Method for producing steviol glycoside composition

Abstract

Provided are a novel method for producing a steviol glycoside composition and a method for increasing the concentration of a steviol glycoside in a steviol glycoside-containing solution. In addition, the present invention provides a food or drink containing the steviol glycoside composition produced by the production method. The present invention is a method for producing a steviol glycoside composition, characterized by comprising preparing a steviol glycoside-containing solution containing a stevia extract, preparing a treatment liquid by adding an additive to the steviol glycoside-containing solution, and adding the additive such that the pH of the treatment liquid reaches 10-13.

Description

[0001] This application is a divisional application of patent application No. 202180044864.1, filed on April 23, 2021, entitled "Method for manufacturing steviol glycoside composition". Technical Field

[0002] This invention relates to a method for manufacturing a steviol glycoside composition and a method for increasing the steviol glycoside concentration in a solution containing steviol glycosides. Furthermore, this invention also relates to a food or beverage containing the steviol glycoside composition manufactured by the aforementioned method. Background Technology

[0003] The leaves of stevia rebaudiana, a member of the Asteraceae family, contain a secondary metabolite called steviol, a diterpenoid. Steviol glycosides are used in the food industry as a low-calorie sweetener due to their approximately 300 times sweetness of sugar. With obesity becoming a serious social problem internationally, the demand for low-calorie sweeteners is increasing from the perspective of improving health and reducing medical costs. Currently, synthetic amino acid derivatives such as aspartame and acesulfame potassium are used as artificial sweeteners; however, naturally occurring low-calorie sweeteners like steviol glycosides are considered safer and are expected to gain public acceptance.

[0004] Various compositions containing steviol glycosides have been reported to date. For example, Patent Document 1 describes a method for extracting steviol glycoside compositions from the leaves of Stevia rebaudiana plants containing rebaudioside M at a higher concentration than the common relative concentration.

[0005] Patent documents

[0006] Patent Document 1: International Publication No. 2017 / 035527 Summary of the Invention

[0007] This invention provides a novel method for manufacturing a steviol glycoside composition and a method for increasing the concentration of steviol glycosides in a solution containing steviol glycosides. Furthermore, this invention provides a food or beverage containing the steviol glycoside composition manufactured by the aforementioned method.

[0008] The inventors conducted in-depth research and discovered that, in the process of producing steviol glycosides from dried leaves of the stevia plant, by adjusting the pH to a specified range, the concentration of the steviol glycoside composition in the solution containing steviol glycosides can be increased, thereby completing the present invention. The present invention is as follows. [1]

[0010] A method for manufacturing a steviol glycoside composition, characterized in that,

[0011] This includes preparing a solution containing steviol glycosides derived from stevia extract.

[0012] And to prepare the first treatment solution (or simply the "treatment solution" if no second treatment solution is added) by adding a first additive (when no further additives are added) to the solution containing steviol glycosides.

[0013] The first additive is added in such a way that the pH of the first treatment solution reaches 10-13.

[0014] [1A]

[0015] A method for manufacturing a steviol glycoside composition, characterized in that,

[0016] This includes preparing a solution containing steviol glycosides derived from stevia extract.

[0017] Additives are added to the solution containing steviol glycosides to prepare the treatment solution.

[0018] The additive is added to achieve a pH of 10-13 in the treated solution. [2]

[0020] According to the manufacturing method described in [1], the first additive is characterized in that it is an alkali agent.

[0021] [2A]

[0022] The manufacturing method according to [1A] is characterized in that the additive is an alkali agent. [3]

[0024] The manufacturing method according to [2] or [2A] is characterized in that the alkali contains one or more compounds selected from alkali metal hydroxides, alkaline earth metal hydroxides, silicates, carbonates and bicarbonates. [4]

[0026] The manufacturing method according to any one of [1] to [3] is characterized in that the steviol glycoside composition contains one or more steviol glycosides selected from ribobadiin A, ribobadiin B, ribobadiin M, ribobadiin C, ribobadiin D, steviol glycoside, ribobadiin F, dukorbitol A, ribobadiin G, ribobadiin N, raspberry glycoside and steviol disaccharide glycoside. [5]

[0028] The manufacturing method according to any one of [1] to [4] is characterized in that, after preparing the first treatment liquid, the first treatment liquid is kept for 1 minute to 3 days.

[0029] [5A]

[0030] The manufacturing method according to any one of [1] to [4] is characterized in that, after preparing the treatment liquid, the treatment liquid is kept for 1 minute to 3 days. [6]

[0032] The manufacturing method according to any one of [1] to [5] is characterized in that,

[0033] The process further includes adding a second additive to the first treatment solution to prepare a second treatment solution.

[0034] The second additive is added in such a way that the pH of the second treatment solution reaches 2 to 10.

[0035] [6A]

[0036] The manufacturing method according to any one of [1] to [5A] is characterized in that,

[0037] The process further includes adding a second additive to the treatment solution to prepare a second treatment solution.

[0038] The second additive is added in such a way that the pH of the second treatment solution reaches 2 to 10. [7]

[0040] According to the manufacturing method described in [6], the characteristic is that one or more compounds selected from aluminum sulfate, aluminum polychloride, ferric chloride (III) or its hydrates, polyacrylamide hydrolysate, alginic acid, chitosan and chitosan are further added to the first treatment liquid and / or the second treatment liquid.

[0041] [7A]

[0042] According to the manufacturing method described in [6A], the method is characterized in that one or more compounds selected from aluminum sulfate, aluminum polychloride, ferric chloride (III) or its hydrates, polyacrylamide hydrolysate, alginic acid, chitosan and chitosan are further added to the treatment liquid and / or the second treatment liquid. [8]

[0044] A method for increasing the concentration of steviol glycosides in a solution containing steviol glycosides, characterized in that,

[0045] This includes preparing a solution containing steviol glycosides derived from stevia extract.

[0046] The first treatment solution is prepared by adding the first additive to the solution containing steviol glycosides.

[0047] The first additive is added in such a way that the pH of the first treatment solution reaches 10-13.

[0048] [8A]

[0049] A method for increasing the concentration of steviol glycosides in a solution containing steviol glycosides, characterized in that,

[0050] This includes preparing a solution containing steviol glycosides derived from stevia extract.

[0051] Additives are added to the solution containing steviol glycosides to prepare the treatment solution.

[0052] The additive is added to achieve a pH of 10-13 in the treated solution. [9]

[0054] The method according to [8] or [8A] is characterized by increasing the concentration of one or more steviol glycosides selected from ribobadiin A, ribobadiin C, steviol glycoside, ribobadiin F, ribobadiin G and raspberry glycoside.

[10]

[0056] A steviol glycoside composition, characterized in that it is manufactured by any one of the methods described in [1] to [7A].

[11]

[0058] A food product characterized by containing the steviol glycoside composition described in

[10] .

[0059] According to the present invention, a novel method for manufacturing a steviol glycoside composition is provided. Furthermore, according to the present invention, a method for increasing the concentration of the steviol glycoside composition in a solution containing steviol glycosides is provided. Attached Figure Description

[0060] Figure 1 A graph showing the changes in the concentration of steviol glycosides in a solution containing steviol glycosides.

[0061] Figure 2 This is a graph showing the change in RebA concentration as the environment changes from alkaline to neutral.

[0062] Figure 3 A graph showing the concentration variation of RebA based on the type of alkali agent. Detailed Implementation

[0063] The present invention will now be described in detail. The following embodiments are illustrative of the invention and are not intended to limit the invention to these embodiments only. The present invention can be implemented in various ways without departing from its spirit. Furthermore, all documents, publications, patent gazettes, and other patent documents cited in this specification are incorporated herein by reference.

[0064] In this manual, “rebaudioside”, “Reb”, and “Reb.” have the same meaning and all refer to “rebaudioside”.

[0065] 1. Method for manufacturing steviol glycoside composition

[0066] The method for manufacturing the steviol glycoside composition of the present invention (hereinafter also referred to as "the manufacturing method of the present invention") comprises preparing a solution containing steviol glycosides of stevia extract (hereinafter also referred to as "the solution containing steviol glycosides"), and adding a first additive to the solution containing steviol glycosides to prepare a first treatment solution, and adding the first additive in such a way that the first treatment solution reaches a pH of 10 to 13.

[0067] Furthermore, the manufacturing method of the present invention includes preparing a solution containing steviol glycosides of stevia extract, and adding an additive to the solution containing steviol glycosides to prepare a treatment solution, and adding the additive in such a way that the treatment solution reaches a pH of 10 to 13.

[0068] According to the manufacturing method of the present invention, by adjusting the pH of the solution containing steviol glycosides to 10-13, the concentration of specific steviol glycosides in the steviol glycoside composition obtained as a result can be increased.

[0069] Stevia leaves contain a variety of components besides steviol glycosides. When preparing a steviol glycoside composition with steviol extract as the main component, the components other than steviol glycosides in the composition are considered to be substantially derived from stevia, provided no other components are arbitrarily added. These components derived from stevia, besides steviol glycosides, include water-soluble and insoluble components. Examples of water-soluble components include water-soluble dietary fiber and other polysaccharides, alkaloids and secondary metabolites such as flavonoids and terpenoids, methanol, polyphenols, minerals, vitamins, amino acids, organic acids, water-soluble proteins, and various other glycosides. Examples of insoluble components include insoluble polysaccharides including insoluble dietary fiber, insoluble proteins, and lipids.

[0070] Although not bound by theory, it is generally believed that in a manufacturing method according to one aspect of the present invention, steviol glycosides bound to certain components from stevia dissociate from the binding due to an increase in pH, thereby increasing the amount of purified glycosides and ultimately increasing the concentration in the resulting steviol glycoside composition.

[0071] In this specification, the term "increased concentration of steviol glycosides" refers to an increase in the concentration of steviol glycosides compared to the concentration in a steviol glycoside composition obtained without adjusting the steviol glycoside-containing solution to the specified pH. That is, the increase in concentration only needs to be relative and does not require the steviol glycoside concentration to reach a specific value. Furthermore, the increase in steviol glycoside concentration can be expressed as an increase rate (%) based on the aforementioned comparative control.

[0072] The concentration of steviol glycosides can be determined by LC / MS or HPLC.

[0073] The steviol glycoside composition obtained by the manufacturing method of the present invention comprises one or more steviol glycosides selected from ribobadiin A, ribobadiin B, ribobadiin M, ribobadiin C, ribobadiin D, steviol glycoside, ribobadiin F, dukorbitol A, ribobadiin G, ribobadiin N, raspberry glycoside, and steviol disaccharide glycoside.

[0074] Furthermore, the steviol glycoside composition obtained by the manufacturing method of the present invention may, in addition to the steviol glycosides listed above, further contain, for example, one or more steviol glycosides selected from ribobadiin E, ribobadiin I, ribobadiin J, ribobadiin K, ribobadiin M, ribobadiin O, ribobadiin Q, ribobadiin R, durqueside C, steviol, and steviol monosaccharides.

[0075] The steviol glycoside composition obtained by the manufacturing method of the present invention is a substance selected from the above-listed steviol glycosides, namely, ribobadiin A, ribobadiin C, steviol glycoside, ribobadiin F, ribobadiin G and raspberry glycoside, with an increased concentration of one or more of them.

[0076] In one embodiment of the present invention, the increase rate of the concentration of ribobandi glycoside A when the first treatment solution or the treatment solution is adjusted to a specified pH may also be 101-150%, 101-140%, 103-140%, 105-140%, 110-140%, 115-140%, 120-140%, 121-140%, 122-140%, 123-140%, 124-140%, 125-140%, 126-140%, 127-140%, 128-140%, 129-140%, or 130-140%, etc.

[0077] In one embodiment of the present invention, the increase rate of the concentration of ribobandi glycoside C when the first treatment solution or the treatment solution is adjusted to a specified pH may also be 101-170%, 101-165%, 101-160%, 105-160%, 110-160%, 115-160%, 120-160%, 125-160%, 130-160%, 131-160%, 132-160%, 133-160%, 134-160%, 135-160%, 136-160%, 137-160%, 138-160%, 139-160%, or 140-160%, etc.

[0078] In one embodiment of the present invention, the increase rate of steviol glycoside concentration when the first treatment solution or the treatment solution is adjusted to a specified pH may also be 101-200%, 101-190%, 101-185%, 101-180%, 110-180%, 120-180%, 125-180%, 130-180%, 135-180%, 140-180%, 145-180%, 150-180%, 151-180%, 152-180%, 153-180%, 154-180%, 155-180%, 156-180%, 157-180%, 158-180%, 159-180%, or 160%-180%, etc.

[0079] In one embodiment of the present invention, the increase rate of the concentration of riboside F when the first treatment solution or the treatment solution is adjusted to a specified pH may also be 101-150%, 101-140%, 103-140%, 105-140%, 110-140%, 115-140%, 120-140%, 121-140%, 122-140%, 123-140%, 124-140%, 125-140%, 126-140%, 127-140%, 128-140%, 129-140%, or 130-140%, etc.

[0080] In one embodiment of the present invention, the increase rate of the concentration of riboside G when the first treatment solution or the treatment solution is adjusted to a specified pH may be 101-350%, 101-340%, 101-330%, 110-330%, 120-330%, 130-330%, 140-330%, 150-330%, 160-330%, 170-330%, 180-330%, 190-330%, 200-330%, 210-330%, 220-330%, 230-330%, 240-330%, or 250-330%, etc.

[0081] In one embodiment of the present invention, the increase rate of the concentration of raspberry glycoside in the first treatment solution or the treatment solution adjusted to a specified pH may be 101-700%, 101-690%, 101-680%, 101-670%, 110-670%, 120-670%, 130-670%, 140-670%, 150-670%, 160-670%, 170-670%, 180-670%, 190-670%, or 200-670%, etc.

[0082] In one embodiment of the present invention, the increase rate of the concentration of each of the above-mentioned steviol glycosides refers to the increase rate when a solution containing steviol glycosides is obtained by extracting stevia extract once with 30 times the amount of ion-exchanged water (60℃±5℃) containing dried stevia leaves (moisture content: 3~4% by weight) used as raw material.

[0083] (A) Prepare a solution containing steviol glycosides from stevia extract.

[0084] The manufacturing method of the present invention includes preparing a solution containing steviol glycosides of stevia extract. As a method for obtaining stevia extract, for example, extracting dried leaves of the stevia plant using an aqueous solvent to obtain an extract (extract).

[0085] In this specification, the term "dried leaves of stevia" refers to leaves whose moisture content has been reduced by drying fresh leaves of stevia. The moisture content of the dried leaves of stevia is preferably 1 to 10% by weight, more preferably 2 to 8% by weight, and particularly preferably 3 to 4% by weight.

[0086] Steviosides can be extracted from dried leaves using solvents such as water, alcohol, or a mixture thereof. Preferred extraction solvents include ion-exchanged water, pure water (e.g., MilliQ water), and aqueous ethanol solutions. The dried leaves may or may not be pulverized during extraction. Pulverization can be performed using a ball mill or similar device. Alternatively, a kneading extractor (SKN-R100, manufactured by Sanyu Machinery Co., Ltd.) can be used for extraction.

[0087] Heating the aqueous solvent during extraction allows for more efficient extraction of steviol glycosides. Extraction temperatures can be, for example, 25–80°C, 30–75°C, 35–70°C, 40–65°C, or 45–70°C, with 45–70°C being preferred. Furthermore, the lower and upper limits of the temperature specified in this specification can be, for example, ±1°C, ±2°C, ±3°C, ±4°C, or ±5°C.

[0088] Extraction can be performed multiple times, not just once. Multiple extractions can extract more steviol glycosides from the leaves. From an efficiency standpoint, approximately two extractions are preferable.

[0089] In one embodiment of the present invention, the solution containing steviol glycosides may be the steviol extract itself obtained by the above method. Furthermore, the solution containing steviol glycosides need only contain steviol extract, and a steviol glycoside composition produced by any other method (e.g., using steviol glycoside hydrolase, etc.) may also be added.

[0090] Furthermore, in one embodiment of the present invention, the obtained stevia extract may also be subjected to solid-liquid separation treatment. As for solid-liquid separation treatment, there are no particular limitations as long as the solid and liquid can be sufficiently separated; examples include treatment using a centrifuge or filter press, or gravity filtration using a filter or sieve.

[0091] Solid-liquid separation can also be performed using various methods; for example, a second solid-liquid separation process can be performed after the first solid-liquid separation process.

[0092] (B) Adding the first additive or additive to a solution containing steviol glycosides to prepare the first treatment solution or treatment solution.

[0093] The manufacturing method of the present invention includes adding a first additive to a solution containing steviol glycosides prepared according to method (A) above to prepare a first treatment solution. Furthermore, the manufacturing method of the present invention includes adding an additive to a solution containing steviol glycosides prepared according to method (A) above to prepare a treatment solution. In one embodiment of the present invention, the first additive or additive is an alkaline agent. Examples of alkaline agents include alkali metal hydroxides, alkaline earth metal hydroxides, silicates, carbonates, bicarbonates, etc.

[0094] Examples of alkali metal hydroxides include sodium hydroxide and potassium hydroxide. Examples of alkaline earth metal hydroxides include calcium hydroxide and magnesium hydroxide. Examples of silicates include sodium metasilicate, sodium orthosilicate, potassium metasilicate, and potassium orthosilicate. Examples of carbonates include sodium carbonate and potassium carbonate. Examples of bicarbonates include sodium bicarbonate and potassium bicarbonate. Among these, sodium hydroxide, potassium hydroxide, and calcium hydroxide are particularly preferred. Furthermore, these alkaline agents can be used alone or in combination of two or more.

[0095] In a preferred embodiment of the present invention, the first additive is added to the solution containing steviol glycosides such that the pH of the first treatment solution reaches 10-13. That is, the amount of the first additive is not particularly limited, as long as the pH of the first treatment solution reaches the aforementioned range; it can be added while appropriately adjusting the pH of the first treatment solution to reach 10-13, depending on the amount of the steviol glycoside-containing solution.

[0096] Furthermore, in one embodiment of the present invention, the pH of the first treatment solution or the treatment solution may also be pH 10-13, pH 10-12.5, pH 10.1-12.5, pH 10.2-12.5, pH 10.3-12.5, pH 10.4-12.5, pH 10.5-12.5, pH 10.6-12.5, pH 10.7-12.5, pH 10.8-12.5, pH 10.9-12.5, pH 11-13, pH 11-12.5, pH 11.1-12.5, pH 11.2-12.5, pH pH 11.3~12.5, pH 11.4~12.5, pH 11.5~12.5, pH 11.6~12.5, pH 11.7~12.5, pH 11.8~12.5, pH 11.9~12.5, pH 11~12.2, pH 11.1~12.2, pH 11.2~12.2, pH 11.3~12.2, pH 11.4~12.2, pH 11.5~12.2, pH 11.6~12.2, pH 11.7~12.2, pH 11.8~12.2, or pH 11.9~12.2, etc.

[0097] The temperature of the solution containing steviol glycosides when the first additive or additive is added can also be 1~60℃, 5~60℃, 10~60℃, 15~60℃, 20~60℃, 25~60℃, 30~60℃, 40~60℃, 50~60℃, 1~50℃, 5~50℃, 10~50℃, 15~50℃, 20~50℃, 25~50℃, 30~50℃, 1~40℃, 5~40℃, 10~40℃, 15~40℃, 20~40℃, 25~40℃, 30~40℃, 1~30℃, 5~30℃, 10~30℃, 15~30℃, or 20~30℃, etc.

[0098] Furthermore, in one embodiment of the present invention, the first treatment solution or treatment liquid may also be maintained for a predetermined time before the preparation of the second treatment solution described later. In this specification, "maintaining the first treatment solution or treatment liquid" means maintaining a predetermined pH value or range after the preparation of the first treatment solution or treatment liquid and between the addition of the second additive for the preparation of the second treatment solution described later. The first treatment solution or treatment liquid may be maintained without any treatment, or it may be maintained based on the implementation of some treatments.

[0099] Examples of the time for maintaining the first treatment solution or the treatment solution include 1 minute to 3 days, 1 minute to 2 days, 1 minute to 24 hours, 1 minute to 12 hours, 1 minute to 10 hours, 1 minute to 8 hours, 1 minute to 6 hours, 1 minute to 4 hours, 1 minute to 3 hours, 1 minute to 2 hours, 2 minutes to 2 hours, 3 minutes to 2 hours, 4 minutes to 2 hours, 5 minutes to 2 hours, 10 minutes to 2 hours, 15 minutes to 2 hours, 20 minutes to 2 hours, 25 minutes to 2 hours, 30 minutes to 2 hours, 35 minutes to 2 hours, 40 minutes to 2 hours, 45 minutes to 2 hours, 50 minutes to 2 hours, 55 minutes to 2 hours, or 1 to 2 hours. According to the manufacturing method of the present invention, by adding the first additive or adjusting the pH to a range of 10 to 13 to a solution containing steviol glycosides, the concentration of specific steviol glycosides in the steviol glycoside composition is increased. The first treatment solution or solution, which was once adjusted to a pH range of 10-13, remained stable even after the addition of the second additive, without any decrease in the concentration of steviol glycosides.

[0100] Maintain the temperature of the first treatment solution or the treatment solution, for example, room temperature (approximately 25°C).

[0101] (C) Prepare a second treatment solution by adding the second additive to the first treatment solution or the treatment solution.

[0102] The manufacturing method of the present invention further includes adding a second additive to a first treatment solution to prepare a second treatment solution. Furthermore, the manufacturing method of the present invention further includes adding a second additive to the treatment solution to prepare a second treatment solution. In one embodiment of the present invention, the second additive is an additive that has the function of lowering pH. Such an additive only needs to have the function of lowering pH, and any substance can be used, such as organic pH adjusters like citric acid, lactic acid, and acetic acid, and inorganic pH adjusters like phosphoric acid, hydrochloric acid, sulfuric acid, and carbon dioxide. In addition, compounds such as aluminum sulfate, aluminum polychloride, ferric chloride (III), or their hydrates, described later, can also be used. These second additives can be used alone or in combination of two or more.

[0103] In a preferred embodiment of the present invention, the second additive is added to the first treatment solution or treatment solution such that the pH of the second treatment solution reaches 2 to 10. That is, the amount of the second additive added is not particularly limited, as long as the pH of the second treatment solution reaches the aforementioned range. It can be added while appropriately adjusting the amount of the first treatment solution or treatment solution to achieve a pH of 2 to 10 in the second treatment solution. Furthermore, when using compounds such as aluminum sulfate, aluminum polychloride, ferric chloride (III), or their hydrates, the coagulation reaction described later should also be considered, and the amount added should be adjusted accordingly.

[0104] According to the manufacturing method of the present invention, by adding a first additive or adjusting the pH to a range of 10-13 to a solution containing steviol glycosides, the concentration of a specific steviol glycoside in the steviol glycoside composition increases. Even after adjusting the pH to a range of 2-10 with a second additive, the increased concentration of steviol glycosides remains stable and does not decrease after the first treatment solution or treatment solution has been adjusted to a pH range of 10-13.

[0105] In addition, in one embodiment of the present invention, the second treatment solution may also be pH 2~10, pH 2.5~9.5, pH 3~9, pH 3.5~8.5, pH 4~8, pH 4.5~8 or pH 5~8, etc.

[0106] The temperature at which the second additive is added to the first treatment solution or the treatment solution may be, for example, room temperature (approximately 25°C).

[0107] In a preferred embodiment of the present invention, one or more compounds selected from aluminum sulfate, aluminum polychloride, ferric chloride (III) or its hydrates, polyacrylamide hydrolysate, alginate, chitosan, and chitin may be further added to the first treatment solution or the treatment solution and / or the second treatment solution. It is sufficient to contain one or more of these compounds, or a combination of two or more. Furthermore, these compounds may be added during step "B" as described above, or during step "C" as described above, or in both steps "B" and step "C".

[0108] The amount of the above-mentioned compound added is not particularly limited, as long as it causes a coagulation reaction in the first treatment solution or the second treatment solution. For example, it can be added in an amount of 3.0 to 50% by weight relative to the soluble solids content in the treatment solution. For example, when it is ferric chloride (III) hexahydrate, it can be added in an amount equivalent to 15 to 40% by weight of the solids content in the treatment solution, preferably 18 to 38% by weight, and more preferably 20 to 35% by weight. When it is a 0.5% (w / v) chitosan solution, it can be added in an amount equivalent to 3.0 to 10% by weight of the soluble solids content in the treatment solution, preferably 4.0 to 8.0% by weight, and more preferably 4.5 to 7.0% by weight.

[0109] Furthermore, the pH during coagulation treatment can be appropriately selected based on the types of the aforementioned compounds to optimize coagulation. In one embodiment of the present invention, the pH of the first treatment solution or the treatment solution and / or the second treatment solution during coagulation treatment may also be pH 2-13, pH 3-13, pH 4-13, pH 5-13, or pH 6-13, etc.

[0110] Furthermore, the coagulation process can be carried out at room temperature (approximately 25°C) without heating or cooling.

[0111] In one embodiment of the present invention, the coagulants contained in the first treatment liquid or the treatment liquid and / or the second treatment liquid may also be removed after the above-described coagulation treatment and before any of the resin purification treatments described later. The removal of the coagulants can be carried out by known methods such as filtration.

[0112] The manufacturing method of the present invention may, as desired, further subject the second processing liquid to the following treatments (D) to (F).

[0113] (D) Resin purification treatment

[0114] In one embodiment of the present invention, the second treatment solution prepared after the above treatments can also be treated with a hydrophobic porous resin. Stevioside is amphiphilic, having both hydrophilic and hydrophobic groups in its molecular structure, and has a molecular weight of approximately 1,000. Furthermore, it is known to be stable at pH 2.5–9.0 and does not ionize under acidic or alkaline conditions. On the other hand, the first treatment solution or treatment solution after coagulation treatment also contains a large amount of components other than steviol glycosides. Although not theoretically constrained, it is generally believed that these components include those with molecular weights different from steviol glycosides, such as iron ions, or those that are generally ionized like amino acids; these components can be removed by treatment with a hydrophobic porous resin.

[0115] Steviosides, possessing a hydrophobic steviol backbone, are captured by hydrophobic bonding with synthetic resin. Conversely, highly hydrophilic impurities do not bond with the resin but migrate to the permeate and are thus removed. Therefore, it is believed that introducing a coagulated treatment solution into a column filled with the aforementioned resin, followed by washing with water, can improve the purity of steviol glycosides. Furthermore, the bonding between steviol glycosides and the functional groups of the synthetic resin is dissociated by a low-polarity solvent, thus offering the advantage of ultimately achieving high-yield recovery of steviol glycosides.

[0116] The hydrophobic porous resin used in the manufacturing method of one embodiment of the present invention is not particularly limited, as long as it is a porous resin with low affinity for water. For example, a porous resin selected from one or more hydrophobic resins chosen from copolymers of styrene and divinylbenzene, polyethylene, polypropylene, polystyrene, poly(meth)acrylonitrile, polyamide, and polycarbonate is preferred. In a preferred embodiment of the present invention, it is preferable that the copolymer of styrene and divinylbenzene has not undergone ion exchange group introduction treatment (i.e., it does not have ion exchange groups). Generally, when manufacturing ion exchange resin, styrene and divinylbenzene are copolymerized to form a three-dimensional network structure, and then ion exchange groups are introduced into the resin. However, "without ion exchange group introduction treatment" means that such treatment has not been performed.

[0117] In one embodiment of the present invention, the hydrophobic porous resin has a hydrophobic group, which comprises one or more selected from aryl, alkyl, alkylsilyl, ester, and epoxy groups. In one embodiment of the present invention, it is sufficient to contain one or more hydrophobic groups selected from these groups, and other hydrophobic groups may also be included. Examples of aryl groups include phenyl, benzyl, tolyl, and xylyl; examples of alkyl groups include C1-20 alkyl groups, such as methyl, ethyl, propyl, butyl, pentyl, and octadecyl.

[0118] In a preferred embodiment of the invention, the modal pore radius of the hydrophobic porous resin is 10-200 Å. In another preferred embodiment, the modal pore radius is 10-150 Å, 15-100 Å, or 20-80 Å. It is generally believed that by possessing such microporous characteristics, steviol glycosides can be effectively adsorbed into the micropores and effectively separated from other components.

[0119] Alternatively, the treatment solution can be further treated with anion exchange resin before treatment with a hydrophobic porous resin. Pretreatment with anion exchange resin effectively removes components such as pigments or catechins that are bonded to the hydrophobic resin. There are no particular limitations on the type of anion exchange resin used; for example, basic anion exchange resins can be used. Such basic anion exchange resins can be weakly basic anion exchange resins with 1-2 amino groups as functional groups, or strongly basic anion exchange resins with quaternary ammonium groups (e.g., trimethylammonium or dimethylethanolammonium).

[0120] (E) Concentration Process

[0121] The solution purified by resin can be further concentrated to remove the aqueous solvent. Such treatment is not particularly limited; examples include evaporating the aqueous solvent by heating or removing it by vacuum drying.

[0122] (F) Any additional process

[0123] Following the manufacturing method of the present invention, a crystallization process can be added to produce a high-purity (purity of 95% or more) steviol glycoside composition.

[0124] 2. Methods to increase the concentration of steviol glycosides in solutions containing steviol glycosides

[0125] The present invention also relates to a method for increasing the concentration of steviol glycosides in a solution containing steviol glycosides (hereinafter also referred to as "the method of the present invention"). The method of the present invention includes preparing a solution containing steviol glycosides and stevia extract, and adding a first additive to the solution containing steviol glycosides to prepare a first treatment solution, wherein the first additive is added in such a way that the first treatment solution reaches a pH of 10 to 13.

[0126] Furthermore, the method of the present invention includes preparing a solution containing steviol glycosides of stevia extract, and adding an additive to the solution containing steviol glycosides to prepare a treatment solution, wherein the additive is added in such a way that the treatment solution reaches a pH of 10 to 13.

[0127] Furthermore, the method of the present invention increases the concentration of one or more steviol glycosides selected from ribobadiin A, ribobadiin C, steviol glycoside, ribobadiin F, ribobadiin G and raspberry glycoside.

[0128] Furthermore, the method of the present invention has essentially the same structure as the method for manufacturing the steviol glycoside composition of the present invention. Therefore, the above description of "1. Method for manufacturing the steviol glycoside composition" also applies to the method of the present invention.

[0129] 3. Stevioside composition

[0130] The present invention also relates to a steviol glycoside composition (hereinafter also referred to as "the steviol glycoside composition of the present invention") manufactured by the manufacturing method of the present invention. In one embodiment of the present invention, the steviol glycoside composition can also be used as a sweetener composition.

[0131] When used as a sweetener composition, in addition to the steviol glycoside composition of the present invention, sweeteners other than steviol glycosides may also be included. Examples of such sweeteners include natural sweeteners such as fructose, sucrose, glucose-fructosesyrup, glucose, maltose, high fructose syrup, sugar alcohols, oligosaccharides, honey, sugarcane juice (molasses), maltose, monk fruit powder, monk fruit extract, licorice powder, licorice extract, arrowroot seed powder, arrowroot seed extract, etc., or artificial sweeteners such as acesulfame potassium, sucralose, neotame, aspartame, saccharin, etc. From the viewpoints of refreshingness, ease of drinking, natural taste, and imparting a moderate alcoholic aroma, natural sweeteners are preferred, and fructose, glucose, maltose, sucrose, and saccharin are particularly suitable. Only one of these sweeteners may be used, or multiple sweeteners may be used.

[0132] 4. Foods and beverages containing steviol glycosides

[0133] This invention also relates to a food and beverage containing a steviol glycoside composition obtained by the manufacturing method of this invention (hereinafter also referred to as "the food and beverage of this invention"). The food and beverage of this invention is not particularly limited as long as it contains the steviol glycoside composition of this invention. Here, "food and beverage" refers to beverages and food products. In a preferred embodiment, the food and beverage is a beverage.

[0134] The amount of steviol glycosides of the present invention contained in the food and beverage products of the present invention varies depending on the specific food and beverage product. In the case of a beverage, it is preferably about 1 ppm to 800 ppm by mass, for example, it may also be 20 ppm to 750 ppm by mass, 20 ppm to 700 ppm by mass, 20 ppm to 650 ppm by mass, 20 ppm to 600 ppm by mass, 20 ppm to 550 ppm by mass, 25 ppm to 550 ppm by mass, 30 ppm to 550 ppm by mass, 35 ppm to 550 ppm by mass, 40 ppm to 550 ppm by mass, 45 ppm to 550 ppm by mass, or 50 ppm by mass. 550 ppm, 55 ppm to 550 ppm, 20 ppm to 540 ppm, 25 ppm to 540 ppm, 30 ppm to 540 ppm, 35 ppm to 540 ppm, 40 ppm to 540 ppm, 45 ppm to 540 ppm, 50 ppm to 540 ppm, 55 ppm to 540 ppm, 20 ppm to 530 ppm, 25 ppm to 530 ppm, 30 ppm to 530 ppm, 35 ppm to 530 ppm, 40 ppm to 530 ppm 45 ppm to 530 ppm, 50 ppm to 530 ppm, 55 ppm to 530 ppm, 20 ppm to 520 ppm, 25 ppm to 520 ppm, 30 ppm to 520 ppm, 35 ppm to 520 ppm, 40 ppm to 520 ppm, 45 ppm to 520 ppm, 50 ppm to 520 ppm, 55 ppm to 520 ppm, 20 ppm to 510 ppm, 25 ppm to 510 ppm, 30 ppm to 510 ppm, 35 ppm to 510 ppm 510 ppm, 40 ppm to 510 ppm, 45 ppm to 510 ppm, 50 ppm to 510 ppm, 55 ppm to 510 ppm, 20 ppm to 505 ppm, 25 ppm to 505 ppm, 30 ppm to 505 ppm, 35 ppm to 505 ppm, 40 ppm to 505 ppm, 45 ppm to 505 ppm, 50 ppm to 505 ppm, 55 ppm to 505 ppm, 20 ppm to 500 ppm, 25 ppm to 500 ppm.30 ppm to 500 ppm, 35 ppm to 500 ppm, 40 ppm to 500 ppm, 45 ppm to 500 ppm, 50 ppm to 500 ppm, 55 ppm to 500 ppm, 20 ppm to 495 ppm, 25 ppm to 495 ppm, 30 ppm to 495 ppm, 35 ppm to 495 ppm, 40 ppm to 495 ppm, 45 ppm to 495 ppm, 50 ppm to 495 ppm, 55 ppm to 495 ppm, 20 ppm to 490 ppm, 25 ppm to 490 ppm ppm, 30 ppm to 490 ppm, 35 ppm to 490 ppm, 40 ppm to 490 ppm, 45 ppm to 490 ppm, 50 ppm to 490 ppm, 55 ppm to 490 ppm, 100 ppm to 400 ppm, 150 ppm to 400 ppm, 200 ppm to 400 ppm, 250 ppm to 400 ppm, 300 ppm to 400 ppm, 100 ppm to 150 ppm, 100 ppm to 200 ppm, 100 ppm to 250 ppm, or 100 ppm to 300 ppm. By maintaining the content within this range, it has the advantage of imparting a moderate sweetness to food and beverages. In this specification, unless otherwise specified, "ppm" refers to "ppm by mass".

[0135] The food and beverage products of the present invention may also contain sweeteners other than steviol glycosides. Examples of such sweeteners include fructose, sucrose, glucose-fructose syrup, glucose, maltose, sucrose, high-fructose syrup, sugar alcohols, oligosaccharides, honey, sugarcane juice (molasses), maltose, monk fruit powder, monk fruit extract, licorice powder, licorice extract, arrowroot seed powder, arrowroot seed extract, and other natural sweeteners, or artificial sweeteners such as acesulfame potassium, sucralose, neotame, aspartame, and saccharin. From the viewpoints of refreshingness, ease of drinking, natural taste, and imparting a moderate alcoholic aroma, natural sweeteners are preferred, and fructose, glucose, maltose, sucrose, and sucrose are particularly suitable. Only one of these sweeteners may be used, or multiple sweeteners may be used.

[0136] Regarding the content of sweeteners other than steviol glycosides, when the sweeteners are high-sweetness sweeteners (e.g., mogroside V, xylitol, and artificial sweeteners), the composition ratio of the steviol glycoside composition of the present invention to the sweeteners other than steviol glycosides, by weight, can be 1:99~99:1, 5:99~95:5, 10:90~90:10, 15:85~85:15, 20:80~80:20, 25:75~75:25, 30:70~70:30, 35:65~65:35, 40:60~60:40, 45:65~65:45, or 50:50. When the steviol glycoside composition of the present invention contains a low-sweetness sweetener (e.g., sucrose or high-fructose corn syrup), the composition ratio of the steviol glycoside composition of the present invention to the low-sweetness sweetener, by weight, may be 1:1000~1:100, 1:800~1:100, 1:700~1:100, 1:600~1:100, 1:500~1:100, 1:400~1:100, 1:300~1:100, or 1:200~1:100.

[0137] The food products of this invention are not particularly limited, and may include, for example, snacks, breads, cereal flours, noodles, rice, agricultural or forestry processed foods, livestock processed products, aquatic processed products, milk or dairy products, oils or oil-processed products, seasonings or other food raw materials.

[0138] The beverages used in this invention are not particularly limited, and examples include carbonated beverages, non-carbonated beverages, alcoholic beverages, non-alcoholic beverages, beer-flavored beverages such as beer or non-alcoholic beer, coffee beverages, tea beverages, cocoa beverages, nutritional beverages, functional beverages, etc.

[0139] The beverage of the present invention can also be prepared as a containerized beverage that has been heat-sterilized and packaged in a container. There are no particular limitations on the container; examples include PET bottles, aluminum cans, steel cans, cardboard boxes, chilled cups, and bottles. There are no particular limitations on the type of heat sterilization used; for example, conventional methods such as UHT sterilization and autoclave sterilization can be used. There are no particular limitations on the temperature of the heat sterilization process; for example, it is 65~130°C, preferably 85~120°C, for 10~40 minutes. Where sterilization effects equivalent to those described above can be obtained, sterilization can also be performed at an appropriate temperature for a few seconds, for example, 5~30 seconds.

[0140] The method for manufacturing the food and beverage of the present invention is not particularly limited as long as a food and beverage containing the above-mentioned ingredients can be obtained. According to one embodiment of the present invention, a manufacturing method is provided, which is a method for manufacturing the food and beverage of the present invention, characterized by comprising obtaining the steviol glycoside composition of the present invention and adding the steviol glycoside composition to the food and beverage or its raw materials. Obtaining the steviol glycoside composition of the present invention is as described in "1. Method for Manufacturing the Steviol Glycoside Composition" above. Adding the steviol glycoside composition of the present invention to the food and beverage or its raw materials can be carried out in any step of the food and beverage manufacturing process, for example, it can be carried out when mixing the raw materials of the food and beverage, or when finally adjusting the flavor of the food and beverage.

[0141] Example

[0142] The present invention will be described in detail below through examples, but the content of the present invention is not limited thereto.

[0143] <Manufacturing Example>

[0144] 1. Extraction and solid-liquid separation

[0145] Stevia leaves (moisture content: 3-4% by weight) were heated to 60°C ± 5°C in 15 times the volume of deionized water, and the dried stevia leaves were immersed in the water. Extraction was then performed for 60 minutes using a kneading extractor (SKN-R100, manufactured by Sanyu Machinery Co., Ltd.) with stirring at 8 rpm. The extract was then filtered through 18-mesh and 140-mesh sieves, cooled with cold water using a heat exchanger, and the filtrate was separated into solid and liquid components using a disc centrifuge (9150 rpm (11601G), 24 L / min) to obtain a primary extract. During this process, the filtered leaves were extracted again under the same conditions, and the solid and liquid components were separated to obtain a clear secondary extract, which was added to the primary extract to obtain a solution containing a steviol glycoside composition.

[0146] 2. Cohesion

[0147] A first treatment solution (calculated by Brix (concentration of soluble solids)) equivalent to 16.16% of the soluble solids in the steviol glycoside solution was added to the steviol glycoside solution, and the mixture was stirred for 15 minutes. Then, a second treatment solution (treatment solution) equivalent to 28.28% of the soluble solids in the steviol glycoside solution was added to the first treatment solution, and the mixture was stirred for 30 minutes. After adjusting the pH to 7 with citric acid to prepare the second treatment solution, a 0.5% (w / v) chitosan solution was further added at a volume (mL) equivalent to 5.63 times the soluble solids (g) in the first treatment solution. The mixture was vigorously stirred for 3 minutes, gently stirred for 2 minutes, and allowed to stand for 10 minutes. Then, the electrically neutral coagulated precipitate was removed by centrifugation. The second treatment solution was thus clarified.

[0148] 3. Resin purification

[0149] As a resin purification process, (i) purification using anion exchange resin and (ii) purification using hydrophobic porous resin (resin without introduced ion exchange groups) are implemented.

[0150] (i) Purification using anion exchange resin

[0151] A highly porous basic anion exchange resin (manufactured by Mitsubishi Chemical Corporation) was packed into a column, and the second treatment solution after coagulation and separation was added to the column for purification. After adding the second treatment solution after coagulation and separation into the column, it was then pressurized with twice the volume of ion-exchanged water to recover the purified solution containing the steviol glycoside composition. This purification process removes black impurities and coloring components from the treatment solution.

[0152] (ii) Purification using hydrophobic porous resins

[0153] A hydrophobic porous resin (manufactured by Mitsubishi Chemical Corporation) was packed into a column, and the purified sample (i) obtained using anion exchange resin was added to the column for purification. The hydrophobic porous resin used was a copolymer of styrene and divinylbenzene, lacking ion exchange groups, and having a modulopore radius of 45 Å. After adding the purified solution (i) to the column, the column was washed with 3 times the column volume of 0.01 M citric acid aqueous solution and 3 times the column volume of 0.01 M sodium hydroxide aqueous solution. Then, the steviol glycoside composition was eluted with 4 times the column volume of 60% ethanol aqueous solution and recovered.

[0154] 4. Evaporation and Concentration

[0155] The solution was concentrated and ethanol was removed simultaneously using the EVAPOR centrifugal thin-film vacuum evaporator (manufactured by Ōkawahara Seisakusho). After the evaporation and concentration process, water remained, and the steviol glycoside composition was in liquid form.

[0156] <Example 1>

[0157] To evaluate the pH dependence of steviol glycoside concentration in solutions containing steviol glycosides derived from stevia extract.

[0158] Except that the amount of ion-exchange water was set to 30 times that of dried stevia leaves (moisture content: 3-4% by weight), and the extraction was performed once, a solution containing steviol glycosides was prepared using the same method as in "1. Extraction and solid-liquid separation" of the <Manufacturing Example> above. At this time, the pH of the solution containing steviol glycosides was measured to be 5.7. Then, NaOH (manufactured by Nacalai Tesque Co., Ltd., 4 mol / L sodium hydroxide solution) or citric acid (manufactured by Fujifilm and Wako Pure Chemical Industries Co., Ltd., product number 038-06925, purity ≥ 99.5%) was added to the solution containing steviol glycosides to gradually change the pH from about 3 to about 12, and the concentrations of rebaudioside A, rebaudioside D, and rebaudioside M were measured at each pH.

[0159] Next, citric acid was added to the solution containing steviol glycosides, which was adjusted to a pH of approximately 12, until the pH was lowered to approximately 7, and the concentration of riboflavin A was measured again.

[0160] The concentration shifts of the three steviol glycosides mentioned above as the pH was gradually varied to approximately 12 are shown in the figure. Figure 1 The change in the content of riboside A when the pH was then lowered to approximately 7 is shown in the figure. Figure 2 In addition, the concentrations of each steviol glycoside were determined by liquid chromatography-mass analysis (LC / MSMS) under the conditions shown in Table 1 below.

[0161] [Table 1]

[0162]

[0163] like Figure 1 As shown, the concentration of rebaudioside A increases around pH 9, and then increases significantly at pH 12. On the other hand, no increase or decrease in the concentration of rebaudioside D and rebaudioside M was confirmed even when the pH was changed to alkaline.

[0164] This result confirms that the pH-dependent concentration of riboside A increases in solutions containing steviol glycosides.

[0165] In addition, such as Figure 2As shown, after the pH was raised to approximately 12, the concentration of ribobadiin A in the steviol glycoside-containing solution was not significantly altered when the pH was lowered to approximately 7 using citric acid. This result confirms that after the pH was raised to approximately 12, ribobadiin A remained stable in the steviol glycoside-containing solution even when the pH was lowered back to the neutral region.

[0166] <Example 2>

[0167] For Rebaudioside A, whose concentration increased in the solution containing steviol glycosides as confirmed in Example 1, it was evaluated whether the increase in concentration was due to the type of alkali.

[0168] A solution containing steviol glycosides was prepared using the same method as in Example 1 above. The pH of the solution containing steviol glycosides was then measured and found to be 5.8. Next, four samples were prepared with different types of alkali to make the pH alkaline. The pH values ​​of each sample after pH adjustment are shown in Table 2.

[0169] • Sample 1: No alkali agent (no change in pH)

[0170] • Sample 2: NaOH (manufactured by Nacalai Tesque Co., Ltd., 4 mol / L sodium hydroxide solution)

[0171] • Sample 3: KOH (manufactured by Nacalai Tesque Co., Ltd., 1 mol / L potassium hydroxide solution)

[0172] • Sample 4: Sodium bicarbonate (NaHCO3) (manufactured by Kanto Chemical Co., Ltd., product number 58024-17, purity ≥ 99.0%)

[0173] Then, for each of the pH-adjusted samples 1-4, the concentration of riboboroside A was determined using the same method as in Example 1. The results are shown in... Figure 3 .

[0174] [Table 2]

[0175]

[0176] like Figure 3 As shown, in Sample 2 (NaOH) and Sample 3 (KOH), where the pH was adjusted to approximately 12, the concentration of rebaudioside A was significantly increased compared to Sample 1, where the pH was not changed from 5.8. This result confirms that the increase in the concentration of rebaudioside A is pH-dependent, not dependent on the type of alkali.

[0177] <Example 3>

[0178] To investigate the manufacturing parameters in the industrial production of steviol glycosides, the changes in steviol glycoside concentration were evaluated by varying pH and holding time.

[0179] Solutions containing steviol glycosides were prepared using the same method as in Example 1 above. Next, the pH of the steviol glycoside-containing solutions was adjusted to 9, 10, 11, 11.8, 12, 12.2, and 12.5 using NaOH. These pH values ​​were maintained, and samples adjusted to each pH were kept at room temperature (approximately 25°C) for the time intervals shown in Table 3. Then, the pH of each sample was adjusted to approximately 7 using citric acid, and the concentration of steviol glycosides was determined using the same method as in Example 1. The increase rate of steviol glycoside concentration in samples adjusted to each pH value is shown in Tables 4-9, based on the sample at the moment the pH was adjusted to each specified value (0 minutes). Additionally, Table 4 shows riboflavin A, Table 5 shows riboflavin C, Table 6 shows steviol glycosides, Table 7 shows riboflavin F, Table 8 shows riboflavin G, and Table 9 shows the increase rate of concentration of raspberry glycosides.

[0180] [Table 3]

[0181]

[0182] [Table 4]

[0183]

[0184] [Table 5]

[0185]

[0186] [Table 6]

[0187]

[0188] [Table 7]

[0189]

[0190] [Table 8]

[0191]

[0192] [Table 9]

[0193]

[0194] As shown in Tables 4-9, it was confirmed that none of the steviol glycosides were pH-dependent or time-dependent, and the steviol glycosides remained stable in solution after increasing their concentration. Furthermore, a significant increase in concentration was observed in all of the above steviol glycosides in samples adjusted to pH 10-13.

Claims

1. A method for manufacturing a steviol glycoside composition, characterized in that, This includes preparing a solution containing steviol glycosides derived from stevia extract. Additives are added to the solution containing steviol glycosides to prepare the treatment solution. The additive is added to achieve a pH of 10-13 in the treated solution.

2. The manufacturing method according to claim 1, characterized in that, The additive is an alkali agent.

3. The manufacturing method according to claim 2, characterized in that, The alkaline agent contains one or more compounds selected from alkali metal hydroxides, alkaline earth metal hydroxides, silicates, carbonates, and bicarbonates.

4. The manufacturing method according to any one of claims 1 to 3, characterized in that, The steviol glycoside composition contains one or more steviol glycosides selected from ribobadiin A, ribobadiin B, ribobadiin M, ribobadiin C, ribobadiin D, steviol glycoside, ribobadiin F, dukorbitol A, ribobadiin G, ribobadiin N, raspberry glycoside, and steviol disaccharide glycoside.

5. The manufacturing method according to any one of claims 1 to 4, characterized in that, After preparing the treatment solution, keep it for 1 minute to 3 days.

6. The manufacturing method according to any one of claims 1 to 5, characterized in that, The process further includes adding a second additive to the treatment solution to prepare a second treatment solution. The second additive is added in such a way that the pH of the second treatment solution reaches 2 to 10.

7. The manufacturing method according to claim 6, characterized in that, Further, one or more compounds selected from aluminum sulfate, aluminum polychloride, ferric chloride (III) or its hydrates, polyacrylamide hydrolysate, alginic acid, chitosan and chitosan are added to the treatment solution and / or the second treatment solution.

8. A method for increasing the concentration of steviol glycosides in a solution containing steviol glycosides, characterized in that, This includes preparing a solution containing steviol glycosides derived from stevia extract. Additives are added to the solution containing steviol glycosides to prepare the treatment solution. The additive is added to achieve a pH of 10-13 in the treated solution.

9. The method according to claim 8, characterized in that, Increase the concentration of one or more steviol glycosides selected from ribobadiin A, ribobadiin C, steviol glycoside, ribobadiin F, ribobadiin G and raspberry glycoside.

10. A steviol glycoside composition, characterized in that, Manufactured by the method of any one of claims 1 to 7.

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