KR101661495B1 - The method of Eco-friendly alcohol-free, water-soluble propolis using natural honey - Google Patents

Abstract

The present invention relates to a method for producing eco-friendly, non-alcoholic, and water-soluble propolis using natural honey. The method of the present invention comprises the following steps: increasing the content of fructose in honey by undergoing isomerization and alkali treatment; completely eliminating wax and a resin from propolis; and producing the non-alcoholic and water-soluble propolis.

Description

[0001] The present invention relates to a process for preparing an eco-friendly alcohol-free, water-soluble propolis using natural honey,

The present invention relates to a process for producing environmentally-friendly non-alcoholic and water-soluble propolis using natural honey, and is characterized in that a water-soluble propolis is produced using the characteristics of fructose present in natural honey. Natural honey is one of the most highly concentrated foods based on monosaccharides such as fructose and glucose. It is easily absorbed in the body and becomes an energy source. Natural honey contains about 60 ~ 70% by weight of monosaccharides such as fructose and glucose. The higher the content of glucose, the easier the crystals are. In contrast, the higher the content of fructose, the more the crystals are inhibited. The honey produced in Korea generally has 36 to 43% by weight of fructose, 22 to 30% by weight of glucose, and 3 to 4% by weight of disaccharide. In particular, since honey has a higher content of fructose than other foods, it can be said that most of the characteristics of honey such as having a strong sweetness and hygroscopicity and having a viscosity are exhibited by fructose. These fructose properties become stronger at lower temperatures. The fructose is soluble in water and the dissolved aqueous solution does not form a supersaturated solution and does not easily form crystals, so that the dispersion factor can be increased and used for watering the natural extract. The present invention has developed a technique for hydrolyzing propolis using the above-mentioned characteristics of honey.

Propolis, also known as Russian penicillin or natural penicillin, is a substance made by mixing beeswax with saliva secreted from a plant by collecting a resin from a plant. The propolis is made of vegetable resin, wax, essential oil, pollen, flavonoid, And minerals. The honey bees protect themselves from germs and viruses by spraying the propolis above the cracks of the honeycomb and prevent enemies such as wasps and rats. It also maintains habitat suitable for spawning and growth of baby bees, aging and storage of honey. Especially when the queen bee is scattered, it is also used to disinfect the spawning ground.

These propolis have long been used by mankind as an anti-pesticide for so long that there is evidence that it was used in Egypt in about 3000 BC. Recently, in 1965, a French physician, Remy Shaw, discovered that propolis was a natural antibiotic while studying the absence of bacteria in bee's body.

Organic and mineral constituents of propolis are the most abundant, with about 104 ingredients. Minerals, vitamins, amino acids, fats, organic acids and flavonoids play important roles in cell metabolism and terpenes have anticancer activities. Especially, it contains more than 100 kinds of flavonoids, which is very helpful for improving the health of the human body.

Major efficacy include anti-inflammation, antioxidant, and immunity enhancement. The anti-inflammatory effect is because it reduces the enzyme that produces prostaglandin, which causes inflammation in human body, by half. The main ingredient, flavonoid, also has antioxidant effect because it eliminates active oxygen. And quercetin, which has an anticancer effect, so as to block cancer genes before their genes are replicated.

Therefore, the demand for propolis is expected to increase due to the usefulness of propolis, but it is difficult to completely remove wax or resin and is also insoluble in water.

In order to solve the above-mentioned problems, methods for extracting propolis or methods for encapsulating propolis have been developed. For example,

In Korean Patent Laid-Open Publication No. 1997-0039866, a propolis raw material is extracted with 95% ethanol and concentrated under reduced pressure. Water is added 5 to 10 times as much as the raw material weight, and 10 to 20% Followed by concentration under reduced pressure to prepare propolis in a water-soluble gel state.

Also, in Korean Patent Laid-Open Publication No. 2002-76979, a propolis raw material is extracted with 95% ethanol, water is added to make a 65-70% solution, and the pH is adjusted to 6-7 with sodium citrate to remove the wax There is disclosed a process for producing a water-soluble propolis with propylene glycol after concentration under reduced pressure, and JP-A-2001-275587 discloses a process for extracting and preparing propolis stock solution using far-infrared rays at low temperature.

In addition, Korean Patent Laid-Open Publication No. 10-1992-0014532 discloses a method for producing propolis extract in which water-solubility is enhanced by adding cyclodextrin to an extract of propolis which is difficult to be made water-soluble, and disclosed in Korean Patent Publication No. 90-7005 A process for preparing propolis extract by using ethanol in water and a method for using it as a flavor enhancer has been proposed. The Korean Ginseng Research Institute has proposed a process for producing propolis extract having enhanced solubility by adding polysorbate and lecithin -1998-0022075).

Korean Patent No. 10-486922 discloses a method for producing alcohol-free water-soluble propolis, wherein a propolis raw material and ethanol capable of drinking are mixed at a ratio of 1: 4 on a weight basis, placed in a tank, stirred with a stirring blade Extracting and aging the extracts in an amount of 7 to 14 days, filtering the extracted and extracted extracts by using a microfilter, and extracting the extracted primary extracts in a freezer at a temperature of minus 20 degrees Celsius for 48 hours A step of performing a dewaxing process for separating fat, wax and a resin component from the extracted liquid; and a step for removing the fat, wax and resin components separated through the primary dewaxing process using a microfilter A step of performing a secondary filtration step, and the like are disclosed. However, it is still possible to use an alcohol It is inevitable to completely remove alcohol from the source. Korean Patent No. 10-748846 discloses a process for producing a propolis fermented product by adding a propolis extract powder to water and fermenting the microorganism by inoculation to obtain a propolis fermented product.

Korean Patent Publication No. 10-0601395 discloses a method for extracting a propolis raw material with an alcohol or a sponge. Korean Patent Laid-Open Publication No. 2001-0017765 discloses a method of extracting propolis using an aqueous solution of propylene glycol as a pH regulator, sodium citrate as a pH regulator, However, the above conventional techniques have been problematic in that the production process is complicated and takes a long time, such as by using propolis or ethanol for the purpose of water-solubilizing propolis or fermenting it .

Accordingly, the present inventors have developed a method of extracting propolis from wax and the like with ethanol so that the content of flavonoid is not less than 2% by weight, and then adding L-arginine into the solution to make it water-soluble, as disclosed in Korean Patent No. 10-1403511, The inventors of the present invention have found that propolis can be made water-soluble by using fructose of honey, which is a further improvement of the above-described prior art.

1. Korean Published Patent Application No. 1997-39866 1. Korean Patent Publication No. 2002-76979 1. Korean Patent Publication No. 1992-14532 1. Korean Patent Publication No. 1990-7005 1. Domestic Patent Publication No. 1998-22075 1. Korean Patent Publication No. 2001-17765 1. Domestic registered patent No. 486922 1. Domestic registered patent No. 748846 1. Domestic Registration No. 0601395 1. Domestic Registration No. 1403511 1. Japanese Laid-Open Patent Publication No. 2001-275587

The present invention solves the problem of the conventional process of extracting propolis from ethanol and removes suspended substances such as wax and foreign substances contained in propolis, and then treats honey with isomerization treatment and alkalization treatment in a water-soluble step, Soluble propolis, and a method for producing an environmentally-friendly, non-alcoholic water-soluble propolis for propolis.

To achieve the above object, the present invention is directed to the following steps.

1, a step of increasing the sugar content by isomerization and alkalinization to utilize the characteristics of natural honey

2. Complete removal of wax and resin from propolis

3. Non-alcoholic water-soluble propolis consists of a manufacturing step,

Stage 1. Isomerization and alkalization of natural honey

1) Select natural honey that meets food standards.

2) Add natural honey and ethanol at a mixing ratio of 5 ~ 7: 5 ~ 3.

3) Stir for 3 to 5 hours at a temperature of 30 ° C or less, 500 to 1000 rpm.

4) The isomerization enzyme (ionized phosphoric acid) is added in an amount of 0.5% to 1.0% by weight and L-arginine in a weight ratio of 1.0% to 5.0%, and stirred at 20 to 25 ° C under 500 to 1000 rpm for 12 hours.

5) After confirming that the fructose content is 60% by weight or more, filter it with a sieve of 100 mesh or less.

Step 2. Step of removing wax and resin from propolis

1) Select propolis suitable for health functional foods standard.

2) Propolis and ethanol are added to the stirring tank at a weight ratio of 6-8: 4 ~ 2.

3) While heating at 40 ~ 60, 1,000 ~ 2000rpm, stir for 7 ~ 8 hours.

4) After stirring, cool rapidly and let stand for 12-24 hours, then remove the upper layer.

5) Stir again for 12 to 24 hours while heating at 40 to 60 ° C and 1,000 to 2000 rpm.

6) Filter under 150 mesh (Mesh).

Step 3. Nonalcoholic water-soluble propolis preparation step

1) The first stage product and the second stage product are put into a stirring tank at a weight ratio of 5: 20: 95 ~ 80.

2) Bottled water is added at a ratio of 1: 1 by weight ratio (here, the weight ratio refers to the result of step 1 and step 2)

3) L-arginine is added in two to three divided doses in an amount of 1.0 to 10% by weight based on the weight ratio (here, the weight ratio means the result of step 1 and step 2)

4) While heating at 40 to 60 ° C under 1,000 to 2000 rpm, stir for 12 to 24 hours.

5) Using a concentrator, make a non-alcoholic water-soluble propolis with complete recovery until no ethanol remains.

The present invention is advantageous in that it does not use any chemical additives other than natural L-arginine by water-soluble propolis by using fructose contained in honey, and thus is eco-friendly. Particularly, In addition, the propolis extracted by the method of the present invention does not retain ethanol, so the taste and smell are soft and improved in palatability when ingested. Since it is dissolved in water by L-arginine, it can be diluted with water to be ingested, Bright and transparent, and there is no residue left in the manufacturing process, thereby remarkably improving the yield.

Fig. 1 is a photograph showing the activity of control group A and samples B, C and D in P. aeruginosa (Pseudomonas aeruginosa).
Fig. 2 shows the result of measuring the number of P. aeruginosa.
Fig. 3 is a photograph showing the activity of the control group A and the samples B, C and D in E. coli.
4 shows the results of measurement of the number of viable cells of Escherichia coli.

The present invention relates to a process for producing environmentally-friendly non-alcoholic and water-soluble propolis.

1. Increasing the sugar content of honey through isomerization and alkalization to take advantage of the properties of natural honey

2. Complete removal of wax and resin from propolis

4. Constituting a non-alcoholic water-soluble propolis, which will be described in more detail

<Stage 1>. Isomerization and alkalization of natural honey

1) Select natural honey that meets food standards.

2) Put honey and ethanol in the mixing tank at a ratio of 5 ~ 7: 5 ~ 3.

3) Stir at a temperature of 30 ° C or less at 500 to 1000 rpm for 3 to 5 hours.

4) 0.5 to 1.0% by weight of isomerizing enzyme (ionized phosphoric acid) and 1.0 to 5.0% by weight of L-arginine were added and the mixture was stirred at 20 to 25 to 500 to 1000 rpm for 12 hours After stirring, the mixture is filtered through a mesh of 100 mesh.

&Lt; 2 &gt; Step of removing wax and resin from propolis

1) Select propolis suitable for health functional foods standard.

2) Propolis and ethanol are added to the stirring tank at a weight ratio of 6-8: 4 ~ 2.

3) While heating at 40 to 60 ° C at 1,000 to 2,000 rpm, stir for 7 to 8 hours.

4) After stirring, cool rapidly and let stand for 12-24 hours, then remove the upper layer.

5) The mixture is stirred at 40 to 60 ° C for 12 to 24 hours while heating at 1,000 to 2,000 rpm.

6) Filter with a sieve of 150 mesh or less.

<Step 3> Preparation of non-alcoholic water-soluble propolis

1) The first stage product and the second stage product are put into a stirring tank at a weight ratio of 5: 20: 95 ~ 80.

2) Bottled water is added at a ratio of 1: 1 by weight.

3) Prepare 1.0 ~ 10% by weight of L-arginine with respect to the weight of the first stage product and the second stage product, and then add them in two or three divided doses.

4) While heating at 40 to 60 ° C under 1,000 to 2000 rpm, stir for 12 to 24 hours.

5) Using a concentrator, make a non-alcoholic water-soluble propolis with complete recovery until no ethanol remains.

The content of total flavonoids, which constitute most of the phenolic compounds, which are major constituents of the environmentally-friendly non-alcoholic and water-soluble propolis prepared as described above, was determined by extracting the extract (amount corresponding to about 60-100 with propolis extract) Weighing (Capacity), weighing), 90% ethanol 20 was added thereto, dissolved and centrifuged (3,000 rpm, 10 min). The supernatant was removed, and the residue was further washed three times with 80% ethanol 8 and extracted. The total extractives were combined and the total volume was adjusted to 50 with 80% ethanol to prepare a test solution.

Test solution 0.5 was taken in a test tube, and ethanol 1.5, 10% aluminum nitrate solution 0.1, 1 M potassium acetate solution 0.1, and water 2.8 were added and sufficiently stirred. After standing at room temperature for 40 minutes, the absorbance of the liquid layer was measured at 415 nm using 10 cells. As a blank control for the sample, total flavonoid was calculated based on a calibration curve made with quercetin by adding 0.1 of water in place of the aluminum nitrate solution during the above operation as the difference in absorbance.

Experimental Example  1. Antimicrobial activity experiment

Antimicrobial activity analysis experiment was conducted by the Functional Food Research Institute of Korea Health Functional Food Association, located in Seongnam, Gyeonggi-do.

Control (A) was sterile distilled water. Sample B is an ethanol-extracted propolis extract obtained by extracting 70% ethanol and propolis at a weight ratio of 5: 1, which is the most common method for producing propolis extract. Sample C is a commercially available non-vital propolis liquid (UK Nature's Laboratoty Ltd, Lot No.0276), and Sample D is an eco-friendly, non-alcoholic water-soluble propolis prepared using natural honey as the method of the present invention, and has a total flavonoid content of 15 mg / g. Antimicrobial activity against P. aeruginosa, E. coli and P. aeruginosa was investigated using these samples.

Sample B and sample C were pretreated with 15 mg / g of flavonoid as the propolis index component in the same manner as in Test D by concentration dilution according to the Health Functional Food Test Method defined by the Ministry of Food and Drug Administration. Control group A was sterilized distilled water And two methods were used for solid culture and liquid culture based on the general antibacterial test culture method.

In the solid culture method, the culture medium of the strain was streaked on the NR agar medium, and the paper disc with the test sample was placed on the upper layer. After incubation at 30 ° C for 30 hours, the presence or absence of the inhibition zone around the paper disc The size was observed to confirm the antimicrobial activity. All tests were repeated 3 times.

In the liquid culture method, the inoculated NR liquid medium was inoculated simultaneously with 1% (v / v) of the test strain culture medium and 1% (v / v) of the treated sample and incubated for 30 hours at 30 ° C with stirring at 180 rpm. After incubation at 30 ° C for 30 hours, the number of viable cells was counted and compared with that of the control group. The number of viable cells was counted after sterilization with sterilized distilled water. All experiments were repeated 3 times.

The medium used was NR (Nutrient Rich) medium. The medium composition was 3.0 g of beef extract, 5.0 g of peptone and 15.0 g of Agar.

The experimental results are shown in Table 1 below.

sample Pseudomonas aeruginosa (water monosus,
Pseudomonas aeruginosa) E. coli Control A - - Sample B ++ ++ Sample C + + Sample D ++++ +++

According to the results of the above experiment, the growth inhibition ring formation of the sample D of the present invention was remarkable and the size of the growth inhibition ring was 1.5 to 2 times larger than that of the sample B and the sample C, I could confirm.

Sample B showed some antimicrobial inhibitory effect, but it was confirmed that the antibacterial activity was significantly lower than that of Sample D.

As shown in FIGS. 1 and 3, the sterilized water and the sample C showed weak growth inhibition against both Pseudomonas aeruginosa and Escherichia coli. As shown in FIGS. 2 and 4, the number of Pseudomonas aeruginosa was significantly increased and the antimicrobial activity was very weak.

It has been confirmed that the active ingredient of the sample D according to the present invention has a remarkably excellent inhibitory activity against pathogenic bacteria such as Pseudomonas aeruginosa and Escherichia coli.

Experimental Example 2   . Comparison of colors of compositions made by the present invention

Propolis liquid type (tincture type) It is a general method of ingesting a certain amount of health functional food by diluting it with water. In most domestic and foreign products, when the resin is diluted with water, the resin or wax is not properly removed. Since the resin is not sufficiently water-soluble, the color becomes cloudy, and the resin is precipitated or the resin and wax are floated on the upper layer to form a film Such as forming a lump or significantly reduce the palatability.

When diluted with water, the composition according to the present invention does not have a suspension phenomenon, and wax, resin and the like do not float, so that the composition shows a clear bright yellowish brown propolis specific color.

To compare this, four famous imports sold at the department stores were purchased and compared.

The samples are as follows.

(A) Bee Vital Propolis Liquid (UK, Nature's Laboratory ltd)

(B) Propolis Liquid (Australia, JNS Pharma Pty Ltd)

(C) Unique water-soluble propolis (product manufactured by the technique of the present invention)

(D) Comvita Proppolis Tincture (Comvita New Zealand Ltd, New Zealand)

(E) Tong Life Propolis Liquid (Canada, Triton Pharmaceutical Ltd

As shown in <Figure 1>, it was observed after 10 days, and the result is shown in <Figure 2>.

<Figure 1> Color Comparison (Day 1)

<Figure 2> Color Comparison (Day 10)

(A) Bee Vital Propolis Liquid (UK, Nature's Laboratory ltd)

(B) Propolis Liquid (Australia, JNS Pharma P Ltd)

(C) Unique water-soluble propolis (product of the present invention)

(D) Comvita Proppolis Tincture (Comvita New Zealand Ltd, New Zealand)

(E) Tong Life Propolis Liquid (Triton Pharmaceutical Ltd, Canada)

<Figure 1> shows that the products A, B, D and E except for the composition C of the present invention are suspended and cloudy, whereas C is clear and bright in color, and is stable in tan color which is inherent in propolis It was bright and transparent. According to FIG. 2, even after 10 days, the product C of the present invention did not cause floatation or sedimentation, but the products A, B, D and E were produced by the production of resin and wax, Compared with the conventional method.

Claims (3)

Stage 1. Isomerization and alkalization of natural honey;
Mixing natural honey and ethanol in a mixing tank at a weight ratio of 5 to 7: 5 to 3, and stirring the mixture at 30 ° C or less at 500 to 1000 rpm for 3 to 5 hours;
0.5 to 1.0% by weight of ionized phosphoric acid, isomerizing enzyme, and 1.0 to 5.0% by weight of L-arginine are added to the above primary agitated product and the mixture is heated at 20 to 25 DEG C at 500 to 1000 rpm to a fructose content of at least 60% Followed by filtration through a sieve of 100 mesh (Mesh);
Propolis and ethanol were added to a stirring tank at a weight ratio of 6 to 8: 4 to 2, and the mixture was stirred at 7 to 8 hours while being heated at 40 to 60 ° C at 1,000 to 2,000 rpm, rapidly cooled, allowed to stand for 12 to 24 hours, , Stirring the mixture for 12 to 24 hours while heating the mixture at a temperature of from 40 to 60 ° C at a temperature of from 1,000 to 2,000 rpm, and then filtering the mixture through a sieve having a mesh size of 150 mesh or less;
The second stage product and the third stage product were charged into a stirring tank at a weight ratio of 5 to 20:95 to 80, and then water and L-arginine were added thereto. The mixture was heated at 40 to 60 ° C at 1,000 to 2,000 rpm for 12 to 24 hours The fourth system; Soluble propolis. &Lt; / RTI &gt;
delete The method according to claim 1,
Wherein the content of L-arginine in the fourth step is 1.0 to 10% by weight based on the weight of the result of the second step and the third step of the non-alcoholic water-soluble propolis.

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