KR101046349B1 - Garlic extract, its preparation method and storage method - Google Patents

Abstract

The present invention relates to a garlic extract prepared by extracting a sulfur compound as a main component of garlic in high purity and a method for producing the same. More particularly, the present invention relates to a garlic extract prepared by adding ethanol to crushed garlic, or by mixing and crushing garlic with ethanol, Producing; Aging the garlic mixture; Removing the garlic residue from the aged garlic mixture and obtaining a garlic extract; Removing the protein and polyphenol from the garlic extract; Extracting only the components having a molecular weight of 5,000 or less in the garlic extract from which the protein and the polyphenol have been removed; and a method for producing the garlic extract.

According to the present invention, by reacting garlic with ethanol at room temperature without heating or acid / base treatment, the active ingredient of the sulfur compound of garlic is preserved as it is, and the concentration of the ingredient is not changed with the lapse of time, Garlic extract without garlic can be obtained.

Garlic, extracts, sulfur compounds, proteins, polyphenols, frozen storage

Description

[0001] The present invention relates to a garlic extract, a method for preparing the same,

The present invention relates to a garlic extract prepared by extracting a sulfur compound as a main component of garlic in high purity and a method for producing the same. More particularly, the present invention relates to a garlic extract prepared by adding ethanol to crushed garlic, or by mixing and crushing garlic with ethanol, Producing; Aging the garlic mixture; Removing the garlic residue from the aged garlic mixture and obtaining a garlic extract; Removing the protein and polyphenol from the garlic extract; Extracting only the components having a molecular weight of 5,000 or less in the garlic extract from which the protein and the polyphenol have been removed; and a method for producing the garlic extract.

Garlic (allium sativum L.) is a perennial bulbous plant belonging to the genus Allium (liliaceae) allium, planted taxonomically together with onion, onion, leek and algae. It is one of the first cultivated plants It is one. The primary source of garlic is Central Asia, and the second origin is the Mediterranean region. Especially, according to the Egyptian short record, there are 22 ways of using garlic, including the treatment of heart disease, headache, tumors, and the eradication of parasites, so that garlic is not only used for general food, Prevention, and treatment of the disease.

In Korea, the record of mountain (蒜) is conveyed to Dangun myth of Danggun-yusa in Samguk Yusa, and it indicates that the mountain (蒜) refers to garlic and its cultivation history is very long in our country.

This garlic contains about 60% water, about 28% carbohydrate (mainly fructan-fructan polymer), about 2.3% organic sulfur compounds, about 2% protein (mainly allinase) (About 0.08%), a small amount of saponin (about 0.07%), and a small amount of sitosterol (about 0.07%), as well as organic amino acids (mainly arginine, arginine) , About 0.0015%), and other vitamins such as vitamin A, B1, B2 and C and inorganic substances such as calcium, potassium, copper, zinc, germanium, iron, magnesium, manganese, phosphorus, Is known to be contained in trace amounts.

In particular, garlic contains a relatively large amount of organic sulfur compounds as compared to other plants. Such organic sulfur compounds have the effect of preventing or treating various diseases, wherein the organic sulfur compounds include sulfides, disulfides, trisulfides, ) -Containing compound, which contains alliin (1%), methine (0.12%), isoalignan (0.06%) and cycloaliphatic (0.1%), L- glutamyl- - Cysteine also contains glutamyl-S-trans-1-propenyl cysteine (0.6%) and glutamyl-S-allyl-L-cysteine (0.4%).

In addition, when garlic is scratched or crushed, an enzyme called alliinase flows out. By the action of the enzyme, the alliin changes to allicin, , Which is known as the main component of the enzyme, is very unstable at room temperature, so that the content of allysine is reduced to half in 2 to 16 hours and is converted into other sulfur compounds (sulfide, disulfide, trisulfide and diethyl group) A unique complex incense is generated.

However, when sulfur compounds such as allysine and garlic are exposed to strong acids or high temperatures, their components are destroyed. In case of contact with air or exposure to light, they may disappear, and the protein or polyphenol The sulfur compound may be combined with the sulfur compound to rapidly degrade the compound. Therefore, there has been a serious problem in preserving these components of garlic for a long period of time while preserving them.

In addition, in the case of garlic injections and medicines using garlic, there is a problem in that there is a great risk of inducing toxins in the body by using an organic solvent or protein disintegration to remove heterologous proteins that may cause side effects to the human body.

In addition, in the past, the method of extracting sulfur compounds and allysine from garlic by using high temperature and high pressure or aging with strong acid has depended on the method of extracting sulfur compounds or allysine.

DISCLOSURE Technical Problem Accordingly, the present invention has been made in order to solve the problems of the prior art as described above, and it is an object of the present invention to provide a method and apparatus for preventing garlic from being damaged due to the fact that alicin and other sulfur compounds showing irritating and spicy taste, , And to provide a garlic extract which can maximize the therapeutic effect and a method for producing the same.

It is still another object of the present invention to provide a method for storing garlic extract so that the components of the sulfur compound, which is a main component of garlic, are not reduced or denatured despite the lapse of time.

In order to achieve the above object, the present invention provides a method for producing a garlic mixture, comprising the steps of: adding garlic to crushed garlic; or mixing and crushing garlic and ethanol; Aging the garlic mixture; Removing the garlic residue from the aged garlic mixture and obtaining a garlic extract; Removing the protein and polyphenol from the garlic extract; And extracting only a component having a molecular weight of 5,000 or less in the garlic extract from which the protein and the polyphenol are removed. The present invention also provides a method for producing a medicament using garlic.

Here, the method for producing a medicament using garlic may further comprise lyophilizing the garlic extract having a molecular weight of 5,000 or less and pulverizing the garlic extract.

The step of removing the protein and the polyphenol from the garlic extract may be performed by a primary filtration using a filter having a pore size of 10 to 30 탆, filtering the filtered garlic extract again with a filter having a pore size of 0.1 to 0.5 탆 It is preferable to carry out secondary filtration.

In order to achieve the above-mentioned object, the present invention provides a pharmaceutical product prepared by the above method and using garlic in which protein and polyphenol are removed.

According to another aspect of the present invention, there is provided a method of storing a medicament using garlic, the method comprising: storing garlic-based medicines manufactured according to the present invention at a temperature of 4 ° C or lower in a shaded space;

As described above, according to the configuration of the present invention, the following effects can be expected.

First, the present invention utilizes a method in which garlic is treated with heat or an acid / base treatment or with ethanol at room temperature without fermentation under a high temperature and high pressure environment, whereby the active ingredient of the sulfur compound of garlic is preserved as it is, It is expected that the concentration of the component does not change, so that there is no risk of deterioration.

Secondly, it is expected that the action of keeping the purity of the sulfur compound high by pulverizing the pharmaceutical product using the garlic produced by the present invention by the freeze-drying method.

Third, the garlic-based medicines manufactured by the present invention are stored in a shaded space by refrigerating, so that the purity of the sulfur compounds is not changed and can be used stably, and the concentration of sulfur compounds can be controlled The effect is expected.

Fourth, since the garlic-based medicines according to the present invention do not use harmful organic solvents or substances capable of harming the human body such as release of proteins, they can be safely administered to the human body. It is expected.

Fifth, when the plant protein is removed from garlic, it is expected to minimize the adverse effects caused by the heterologous protein.

Sixth, garlic sulfur compounds are relatively cheaply manufactured and used for the treatment of various cancers, thereby providing economical advantages compared to existing expensive cancer drugs. Moreover, by using natural garlic sulfur compounds, anticancer drugs manufactured by conventional artificial chemicals can be obtained It is anticipated that a number of side effects that may occur when used are minimized.

Hereinafter, the present invention will be described in more detail with reference to preferred embodiments.

The garlic extract according to the present invention, the method for producing and storing the garlic extract, and the method for preparing and storing the garlic extract according to the present invention are characterized in that a sulfur compound is extracted from garlic, and the extract has an environmentally- It is intended to be used for cosmetics, foods or medicines, particularly for injections, and has an anticancer effect so as to broaden the use area as a therapeutic agent for cancer diseases. It also induces allergy to human body, The plant protein which is a cause of denaturation and the polyphenol component which is a cause of denaturation are removed and then lyophilized to store the powder in a refrigerated state so that the garlic extract can maintain its efficacy even when stored for a long period of time.

The garlic extract prepared according to the present invention may be administered to the body by various methods such as invasive methods. In order to use the garlic extract in this way, first, the garlic extract must be aseptic. Secondly, (4) the osmotic pressure of the extract should be as high as possible, and (5) the pH of the extract should be as low as possible. pH should be close to the sixth. If the extract is administered to the body, there should be no tissue disorder.

The method for extracting the sulfur compounds of garlic according to the present invention for this purpose can be described as one embodiment.

First, 1 kg of garlic is crushed and 1 liter of ethanol having a concentration of about 40% is added, or a mixture of ethanol and garlic is formed by mixing undiluted garlic with ethanol and disintegrating.

Then, the garlic mixture is reacted at room temperature of 25 ° C to 30 ° C for 1 to 2 hours for aging. Garlic residue is removed therefrom, and the garlic extract is extracted by filtration using a diatomaceous earth filter.

Next, the garlic extract is stored in a freezer at about -20 DEG C for 3 to 4 hours to condense the protein, and then thawed at room temperature for 60 to 90 minutes to deposit the suspension.

Thereafter, the precipitate is removed, and the supernatant is obtained and centrifuged at about -15 ° C for about 30 minutes at about 8000 rpm to remove the suspended matter remaining in the extract.

Next, the precipitate was once again removed from the centrifuged extract, and a polycarp (polycler, protein and polyphenol adsorbent) having a concentration of about 2000 ppm was added to the supernatant and shaken for about 30 minutes, Proteins, and polyphenols.

Then, in order to remove the polyclonal complex from the extract containing the polyclonal complex, the mixture was filtered using a filter having a pore size of about 10 to 30 탆, and then filtered using a filter having a pore size of 0.1 to 0.5 탆. Filtration again filters out the remaining suspended solids.

Next, the thus-filtered extract is separated into an extract composed of a substance having a molecular weight of 5,000 or less using a PES membrane.

Thereafter, the thus obtained extract is pulverized by a freeze-drying method and stored in a shade-protected environment at a temperature of 4 DEG C or less.

In addition, the powder culture was examined for the use of powder as an injection.

It is noted that the above-described method and storage method of garlic extract according to the present invention are merely one embodiment, and it is possible to omit a part of the method or to carry out the method with different numerical ranges.

One embodiment of the method for producing garlic extract according to the present invention will be described in more detail as follows.

The above-described step of crushing garlic is for extracting sulfur compounds contained in garlic into a high-purity solution, and there is no limitation on the crushing method such as shredding or shredding garlic. However, alliin, which is a characteristic irritant odor and taste of garlic, is decomposed into allicin and pyruvic acid by allinase, and allicin is again decomposed into diallyl disulfide diallyl disulfide. Therefore, it is preferable to finely crush the garlic to obtain a sulfur compound having a high purity as high as possible. In an embodiment of the present invention, the fresh-cut garlic which has been removed is shredded with a mixer and is preferably made into garlic juice.

Then, the addition of ethanol having a concentration of about 40% to the crushed garlic causes the sulfur compounds contained in the garlic to dissolve well in the solution and to remove a large amount of the high-purity sulfur compound solution while removing the protein contained in the garlic . In this case, when a high concentration of ethanol is used (for example, 80% ethanol), a cryogenic preliminary freezing process for freezing the ethanol solution is required during the lyophilization process, and bubbles are generated in the garlic mixture even at low temperatures during the drying process, The use of an excessively high concentration of ethanol is undesirable in terms of production cost and production process.

For reference, methods for removing proteins include heat treatment or freezing, the use of protein degradation solutions such as 0.5 M EDTA, and protease enzymes such as proteinase K (1 mg / dl) And precipitation method. However, in the case of sulfur compounds of garlic, the strong acid treatment or the heat removal method of the protein causes the destruction of the garlic sulfur compounds, and the method using the proteolytic enzyme is a method in which when the proteolytic enzyme remains in the extract, It has a detrimental effect. Therefore, ethanol was used to remove proteins contained in garlic without affecting the sulfur compounds of garlic.

Here, the garlic is crushed and mixed with ethanol, but the garlic and ethanol are mixed and crushed.

Thereafter, the garlic mixture is allowed to react at room temperature of 25 ° C to 30 ° C. The garlic extract thus obtained is a yellow to reddish-colored oily liquid, and has a stimulating flavor original to garlic and a pungent taste inherent to garlic. Such taste and aroma are attributed to the major components of garlic, namely, allysine and sulfur compounds. Alicin is destroyed when heat is applied weakly, and the taste and incense of the above-mentioned garlic are lost. Conventional methods of processing garlic have been to ferment garlic and then extract the active ingredient of garlic in a manner that it must be heated, or to extract the active ingredient of garlic at high temperature and high pressure. According to this conventional method, Considering that the flavor and aroma are destroyed, the present invention necessarily causes the garlic mixture to react at room temperature, and this process is characteristic of the present invention.

In particular, since allysine in garlic is produced by the action of allysine enzyme and is highly unstable, the produced allysine and the sulfide compound, which is a metabolite thereof, rapidly react with time and become browned Since the properties change, it is necessary to react for a suitable time at room temperature.

According to this embodiment, the garlic mixture is stored at room temperature of 25 ° C to 30 ° C for about 1 to 2 hours, and allyl and allylase, which are generated at the time of pulverization of garlic, are reacted to produce allysine. The aging time of the garlic mixture is preferably 1 to 2 hours as much as possible, and the amount of the sulfur compounds actually extracted is reduced due to the inhibition effect on the alnase when the reaction time is longer or the reaction temperature is higher at room temperature.

Next, the residue is removed from the extracted garlic extract and filtered to obtain more purified garlic extract. More specifically, the aged garlic extract is firstly filtered using a mesh to be juiced. Subsequently, the juice solution is subjected to secondary filtration using a filter using diatomaceous earth to produce a more purified garlic extract. The diatomaceous earth filter used in this case is preferably a diatomaceous earth filter because it filters out all the particles having a particle size of 3 μm or more in the solution and also does not affect the components of the garlic extract, especially the sulfur compounds. Thus, the garlic extract, which has been subjected to diatomite filtration, has pale yellow and slightly turbid properties.

Then, the purified garlic extract is stored in a freezer at about -20 캜 for 3 to 4 hours to condense the protein, and then thawed at room temperature for 60 to 90 minutes to precipitate the suspension.

Then, the garlic extract precipitated with the suspension is centrifuged again at about -20 DEG C at a rotation speed of about 8,000 rpm for about 30 minutes to remove the remaining scum from the garlic extract, and then a transparent supernatant is obtained. When stored at room temperature or in a refrigerator, the supernatant becomes cloudy or precipitates after 24 to 72 hours.

This is due to the residual protein in the garlic extract and the polyphenol component of garlic. To remove it, the polyclor is mixed with the solution at a concentration of about 2,000 ppm, shaken for about 30 minutes, Is used to remove the protein and the polyclonal complex bound to the polyphenol.

The filtered garlic extract is filtered using a filter having a pore size of 0.2 mu m to obtain a transparent garlic extract. Garlic extracts filtered on a 0.2 μm filter are kept in a transparent state during refrigerated storage, but after 48-72 hours at room temperature, the solution becomes turbid.

Such turbid components are caused by a polymer component having a molecular weight exceeding 5,000 remained in the garlic extract. Such aggregation of the polymer component causes turbidity phenomenon. Therefore, in order to remove it, The extract is filtered using Vivaflow 50, a polyethylenesulfone (PES) membrane, and a filter solution having a molecular weight of 5,000 or less is obtained again.

The thus filtered garlic extract retained its transparency for a long time at room temperature and did not cause turbidity and showed negative in the biurette reaction. This filtering process enhances the purity of the sulfur compounds of the extract. After filtering, the purity of the sulfur compounds in the garlic extract is about 80% ~ 85% (about 15% ethanol).

The finally obtained garlic extract is dried in a freeze dryer at a temperature of -40 DEG C and 5 mTorr for use in cosmetics, food, and other medicines, while maintaining the purity thereof.

At this time, sublimation heat is not applied to the freeze dryer for sublimation of ethanol. This is because the sublimation hyphae applied during freeze drying may cause bubbles of garlic extract during the drying process or may affect the alicin and sulfur compounds in the garlic extract to reduce the concentration of sulfur compounds in the dry powder.

The dried powder obtained through such a drying process has a white or pale off-white color, and its taste has a sparkling, sparkling, but incense-free characteristic.

In addition, the major component of garlic, alicin and its metabolic sulfur compounds, is very unstable. Therefore, it is difficult to maintain its purity at room temperature or in a liquid state. In one embodiment of the present invention, the garlic extract is lyophilized to powder, It should be stored in a light-shielding environment, for example, a brown bottle capable of blocking light, and kept in a refrigerated state. Such a storage method is a feature of the present invention.

In the embodiment of the present invention, in order to maintain the purity of unstable sulfur compounds for a long time, it should be stored at a temperature of 4 ° C or lower, preferably -2 ° C to 4 ° C. When stored in this manner, it can be seen that the concentration of the sulfur compounds contained in the dry powder remains constant despite the passage of time.

When the extracted garlic extract is administered to cancer cells, the growth of cancer cells is inhibited. This will be described later.

≪ Experimental Example 1 > Relation between the concentration of ethanol and the amount of supernatant

In order to investigate the concentration of ethanol showing the maximum precipitation effect on protein removal in the process of producing garlic extract according to the present invention, solutions of 0%, 10%, 20%, 40% and 80% ethanol were used. Was stored at -20 to -40 < [deg.] ≫ C in a frozen state to observe the sedimentation rate over time.

As a result, the amount of the supernatant to the garlic extract of each solution is as shown in Table 1 below. At this time, the effect of protein removal was compared using 0.5 M EDTA solution in ethanol.

Here, EDTA is essential for maintaining the entire structure of the cell envelope and has the effect of inhibiting intracellular enzymes capable of removing magnesium (Mg) ions and cleaving DNA. EDTA at a concentration of 0.5M at a pH of 8.0 (500 ppm maximum) can also be used as a protein removal solution.

[ Table 1 ] Amount of supernatant according to freezing time

Frozen time (hour) One 2 3 4 5 6 12 24 48 72 96 Sample 1 Amount of supernatant 0 0 0 8.2 7.6 7.6 15.6 16.4 24.1 24.1 48 Turbid turbidity 0 0 0 0 0 0 2 2 3 3 3 Sample 2 Amount of supernatant 0 27.6 34.5 34.5 31.3 31.3 37.5 40.6 41.2 34.5 68 Turbid turbidity 0 2 2 2 One One 2 2 3 3 3 Sample 3 Amount of supernatant 34.5 36.2 37.9 34.2 12.9 19.4 f f f Turbid turbidity 0 One One One 0 One One f f f Sample 4 Amount of supernatant 0 0 0 0 f f f f f f f Turbid turbidity 0 0 0 0 f f f f f f f Sample 5 Amount of supernatant 0 0 f f f f f f f f f Turbid turbidity 0 0 f f f f f f f f f Sample 6 Amount of supernatant 6.7 6.7 6.9 11.7 12.7 4.8 31.3 54.1 55.2 64 Turbid turbidity 0 0 0 One One One 3 3 3 3 Sample 7 Amount of supernatant 9.7 32.3 32.3 29.4 47.8 50 56.8 56.9 84 Turbid turbidity One One 1-2 2 One 2 2 3 3 Sample 8 Amount of supernatant 45.5 45.5 18.2 16.7 6.7 25.8 f f f Turbid turbidity 2 2 One One One One f f f Sample 9 Amount of supernatant 0 0 0 0 f f f f f f f Turbid turbidity 0 0 0 0 f f f f f f f Sample 10 Amount of supernatant 0 0 f f f f f f f f f Turbid turbidity 0 0 f f f f f f f f f

Sample 1: A solution of 500 ppm of EDTA in 80% ethanol

Sample 2: a solution of 500 ppm of EDTA in 40% ethanol

Sample 3: A solution of 500 ppm of EDTA in 20% ethanol

Sample 4: A solution of 500 ppm of EDTA in 10% ethanol

Sample 5: Solution containing only 500 ppm of EDTA

Sample 6: Solution using only 80% ethanol

Sample 7: Solution using only 40% ethanol

Sample 8: Solution with only 20% ethanol

Sample 9: Solution with only 10% ethanol

Sample 10: Solution without ethanol

Turbidity: 0 (turbidity), 1 (slightly cloudy), 2 (slightly transparent), 4 (transparent)

f: freezing

As shown in Table 1 above, the amount of supernatant after 24 hours of freezing was in the order of Sample 7> 2> 3> 8> 6> 1 and the transparency of the supernatant was 6> 1> 7> 2> 3, 8 was the order. Also, although not shown, the boundary between the supernatant and the sediment was clearly observed in the order of samples 1,6> 2,7> 3,8.

Compared with the addition of EDTA and the turbidity of the supernatant depending on the ethanol concentration, the EDTA did not affect the turbidity, and the amount of the supernatant decreased when EDTA was added. Able to know. The boundary between the supernatant and the precipitate was more clear when EDTA was added. That is, the highest amount of supernatant can be obtained when only 40% ethanol is used without using a protein solubilizing agent such as EDTA.

<Experimental Example 2> Change of supernatant of frozen garlic extract after diatomite filtration according to thawing time

According to one embodiment of the present invention, the garlic extract filtered with diatomite was stored in a freezer at about -20 DEG C for 3 to 4 hours to condense the proteins, and then allowed to stand for 60 to 90 minutes while thawing at room temperature to precipitate suspensions. Changes in the supernatant over time are shown in Table 2 below. Here, each sample was the same as each sample in Table 1.

[ Table 2 ] Changes in supernatant with thawing time

Thawing time (min) 0 10 20 30 40 50 60 120 Sample 1 Amount of supernatant 0 0 2 1.6 2.1 3.1 3.9 Turbid turbidity 0 0 One 2 2 2 3 Sample 2 Amount of supernatant 27.58 40 38 33.3 34.3 38.7 44 Turbid turbidity One 2 2 2 3 3 3 Sample 3 Amount of supernatant 34.5 30 30 22.2 22.2 23.3 11.1 Turbid turbidity One One One One One One 2 Sample 4 Amount of supernatant 0/58 0/50 0/50 21.4 26.7 28.1 19.4 Turbid turbidity 0 0 0 0 0 One One Sample 5 Amount of supernatant 0/58 0/50 0/50 29 33.3 34.4 35.3 Turbid turbidity 0 0 0 0 0 0 One Sample 6 Amount of supernatant 6.7 6 6 5.3 7.4 7.7 8 Turbid turbidity One One 2 3 3 3 3 Sample 7 Amount of supernatant 10 30 38 34.1 41.2 41.2 35.7 Turbid turbidity One 2 2 2 2 2-3 3 Sample 8 Amount of supernatant 45.5 22.2 22.2 16.7 18.5 20.7 15.4 Turbid turbidity One One One One One One One Sample 9 Amount of supernatant 0/55 10 16 19.2 25 25.8 26.7 Turbid turbidity 0 One One 0 0 One One Sample 10 Amount of supernatant 0/55 0/50 0/50 3.9 4.2 9.5 10 Turbid turbidity 0 0 0 0 0 0 One

As shown in Table 2, the amount of supernatant after thawing was in the order of 7> 2> 4> 8> 3> 6> 1, and the transparency of the supernatant was 1,6 = 2,7> 3,8 .

Thus, it can be seen that the transparency is relatively high and the concentration of the ethanol solvent, which can obtain the supernatant most, is 40%.

The clear supernatant obtained above was subjected to protein qualitative analysis using the buret reaction and the protein was detected.

After cryopreservation and thawing, the supernatant was centrifuged at -15 ° C and 8000 rpm for 30 minutes to remove precipitates. A transparent supernatant was obtained, and the supernatant was mixed with the supernatant at a concentration of 2,000 ppm and shaken for 30 minutes. The melted protein and polyphenol were bound to the polyclonal and the polyclonal complex bound to the protein and polyphenol was removed using a 20 탆 filter as described above. Although the supernatant thus obtained was transparent, when the protein test (buret reaction) was performed, it was found to be slightly positive, indicating that a small amount of protein was present.

After removing the polycar, the supernatant was filtered again with a 0.2 μm filter, and then filtered using Viva Flow 50, a PES membrane, to obtain a garlic extract having a molecular weight of 5,000 or less. Thus, the protein test (buret reaction) of the garlic extract having a molecular weight of 5,000 or less resulted in a negative result.

<Experimental Example 3> Determination of concentration of sulfur compounds according to storage conditions of garlic extract

In order to investigate the influence of temperature on the components of sulfur compounds of garlic and the effect of the extract on the properties of garlic extracts prepared for long - term storage, two types of garlic extracts were prepared as liquid and freeze - And stored at -20 ° C for 3 days. The concentrations of sulfur compounds were measured at intervals of one month, and the results are shown in Table 3 below.

[ Table 3 ] Concentration of sulfur compounds according to storage type of garlic extract

0 One 2 3 4 A sulfide (%) 0.53 1.06 3.09 4.87 6.18 disulfide (%) 56.22 27.82 25.25 38.22 26.48 trisulfide (%) 36.31 56.97 18.04 17.42 16.91 Total amount of sulfur compounds (ppm) 511.226 201.84 133.326 52.93 66.33 B sulfide (%) 0.65 0.67 1.48 0.98 1.33 disulfide (%) 56.06 45.20 36.58 40.27 32.32 trisulfide (%) 36.23 45.01 48.61 50.82 49.45 Total amount of sulfur compounds (ppm) 615.72 202.82 510.69 350.81 231.78 C sulfide (%) 0.67 0.65 1.33 1.44 0.69 disulfide (%) 58.09 50.23 44.85 48.29 46.66 trisulfide (%) 37.17 41.91 45.64 44.47 46.51 Total amount of sulfur compounds (ppm) 670.376 270.15 657.027 620.00 461.23 D sulfide (%) 1.1 2.17 1.82 0.88 disulfide (%) 64.20 58.60 61.62 62.15 trisulfide (%) 28.11 33.52 29.01 30.25 Total amount of sulfur compounds (ppm) 524.75 675.397 683.42 706.96 E sulfide (%) 0.99 5.83 1.86 1.14 disulfide (%) 64.22 57.82 59.65 53.28 trisulfide (%) 26.39 27.02 30.19 35.25 Total amount of sulfur compounds (ppm) 506.45 485.292 503.28 501.75 F sulfide (%) 1.32 4.25 8.17 11.79 disulfide (%) 63.58 46.06 43.49 36.96 trisulfide (%) 27.12 38.26 37.42 40.04 Total amount of sulfur compounds (ppm) 451.78 443.067 539.88 565.61

A: Liquid garlic extract at room temperature

B: Liquid garlic extract, refrigerated (2 ℃)

C: Liquid garlic extract, frozen (-20 ℃)

D: Freeze-dried powdery garlic extract, frozen (-20 ℃)

E: Garlic extract in freeze-dried powder state, refrigerated (2 ℃)

F: Freeze-dried powdery garlic extract, stored at room temperature

According to these results, it was shown that garlic sulfur compounds could be stored for a long time without any change in the total amount of sulfur compounds and the proportion of sulfur compounds when stored at 2 ° C or lower under powder condition. Here, based on experimental data at 2 ° C, similar results were obtained when the storage temperature was increased to 4 ° C.

&Lt; Experimental Example 4 > Culture of bacteria against garlic extract

In order to examine the sterility of the garlic extract prepared according to one embodiment of the present invention, a culture test was conducted and the result of the culture test was shown in FIG. The extracts of garlic extracts at 1, 2, and 3 months after the extraction were prepared as the differentiation media for blood and McConkey, respectively, and then cultured for 72 hours. As shown in the figure, it was found that the growth of the bacteria did not occur regardless of the elapsed time after the extraction.

<Experimental Example 5> Effect of Garlic Extract on Cancer Cells

The garlic extract prepared according to one embodiment of the present invention was divided into various kinds of cancer cells at various concentrations to observe the effects of sulfur compounds on cancer cells.

A. Experimental Method

Garlic extract (manufactured by using 40% ethanol as 10,000 ppm) was used, and 2-fold serial dilutions were made to obtain a concentration of garlic extract of 500 μg / ml (2% ethanol) 0.016% ethanol), and the control group according to the ethanol concentration was obtained. Ellipticin was used as a positive control. 10 ml of each of the garlic extracts was placed in a 96-well plate at a constant concentration, and then various human cancer cells were placed in a 96-well plate at 190 ml of 5 to 7.5 × 10 ³ cells / well After incubation, the cells were cultured in a 5% CO ₂ incubator at 37 ° C for 72 hours, and 50% trichloroacetic acid (TCA) solution was added thereto. The cells were allowed to stand at 4 ° C for 30 minutes to 1 hour, To fix the cells. Then, a solution of 0.4% sulforhodamine B (SRB) was added thereto, and the mixture was allowed to stand at room temperature for 30 minutes to 1 hour, followed by washing with 1% acetic acid solution and water to stain fixed cells. The stained cells were dissolved in 200 mL of a 10 mM Tris solution (2-Amino-2-hydroxymethyl-propane-1,3-diol, pH 10.0) and absorbance was measured at 515 nm. Then, the survival rate (% of survival) was calculated by absorbance of each concentration of the test sample in comparison with the absorbance of the control group treated with vehicle (ethanol), and Tablecurve ver. IC ₅₀ , a concentration that inhibits 50% cell proliferation, was determined by non-linear analysis using a 1.0 program.

B. Experimental Results

1) FIG. 2 is a photograph for comparing the degree of cell proliferation after 72 hours of culturing the cancer cell line treated with the garlic extract according to the present invention and the control group treated with the beckle, respectively. As shown in the figure, On the other hand, in the well-plate containing garlic extract, cell proliferation was markedly suppressed up to a concentration of 31.39 / / ml, and the color could be confirmed. Here, the concentration of 31.39 ㎍ / ml is the fifth plate on the leftmost line in the photograph.

2) After obtaining the IC ₅₀ (50% inhibitory concentration, 50% inhibitory concentration of cell growth)% inhibition concentration of each concentration of the control of cell growth (% of inhibition) for each cancer cell lines IC ₅₀ by using the table curve And the results are shown in Table 4 and FIG. 3, respectively.

[ Table 4 ] IC ₅₀ for each cancer cell line

Cell type Garlic extract (/ / ml) Ellipticine (μM) positive control Human colon cancer cells HCT116 6.9 0.7 SW480 41.9 1.1 Human stomach cancer cells SNU638 7.0 1.0 Human lung cancer cells A549 48.0 0.6

As shown in the figure, the cancer cell growth inhibitory effect (IC ₅₀ ) of the garlic extract against four kinds of cancer cell lines was found to be in the range of 6.9 to 48.0 占 퐂 / ml. This indicates that garlic extract by the above extraction method is effective in inhibiting proliferation of stomach cancer cells and colon cancer cells.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments. Accordingly, the scope of protection of the present invention should be construed in accordance with the appended claims, and should not be construed as being limited thereto.

FIG. 1 is a photograph showing the results of the culture for 72 hours for each of the garlic extracts extracted in accordance with one embodiment of the present invention, corresponding to the elapsed time,

FIG. 2 is a photograph for comparing the degree of cell proliferation after 72 hours of culturing the cancer cells treated with the garlic extract according to the present invention and the control group treated with the vehicle,

FIG. 3 is a graph showing the survival rate (% of survival) of cancer cells after the garlic extract according to an embodiment of the present invention is applied to the cancer cells at the respective concentrations.

Claims (5)

Preparing a garlic mixture by adding ethanol to the crushed garlic, or by mixing and crushing garlic and ethanol; Aging the garlic mixture; Removing the garlic residue from the aged garlic mixture and obtaining a garlic extract; Removing the protein and polyphenol from the garlic extract; Extracting only components having a molecular weight of 5,000 or less in the garlic extract from which the protein and polyphenol have been removed; &Lt; / RTI &gt; wherein the method comprises: The method according to claim 1, And lyophilizing the garlic extract having the molecular weight of 5,000 or less and pulverizing the garlic extract. The method according to claim 1, Wherein the step of removing the protein and the polyphenol from the garlic extract comprises: Wherein the filtered garlic extract is subjected to primary filtration using a filter having a pore size of 10 to 30 占 퐉, and the filtered garlic extract is subjected to secondary filtration again using a filter having a pore size of 0.1 to 0.5 占 퐉. A garlic extract prepared by the method of claim 1 or 2, wherein the protein and the polyphenol are removed. A method for storing garlic extract, wherein the medicines using the garlic extract prepared by the method of any one of claims 1 to 3 are kept at a temperature of 4 ° C or lower in a shaded space.

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