KR100550165B1 - Process for preparing water soluble propolis - Google Patents

Abstract

The present invention relates to a method for producing a water-soluble propolis, and in detail, the propolis ingot is immersed in alcohol and irradiated with ionization energy and extracted to prepare a non-alcoholic propolis extract powder, and an alkali and an organic acid are added thereto. It is freeze-dried to produce a highly water-soluble propolis.

Propolis according to the present invention not only improved the extraction rate and the content of the flavonoids as an active ingredient, but also eliminates the inconvenience of use due to the inherent adhesion of the propolis extract, is easy to drink, and completely dissolved in water, so that no precipitate is formed. By having it can be used easily as a raw material of health functional food and processed food.

Description

MANUFACTURING METHOD OF WATER-SOLUBLE PROPOLIS             

1 is a view showing a ethanol extraction process according to the present invention.

2 is a view showing a process for producing a water-soluble propolis according to the present invention.

3 is a graph showing the extraction rate of the propolis extracted after irradiating the ionization energy of 3 ~ 30 kGy to the alcohol solution of the propolis ingot of the present invention.

Figure 4 is a graph showing the content of the active ingredient (flavonoid) of the propolis extracted after irradiating the ionization energy of 3 ~ 30 kGy to the alcoholic solution of the propolis ingot of the present invention.

Figure 5 is a graph showing the lipid peroxidation inhibitory effect in the liver tissue of the mouse to the water-insoluble propolis and water-soluble propolis irradiated with ionization energy of 3 ~ 7 kGy of the present invention.

The present invention relates to a method for producing a water-soluble propolis, and in detail, the propolis ingot is immersed in alcohol and irradiated with ionization energy and extracted to prepare a non-alcoholic propolis extract powder, and an alkali and an organic acid are added thereto. It is freeze-dried to produce a highly water-soluble propolis.

Propolis is a material made by bees collecting resin from plants and mixing their secretion with saliva and wax. It is composed of vegetable resins, waxes, essential oils, pollen, flavonoids and other organic substances and minerals. . The main action of propolis has various physiological activities such as antimicrobial, anti-inflammatory, analgesic, anesthetic, immune, antioxidant, antiviral, anticancer, anti-HIV and anti-AIDS. It is reported that this is due to the flavonoids that are phenolic compounds derived from the plants contained.

In Dongbogam, propolis is recorded under the name of 'Nobongbang (露 蜂房)' and explains that it is effective in relieving and asthma.In modern times, various kinds of colds, allergy, atopy, asthma, burns, skin care, anti-aging, etc. It has been used a lot as a folk medicine for immune diseases. Recently, demand is expected to increase as health and aging concerns and health functional food legislation promulgated as a public ingredient (January 31, 2004).

As described above, the increase in demand according to the human benefits of propolis is expected, but many problems have been suggested for use in various applications due to the extraction method of the propolis and inconvenience in use and insolubility after use.

Conventional propolis extraction method using alcohol (ethanol), water, glycerin, etc., even though it is not well dissolved in water but dissolved in water using a fat-based emulsifier has low absorption rate and digestive disorders have little popularity. That is, in the past, the propolis extracted with alcohol has low digestibility and low absorption rate, does not dissolve well in water, is very sticky, and once it gets on the skin, it is not erased, its taste is strong, and its fragrance is not easy to be consumed by the general public. In addition, the extraction with water has a low flavonoid content of about 0.2%, which is an active ingredient of propolis, so that the antibacterial and antioxidant functions inherent in propolis are hardly exhibited.

Therefore, a lot of research is in progress to improve this problem.

In Korean Patent Laid-Open Publication No. 1999-75441, propolis ingot was extracted with 95% ethanol, concentrated under reduced pressure, added 5-10 times of the weight of raw material, and added 10-20% of emulsifier to make it homogeneous. A method of producing a water soluble gel propolis by concentration under reduced pressure after mixing is described. Korean Patent Publication No. 2002-76979 discloses extracting propolis with 95% ethanol, adding water to make a solution of 65 ~ 70%, and mixing it with sodium citrate so that the pH is 6 ~ 7, removing wax and concentrating under reduced pressure. A method for preparing a water soluble propolis with propylene glycol is then described. Japanese Patent Laid-Open No. 2001-275587 describes a method for extracting and preparing a propolis stock solution using far infrared rays at low temperature.

However, the Republic of Korea Patent Publication No. 1999-75441 recognizes the simplicity of the manufacturing process, but the use of 10 ~ 20% of the fat emulsifiers (polysorbate, monoglycerin acetate, lecithin) is reduced in absorption rate and digestive problems In addition, Korean Patent Publication No. 2002-76979 used propylene glycol as an aqueous solvent of propolis in Example A-1, which causes changes in physical properties due to layer separation due to instability of fatty components during long-term storage. The emulsifier (glycerine) added in consideration of stabilization of the fat component in Example A-2 is a method that has not been considered popular because it inhibits the active effect of the propolis extract and the absorption rate of the human body. In addition, the use of activators and non-sweetening agents added to compensate for these shortcomings seems to be somewhat uneconomical. In addition, Japanese Patent Application Laid-Open No. 2001-275587 is an economical extraction method capable of extracting propolis stock in a short time, but the water-insoluble propolis stock is expected to lower the absorption rate when taken.

In order to solve the above problems, the present inventors prepared the water-soluble propolis without any emulsifier after extracting it by irradiating ionized energy on the propolis ingot, and the extraction rate and the content of flavonoids contained in the propolis extract were increased. Confirmed that the lipid peroxidation inhibitory effect was completed the present invention.

The present invention is to provide a method for producing a water-soluble propolis without using any emulsifier after extracting by irradiation with ionization energy to the propolis ingot.

The present invention provides a method for producing a water-soluble propolis.

Hereinafter, the present invention will be described in detail.

The manufacturing method of the present invention

1) grinding the propolis ingot and dissolving it in alcohol, and then extracting by irradiating ionization energy (first step); And

2) adding water to the propolis extract powder prepared in the first step and then adding the alkali and the organic acid to solubilize it (second step).

In the first step, the pulverized propolis ingot was immersed in alcohol and immersed, and then stirred for 1 to 2 hours. The solution was irradiated with 3 to 30 kGy at 10 to 20 Gy / min at room temperature to 35 to 40 ° C. After extracting for 4-6 hours in filtration, concentration and freeze-dried to prepare an alcohol-free propolis extract powder. The ionization energy includes radiation such as α, β, and γ rays, and γ rays are most preferred in the present invention.

In the second step, the volume ratio of water, propolis powder and alkali is set at 10 to 15: 1: 0.1 to 0.8 and stirred at room temperature for 1 to 2 hours. The alkali serves to convert the hydrophobic group of the propolis into a hydrophilic group to form a water-soluble propolis, and the alkali includes sodium hydroxide, ethanolamine, potassium hydroxide, sodium carbonate, and the like, and sodium carbonate is most preferred in the present invention.

1 to 6 ml of organic acid is added to the mixture, and the pH of the solution is adjusted to 6.0 to 7.0 by stirring at room temperature for 2 to 3 hours. Organic acids are used to adjust the pH of the propolis solution to neutral. Organic acids include formic acid, lactic acid, citric acid, acetic acid and the like, and citric acid is preferred in the present invention.

The solution is concentrated and lyophilized to obtain an alcohol-free, water soluble propolis extract powder.

In the present invention, the degree of solubilization varies depending on the ratio of alkali and citric acid. In other words, if the amount of alkali used for solubilization is added in a smaller proportion than the propolis extract powder, the degree of dissolution is decreased. If the amount is added in excess, the degree of dissolution decreases. This is because the water soluble propolis is reduced to the water insoluble propolis.

The extraction rate of the propolis extract according to the present invention is 42% for the irradiated ionization energy (0 kGy), 60 to 62% irradiated for 3 to 30 kGy, 18 to 20% extraction of the propolis irradiated with ionization energy Appear higher.

In addition, the flavonoid content of the propolis extract according to the present invention appears to be 4.6 mg / g higher than irradiated with 3 ~ 7 kGy of ionization energy than the irradiated energy, but the ionization of 8 ~ 15 kGy and 20 ~ 30 kGy The flavonoid content of the propolis extracts, which were energized, was 4.4 mg / g and 7.3 mg / g, respectively.

In addition, the degree of peroxidation of lipids in biological tissues by CCl ₄ administration of the propolis extract according to the present invention was 2 mg / of non-irradiated non-aqueous propolis samples, irradiated water-soluble and non-aqueous propolis samples (3-7 kGy). In the mouse-treated group, the control group was significantly inhibited compared to the CCl ₄ control group. In addition, when the concentration of irradiated water-soluble propolis was doubled (4 mg / mouse), the inhibitory effect was 12% higher than that of the irradiated water-soluble propolis sample administered 2 mg / mouse.

Therefore, the extraction rate of the propolis extract according to the present invention is proportional to the irradiation amount of ionization energy, the content of flavonoids as an active ingredient is the most optimal content when irradiating the ionization energy of 3 ~ 7 kGy, lipid peroxidation in liver tissue of the mouse By showing an inhibitory effect, it can be usefully used as an antioxidant.

Hereinafter, the present invention will be described in detail by examples.

However, the following examples are merely to illustrate the present invention, the present invention is not limited by the following examples.

Example Preparation of Water-Soluble Propolis

1. Extracting Propolis

Frozen propolis ingot (Chinese) in the resin state was finely pulverized with a mill and used as a sample for ionization energy irradiation and water-soluble propolis preparation.

Propolis coagulum and 80% alcohol ground in a reaction mixture was added at a volume ratio of 1: 10, and stirred at room temperature for 1 hour. A total of 10 kGy (10 Gy / min) of ionization energy was collected using a ⁶⁰ Co gamma irradiation device. It was investigated in a reaction mixer at room temperature. After irradiation, the extract was additionally extracted at 40 ° C. for 4 hours, cooled to room temperature, filtered, concentrated, and lyophilized to prepare an alcohol-free propolis extract powder.

2. Solubilization of Propolis Powder

In order to solubilize the propolis extract powder prepared in the above 1, the volume ratio of water, propolis extract powder and sodium carbonate was set to 12: 1: 1: 0.3 and stirred vigorously at room temperature for 2 hours. 6 ml of 3% citric acid was added thereto, followed by further stirring at room temperature for 3 hours. The solution was concentrated and lyophilized to prepare an alcohol-free, water soluble propolis extract powder.

In order to determine the degree of dissolution in the water-soluble propolis extract powder prepared above, the degree of dissolution was measured, and the dissolution degree was 99% or more.

Therefore, it can be seen that the ratio of sodium carbonate used for solubilizing the propolis extract of the present invention and citric acid used as a pH regulator has the greatest effect.

The ethanol extraction process diagram according to the present invention is shown in Figure 1, the manufacturing process of the water-soluble propolis according to the present invention is shown in Figure 2.

Experimental Example 1 Extraction Rate of Propolis

In order to determine the effect of increasing the extraction rate of propolis according to the present invention on the ionization energy, the following experiment was performed.

1 g of propolis ingot was pulverized, 15 ml of 80% ethanol was added thereto, and the mixture was allowed to stand for 24 hours after checking the ionization energy. Stirring for 10 minutes and centrifugation (3000 rpm, 10 minutes) to drain the clear solution and the residue was diluted with 10 ml of 80% ethanol was repeated three times. The extracts were collected so that the total amount was 50 ml. 10 ml of this solution was dried at 105 ° C. for 4 hours to obtain a propolis extract, and the sample was calculated as in Equation 1 below to obtain an extraction rate in 80% ethanol.

The results are shown in Table 1 and FIG.

  Ionization Energy Dose (kGy)   Propolis Extraction Rate (%) 0 42 3 to 7 60 8 to 15 60 20 to 30 62

As shown in Table 1 and Figure 3, the extraction rate of the propolis extract according to the present invention is not irradiated with ionization energy (0 kGy) 42%, 3-7 kGy and 8-15 kGy irradiated 60%, 62% of the 20-30 kGy irradiation showed 18-20% higher extraction rate of propolis irradiated with ionization energy.

This indicates that the condensates produced by the condensation of terpene and flavone compounds contained in the resin of propolis are hydrolyzed from the resin by interaction with ionization energy, and the extraction rate is higher as the amount of application of ionization energy increases. Able to know.

Therefore, it can be seen that the extraction rate of the propolis extract according to the present invention is proportional to the irradiation amount of ionization energy.

Experimental Example 2 Flavonoid Content of Propolis Extract

In order to determine the flavonoid content of the propolis extract according to the present invention, the following experiment was performed.

Below, quercetin (Quercetin), ethanol (Express), 10% aluminum nitrate solution, 0.1 M potassium acetate solution was used as a reagent.

Quercetin 50 mg was accurately measured and dissolved in 80% ethanol to a volume of 5 ml. Ethanol was added to the solution, diluted to 100, 20, and 10 times, and 0.1, 0.05, and 0.1 mg / mL standard solutions were prepared, respectively, and the absorbance was calculated to prepare a calibration curve. A test solution was prepared by adding and dissolving 1 ml of 80% ethanol to 10 mg of the propolis extract powder obtained in the above example, and after adding 0.1 ml of 10% aluminum nitrate solution and 0.1 ml of 0.1 M potassium acetate to 0.05 ml of the test solution. The total volume was made 5 ml with 80% ethanol, and the mixture was sufficiently stirred and allowed to stand at room temperature for 40 minutes. Absorbance was measured at 410 nm using water as a control. The flavonoid content (mg / g) was calculated by substituting the difference obtained by measuring the absorbance obtained by subtracting the absorbance obtained by adding 0.1 ml of water instead of aluminum nitrate from the measured absorbance.

The results are shown in Table 2 and FIG.

  Ionization Energy Dose (kGy)   Flavonoid Content (mg / g) 0 100.9 3 to 7 105.9 8 to 15 96.5 20 to 30 93.6

As shown in Table 2 and Figure 4, the flavonoid content of the propolis extract according to the present invention is 100.9 mg / g, 35.5 ~ 7 kGy in irradiated (0 kGy), 105.5 mg / g, 8 96.5 mg / g irradiated with 15 kGy and 93.6 mg / g irradiated with 20-30 kGy. In particular, the flavonoid content of the propolis extract irradiated with ionization energy of 3-7 kGy was 4.6 mg / g higher than that without irradiating ionization energy. However, flavonoid contents of propolis extracts with ionization energies of 8-15 kGy and 20-30 kGy were lower than 4.4 mg / g and 7.3 mg / g, respectively.

When the excessive ionization energy of 8-15 kGy or 20-30 kGy is irradiated, flavonoids, which are active ingredients of propolis, are induced by the radical reaction by interaction with ionization energy, which induces the quinone compound to reduce the flavonoid content. to be.

Therefore, the flavonoid content of the propolis extract according to the present invention can be seen that the most optimal content state when irradiating the ionization energy of 3 ~ 7 kGy.

Experimental Example 3 Inhibitory Effect of Propolis on Lipid Peroxidation

In order to find out how the water-soluble and water-insoluble propolis powder samples according to the present invention inhibit the degree of lipid peroxidation in biological tissues caused by chemical factors, the mice were administered to the mice, treated with chemicals, and the livers were extracted to obtain intracellular tissues. The degree of lipid peroxidation was measured.

Lipids present in biofilms produce malondialdehyde (MDA) as secondary byproducts oxidized by chemicals, X-rays, and radiation, which react with thiobarbituric acid (TBA). Since the color was developed, it was measured by an ultraviolet spectrophotometer.

Four mice were used in each experimental group to measure the degree of lipid peroxidation among the mice, and CCl ₄ was orally administered with 2 mg per mouse. Oral administration of water-insoluble and water-soluble propolis was administered orally 27 hours and 3 hours prior to CCl ₄ administration. One day after the CCl ₄ administration, a certain amount of liver tissue was homogenized with a homogenizer with 1.15% KCl, separated by protein by centrifugation, quantified to 3 mg, and then 8.1% sodium dodecyl sulfate ( 0.2 ml of sodium dodecyl sulfate (SDS), 1.5 ml of 20% acetic acid (pH 3.0), and 1.5 ml of 0.8% TBA solution were mixed well, and distilled water was added so that the final volume was 4 ml. After 30 minutes of hot bath at 95 ℃, cooled to room temperature and centrifuged for 10 minutes at 3500 rpm and the supernatant was taken to measure the absorbance at 532nm.

The results are shown in Table 3 and FIG.

sample liver nM MDA / mg Protein Normal control 1.1 Carbon tetrachloride 14.2 Non-irradiated (0kGy) water-insoluble sample (2 mg) administration group 6.4 Irradiation (3 ~ 7kGy) non-aqueous sample (2mg) administration group 6.2 After irradiation (3 ~ 7kGy) soluble sample (2mg) administration group 8.5 After irradiation (3 ~ 7kGy) soluble sample (4mg) administration group 6.9

As shown in Table 3 and FIG. 5, the degree of peroxidation of lipids in biological tissues by CCl ₄ administration was measured in non-irradiated non-aqueous propolis samples (0 kGy), irradiated water-soluble propolis samples (3-7 kGy) and irradiation ratios. It was found that 2 mg / mouse of the water-soluble propolis sample (3-7 kGy) was significantly inhibited 44-61% compared to the CCl ₄ control group. The inhibitory effect was as low as 5 ~ 17%. In addition, when the concentration of irradiated water-soluble propolis was doubled (4 mg / mouse), the inhibitory effect was 12% higher than that of the irradiated water-soluble propolis sample administered 2 mg / mouse.

Therefore, the water-insoluble and water-soluble propolis irradiated with the ionization energy of the present invention can be used as an antioxidant that is useful for protecting the living body since it has an effect of inhibiting oxidative damage of lipids. The difference of the inhibitory effect between the two samples is 5 ~ 17%, so there is no big difference because the range of standard error is large.

Propolis according to the present invention not only improved the extraction rate and the content of the flavonoids as an active ingredient, but also eliminates the inconvenience of use due to the inherent adhesion of the propolis extract, is easy to drink, and completely dissolved in water, so that no precipitate is formed. By having it can be used easily as a raw material of health functional food and processed food.

Claims (10)

1) grinding the propolis ingot and dissolving it in alcohol, and then extracting by irradiating ionization energy (first step); And 2) The method of producing a water-soluble propolis comprising the step (second step) of adding water to the propolis extract powder prepared in the first step and then adding an alkali and an organic acid. The method of claim 1, wherein the alcohol in the first step is 80% alcohol (ethanol). The method of claim 1, wherein the ionization energy in the first step is radiation comprising alpha (α), beta (β), gamma (γ) rays. The method of claim 3, wherein the ionization energy is gamma ray and irradiated at 3 to 30 kGy. The method of claim 1, wherein the volume ratio of water, propolis extract powder, and alkali in the second step is 10 to 15: 1: 0.1 to 0.8. The method of claim 5, wherein the alkali is selected from the group consisting of sodium hydroxide, ethanolamine, potassium hydroxide and sodium carbonate. The method of claim 6, wherein the alkali is sodium carbonate. The method of claim 1, wherein the organic acid in the second step is selected from the group consisting of formic acid, lactic acid, citric acid and acetic acid. The method of claim 8, wherein the organic acid is citric acid. The method of claim 1, wherein the pH in the second step is 6.0 ~ 7.0 method for producing a water-soluble propolis.

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