Use of allulose disaccharides for inhibiting hydroxymethylfurfural (HMF) formation

Abstract

To provide, in heating a food including saccharide, a composition for inhibiting generation of HMF (hydroxymethylfurfural) and / or for preventing browning, and methods for inhibiting generation of HMF and preventing browning.SOLUTION: A composition for inhibiting generation of HMF and / or for preventing browning includes allulose disaccharide. In the allulose disaccharide, two allulose molecules are coupled with each other by glycosidic bond. In the glycosidic bond, a hydroxy group of C2 of one allulose molecule of the two allulose molecules is coupled with any one of C1-C6 of another allulose molecule by glycosidic bond.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present application relates to novel uses of allulose disaccharides. [Background technology]

[0002] HMF is an organic compound formed by the dehydration of compounds containing aldehyde and hydroxyl groups. It is a white, low-melting compound that is highly soluble in both water and organic solvents. High levels of HMF production suggest a high likelihood of product freshness loss during processing and distribution, quality deterioration due to accumulated heat damage, and even persistent non-enzymatic browning. Furthermore, HMF poses unresolved negative concerns regarding human health risks, such as the persistence of carcinogenicity (Abraham K (2011) Toxicology and risk assessment of 5-hydroxymethylfurfural in food, Molecular Nutrition & Food Research. 55 (5): 667-678 (Non-Patent Document 1)).

[0003] Heat sterilization is the most commonly used method in food processing, but as mentioned above, foods containing sugars generate HMF when heated, so reducing the amount of HMF produced is an important issue.

[0004] To solve this problem, research has been conducted into methods for stabilizing product quality by suppressing HMF production, as reported in, for example, Food Browning and Its Prevention: An Overview. J. Agric. Food Chem., Vol. 44, No. 3 (Non-Patent Document 2). However, research into more effective methods is still needed, and no methods involving the addition of carbohydrates have yet been reported. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] US Patent Application Publication No. 2018 / 0327796 [Non-patent literature]

[0006] [Non-Patent Document 1] Abraham K (2011) Toxicology and risk assessment of 5-Hydroxymethylfurfural in food, Molecular Nutrition & Food Research. 55 (5): 667-678 [Non-patent document 2] Food Browning and Its Prevention: An Overview. J. Agric. Food Chem., Vol. 44, No. 3 Summary of the Invention [Problem to be solved by the invention]

[0007] Against this background, the present inventors isolated a novel substance, confirmed that this substance was allulose disaccharide, and confirmed that when allulose disaccharide is added to a composition containing sugars, the production of HMF is suppressed, thereby completing the present invention. [Means for solving the problem]

[0008] The present application provides a composition for inhibiting HMF (hydroxymethylfurfural) production and / or preventing browning, which comprises an allulose disaccharide.

[0009] The present application provides a method for inhibiting HMF production and / or preventing browning, comprising providing a composition comprising a sugar and an allulose disaccharide.

[0010] The present application provides a method for producing a composition containing a sugar, the method comprising: preparing a mixed composition containing a sugar and an allulose disaccharide; and heating the mixed composition. [Effects of the Invention]

[0011] The composition containing the allulose disaccharide of the present application inhibits the production of harmful substances that are generated during processing, sterilization, and long-term storage, and can therefore be useful for inhibiting sugar dehydration, inhibiting HMF production, and / or preventing browning. [Brief explanation of the drawings]

[0012] [Figure 1] 1 is an HPLC chromatogram of disaccharides produced during the allulose production process analyzed on a column (Biorad Aminex HPX-87C). [Figure 2] HPLC chromatograms of D1 and D2 obtained by analyzing a mixture of disaccharides produced during the allulose production process using a column (YMC Pack Polyamine II). [Figure 3] The three-dimensional structure of D1, an allulose disaccharide, is shown. [Figure 4] The structure and carbon numbering of allulose are shown. DETAILED DESCRIPTION OF THE INVENTION

[0013] This will be explained in more detail as follows. Meanwhile, each description and embodiment disclosed in this application can also be applied to each other description and embodiment. In other words, any combination of various elements disclosed in this application belongs to the category of this application. Furthermore, the specific description described below is not considered to limit the category of this application.

[0014] Additionally, those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the present application described herein which equivalents are intended to be encompassed by this application.

[0015] One aspect of the present application provides a composition for inhibiting HMF (hydroxymethylfurfural) production, which contains allulose disaccharide.

[0016] In the present application, the term "allulose disaccharide" can be used interchangeably with terms such as "allulose dimer," "allulose diploid," and "disaccharide allulose," and means "a compound in which two allulose molecules are linked by a glycosidic bond."

[0017] Specifically, the allulose disaccharide may be formed by linking two allulose molecules by a glycosidic bond, in which the hydroxy group at the carbon 2 (C2) of one of the two allulose molecules is glycosidic bonded to the hydroxy group at any one of the carbons 1 to 6 (C1 to C6) of the other allulose molecule.

[0018] Specifically, at least one of the two allulose molecules may be a cyclic allulose, and the hydroxyl group at the carbon 2 of the cyclic allulose may be linked to the hydroxyl group at any one of the carbons 1 to 6 of the other allulose molecule via a glycosidic bond. The number of glycosidic bonds may be one or two, and specifically may be one.

[0019] In one embodiment, the bond may be a glycosidic bond between the hydroxy group at the 2-carbon of a cyclic allulose and the hydroxy group at the 6-carbon of another allulose.

[0020] In one embodiment, one of the two allulose molecules may be in the form of psicofuranose, and the other may be in the form of psicopyranose. In one embodiment, the allulose may be represented by, but is not limited to, the following chemical formula (1):

[0021] [ka] ···(1)

[0022] In one embodiment, the allulose disaccharide of the present application may be a compound named 2-(hydroxymethyl)-2-((3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol, more specifically, (2S,3R,4R,5R)-2-(hydroxymethyl)tetrahydrofuran-2-yl The compound may be, but is not limited to, a compound named ((2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol.

[0023] In one embodiment, the allulose disaccharide of the present application may be a compound named 2-(hydroxymethyl)-2-((3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol, more specifically, (2S,3R,4R,5R)-2-(hydroxymethyl)tetrahydrofuran-2-yl The compound may be, but is not limited to, a compound named ((2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol.

[0024] The (2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol can collectively refer to compounds named 6-O-β-D-Psicopyranosyl-α-D-psicofuranose or 6-O-β-D-Psicopyranosyl-β-D-psicofuranose, depending on the form of the psicofuranose.

[0025] The (2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol is (2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R,5S)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol The compound may be, but is not limited to, a compound designated as (2S,3R,4R,5R)-2-(hydroxymethyl)-2-(((2R,3S,4R,5R)-3,4,5-trihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)methoxy)tetrahydro-2H-pyran-3,4,5-triol.

[0026] Specifically, the compound of the formula (1) exists in two forms of the following formulas (2) and / or (3).

[0027] [ka] ···(2)

[0028] [ka] ···(3)

[0029] The compound of chemical formula (2) is 6-O-β-D-psicopyranosyl-α-D-psico furanose, and the compound of chemical formula (3) can be called 6-O-β-D-psicopyranosyl-β-D-psico furanose.

[0030] "HMF" in the present application is a substance also known as 5'-HMF (5-hydroxymethyl-furfural), and may have a structure represented by the following chemical formula (4).

[0031] [ka] ···(4)

[0032] The HMF may be produced from a compound containing an aldehyde group and a hydroxy group.

[0033] In this application, "HMF-producing substance" means a compound that produces HMF.

[0034] The HMF-producing substances are not limited as long as they contain an aldehyde group and / or a hydroxy group, and may include, for example, carbohydrates, glycolipids, glycoproteins, and other substances.

[0035] Specifically, the HMF may be produced by sugars (represented by sugar or saccharide). The HMF may be produced by glycolysis, which includes glycolysis by dehydration.

[0036] In particular, the sugar may be a monosaccharide. The monosaccharides include, without limitation, aldotriose (glyceraldehyde), ketotriose (dihydroxyacetone), aldotetrose (erythrose, threose), ketotetrose (erythrulose), aldopentose (arabinose, lyxose, ribose, xylose (wood sugar, wood sugar)), ketopentose (ribulose, xylulose), deoxysugar (deoxyribose), aldohexose (allose, altrose, galactose, glucose (grape sugar)), gulose, idose, mannose, talose), ketohexose (fructose (fructose), allulose, sorbose, tagatose), deoxysugar (fucose, fuculose, rhamnose), ketoheptose (mannoheptulose, sedoheptulose), and the like, as long as they have the potential to produce HMF. Specifically, it may be a hexose, more specifically, it may be allulose, but is not limited to these.

[0037] Alternatively, HMF-producing materials may also include mixtures containing one or more of the aforementioned materials.

[0038] HMF is also an example of a glycation intermediate. The term "glycation product" as used herein refers to a product formed by a non-enzymatic reaction between an amino acid group, such as a lysine residue in a protein, and a reducing sugar. It encompasses both glycation intermediates and advanced glycation end products, and is formed as advanced glycation end products (AGEs) from glycation intermediates. Advanced glycation end products may be brown in color and produce volatile aroma compounds, or may refer to a variety of substances formed in the body by the reaction of blood glucose or glucose degradation products with various protein components, such as hemoglobin, LDL, and collagen. Glycation products are representative examples of by-products formed during the processing, storage, and sterilization of sugar-containing compositions.

[0039] Once generated, advanced glycation end products (ADGs) are not broken down even when blood glucose levels return to normal, but instead accumulate in the blood and tissues during the life of the protein. Accumulated ADGs form cross-links with proteins and interact with the receptor for advanced glycation end products (RAGE), leading to the accumulation of inflammatory cells.

[0040] Therefore, the production of advanced glycation end products, which can have harmful effects on the human body, is closely related to the amount of HMF, and the allulose disaccharide of the present application may also inhibit the production of advanced glycation end products by inhibiting the production of HMF.

[0041] That is, another aspect of the present application provides a composition for inhibiting the production of glycation products, which comprises allulose disaccharide.

[0042] Another aspect of the present application provides a composition for inhibiting sugar dehydration, comprising an allulose disaccharide.

[0043] The term "dehydration" in the present application refers to any process in which water is separated intramolecularly or intermolecularly. In the present application, the molecule causing the dehydration may be a sugar molecule.

[0044] In the present application, the term "sugar dehydration reaction" refers to a reaction in which HO is produced within a sugar molecule or between sugar molecules. Specifically, the sugar dehydration reaction may be a reaction in which HO is produced within a sugar molecule.

[0045] Specifically, the sugar molecule may be a monosaccharide, which is a unit of sugar or glucide. The monosaccharide is as described above.

[0046] When a dehydration reaction occurs in a sugar molecule, other substances derived from the sugar molecule may be produced in addition to the molecule HO. For example, the dehydration reaction in a hexose may produce hydroxymethylfurfural (HMF) in addition to the molecule HO, as shown in Reaction Scheme 1 below.

[0047] Reaction Scheme 1 [ka]

[0048] "Inhibition of sugar dehydration" means preventing the aforementioned sugar dehydration reaction or reducing the sugar dehydration reaction compared to an environment in which allulose disaccharide is absent or present in relatively small amounts. Such inhibition of sugar dehydration can be confirmed by measuring the amount of sugar dehydration products. For example, the amount of HMF produced can be measured to confirm whether sugar dehydration is inhibited.

[0049] The dehydration reaction may be carried out under heating, sterilization, and / or known processing conditions for the composition, but is not limited thereto, and includes reactions that occur naturally at room temperature, and therefore reactions that occur during storage of the composition.

[0050] Another aspect of the present application provides a browning prevention composition comprising an allulose disaccharide.

[0051] In this application, the term "anti-browning" may be used to mean preventing browning, delaying browning, inhibiting browning, etc., and these terms may be used interchangeably in this application. For example, browning in foods such as cereals and cereal bars, potato chips, bread, carbonated drinks, fruit and vegetable juices, fruit juices, fruit wines, sauces, candies, jellies, jams, ice cream, and beer causes deterioration in quality leading to loss of aroma, taste, and nutritional value.

[0052] Browning reactions include the Maillard reaction and caramelization. For example, the Maillard reaction occurs when the carbonyl groups of sugars react with the amino acid groups of proteins under heat to produce brown substances (melanoidins). This reaction is also called the melanoidin reaction, or the aminocarbonyl reaction caused by the reactants.

[0053] In the intermediate stages of the Maillard reaction, a highly reactive substance called HMF is produced, and the product in the final stage of the reaction is also a highly reactive substance. These substances form polymers to form melanoidin pigments, which have brown fluorescent properties, and this process causes browning.

[0054] As mentioned above, the production of HMF is closely related to the browning reaction of the composition, so the allulose disaccharide of the present application can be used for preventing browning by inhibiting the production of HMF, i.e., the prevention of browning can be due to the inhibition of HMF production.

[0055] The content of allulose disaccharide in the composition may be more than 0 and not more than 15 parts by weight per 100 parts by weight of the HMF-producing substance contained in the composition.

[0056] Specifically, the allulose disaccharide may be contained in an amount of more than 0.0001 parts by weight, more than 0.001 parts by weight, more than 0.01 parts by weight, more than 0.1 parts by weight, or more than 0.15 parts by weight but not more than 15 parts by weight per 100 parts by weight of the total HMF-producing material, and / or may be contained in an amount of less than 15 parts by weight, less than 13 parts by weight, less than 11 parts by weight, less than 10 parts by weight, less than 9 parts by weight, less than 8 parts by weight, less than 7 parts by weight, less than 6 parts by weight, less than 5 parts by weight, less than 4 parts by weight, less than 3 parts by weight, less than 2 parts by weight, or less than 1 part by weight but more than 0 parts by weight per 100 parts by weight of the HMF-producing material, but is not limited to these.

[0057] The content of allulose disaccharide in the composition may be more than 0 and not more than 15 parts by weight per 100 parts by weight of the total sugars contained in the composition. Specifically, the content may be more than 0.0001 parts by weight, more than 0.001 parts by weight, more than 0.01 parts by weight, more than 0.1 parts by weight, or more than 0.15 parts by weight but not more than 15 parts by weight per 100 parts by weight of the total sugars, and / or the content may be 15 parts by weight or less, 13 parts by weight or less, 11 parts by weight or less, 10 parts by weight or less, 9 parts by weight or less, 8 parts by weight or less, 7 parts by weight or less, 6 parts by weight or less, 5 parts by weight or less, 4 parts by weight or less, 3 parts by weight or less, or 2 parts by weight or less but more than 0 parts by weight per 100 parts by weight of the total sugars, but is not limited thereto.

[0058] Alternatively, the content of allulose disaccharide in the composition may be more than 0 and not more than 15 parts by weight per 100 parts by weight of the total solid content of the composition. Specifically, the content may be more than 0.0001 parts by weight, more than 0.001 parts by weight, more than 0.01 parts by weight, more than 0.1 parts by weight, or more than 0.15 parts by weight but not more than 15 parts by weight per 100 parts by weight of the total solid content, or may be 15 parts by weight or less, 13 parts by weight or less, 11 parts by weight or less, 10 parts by weight or less, 9 parts by weight or less, 8 parts by weight or less, 7 parts by weight or less, 6 parts by weight or less, 5 parts by weight or less, 4 parts by weight or less, 3 parts by weight or less, or 2 parts by weight or less but more than 0 part by weight per 100 parts by weight of the total solid content, but is not limited thereto.

[0059] The composition may be a food composition. Food compositions of the present application include, but are not limited to, general foods, health foods, and medical (or patient) food compositions. Specifically, the food compositions of the present application may be beverages (e.g., carbonated beverages, fruit juice drinks, fruit and vegetable drinks, dietary fiber drinks, carbonated water, miso soup, tea, coffee, etc.), alcoholic beverages, bread, sauces (e.g., ketchup, pork cutlet sauce, etc.), dairy products (e.g., fermented milk, processed milk, etc.), meat products (e.g., ham, sausage, jerky, etc.), chocolate products, gum, candy, jelly, ice cream, syrup, dressing, snacks (e.g., cookies, crackers, biscuits, etc.), pickled fruits and vegetables (e.g., candied fruits, pickled fruits, red ginseng extract, red ginseng slices, etc.), meal replacement foods (e.g., frozen foods, retort foods, HMR (home meal replacement), etc.), or processed foods. However, these are merely examples and are not intended to be limiting.

[0060] The food compositions of the present application may contain various flavoring agents and natural carbohydrates as additional ingredients. The natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of sweeteners that can be used include natural sweeteners such as thaumatin and stevia extract, and synthetic sweeteners such as sucralose, saccharin, and aspartame.

[0061] In addition to the above, the food composition of the present application may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectin and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, acidity and salinity agents used in carbonated beverages, etc. The food composition of the present application may also contain fruit pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients may be used independently or in combination. Furthermore, those skilled in the art may appropriately select and add substances typically contained in food compositions. The proportion of these additives may be selected from the range of 0.001 to 1 part by weight or 0.01 to 0.20 parts by weight per 100 parts by weight of the food composition of the present application, but is not limited thereto.

[0062] Another aspect of the present application provides a method for inhibiting HMF production, comprising preparing a composition comprising a sugar and an allulose disaccharide.

[0063] In this application, "prepare" includes, without limitation, any method of providing a composition containing a sugar and an allulose disaccharide. That is, it includes any method of causing a composition to contain a sugar and an allulose disaccharide. For example, preparing a composition containing a sugar and an allulose disaccharide can include adding allulose disaccharide to a composition containing a sugar, adding a sugar to a composition containing allulose disaccharide, or generating allulose disaccharide during the production of a sugar / sugar composition.

[0064] The composition may contain other ingredients without limitation as long as it contains sugars and allulose disaccharides.

[0065] Meanwhile, a composition containing sugars and allulose disaccharides is also called a "mixture." In the mixture composition, the "sugars" other than allulose disaccharides may include, but are not limited to, allulose.

[0066] The method for inhibiting HMF production may further include heating the composition after preparing the composition containing the saccharide and allulose disaccharide, but is not limited thereto, and the heating may be performed before, after, or simultaneously with the preparation of the mixed composition.

[0067] The heating may be performed within an appropriate temperature range depending on the type of composition, and the temperature range, heating time, sterilization method, etc. can be appropriately determined by a person skilled in the art based on what is known in the art. Specifically, the heating may be performed at a temperature of 60°C to 100°C, more specifically, at a temperature of 60°C to 95°C, 65°C to 95°C, or 70°C to 95°C, but is not limited thereto.

[0068] The heating may be carried out for a time period of more than 0 hours and not more than 108 hours, specifically, for 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours or more, but is not limited to these.

[0069] Another aspect of the present application provides a method for producing a composition containing a sugar, the method comprising: preparing a mixed composition containing a sugar and an allulose disaccharide; and heating the mixed composition.

[0070] Heating the mixed composition may be, but is not limited to, inhibiting HMF production.

[0071] The heating can be carried out before, after or simultaneously with the preparation of the mixed composition.

[0072] The sugar-containing composition produced by the above production method may have a low impurity content, a low HMF production amount, an increased allulose content, a change in its physical properties, a low production of by-products, or a low occurrence of crystallization, browning reactions, oxidation / reduction reactions, or reactions in which sugars other than allulose disaccharides are converted into other substances. Specifically, the sugar-containing composition may have a low occurrence of the above-mentioned reactions compared to when a composition containing no allulose disaccharides or a composition containing a relatively low amount of allulose disaccharides compared to the above-mentioned mixed composition is heated under the same conditions. However, the sugar-containing composition is not limited to these.

[0073] Another aspect of the present application provides a method for inhibiting sugar dehydration, comprising providing a composition comprising a sugar and an allulose disaccharide.

[0074] Another aspect of the present application provides a method for preventing browning, comprising providing a composition comprising a sugar and an allulose disaccharide.

[0075] Another aspect of the present application provides a method for sterilizing a composition, comprising providing a composition comprising a sugar and an allulose disaccharide.

[0076] The method may further include heating the composition after preparing the composition containing the saccharide and allulose disaccharide, but is not limited thereto, and the heating may be performed before, after, or simultaneously with the preparation of the mixed composition.

[0077] HMF production, sugar dehydration reaction, prevention of browning, sugars, and heating are as described above.

[0078] The composition may be, by way of example only, a food composition, but is not limited thereto.

[0079] The food items have been mentioned above.

[0080] Another aspect of the present application provides a composition for inhibiting saccharide denaturation, comprising allulose disaccharide.

[0081] Another aspect of the present application provides a method for inhibiting denaturation of a composition, the method comprising providing a composition comprising a sugar and an allulose disaccharide.

[0082] The method may further include heating the composition after preparing the composition containing the saccharide and allulose disaccharide, but is not limited thereto, and the heating may be performed before, after, or simultaneously with the preparation of the mixed composition.

[0083] The allulose disaccharide, the sugar, and the heating are as described above.

[0084] The denaturation includes, but is not limited to, crystallization, browning, oxidation / reduction, etc., in which sugars are converted into other substances, their physical properties are changed, or by-products are generated.

[0085] Another aspect of the present application provides the use of allulose disaccharides for inhibiting HMF production.

[0086] The allulose disaccharide and the inhibition of HMF production are as described above. [Example]

[0087] The present application will be described in more detail below through examples and experimental examples. However, these examples and experimental examples are intended to exemplify the present application and are not intended to limit the scope of the present application.

[0088] Example 1: Isolation of novel allulose disaccharides In the process of producing allulose, which is disclosed in US 2018-0327796 A1 (Patent Document 1), disaccharides were separated by HPLC. Specifically, the HPLC chromatogram analysis conditions shown in Table 1 below were followed, and it was confirmed that a previously unknown novel substance was produced from the raw solution in addition to allulose, as shown in Figure 1.

[0089] The content of the novel substance isolated as described above varied slightly depending on the manufacturing process, but it was confirmed that it was present at less than 2% in the initial raw solution and increased to a level of 5% depending on the storage time.

[0090] [Table 1]

[0091] As a result, allulose was confirmed at 21.1 minutes and the new substance at 31.7 minutes.

[0092] In order to separate the resulting novel substance, it was purified to a purity of 95% or more using preparative HPLC, and then precisely separated again using a normal phase column.

[0093] Specifically, HPLC chromatograms were performed.

[0094] The chromatographic separation conditions are as shown in Table 2 below.

[0095] [Table 2]

[0096] As a result, it was confirmed that a substance that showed one peak under the HPLC conditions in Table 1 showed two peaks under the separation conditions in Table 2, and they were separated (Figure 2). The substance with the peak identified at 22.5 minutes was named D1, and the substance with the peak identified at 17.7 minutes was named D2.

[0097] Example 2: Confirmation of the HMF production inhibitory effect of allulose disaccharide Example 2-1: Comparison of HMF production rates of allulose disaccharides Allulose was selected as a representative example of a monosaccharide that denatures upon heating and produces a high amount of HMF. To determine whether the disaccharides isolated in Example 1 could be applied to various types of foods containing different amounts of sugar, the differences were compared by varying the sugar concentration. Furthermore, considering a more severe environment, the HMF production rate was compared under the retort environment (121°C, 15 minutes), which is the highest temperature among food sterilization conditions.

[0098] Specifically, crystalline allulose (CJ CheilJedang, purity 99% or higher), which has the highest composition ratio of any monosaccharide, was added to ultrapure water without impurities, and the disaccharides with a purity of 95% or higher separated in Example 1 were quantitatively measured to produce Experimental Example 1 at different mixing ratios. Furthermore, samples (A) to (F) were prepared by adding different amounts of water to Experimental Example 1 to achieve concentrations of 1, 5, 10, 20, 30, and 50% (w / w) (Table 3).

[0099] Meanwhile, to compare the effects of other disaccharides, sugar, a typical disaccharide, was added instead of allulose disaccharide to prepare Comparative Example 1. Specifically, sugar, a disaccharide, was added to allulose crystals, a monosaccharide, in the same ratio as in Experimental Example 1 using the same method as in Experimental Example 1, and the proportions of the components were confirmed using HPLC under the conditions in Table 1. Comparative Example 1 and Experimental Example 1, in which the proportions of the components were confirmed, were prepared at 1 to 50% (w / w) by dissolving in ultrapure water containing no impurities.

[0100] [Table 3]

[0101] All prepared samples were placed in a high-pressure sterilizer (Jiotec, ST-105G) and heated at 121°C for 15 minutes. The heating time was measured after the target temperature was reached, regardless of the time it took for the equipment to heat up. After the heat treatment, the samples were removed and left at room temperature for 10 minutes, and then analyzed using HPLC under the conditions shown in Table 1 of Example 1.

[0102] All experiments were performed in triplicate and the results are shown in Table 4 below.

[0103] [Table 4]

[0104] As a result of the experiment, it was confirmed that in Experimental Example 1, which contained a certain amount of allulose disaccharide, the increase rate of HMF was significantly smaller. Although the heating temperature was very high and it was not possible to completely block the production of HMF, it was confirmed that stability was significantly improved compared to Comparative Example 1, in which sugar was added in place of allulose disaccharide in the same ratio as the disaccharide, where the monosaccharides directly absorbed heat damage and were rapidly decomposed. In other words, it was confirmed that allulose disaccharide has a significantly higher effect of delaying and protecting monosaccharides from deterioration due to heating compared to other disaccharides.

[0105] Furthermore, from the viewpoint of concentration, it was confirmed that the higher the absolute amount of monosaccharides, which are HMF-producing substances, the greater the HMF production under similar heating conditions. However, compared to Comparative Example 1 in which sugar was added, Experimental Example 1 in which allulose disaccharide was added produced significantly less HMF, confirming that the denaturation of monosaccharides was delayed.

[0106] This confirmed that allulose disaccharides suppress the dehydration, decomposition, and denaturation of monosaccharides even in extremely high-temperature and harsh environments, and that allulose disaccharides can be useful for suppressing the production of HMF and the dehydration of sugars.

[0107] Example 2-2: Comparison of HMF production rate depending on allulose disaccharide content The disaccharides (dimers) separated in Example 1 were mixed as shown in Table 5 below to prepare samples with different disaccharide ratios.

[0108] Specifically, crystalline allulose (CJ CheilJedang, purity 99% or higher), which has the highest composition ratio of monosaccharides, was added to ultrapure water without impurities to prepare a 10% (w / w) solution, the same as the average concentration of a typical beverage, for use as Experimental Example 2. Furthermore, allulose disaccharides with a purity of 95% or higher, isolated in Example 1, were quantitatively measured and added to the crystalline allulose, and then similarly dissolved in ultrapure water to prepare a 10% solution, which was used as Experimental Examples 3 and 4. The composition of each of the prepared samples was analyzed again using HPLC under the conditions in Table 1 of Example 1, and it was confirmed that the amount of disaccharides contained differed, as shown in Table 5 below.

[0109] [Table 5]

[0110] Each sample was heated at 95°C, the normal processing temperature for beverages, and samples were taken at 20-minute intervals to check for changes in the components and the amount of HMF produced. HMF was quantified using HPLC under the conditions shown in Table 1 of Example 1.

[0111] All experiments were performed in triplicate and the results are shown in Table 6 below.

[0112] [Table 6] * Different vertical letters a, b, c, d indicate that there is a significant difference (p<0.05) in the same sample over time.

[0113] In Experiments 2 to 4, which contained a certain amount of disaccharides, it was confirmed that a sufficient amount of disaccharides first absorbed heat damage and decomposed, resulting in an increase in monosaccharides. This confirmed that the increase in HMF generated from monosaccharides was significantly smaller. In particular, in Experiment 4, which contained the highest amount of disaccharides (2.1% (w / w) of the constituent sugars, approximately 0.21% (w / w) based on the total sample amount), it was confirmed that there was no statistically significant difference in the increase in HMF even after 60 minutes of heating, confirming the significant effect of suppressing HMF generation.

[0114] This confirmed that allulose disaccharides inhibit the dehydration, decomposition, and denaturation of monosaccharides, and inhibit the production of HMF. This indicates that allulose disaccharides significantly delay the quality deterioration phenomenon that occurs when heat causes changes in the quality of food and beverages during the normal processing and distribution process, resulting in the production of HMF.

[0115] Through this experimental process, it was confirmed that when a certain amount of disaccharides containing allulose is contained, the decomposition and denaturation of sugars (allulose) by heat, resulting in the production of HMF, can be significantly delayed.

[0116] Compared to conventional methods that use additives consisting of completely different components (e.g., additives such as antioxidants) to inhibit HMF production, this method uses disaccharides based on sugars, which has the advantage of having very little impact on the taste and characteristics of the product.

[0117] Example 3: Identification of the structure of allulose disaccharides To confirm the structure of allulose disaccharides having the function of inhibiting HMF production, the structures of D1 and D2 separated in Example 1 were identified through ESI-MS, 1 H NMR, and 13 C NMR.

[0118] Specifically, the structure was identified by the following method.

[0119] Major 6-O-β-D-Psicopyranosyl-α-D-psicofuranose is a white amorphous powder, ESI-MS m / z 365 [M+Na]+; 1H NMR (850 MHz, D2O) δH 3.44 (1H, d, J = 12.0 Hz), 3.47 (1H, d, J = 12.0 Hz), 3.56 (1H, dd, J = 11.0, 5.0 Hz), 3.60 (1H, d, J = 12.0 Hz), 3.62 (1H, dd, J = 11.0, 2.5 Hz), 3.70 (1H, br d, J = 12.5 Hz), 3.75 (1H, d, J = 12.0 Hz), 3.75 (1H, br ma), 3.82 (1H, br d, J = 12.5 Hz), 3.84 (1H, br s), 3.92 (1H, t, J = 3.0 Hz), 3.97 (1H, d, J = 5.5 Hz), 4.09 (1H, t, J = 5.5 Hz), 4.13 (1H, br m) [D2O signal δH 4.70]; 13C NMR signalsb δC 57.6, 60.4, 62.9, 64.7, 64.9, 69.1, 68.9, 70.2, 70.3, 81.2, 101.8, 103.4.

[0120] Minor 6-O-β-D-Psicopyranosyl-β-D-psicofuranose is a white amorphous powder, ESI-MS m / z 365 [M+Na]+; 1H NMR (850 MHz, D2O) δH 3.49 (1H, d, J = 13.0 Hz), 3.73 (1H, d, J = 13.0 Hz), 3.58 (1H, ma), 3.68 (1H, dd, J = 11.0, 2.5 Hz), 3.62 (1H, ma), 3.71 (1H, br d, J = 12.0 Hz), 3.82 (1H, br d, J = 12.0 Hz), 3.76 (1H, br ma), 3.78 (1H, ma), 3.87 (1H, br s), 3.98 (1H, t, J = 3.0 Hz), 3.95 (1H, d, J = 4.5 Hz), 4.00 (1H, br m), 4.34 (1H, dd, J = 8.0, 4.5 Hz) [D2O signal δH 4.70]; 13C NMR signalsb δC 57.7, 61.4, 62.2, 64.7, 64.8, 69.0, 69.2, 70.8, 74.4, 80.8, 101.8, 105.9.

[0121] As a result, it was confirmed that D1 is a novel allulose disaccharide and has the structure of the following chemical formula (1).

[0122] [ka] ···(1)

[0123] Furthermore, D1 has two forms, major and minor (Figure 3). The major form, 6-O-β-D-Psicopyranosyl-α-D-psicofuranose, has the structure shown in chemical formula (2) below, and the minor form, 6-O-β-D-Psicopyranosyl-β-D-psicofuranose, has the structure shown in chemical formula (3) below.

[0124] [ka] ···(2)

[0125] [ka] ···(3)

[0126] The compound of chemical formula (2) (6-O-β-D-Psicopyranosyl-α-D-psicofuranose) was named Compound A, and the compound of chemical formula (3) (6-O-β-D-Psicopyranosyl-β-D-psicofuranose) was named Compound B.

[0127] Meanwhile, D2 was confirmed to be a novel allulose disaccharide in which, in relation to the chemical formula (1) and a structural isomer, the hydroxyl group at the 2nd carbon (C2; according to the carbon numbering in Figure 4) of allulose is glycosidic bonded to the hydroxyl group at any one of the 1st to 6th carbons (C1 to C6) of another allulose molecule.

[0128] From the above description, those skilled in the art to which the present application pertains will understand that the present application may be implemented in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the above-described embodiments are merely illustrative and not limiting. The scope of the present application should be interpreted as including within the meaning and scope of the claims below, and any modifications or variations derived from the equivalent concepts thereof, rather than the above detailed description.

Claims

1. Use of allulose disaccharide for inhibiting the production of hydroxymethylfurfural (HMF), The allulose disaccharide is a compound in which two allulose molecules are linked by a glycosidic bond, and the glycosidic bond is formed by a glycosidic bond between the hydroxyl group at the 2nd carbon (C2) of one allulose molecule and the hydroxyl group at any one of the 1st to 6th carbons (C1 to C6) of the other allulose molecule.

2. The use according to claim 1, wherein the HMF production is due to sugars.

3. The use according to claim 2 , wherein the sugar is a monosaccharide.

4. The use according to claim 3, wherein the monosaccharide is allulose.

5. The allulose disaccharide has the following chemical formula (1): 【Chemical 1】 （１） The use according to claim 1, wherein

6. Use of allulose disaccharides for preventing non-enzymatic browning, The allulose disaccharide is a compound in which two allulose molecules are linked by a glycosidic bond, and the glycosidic bond is formed by a glycosidic bond between the hydroxyl group at the 2nd carbon (C2) of one allulose molecule and the hydroxyl group at any one of the 1st to 6th carbons (C1 to C6) of the other allulose molecule.

7. The use according to claim 6, wherein the non-enzymatic browning is due to sugars.

8. The use according to claim 7 , wherein the sugar is a monosaccharide.

9. The use according to claim 8, wherein the monosaccharide is allulose.

10. The allulose disaccharide has the following chemical formula (1): 【Chemistry 2】 （１） The use according to claim 6, wherein

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