KR101924159B1 - Method for extracting soluble curcuminoid from turmericand soluble curcuminoid extracted by the same - Google Patents

Abstract

The present invention relates to a method for extracting an increased water-soluble cucuminoid using a steviol glycoside and to an increased water-soluble cucuminoid extracted therefrom.
The cucuminoids having increased water solubility extracted by the method of extracting cucuminoids with increased water solubility using the steviol glycoside of the present invention not only improve the solubility of the cucuminizoid, which is a poorly soluble material, in a water-soluble solvent, Cucuminizoid dissolved in a water-soluble solvent can be effectively used as a material for foods, medicinal products, and cosmetics since the original effect of cucuminoids, for example, antioxidative action and inhibition of blood glucose elevation can be sufficiently obtained.

Description

METHOD FOR EXTRACTING SOLUBLE CURCUMINOID FROM TURMERICAND SOLUBLE CURCUMINOID EXTRACTED BY THE SAME [0002] The present invention relates to a method for extracting a cucuminoid having increased water solubility using a steviol glycoside,

The present invention relates to a method for extracting an increased water-soluble cucuminoid using a steviol glycoside and to an increased water-soluble cucuminoid extracted therefrom. The cucuminoids having increased water solubility extracted by the method of extracting cucuminoids with increased water solubility using the steviol glycoside of the present invention not only improve the solubility of the cucuminizoid, which is a poorly soluble material, in a water-soluble solvent, Cucuminizoid dissolved in water can be used effectively as a material for foods, medicinal products and cosmetics since it can sufficiently obtain the effect of the original effect of the cucuminoids in aqueous solution, for example, antioxidative action and inhibition of blood glucose elevation .

Cucuminoids are yellow spices of polyphenol component extracted from the roots of Curcuma longa Linn (Zingiberaceae), a plant belonging to the ginger family of the Dong-dong Province, and aromatic methoxy phenol groups, α, β-unsaturated β-diketo linker, Due to its three main chemical functions including keto-enol tautomerism, it has multi-targeted biofunctions such as nervous system protection, anti-diabetic, anticancer, cardioprotection, and is a therapeutic and preventive agent for various chronic diseases. Active research and development are underway in the field of medicine as well as food.

However, curcumin is an insoluble substance having a solubility of less than 0.1 mg / ml, specifically 11 to 600 ng / ml, based on water. In general, since the substance may exhibit a physiological action in a dissolved state, It is closely related to the effect, which is an important property in the use of the substance. Therefore, solubilization is indispensable for weakly soluble substances such as curcumin.

Also typically increases kyumi solenoid when very unstable in alkaline conditions pH above 7.2, kept for about 30 minutes and decomposition over 90% (Stability of curcumin in buffer solutions and characterization of its degradation products. Journal of Pharmaceutical and Biomedical Analysis, 15, 1867 (Kinetics of alkaline degradation of the food pigments curcumin and curcuminoids. Journal of Food Science, 62 , 267-269), pH 7.4 for 10 minutes Studies have also shown that 90% of glucuronidase is degraded.

In addition, cucuminoids generally exhibit low intestinal permeability in the body. The apparent cellular permeability of cucuminoids is very low, about 3 × 10 ^-6 cm / s, which is known to be due to the fact that more than 10% of the cucuminoids are degraded by metabolism during cell permeation and more than 20% are accumulated in the cells.

For these reasons, bioavailability of cucuminoids is less than 1% in the case of oral ingestion, and various methods for increasing the bioavailability have been studied and suggested. Korean Patent No. 10-1258537 discloses a novel curcumin derivative that improves water solubility and stability, but it has a limited commercial efficiency because it undergoes chemical synthesis through various steps.

Curcuma longa L.) is a food material that has traditionally been safely used as a spice and natural coloring agent in the ginger root. It has been widely used in India and other Asian countries for the prevention of dementia, liver disease, depression, treatment of gastrointestinal diseases, relieving hangover, and preventing constipation. It is known that it has anti-inflammatory, anti-inflammatory, anti-inflammatory, antioxidant, antimicrobial and anti-cancer effects and various physiological activity effects such as cardiovascular diseases, metabolic diseases and autoimmune diseases. Therefore, various products such as capsules, ointments, energy drinks, It is utilized as material. In recent years, it has been attracting attention as a functional material known to be effective for Alzheimer's disease, brain damage protection, vascular dementia, diabetes and the like. The reason for this effect is due to the curcuminoid, a yellow coloring matter of Ulgum, and its major component is known as curcumin.

Curcuminoids are 2-6% in the roots of turmeric, and most of them are composed of curcumin, but about 17% of demethoxy curcumin and 3% of bisdimethoxy curcumin bisdemethoxy curcumin). At the hospital, the rootstock of curcuma longa is designated as turmeric, and the root of root is called turmeric. In other words, turmeric and turmeric are the roots of the same plant, but they are called turmeric, which is tied to the bottom of the turmeric and turmeric troughs.

On the other hand, steviol glycoside is a sweetener classified as a natural additive, and is produced by extracting from leaves of Chrysanthemum and Stevia. The sweetness is about 200 ~ 300 times of sugar. Stevia is native to Paraguay in South America, and locals have long been using dried leaves as sweeteners. It is currently used in low-calorie foods, carbonated drinks, confectionery, pickles, and the like. JECFA (Joint Food Additives Expert Committee), the result of safety evaluation, the allowable daily intake (ADI) is set at 0 ~ 4mg / kg, weight / day (steviol). The steviol glycosides include stevioside, rebaudioside, dulcoside, rubusoside, steviolside, and the like.

Precedent literature

1. Korean Patent No. 10-1258537

2. Korean Patent Publication No. 10-2015-64234

The inventors of the present invention discovered that when water extracts curcuminoids using steviol glycosides, the water solubility of the curcuminoids in the water-soluble solvent is remarkably improved and can be directly extracted, thus completing the present invention.

According to one embodiment,

(a) mixing a raw material containing curcumin and a steviol glycoside;

(b) dissolving the mixture in ethanol and stirring;

(c) centrifuging the mixture dissolved in ethanol;

(d) separating and removing the supernatant; And

(e) removing the ethanol contained in the separated supernatant to obtain a cucuminoid; A method of extracting an increased water-soluble cucuminoid using a steviol glycoside comprising

In the present invention, "dissolution" includes not only a state of being completely dissolved in water but also a state of being solubilized by micelles or the like, or a state of a liquid being visually dispersed uniformly in an aqueous solvent, As shown in FIG.

In the present invention, "increased water solubility" means that solubility in a water-soluble solvent such as distilled water is increased. In the present invention, "increased water solubility" refers to a complex of a cucuminoid and a steviol glycoside, and a complex of a cucuminoid and a steviol glycoside is prepared by mixing ethanol with a steviol glycoside and then centrifuging The extraction process does not form a clear covalent bond between the cucuminoids and the steviol glycosides, but it means a state in which a physically constant structure is formed by intermolecular bonding.

According to the method of extracting cucuminoids with increased water solubility using steviol glycosides according to the present invention, the content of cucuminoids extracted from the cucumbers or turmeric is 15% or more, 50% or more, 60% or more, 80% or more Or substantially entirely in a receiving state.

In the method of extracting an increased water-soluble glucuronidase using a steviol glycoside according to the present invention, the curcumin-containing raw material may be Ulugum or turmeric. At the hospital, the rootstock of curcuma longa is designated as turmeric, and the root of root is called turmeric. In other words, turmeric and turmeric are the roots of the same plant, but they are called turmeric, which is tied to the bottom of the turmeric and turmeric troughs.

In the method of extracting glucuronidase with increased water solubility using the steviol glycoside according to the present invention, it is possible to use all of the above-mentioned uroguanum or turmeric in a powder state or a raw material state as it is. The powder of urea or of turmeric is prepared by pulverizing raw coriander or turmeric, and the production method is not particularly limited.

In the method for extracting the increased water-soluble glucuronide using the steviol glycosides according to the present invention, it is preferable that the concentration of the wort silver dissolved in ethanol is 40% (w / v) or less. There is no significant difference in the amount of curcuminoids extracted when the concentration of ugum exceeds 40% (w / v).

In the method of extracting cucuminoids with increased water solubility using the steviol glycoside according to the present invention, the steviol glycoside is selected from the group consisting of stevioside, ribauidoside A, ribauidoside B, ribauidoside C , Ribauidoside D, ribauidoside E, rubusoidal, dulcoside A, steviolide, or a mixture thereof.

In the method for extracting the increased water-soluble glucuronidase using the steviol glycoside according to the present invention, the concentration of the steviol glycoside dissolved in ethanol may be 5-10% (w / v). Preferably, the concentration of the steviol glycoside may be 8% (w / v).

In the method for extracting increased water-soluble glucuronides using the steviol glycoside according to the present invention, the steviol glycosides may be purified or enzymatic reaction products. That is, in the present invention, it is possible to use the steviol glycoside itself, or to use all of the steviol glycoside enzyme reaction solution. According to one embodiment, it is possible that the enzyme is lactase.

In the method for extracting the increased water-soluble glucuronide using the steviol glycosides according to the present invention, the ethanol may be an 80 to 90% ethanol solution.

In the method of extracting water-soluble glucuronides with steviol glycosides according to the present invention, the step of dissolving the mixture in ethanol and stirring may be carried out for 10 minutes to 20 minutes. When the mixing time is less than 10 minutes, sufficient reaction for forming the complex does not occur. If the mixing time is more than 20 minutes, the reaction efficiency may be lowered due to unnecessary mixing process.

The present invention also provides a glucuronide extracted by the method of extracting the increased water-soluble glucuronidase using the steviol glycoside according to the present invention.

The cucuminizoid having increased water solubility extracted using the steviol glycoside according to the present invention may be a substance or a food additive which becomes an active ingredient of any one of medicines, veterinary medicines, quasi drugs, cosmetics and pesticides.

The increased water-soluble glucuronide extracted by the extraction method according to the present invention may have an antioxidant activity and / or an effect of inhibiting the increase of blood sugar.

The increased water-soluble curcuminoids extracted by the extraction method according to the present invention may have a property of inhibiting the activity of dengue virus.

The increased solubility of the water-soluble glucuronide by the extraction method according to the present invention improves the solubility in a water-soluble solvent, so that the effect that the original glucuronide did not exert can be sufficiently obtained. Further, the method of improving water solubility of the cucuminoids of the present invention is superior in terms of productivity and cost, has excellent safety of workers, and has high industrial value.

Figure 1 shows the results of measuring the solubility of the extracted glucuronides in one embodiment of the present invention.
Figure 2 shows thin film chromatographic results for an aqueous solution of the extracted glucuronidase in one embodiment of the present invention.
Figure 3 shows the extraction efficiency of water-soluble glucocinoids according to the amount of water in the ethanol solution in one embodiment of the present invention.
Figure 4 shows the extraction efficiency of the water-soluble glucuronide according to the concentration of the steviol glycoside added in one embodiment of the present invention.
Figures 5 and 6 show the extraction efficiencies of the water-soluble glucocinoids according to the concentration of urea added in one embodiment of the present invention.
Figures 7 and 8 show the results of measuring DPPH activity of the extracted glucuronidase in one embodiment of the present invention.
9 shows the results of measuring the size of the carcuminoid particles extracted according to an embodiment of the present invention.
FIG. 10 shows changes in blood glucose level after the administration of glucuronide extracted according to an embodiment of the present invention.
FIG. 11 shows the results of measurement of the degree of inhibition of dengue virus activity of the curcuminoids extracted according to one embodiment of the present invention.

Hereinafter, various embodiments are provided to facilitate understanding of the present invention. The following examples are provided to facilitate understanding of the invention and are not intended to limit the scope of the invention.

< Example  1> Using steviol glycosides Ulgum  extraction

300 mg of corn gluten powder was dissolved in 1 ml of ethanol to prepare a 30% corn ethanol solution, and 100 mg of stevioside and ribaudioside as steviol glycosides were dissolved in 1 ml of ethanol to prepare a steviol A glycoside ethanol solution was prepared.

The above solutions were mixed to prepare an Eppendorf tube, which was prepared by mixing stevioside and turmeric powder (Tum-Ste), and mixing rebaudioside and turmeric powder (Tum-RebA) Lt; / RTI &gt;

The stirred solution was centrifuged at 12,000 rpm for 10 minutes, and the clear solution of the upper layer was transferred to another Eppendorf tube, and the ethanol was evaporated to give a powder.

As a comparative example, Tum-Water and Tum-Ethanol were dissolved in distilled water alone (Tum-Water) and 100% ethanol (Tum-Ethanol) were prepared in the same manner. The clear solution of the upper layer was recovered to evaporate ethanol, The remainder was again dissolved in water (for Tum-Water) or ethanol (for Tum-Ethanol).

< Experimental Example  1> extracted Cucuminoid  Water solubility measurement

Ethanol was evaporated and the remaining ugly powder was dissolved in 1 ml of distilled water (for Tum-Ste, Tum-RebA and Tum-SG) as a water-soluble solvent and centrifuged at 12,000 rpm for 10 minutes. The clear supernatant was filtered through a 0.20 ㎛ membrane filter Agilent, Santa Clara, Calif., USA). The results are shown in FIG.

1 is as follows.

Ste: Stevioside aqueous solution

RebA: Rebaudioside A aqueous solution

SG: Steviol glycoside aqueous solution

Tume-Ste: A sample obtained by dissolving ugly powder with stevioside in ethanol, and then evaporating the ethanol to obtain an extract

Tum-RebA: A sample obtained by dissolving Ulymum powder together with Riboidoside A in ethanol, and then evaporating the ethanol to obtain an extract obtained by dissolving the extract in water

Tume-SG: A sample obtained by dissolving Ulyumus powder with ethanol with steviolglycoside and then evaporating the ethanol.

Tum-water: A sample obtained by extracting the ugly powder with ethanol and then evaporating the ethanol with water

Tum-Ethanol: A sample obtained by extracting the ugly powder with ethanol and then evaporating the ethanol with ethanol

As shown in FIG. 1, it was observed that water was hardly soluble in water, but soluble in ethanol (Tum-Ethanol), stevioside, rebaudioside, and steviol glycoside prepared and dissolved in water . In particular, as shown in FIG. 1, stevioside (Tum-Ste), rebaudioside (Tum-RebA) and steviol glycoside (Tum-SG) were mixed with ethanol , It was confirmed that the water-soluble solvent, water, was well dissolved in water.

< Experimental Example  2> Chromatographic measurement

Ethanol was evaporated and the remaining ugly powder was dissolved in 1 ml of distilled water (for Tum-Ste, Tum-RebA and Tum-SG) as a water-soluble solvent and centrifuged at 12,000 rpm for 10 minutes. The clear supernatant was filtered through a 0.20 ㎛ membrane filter Agilent, Santa Clara, Calif., USA), and the solvent was evaporated to obtain an increased solubility of curcumin.

Thin film chromatography (TLC) was performed to observe whether the curcumin obtained as described above had increased solubility in an aqueous solution. The TLC plate was re-dissolved in methanol at a ratio of 0.5 (w / v) N- (1-Naphthy) ethylenediamine dihydrochloride and 5% (w / v) at a UV 254 mm wavelength using acetonitrile / water 85:15 / v) sulfuric acid was dissolved in the prepared coloring reagent and treated at 90 ° C for 3 min.

As a result, as shown in FIG. 2, it was confirmed that the water-soluble substance was increased after the ethanol treatment, and when the steviol glycosides, ribauidoside A or stevioside were treated with ethanol, (Fig. 2). (Lane 1: Tum-Ethanol, Lane 2: Tum-Ste, Lane 3: Tum-Reba, Lane 4: Tum-SG)

< Example  2> Depending on solution concentration Cucuminoid  Comparison of extraction efficiency

The amount of glucuronide contained in the final obtained aqueous extract and the extraction yield according to the concentration of the ethanol solution put into the mixture of stevioside, rebaudioside and steviolglycoside, as in the case of Example 1, And the results are shown in Table 1. &lt; tb &gt; &lt; TABLE &gt;

Equation 1

The amount of water during the ripening process *
(%, v / v) Acceptable cacuminoids using stevioside
(mg / g turmeric extraction) Aqueous cocumidolide using rebaudioside
(mg / g turmeric extraction) Aqueous cucuminoids using steviol glycoside
(mg / g turmeric extraction) 0 33.4 ± 1.3 17.0 ± 1.2 18.1 ± 0.6 10 30.0 ± 2.5 26.8 ± 1.6 23.1 ± 0.9 20 29.8 ± 2.0 25.5 ± 1.0 23.0 ± 1.8 30 28.9 ± 1.7 24.3 ± 1.6 17.3 ± 0.8 40 25.1 ± 2.5 19.7 ± 0.6 16.2 ± 0.6 50 24.0 ± 1.4 18.1 ± 1.7 16.1 ± 0.8 60 19.2 ± 0.9 11.2 ± 1.2 14.2 ± 0.5 80 14.6 ± 0.7 13.5 ± 0.6 8.7 ± 0.2

*: Represents the amount of water in the ethanol solution added to the mixture of ugly powder and steviol glycoside (for example, 0% water used 100% ethanol and 80% water used 20% ethanol).

As a result, when extracting the glucuronide using stevioside, the extraction yield is increased as the water content is minimized. When the rubaidoside contains 10% water (that is, when 90% ethanol solution is used ), And steviol glycosides also contained 10 to 20% water (i.e., 80% to 90% ethanol solution was used).

Among the three extractive materials used for the extraction of ugum, stevioside showed the best yield (Fig. 3).

< Example  3> Depending on the concentration of steviol glycosides Cucuminoid  Comparison of extraction efficiency

The amount of cupricuminolide was calculated according to the mixing ratio of the steviol glycosides mixed in the extraction of the urogens in Example 1, and the results are shown in Table 2.

Concentration of steviol glycosides
(%, v / v) Acceptable cacuminoids using stevioside
(mg / g turmeric extraction) Aqueous cocumidolide using rebaudioside
(mg / g turmeric extraction) Aqueous cucuminoids using steviol glycoside
(mg / g turmeric extraction) 0.5 0.2 0.04 0.1 One 0.8 1.7 0.4 2 6.6 ± 0.4 6.2 ± 0.1 2.8 ± 0.1 3 14.0 ± 1.1 12.1 ± 0.4 6.4 ± 0.5 4 19.9 ± 1.8 17.0 ± 1.2 9.0 ± 0.2 5 28.1 ± 2.6 20.9 ± 0.7 12.5 ± 0.8 6 27.4 ± 1.7 24.5 ± 1.9 16.5 ± 1.9 8 34.5 ± 1.4 32.4 ± 0.2 21.1 ± 0.3 10 33.4 ± 1.3 17.0 ± 1.2 18.1 ± 0.6

As a result, it was confirmed that the stiboside, ribeodioxide and steviol glycoside both had the best water-soluble yield when used at the concentration of 8% under the same conditions (Fig. 4).

< Example  4> Ulgum  Depending on concentration Cucuminoid  Comparison of extraction efficiency

In Example 1, the amount of cucuminoids hydrated was calculated according to the concentration of cucumber dissolved in ethanol at the time of cucumber extraction, and the results are shown in Table 3.

Ugly concentration
(%, v / v) Use StevioSide
Aqueous cocuminoid Use Ribau dioside
Aqueous cocuminoid Using steviol glycosides
Aqueous cocuminoid mg / g turmeric extraction mg / ml mg / g turmeric extraction mg / ml mg / g turmeric extraction mg / ml One 69.3 ± 4.0 0.7 70.6 ± 0.6 0.7 73.1 ± 1.0 0.7 3 56.2 ± 1.3 1.7 65.1 ± 0.4 2.0 65.8 ± 1.0 2.0 5 51.2 ± 3.7 2.6 ± 0.2 60.3 ± 0.9 3.0 61.3 ± 0.5 3.1 7.5 50.4 ± 0.2 4.3 48.4 ± 0.4 3.6 50.7 ± 0.7 3.8 ± 0.1 12.5 48.7 ± 0.5 6.1 ± 0.1 48.3 ± 0.6 6.0 ± 0.1 38.1 ± 0.6 4.8 ± 0.1 15 47.0 ± 3.2 7.0 ± 0.6 47.5 ± 0.2 7.1 37.3 ± 1.3 5.7 ± 0.2 17.5 44.4 ± 1.3 7.8 ± 0.3 47.1 ± 1.4 8.2 ± 0.2 31.8 ± 0.6 5.6 ± 0.2 20 38.1 ± 0.8 7.6 ± 0.2 45.8 ± 0.9 9.2 ± 0.2 30.8 ± 1.2 6.2 ± 0.2 25 34.7 ± 0.1 8.7 36.2 ± 0.5 9.2 ± 0.1 25.2 ± 0.2 6.3 30 34.5 ± 1.4 9.6 ± 0.6 32.6 ± 0.5 9.7 ± 0.1 22.3 ± 0.3 6.7 ± 0.4 35 32.3 ± 2.6 11.3 ± 1.1 27.7 ± 0.1 9.7 17.0 ± 1.1 5.9 ± 0.4 40 24.8 ± 0.8 9.9 ± 0.4 12.6 ± 1.9 5.0 ± 0.7 11.4 ± 1.0 4.6 ± 0.4

When the other conditions were the same, the yield of hydrolysis was the best at 1% concentration of urogens added to ethanol for both stevioside, rebaudioside and steviol glycosides (Fig. 5).

On the other hand, the highest amount of curcuminoids was extracted at 30-35% concentration of cucuminoids added to ethanol, and when the concentration of cucurbitides added to ethanol was higher than 35% There was no significant difference in the extraction amount (FIG. 6).

< Experimental Example  3> extracted Cucuminoid  Antioxidant properties

DPPH radical scavenging activity (DPPH radical scavenging activity) experiments were conducted to confirm the physiological characteristics of the glucuronides extracted through the above examples.

The DPPH radical scavenging activity was measured according to methods known in the art. DPPH solution (45 占 퐂 / ml methanol) was mixed with the extract obtained in the above example, and the decrease in absorbance was measured at 515 nm for 30 minutes at intervals of 30 seconds.

The radical scavenging activity was calculated based on 100% reduction of the absorbance of vitamin C and the concentration of the sample when the absorbance of DPPH was reduced by 50% is indicated by IC ₅₀ , and is shown in Table 4 and FIG.

DPPH radical-scavanging activity (IC ₅₀ (/ / ml)) Curcumin Reagent (DMSO Treatment) 36.24 + - 2.55 Tum-Ste 33.25 + - 1.29 Tum-Reb 47.50 ± 2.18 Tum-SG 64.68 ± 2.59

As shown in Table 4, the cacuminoids generally showed almost no hydroxylation, whereas the water-soluble cacuuminoids extracted with steviosides, ribaudiosides and steviolglycosides showed antioxidative activities, As can be seen, it was confirmed that the glucoside extracted from stevioside exhibited similar or somewhat improved antioxidant properties to curcumin dissolved in DMSO.

< Experimental Example  4> water soluble Increased Cucuminoid  Antioxidant characterization analysis

The antioxidative activity of water extracts from water extracts from uleum powder was confirmed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging method. Each sample was dissolved in water to a final concentration of 10 ~ 500 ㎍ in 100 ㎛ (DPPH). After incubation at 25 ° C for 30 minutes, the concentration was determined at 517 nm using a microplate reader (Molecular Devices, Sunnyvale, CA, USA). Curcuminoids (≥ 94%), α-tocopherol, and trolox were used as positive standards, and the DPPH radical-scavenging activity was determined by the following equation 2, and the results are shown in FIG.

Equation 2

In Fig. 8, the SC ₅₀ values of the cucuminizoids hydrolyzed with Ste, RebA and SG were measured as 127.6 ± 4.9, 105.4 ± 1.8, and 109.8 ± 3.2 ㎍ / ㎖, respectively, and Curcuminoids (≥≥ 94 %), α-tocopherol, and SC ₅₀ value for trolox is a 36.2 ± 2.6 ㎍ / ㎖, 3.5 ± 0.1, and 8.4 ± measured in 0.1 ㎍ / ㎖ antioxidant activity of the water-soluble larger kyumi extracted by the present invention cannabinoids significantly , Respectively.

< Experimental Example  5> extracted water solubility Cucuminoid  Check pH stability

The stability of the aqueous soluble glucuronidase extracted from Example 1 according to pH was confirmed at pH 6 to 10. (Use buffer: 50 mM Na-P-pH 6.0, pH 7.0, pH 7.5; NaOH-glycin-pH 8.0 to pH 10.0). 2 μg of cucuminoids extracted from Example 1 were dissolved in a pH of 6 to 10, stored at 25 ° C. for 1 week, and centrifuged at 12,000 g for 10 minutes. The supernatant was filtered through a 0.20 μm membrane to obtain a water soluble cucuminoid The amount was confirmed. The amount of cucuminoids was determined by measuring the absorbance at 425 nm or by using the TLC method.

pH receptivity Cucuminoids  Remaining amount ( % ) Stevioside ( Ste ) Rebaudioside A  ( Reba ) Steviol glucoside ( SG ) 6.0 80.3 ± 0.4 81.6 ± 0.9 82.3 ± 2.4 7.0 87.2 ± 4.1 84.2 91.0 ± 0.9 7.5 85.3 ± 2.7 84.8 ± 0.6 93.6 ± 2.8 8.0 99.2 ± 0.4 84.1 ± 3.3 90.9 ± 0.6 10.0 100 99.1 ± 0.6 85.0 ± 0.6

The relative amounts in proportion to the remaining amount at pH 6.0 were 80.3 ± 0.4, 81.6 ± 0.9, and 82.3 ± 2.4%, respectively.

The water-soluble glucuronidase extracted by the present invention was maintained at 84% or more even when the pH was 7 or higher, and was stable at various pH values. In addition, it was confirmed that the water-soluble cucuminoids prepared in the present invention were stable in aqueous solutions having a wide pH.

< Experimental Example  6> Encapsulated Cucuminoid  Particle size analysis

To analyze the particle size of the encapsulated cucuminoid complex (complex with stevioside etc.), the stevioside, ribauidoside, steviol glucoside cucuminoid-stevioside complex (Tum-Ste ), Cucuminoid-rebaudioside complex (Tum-RebA), and cucuminoid-steviol glucoside complex (Tum-SG) were dissolved in 10 ml of water. Dynamic light scattering (DLS) was confirmed using a Zetasizer Nano ZS (Malvern Instruments Ltd, Malvern, UK) to determine the particle size of each material.

Fig. 9 shows the results of measuring the particle size of the urophyllum powder, which was hydrolyzed with Ste, RebA and SG, the urophyllum powder extracted with ethanol, and the steviosides, ribaudioside and steviol glucoside.

As shown in FIG. 9, the particle sizes of Ste, RebA and SG were 2.4 nm, 3.35 nm and 2.6 nm, respectively, and the particle sizes of extracts extracted with ethanol alone, Ste, RebA and SG were 2.0 nm and 110.8 nm, 95.7 nm, and 32.7 nm, the cupose size according to the present invention is considered to be bioavailable with a particle size of 30 to 120 nm and a particle size of nanosize.

< Experimental Example  7> Aqueous Cucuminoid  Blood sugar increase inhibition characteristic

Male Sprague-Dawley (SD) rats were purchased from a 6-week-old male rats and were adapted for 1 week in an animal laboratory maintained at a humidity of 50% and a temperature of 22 ± 1 ° C. After fasting for 16 hours before the experiment, After 1 g of maltose was injected intraperitoneally, 0.75 mg of Tum-Ste and 10 mg of stevioside were administered based on 1 kg of body weight. Whole blood collected from the tail vein was counted with a glucose meter. The control group was administered with water. The change in the measured blood glucose was confirmed and the results are shown in Fig.

Referring to FIG. 10, when maltose was administered and blood glucose level rapidly increased for 15 to 30 minutes, no significant change was observed when the steviol glycosides were administered, but when glucuronide (Tum-SG) Showed that the rise in blood sugar level was greatly eased and the blood glucose level was maintained constantly. It was confirmed that curcuminoids extracted with stevioside showed an excellent level of glucose uptake inhibition by water - soluble curcumin.

< Experimental Example  8> Aqueous Cucuminoid  Dengue virus ( DENV4 )of NS2B - NS3 ^pro  Activity inhibition characteristic

The recombinant DENV4 NS2B-NS3 ^pro enzyme was prepared ( in vitro evaluation of novel inhibitors against the NS2B-NS3 protease of dengue virus type 4. Molecules, 18 , 15600-15612) And the inhibitory activity against NS2B-NS3 ^pro was confirmed. To 100 μl of the final enzyme reactor was added 0.04 U of enzymes and 1.65 μl of a fluorescent tetrapeptide substrate [benzoyl-norleucine-lysine-arginine-7-amino-4-methyl coumarin (AMC) (Bachem, Bubendorf, Switzerland) The test material and 40 mM Tris buffer (pH 7.5) were added. The final activity of the remaining enzyme according to the inhibition samples was confirmed using a SpectraMax Gemini XPS apparatus (Molecular Devices, Sunnyvale, Calif., USA) (λex = 380 nm, λem = 460 nm). Curcuminoids (purity ≥ 94%) were used as positive controls.

The degree of inhibition was determined by the following formula 3.

Equation 3

Control after 20 minutes reaction of C - (enzyme, buffer, enzyme substrate mixture)

C _o - Fluorescence of the control at 0 hours,

After 20 min of S - (enzyme, test sample, enzyme substrate mixture), fluorescence,

S _o - Fluorescence before reaction of (enzyme, test sample, enzyme substrate mixture).

IC ₅₀ is the concentration of material that can lower the activity of NS2B-NS3 ^pro by 50% in the absence of inhibitor.

As shown in FIG. 11, inhibition of dengue virus against the NS2B-NS3 ^pro protease was confirmed by the use of Steer, RebA and SG, and the IC ₅₀ values were 14.1 ± 0.2 and 24.0 굇 0.4 , And 15.3 ± 0.4 ㎍ / ㎖, respectively.

Comparison was increased kyumi dissolved in DMSO used in Example solenoid control group is confirmed that the IC ₅₀ value represents a 5.2 ㎍ / ㎖ inhibitory activity for NS2B-NS3 ^pro protease of dengue virus of a receiving screen turmeric extract of the present invention to .

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (10)

(i) mixing a solution obtained by mixing ethanol with a curcumin-containing raw material and a solution containing ethanol mixed with a steviol glycoside;
(ii) dissolving the mixture in ethanol;
(iii) centrifuging;
(iv) separating and removing the supernatant;
(v) removing ethanol contained in the separated supernatant to obtain a powder; And
(vi) dissolving the obtained powder in distilled water and centrifuging to extract the curcuminoid from the supernatant; To a method for extracting glucuronidase having increased water solubility using steviol glycoside,
Wherein the steviol glycoside is selected from the group consisting of stevioside, ribauidoside A, ribauidoside B, ribauidoside C, ribauidoside D, ribauidoside E, rubusoide, dulcoside A, &Lt; / RTI &gt; and mixtures thereof. &Lt; RTI ID = 0.0 &gt;
Extraction of glucuronides with increased solubility using steviol glycosides.
The method according to claim 1,
The curcumin-containing raw material may be selected from the group consisting of &lt; RTI ID = 0.0 &gt;
Extraction of glucuronides with increased solubility using steviol glycosides.
delete The method according to claim 1,
When the mixture of the curcumin-containing raw material and the steviol glycoside is dissolved in ethanol, the concentration of the steviol glycosides is 5-10% (w / v)
Extraction of glucuronides with increased solubility using steviol glycosides.
The method according to claim 1,
When the mixture of the curcumin-containing raw material and the steviol glycoside is dissolved in ethanol, the steviol glycosides are purified or enzymatic reaction products
Extraction of glucuronides with increased solubility using steviol glycosides.
The method according to claim 1,
When the mixture of the curcumin-containing raw material and steviol glycosides is dissolved in ethanol, the concentration of the ethanol is 80 to 90%
Extraction of glucuronides with increased solubility using steviol glycosides.
delete The method according to any one of claims 1, 2, and 4 to 6,
Wherein the particle size of the glucoside is from 2 to 120 nm
Extraction of glucuronides with increased solubility using steviol glycosides.
The method according to any one of claims 1, 2, and 4 to 6,
The curcuminoid exhibits the blood glucose-raising inhibitory properties
Extraction of glucuronides with increased solubility using steviol glycosides.
The method according to any one of claims 1, 2, and 4 to 6,
The curcuminoids exhibit the inhibitory activity against Dengue fever virus
Extraction of glucuronides with increased solubility using steviol glycosides.

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