KR20070039409A - Methods for increasing hmf content in asparagi tuber, and the method for extraction of hmf - Google Patents

Abstract

The present invention relates to a method for enhancing the content of hydroxymethylfurfural (HMF) in the astronomical dong and to a method for purifying the HMF content enhanced by the method,

Heat treatment of astronomical dong to enhance the content of HMF in astronomical dong, method of purifying HMF using ethanol extraction, solubility difference and various chromatography in astronomical dong with enhanced HMF content, and carriers to extract cheon dong dong with enhanced HMF, Provided are a health supplement functional food obtained by mixing an excipient, a diluent and the like or a pharmaceutical composition for preventing and treating an adult disease.

According to the present invention, it is possible to extract HMF, which is an active compound such as nitric oxide (NO) production inhibition, tyrosinase inhibition, and red blood cell sickling inhibition, resulting in the prevention of adult diseases with improved health functional food composition and its therapeutic effect. Therapeutic pharmaceutical compositions can be provided.

Astronomical dong, Heating, Hydroxymethylfurfural, HMF, Increased content, Extraction, Purification

Description

 Method for increasing HMF content in astronomical dong and method for purifying HMF content enhanced by the above method {Methods for increasing HMF content in Asparagi Tuber and

1 is a flow chart illustrating a method of extracting HMF from the extract after enhancing the content of HMF as an active compound in astronomical dong as an embodiment of the present invention,

Figure 2 is an HPLC chromatogram measuring the change in the content of HMF in the astronomical dong before and after the heat treatment of the present invention,

Figure 3 is a graph showing the measurement of the change in HMF content for each temperature during the heat treatment of the present invention,

Figure 4 is a graph showing the measurement of the HMF content change with time during the heat treatment of the present invention, and

5 is a hydrogen nuclear magnetic resonance spectrum of HMF purified by one embodiment of the present invention.

The present invention relates to a method for enhancing the content of HMF in astronomical dong and to a method for purifying HMF enriched by the above method, and more particularly, to inhibit nitrogen monoxide (NO) production from heat treated astronomical dong extract, tyrosinase It relates to a method for enhancing the content of HMF as an active compound such as tyrosinase inhibition and sickling inhibition of red blood cells, and effectively extracting HMF as an active compound from the extract.

In general, Asparagi Tuber is a vine perennial herb belonging to the Liliaceae, with a length of 1 to 2 m, short root length, many fusiform bloated roots spread in all directions, and the main stem is vine-like. The branches are thin and smooth. 1 ~ 3 leaf-like branches are fresh, linear, pointed, 1 ~ 2 cm long, 1 ~ 1.2 mm wide, curved like a bow, and are glossy. HMF (5-hydroxymethyl furfural), which is known to be contained in astronomical dong, has been reported in the literature to show various physiological activities of inhibition of nitric oxide (NO) production, tyrosinase inhibition, and sickling inhibition of red blood cells.

The structural formula of HMF is as follows.

[constitutional formula]

As reported in the literature, literature on inhibition of nitric oxide (NO) production of HMF is described by "A nitric oxide synthesis inhibitor from the roots of" by Kim, N. Y., Kang, T. H., Kim, D. H.Gentiana scabra in RAW 264.7 Cells. "Korean J. Pharmacognosy (1999),30, 173-176, and the literature on tyrosinase inhibition is described by Sharma, V. K., Choi, J., Sharma, N., Choi, M., Seo, S. Y.In vitro anti-tryrosinase activity of 5- (hydroxymethyl) -2-furfural isolated fromDictyophora indusiata. "Phytother. Res. (2004)18, 841-844, and the literature on the sickling inhibitory effect of erythrocytes is described by Abdulmalik, O., Safo, MK, Chen, Q., Yang, J., Brugnara, C., Ohene-Frempong, K., Abraham, DJ, "5-hydroxymethyl-2-furfural modifies intracellular sickle haemoglobin and inhibits sickling of red blood cells." By Asakura, T. Br. J. Haematol. (2005)128, 552-561, and weak mutations are described by Proma, K., Mukhlesur, R. M. "Cytotoxicities of metabolites from aMonocillium species. "Pak. J. Pharmacol. (2002)19, 9-12.

Meanwhile, prior arts related to HMF include Korean Patent Laid-Open Publication Nos. 10-2001-0007251 (2001.02.14) and 10-2001-0007250 (2001.02.14), but the patents include apple juice and oxalic acid juice, respectively. In addition, the addition of sulfur in the soil was added to increase the amount of HMF, and the technology of increasing the amount of HMF against the heat treatment could not be found. No technology has been found in Korea for the astronomical dong.

Accordingly, an object of the present invention is to provide a method for increasing the content of HMF, the active compound in astronomical dong.

Another object of the present invention is to provide a method for purifying HMF from astronomical dongs treated to increase the HMF content.

Another object of the present invention is to obtain an HMF extract with such an increased content, thereby providing a functional food composition and pharmaceutical composition effective for the prevention and treatment of adult diseases such as inhibiting nitric oxide (NO) production, tyrosinase inhibition, and inhibiting sickling of red blood cells. Is to provide.

According to one aspect of the invention,

Provided is a method for enhancing the content of HMF in astronomical dong, characterized in that the astronomical dong is heated at 50-210 ° C. for 5 to 60 minutes.

According to the second aspect of the present invention,

Extracting the astronomical dong of which the content of HMF has been enhanced by the method of the first aspect with ethanol to obtain an extract;

Dispersing the extract in water, extracting the mixture with one solvent selected from the group consisting of dichloromethane and ethyl acetate, and concentrating to obtain an organic solvent soluble fraction; And

The organic solvent soluble fraction is concentrated and then silica gel, reverse phase silica gel, using a solvent selected from the group consisting of methanol, hexane, ethyl acetate, acetic acid, formic acid, chloroform, benzene and a solvent mixed with two or more selected from them. Purifying HMF using chromatography using a carrier selected from the group consisting of a high-speed liquid and Sephadex (Sephadex) is provided.

According to the third aspect of the present invention,

A health supplement functional food comprising HMF purified by the method of the second aspect, a carrier, an excipient, and a diluent is provided.

According to the fourth aspect of the present invention,

Provided is a pharmaceutical composition for preventing and treating adult diseases comprising HMF, carrier, excipient and diluent purified by the method of the second aspect.

Hereinafter, with reference to the accompanying Figure 1 with respect to the present invention will be described in detail.

HMF, the active compound in astronomical dong, maximizes its content by heating astronomical dong. The heating temperature is preferably heat-treated at 50-210 ° C. for 5-60 minutes, more preferably at 70-200 ° C. for 20-45 minutes, and most preferably at about 180-195 ° C. for about 55-65 minutes. It is not preferable that the content of HMF cannot be sufficiently increased as intended if it is out of the above range.

At this time, any kind of machine for heating can be used as long as the machine can properly heat the astronomical dong such as a heater, a pressurizer, as well as a self-made electric roasting machine.

Thus, the astronomical dong with increased HMF content is extracted with ethanol to obtain an ethanol extract. As an ethanol concentration, it is preferable to use 50-100%, preferably 95%, and the extracted extract is dried by distillation under reduced pressure or spray drying.

An appropriate amount, preferably about 3 times, of water is added to the dried product and dispersed therein, and then it is partitioned and extracted into an appropriate amount, preferably an equal amount of dichloromethane, chloroform, ethyl acetate, ether, ethanol, preferably dichloromethane. .

The obtained extract was dried by distillation, distillation under reduced pressure, or a spray dryer, and then separated by silica gel, reverse phase silica gel, Sephadex, preferably column chromatography using silica gel, etc. Fractions are obtained.

According to this method, a maximum of about 9.26 g of HMF can be obtained per kg of astronomical extract. The HMF content was analyzed to reach 13 times the pretreatment (0.71 ± 0.2 mg / g) per gram of ethanol extract.

In addition, the extraction method has the advantage of effectively extracting HMF, which is an active compound, while controlling nitric oxide (NO) production inhibition, tyrosinase inhibition, and red blood cell sickling inhibition from the extract.

The HMF thus obtained may be formulated according to known methods using commercially available ingredients such as excipients, carriers, diluents and the like which are acceptable in pharmaceutical, cosmetic or food composition.

The carrier, excipient and diluent herein include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl Pyrrolidone, cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil.

In addition to the above components, it may further include commercially available lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives and the like.

On the other hand, as an example of the formulation, the active ingredient may be mixed or diluted with the carrier, or may be contained in a capsule, sachet, paper or other kind of carrier. The carrier can then be a solid, semisolid, or liquid substance that acts as a vehicle, excipient or medium for the active ingredient to serve as a diluent. Thus, the resulting formulation may be in the form of tablets, pills, dispersants, granules, elixirs, suspensions, emulsifiers, solutions, syrups, aerosols, soft and hard gelatin capsules, sterile injectable solutions, sterile packaged powders and ointments, etc. It may be administered orally or parenterally.

HMF preparations obtained in this way can be released rapidly, continuously or, if necessary, with active ingredients after administration to a patient as a dietary supplement for health supplements or as a pharmaceutical composition for the prevention and treatment of adult diseases.

For example, the daily dosage of HMF as the active compound is usually within the range of about 1-20 mg per kg of body weight, and 100 mg to 10, in terms of water or ethanol extract when used as a dietary supplement. It is enough in the range of g.

Example

EMBODIMENT OF THE INVENTION Hereinafter, embodiment of this invention is described in detail with reference to drawings. However, the following examples are illustrative of the present invention, and the present invention is not limited thereto.

Example 1 Effect of Heat Treatment of Astronomical Copper on HMF Content

This embodiment will be described with reference to Figures 1, 2 and 5 with respect to the effect of HMF content by heating the astronomical copper.

First, as shown in the flow chart of Figure 1, the astronomical dong was collected to remove the foreign matter and then finely chopped or crushed, 3 kg of which was heated at 190 ℃ for 30 minutes using an electric roasting machine and then refluxed at 80 ℃ 95% ethanol After extraction, the mixture was filtered and the filtrate was concentrated under reduced pressure to obtain an extract of 194.5 g.

The extract thus obtained was dispersed in 1 L of distilled water, and then partitioned and extracted three times with the same amount of dichloromethane, followed by concentrating by mixing only the soluble portion in dichloromethane.

Then, silica gel column chromatography (Merck, Art No. 7734, column size: 3 × 39 cm, eluent: dichloromethane-methanol = 100-1 and gradient from 1-1 to the dichloromethane soluble fraction 14 fractions were obtained, and the third fraction was a fraction containing HMF. The fraction was reconstructed into silica gel column chromatography (Merck, Art No. 7734, column size: 1.2 × 29.5 cm). , Eluent: hexane-ethyl acetate = 20-1 starting with elution to 1-1 to elute) to give 16.7 mg of HMF with 97% purity on HPLC.

On the other hand, the HMF content obtained by repeating the same process except for the heat treatment was 0.71 ± 0.2 mg / g ethanol extract. Therefore, it can be seen that by performing the method of the present invention, the HMF content can be increased up to 9.26 ± 0.7 mg / g ethanol extract as compared with the conventional method.

Separately, the change in the HMF content in the astronomical dong ethanol for the astronomical ethanol extract and the astronomical ethanol extract that was not treated as a control was measured using an HPLC chromatogram and the results are shown in FIG. 2.

As can be seen in Figure 2, the upper control shows a very small HMF peak, while in the lower example it was confirmed that the peak is very increased.

In addition, the hydrogen nuclear magnetic resonance spectrum of the material of which the content of the astronomical copper extract was greatly increased was measured and the results are shown in FIG. 5.

As shown in FIG. 5, since the results of each nuclear magnetic resonance spectrum and the properties exhibited by HMF were exactly the same, this compound was identified as HMF.

Example 2- Confirming Optimum Heating Temperature of Astronomical Building

This embodiment will be described with reference to Figures 3 and 4 to confirm the optimum temperature during the heating treatment of the astronomical dong.

HMF content change according to heating temperature for the extract obtained in Example 1 was measured and the results are shown in FIG. 3. As shown in Figure 3, it was confirmed that the content of HMF is improved within the range of 50-210 ℃, in particular it was confirmed that the production of HMF is maximized at 190 ℃.

The change in HMF content according to heating time for the extract obtained in Example 1 was measured and the results are shown in FIG. 4. As shown in Figure 4, it was confirmed that the content of HMF is improved within the range of 5-60 minutes, in particular it was confirmed that the production of HMF is maximized at 60 minutes.

It is apparent to those skilled in the art that the present invention is not limited to the above-described embodiments, and that various modifications and changes can be made without departing from the spirit of the present invention.

That is, in the present invention, the astronomical dong as an example to illustrate the enhancement of the content of HMF, the same or similar effects can be obtained through the same heat treatment for similar plants containing HMF. Therefore, such modifications or variations are intended to fall within the claims of the present invention.

According to the present invention, the HMF content of active compounds, such as inhibition of nitric oxide (NO) production, inhibition of tyrosinase, and inhibition of sickling of red blood cells can be enhanced and extracted, resulting in improved health functional food compositions and therapeutic effects. The pharmaceutical composition for the prevention and treatment of an adult disease can be provided.

Claims (7)

Method for enhancing the content of HMF in astronomical dong, characterized in that the astronomical dong is heated for 5 to 60 minutes at 50-210 ℃ The method of claim 1, wherein the heat treatment is performed at 70-200 ° C. for 20-45 minutes. Extracting by extracting astronomical dong of which the content of HMF is enhanced by the method of claim 1 or 2 with ethanol; Dispersing the extract in water, extracting the mixture with one solvent selected from the group consisting of dichloromethane and ethyl acetate, and concentrating to obtain an organic solvent soluble fraction; And The organic solvent soluble fraction is concentrated and then silica gel, reverse phase silica gel, using a solvent selected from the group consisting of methanol, hexane, ethyl acetate, acetic acid, formic acid, chloroform, benzene and a solvent mixed with two or more selected from them. Purifying HMF using chromatography using a carrier selected from the group consisting of a high-speed liquid and Sephadex. Health supplement functional food comprising a carrier, excipient and diluent (100 mg to 10 g) in 100 mg to 10 g of HMF purified by the method of claim 3 The method of claim 4, wherein the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl Characterized in that pyrrolidone, cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil are used. Functional food for health supplement A pharmaceutical composition for preventing and treating adult diseases, comprising a carrier, an excipient, and a diluent (100 mg to 10 g) in 100 mg to 10 g of HMF purified by the method of claim 3. According to claim 6, The carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl Characterized in that pyrrolidone, cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil are used. Pharmaceutical composition for the prevention and treatment of adult diseases

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