Use of hyaluronic acid and salts thereof for taste enhancement

Abstract

The application discloses a use of hyaluronic acid and its salt in taste enhancement, and belongs to the field of salt and sugar reduction. The application discloses a use of hyaluronic acid, hyaluronic acid salt and a mixture of hyaluronic acid and hyaluronic acid salt in taste enhancement, wherein the hyaluronic acid salt includes sodium salt and the like, and the molecular weight is 80.4 kDa-1090 kDa. Based on the use, the intake of condiments or taste compounds in condiments of an animal body, especially a human body, can be reduced under the condition of obtaining the same taste feeling, or a more lasting feeling of taste can be obtained under the condition of intaking the same amount of condiments or taste compounds.

Description

TECHNICAL FIELD

[0001] The present application relates to the use of hyaluronic acid and its salts in the enhancement of taste, and belongs to the field of salt and sugar reduction. BACKGROUND

[0002] Taste is one of the important physiological sensations of the human body, and largely determines the choice of food by people. Taste plays an important role in the regulation of food intake, body nutrition and metabolic regulation. The receptor of taste is taste bud, which is mainly distributed on the surface of the tongue and the lingual margin, and also has scattered distribution on the surface of the oral and pharyngeal mucosa. Taste bud is composed of taste cells, which have taste receptors on the cells and can detect and distinguish various tastes. Taste cells have no axon, but are surrounded by sensory nerve endings, and an axon connection is formed between the two, which is activated by neurotransmitters released by taste cells to produce nerve impulses, which are transmitted to the central nervous system to cause taste.

[0003] Flavoring, condiment, seasoning, refers to the color, aroma, taste of dishes, promote appetite, and is beneficial to human health of auxiliary food. Its main function is to improve the quality of dishes, meet the taste needs of consumers, so as to stimulate appetite and improve human health. In a broad sense, flavoring includes salty, sour, sweet, umami and spicy agents, such as salt, soy sauce, vinegar, monosodium glutamate, sugar, star anise, fennel, Sichuan pepper, mustard, etc. However, excessive intake of flavoring, such as salt and sugar, is an important risk factor for cardiovascular and cerebrovascular damage, such as hypertension, myocardial hypertrophy, atherosclerosis, stroke, etc. Controlling the intake of salt, sugar and other substances has been recognized worldwide as one of the most cost-effective chronic disease intervention strategies. For example, using low-sodium salt or other mineral salt, salty / umami flavor enhancer (such as amino acid, peptide, organic acid and aromatic compound) and salty peptide, or a combination of the foregoing to reduce salt intake. For example, using non-nutritive high-potency sweeteners, sugar alcohols and dietary fibers to achieve sugar reduction.

[0004] Of course, reducing the intake of flavoring should ultimately aim to not reduce the perception of taste. A variety of natural and non-natural compositions and / or compounds have been added to food, beverages and / or edible (eatable) compositions to improve and enhance the taste of salt, umami, sweet, bitter, spicy and sour, etc., thereby reducing the addition and intake of corresponding flavoring. For example, a small amount of "guide" compound connected with heteroaryl is reported to be able to regulate T1R1 / T1R3 "salty and umami" taste receptors, thereby reducing salt and increasing saltiness.

[0005] Hyaluronic acid is a natural high molecular linear polysaccharide composed of D-glucuronic acid and N-acetylglucosamine disaccharide units connected repeatedly. Due to its unique molecular structure and physicochemical properties, it has many important physiological functions, such as moisturizing, lubricating, promoting wound healing, anti-inflammatory, etc. SUMMARY

[0006] The present application discloses the use of hyaluronic acid, hyaluronic acid salts, and mixtures of hyaluronic acid and hyaluronic acid salts in taste enhancement, wherein the hyaluronic acid is a natural high molecular linear polysaccharide formed by repeating connection of disaccharide units composed of D-glucuronic acid and N-acetylglucosamine, the hyaluronic acid salts include sodium salt, potassium salt, calcium salt, magnesium salt, or zinc salt, etc., the molecular weight of the hyaluronic acid is 80.4 kDa to 1090 kDa (preferably 100 kDa to 400 kDa), and the taste enhancement refers to enhancing the taste sensation of animals, especially humans, to taste, including salty, fresh, sweet, spicy, bitter, and sour. The taste enhancement can also be expressed as taste promotion, taste regulation, and taste enhancement.

[0007] The present application discloses the use of hyaluronic acid, hyaluronic acid salts, and mixtures of hyaluronic acid and hyaluronic acid salts in taste enhancement, based on which the intake of seasonings or taste compounds in food and seasonings can be reduced in animals, especially humans, under the condition of obtaining the same taste sensation; and based on which more persistent taste sensation can be obtained in animals, especially humans, under the condition of ingesting the same amount of seasonings or taste compounds. The seasonings refer to substances capable of providing animals or humans with taste or taste sensation, containing taste compounds capable of providing animals or humans with any one or two or more than two taste sensation of salty, sour, sweet, fresh, bitter, and spicy. The seasonings include table salt, soy sauce, sauce, vinegar, monosodium glutamate, chicken essence seasoning, sugar, seasoning products, etc. The soy sauce includes brewed soy sauce and prepared soy sauce. The sauce includes sweet bean sauce, yellow sauce, bean paste, raw sauce, and bimodal chili sauce. The vinegar includes brewed vinegar and prepared vinegar. The monosodium glutamate includes sodium glutamate. The seasoning products include solid seasoning (such as seafood powder seasoning, livestock and poultry powder seasoning, etc.), semi-solid seasoning (such as peanut butter, sesame paste, mayonnaise, salad dressing, mustard sauce, hot pot seasoning, and chili oil, etc.), liquid seasoning (such as chicken juice seasoning, barbecue sauce, oyster sauce, fish sauce, pickling liquid, seasoning wine, etc.), and edible seasoning oil (such as pepper oil, mustard oil, chili oil, and blended sesame oil, etc.). The food includes substances edible or drinkable by humans, such as meat products, beverages, instant food, frozen drinks, quick-frozen food, biscuits, cans, puffed food, potato food, candy products, jelly, wine, preserved fruits, jam, pickled vegetables, pastries, flavored fish products, surimi products, salted aquatic products, dried aquatic products, reprocessed egg products, etc. The food also includes special medical purpose formula food and pet food.

[0008] The present application discloses the use of hyaluronic acid, hyaluronic acid salts, and mixtures of hyaluronic acid and hyaluronic acid salts in the enhancement of taste, the implementation of which includes adding hyaluronic acid, hyaluronic acid salts, or mixtures of hyaluronic acid and hyaluronic acid salts to the raw materials of food, during the preparation of food, or during the consumption of food. For example, hyaluronic acid, hyaluronic acid salts, or mixtures of hyaluronic acid and hyaluronic acid salts, and seasonings are added in sequence or simultaneously to the raw materials of food, during the preparation of food, or during the consumption of food; or hyaluronic acid, hyaluronic acid salts, or mixtures of hyaluronic acid and hyaluronic acid salts are used as components of seasonings to make seasonings, which are added to the raw materials of food, during the preparation of food, or during the consumption of food. The food includes substances for human consumption or drinking, such as meat products, beverages, instant foods, frozen drinks, quick-frozen foods, biscuits, cans, puffed foods, potato foods, candy products, jellies, wine, preserves, jams, pickles, pastries, flavored fish products, surimi products, salted aquatic products, dried aquatic products, reconstituted egg products, and the like. The food also includes special medical purpose formula foods and pet foods.

[0009] The present application discloses the use of hyaluronic acid, hyaluronic acid salts, and mixtures of hyaluronic acid and hyaluronic acid salts in the enhancement of taste, for the enhancement of salty taste, which can enhance the salty taste by about 25-250%, i.e., reduce the amount of salty agents by 20-60%, such as reducing the amount of sodium chloride by 20-60%; for the enhancement of sweet taste, which can enhance the sweet taste by about 50-300%, and reduce the amount of sweet agents by about 30-70%, such as reducing the amount of sucralose by 30-70%; for the enhancement of spicy taste, which can enhance the spicy taste by about 25-250%, i.e., reduce the amount of spicy agents by 20-60%, such as reducing the amount of chili by 20-60%; for the enhancement of bitter taste, which can enhance the bitter taste by about 25-250%, i.e., reduce the amount of bitter taste substances (such as coffee) by 20-60%; for the enhancement of sour taste, which can enhance the sour taste by about 25-250%, i.e., reduce the amount of sour substances (such as citric acid) by 20-60%.

[0010] The present invention discloses the use of hyaluronic acid, hyaluronic acid salts, and mixtures of hyaluronic acid and hyaluronic acid salts for taste enhancement. The hyaluronic acid or hyaluronic acid salt forming hyaluronic acid has a molecular weight of 80.4 kDa to 1090 kDa (preferably 100 kDa to 400 kDa). The amount of hyaluronic acid, hyaluronic acid salt, or mixture of hyaluronic acid and hyaluronic acid salt can be determined based on the upper limit allowed by food-related laws and regulations and the characteristics (including taste, solubility, and viscosity) required to be presented by the food itself, for example, 0.1 to 25% of the mass of the prepared flavored food. For example, the hyaluronic acid has a molecular weight of 100 kDa and an amount of 0.2% of the total mass of the prepared salty food, the hyaluronic acid has a molecular weight of 100 kDa and an amount of 0.4% of the total mass of the prepared salty food, the hyaluronic acid has a molecular weight of 400 kDa and an amount of 0.2% of the total mass of the prepared salty food, or the hyaluronic acid has a molecular weight of 400 kDa and an amount of 0.4% of the total mass of the prepared salty food.

[0011] The present invention discloses the use of hyaluronic acid, hyaluronic acid salts, and mixtures of hyaluronic acid and hyaluronic acid salts for taste enhancement. The hyaluronic acid or hyaluronic acid salt forming hyaluronic acid has a molecular weight of 80.4 kDa to 1090 kDa (preferably 100 kDa to 400 kDa). The amount of hyaluronic acid, hyaluronic acid salt, or mixture of hyaluronic acid and hyaluronic acid salt can be determined based on the upper limit allowed by food-related laws and regulations and the characteristics (including taste, solubility, and viscosity) required to be presented by the food itself, for example, 0.1 to 25% of the mass of the prepared flavored food. For example, the hyaluronic acid has a molecular weight of 100 kDa and an amount of 0.2% of the total mass of the prepared salty food, the hyaluronic acid has a molecular weight of 100 kDa and an amount of 0.4% of the total mass of the prepared salty food, the hyaluronic acid has a molecular weight of 400 kDa and an amount of 0.2% of the total mass of the prepared salty food, or the hyaluronic acid has a molecular weight of 400 kDa and an amount of 0.4% of the total mass of the prepared salty food.

[0012] The present invention discloses the use of hyaluronic acid, hyaluronic acid salts, and mixtures of hyaluronic acid and hyaluronic acid salts for taste enhancement. The hyaluronic acid or hyaluronic acid salt forming hyaluronic acid has a molecular weight of 80.4 kDa to 1090 kDa (preferably 100 kDa to 400 kDa). The amount of hyaluronic acid, hyaluronic acid salt, or mixture of hyaluronic acid and hyaluronic acid salt can be determined based on the upper limit allowed by food-related laws and regulations and the characteristics (including taste, solubility, and viscosity) required to be presented by the food itself, for example, 0.1 to 25% of the mass of the prepared flavored food. For example, the hyaluronic acid has a molecular weight of 100 kDa and an amount of 0.2% of the total mass of the prepared salty food, the hyaluronic acid has a molecular weight of 100 kDa and an amount of 0.4% of the total mass of the prepared salty food, the hyaluronic acid has a molecular weight of 400 kDa and an amount of 0.2% of the total mass of the prepared salty food, or the hyaluronic acid has a molecular weight of 400 kDa and an amount of 0.4% of the total mass of the prepared salty food.

[0013] The present invention discloses the use of hyaluronic acid, hyaluronic acid salts, and mixtures of hyaluronic acid and hyaluronic acid salts for taste enhancement, based on which food products can be designed and prepared that reduce the amount of taste compounds in a seasoning or a seasoning. The food products include substances that can be consumed or drunk by humans. For example, meat products, beverages, instant foods, frozen beverages, quick-frozen foods, biscuits, cans, puffed foods, potato foods, confectionery products, jellies, wine, candied fruits, jams, chutneys, pastries, flavored fish products, surimi products, salted aquatic products, dried aquatic products, reconstituted egg products, and the like. The food products also include special medical use formula foods and pet foods.

[0014] The present invention relates to hyaluronic acid or its salts and their use as taste enhancers, taste potentiators, taste modulators, taste intensifiers, taste enhancers, umami taste enhancers, or salty taste enhancers, or sweet taste enhancers, or spicy taste enhancers, or sour taste enhancers, or bitter taste enhancers, and or taste enhancers, and their use. Based on this use, hyaluronic acid or its salts can be combined with food or beverages or sauce seasonings or dishes or edible food compositions or pharmaceutical compositions (in particular, pharmaceutical preparations with special flavors such as honey, peach, brown sugar, lemon, and other sweet and sour flavors for children's medicines or women's medicines) in different molecular weights and different concentration orders. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 is the saltiness intensification effect of hyaluronic acid in 50 mM NaCl at different molecular weights (100 kDa and 400 kDa) and concentrations (0.2% and 0.4%).

[0016] Figure 2 is the intensification effect of 0.2% hyaluronic acid at different molecular weights (100 kDa, 400 kDa, and 1090 kDa) on different salt concentration solutions (10, 30, 50, 70, 90 mM NaCl).

[0017] Figure 3 is the effect of hyaluronic acid at different molecular weights (100 kDa and 400 kDa) and concentrations (0.2% and 0.4%) on saltiness prolongation.

[0018] Figure 4 is the effect of hyaluronic acid at different molecular weights (100 kDa and 400 kDa) and concentrations (0.2% and 0.4%) on saltiness over time.

[0019] Figure 5 is the change in chip adsorption mass and thickness of physical coating and chemical modification.

[0020] Figure 6 is the scanning electron microscope image of the chemically modified mucin layer and the acetic acid buffer solution after flushing.

[0021] Figure 7 is a physical coated mucin layer and acetic acid buffered solution rinse followed by scanning electron microscopy.

[0022] Figure 8 is the change in frequency (f) and dissipation (D) values for 100 kDa 0.2% hyaluronan.

[0023] Figure 9 is the change in frequency (f) and dissipation (D) values for 100 kDa 0.2% hyaluronan with Na added.

[0024] Figure 10 is the change in frequency (f) and dissipation (D) values for 100 kDa 0.4% hyaluronan.

[0025] Figure 11 is the change in frequency (f) and dissipation (D) values for 100 kDa 0.4% hyaluronan with Na added.

[0026] Figure 12 is the change in frequency (f) and dissipation (D) values for 400 kDa 0.2% hyaluronan.

[0027] Figure 13 is the change in frequency (f) and dissipation (D) values for 400 kDa 0.2% hyaluronan with Na added.

[0028] Figure 14 is the change in frequency (f) and dissipation (D) values for 400 kDa 0.4% hyaluronan.

[0029] Figure 15 is the change in frequency (f) and dissipation (D) values for 400 kDa 0.4% hyaluronan with Na added.

[0030] Figure 16 is the change in release of sodium fluorescein over time for different polysaccharides (100 kDa 0.2% hyaluronan and 0.3% gum arabic) in artificial tongue gels.

[0031] Figure 17 is the shear curve for hyaluronan of different molecular weights (8 kDa, 100 kDa, 400 kDa and 1090 kDa).

[0032] Figure 18 is the sweetness enhancement effect of hyaluronan of different molecular weights (100 kDa and 400 kDa) and concentrations (0.2% and 0.4%) in 43.9 μΜ sucralose.

[0033] Figure 19The enhancement effect of hyaluronic acid with different molecular weights (100kDa and 400kDa) on solutions with different sugar concentrations (5.5, 11, 22, 43.9, 87.8, 175.6, 351.2, 702.5 μM sucralose) is shown.

[0034] Figure 20 The effects of hyaluronic acid with different molecular weights (100kDa and 400kDa) and concentrations (0.2% and 0.4%) on the duration of sweetness.

[0035] Figure 21 The effect of hyaluronic acid with different molecular weights (100kDa and 400kDa) and concentrations (0.2% and 0.4%) on sweetness over time.

[0036] Figure 22 The effects of hyaluronic acid with different molecular weights (100kDa and 400kDa) and concentrations (0.2% and 0.4%) on the taste perception of hot and salty sauce.

[0037] Figure 23 The effects of hyaluronic acid with different molecular weights (100kDa and 400kDa) and concentrations (0.2% and 0.4%) on the taste perception of sweet chili sauce.

[0038] Figure 24 The effects of hyaluronic acid with different molecular weights (100kDa and 400kDa) and concentrations (0.2% and 0.4%) on the taste perception of pickled pepper sauce.

[0039] Figure 25 The effects of hyaluronic acid with different molecular weights (100kDa and 400kDa) and concentrations (0.2% and 0.4%) on the perception of (bitter) taste in coffee.

[0040] Figure 26 The effects of hyaluronic acid with different molecular weights (100kDa and 400kDa) and concentrations (0.2% and 0.4%) on the perception of the (acid) taste of coffee.

[0041] Figure 27 It is the interaction between hyaluronic acid of different molecular weights (8kDa, 100kDa and 400kDa) and mucin. Detailed Implementation

[0042] In this application, "seasoning" means a substance that enables an animal or human to perceive taste or flavor, containing flavor compounds that enable an animal or human to perceive one or more of the following tastes: salty, sour, sweet, umami, bitter, and spicy.

[0043] The "taste-imparting compound" in the present application means a substance contained in a substance ingested into the oral cavity of an animal or a human being, which imparts any one or two or more of salty taste, sour taste, sweet taste, umami taste, bitter taste, and pungent taste to the sensory organ (tongue, taste bud).

[0044] The "taste-enhancing" in the present application means enhancing the sense of taste of an animal body, particularly a human body, by taste buds, for example, the sense of salty, umami, sweet, pungent, bitter, and sour by taste buds in a human body. The "taste-enhancing" in the present application can also be expressed as taste promotion, taste adjustment, taste reinforcement. The "taste-enhancing" can be sensed by a human being or detected by an instrument, for example, an instrument for detecting brain signal reflex. The "taste-enhancing" includes allowing a human being to have a stronger or more remarkable sense of taste, allowing a human being to have a more persistent sense of taste in time, allowing a taste-imparting compound to diffuse or permeate more widely and persistently in the oral cavity, and allowing a taste-imparting compound to stay more persistently in the oral cavity.

[0045] In one embodiment of the present application, the hyaluronic acid refers to a natural high-molecular linear polysaccharide formed by repeating connection of disaccharide units composed of D-glucuronic acid and N-acetylglucosamine, and food-grade hyaluronic acid having a molecular weight of 80.4 kDa to 1090 kDa (preferably 100 kDa to 400 kDa) and salts thereof. The preferred molecular weight range or molecular weight is 100 kDa to 400 kDa, or 100 kDa, or 400 kDa.

[0046] In one embodiment of the present application, the hyaluronic acid salt is a sodium salt, a potassium salt, a calcium salt, a magnesium salt, or a zinc salt.

[0047] In some embodiments of the present application, the hyaluronic acid, the hyaluronic acid salt, or a mixture of the hyaluronic acid and the hyaluronic acid salt is added to a raw material of a food, in the process of making a food, or in the process of eating a food, thereby reducing the intake amount of a seasoning or a taste-imparting compound in the seasoning. As an example of the food, there are meat products, dairy products, beverages, instant foods, frozen drinks, quick-frozen foods, biscuits, cans, puffed foods, potato foods, confectionery products, jellies, wines, preserves, jams, chutneys, cakes, flavored fish products, surimi products, salted aquatic products, dried aquatic products, ordinary foods such as reconstituted egg products, pet foods, and special medical use formula foods.

[0048] In some embodiments of the present application, hyaluronic acid, a salt of hyaluronic acid, or a mixture of hyaluronic acid and a salt of hyaluronic acid is used as a component of a seasoning to make a seasoning, which is then added to a foodstuff during the preparation of the foodstuff, during the preparation of the foodstuff, or during the consumption of the foodstuff. These seasonings contain known natural and / or artificial taste compounds acceptable for use in foodstuffs, including any one or two or more of salty, umami, sweet, bitter, sour, and bitter taste compounds. Exemplary seasonings include table salt, chicken bouillon, sugar, brewed soy sauce, prepared soy sauce, sweet bean sauce, yellow bean sauce, doubanjiang, raw sauce, two-peak chili sauce, brewed vinegar, prepared vinegar, monosodium glutamate, seafood powder seasoning, poultry powder seasoning, peanut butter, sesame paste, mayonnaise, salad dressing, mustard sauce, hot pot seasoning, chili oil, chicken juice seasoning, barbecue sauce, oyster sauce, fish sauce, pickling liquid, seasoning wine, pepper oil, mustard oil, chili oil, and blended sesame oil.

[0049] In some embodiments of the present application, when hyaluronic acid, a salt of hyaluronic acid, or a mixture of hyaluronic acid and a salt of hyaluronic acid is used to enhance salty taste, it can enhance salty taste by about 25-250%, i.e., reduce the amount of salty taste agent by 20-60%, such as reducing the amount of sodium chloride by 20-60%; when it is used to enhance sweet taste, it can enhance sweet taste by about 50-300%, and reduce the amount of sweet taste agent by about 30-70%, such as reducing the amount of sucralose by 30-70%; when it is used to enhance spicy taste, it can enhance spicy taste by about 25-250%, i.e., reduce the amount of spicy taste agent by 25-60%, such as reducing the amount of chili pepper by 20-60%; when it is used to enhance bitter taste, it can enhance bitter taste by about 25-250%, i.e., reduce the amount of bitter taste agent (such as coffee) by 20-60%; and when it is used to enhance sour taste, it can enhance sour taste by about 25-250%, i.e., reduce the amount of sour taste agent (such as citric acid) by 20-60%. The enhancement of salty taste by about 25-250% means that the addition of 1 part of a taste compound or seasoning can achieve the same salty taste as the addition of 1.25-2.5 parts of the taste compound or seasoning. The reduction of the amount of salty taste agent by 20-60% means that the amount of the taste compound or seasoning is reduced by 20-60% to achieve the same salty taste. The enhancement of sweet taste, spicy taste, bitter taste, and sour taste, and the reduction of the amount of the taste agent are understood accordingly.

[0050] In some embodiments of the present application, the amount of hyaluronic acid, a salt of hyaluronic acid, or a mixture of hyaluronic acid and a salt of hyaluronic acid used in a seasoned foodstuff is 0.1-25% of the total mass of the foodstuff.

[0051] In some embodiments of the present application, hyaluronic acid, a salt of hyaluronic acid, or a mixture of hyaluronic acid and a salt of hyaluronic acid, is also mixed with other known or future discovered taste enhancers (e.g., compounds reported to modulate the T1R1 / T1R3 "umami" taste receptor) to form a taste enhancer composition that contains a substance that facilitates the storage stability of hyaluronic acid and / or its salt, the taste enhancing effect, and / or the incorporation into a food product, such as a prepared meal, a milk tea, and a vegetarian soup mix, etc.

[0052] Example 1 Use of hyaluronic acid, a salt of hyaluronic acid in enhancing saltiness

[0053] 1. Take 1000 mL of water, and slowly add 8 g of hyaluronic acid (or sodium hyaluronate) with a molecular weight of 100 kDa to prepare a 0.8% mass concentration of hyaluronic acid solution (or sodium hyaluronate solution). Take another 1000 mL of water, and add 5.85 g of NaCl to prepare a 100 mM NaCl solution. Mix the hyaluronic acid solution (or sodium hyaluronate solution) and the NaCl solution at a weight ratio of 1:1 to prepare a 50 mM NaCl solution containing 0.4% mass concentration of hyaluronic acid with a molecular weight of 100 kDa.

[0054] 2. Prepare a 50 mM NaCl solution containing 0.4% mass concentration of hyaluronic acid with a molecular weight of 400 kDa in the same way.

[0055] 3. Take 1000 mL of water, and slowly add 4 g of hyaluronic acid (or sodium hyaluronate) with a molecular weight of 100 kDa to prepare a 0.4% mass concentration of hyaluronic acid solution (or sodium hyaluronate solution). Take another 1000 mL of water, and add 5.85 g of NaCl to prepare a 100 mM NaCl solution. Mix the hyaluronic acid solution and the NaCl solution at a weight ratio of 1:1 to finally prepare a 50 mM NaCl solution containing 0.2% mass concentration of hyaluronic acid with a molecular weight of 100 kDa.

[0056] 4. Prepare a 50 mM NaCl solution containing 0.2% mass concentration of hyaluronic acid with a molecular weight of 400 kDa in the same way.

[0057] 5. Take 2000 mL of water, and add 5.85 g of NaCl to prepare a 50 mM NaCl solution.

[0058] Sensory evaluation method: 15 professional sensory evaluators taste the solutions obtained in the foregoing ①-⑤, 10 mL of the solution obtained in the foregoing ①-⑤ is placed in a 30 mL sensory evaluation cup, and is randomly presented to the sensory evaluators. The sensory evaluators spit out any solution in ①-④ after holding it in the mouth for 10 s, then rinse their mouths with clean water for 2 min, and then taste the next solution, and then compare it with solution ⑤ (50 mM of pure NaCl solution). The saltiness of the solutions obtained in ①-④ is defined as: None: no saltiness difference; +: slightly saltier; ++: saltier; +++: very salty.

[0059] By Figure 1 As can be seen from the experimental results shown in the table, the 50 mM NaCl solution added with hyaluronic acid or sodium salt thereof having a molecular weight of 100 kDa or 400 kDa has higher saltiness than the solution containing only the same concentration of NaCl. Specifically: (1) when the concentrations of hyaluronic acid or sodium salt thereof are the same, the 100 kDa molecular weight hyaluronic acid has a better effect on enhancing saltiness than the 400 kDa molecular weight hyaluronic acid. (2) When the molecular weight of hyaluronic acid is 100 kDa, the 0.2% mass concentration of hyaluronic acid or sodium salt thereof has a better effect on enhancing saltiness than the 0.4% mass concentration of hyaluronic acid. When the molecular weight of hyaluronic acid is 400 kDa, the 0.2% and 0.4% mass concentrations of hyaluronic acid or sodium salt thereof have similar effects on enhancing saltiness, and there is no significant difference.

[0060] Example 2 Use of hyaluronic acid and hyaluronic acid salt in saltiness enhancement (saltiness-scores curve quantification)

[0061] ① Respectively weigh 1.2, 3.6, 6, 8.4, 10.8 g of NaCl, and add 1000 g of water to prepare 20, 60, 100, 140, and 180 mM NaCl solutions. Develop a saltiness-score curve: dilute the NaCl solution with water at a weight ratio of 1:1 to prepare 10, 30, 50, 70, and 90 mM NaCl solutions, and assign scores to them as 10 mM (1.0 point), 30 mM (3.0 points), 50 mM (5.0 points), 70 mM (7.0 points), and 90 mM (9.0 points).

[0062] ② Respectively weigh 1.2, 3.6, 6, 8.4, 10.8 g of NaCl, and add 1000 mL of water to prepare 20, 60, 100, 140, and 180 mM NaCl solutions. Then, respectively add 1000 mL of 0.4% mass concentration of molecular weight 100 kDa, 400 kDa, 10.9 x 10 5The hyaluronic acid aqueous solution of Da was mixed at a weight ratio of 1:1. The hyaluronic acid NaCl solution was prepared with a hyaluronic acid mass concentration of 0.2%, a molecular weight of 100 kDa, 400 kDa and 1090 kDa respectively, and a NaCl concentration of 10, 30, 50, 70 and 90 mM respectively.

[0063] Sensory evaluation method: 15 professional sensory evaluators were selected and trained. First, taste the saltiness-scoringsolution in ①, and the saltiness scores are 1, 3, 5, 7 and 9 in turn. Through 3-4 times of training, the scoring is mastered. Then taste the NaCl solution with different concentrations of hyaluronic acid with a molecular weight of 100 kDa, 400 kDa and 1090 kDa in ②, and score based on the saltiness-scorings curve. The tasting method is to place the sample in a 30 mL tasting cup, hold the NaCl solution or the hyaluronic acid NaCl solution in the mouth for 10 s, then spit it out and score immediately. Rinse the mouth for 2 min between tasting 2 samples.

[0064] From the experimental results, it can be seen that:

[0065] (1) The addition of low and medium molecular weight hyaluronic acid (such as the hyaluronic acid with a molecular weight of 100 kDa and 400 kDa in this embodiment) has different enhancing effects on the saltiness in the range of 10-90 mM salt concentration.

[0066] (2) Low molecular weight hyaluronic acid has a better enhancing effect on saltiness than medium and low molecular weight hyaluronic acid. High molecular weight hyaluronic acid (such as the hyaluronic acid with a molecular weight of 1090 kDa in this embodiment) has an inhibitory effect on the perception of saltiness.

[0067] Specifically:

[0068] From the sensory evaluation of the different concentrations of NaCl solution added with different molecular weight of hyaluronic acid, it can be seen that the taste of NaCl solution added with low molecular weight of hyaluronic acid (such as 100 kDa of hyaluronic acid in this embodiment) can obtain 40% higher salt taste perception than the same concentration of NaCl solution without adding hyaluronic acid, the taste of NaCl solution added with medium-low molecular weight of hyaluronic acid (such as 400 kDa of hyaluronic acid in this embodiment) can obtain 25% higher salt taste perception than the same concentration of NaCl solution without adding hyaluronic acid, and the taste of NaCl solution added with high molecular weight of hyaluronic acid (such as 1090 kDa of hyaluronic acid in this embodiment) is lower than the same concentration of NaCl solution without adding hyaluronic acid, and the high molecular weight of hyaluronic acid has a certain degree of inhibition on saltiness, because the high molecular weight of hyaluronic acid has too high viscosity, which will produce excessive adhesion on the oral mucosa, resulting in heavy feeling and even obvious after-bitterness and other bad taste, which directly affects the taste perception of taste buds. The experiment on the influence of high molecular weight of hyaluronic acid on taste is not limited to the concentration of hyaluronic acid used in this embodiment, and lower concentration will also have a certain degree of inhibition on different tastes.

[0069] Example 3 Influence of hyaluronic acid concentration on the intensity change of saltiness over time

[0070] ① 1000 mL of water was weighed and 8 g of 100 kDa hyaluronic acid (or sodium hyaluronate) was slowly added to prepare a 0.8% mass concentration of hyaluronic acid solution (or sodium hyaluronate). 1000 g of water was weighed and 5.85 g of NaCl was added to prepare a 100 mM NaCl solution. The hyaluronic acid solution and the NaCl solution were mixed in a weight ratio of 1:1 to prepare a 50 mM NaCl solution containing 0.4% mass concentration of 100 kDa hyaluronic acid.

[0071] ② Refer to the method in ① to prepare a 50 mM NaCl solution containing 0.4% mass concentration of 400 kDa hyaluronic acid.

[0072] ③ 1000 mL of water was weighed and 4 g of 100 kDa hyaluronic acid (or sodium hyaluronate) was slowly added to prepare a 0.4% mass concentration of hyaluronic acid solution (or sodium hyaluronate). In 1000 g of water, 5.85 g of NaCl was added to prepare a 100 mM NaCl solution. The hyaluronic acid solution and the NaCl solution were mixed in a weight ratio of 1:1 to prepare a 50 mM NaCl solution containing 0.2% mass concentration of 100 kDa hyaluronic acid.

[0073] ④ Prepare a 50mM NaCl solution containing 0.2% hyaluronic acid with a molecular weight of 400kDa by referring to the method in ③.

[0074] ⑤ Establish salinity-scoring curves: Dilute NaCl solution with water at a weight ratio of 1:1 to prepare NaCl solutions of 10, 30, 50, 70, and 90 mM, and assign scores as follows: 10 mM corresponds to 1.0 point, 30 mM corresponds to 3.0 point, 50 mM corresponds to 5.0 point, 70 mM corresponds to 7.0 point, and 90 mM corresponds to 9.0 point.

[0075] Sensory evaluation methods:

[0076] The sensory evaluation team consisted of 15 professional sensory evaluators selected and trained. First, they tasted the saltiness-scoring curve solution in ⑤. They practiced this scoring method 3-4 times to become proficient. Then, they evaluated solutions ①-④ one by one. Tasting method: Before tasting the samples, thoroughly cleaned the mouth with 1-2 soda crackers and purified water. Pour the entire solution to be tasted into the mouth and start timing. Stir continuously in the mouth for 5 seconds, then swallow, continuously sensing the saltiness in the mouth and scoring the perceived saltiness according to the saltiness-scoring curve every 1 second until the saltiness disappears. Record the time it takes for the saltiness to disappear from the mouth. The time from when the solution was poured into the mouth to when the saltiness disappeared is the saltiness perception time. After tasting one sample solution, rinse the mouth with water for 2 minutes and rest for 30 seconds before tasting the next sample solution.

[0077] All samples must be tested in three replicates.

[0078] like Figure 3 , Figure 4 As shown, adding low to medium molecular weight hyaluronic acid to NaCl solution enhances the human perception of saltiness, delays the appearance of the saltiness peak, and prolongs the duration of the perceived saltiness peak. Specifically:

[0079] (1) Adding low to medium molecular weight hyaluronic acid (such as hyaluronic acid with molecular weights of 100kDa and 400kDa in this embodiment) can prolong the time of salty taste perception. Compared with not adding hyaluronic acid, adding 0.2% of 100kDa hyaluronic acid can prolong the time of salty taste perception by 27.29%, adding 0.4% of 100kDa hyaluronic acid can prolong the time of salty taste perception by 31.97%, adding 0.2% of 400kDa hyaluronic acid can prolong the time of salty taste perception by 51.16%, and adding 0.4% of 400kDa hyaluronic acid can prolong the time of salty taste perception by 45.16%.

[0080] (2) The effect of adding medium molecular weight hyaluronic acid (such as 400 kDa hyaluronic acid in this example) on prolonging salty taste perception is more obvious than that of adding low molecular weight hyaluronic acid (such as 100 kDa hyaluronic acid in this example). The addition of 100 kDa hyaluronic acid can prolong the salty taste perception by up to 31.97%, while the addition of 400 kDa hyaluronic acid can prolong the salty taste perception by up to 51.16%.

[0081] (3) The effect of adding hyaluronic acid with the same molecular weight but different concentrations on salty taste perception time is not significant.

[0082] Example 4: Study of the effect of hyaluronic acid on the diffusion and retention of Na + (salt taste, salty taste perception) in the oral cavity by dissipative quartz crystal microbalance (QCM-D)

[0083] ① Weigh 100 mL of 100 mM PBS buffer solution with pH 7.4, add 900 mL of ultrapure water to prepare 1000 mL of PBS buffer solution, add 1 M HCl solution to prepare PBS buffer solution with pH 6.7. Weigh 1000 g of the obtained PBS buffer solution with pH 6.7, slowly add 8 g of hyaluronic acid or its salt solution with a molecular weight of 100 kDa to prepare a hyaluronic acid solution with a mass concentration of 0.8%.

[0084] Weigh 1000 mL of PBS buffer solution, add 5.85 g of NaCl to prepare a 100 mM salt solution, and mix the hyaluronic acid solution with a mass concentration of 0.8% with the 100 mM NaCl solution at a weight ratio of 1:1 to prepare a 50 mM NaCl solution containing 0.4% mass concentration of 100 kDa hyaluronic acid.

[0085] ② Refer to the method of ① to prepare a 50 mM NaCl solution containing 0.4% mass concentration of 400 kDa hyaluronic acid, and a 50 mM NaCl solution containing 0.4% mass concentration of 1090 kDa hyaluronic acid.

[0086] ③ Refer to the method of ① to prepare 1000 mL of PBS buffer solution with pH 6.7, slowly add 4 g of hyaluronic acid or its salt solution with a molecular weight of 100 kDa to prepare a hyaluronic acid solution with a mass concentration of 0.8%.

[0087] A 1000 mL of PBS buffer solution with pH 6.7 was prepared according to the method in ①, and 5.85 g of NaCl was added to prepare a 100 mM NaCl solution. The resulting 0.8% hyaluronic acid solution was mixed with the 100 mM NaCl solution at a weight ratio of 1:1 to finally prepare a 50 mM NaCl solution containing 0.2% mass concentration of 100 kDa molecular weight hyaluronic acid.

[0088] IV. A 50 mM NaCl solution containing 0.2% mass concentration of 400 kDa and 0.2% mass concentration of 10.9 x 10 5 Da molecular weight hyaluronic acid was prepared according to the method in ③.

[0089] V. A 1000 mL of PBS buffer solution with pH 6.7 was prepared according to the method in ①, and 6 g of NaCl was weighed and prepared into a 100 mM NaCl solution. It was diluted with water at a weight ratio of 1:1 to prepare a 50 mM NaCl solution.

[0090] VI. A 100 mL of PBS buffer solution with pH 6.7 was prepared according to the method in ①, and 50 mg of Type III porcine gastric mucin (PGM, the most commonly used system for simulating oral mucosa system) was added. It was stirred overnight at 12°C to allow it to fully hydrate. After centrifugation at 800 g for 10 min, the insoluble precipitate was removed to prepare a 0.5 mg / mL mucin solution.

[0091] Dissipation quartz crystal microbalance (QCM-D) is an instrument based on the inverse piezoelectric effect of quartz crystal to detect changes in the mass of its electrode surface. It is the most efficient online tool for analyzing interfacial processes, with nanogram-level sensitivity, and can be used to study interfacial polymer conformation, adsorption kinetics, degradation kinetics, polyelectrolyte layer-by-layer self-assembly, etc. The interaction between hyaluronic acid and mucin was explored by QCM-D technology.

[0092] We determined the binding affinity of the aforementioned different molecular weight hyaluronic acid solutions, as well as the hyaluronic acid and NaCl mixed solution, to the adsorbed mucin layer by dissipation quartz crystal microbalance (QCM-D). QCM-D technology provides two measurement responses: (1) change in resonance frequency (Δf), which represents the change in mass on the chip after adsorption, reflecting the binding affinity between substances, and (2) change in energy dissipation (ΔD), which indicates the thickness and rheological properties of the adsorbed film on the quartz crystal.

[0093] QCM-D measurements were performed with an E4 system from Q-Sense (AB, Gothenburg, Sweden) using gold-coated quartz crystals with a fundamental frequency of 4.95 Hz. Prior to the experiments, the gold-coated quartz crystal chips were cleaned in a UV-ozone cleaner for 10 min to remove surface organic material, followed by a 10 min soak in a 1 : 1 :5 mixture of H2O2(30%), NH 3. H2O (25%) and ultrapure water at 75 °C, followed by rinsing with ultrapure water and ethanol (99%) and drying with N2gas. This cleaning procedure was repeated at least three times. Porcine gastric mucin and hyaluronic acid were attached to the gold-coated quartz crystal chips using a physical coating method and a chemical modification method, respectively. For the physical coating method, the gold-coated quartz crystal chip was rinsed with 10 mM PBS buffer at pH 6.5, and after the baseline was stable (frequency drift of no more than 1 Hz within 30 min), 0.5 mg / mL porcine gastric mucin (PGM) was introduced to form a mucin layer until a stable baseline was reached. PBS buffer was then introduced again for 30 min to rinse off the unbound mucin molecules. Different molecular weights and concentrations of hyaluronic acid solution were then introduced, and after the baseline was stable, PBS buffer was introduced again to rinse off the unbound hyaluronic acid on the chip. For the chemical modification method, the gold-coated quartz crystal chip was rinsed with 10 mM PBS buffer at pH 6.5, and after the baseline was stable, the cleaned gold-coated quartz crystal chip was immersed in ethanol for 20 min at room temperature, then taken out and placed in a 1 mmol MUA ethanol solution for at least 12 hours to form a self-assembled monolayer of thioacids containing carboxylic acid end groups on the gold-coated quartz crystal, obtaining a carboxylic acid functionalized surface. This functionalized surface was labeled as SAM-COOH, then rinsed with ethanol and dried with nitrogen. The chip was installed in the chip cell, and a 200 mM EDC and 50 mM NHS aqueous solution was introduced, followed by ultrapure water and acetic acid buffer (10 mM, pH 4.0) (i). Subsequently, 0.5 mg / mL mucin was added in the acetic acid buffer (ii), followed by rinsing with the acetic acid buffer (iii). Unreacted NHS-esters were inactivated by rinsing with 1 M ETA-HCl in Tris buffer (10 mM, pH 8.5) (iv). Subsequently, the chip was rinsed with PBS buffer (10 mM, pH 6.5) (v). Hyaluronic acid solution (pH 6.5, with or without 100 mM NaCl) was flowed through the mucin layer (vi) to study the interaction. The unbound hyaluronic acid solution was rinsed off with 10 mM PBS (10 mM, pH 6.5) until the baseline was stable (vii). The experimental temperature was 37 °C, and the flow rate was 50 μL / min.

[0094] Scanning electron microscopy was used to observe the microstructure of the mucin layer formed by the physical coating method and the chemical modification method. Prior to the experiments, the silicon wafer was cut into 1 cm x 1 cm squares, and the surface was cleaned with 1 : 1 :5 H2O2(30%), NH3. The cleaned silicon wafer was immersed in a mixture of H2O (25%) and ultrapure water for 10 min, rinsed with ultrapure water and ethanol (99%), and dried with N2gas. The cleaned silicon wafer was then coated with a 15 nm gold palladium layer using a cathode coater.

[0095] Chemically modified silicon wafer: The gold-coated silicon wafer was immersed in absolute ethanol for 20 min, removed and placed in a 1 mmol MUA ethanol solution for 12 h, then rinsed with ethanol and dried with nitrogen gas. The chip was placed vertically in a 200 mM EDC and 50 mM NHS aqueous solution for 20 min, then placed vertically in ultrapure water and acetic acid buffer for 20 min each. It was then placed vertically in a 0.5 mg / mL mucin pH 4 acetic acid buffer solution for 40 min, then placed vertically in a pH 4 acetic acid buffer for 20 min each, and allowed to dry naturally.

[0096] Physically coated silicon wafer: The gold-coated silicon wafer was immersed in absolute ethanol for 20 min, removed and placed in a 1 mmol MUA ethanol solution for 12 h, then rinsed with ethanol and dried with nitrogen gas. The chip was placed vertically in a pH 6.7 PBS buffer solution for 20 min, placed in a 0.5 mg / mL mucin pH 6.7 PBS buffer solution for 40 min, placed vertically in a pH 6.7 PBS buffer solution for 20 min, and allowed to dry naturally.

[0097] Scanning electron microscopy (SEM) was used to observe the dried silicon wafers described above in a field emission scanning electron microscope at a working voltage of 15.0 kV. Micrographs were recorded at a magnification of x500. As shown in Figure 6 , Figure 7 As shown in

[0098] Based on the Voigt-based model, the changes in the adsorbed mass and thickness of the mucin layer when different molecular weights and concentrations of hyaluronic acid solution were passed through the QCM-D chip were measured, as shown in Figure 5 As shown in

[0099] When the NaCl solution was passed through, the resonance frequency and energy dissipation of the chip did not change substantially, indicating that the interaction between the NaCl solution and the mucin layer could be ignored.

[0100] As shown in Figure 8 to Figure 15As shown, when the hyaluronic acid of different molecular weight and different concentration is introduced, the energy dissipation and the resonance frequency both change obviously. It shows that the elasticity of the mucin layer formed on the gold chip surface is weakened and the viscosity is increased, which indicates that the swelling of mucin occurs due to the addition of hyaluronic acid. In addition, the addition of hyaluronic acid reduces the resonance frequency, which indicates that the mass of the gold chip surface is increased, and it shows that the hyaluronic acid adheres to the surface of the mucin layer by interacting with the mucin.

[0101] At the same concentration, with the increase of the molecular weight of hyaluronic acid, the swelling of the mucin layer caused by the addition of hyaluronic acid shows a trend of first increasing and then decreasing. When the hyaluronic acid of 100kDa molecular weight is introduced, the energy dissipation changes the most, and the swelling effect on the mucin layer is the strongest, which indicates that the permeability of the hyaluronic acid of 100kDa molecular weight is the strongest. When the hyaluronic acid of 400kDa molecular weight is introduced, the resonance frequency changes the most, which indicates that the adhesion of the hyaluronic acid of 400kDa molecular weight on the mucin layer is the strongest.

[0102] When the hyaluronic acid of the same molecular weight is introduced, the energy dissipation value changes increases with the increase of the concentration of hyaluronic acid, and the higher the concentration of hyaluronic acid, the stronger the swelling effect on the mucin layer. And the resonance frequency increases, which indicates that the adhesion of the hyaluronic acid on the mucin layer is enhanced, so there are more residues. At low concentration, the permeability and adhesion of 100kDa hyaluronic acid to the mucin layer are better than those of 400kDa hyaluronic acid. With the increase of the concentration, the permeability and adhesion of 400kDa hyaluronic acid to the mucin layer are enhanced, while the permeability and adhesion of 100kDa hyaluronic acid to the mucin layer are weakened. Different concentrations and molecular weights of hyaluronic acid all improve the Na + The adhesion and permeability of the mucin layer.

[0103] When the mixed salt solution of hyaluronic acid is introduced, it can be found that compared with the introduction of NaCl solution only, the energy dissipation and the resonance frequency both change significantly.

[0104] The above experimental results show that the addition of hyaluronic acid causes the permeation of the mucin layer, which can increase the retention of Na + The permeation and diffusion of the mucin layer; and the addition of hyaluronic acid increases the Na + The adhesion on the mucin layer, prolongs the time of Na + Remaining on the mucin layer; and these ultimately affect the perception of the taste buds in the oral mucous layer to Na + The perception of salty taste of food.

[0105] Example 5 detects the influence of hyaluronic acid on the retention and release of Na + (salt taste, salty taste perception) in the oral cavity by using artificial gel tongue

[0106] Mucin was added to polyacrylamide composite hydrogel to simulate the adhesion of hyaluronic acid on the tongue.

[0107] ① 0.02 g of sodium bisulfate (SBS) was dissolved in 16.6 mL of PBS buffer solution, and 1 mg / mL of mucin was added, and dissolved overnight at 4°C. Then 3.4 mL of 30% (29:1) Acr-Bis was added, and 0.2 mL of 10% gel polymerization catalyst ammonium persulfate was added to form a 20 mL artificial tongue solution.

[0108] ② 1000 mL of ultrapure water was weighed, and 0.21 g of sodium bicarbonate, 0.43 g of sodium chloride, 0.75 g of potassium chloride, 0.22 g of calcium chloride dihydrate, 0.91 g of sodium phosphate monobasic monohydrate, and 2.7 g of type III porcine stomach-derived mucin were weighed to prepare an artificial saliva.

[0109] ③ 2 mL of artificial tongue solution was added to a circular glass mold with a diameter of 3 cm, and the gel was placed at room temperature for 24 hours to prepare an artificial gel tongue with a diameter of 3 cm and a height of 0.05 cm.

[0110] ④ Sample solution was prepared, sample A: 0.005 g of fluorescein sodium was dissolved in 100 mL of artificial saliva; sample B: 0.005 g of fluorescein sodium and 0.2 g of 100 kDa hyaluronic acid were dissolved in 100 mL of artificial saliva; sample C: 0.005 g of fluorescein sodium and 0.3 g of gum arabic were dissolved in 100 mL of artificial saliva.

[0111] ⑤ Fluorescein sodium standard solution was prepared: 0.01 g of fluorescein sodium was weighed into 10 mL of artificial saliva in ②, and was diluted to 0.005, 0.00625, 0.008, 0.01, 0.0125, and 0.02 mg / mL of fluorescein sodium standard solution, respectively. The absorbance values of different concentrations of fluorescein sodium standard solution were measured at 485 nm using an ultraviolet spectrophotometer with artificial saliva as the blank solution. The formula for calculating the concentration X of hyaluronic acid and the absorbance value Y was Y = 72.226X + 0.0363, R 2 = 0.9915.

[0112] The artificial gel tongue was soaked in samples A, B, and C prepared in method ④ for 30 min, and then soaked in 10 mL of artificial saliva. Every 1 min, 1 mL of artificial saliva was taken out, and the absorbance value of fluorescein sodium was measured at a wavelength of 485 nm. According to the formula for calculating the concentration of fluorescein sodium and the absorbance value, the content of fluorescein sodium in the artificial saliva every 1 min was calculated.

[0113] As Figure 16As shown, (1) the addition of hyaluronic acid and gum arabic can increase the retention of Na + In the artificial tongue, adhesion and permeation, thereby prolonging the retention of Na + ; (2) the effect of hyaluronic acid on the retention of Na + (salt taste) is better than that of the control group gum arabic.

[0114] Example 6: Study the effect of hyaluronic acid on the retention of Na + (salt taste, salty taste perception) in the oral cavity by rheological study

[0115] ① Take 1000 mL of aqueous solution, slowly add 2 g of hyaluronic acid or its salt solution with a molecular weight of 100 kDa, to prepare a 0.2% mass concentration of hyaluronic acid solution.

[0116] ② According to the same method, respectively prepare 0.2% mass concentration of 400 kDa and 1090 kDa of hyaluronic acid.

[0117] ③ Take 1000 mL of aqueous solution, slowly add 4 g of hyaluronic acid or its salt solution with a molecular weight of 100 kDa, to prepare a 0.4% mass concentration of hyaluronic acid solution, in 1000 mL of aqueous solution, add 5.85 g of NaCl, to prepare a 100 mM salt solution, mix the hyaluronic acid solution with the NaCl solution in a weight ratio of 1:1. Finally, prepare a 50 mM NaCl solution containing 0.2% mass concentration of 100 kDa molecular weight of hyaluronic acid.

[0118] ④ According to the same method, prepare a 50 mM NaCl solution of 0.2% mass concentration of 400 kDa and 1090 kDa molecular weight of hyaluronic acid.

[0119] ⑤ The rheological test of hyaluronic acid solution uses a rotational rheometer. The clamp is a cone plate with an angle of 1° and a diameter of 60 mm. The experimental temperature is controlled by a circulating water bath and a temperature control device, and the controlled temperature is 25°C. The shear rate range of steady shear test is 1 to 1000 s -1 .

[0120] We measured the apparent viscosity of HA with different molecular weights at 50 s -1 , see Figure 17 , the experiment shows that as the relative molecular weight of hyaluronic acid increases, the viscosity of the solution increases. The increase of the viscosity of hyaluronic acid itself changes its flow rate and interfacial behavior in the oral cavity, also changes its characteristics in the adhesion of oral mucosa, further affects the perception of taste substances (such as Na + ) by taste buds in the oral mucosa. The high flow rate of low and medium molecular weight hyaluronic acid may enhance the perception of taste substances, while the excessive viscosity of high molecular weight hyaluronic acid may form an interfacial film, hindering the perception of taste substances by taste buds.

[0121] Example 7 Effect of hyaluronic acid concentration on sweetness perception

[0122] ① Take 1000 mL of water solution, slowly add 8 g of hyaluronic acid (or sodium hyaluronate) with a molecular weight of 100 kDa to prepare a 0.8% mass concentration of hyaluronic acid solution (or sodium hyaluronate solution). Take another 1000 mL of water, add 0.0175 g of sucralose to prepare a 87.8 μM sucralose solution, and mix the hyaluronic acid solution (or sodium hyaluronate solution) with the sucralose solution at a weight ratio of 1:1 to prepare a 43.9 μM sucralose solution containing 0.4% mass concentration of 100 kDa molecular weight of hyaluronic acid.

[0123] ② Prepare a 43.9 μM sucralose solution containing 0.4% mass concentration of 400 kDa molecular weight of hyaluronic acid in the same way.

[0124] ③ Take 1000 mL of water, slowly add 4 g of hyaluronic acid (or sodium hyaluronate solution) with a molecular weight of 100 kDa to prepare a 0.4% hyaluronic acid solution (or sodium hyaluronate solution). Take another 1000 mL of water, add 0.0175 g of sucralose to prepare a 87.8 μM sucralose solution, and mix the hyaluronic acid solution with the sucralose solution at a weight ratio of 1:1 to finally prepare a 43.9 μM sucralose solution containing 0.2% mass concentration of 100 kDa molecular weight of hyaluronic acid.

[0125] ④ Prepare a 43.9 μM sucralose solution containing 0.2% mass concentration of 400 kDa molecular weight of hyaluronic acid in the same way.

[0126] ⑤ Take 1000 mL of water, add 0.0088 g of sucralose to prepare a 43.9 μM sucralose solution.

[0127] Sensory evaluation method: 15 professional sensory evaluators taste the solutions obtained in the foregoing ①-⑤, 10 mL of the solutions obtained in the foregoing ①-⑤ are placed in 30 mL of evaluation cups and randomly presented to the sensory evaluators. The sensory evaluators spit out any solution in ①-④ after holding it in the mouth for 10 s, then rinse their mouths with clean water for 2 min, and then taste the next solution. After that, the solution ⑤ (43.9 μM sucralose solution) is compared with the solutions obtained in ①-④, and the solutions obtained in ①-④ are defined as None: no difference in sweetness; +: slightly sweeter; ++: sweeter; +++: much sweeter. Rinse your mouth for 2 min between tasting 2 samples. i.e. pure sugar water as a reference in this experiment

[0128] By Figure 18 ​The results of the experiments shown can be seen that the low molecular weight hyaluronic acid or its sodium salt in the present application (such as the hyaluronic acid with a molecular weight of 100 kDa and 400 kDa in the present embodiment) improves the sweetness perception compared to the same concentration of pure sugar water (prepared into a 43.9 μM sucralose solution). Specifically:

[0129] (1) Same concentration:

[0130] The low molecular weight hyaluronic acid has a better sweetening effect than the medium molecular weight hyaluronic acid. In the present embodiment, the 100 kDa molecular weight hyaluronic acid has a better sweetening effect than the 400 kDa molecular weight hyaluronic acid.

[0131] (2) Same molecular weight addition:

[0132] For 100 kDa molecular weight hyaluronic acid: 0.2% mass concentration of hyaluronic acid has a better sweetening effect than 0.4% mass concentration of hyaluronic acid.

[0133] For 400 kDa molecular weight hyaluronic acid: the sweetening effect at 0.2% and 0.4% mass concentration is similar, with no significant difference.

[0134] Example 8 Quantitative study of the effect of hyaluronic acid concentration on sweetness according to the sweetness-score curve

[0135] ① Sweetness-score standard curve: 4.46 g of sucralose was weighed and prepared into 5.5, 11, 22, 43.9, 87.8, 175.6, 351.2, and 702.5 μM sucralose solutions, and the scores were assigned as follows: 5.5 μM corresponds to 0.09 points, 11 μM corresponds to 0.2 points, 22 μM corresponds to 0.36 points, 43.9 μM corresponds to 0.68 points, 87.8 μM corresponds to 1.44 points, 175.6 μM corresponds to 2.36 points, 351.2 μM corresponds to 3.58 points, and 702.5 μM corresponds to 5.13 points.

[0136] ② Sweetness-score curve preparation: 4.46 g of sucralose was weighed and prepared into 5.5, 11, 22, 43.9, 87.8, 175.6, 351.2, and 702.5 μM sucralose solutions. 100 kDa and 400 kDa hyaluronic acid with a mass concentration of 0.4% were mixed with the sucrose solution at a weight ratio of 1:1. 100 kDa and 400 kDa hyaluronic acid with a mass concentration of 0.2% were prepared into 5.5, 11, 22, 43.9, 87.8, 175.6, 351.2, and 702.5 μM sucralose solutions.

[0137] Sensory evaluation method: 15 professional sensory evaluators were trained for screening. First, taste the sweetness-scoringsolution in ①, and the sweetness scores were 0.09, 0.2, 0.36, 0.68, 1.44, 2.36, 3.58, 5.13, respectively. Through 3-4 times of training, the scoring was mastered. Then, taste the solution of different sugar concentrations of hyaluronic acid with a molecular weight of 100 kDa and 400 kDa. The sample was placed in a 30 mL tasting cup, and the solution was held in the mouth for 10 s and then spit out. Scoring was performed, and the mouth was rinsed for 2 min between tasting two samples.

[0138] According to the scores of the sensory evaluators for the sugar water with different concentrations of hyaluronic acid with different molecular weights, the low-molecular-weight hyaluronic acid can enhance the sugar concentration by up to 198%, and the medium-low-molecular-weight hyaluronic acid can enhance the sugar concentration by up to 125%.

[0139] From the experimental results shown in FIGS. 1-3, it can be seen that: Figure 19

[0140] (1) The addition of medium-low-molecular-weight hyaluronic acid (such as the hyaluronic acid with a molecular weight of 100 kDa and 400 kDa in this embodiment) has different enhancement effects on the sweetness in the range of 5.5-175.6 μM sugar concentration.

[0141] (2) The low-molecular-weight hyaluronic acid has a better enhancement effect on sweetness than the medium-low-molecular-weight hyaluronic acid. Specifically:

[0142] According to the scores of the sensory evaluators for the sugar water with different concentrations of hyaluronic acid with different molecular weights, the low-molecular-weight hyaluronic acid (such as the hyaluronic acid with a molecular weight of 100 kDa in this embodiment) can increase the sugar concentration by up to 198%, and the medium-low-molecular-weight hyaluronic acid (such as the hyaluronic acid with a molecular weight of 400 kDa in this embodiment) can increase the sugar concentration by up to 125%.

[0143] Example 9: Effect of hyaluronic acid concentration on the change in sweetness intensity over time

[0144] ① 1000 mL of water solution was weighed, and 8 g of hyaluronic acid with a molecular weight of 100 kDa (or sodium hyaluronate solution) was slowly added to prepare a 0.8% hyaluronic acid solution (or sodium hyaluronate solution). In 1000 mL of water, 0.0175 g of sucralose was added to prepare a 87.8 μM sucralose solution. The hyaluronic acid solution (or sodium hyaluronate solution) and the sucralose solution were mixed in a weight ratio of 1:1. A 43.9 μM sucralose solution containing 0.4% mass concentration of 100 kDa molecular weight hyaluronic acid was prepared.

[0145] ​(2) 4.46 g sucralose was weighed and dissolved in 1000 mL water to prepare a 5.5 μM sucralose solution. 4 g of 100 kDa hyaluronic acid (or sodium hyaluronate) was added to the sucralose solution to prepare a 0.4% (mass concentration) 100 kDa hyaluronic acid (or sodium hyaluronate) solution. Another 1000 mL water was weighed and 0.0175 g sucralose was added to prepare a 87.8 μM sucralose solution. The 100 kDa hyaluronic acid (or sodium hyaluronate) solution and the sucralose solution were mixed at a weight ratio of 1:1. Finally, a 43.9 μM sucralose solution containing 0.4% (mass concentration) 100 kDa hyaluronic acid (or sodium hyaluronate) was prepared.

[0146] (3) 1000 mL water was weighed and 4 g of 100 kDa hyaluronic acid (or sodium hyaluronate) was slowly added to prepare a 0.4% (mass concentration) hyaluronic acid (or sodium hyaluronate) solution. Another 1000 mL water was weighed and 0.0175 g sucralose was added to prepare a 87.8 μM sucralose solution. The hyaluronic acid (or sodium hyaluronate) solution and the sucralose solution were mixed at a weight ratio of 1:1. Finally, a 43.9 μM sucralose solution containing 0.2% (mass concentration) 100 kDa hyaluronic acid (or sodium hyaluronate) was prepared.

[0147] (4) 4.46 g sucralose was weighed and dissolved in 1000 mL water to prepare a 5.5 μM sucralose solution. 4 g of 400 kDa hyaluronic acid was added to the sucralose solution to prepare a 0.4% (mass concentration) 400 kDa hyaluronic acid solution. Another 1000 mL water was weighed and 0.0175 g sucralose was added to prepare a 87.8 μM sucralose solution. The 400 kDa hyaluronic acid solution and the sucralose solution were mixed at a weight ratio of 1:1. Finally, a 43.9 μM sucralose solution containing 0.4% (mass concentration) 400 kDa hyaluronic acid was prepared.

[0148] (5) The sweetness-scorring curve was prepared by weighing 4.46 g sucralose to prepare 5.5, 11, 22, 43.9, 87.8, 175.6, 351.2, and 702.5 μM sucralose solutions.

[0149] Sensory evaluation method: refer to Example 3.

[0150] The results show that the addition of medium and low molecular weight hyaluronic acid can increase the intensity of sweetness, delay the time of the highest value of sweetness, and prolong the time of high sweetness intensity. Specifically, as shown in Table 1: Figure 20

[0151] (1) The addition of medium and low molecular weight hyaluronic acid (such as 100 kDa and 400 kDa hyaluronic acid in this example) can prolong the time of sweetness perception. The addition of 0.2% (mass concentration) 100 kDa hyaluronic acid can prolong the time of sweetness perception by 34.68%, and the addition of 0.4% (mass concentration) 100 kDa hyaluronic acid can prolong the time of sweetness perception by 27.78%. The addition of 0.2% (mass concentration) 400 kDa hyaluronic acid can prolong the time of sweetness perception by 40.41%, and the addition of 0.4% (mass concentration) 400 kDa hyaluronic acid can prolong the time of sweetness perception by 37.15%.

[0152] ​(2) The effect of adding medium molecular weight hyaluronic acid (such as 400 kDa hyaluronic acid in this example) on prolonging the perception of sweetness is more obvious than that of adding low molecular weight hyaluronic acid (such as 100 kDa hyaluronic acid in this example), and the highest prolongation of the perception of sweetness by adding 100 kDa hyaluronic acid is 34.68%; the highest prolongation of the perception of sweetness by adding 400 kDa hyaluronic acid is 40.41%.

[0153] (3) The effect of adding hyaluronic acid with the same molecular weight but different concentrations on the perception of sweetness is not significant.

[0154] Example 10: Effect of hyaluronic acid concentration on the perception of spiciness in a salty and spicy rice seasoning sauce

[0155] Select a certain brand of rice seasoning sauce on the market, with a salty and spicy flavor, and prepare samples A-I:

[0156] Sample A: Take 100 g of rice seasoning sauce and mix it with pure water at a weight ratio of 1:1 as the standard solution.

[0157] Sample B: Take 100 g of sample A and mix it with a solution containing 0.4% of 100 kDa hyaluronic acid at a weight ratio of 2:3, which reduces the seasoning of the sample by 60% compared to sample A;

[0158] Sample C: Take 100 g of sample A and mix it with a solution containing 0.4% of 100 kDa hyaluronic acid at a weight ratio of 4:5, which reduces the seasoning of the sample by 44.4% compared to sample A;

[0159] Sample D: Take 100 g of sample A and mix it with a solution containing 0.2% of 100 kDa hyaluronic acid at a weight ratio of 2:3, which reduces the seasoning of the sample by 60% compared to sample A;

[0160] Sample E: Take 100 g of sample A and mix it with a solution containing 0.2% of 100 kDa hyaluronic acid at a weight ratio of 4:5, which reduces the seasoning of the sample by 44.4% compared to sample A;

[0161] Sample F: Take 100 g of sample A and mix it with a solution containing 0.4% of 400 kDa hyaluronic acid at a weight ratio of 2:3, which reduces the seasoning of the sample by 60% compared to sample A;

[0162] Sample G: Take 100 g of sample A and mix it with a solution containing 0.4% of 400 kDa hyaluronic acid at a weight ratio of 4:5, which reduces the seasoning of the sample by 44.4% compared to sample A;

[0163] H sample: 100g sample A was mixed with a solution containing 0.2% 400kDa hyaluronic acid at a weight ratio of 2:3, and then the mixture was uniformly mixed. In this way, the seasoning of the sample was reduced by 60% compared with sample A;

[0164] I sample: 100g sample A was mixed with a solution containing 0.2% 400kDa hyaluronic acid at a weight ratio of 4:5, and then the mixture was uniformly mixed. In this way, the seasoning of the sample was reduced by 44.4% compared with sample A.

[0165] Fifteen sensory evaluators were selected to taste samples B and C, and 11 of them believed that the spiciness and saltiness of sample B were close to those of sample A. Therefore, it was believed that 0.4% 100kDa hyaluronic acid could reduce the content of chili sauce by 60%.

[0166] Fifteen sensory evaluators were selected to taste samples B and C, and 11 of them believed that the spiciness and saltiness of sample B were close to those of sample A. Therefore, it was believed that 0.4% 100kDa hyaluronic acid could reduce the content of chili sauce by 60%.

[0167] Fifteen sensory evaluators were selected to taste samples D and E, and 9 of them believed that the spiciness and saltiness of sample E were close to those of sample A. Therefore, it was believed that 0.2% 100kDa hyaluronic acid could reduce the content of chili sauce by 44.4%.

[0168] Fifteen sensory evaluators were selected to taste samples F and G, and 10 of them believed that the spiciness and saltiness of sample G were close to those of sample A. Therefore, it was believed that 0.4% 400kDa hyaluronic acid could reduce the content of chili sauce by 44.4%.

[0169] Fifteen sensory evaluators were selected to taste samples H and I, and 12 of them believed that the spiciness and saltiness of sample I were close to those of sample A. Therefore, it was believed that 0.2% 400kDa hyaluronic acid could reduce the content of chili sauce by 44.4%.

[0170] Therefore, for saltiness and spiciness, 100kDa hyaluronic acid has the best effect of increasing saltiness and spiciness. Among them, 0.4% 100kDa hyaluronic acid can reduce the content of chili sauce with saltiness and spiciness by 60%.

[0171] Table 1

[0172]

[0173] Example 11: Effect of Hyaluronic Acid Concentration on Perception of Spiciness in Sweet and Spicy Rice Sauce

[0174] A certain brand of rice sauce on the market was selected, which had a sweet and spicy flavor, and samples A-I were prepared.

[0175] A sample: 100g of rice sauce was mixed with pure water at a weight ratio of 1:1 as a standard solution.

[0176] B sample: 100g of sample A is weighed and mixed with a solution containing 0.4% of 100 kDa hyaluronic acid at a weight ratio of 2:3, so that the seasoning of the sample is reduced by 60% compared with sample A;

[0177] C sample: 100g of sample A is weighed and mixed with a solution containing 0.4% of 100 kDa hyaluronic acid at a weight ratio of 4:5, so that the seasoning of the sample is reduced by 44.4% compared with sample A;

[0178] D sample: 100g of sample A is weighed and mixed with a solution containing 0.2% of 100 kDa hyaluronic acid at a weight ratio of 2:3, so that the seasoning of the sample is reduced by 60% compared with sample A;

[0179] E sample: 100g of sample A is weighed and mixed with a solution containing 0.2% of 100 kDa hyaluronic acid at a weight ratio of 4:5, so that the seasoning of the sample is reduced by 44.4% compared with sample A;

[0180] F sample: 100g of sample A is weighed and mixed with a solution containing 0.4% of 400 kDa hyaluronic acid at a weight ratio of 2:3, so that the seasoning of the sample is reduced by 60% compared with sample A;

[0181] G sample: 100g of sample A is weighed and mixed with a solution containing 0.4% of 400 kDa hyaluronic acid at a weight ratio of 4:5, so that the seasoning of the sample is reduced by 44.4% compared with sample A;

[0182] H sample: 100g of sample A is weighed and mixed with a solution containing 0.2% of 400 kDa hyaluronic acid at a weight ratio of 2:3, so that the seasoning of the sample is reduced by 60% compared with sample A;

[0183] I sample: 100g of sample A is weighed and mixed with a solution containing 0.2% of 400 kDa hyaluronic acid at a weight ratio of 4:5, so that the seasoning of the sample is reduced by 44.4% compared with sample A.

[0184] Select 15 sensory evaluators to taste the above samples respectively, and compare with sample A, respectively select the samples closest to A from B-E, F-I according to their opinions.

[0185] The 15 selected sensory evaluators taste samples B and C, and 9 of them think that the spiciness and sweetness of sample B are close to those of sample A, so they think that 0.4% of 100 kDa hyaluronic acid can reduce the content of chili sauce by 60%.

[0186] Fifteen sensory panelists were selected to taste F and G samples, 12 of them thought that the spiciness and sweetness of G sample was close to that of A sample, so it was considered that 0.4% of 400 kDa hyaluronic acid could reduce the chili sauce content by 44.4%.

[0187] Fifteen sensory panelists were selected to taste F and G samples, 12 of them thought that the spiciness and sweetness of G sample was close to that of A sample, so it was considered that 0.4% of 400 kDa hyaluronic acid could reduce the chili sauce content by 44.4%.

[0188] Fifteen sensory panelists were selected to taste F and G samples, 12 of them thought that the spiciness and sweetness of G sample was close to that of A sample, so it was considered that 0.4% of 400 kDa hyaluronic acid could reduce the chili sauce content by 44.4%.

[0189] Therefore, it is considered that 100 kDa hyaluronic acid has the best sweetening and spicing effect for sweet and spicy taste; among them, the addition amount of 0.2% of 100 kDa hyaluronic acid can reduce the chili sauce content of sweet and spicy taste by 60%.

[0190] Table 2

[0191]

[0192] Example 12 to explore the effect of hyaluronic acid concentration on the perception of spiciness with pickled pepper flavored rice sauce

[0193] Select a certain brand of rice sauce on the market, the taste is pickled pepper flavor, sample A 100g chili sauce is mixed with pure water solution in a weight ratio of 1:1 as a standard solution.

[0194] Sample B 100g chili sauce is mixed with 0.4% 100 kDa hyaluronic acid solution in a weight ratio of 2:3, and the seasoning of the sample is reduced by 60% compared with sample A;

[0195] Sample C 100g sample A is mixed with 0.4% 100 kDa hyaluronic acid solution in a weight ratio of 4:5, and the seasoning of the sample is reduced by 44.4% compared with sample A;

[0196] Sample D 100g sample A is mixed with 0.2% 100 kDa hyaluronic acid solution in a weight ratio of 2:3, and the seasoning of the sample is reduced by 60% compared with sample A;

[0197] Sample E 100g sample A is mixed with 0.2% 100 kDa hyaluronic acid solution in a weight ratio of 4:5, and the seasoning of the sample is reduced by 44.4% compared with sample A;

[0198] F sample 100g sample A is weighed, mixed with 0.4% 400kDa hyaluronic acid solution at a weight ratio of 2:3, and then mixed uniformly. The seasoning of this sample is reduced by 60% compared with sample A.

[0199] G sample 100g sample A is weighed, mixed with 0.4% 400kDa hyaluronic acid solution at a weight ratio of 4:5, and then mixed uniformly. The seasoning of this sample is reduced by 44.4% compared with sample A.

[0200] H sample 100g sample A is weighed, mixed with 0.2% 400kDa hyaluronic acid solution at a weight ratio of 2:3, and then mixed uniformly. The seasoning of this sample is reduced by 60% compared with sample A.

[0201] I sample 100g sample A is weighed, mixed with 0.2% 400kDa hyaluronic acid solution at a weight ratio of 4:5, and then mixed uniformly. The seasoning of this sample is reduced by 44.4% compared with sample A.

[0202] Select 15 sensory evaluators to taste the above samples respectively, and compare them with sample A. Select the samples from B-E, F-I that they think have the closest acidity and spiciness to A.

[0203] Of the 15 selected sensory evaluators, 11 think that sample B has a spiciness and acidity close to that of sample A after tasting samples B and C. Therefore, it is believed that 0.4% 100kDa hyaluronic acid can reduce the content of chili sauce by 60%.

[0204] Of the 15 selected sensory evaluators, 9 think that sample D has a spiciness and acidity close to that of sample A after tasting samples D and E. Therefore, it is believed that 0.2% 100kDa hyaluronic acid can reduce the content of chili sauce by 60%.

[0205] Of the 15 selected sensory evaluators, 10 think that sample G has a spiciness and acidity close to that of sample A after tasting samples F and G. Therefore, it is believed that 0.4% 400kDa hyaluronic acid can reduce the content of chili sauce by 44.4%.

[0206] Of the 15 selected sensory evaluators, 11 think that sample I has a spiciness and acidity close to that of sample A after tasting samples H and I. Therefore, it is believed that 0.2% 400kDa hyaluronic acid can reduce the content of chili sauce by 44.4%.

[0207] Therefore, it is believed that 100kDa hyaluronic acid has the best effect on increasing spiciness for pickled pepper flavor. The addition of 0.4% 100kDa hyaluronic acid can reduce the content of chili sauce for pickled pepper flavor by 60%.

[0208] Table 3

[0209]

[0210] Example 13 investigates the effect of hyaluronic acid concentration on bitterness perception with a bitter coffee

[0211] Select a brand of coffee freeze-dried pure American instant coffee without sugar, select coffee powder No. 117, which has a stronger bitter taste.

[0212] A sample of 14g of coffee powder was added to 1000mL of 90°C hot water, mixed evenly;

[0213] The sample B was 14g of coffee powder added to 1500mL of 90°C 0.4% concentration of 100kDa hyaluronic acid solution, then the bitter taste of the sample was reduced by 33.3% compared with the bitter taste of the sample A;

[0214] The sample C was 14g of coffee powder added to 1250mL of 90°C 0.4% concentration of 100kDa hyaluronic acid solution, then the bitter taste of the sample was reduced by 20% compared with the bitter taste of the sample A;

[0215] The sample D was 14g of coffee powder added to 1500mL of 90°C 0.2% concentration of 100kDa hyaluronic acid solution, then the bitter taste of the sample was reduced by 33.3% compared with the bitter taste of the sample A;

[0216] The sample E was 14g of coffee powder added to 1250mL of 90°C 0.2% concentration of 100kDa hyaluronic acid solution, then the bitter taste of the sample was reduced by 20% compared with the bitter taste of the sample A;

[0217] The sample F was 14g of coffee powder added to 1500mL of 90°C 0.4% concentration of 400kDa hyaluronic acid solution, then the bitter taste of the sample was reduced by 33.3% compared with the bitter taste of the sample A;

[0218] The sample G was 14g of coffee powder added to 1250mL of 90°C 0.4% concentration of 400kDa hyaluronic acid solution, then the bitter taste of the sample was reduced by 20% compared with the bitter taste of the sample A;

[0219] The sample H was 14g of coffee powder added to 1500mL of 90°C 0.2% concentration of 400kDa hyaluronic acid solution, then the bitter taste of the sample was reduced by 33.3% compared with the bitter taste of the sample A;

[0220] The sample I was 14g of coffee powder added to 1250mL of 90°C 0.2% concentration of 400kDa hyaluronic acid solution, then the bitter taste of the sample was reduced by 20% compared with the bitter taste of the sample A.

[0221] Select 15 sensory evaluators to taste the above samples respectively, compare with sample A, and select the samples from B-E, F-I that they think have the closest bitterness to A.

[0222] 15 panelists tasted B and C samples, 8 panelists thought that the bitterness of B sample was close to A sample, so 0.4% 100 kDa HA could reduce 33.3% coffee content.

[0223] 15 panelists tasted D and E samples, 10 panelists thought that the bitterness of D sample was close to A sample, so 0.2% 100 kDa HA could reduce 33.3% coffee content.

[0224] 15 panelists tasted F and G samples, 9 panelists thought that the bitterness of G sample was close to A sample, so 0.4% 400 kDa HA could reduce 20% coffee content.

[0225] 15 panelists tasted H and I samples, 12 panelists thought that the bitterness and sourness of I sample was close to A sample, so 0.2% 400 kDa HA could reduce 20% coffee content.

[0226] So for bitter coffee, 100 kDa HA was the best to enhance the bitterness, and 0.2% 100 kDa HA could reduce 33.3% coffee content.

[0227] Table 4

[0228]

[0229] Example 14: Effect of HA concentration on sourness perception for sour coffee

[0230] Select a brand of coffee freeze-dried pure American instant coffee without sugar, select coffee powder No. 114 with stronger sourness.

[0231] A sample: 14 g of coffee powder was added to 1000 mL of 90°C hot water and mixed evenly;

[0232] B sample: 14 g of coffee powder was added to 1500 mL of 90°C 0.4% 100 kDa HA solution, so the bitter taste compounds of the sample were reduced by 33.3% compared with A sample;

[0233] C sample: 14 g of coffee powder was added to 1250 mL of 90°C 0.4% 100 kDa HA solution, so the bitter taste compounds of the sample were reduced by 20% compared with A sample;

[0234] D sample 14g coffee powder was added to 1500ml of 90°C 0.2% concentration of 100kDa hyaluronic acid solution, the bitter taste of the sample was reduced by 33.3% than sample A;

[0235] E sample 14g coffee powder was added to 1250ml of 90°C 0.2% concentration of 100kDa hyaluronic acid solution, the bitter taste of the sample was reduced by 20% than sample A;

[0236] F sample 14g coffee powder was added to 1500ml of 90°C 0.4% concentration of 400kDa hyaluronic acid solution, the bitter taste of the sample was reduced by 33.3% than sample A;

[0237] G sample 14g coffee powder was added to 1250ml of 90°C 0.4% concentration of 400kDa hyaluronic acid solution, the bitter taste of the sample was reduced by 20% than sample A;

[0238] H sample 14g coffee powder was added to 1500ml of 90°C 0.2% concentration of 400kDa hyaluronic acid solution, the bitter taste of the sample was reduced by 33.3% than sample A;

[0239] I sample 14g coffee powder was added to 1250ml of 90°C 0.2% concentration of 400kDa hyaluronic acid solution, the bitter taste of the sample was reduced by 20% than sample A.

[0240] 15 sensory evaluation staff were selected to taste the above samples, and compared with sample A, they selected the samples from B-E, F-I which they thought the acidity was closest to sample A.

[0241] 15 sensory evaluation staff were selected to taste samples B and C, 10 of them thought that sample C was closest to sample A in acidity, so they thought that 0.4% of 100kDa hyaluronic acid could reduce the coffee content by 20%.

[0242] 15 sensory evaluation staff were selected to taste samples D and E, 12 of them thought that sample D was closest to sample A in acidity, so they thought that 0.2% of 100kDa hyaluronic acid could reduce the coffee content by 33.3%.

[0243] 15 sensory evaluation staff were selected to taste samples F and G, 11 of them thought that sample G was closest to sample A in acidity, so they thought that 0.4% of 400kDa hyaluronic acid could reduce the coffee content by 20%.

[0244] The 15 selected sensory panelists tasted the H and I samples, and 9 of them thought that the acidity of the H sample was close to that of the A sample, so it was considered that 0.2% of 400 kDa hyaluronic acid could reduce the coffee content by 33.3%.

[0245] Therefore, it is considered that 100 kDa hyaluronic acid is optimal for enhancing the sour taste effect for sour coffee; and 0.2% of 100 kDa hyaluronic acid can reduce the coffee content by 33.3%.

[0246] Table 5

[0247]

[0248]

[0249] Example 15 Study on the mechanism of hyaluronic acid affecting taste perception by isothermal titration calorimetry (ITC)

[0250] Weigh 20 mL of PBS buffer solution and add 20 mg of Type III porcine stomach-derived mucin, stir overnight at 12°C to fully hydrate. Centrifuge at 800g for 10 min to remove insoluble precipitates.

[0251] First, the mucin solution and the hyaluronic acid solution were fully dialyzed in PBS buffer solution at pH 6.7 for 24 h. The dialyzed PBS buffer solution was used as the dilution solution, and water and the blank control group were used as water. After dialysis, the concentration of mucin was determined by the BCA method.

[0252] The ITC experiment was performed on a Micro VP ITC200 microcalorimeter. The low, medium and high (8 kDa, 100 kDa and 400 kDa) molecular weight hyaluronic acid solution was sucked into a 300 μL titration needle, and the air bubbles were removed. Mucin was added to a 1400 μL sample cell. The whole titration process was carried out at 25°C, a total of 28 times, each time 10 μL, the stirring rate was 300 rpm, and the time interval between each injection was 300 s. The experimental data were further fitted using MicroCal Origin ITC analysis software.

[0253] First, the thermodynamic changes of hyaluronic acid and mucin of different molecular weights when they interacted with each other were compared. The interaction between hyaluronic acid and mucin was exothermic under the experimental conditions, and the interaction between the two was enhanced with the increase of the molecular weight of hyaluronic acid. These interactions will directly affect the taste buds in the oral mucosa to taste substances (such as Na +The effect of the generation or enhancement or inhibition is related to the molecular weight of the hyaluronic acid and the molecular conformation in the solution. For example, the low and medium molecular weight with greater flexibility and stronger activity of the molecular chain can promote the contact between the taste substance and the taste bud in the oral mucosa, thereby achieving the effect of taste enhancement. However, if the molecular weight is too large, the diffusion of the taste substance (such as Na + ) in the mucosa layer is hindered, and the perception of the taste substance (such as Na + ) by the taste bud in the oral mucosa is inhibited.

Claims

1. Use of hyaluronic acid as a single component in taste enhancement, characterized in that, The taste enhancement refers to enhancing the taste perception of taste buds of the human body, so that the human body reduces the intake of spicy taste substances in food in the case of obtaining the same taste perception; the food includes condiments; The hyaluronic acid refers to a natural high-molecular linear polysaccharide formed by repeating connection of disaccharide units composed of D-glucuronic acid and N-acetylglucosamine, the molecular weight of the hyaluronic acid is 100 kDa, and the amount is 0.2% or 0.4% of the mass of the food.

2. Use according to claim 1, characterized in that, The condiments include solid condiments, semi-solid condiments and liquid condiments.

3. Use according to claim 2, characterized in that, The condiments are sauces, including soybean sauce and bean paste.

4. Use according to claim 1, characterized in that, The food includes meat products, instant food, potato food, pickled vegetables and reconstituted eggs.

5. Use according to claim 1, characterized in that, The condiments made of hyaluronic acid are added in the process of making or eating food.

6. Use of hyaluronic acid as a single component in taste enhancement, characterized in that, The taste enhancement refers to enhancing the taste perception of taste buds of the human body, so that the human body reduces the intake of bitter taste substances in coffee in the case of obtaining the same taste perception; The hyaluronic acid refers to a natural high-molecular linear polysaccharide formed by repeating connection of disaccharide units composed of D-glucuronic acid and N-acetylglucosamine, the molecular weight of the hyaluronic acid is 100 kDa, and the amount is 0.2% or 0.4% of the mass of the coffee.

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