil at 330F till the slices become brown). Four trials were run; 80% reduction in sodium (80% KCl & 20% NaCl) with and without Trehalose and 95% reduction in sodium with and without Trehalose. Evaluation: The 80% reduced sodium product with trehalose was the best tasting product without any bitterness and good salty taste. The products without trehalose had noticeable bitterness especially the product with 95% reduction in sodium without trehalose. The 95% reduced sodium product with trehalose had some bitterness. Example 9.6. Sensory Testing in Potato Chips: Methodology: Later experiments in potato chips were conducted using "No Salt Added" chips from the market. Chips with No Salt added were fed in to a coating drum, and chips were heated by directing hot air from the dryer to the outside of the drum. When chips get warm and the oil droplets start appearing on the surface of the chips, salt (or salt blend) is added using a shaker uniformly over all the chips. Specifically, consumer reaction to reduced-sodium potato chips was measured on six different chip samples. Samples were presented monadically over a six day-period. Participants picked up their chip samples at 10:00 am, consumed them at their leisure and recorded their results by 5:00 pm the same day. The table below gives some summary statistics for the results. Letter superscripts in the Overall liking results are the LSD groupings. Samples A and B had significantly (p<0.05) higher overall liking scores than samples D and E did. The difference between samples B and C is not statistically significant at the 95% level, but is at the 90% level. This provides some evidence that Trehalose provides some benefit in terms of overall liking at the 33% sodium reduction level. CGL03/0470WO

EXAMPLE lO: TOMATO SOUP; Many of the tomato products in the market have a large amount of salt and we wanted to test our technology in a tomato-based soup. Methodology: We developed a formula that is similar to some of the tomato soups that are available in the market. The formula for the Tomato Soup Control is given below:

3 1="Dis!ike Extremely"; 2="Dislike Very Much"; 3="Dislike Moderately"; 4="Dislike Slightly"; 5="Neither Like nor Dislike"; 6="Like Slightly"; 7="Like Moderately", 8="Like Very Much"; 9="Like Extremely" 4 Percent in top two boxes on a 5-point scale. 5 The sample was an acceptable substitute for my normal brand (%) CGL03/0470WO

50% reduced sodium tomato soup was made using a mixture of sodium and potassium

chloride (0.5% each) and in the 50% reduced sodium soup with Trehalose, the level of

Trehalose used was 0.015%. The products were processed using pressure cooker at 15 psi for

lO min.

The following levels of Trehalose percentages in the finished product were tried;

Control with salt, 0.0033%, .01%, 0.015%, 0.031%, 0.053%, 0.11%, & 0.25%. The amounts

of salt blend were adjusted so that the salt blend level was the same across all the treatment

(to compensate for the various levels of Trehalose in the salt blend).

Evaluation: Three experienced tasters tasted the products and the top two products in

terms of overall liking was those products that had final Trehalose levels of 0.0033% &

0.015% respectively. Trehalose level at 0.053% had a good score in overall liking by two

tasters but the saltiness was compromised.

Sensory Testing of Tomato Soup: Further testing was done with tomato soup by

consumers (Cargill employees). The panelists tasted all the three soups (control, 50% reduced

sodium product without Trehalose and 50% reduced product with Trehalose) at the same CGL03/0470WO

time. The soups were rated on a 9-point hedonic scale6. The soups were presented in randomized order and served at comparable temperatures. Analysis of variance was used to test for differences between the samples, and the results are shown below. Overall Liking Summary

*Means with the same letter are not significantly different. (p<0.05) As shown the sample with NaCl scored significantly higher than the other two samples. None of the samples had particularly high scores. Comments: The results showed that the there was no significant difference between the low sodium product with and without Trehalose. The products were tasted by project team members (with experience in tasting low sodium products) and only thing the team members found was that the taste of the low sodium tomato soup with trehalose was different from the control (100% salt) product in it's flavor also. The Trehalose seemed to have affected not only the potassium chloride taste but also the tomato soup profile of the product. When tasters tasted all the three products (control and two samples with lower sodium content), they found that the product with Trehalose was more different from control than product without Trehalose. This seemed to have affected the scores also.

6 1="Dislike Extremely"; 2="Dislike Very Much"; 3="Dislike Moderately"; 4="Dislike Slightly"; 5="Neither Like nor Dislike"; 6="Like Slightly"; 7="Like Moderately", 8="Like Very Much"; 9="Like Extremely" CGL03/0470WO

EXAMPLE 11. CHICKEN SOUP:

Methodology: We developed a chicken soup formula for testing by consumers using a formal monadic (sensory testing of only one soup at a time) testing. The formulas tested are set forth in Example 1. The table below gives the summary statistics for overall liking, flavor liking and saltiness liking. Summary Liking Statistics

Analysis of variance with Tukey's honest significant difference (HSD) revealed the following significant differences between the treatments. Flavor Liking: Low Trehalose significantly higher (better) than No Trehalose (p<0.05). Saltiness Liking: Low Trehalose significantly higher (better) than No Trehalose (p<0.05). Salt Control significantly higher (better) than No Trehalose (0.05<p<0.10) No other significant differences were found. Appendix 2 contains histograms of the overall liking scores for each treatment. The table below gives the flavor JAR scores for all samples. Note that the percent of panelists that felt that the broth had too little flavor was relatively low for Wixon. CGL03/0470WO

Flavor JAR Scores

The table below gives the saltiness JAR scores for all samples. Note that the JAR profiles are very similar for all five samples, and there were no significant differences in saltiness JAR scores between the treatments. Saltiness JAR Scores

There were no significant differences between the treatments with respect to aftertaste and bitterness scores as shown in the tables below. CGL03/0470WO

Aftertaste Scores

Bitterness Scores

EXAMPLE 12. ENHANCEMENT OF SALTINESS USING TREHALOSE;

Initial Salt Brine Tests:

We also studied the effect of saltiness enhancement of Trehalose. In our initial

studies, we used salt brine at 1.12% salt. The Trehalose levels were varied as below:

We first wanted to find out what level of Trehalose has the maximum impact on

saltiness & then compare this data with different controls (only salt without Trehalose) to

understand how much enhancement occurs using Trehalose. CGL03/0470WO

Chicken Broth Test:

Methodology: Chicken Broth was made as mentioned above. Trehalose was added at

different levels. The salt amount was adjusted for the addition of Trehalose so that same level

(1.5% Salt) was added to all the samples.

100% NaCl % Trehalose Finished Product Salty taste Bitter Taste O Offff T Taste Overall Liking Salt 6 0 0 0.003 7 0 0 0.008 5 0 0 0.015 6 0 0 9 0.0225 10 0 0 11 0.05 4 0 6 3 0.075 6 0 3 7 0.1 4 0 4 5 0.5 3 0 4 5 1.0 4 0 5 3 1.5 • 3 0 6 3 3.0 3 0 8 2 10.0 2 0 Ii 2

Evaluation: Four tasters tasted the samples & the chicken broth with 0.0225%

Trehalose had the best overall taste and also increased the saltiness much more than any other

levels without sacrificing any other desirable taste properties.

Determination of the Trehalose enhancement of Saltiness:

Methodology: We took the best-case scenario of salt enhancement in Chicken Broth

(0.0225% of Trehalose) compared to different levels of salt controls. The salt level of chicken

broth studied was 1.5% and this salt level with Trehalose (0.0225%) was compared (using CGL03/0470WO

taste tests) against higher levels of salt without Trehalose to understand the level of enhancement. The salt levels used were 1.8%, 1.9% and 2% salt. Evaluation: Two tasters tasted the product and found that chicken broth with 1.5% salt with Trehalose (0.0225%) tasted similar to 1.8% salt without any Trehalose. The invention being thus described, it will be apparent to those skilled in the art that the same may be varied in many ways without departing from the spirit and scope of the invention. Such variations are included within the scope of the invention to be claimed