KR101070382B1 - Soap containing the extracts of Zizyphus jujube seed and manufacturing method of it - Google Patents

Abstract

The present invention relates to a soap using a jujube seed extract and a method for manufacturing the same, and to provide a soap and a method for producing the jujube seed extract concentrate, which is concentrated under reduced pressure of the ethanol extract of jujube seed, in a total weight of 0.5-10% by weight, While reusing seeds, the extract is obtained using ethanol having excellent cell membrane permeability compared to water, and the concentrate is used, thereby increasing the amount of active ingredient added to the jujube seed and improving skin penetration rate to obtain an excellent skin improvement effect.

Jujube seed, ethanol, soap

Description

Soap containing the extracts of Zizyphus jujube seed and manufacturing method of it}

The present invention relates to a soap using a jujube seed extract and a method for producing the same, and more particularly, extracts the active ingredient from ethanol discarded without being used, using ethanol to reuse the discarded jujube seed, effective of jujube seed The present invention relates to a soap using a jujube seed extract and a method for producing the same, which can obtain a skin improving effect due to an ingredient and a cell membrane permeability improving effect due to the use of ethanol.

Jujube contains about 0.55 to 3.5% of organic acids such as vitamins, amino acids, malic acid, succinic acid, etc., and contains various vitamins A, B2, C, and P.

In oriental medicine, the dried jujube is known to be effective in treating various diseases such as not only warming the body but also nourishing the skin, bleeding, stabilizing the mind, detoxifying various poisons, and anti-allergic effects. In particular, jujube is known to contain a large amount of vitamin C and trace elements, so that when taken regularly, the skin is smooth and clean, the color is healthy, and the face is beautiful.

Such jujube is usually used in porridge, tea, rice cake, medicine rice, etc. by using the flesh from which seeds are removed, or recently, it is used in various processing forms such as jujube drink, liquid jujube tea, granulated jujube tea, jujube fructose, jujube syrup.

In addition, a number of techniques using jujube extract have been disclosed in connection with the use of the jujube. For example, Korean Patent No. 0583051 “Cinnamon and Jujube Extract for Cancer Treatment and Prevention” includes cancers including cinnamon and jujube extract. The present invention relates to pharmaceutical compositions and functional foods for treatment and prophylaxis, and discloses the effect of inhibiting cell death and cancer cell migration by each component and the increase of cancer cell growth inhibitory effect by mixing the components. No. 99-0039367 discloses the composition of the antihypertensive agent or health food for lowering blood pressure, including the jujube extract, in the “composition for lowering blood pressure comprising jujube extract”, Republic of Korea Patent Publication No. 10-2008-0080719 In the lake, jujube was centrifuged with distilled water, extracted with ethanol, and washed with soap. Methods of making are disclosed.

As described above, processed foods using various jujubes and various prior arts mainly use the pulp or use the whole, and do not present a method of using the seed discarded after being used in processed foods.

On the other hand, the Republic of Korea Patent No. 10-0801902 registered and filed by the applicant of the present invention to extract the active ingredient from the jujube seed using water or methanol has an anti-cancer, antioxidant action, the technology to use it as a food or medicine is disclosed have.

However, the above-described prior art is limited to the use of ingested products such as foods and medicines, and is limited to extraction using methanol and distilled water. In particular, methanol has excellent cell membrane permeability and can be introduced through the skin. It has been reported that fatal symptoms have occurred through this pathway, and the symptoms are similar to those caused by "inhalation." These methanol causes mild skin itching, dryness and cracking on contact, cause minor or mild eye irritation, blindness or death upon ingestion, and ingestion of methanol below lethal doses may cause vomiting, headache, abdominal pain, vomiting and cloudy May cause symptoms that are sensitive to vision and light, and may cause nervous system intoxication, brain disorders, blindness and blindness when exposed to methanol (chronic) exposure through inhalation or skin exposure. On the other hand, water has a problem of low cell membrane permeability compared to methanol.

The present invention obtains the extract using ethanol excellent in cell membrane permeability compared to water while reusing the jujube seed discarded in processed foods using conventional jujube, and using the concentrate, increase the amount of active ingredient added to the jujube seed and improve skin penetration rate The purpose is to achieve excellent skin improvement effect.

A feature of the present invention for achieving the above object is a soap, characterized in that the addition of 0.5-10% by weight of the total weight of the jujube extract extract concentrated under reduced pressure concentrated ethanol extract of jujube seed.

And another feature of the present invention is to add 5-15 times 99% ethanol to the jujube seed powder sample and repeated 1-5 times extraction for 5-10 hours in a water bath equipped with a reflux cooler and then filtered the filtrate A jujube seed extract concentrate is prepared by concentrating under reduced pressure, and the soap production method includes a soap manufacturing step of preparing a soap by adding the above jujube seed extract concentrate to 0.5-10% by weight of the total weight.

In the present invention, since the jujube seed is discarded without being used in processed foods manufactured with ordinary food, it is possible to obtain an income improvement effect along with a resource recycling effect.

In addition, since the extract is obtained using ethanol having excellent cell membrane permeability compared to water, and the concentrate is used, the amount of active ingredient added to the jujube seed included in the soap is increased compared to the case of using the non-concentrate, and thus the ethanol having excellent cell membrane permeability is used. Compared with the conventional extract using water, the skin penetration rate is improved, and the skin improvement effect can be obtained.

In addition, methanol having excellent cell membrane permeability causes a problem on the skin, but the above-mentioned extraction using ethanol does not cause any problem on the skin, and it is helpful for skin improvement because of its disinfection effect and excellent cell membrane permeability.

In addition, the above jujube seed extract has anti-cancer, anti-oxidant effect, thereby improving the skin by such an active ingredient.

In addition, ethanol increases the chemical reaction rate of the fatty acid and potassium hydroxide has the effect of reducing the manufacturing time during the production of soap.

Hereinafter, an embodiment of the present invention will be described in more detail.

1. Jujube Seed Extract Extract

Jujube ( Ziziphus jujube ) used in this experiment was cultivated in Gyeongsan, Gyeongsangbuk-do.

After adding 10 times 99% ethanol to the jujube flesh and jujube seed powder samples, the extract was filtered three times for 8 hours in a water bath equipped with a reflux condenser and filtered. The filtrates were concentrated under reduced pressure and used as an ethanol extract.

These samples were used for the experiment while stored at -30 ℃.

2. soap manufacturer

The soap manufacturing method is known as the MP (melt and pour) method, CP (cold process) method, HP (hot process) method, Rebatch (Rebacthing) method, etc., any of these methods may be used, jujube The amount of seed extract concentrate added is 5g-100g when making 1kg soap.

Look at one embodiment of a soap manufacturing method.

Step 1: Fatty acid oils used in soaps are already known, and according to the purpose to be prepared, the fatty acid fats shown in Table 1 are selected and used.

For example, 40-60% olive oil and 60-40% coconut oil may be used in a weight ratio of 100%.

Second step: 20-35% caustic soda and 65-80% of purified water are mixed by weight, followed by a step of cooling the mixture at room temperature and maintaining it at 25-43 ° C, and adding caustic soda to purified water.

Third step: While maintaining the fatty acid, stirring while heating over low heat until 40-50 ℃.

Step 4: The stirred fatty acid oil is added at 40-70% by weight, and the mixture of caustic soda and purified water is added at 30-60% by weight, but the temperature is maintained at 40-50 ° C. and slowly poured while stirring. Go through the steps

Step 5: Injecting 2-6% by weight of the jujube seed extract compared to the agitated mixture to the agitated mixture.

The soap solution stirred in this way is put into a mold and cured.

Looking at the second embodiment of the soap manufacturing method, 1kg soap base, 15g olive oil, 15g glycerin, 10g essential oil, 30g jujube seed extract concentrate, anhydrous ethanol is prepared.

Cut a small amount of soap base into a beaker and melt it using a hot plate (microwave), then stir well by mixing the weighed additives (olive oil, glycerin, jujube seed extract concentrate) with the soap base.

When the temperature of the well-mixed soap liquid is lowered to 45-50 ℃, add essential oil and stir, and then pour the soap liquid into the mold while maintaining the temperature of the soap liquid at 45-50 degrees.

After the soap is poured into the soap mold, if the surface bubbles, sprinkle a little ethanol.

When soap solidifies at room temperature, remove the soap from the mold.

The soap thus produced has a transparent brown color as shown in FIG. 1.

3. Efficacy test of jujube seed extract

(1) As the jujube seed extract concentrate, the concentrated ethanol extract was used.

(2) reagent

Benzo (a) pyrene, 4-nitroquinoline-1-oxide, nicotinamide adenine nucleotide phosphoric acid (NADP), reduced nicotinamide adenine dinucleotide phosphoric acid (NADPH), D- D-glucose-6-phosphate, sodium dithionite, acrylamide, TEMED (N, N, N ', N'-tetra methyl ethylene diamine), bovine serum albumin (bovine serum albumin), pyrogallol, glutathione (reduced), SDS, etc. from Sigma (USA), D-biotin and L-histidine, etc., Junsei Chemical Co. (Japan) product was used. And N-methyl-N-nitro-N-nitroso-guanidine (MNNG) from Aldrich Chemical Co. (USA), aroclor 1254 from Chem. The immobilon PVDF membrane, ECL detection kit was purchased from Lite (German) from NEN ^TM Life Sciences (USA), and the polyclonal antibody Anti-rat CYP1A1 (goat) was purchased from DAIICHI Pure Chemicals (Japan). DMEM, FBS, and penicillin G / streptomycin were purchased from Gibco (USA), and other reagents were used as express products.

(3) antioxidant experiment

DPPH (1.1-diphenyl-2-picrylhydrazyl) radical scavenging ability of the extract was measured by Yoshikawa et al.

Samples and 0.2 mM DPPH solution were added to 0.1 M acetate buffer (pH 5.5), shaken for 10 seconds, and reacted for 30 minutes at 37 ° C. The reaction solution was then 517 nm using a spectrophotometer (Spectronic GENESYS 5, MILTON ROY). Absorbance was measured at, and the results are shown in FIG. 1.

(4) antimutagenic experiment

The antimutagenic effect of the sample extract was examined using the Ames mutagenicity test. The strains used were Salmonella typhimurium TA98 and TA100 from Dr. Bruce N. Ames of the University of California, Berkeley. Genetic strains such as deep rough (rfa) mutation, uvrB mutation, and R-factor were used as experimental strains. The genotypes of these strains are shown in Table 2.

Salmonella typhimurium TA98 and TA100 strains were routinely identified as genotypes such as histidine requirements, deep rough ( rfa ) mutations, uvrB mutations, and R fator .

Mutogens and carcinogens were examined. Benzo (a) pyrene (B (a) P) was used as an indirect mutagens in DMSO. Direct mutagens were tested by dissolving N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitroquinoline-1 -oxide (4-NQO) in distilled water and DMSO.

S9 mixture is a liver microsomal enzyme mixture prepared according to the method of Maron and Ames in order to activate the indirect mutagenesis factor using the metabolic activity system of the animal. In order to induce microsomal enzyme from Sprague-Dawley male rats weighing about 200 g from Korean experimental animals, aroclor 1254, a polychlorinated biphenyl (PCB) mixture, was diluted with corn oil and injected once intraperitoneally (500 mg / kg). After 5 days the livers were removed. The extracted liver was washed with ice-cooled 0.15M KCl solution in sterile state, and then added with 3ml of 0.15M KCl per gram of liver, chopped with sterile scissors and homogenized with a homogenizer. This was centrifuged at 9,000 × g and 4 ° C. for 10 minutes to obtain a S9 fraction as a supernatant. 1.5 mL of this fraction was dispensed into cryogenic vials and rapidly frozen and stored at -80 ° C. The S9 mixture contains S9 fraction (10%) with MgCl-KCl salts (2%), 1M glucose-6-phosphate (0.5%), 1M NADP (4%), 0.2M phosphate buffer (pH7.4) and sterile water. The mixture was prepared.

The mutagenicity and antimutagenicity of samples and mutagen were examined by preincubation mutagenicity test using methods such as Maron, Ames and Matsushima.

In the preincubation mutagenicity test, 0.5 ml of S9 mixture (indirect mutation) or 0.5 ml of phosphate buffer (direct mutation), 0.1 ml of strain incubated for 12 hours, 50 µl of sample, and 50 µl of mutagen were added to the cap tube containing ice bath. After mixing lightly and pre-incubated at 37 ° C. for 30 minutes, 2 ml of top agar at 45 ° C. was added to each tube and mixed for 3 seconds. This was plated on a minimal glucose agar plate and incubated at 37 ° C. for 48 hours to count the number of revertant colonies.

Meanwhile, the concentrations of the sample and mutagen used in the experiment were determined through preliminary experiments (dose response and toxicity experiment).

Inhibition rate was calculated by Equation 1 below.

Inhibition rate (%) = [(a-b) / (a-c)] × 100

a: number of return mutations induced by the mutagen

b: number of return mutations when the sample is processed

c: Natural return mutations without mutagen and sample

A date palm seed extract the S almonella typhimurium TA100 N-methyl- N`-nitro-nitroso-gaunidine represents an antimutagenic effect using in Table 2, wherein by using a 4-nitroquinoline1-oxide of jujube seed extract in S typhimurium TA100 mutations almonella The effect is shown in Table 3, and the indirect mutation effect using benzo (a) pyrene on Salmonella typhimurium TA98 in jujube ethanol extract is shown in Table 4, and the mutation inhibition effect of jujube was detected by Ames test. The mutagenicity was not found, and mutagenic effects were observed for the direct mutant N-methyl-N'-nitro-N-nitrosoguanidine and 4-nitro-quinoline-1-oxide and the indirect benzo (a) pyrene.

     Treatment Revertants / plate Inhibition
rate (%) Spontaneous 111 MNNG 840.5  MNNG + H ₂ O ext.              10% 327.5 37.4               5% 518 44.2             2.5% 567.5 70.3

In the above, MNNG (N-methyl-N'-nitro-nitrosoguanidine) 0.2 µg / plate.

     Treatment Revertants / plate Inhibition
rate (%) Spontaneous 111 MNNG 1156  MNNG + H ₂ O ext.              10% 342 77.9               5% 523.5 60.5             2.5% 703.5 43.3

In the above, 4-NQO (4-nitroquinoline-1-oxide) is 0.1 µg / plate.

     Treatment Revertants / plate Inhibition
rate (%) Spontaneous 38.5 MNNG 670  MNNG + H ₂ O ext.              10% 150.5 82.3               5% 298.5 58.8             2.5% 593.5 12.1

In the above, Benzo (a) Pyrene 10µg / plate.

(5) measurement of cancer cell proliferation inhibitory effect

The cell lines used in the experiments were human hepatoma cells Hep G2 (hepatoma cell, human) and colon cancer cell HT-29 (colon carcinoma cell, human).

HT-29 cells were cultured with 10% FBS (10% heat inactivated fetal bovine serum) and 1% antibiotics in RPMI-1640 medium, and HepG2 cells were cultured with 10% FBS and 1% antibiotics in MEM medium. . The cells were grown in monolayers on a disposable cell culture plate (100㎠, corning), and the culture temperature and CO ₂ concentration were maintained at 37 ° C. and 5% CO ₂ for adaptation and cultured every two to three days. Used.

The MTT assay is a colorimetric assay that measures the growth and differentiation of cells based on the production of dark blue formazan by MTT, a yellow water-soluble substance of dehydrogenase in living mitochondria.

The inhibitory effect of jujube extracts on the hepatoma cell Hep G2 (hepatoma cell, human) and colon cancer cell HT-29 (colon carcinoma cell, human) was investigated by MTT assay according to Green et al.

First, remove the medium from the cultured cells, mix lightly with PBS, remove PBS again, and incubate for 5 minutes at 37 ° C with 0.25% trypsin-EDTA to observe whether the cells were completely separated from the bottom of the culture dish. After the addition of the culture medium and mixed well, the cell number was adjusted to 1 × 10 ⁵ cells / ㎖ was used in the experiment. Samples were dissolved in DMSO at the appropriate concentration and filtered through a 0.22㎛ membrane filter. Each cell prepared in a 96-well plate was dispensed 198 μl and incubated for 24 hours at 37 ℃, 5% CO ₂ . 2 μl of each concentration sample was added thereto and incubated for 48 hours. Then, 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT, 0.5 mg / ml) In addition, MTT was reduced by enzymatic action of viable cancer cells. After 4 hours, the formazan formed on the bottom of the well was carefully treated to remove the supernatant with an aspirator. 200 μl of DMSO was added to each well to dissolve formazan and the absorbance was measured at 540 nm using a microplate reader.

The absorbance represents the amount of MTT decomposed into cells by formazan. Therefore, the absorbance is proportional to the number of viable cells in each well, and cytotoxicity is calculated as follows.

The anticancer effect of colon cancer cell HT29 using jujube ethanol extract of the present invention is shown in Figure 2, and the anticancer effect of hepatic cancer cell HepG2 using jujube ethanol extract is shown in Figure 3.

Jujube ethanol extract has a high cancer cell proliferation inhibitory effect can be confirmed through 2 and 3.

1 is a view showing a soap according to the present invention

2 is a view showing the antioxidant effect of jujube seed extract according to the present invention

3 to 4 is a view showing the anticancer effect of jujube seed extract according to the present invention

Claims (2)

delete After adding 5-15 times 99% ethanol to the jujube seed powder sample, the extract was repeated 1-5 times for 5-10 hours in a water bath equipped with a reflux condenser and filtered, and the filtrate was concentrated under reduced pressure. Preparation steps, Soap manufacturing method comprising a soap manufacturing step of adding the above jujube seed extract concentrate to 0.5-10% by weight of the total weight to prepare a soap.

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