WO2017065393A1 - Environmentally friendly alcohol-free water-soluble propolis preparation method using natural honey - Google Patents

Abstract

The present invention relates to a method for preparing environmentally friendly alcohol-free water-soluble propolis comprising: after removing foreign substances and floating materials such as wax contained in proplis, isomerizing and alkalifying honey for water-solubilization of the propolis; stirring, filtering and heating the same; and water-solubilizing the propolis by increasing the fructose content by inserting mineral water and L-arginine.

Description

Eco-friendly non-alcoholic water-soluble propolis manufacturing method using natural honey

The present invention relates to a method for producing an environmentally friendly non-alcoholic, water-soluble propolis using natural honey, and is characterized in that it produces a water-soluble propolis using the properties of fructose (Fructose) present in natural honey. Natural honey is one of the high sugar concentration foods that contain monosaccharides such as fructose and glucose and is easily absorbed by the body to become an energy source. Natural honey contains about 60 to 70% by weight of invert sugar, such as fructose and glucose, and the higher the content of glucose, the easier the crystal is made. On the contrary, the higher the content of fructose, the more the crystal is inhibited. Honey produced in Korea is generally composed of 36 to 43% by weight of fructose, 22 to 30% by weight of glucose and 3 to 4% by weight of disaccharide, sucrose. In particular, since honey has a higher content of fructose than other foods, most of honey's properties such as sweetness, hygroscopicity, and viscosity are represented by fructose. These fructose properties become stronger at lower temperatures. In addition, fructose is rapidly soluble in water, and the dissolved aqueous solution does not form a supersaturated solution, and thus does not easily form crystals, thereby increasing the dispersion coefficient and thus may be used to accommodate natural extracts. The present invention has developed a technique for receiving propolis using the above properties of honey.

Propolis, also called Russian penicillin or natural penicillin, is a material made by honeybees collecting resin from plants and mixing their secreted saliva with wax. Vegetable resins, waxes, essential oils, pollen, flavonoids and other organic substances And minerals. Bees apply the above propolis to the honeycomb gaps to protect themselves from germs and viruses and to prevent invasion by enemies such as wasps and mice. In addition, it maintains a habitat suitable for spawning and growing young bees and ripening and storing honey. Especially when spawning queen bees, it is also used for disinfection of spawning grounds.

Propolis has long been used by humans as an anti-pesticide, as evidenced by its use in Egypt in about 3000 BC. Recently, in 1965, French physician Remy Showbang, while studying honey bee-free bacteria, discovered that propolis was a natural antibiotic.

Propolis contains the most organics and minerals, and contains 104 kinds of ingredients. Among these components, minerals, vitamins, amino acids, fats, organic acids, flavonoids, etc. play an important role in cell metabolism, terpenes, etc. have an anticancer action. In particular, it contains more than 100 kinds of flavonoids, which greatly help the health of the human body.

The main benefits are anti-inflammatory, antioxidant, and immune boosting. The anti-inflammatory effect is due to half the enzymes that make prostagdins that cause inflammation in the body. In addition, flavonoids, the main ingredient, eliminates free radicals, which also have antioxidant effects. Quercetin, which has anti-cancer effects, also plays a role in blocking cancer cell genes before they are replicated.

Therefore, the demand for the propolis is expected to increase due to the availability, and it is difficult to completely remove the wax or resin, and also has problems such as being limited to use for various purposes due to being insoluble in water.

In order to solve the above problems, methods of extracting or accepting propolis have been developed. For example,

In Korean Unexamined Patent Publication No. 1997-0039866, the propolis ingot was extracted with 95% ethanol, concentrated under reduced pressure, and 5-10 times the weight of the raw material was added, and 10-20% of the total weight of the emulsifier was added to be homogeneous. A method of producing a water soluble gel propolis by concentration under reduced pressure after mixing is described.

In addition, in Korean Patent Publication No. 2002-76979, propolis ingot was extracted with 95% ethanol, water was added to make a solution of 65-70%, and mixed with sodium citrate to have a pH of 6-7, to remove wax. A method for producing a water-soluble propolis with propylene glycol after concentration under reduced pressure is described, and Japanese Laid-Open Patent Publication No. 2001-275587 describes a method for extracting and preparing a 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 preparing a propolis extract having a high water solubility by adding cyclodextrin to an extract of propolis that is difficult to be water-soluble. No. 1 shows a method of preparing a propolis extract using ethanol and using it as a honeycomb fragrance.In Korea Ginseng Research Institute, a method of preparing a propolis extract having a high solubility by adding polysorbate and lecithin (Korean Patent Publication No. 10) -1998-0022075).

In Korean Patent No. 10-486922, a method for producing an alcohol-free water-soluble propolis is mixed with propolis ingot and drinking ethanol in a ratio of 1: 4 by weight and added to the inside of the tank, while stirring with a stirring blade. Extracting and aging for about 7 to 14 days, filtration of the extracted and matured extracts first using a micro filter, and preserving the first filtration extracts in a freezer at minus 20 degrees Celsius for 48 hours. Performing a dewaxing process for separating fat, wax and resin components from the extract, and using a micro filter for the extract to remove the fat, wax and resin components separated through the primary dewaxing process from the extract. There is disclosed a manufacturing method having a step of performing a secondary filtration process, but still using alcohol Because it is extracted, there is no limit to the complete removal of alcohol at the source. Korean Patent No. 10-748846 discloses a method for producing a propolis fermented product, which is prepared by adding propolis extract powder to water, inoculating microorganisms, and fermenting the same to obtain a propolis fermented product.

Korean Patent No. 10-0601395 discloses a method for extracting propolis ingot with alcohol or alcohol, and Korean Patent Laid-Open Publication No. 2001-0017765 discloses a water-soluble pro, using sodium citrate as a PH regulator and propylene glycol as a solvent. A method for preparing polis has been disclosed. However, the above-described conventional techniques have a problem in that the manufacturing process is complicated and takes a long time by solubilizing propolis using ethanol or spirits, or accepting it through a fermentation step. .

Accordingly, the present inventors have developed a method of extracting propolis, from which wax has been removed, with ethanol such that the content of flavonoids is 2% by weight or more, and then adding L-arginine to Korean Patent Registration No. 10-1403511. The present invention has been completed by discovering that propolis can be solubilized using fructose of honey as a further improvement of the above-described prior art of the present invention.

The present invention solves the problem of the process of extracting propolis from the conventional ethanol and removes foreign substances such as foreign substances and waxes contained in the propolis, and then, in the solubilization step, the honey content is obtained through isomerization and alkalizing of honey. It is an object of the present invention to provide a method for producing an environmentally friendly, non-alcoholic water-soluble propolis that has a high solubility in propolis.

In order to achieve the above object, the present invention undergoes the following steps.

1, step to increase fructose content through isomerization and alkali treatment to take advantage of the properties of natural honey

2. Complete removal of wax and resin from propolis

3. The non-alcoholic water-soluble propolis is composed of manufacturing steps,

Stage 1. Isomerization and Alkalization of Natural Honey

1) Select natural honey that meets the food code standards.

2) Add natural honey and ethanol to the stirring tank at a weight ratio of 5-7: 5-3.

3) Stir for 3 to 5 hours at 30 ℃ or less and under 500 ~ 1000rpm conditions.

4) Add 0.5% ~ 1.0% of isomerase (ionized phosphoric acid) by weight ratio and 1.0% ~ 5.0% of L-arginine and stir at 500 ~ 1000rpm at 20 ~ 25 ℃ for 12 hours.

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

Step 2. Steps to remove wax and resin from propolis

1) Select propolis that meets the standards of health functional food standards.

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

3) Stir for 7-8 hours while heating at 40 ~ 60, 1,000 ~ 2000rpm.

4) After stirring, the mixture was rapidly cooled and left to stand for 12 to 24 hours, after which the upper layer was removed.

5) Stir for 12 to 24 hours while heating at 40 ~ 60 ℃, 1,000 ~ 2000rpm conditions again.

6) Filtration to 150 mesh or less.

Step 3. Alcohol Free Water Soluble Propolis

1) Input the 1st stage resultant and 2nd stage resultant to the stirring tank at the weight ratio of 5 ~ 20: 95 ~ 80.

2) Inject bottled water in a ratio of 1: 1 by weight ratio (in this case, the middle rain ratio means the results of the first and second stages)

3) L-Arginine is added 1.0 to 10% by weight in 2 to 3 times the weight ratio. (In this case, Jungnbi ratio means the result of 1st and 2nd stage)

4) Stir for 12 to 24 hours while heating at 40 ~ 60 ℃ under 1,000 ~ 2000rpm condition.

5) Prepare an alcohol-free water-soluble propolis while fully recovering the spent ethanol until there is no residual ethanol.

The present invention is eco-friendly by using any fructose contained in honey to propolis by using no chemical additives other than natural L-arginine, especially in the manufacturing process is not discharged waste water or pollutants at all, In addition, the propolis extracted by the method of the present invention does not retain ethanol, so the taste and aroma is soft when ingested, and palatability is improved. Since it is well soluble in water by L-arginine, dilution in water is possible for eating and eating. Bright and transparent, there is no residue left in the manufacturing process has the effect of significantly improving the yield.

1 is a culture photograph showing the activity of the control group A and samples B, C, D against Pseudomonas aeruginosa (Pseudomonas aeruginosa).

Figure 2 is the result of measuring the number of Pseudomonas aeruginosa.

Figure 3 is a culture photograph showing the activity of Control A and Samples B, C, D in E. coli.

4 is a result of measuring the number of live bacteria of E. coli.

The present invention is subjected to the following steps as a method for producing an environmentally friendly alcohol-free, water-soluble propolis.

1.Raising the content of fructose in honey through isomerization and alkalizing to take advantage of the properties of natural honey

2. Complete removal of wax and resin from propolis

4. It comprises a step of preparing an alcohol-free water-soluble propolis, if described in more detail

<Stage 1>. Isomerization and Alkalization of Natural Honey

1) Select natural honey that meets the food code standards.

2) Add natural honey and ethanol to the stirring tank at a weight ratio of 5-7: 5-3.

3) Stir for 3 to 5 hours at 500 ~ 1000rpm conditions below 30 ℃.

4) 0.5 ~ 1.0% by weight of isomerase (ionized phosphoric acid), 1.0 ~ 5.0% by weight of L-arginine and 12 hours at 500 ~ 1000rpm conditions from 20 to 25 to make the fructose content of honey more than 60% by weight After stirring, the mixture is filtered through a 100 mesh sieve.

<Step 2> Removing Wax and Resin from Propolis

1) Select propolis that meets the standards of health functional food standards.

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

3) Stir for 7-8 hours while heating at 40 ~ 60 ℃ to 1,000 ~ 2000rpm condition.

4) After stirring, the mixture was rapidly cooled and left to stand for 12 to 24 hours, after which the upper layer was removed.

5) Stir for 12 to 24 hours while heating at 40 ~ 60 ℃ to 1,000 ~ 2000rpm conditions.

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

<Step 3> Production step of alcohol-free water-soluble propolis

1) Put the 1st stage result and the 2nd stage result in the stirring tank at a weight ratio of 5 ~ 20: 95 ~ 80.

2) Pour bottled water at a ratio of 1: 1 by weight.

3) Prepare 1.0 ~ 10% by weight of L-arginine with respect to the weights of the 1st and 2nd stage products and divide into 2 ~ 3 times.

4) Stir for 12 to 24 hours while heating at 40 ~ 60 ℃ under 1,000 ~ 2000rpm condition.

5) Prepare an alcohol-free water-soluble propolis while fully recovering the spent ethanol until there is no residual ethanol.

The total flavonoid content occupying most of the phenolic compounds, which are the main constituents of the eco-friendly alcohol-free and water-soluble propolis, prepared as described above, was determined according to the Health Functional Food Code (propolis extract equivalent to about 60 to 100). Weighing (Capacity) and 90% ethanol 20 were added to dissolve and centrifuged (3,000 rpm, 10 min). Only the supernatant was taken, and the residue was washed three times with 80% ethanol 8, extracted, and the total amount of the total extract was 80% using 80% ethanol to prepare a test solution.

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

Experimental Example 1. Antimicrobial Activity

Antimicrobial activity analysis experiment was conducted by functional food research institute affiliated with Korea Health Functional Food Association, Seongnam-si, Gyeonggi-do.

As a control (A), sterile distilled water was used. Sample B is an ethanol extract propolis extract extracted with 70% ethanol and propolis in a weight ratio of 5: 1, which is the most common method for producing a propolis extract, and sample C is a commercially available British non-vitamin propolis liquid (UK Nature's). Laboratoty Ltd, Lot No.0276), and Sample D is an environmentally friendly, non-alcoholic water-soluble propolis prepared using natural honey, which is the method of the present invention, and has a total flavonoid content of 15 mg / g. Each of these samples was tested for antimicrobial activity against Pseudomonas aeruginosa (Pseudomonas aeruginosa) and Escherichia coli (E. coli).

Samples B and C of the samples were pretreated with 15 mg / g of flavonoids, the propolis indicator component, in the same manner as Sample D, by diluting the concentrations according to the KFDA. Based on the general antimicrobial test culture method, it was performed in two ways: solid culture method and liquid culture method.

In the solid culture method, each of the strain culture solution is coated on the NR agar medium, which is a microbial medium, and the paper disc, which is filled with the test sample, is incubated at 30 ° C for 30 hours, and then there is an inhibition zone around the paper disc. Observation of the size confirmed the antimicrobial activity. All tests were repeated three times.

In the liquid culture method, 1% (v / v) of the test strain culture solution and 1% (v / v) of the treated sample were simultaneously inoculated into sterilized NR liquid medium, and then incubated for 30 hours while stirring at 180 rpm at 30 ° C. After confirming the bacteria grown with the naked eye, diluted with sterile distilled water at an appropriate magnification, smeared in NR agar medium for viable cell count and incubated for 30 hours at 30 ℃ and counted the viable counts compared with the control. All experiments were repeated three times.

The medium used was NR (Nutrient Rich) medium, and the medium composition was beef extract 3.0g, peptone5.0g, Agar15.0g.

The experimental results are shown in the following [Table 1].

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

According to the above experimental results, in the case of sample D according to the present invention, growth-lowering ring formation was distinct, and the size of growth-lowering ring was 1.5 to 2 times larger than that of sample B and sample C, and similar results were obtained in measuring the number of viable cells. I could confirm it.

Sample B had a somewhat antimicrobial inhibitory effect, but it was confirmed that the antimicrobial activity is significantly lower than that of Sample D.

As shown in FIGS. 1 and 3, sterile water and sample C had weak growth growth ring formation for both Pseudomonas aeruginosa and Escherichia coli, and as shown in FIGS. 2 and 4, the number of Pseudomonas aeruginosa was significantly increased, indicating that the antibacterial activity was very weak.

The active ingredient of Sample D according to the present invention was found to have a remarkably excellent inhibitory activity against pathogenic bacteria such as Pseudomonas aeruginosa and E. coli.

Experimental Example 2. Color Comparison of Compositions Prepared by the Present Invention

Propolis liquid (tink) dietary supplement is a common intake method of diluting a certain amount of water intake. Therefore, most of domestic and foreign products do not remove resin or wax properly when diluted in water, and because of poor water solubility, the color becomes hazy, and precipitation or resin and wax floats in the upper layer to form a film. And palatability, such as clumping, is greatly reduced.

When the composition according to the present invention is diluted in water, there is no suspension phenomenon and waxes, resins, etc. do not float, thereby showing a clear, bright tan propolis intrinsic color.

To compare this, we bought four famous imported goods that are sold in department stores and compared them.

Samples are as follows.

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

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

(C) Unique water soluble propolis (product made by the technology of the present invention)

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

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

As a result of diluting each sample in 75ml bottled water based on daily intake, and confirming the results, the products (A), (B), (D), and (E), except the composition (C) of the present invention, are clouded and suspended. On the other hand, the color of the product of the present invention was clear and bright, and the color of the propolis was stabilized to a tan, a unique color of propolis, it was confirmed that the clear and bright.

Due to the usefulness of propolis, which has anti-inflammatory effects such as anti-inflammatory, anti-oxidation, and immunity enhancement, the demand is expected to increase, but it is difficult to completely remove wax or resin and is insoluble in water. Had problems such as.

The present invention is eco-friendly by using a fructose contained in honey to solubilize propolis, so that other chemical additives are not used in addition to natural L-arginine, and in particular, no waste water or pollutants are discharged during the manufacturing process, Taste and aroma are improved, and palatability is improved, and it is well dissolved in water by L-arginine, so it can be eaten by diluting with water when ingested, and the color is bright and transparent. By producing a propolis, it can be usefully used as a composition for functional foods and pharmaceuticals.

Claims (2)

Stage 1. Isomerization and alkalizing step of natural honey;

Putting the natural honey and ethanol in a stirring tank at a weight ratio of 5 to 7: 5 to 3, the first step of stirring for 3 to 5 hours at 500 ~ 1000rpm conditions at 30 ℃ or less;

0.5 to 1.0% by weight of isomerization phosphoric acid and 1.0 to 5.0% by weight of L-arginine are added to the first stirring mixture, and 500 ~ 1000rpm condition at 20-25 ° C. to make the fructose content of honey more than 60% by weight. After stirring for 12 hours with a second step of filtering through a 100 mesh (Mesh) sieve;

Propolis and ethanol were added to a stirring tank at a weight ratio of 6-8: 4-2, stirred for 7-8 hours while heating at 40-60 ° C at 1,000-2000 rpm, and then rapidly cooled, and then left for 12-24 hours. The third step of removing the filtered and stirred for 12 to 24 hours while heating to 40 ~ 60 ℃ 1,000 ~ 2000rpm conditions again and filtered through a sieve of less than 150 mesh (Mesh);

The resultant of step 2 and the resultant of step 3 were added to the stirring tank at a weight ratio of 5 to 20: 95 to 80, and bottled water and L-arginine were added again. Fourth step to do; Method for producing an alcohol-free water-soluble propolis, characterized in that made.

The method of claim 1,

In the fourth step, the content of L-arginine is 1.0 to 10% by weight based on the weight of the results of the second and third steps, the method for producing an alcohol-free water-soluble propolis.

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