KR101796423B1 - Composition and ladies genital area cleanser composition comprising composition for inducing Antiviral and antibacterial cause vaginitis and method for manufacturing the same - Google Patents

Abstract

The present invention relates to methods of producing an antiviral and antibacterial composition capable of suppressing virus and pathogen causing vaginitis, and a feminine wash composition comprising the same. Specifically, the method of producing a feminine wash composition comprises: a drying step (10) for harvesting and drying Coptis chinensis, Magnolia Kobus bark, Centella asiatica, and Glycyrrhiza Glabra, all of which are natural substances; an extraction step (20) for producing an extract of each Coptis chinensis, Magnolia Kobus bark, Centella asiatica, and Glycyrrhiza Glabra; a filtering step (30) for filtering the extracts; a sterilization step (40) for sterilizing the extracts at a high temperature; a concentration step (50) for concentrating the extracts; a concentration adjustment step (60) for diluting the extracts so as to make concentrations uniform; a mixing step (70) for mixing the extracts to produce a composition solution; and a dilution step (80) for diluting the composition solution before use.

Description

[0001] The present invention relates to an antiviral and antimicrobial composition capable of inhibiting viral infections and infectious microbes causing vaginitis, and a method for producing a female cleanser composition comprising the same. [0002]

The present invention relates to a natural female cleanser composition comprising a natural substance, such as Rhodiola, Magnolia bark, Centella asiatica, Licorice extract, Dipotassium Glycyrrhizinate, which can inhibit specific viruses and infectious bacteria causing vaginitis The present invention relates to an antiviral and antimicrobial composition harmless to the human body, and a method for manufacturing a female detergent composition comprising the same.

Female genitourinary system diseases include ovarian cancer, uterine leiomyoma, uterine polyp, uterine cervix, uterine and vaginal prolapse, uterine cancer, choriocarcinoma, uterine cervix erosion, cervical dysplasia, cervical cancer, vulvar vaginitis, vaginal yeast infection, bacterial vaginitis, These diseases are relatively common, and most women experience at least one disease in their lifetime. These are the most common diseases, such as vulvar and vaginal cancer, sexually transmitted diseases (gonorrhea, chlamydial pelvic inflammation, syphilis, trichomoniasis, genital herpes, vulvar warts and genital warts).

In female genitourinary system, lactic acid bacterium, which maintain weak pH of 4.5-5.5 in urinary genital system, plays a role in preventing the propagation of harmful bacteria. However, in addition to physical impairment, hormone imbalance, unsterile genitourinary environment, bacterial disease due to sex, The female genitourinary system-related diseases are likely to occur.

The vagina and vulva are directly exposed to the external environment due to the functional nature of the female genitourinary system and provide a conditional environment in which various bacterial populations such as transference, temperature and humidity are cultivated to cause bacterial infections and bacterial vaginosis.

There are two pathogenic viruses and pathogens associated with vaginitis. Among them, HSV (Herpes Simplex Virus) has two types of HSV-1 and HSV-2. HSV-1 is a virus that causes herpes zoster HSV-2 is a genital herpesvirus that is transmitted to newborns when sexual contact and maternity are delivered. It is a viral herpesvirus that causes swelling around the penis, buttock, and vaginal vagina when tired. Virus.

Human Immunodeficiency Virus (HIV) is a virus that causes AIDS and is transmitted to people through blood transfusions or sexual contact. The HIV-1 virus, commonly referred to as HIV, and HIV-2, the second form of the human immune virus, are both closely linked in that they cause AIDS.

Gonorrhea gonorrhea ( Neisseria gonorrhoeae ) refers to a disease caused by bacteria. Infections can occur in women's uterus, fallopian tube, vagina, cervix, and anus, but they can cause infertility if left untreated.

Mycoplasma Hominis is a bacterium that is associated with non-gonococcal urethritis, prostate urethritis, postpartum fever, endometritis, and cervicitis. Infections do not cause any major symptoms, and excessive proliferation can cause serious inflammatory diseases, leading to infertility and abortion.

Ureaplasma spp. Is commonly found in women with sexual intercourse. There is no such symptom, but excessive proliferation causes serious inflammation and complications.

Candida albicans , a fungal species, is found in many parts of the body such as the gastrointestinal tract, vagina, urethra, skin, nails, etc. It causes candida vaginitis in pregnant women and others.

Escherichia coli Is the most common causative organism of urinary tract infections. Women have a higher incidence of urinary tract infections than men, and 40-50% of all women report urinary tract infections at one time.

Accordingly, the present invention provides a composition for inhibiting the growth of viruses, bacteria and fungi capable of generating vaginitis, and as a prior art of the Korean Patent Office related to a female cleansing agent using natural substances, Patent No. 10-2006-0036290 discloses a composition for women's cleanser comprising bamboo salt and herbal extracts containing yugenmei, gosam, gold, ginkgo leaf, licorice and chamomile in a female cleansing composition comprising bamboo salt and herbal extracts And 2) Patent Application No. 101125758 entitled " Composition for cleansing composition for women's local cleansing " discloses a composition for local cleansing of women consisting of gosam, lobster, guinea fowl, peony root, And 3) Patent No. 101168225 'Combined herbal medicine fermented extract and a female cleansing agent containing the same Discloses a composition for a women's cleansing composition of a complex fermentation extract of green tea, astragalus, Angelica keiskei, radish, and persimmon, and 4) in a 'mild female cleanser composition' of Patent Publication No. 10-2014-0092635, acyl lactylate A composition using a plant extract containing an anionic surfactant and saponin is disclosed, and 5) a antibacterial composition having an effect on prevention or treatment of vaginitis and a female cleaner composition containing the same is disclosed in Korean Patent No. 101345709 And 6) a composition of a female cleansing composition comprising a natural raw material extract of Porphyry, Artemisia princeps, and Artemisia flowers, a composition thereof, and a composition thereof, Use method and manufacturing method ', a female cleanser composition prepared by extracting a pupa, an artificial horse mugwort, And 7) Patent Document 10-2015-0111663, "Natural Women's Cleaner", 8) Patent No. 101651530, "Method of Manufacturing Female Cleaner Using Herbal Extract" In the present invention, antiviral and antimicrobial compositions capable of inhibiting specific viruses and infectious microorganisms causing vaginitis by using dipotassium glycyrrhizate, which is an active ingredient of Rhodiola, Magnolia bark, Centella asiatica, licorice extract and licorice extract, To a method for manufacturing a female cleaner composition.

1. Korean Patent Publication No. 10-2006-0036290 (2006.04.28) 2. Korean Registered Patent No. 101125758 (2012.03.05) 3. Korean Patent No. 101168225 (July 18, 2012) 4. Korean Patent Publication No. 10-2014-0092635 (2013.01.16) 5. Korean Patent No. 101345709 (Dec. 20, 2013) 6. Korean Patent No. 101432996 (2014.08.18) 7. Korean Patent Publication No. 10-2015-0111663 (June 10, 2015) 8. Korean Patent No. 101651530 (Aug. 26, 2016)

1. Cor. J. Food Nutr. 18: 81-87.2005 2. Cor. J. Herbology 2014; 29 (5): 83-90 3. Cor. J. Microbiol. Biotechnol. 37, No. 1, 42 ?? 8 (2009) 4. J. Soc. Cosmed. Scientists Korea Vol. 39, No. 1, March 2013,1-8

Women's cleanliness moisturizers include chemical compositions and natural compositions. If they are frequently cleaned with artificial detergents and the like, the natural balance of the function of the midnight function may be broken, resulting in overgrowth of harmful bacteria and yeast infection. Therefore, it is preferable to use a natural substance because it is a cleanliness moisturizer composition used in a sensitive area, but it is not easy to form a combination of natural substances having a high effect.

It is an object of the present invention to find an excellent natural extract composition as a female cleanser and to apply it to product development.

Another object of the present invention is to provide a method for manufacturing a safe female composition for a female Y zone for the prevention and amelioration of an inflammatory disease by enhancing the activity of microorganisms of vaginal use in women and inhibiting the proliferation of certain harmful microorganisms.

In addition, another object of the present invention is to provide a method for producing Candida albicans and Escherichia coli Pies and specific harmful pathogenic virus, HSV, HIV and gonorrhea Hospital gyunin (Neisseria gonorrhoeae ), Mycoplasma hominis and Ureaplasma spp . The present invention provides a method for manufacturing a female cleaner composition.

Another object of the present invention is to provide a method for preparing a female detergent composition comprising an antiviral and an antimicrobial composition harmless to the human body while suppressing desired pathogenic viruses and pathogens using natural extractives.

It is an object of the present invention to provide a method of preparing a female detergent composition which is excellent as a female cleansing agent, extracts a natural extract composition, enhances the effect of inhibiting pathogenic viruses and pathogens, and is stable in the human body.

Therefore, according to an aspect of the present invention for achieving the above-described object,

A drying step (10) for harvesting and then drying the natural substances such as Rhododendron, Magnolia bark, Cinnabar, and licorice;

An extracting step (20) of extracting 1 kg of Rhodiola prepared in the drying step, 10 kg of water, 1 kg of magnolia seedlings, 1 kg of Ciliaceae and 1 kg of licorice at 90 to 100 ° C for 10 to 24 hours until the total weight reaches about 7 Kg;

The extraction solvent may be a polar solvent selected from the group consisting of water and a mixed solvent thereof, and may have a mixing ratio (kg / L) of about 10: 1 to 1:10, preferably 1: 1 to 1: 3 Can be used.

A filtration step (30) of filtering the extract liquid extracted in the extraction step with a cartridge filter (10 microns);

A sterilization step (40) for pasteurizing the filtered extract at 95 ° C in the filtration step;

A concentrating step (50) of concentrating the extract sterilized in the sterilization step to a concentration of 10-20 brix using a freeze dryer or a rotary vacuum concentrator;

A concentration correction step (60) of adding the extracts concentrated to be 10 to 20 brix in the concentration step to dilute the extract so as to have a concentration of 3 to 6 brix, thereby making the concentration of the extract uniform;

10 to 40 parts by weight of the extract of Magnolia bark, 10 to 40 parts by weight of the centipede extract, 10 to 40 parts by weight of the licorice extract, 10 to 40 parts by weight of the extract of DPG (Dipotassium Glycyrrhizinate) in an amount of 0.2 to 10 parts by weight to form a composition stock solution (70);

A diluting step (80) for diluting the mixed raw material solution and water at a weight ratio of 1: 5 to 1:20 in order to use the raw material of the composition prepared in the mixing step as an actual composition;

.

The present invention relates to a composition of Dipotassium Glycyrrhizinate which is an active ingredient of Rhodiola, Magnolia bark, Cinnabar, licorice extract and licorice extract, and a natural female clean product containing the same. According to the present invention, by inhibiting the proliferation of certain harmful microorganisms, Candida albicans and Escherichia coli And HSV, HIV, and Gynecologic Gynecology, N eisseria gonorrhoeae ), M ycoplasma hominis and U reaplasma spp. And particularly, it has an effect of providing a stable female cleaner composition in the human body.

BRIEF DESCRIPTION OF THE DRAWINGS FIG.
Figure 2 is a graphical representation of the composition of the Neisseria Gonorrhoeae is a photograph of inhibition.
Figure 3 is a photograph of the fungus Candida It is a photograph of growth inhibition ability of albicans and bacterium Escherichia coli .
Fig. 4 is a graph showing the effect of the fungus Candida Disc diffusion methods of albicans and Escherichia coli bacteria.
Figure 5 is a photograph of the fungus Candida < RTI ID = 0.0 > It is a graph of the fluorescence value of inhibition ability of albicans after 0 to 18 hours of reaction.
Fig. 6 is a photograph of the fungus Candida Plate photographs of inhibition of albicans after 0 to 6 hours of reaction.

Hereinafter, the present invention will be described in detail. In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to be exemplary, self-explanatory, allowing for equivalent explanations of the present invention.

In the present specification, the term " inhibitory force " means inhibiting the growth or proliferation of a substance to be treated or killing microorganisms.

Dipotassium glycyrrhizinate, an active ingredient in the extracts of Rhododendron, Magnolia bark, Clove, Licorice extract and licorice extract, was found to be effective against pathogenic viruses and pathogens in various natural resources. It was confirmed that there is a pathogen inhibiting ability, and an excellent natural extract composition was discovered as a female cleansing agent using a natural substance composition and applied to product development.

C optis chinensis is the origin of China and has been used in vomiting for diarrhea and poisoning in the private sector and is known to have fever and detoxification (Kor. J. Food Nutr. 18: 81-87.2005). Especially, antimicrobial agents such as berberine palmatine, coptisine, magnoflorine, epiberberine, berbestine, worenine and ferulic acid have been reported to be effective for anti-inflammation (Kor J. Herbology 2014; 29 (5): 83-90) (Kor. J. Microbiol. Biotechnol. Vol. 37, No. 1, 428 (2009)).

Magnolia bark extract ( Magnolia kobus bark) is reported to have antioxidant, anti-inflammatory, anti-cancer, anti-thrombotic, antibacterial, antiallergic and antifungal effects. Purified Magnolia bark extract contains a very high content of two highly chemically related substances, honokiol and magnolol, which are being studied in a number of ways, including antioxidant and antiinflammatory actions, Staphylococcus aureus, Staphylococcus hemolyticus, and Bacillus diphtheriae.

Centella asiatica is extracted from the leaves and roots of Gotu Kola trees grown in India, Sri Lanka and South Africa. It has been used for skin in India since ancient times. The main ingredients are asiaticoside, madecassic, and asiatic acid. Among the constituents, matecaside is contained in a small amount as an extract. However, it is used as a therapeutic agent for herbal medicine by highly refining only this ingredient. The efficacy has been widely used as a folk remedy for managing health and beauty for many years. It has a high antioxidant effect and is used not only as a herb but also has an antibacterial effect which inhibits bacillus causing skin tumors, It contains phytosterol and glycotothelin which are highly anti-inflammatory and sedative. It helps to calm red skin. It helps the collagen production of the dermis to help improve elasticity and pores.

Glycyrrhiza Glabra has been used only as a whitening agent, but nowadays, natural antioxidants (J. Soc. Cosmed. Scientists Korea Vol. 39, No.1, March 2013,1-8), antimicrobial activity, natural preservatives And many other studies have been conducted and used.

In particular, DIPOTASSIUM GLYCYRRHIZINATE is a dipotassium salt of glycyrrhizic acid, which is a kind of natural component extracted from licorice. Dipotassium glycyrrhizate, also known as licorice acid, has been reported to be effective for regulating sebum, moisturizing effect, skin whitening, and anti-inflammation as well as restoring the damage of epidermal barrier of skin.

An object of the present invention is to find a natural substance composition for inhibiting pathogenic viruses and pathogens as a female cleansing agent to thereby prepare an antiviral and antimicrobial composition and a female cleansing composition comprising the same.

1 shows a method of manufacturing a female cleaner composition according to an embodiment of the present invention.

Referring to FIG. 1, the method for preparing the female cleaner composition comprises a drying step (10); Extraction step (20); Filtration step (30); A sterilization step 40; Concentration step 50; A density correction step (60); Mixing step 70; And a dilution step (80).

Hereinafter, a method for manufacturing a female cleaner composition according to the present invention will be described step by step.

Hereinafter, the configuration and operation of the present invention will be described in more detail with reference to preferred embodiments of the present invention. It is to be understood, however, that the same is by way of illustration and example only and is not to be construed in a limiting sense.

<Female Cleaner Manufacturing Method>

(10) Drying step

It harvests natural materials such as chrysanthemum, magnolia bark, cinnabar, and licorice.

(20) Extraction step

10 kg of water is boiled for 1 to 24 hours at 90 to 100 ° C until the total weight reaches about 7 Kg, 1 kg of Rhodiola prepared in the drying step, 1 kg of magnolia tree, 1 kg of lupine and 1 kg of licorice are boiled for 10 to 24 hours.

The extraction solvent may be a polar solvent selected from the group consisting of water and a mixed solvent thereof, and may have a mixing ratio (kg / L) of about 10: 1 to 1:10, preferably 1: 1 to 1: 3 Can be used.

(30) Filtration step

Filter the extract from the extraction step with a cartridge filter (10 microns).

(40) Sterilization step

In the filtration step, the filtered extract is sterilized at 95 ° C.

(50) Concentration step

The extract solution sterilized in the sterilization step is concentrated to 10-20 brix using a freeze dryer or rotary vacuum concentrator.

Since the content of natural substances per unit and the yield in extraction process are different from each other, it is necessary to quantify each natural substance once it has been concentrated to a high concentration, and to prepare an extract solution having the same concentration in the following concentration correction step .

(60) Concentration correction step

In the concentration step, the extracts concentrated to 10 ~ 20brix are diluted to 3 ~ 6 brix with distilled water to make the concentration of each extract uniform.

(70) mixing step

10 to 40 parts by weight of Magnolia bark extract, 10 to 40 parts by weight of Centella asiatica extract, 10 to 40 parts by weight of licorice extract, And 0.2 to 10 parts by weight of DPG (Dipotassium Glycyrrhizinate) to prepare a stock solution of the antimicrobial composition.

DPG (Dipotassium Glycyrrhizinate) was purchased from Jiangsu Tiansheng Pharmaceutical Co., Ltd .; (68797-35-3).

The extracts may be mixed at a rotating speed of 100 to 500 rpm for 1 to 6 hours.

(80) Dilution step

The mixed stock solution of antimicrobial composition and water is diluted at a weight ratio of 1: 5 to 1:20 in order to use the stock solution prepared in the mixing step as an actual female cleaner composition.

The female cleansing composition of the present invention can be formulated into a liquid preparation such as a solvent, a suspension, an emulsion, a tincture, a syrup, a spray or an extract.

The composition prepared by the above-mentioned method for producing a female cleanser is used to treat HSV, HIV which is a peculiar female specific harmful virulent virus, Neisseria gonorrhoeae ), Mycoplasma hominis and Ureaplasma spp. , HSV1 inhibition test in Experimental Example 1, HIV1 inhibition test in Experimental Example 2, Neisseria in Experimental Example 3, gonorrhoeae Inhibition test and Mycoplasma hominis & Ureaplasma spp. Inhibitory test, and in Experimental Example 5, the fungus Candida albicans and bacteria Escherichia E. coli . Especially, since it is applied to sensitive areas of women, stability is important. Thus, in Experimental Example 6, a primary human skin stimulation test was performed to confirm whether or not the human skin was stable. , A buoyancy test was carried out to confirm that it has resistance to fungi.

Experimental Example 1 HSV1 inhibitory activity MTT assay

MTT assay was performed to confirm that HSV1 was inhibited by this composition.

Test viruses and host cells

Herpes simplex virus type 1 (HSV1) strain F (ATCC VR-733) and host cell Vero cell (ATCC CCL-8) were purchased from American Type Culture Center (ATCC)

Virus culture

For virus multiplication and titration, the virus-infected cells were used after incubation for 3 days in a 37 ° C CO ₂ incubator.

Experimental Method

For inactivation testing, Vero cells were placed in 96-well plates three days prior to testing and incubated in a 37 ° C CO ₂ incubator to allow complete cell monolayer formation on the day of testing.

To determine the difference between the concentration of the virus and the composition of the mixture

   (1) 50% Composition: 0.1 ml of virus, 0.5 ml of the stock solution (50%) and 0.4 ml of distilled water

   (2) 90% composition: 0.1 ml of virus and 0.9 ml (90%) of the composition stock solution

   (3) Control: 0.1 ml of virus and 0.9 ml of distilled water

   (4) 0.1 ml of mock: MEM / 2% FBS and 0.9 ml of the composition solution

Each experimental solution was made.

The test solution was 10 ⁰ ~ 10 ^-6 mg / ml during the 37 ℃ CO ₂ incubator for 1 hr to be treated, removing the supernatant and then washing, MEM / 2% per each well in MEM / 2% FBS The cells were incubated in a 37 ° C CO ₂ incubator for 3 days. Then, the culture supernatant was removed, and the virus titer and the cytotoxic concentration were determined by MTT staining to confirm the degree of virus inhibition.

result

As can be seen in Table 1, when 50% solution and 90% solution were reacted for 2.5 minutes or more, the reduction rate of 99.8% and 99.9% or more of HSV1 was confirmed.

Experimental Example 2 HIV1 inhibitory test

MTT assay was performed to confirm that HIV1 was inhibited by this composition.

Test viruses and host cells

The HIV-1 (human immunodeficiency virus type 1) strain IIIB was purchased from the National Institue of Biological Standards and Controls (NIBSC) based on the "MRC AIDS Reagent Project", and the MT4 cells used as host cells were obtained from Tokyo Medical and Dental University Professor N. Yamamoto of the University of Tokyo.

Virus culture

Cells infected with the virus were incubated in a 37 ° C CO ₂ incubator for 5 days for viral growth and titration.

Experimental Method

For inactivation testing, MT4 cells were placed in 96-well plates 5 days before the test and incubated in a 37 ° C CO ₂ incubator to allow complete cell monolayer formation on the day of testing.

To determine the difference between the concentration of the virus and the composition of the mixture

   (1) 90% Composition: 0.1 ml of virus and 0.9 ml (90%) of composition stock solution

   (2) Control: 0.1 ml of virus and 0.9 ml of distilled water

   (3) mock: 0.1 ml of RPMI1640 / 10% FBS and 0.9 ml of the stock solution

Each experimental solution was made.

The test solution was 10 ⁰ ~ 10 ^-6 mg / ml; for a treatment for 1 hour at 37 ℃ CO ₂ incubator, and that, after removing the supernatant, and RPMI1640 / washed with 10% FBS, RPMI1640 / 10% per each well After incubation for 5 days in 37 ° C CO ₂ incubator, the culture supernatant was removed and the viral titers and cytotoxic concentrations were determined by MTT staining.

result

As can be seen in Table 2, when 90% solution was reacted for 2.5 minutes or more, the reduction rate of HIV1 of 98% or more was confirmed.

< Experimental Example 3 > Neisseria gonorrhoeae  Inhibition test

In order to confirm that Neisseria gonorrhoeae bacteria were inhibited by the present composition, the inhibition experiment was carried out by culturing in a medium.

Experimental Method

For the test method, prepare a strain by mixing N. gonorrhoeae strain with a standard turbidity of 0.5 and mix the stock solution with the stock solution (1 ml①, 2 ml②, 3 ml ③, control (bacterium without added composition ④)) After reacting at room temperature for 5 minutes , Inoculated on Thayer-Martin agar and incubated for 24 to 48 hours in a 5% CO ₂ incubator at 35 ° C.

result

The composition N. gonorrhoeae Confirmation of colony formation by Gram staining after mixed culture 24h culture 48h culture N.gonorrhoeae ① No growth No growth N.gonorrhoeae ② No growth No growth N.gonorrhoeae ③ No growth No growth N.gonorrhoeae ④ (control group) Growth Growth

Figure 2 shows the result of Gram staining on Thayer-Matrin agar.

In FIG ①②③ group as identified in two of N1, N2, N3 agar plate, and Table 3 is a Gram-negative ssanggugyun has not been observed (Gram negative diplococci), the composition on the Gram stain Neisseria gonorrhoeae and the inhibitory effect of gonorrhoeae .

< Experimental Example 4 > Mycoplasma hominis  Wow Ureaplasma spp . Inhibition test

By this composition, M ycoplasma hominis and U reaplasma spp. In order to confirm that the bacteria were inhibited, the inhibitory experiment was carried out on the medium.

Experimental Method

In the experimental method, the bacterial solution is mixed with the stock solution (1 ml ?, 2 ml ?, 3 ml?) For 5 minutes at room temperature, and then the reaction solution is dissolved in the R1 (reconstitution solution) medium. Divide 3 ml in R1 medium into R2 urea-arginine broth. At this time, vortexing is performed so that the lyophilized R2 pellet completely dissolves. Immediately dispense 55 ml of well mixed broth into 22 cups on the strip and drop 2 drops of sterilized mineral oil into each cup. After lidding, the strip and the remaining broth (R2) are incubated at 36 ° C for 24-48 hours.

No growth was observed after 48 hours of culture negative, and color change was observed after 48 hours as a result of positive culture, and it was confirmed by the identified strain, colony count, and antimicrobial susceptibility.

result

Composition and virus Mycoplasma hominis  & Ureaplasma spp . Mixed culture results 24h culture 48h culture Mycoplasma hominis &
Ureaplasma spp . No Growth No Growth Mycoplasma hominis &
Ureaplasma spp. No Growth No Growth Mycoplasma hominis &
Ureaplasma spp . No Growth No Growth

As shown in Table 4, there was no change in color in the experimental groups ①②③, and the composition was Mycoplasma hominis & Ureaplasma spp. The effect of inhibitory effect on.

<Experimental Example 5> Candida albicans  Wow  Escherichia coli  Inhibition test

In order to confirm that the fungus Candida albicans and the bacterium Escherichia coli were inhibited by the present composition, the inhibition experiment was carried out in the culture medium.

Experimental Method

Each strain of 1.5X10 ⁵ cfu / ml was spread on an agar plate and incubated at 37 ° C for 12-24 hours,

(1) After incubation at 37 ° C for 12-24 hours on plates spreading a mixture of 40% of the composition stock solution and 60% of each media and 100% of each medium alone, the fungus Candida The growth inhibition of albicans and Escherichia coli bacteria was observed. For the Candida albicans media, sabouraud-dextrose was used, Escherichia The media for coli was luriabertani medium.

(2) In addition, 50 μL of distilled water and 10% composition stock solution were loaded on Disc Paper, and then incubated in minimal medium at 37 ° C for 12 to 24 hours to determine whether a clear zone appeared. Disc diffusion method And it is confirmed through law.

(3) Fungicide Candida albicans was cultured on sabouraud-dextrose media and spread on a plate. To confirm the inhibition rate by time, the 10% composition stock solution was spread over 1 hour, 2 hours, 4 hours and 6 hours Growth inhibition rate was measured by fluorescence using alamar blue reagent, and the number of bacteria was checked with a plate by time.

result

As can be seen in Figure 3,

A plate cultured with Sabouraud-dextrose alone for the fungus Candida albicans and a mixture of 40% of the undiluted solution of 10% composition and 60% of sabouraud-dextrose in the experiment (1) As can be seen, growth inhibition against Candida albicans was confirmed.

In addition, cultured using only luriabertani medium for the bacteria Escherichia coli plate and the 10% composition solution 40% and luriabertani medium the plates bacteria Escherichia As can be seen in (b) of Figure 3 cultured in a mixed solution of 60% The growth inhibitory ability against E. coli was confirmed.

As shown in FIG. 4, when the strain is inhibited by the sample in the Disc diffusion method, a clear zone is formed around the paper disc, and a 50 μL composition solution for the fungus Candida albicans The clear zone was confirmed in the high concentration 40% composition solution and the clear zone of 1.05Cm in the bacterial Escherichia coli was confirmed, so that the 10% to 40% composition solution contained the fungus Candida albicans and the bacterium It was confirmed that the growth inhibitory effect of Escherichia coli was effective.

In the experiment (3), after the 10% composition stock solution was spread, the rate of growth inhibition by time was observed with a fluorescence value (FIG. 5) through the alamar blue reagent and a plate number (FIG. 6) And the efficacy was maintained for 6 hours.

<Experimental Example 6> Primary human skin stimulation test

In order to confirm whether the composition prepared according to the present invention is stable to the skin, a human skin primary stimulation test was conducted.

The subjects were 31 males or females aged between 18 and 60 who met the criteria for selection and exclusion of the subjects.

This test was conducted in accordance with the Human Body Application Test Guideline [Food and Drug Administration], Regulation 2015.08] and related regulations. The report is based on the MFDA, PCPC Guidelines, and the test results performed according to the standard work instructions (SOP) Which is the result of

Experimental Method

After the test area is wiped with alcohol, 16 μL of the stock solution is dropped into the Finn chamber, placed on the test site, and fixed with micro pore tape. After removing the patches, the test area was marked with a skin marker, and each test site was observed after 30 minutes and 24 hours. Observations were made 30 minutes and 24 hours after removal of the patch. Skin reactions were evaluated according to the following criteria, reflecting the Frosch & Kligman method. The test results are shown in Table 5.

result

 As shown in Table 5, the composition had no skin reaction or side effects, and was judged to be a substance of hypomodality category in human skin primary stimulation.

<Experimental Example 7> Flotation test

The buoyancy test was conducted on this composition against fungal Asperaillus nigers were performed in the following manner.

Experimental Method

Fungal Asperaillus nigers were diluted 1/10 of the composition stock solution and applied on a filter paper. The size of the solution was adjusted to 3 x 3 cm, placed on each bacterium, yeast and fungus growth medium, and each bacterium, yeast, After incubation for 4 weeks in an incubator at 29 ℃ and relative humidity of 80 ~ 90%, the fungus growth on the surface of each filter paper was observed for 1 week to 4 weeks in 1 week. In Table 6.

Fungal spores are Asperaillus nigers ATCC 9642 were used, and the fungal growth medium was a PDA (Potato Dextrose Agar) medium.

result

Buoyancy  Test result Sample classification unit result Test method Early CFU / ml 4.2X10 ⁵

CTFA Microbiological
Guidelines: 2007 Seven days CFU / ml &Lt; 10 (99.9% or more) After 14 days CFU / ml &Lt; 10 (99.9% or more) After 21 days CFU / ml &Lt; 10 (99.9% or more) After 28 days CFU / ml &Lt; 10 (99.9% or more)

As shown in Table 6, when it was judged according to the CTFA standard, it was found that the Asperaillus niger , which is a fungus type 1, was reduced by at least 99% within 7 days of inoculation,

In the method for manufacturing a female cleanser composition of the present invention, it is possible to provide an excellent natural extract composition as a female cleanser using a natural substance, improve the effect of inhibiting pathogenic viruses and pathogens, and provide a method for manufacturing a female cleanser composition stable to the human body.

It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Since the female cleaner manufactured by the method of manufacturing the female cleaner composition according to the present invention uses the natural material, it can produce the female cleaner through the finding of the natural material and can contribute to the industrial development.

10: drying step 20: extraction step
30: Filtration step 40: Sterilization step
50: Concentration step 60: Concentration correction step
70: mixing step 80: dilution step

Claims (8)

A method for producing an antiviral or an antimicrobial composition which selectively inhibits HSV1, HIV1, Neisseria gonorrhoeae, Mycoplasma hominis, Ureaplasma spp, Candida albicans or Escherichia coli among viral and infectious bacteria causing vaginitis,
10 to 40 parts by weight of an extract of Magnolia bark extract, 10 to 40 parts by weight of an extract of a centipede, 10 to 40 parts by weight of a centipede extract, 10 to 40 parts by weight of a licorice extract, 0.2 to 10 parts by weight of a dipotassium glycyrrhizinate (DPG) By weight of the antimicrobial composition to prepare an antiviral or antimicrobial composition The method according to claim 1,
A method for producing an antiviral or antimicrobial composition characterized by diluting a stock solution of a mixed composition and water at a weight ratio of 1: 5 to 1:20 in order to use the stock solution prepared by the above production method as a female cleanser A method for producing an antiviral or an antimicrobial composition which selectively inhibits HSV1, HIV1, Neisseria gonorrhoeae, Mycoplasma hominis, Ureaplasma spp, Candida albicans or Escherichia coli among viral and infectious bacteria causing vaginitis,
Drying step (10); Extraction step (20); Filtration step (30); A sterilization step 40; Concentration step 50; A density correction step (60); And mixing step (70); , Wherein:
A method for producing an antiviral or antimicrobial composition, which comprises mixing Rhizome extract, Magnolia bark extract, Centella asiatica extract, licorice extract, and DPG (Dipotassium Glycyrrhizinate) The method of claim 3,
Drying step 10 is a step of harvesting and drying natural materials such as yellowfin, magnolia bark, cinnabar and licorice;
Extracting step 20 is a step of boiling 10 kg of water in a drying step, boiling for 1 to 24 hours at 90 to 100 캜 for 1 to 24 hours at a temperature of 90 to 100 캜 until the total weight reaches about 7 Kg;
Filtration step (30) comprises filtering the extracted liquid from the extraction step with a cartridge filter (10 microns);
Sterilizing step 40 is a step of sterilizing the filtrate filtered at the filtration step to 95 캜;
The concentrating step 50 comprises concentrating the extract sterilized in the sterilization step to a concentration of 10-20 brix using a freeze dryer or a rotary evaporator;
The concentration correction step 60 includes diluting the extracts concentrated to be 10-20 brix in the concentration step with distilled water to dilute the extract so that the extract is 3 to 6 brix;
In the mixing step (70), the extracts adjusted to the same concentration (mg / L) in the concentration correction step are added to 10 to 60 parts by weight of the Rhizome extract, 10 to 40 parts by weight of the Magnolia bark extract, 10 to 40 parts by weight of the centipede extract, 10 to 40 parts by weight, and DPG (Dipotassium Glycyrrhizinate) in an amount of 0.2 to 10 parts by weight;
A method for producing an antiviral or antimicrobial composition delete 5. The method according to any one of claims 3 to 4,
A diluting step (80) of diluting the stock solution of the composition mixed in the mixing step (70) with the stock solution of the composition and water at a weight ratio of 1: 5 to 1:20 in order to use the composition prepared by the above- Or a method for producing an antiviral or antimicrobial composition The method according to any one of claims 1, 3, and 4,
The method for producing an antiviral or antimicrobial composition, wherein the antiviral or antimicrobial composition is selected from the group consisting of a solvent, a suspension, an emulsion, a tincture, a syrup, a spray, and an extract, delete

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