CN111973551A - Mussel mucin antibacterial gel and preparation method thereof - Google Patents
Mussel mucin antibacterial gel and preparation method thereof Download PDFInfo
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- CN111973551A CN111973551A CN202010891870.8A CN202010891870A CN111973551A CN 111973551 A CN111973551 A CN 111973551A CN 202010891870 A CN202010891870 A CN 202010891870A CN 111973551 A CN111973551 A CN 111973551A
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- mussel mucin
- chitosan
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- sodium hyaluronate
- mussel
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
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- A61K31/13—Amines
- A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
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- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1767—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
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- A—HUMAN NECESSITIES
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Abstract
The invention discloses a mussel mucin antibacterial gel and a preparation method thereof, wherein the mussel mucin antibacterial gel comprises the following raw materials in percentage by mass: mussel mucin 1%, boric acid 0.2%, sodium hyaluronate 0.1% with 30 ten thousand molecular weight, chitosan 0.3-1.0%, menthol 0.1-0.5%, chlorhexidine acetate 0.2-0.5%, carbomer 9400.8-1.5%, ethanol 0.5-1.0%, ethylparaben 0.01-0.05%, triethanolamine 0.05-0.1%, and purified water 90-98%, wherein the total mass percentage of the components is 100%. The mussel mucin antibacterial gel is prepared by compounding mussel mucin, sodium hyaluronate, chitosan and the like, and has the effects of quickly repairing and moisturizing private mucous membranes, resisting oxidation, inhibiting bacteria and relieving itching, promoting wound healing, balancing flora and assisting in treating cervicitis, cervical erosion and various inflammations through the synergistic effect of the mussel mucin, the sodium hyaluronate and the chitosan.
Description
Technical Field
The invention belongs to the fields of medicine and personal care, and particularly relates to an antibacterial gel and a preparation method thereof.
Background
Data show that 86% of women worldwide do not know the severity of female personal care problems and neglect the importance of the personal care problems, and data show that the personal care problems can become invisible killers affecting female health in the future. Unfortunately, the personal care problem of 60% of women worldwide is further aggravated and becomes a disease, and many women will face double troubles from marital and health if the personal care problem cannot be solved and looked at in time.
Women like flowers, eager to have beautiful appearance and high-priced temperament are the beauty-pleasing nature of women. So that so many women have their efforts and time spent making up and shopping. The clothes can be more beautiful, make-up can be younger, but the most mysterious parts need to be maintained, and maintenance in private parts can make the whole person younger and healthier. If neglected, the personal health problem is not only hurt but also the family happiness is affected along with the time lapse, the age increase and the occurrence of the private health problem.
Disclosure of Invention
The invention aims to provide a mussel mucin antibacterial gel and a preparation method thereof, the mussel mucin antibacterial gel is prepared by compounding mussel mucin, sodium hyaluronate, chitosan and the like, and has the effects of quickly repairing and moisturizing private mucous membranes, resisting oxidation, inhibiting bacteria, relieving itching, promoting wound healing, balancing flora and assisting in treating cervicitis, cervical erosion and various inflammations through the synergistic effect of the mussel mucin, the sodium hyaluronate and the chitosan.
The technical scheme of the invention is as follows:
the mussel mucin antibacterial gel is characterized by being prepared from the following raw materials in percentage by mass: mussel mucin 1%, boric acid 0.2%, sodium hyaluronate 0.1% with 30 ten thousand molecular weight, chitosan 0.3-1.0%, menthol 0.1-0.5%, chlorhexidine acetate 0.2-0.5%, carbomer 9400.8-1.5%, ethanol 0.5-1.0%, ethylparaben 0.01-0.05%, triethanolamine 0.05-0.1%, and purified water 90-98%, wherein the total mass percentage of the components is 100%.
The mussel mucin antibacterial gel is characterized by being prepared from the following raw materials in percentage by mass: mussel mucin 1%, boric acid 0.2%, 30 ten thousand molecular weight sodium hyaluronate 0.1%, chitosan 0.4%, menthol 0.4%, chlorhexidine acetate 0.25%, carbomer 9401.0%, ethanol 0.8%, ethylparaben 0.03%, triethanolamine 0.08%, and purified water 95.74%, wherein the total mass percentage of the components is 100%.
A preparation method of mussel mucin antibacterial gel is characterized by comprising the following steps:
(1) cleaning and selecting required raw materials, and respectively weighing the raw materials according to the mass percentage for later use;
(2) dissolving the mussel mucin, the sodium hyaluronate and the chitosan weighed in the step (1) by 10 times of purified water respectively, and mixing to obtain transparent solution for later use;
(3) soaking carbomer and boric acid weighed in the step (1) in a proper amount of purified water for 24 hours, and uniformly stirring to obtain a soaking solution for later use;
(4) dissolving chlorhexidine acetate, ethylparaben and menthol with the ethanol weighed in the step (1) to obtain a dissolved solution for later use;
(5) adding the dissolved solution obtained in the step (4) into the soaking solution obtained in the step (3), uniformly stirring, adding the transparent solution obtained in the step (2) into the soaking solution obtained in the step (3), mixing and stirring, adding triethanolamine and stirring, and stirring for 4-5 hours in total to obtain a semi-finished bacteriostatic gel;
(6) passing through a colloid mill, inspecting the semi-finished product to be qualified, vacuumizing and filtering by using a vacuum pump, rotating at a rotating speed of-0.95 mpa for 10 minutes, and averaging for 10 minutes;
(7) filling 3 g/per bottle, wherein the net content allows negative deviation to be 9.0%;
(8) and packaging, inspecting finished products and warehousing.
The invention has the following positive effects:
(1) the mussel mucin antibacterial gel provided by the invention is simple in preparation method, convenient to use and suitable for large-scale popularization and use.
(2) In the field of female personal care, the mussel mucin antibacterial gel is a head-money-in-China cervical gel for word-eliminating repair made of mussel mucin by adopting the material characteristics.
(3) The mussel mucin antibacterial gel provided by the invention is scientific in formula, and the used raw materials and auxiliary materials are safe and non-irritating, and have the effects of quickly repairing and moisturizing private mucous membranes, resisting oxidation, inhibiting bacteria, relieving itching, promoting wound healing, balancing flora and assisting in treating cervicitis, cervical erosion and various inflammations.
Drawings
Fig. 1 is a flow chart of a preparation process of the mussel mucin bacteriostatic gel.
Fig. 2 is a comparison chart of the experiment for repairing mucosal injury, wherein a is a mucosal image of mussel mucin, and b is a mucosal image after mussel mucin treatment.
Table 1 is a data table of raw materials in the mussel mucin bacteriostatic gel of the invention
Performance test of mussel mucin bacteriostatic gel
1. And (3) testing the irritation of the mussel mucin antibacterial gel to the vaginal mucosa.
2. And (3) testing a sample: the mussel mucin antibacterial gel is gel, and an original sample is taken as a tested object.
3. Experimental animals: female New Zealand white rabbits 6 were provided by Dialep, Seisan, Bio-medicine, Inc., weighing 2.1-2.4 kg.
Common level, production license number: SCXK (shan) > 2019002. The feed source is the same as above.
4. Animal room environment: ordinary ambient temperature: 18 ℃ -29 ℃, relative humidity: 40-70 percent of the total weight of the animal experiment, and the license number used by the experimental animal is SYXK (shan) 2019-.
5. The test method comprises the following steps: the 6 non-estrus domestic rabbits were randomly divided into 2 groups, i.e., a contaminated group and a control group, each group consisting of 3 rabbits, and during the test, the animals were fixed on their backs, the perineum and vaginal opening were exposed, the catheter was gently inserted into the vagina of 4cm-5cm, 2g of the test substance was slowly injected with a syringe, and the animals in the control group were treated with physiological saline in the same manner. Killing animals 24h after the last infection, taking out the complete vagina by adopting an air embolism method, carrying out laparotomy, carrying out visual observation on whether congestion, edema and other manifestations exist, then putting the vagina into 10% formalin solution for fixation for more than 24h, selecting tissues at two ends and the center of the vagina for flaking, carrying out histopathological examination after HE staining, and carrying out evaluation according to the vagina mucosa irritation reaction scoring standard.
6. The test results are shown in Table 2
TABLE 2 results of the experiment on the irritation of mussel mucin gel to rabbit's vaginal mucosa
7. Evaluation of
According to the vaginal mucosa stimulation intensity powder standard in the technical Specification for disinfection products, the vaginal mucosa stimulation index of the mussel mucin bacteriostatic gel is 0.22, and the stimulation reaction intensity is nonirritant.
Table 3 is a statistical table of the results of the female cervicitis gynecological disease irritation test
As can be seen from the table 3, no matter the mussel mucin antibacterial gel is applied to the vaginal mucosa without cervicitis and gynecological diseases or the vaginal mucosa with cervicitis and gynecological diseases, no stinging sensation is caused to a person in the using process, and the skin is free from swelling and irritation after 24 hours. The mussel mucin antibacterial gel provided by the invention is basically non-irritant to vaginal mucosa.
Secondly, bacteriostasis test of the mussel mucin bacteriostasis gel
1. Test equipment
1.1 test strains: escherichia coli (8099). Cultured for 7 generations. Staphylococcus aureus (ATCC6538) was cultured for 7 generations, Candida albicans (ATCC10231) was cultured for 7 generations. Provided by China center for preservation and management of industrial microbial species.
1.2 sample: mussel mucin bacteriostatic gel.
1.3 vector: the filter paper sheet was 10cm by 1.0 cm.
1.4 nutrient agar, Sha's agar was provided by Beijing Luqiao technology GmbH.
1.50.03 mol/L Phosphate Buffer Solution (PBS).
1.6 sterile graduated pipette, plate, test tube, thermostatic wall culture box (type 303-3), etc.
2. Inspection method
2.1 according to GB15979-2002 hygienic Standard for Disposable sanitary articles).
2.2 the bacteriostatic test is a carrier test, the test temperature is 20 +/-1 ℃, and the test is repeated three times.
3. Test results
The mussel mucin antibacterial gel has the effect of 5mim, the average antibacterial rate on escherichia coli (8099) reaches 100%, the average antibacterial rate on staphylococcus aureus (ATCC6538) reaches 100%, and the average antibacterial rate on candida albicans (ATCC10231) reaches 100%. The results are shown in tables 4, 5 and 6.
TABLE 4 bacteriostatic effect of mussel mucin gel on E.coli
Note: the average number of producing colonies in the positive control group was 2.66X 104cfu/disc, negative control sterile growth.
TABLE 5 bacteriostatic Effect of mussel mucin gel on Staphylococcus aureus
Note: the average number of producing colonies in the positive control group was 2.27X 104cfu/disc, negative control sterile growth.
TABLE 6 bacteriostatic Effect of mussel mucin gel on Candida albicans
Note: the average number of producing colonies in the positive control group was 2.37X 104cfu/disc, negative control sterile growth.
Toxicity test of mussel mucin bacteriostatic gel
1. Preparation before testing
60 mice are treated by electric hair clippers before administration, the hair removal area of the mice along the two sides of the spine is 2 multiplied by 2cm, the hair removal area of the mice is observed to be damaged due to hair removal after 24 hours, and the mice cannot be used as a complete skin group experiment if the skin is damaged; the 60 mice were divided into 6 groups of 10 mice each, each group having half male and female, groups 1-3 being intact skin groups, groups 4-6 being damaged skin groups. Among them, 30 mice in the group with damaged skin were subjected to skin scratch with fine sandpaper in the depilated area, with no damage to the dermis and slight bleeding as the standard. Group 1 was an intact skin blank control group; group 2 is a group of intact skin low dose medications; group 3 is a group of intact skin high dose medications. Group 4 is a damaged skin blank control group; group 5 is a group of low dose drugs for damaged skin; group 6 is a group of high dose drugs for damaged skin.
2. Test procedure
Coating 0.5g of the mussel mucin antibacterial gel consisting of the raw materials described in example 1 on a low-dose medicine application group in a depilation area, coating 1g of the mussel mucin antibacterial gel consisting of the raw materials described in example 1 on a high-dose medicine application group in the depilation area, cleaning an administration part by using a sodium chloride solution with the concentration of 0.9% after 24 hours of coating, then continuing to feed according to a conventional feeding method, simultaneously recording the time of toxicity appearance and disappearance, removing 1h of the medicine as a first observation point, then respectively carrying out observation and recording for 8h, 24h and 48h, and if the toxicity appears after 48h, continuing to observe and record. Attention is paid to the red and swollen degree of the drug-coated area of the white mouse, the hair color, the body weight, the change of eyes and mucous membranes, the activity of limbs and other performance manifestations.
3. Test results
Through observation, the skin luster of the medicine-coated areas of the mice of the groups 2, 3, 5 and 6 is basically not different from that of the groups 1 and 4 of the control group, the mice can move freely, the eyes and the mucous membranes are not abnormal, the weight change is kept in a reasonable range, and no mice die, which shows that the mussel mucin bacteriostatic gel has no acute toxic reaction.
4. Wound healing test
4.1, preparation before testing
The method comprises the steps of removing hair areas of 16 mice with SPF (specific pathogen free) grades, the body weights of 150 +/-10 g and half of males and females along two sides of a spine by using an electric hair clipper before administration, then carrying out anesthesia on the mice, wherein the dosage of an anesthetic is the same standard, marking 1 circle with the diameter of 1cm at the hair removal position, disinfecting the circle with iodine, and then removing the whole skin of the marked two circles by using a medical clipper to form two circular wound surfaces.
4.2 test procedure
The mice are divided into 2 groups, each group comprises 8 mice, each mouse is half male and female, the 1 st group is the mussel mucin bacteriostatic gel prepared from the raw materials described in the example 1, and the 2 nd group is common external normal saline. The two groups of mice begin to take the medicine after 0.5 hour after the wound, the interval time is 4 hours, the medicine is continuously taken for 24 days, and the dosage is 1 mL; the mental state of the rats and the time to wound onset scab, as well as the time to wound onset scab, were recorded daily.
Table 7 shows the healing time of the wounds of different groups of mice
| Serial number | Scab formation time (h) | Decrustation time (h) |
| Group 1 | 75 | 340 |
| Group 2 | 165 | 503 |
4.3 test results
As can be seen from Table 7, the eschar time and the eschar time appeared to be positively correlated, and the eschar time was earlier in group 1 with the earlier eschar time, and the eschar skin at the wound was smoother. Compared with normal saline, the mussel mucin antibacterial gel prepared from the raw materials in the embodiment 1 of the invention can effectively shorten the scabbing time and the decrustation time, and the mussel mucin antibacterial gel prepared from the raw materials in the embodiment 1 of the invention has better effects of diminishing inflammation and promoting wound healing.
5. Moisture retention test
The instrument is a silica gel dryer, the ambient temperature is 25 ℃, and the experimental object is the mussel mucin bacteriostatic gel consisting of the raw materials in the embodiment 1.
The test process comprises the following steps: weigh the vial by a weight denoted CAir conditioner2g of the mussel mucin bacteriostatic gel prepared from the raw materials described in example 1 was placed in a weighing bottle, and the mass of the gel was accurately weighed and recorded as C0Placing in a silica gel dryer, standing for 1, 3, 5, 7, 9, 11, 13, 15, 17, 21, and 24 hours (h), weighing again, and recording as Cn；
The water loss rate is calculated as follows:
water loss (%) - (C)0-Cn)/(C0-CAir conditioner) X 100%, in the formula,Cair conditionerEmpty weighing bottle mass, C0The mass of a weighing bottle for holding 2g of the mussel mucin bacteriostatic gel consisting of the raw materials described in example 1, CnThe mass of the weighing bottle for containing the mussel mucin antibacterial gel after being placed for n hours.
Moisture retention (%) - (100% water loss (%)
The lower the water loss rate, the better the moisturizing effect.
Table 8 is a moisture retention test data table
As can be seen from table 8, the moisture retention of the mussel mucin antibacterial gel of the invention can still reach 98.80% after 24 hours, and the moisture retention rate of the commonly used humectant glycerin in skin care products is as follows: moisture retention of 96% and L-lysine hydrochloride: 87.6% (data from the literature, "moisture absorption and retention and kinetics of laminine"). The mussel mucin bacteriostatic gel has a good moisturizing effect.
6. Control of mucosal injury repair experiment
Mussel mucin contains a large amount of lysine and carries a high amount of positive charge at physiological ph values. The human tissues are negatively charged, the proteins are quickly adsorbed to the surfaces of the cells by positive and negative attraction to form a non-continuous micro-nano biological scaffold, and the support and the direction are provided for the adherence and crawling of the cells through the electrostatic physical action, so that the tissue reconstruction is facilitated, the wound healing is promoted, and the bacteriostatic action is realized. Has good effect on mild and moderate cervical erosion.
7. Anti-inflammatory
The characteristic has obvious effect on aseptic vaginitis and senile vaginitis.
8. Oxidation resistance
With age, oxygen radicals gradually destroy body cellular tissues, leading to aging. Mussel mucin contains 10% of dopa group, and is the only known protein with high content of dopa group, so that it can resist aging, prevent the generation of new melanin, make mucosa tender, and quickly relieve pruritus.
Fourth, applicable therapeutic scenario
1. Postoperative maintenance
And (3) postoperative care: within 7 days after treatment, one part of the Chinese medicinal composition is used every day to promote wound healing and repair and maintain the health of the wound.
2. Domestic maintenance
Indications are as follows: wrinkle disappearance, mucosa thinning, and other symptoms of intimate aging, including dryness, pruritus, odor, and laxity. Promoting the youthfulness of privacy, inhibiting melanin, and increasing sensitivity, thereby improving the pleasure of sexual life. (recommended daily maintenance 2-3 per week)
3. Sexual life
The use before sexual life can form a barrier to prevent virus infection and has lubricating effect to relieve pain of sexual life; can repair mucosa injury after sexual life, recover vaginal environment, and has effects of relieving and caring.
4. Gynecological application
Indications are as follows: repairing mucosa wound, aseptic/senile vaginitis, cervical erosion, and repairing and healing induced abortion and puerperal abortion.
5. Daily care
The people paying attention to daily maintenance can be used for daily private nursing.
The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and is not intended to limit the invention to the particular forms disclosed. For those skilled in the art to which the invention pertains, several simple deductions or substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.
Claims (4)
1. The mussel mucin antibacterial gel is characterized by being prepared from the following raw materials in percentage by mass: mussel mucin 1%, boric acid 0.2%, sodium hyaluronate 0.1% with 30 ten thousand molecular weight, chitosan 0.3-1.0%, menthol 0.1-0.5%, chlorhexidine acetate 0.2-0.5%, carbomer 9400.8-1.5%, ethanol 0.5-1.0%, ethylparaben 0.01-0.05%, triethanolamine 0.05-0.1%, and purified water 90-98%, wherein the total mass percentage of the components is 100%.
2. The mussel mucin bacteriostatic gel according to claim 1, which is prepared from the following raw materials in percentage by mass: mussel mucin 1%, boric acid 0.2%, 30 ten thousand molecular weight sodium hyaluronate 0.1%, chitosan 0.4%, menthol 0.4%, chlorhexidine acetate 0.25%, carbomer 9401.0%, ethanol 0.8%, ethylparaben 0.03%, triethanolamine 0.08%, and purified water 95.74%, wherein the total mass percentage of the components is 100%.
3. A method for preparing a mussel mucin bacteriostatic gel according to claim 1 or 2, which comprises the following steps:
(1) cleaning and selecting required raw materials, and respectively weighing the raw materials according to the mass percentage for later use;
(2) dissolving the mussel mucin, the sodium hyaluronate and the chitosan weighed in the step (1) by 10 times of purified water respectively, and mixing to obtain transparent solution for later use;
(3) soaking carbomer and boric acid weighed in the step (1) in a proper amount of purified water for 24 hours, and uniformly stirring to obtain a soaking solution for later use;
(4) dissolving chlorhexidine acetate, ethylparaben and menthol with the ethanol weighed in the step (1) to obtain a dissolved solution for later use;
(5) adding the dissolved solution obtained in the step (4) into the soaking solution obtained in the step (3), uniformly stirring, adding the transparent solution obtained in the step (2) into the soaking solution obtained in the step (3), mixing and stirring, adding triethanolamine and stirring, and stirring for 4-5 hours in total to obtain a semi-finished bacteriostatic gel;
(6) passing through a colloid mill, inspecting the semi-finished product to be qualified, and performing vacuum pumping and filtration by using a vacuum pump;
(7) filling 3 g/per bottle, wherein the net content allows negative deviation to be 9.0%;
(8) and packaging, inspecting finished products and warehousing.
4. The method for preparing a mussel mucin bacteriostatic gel according to claim 3, wherein the method comprises the following steps: and (6) the vacuum degree of the step (6) is 0.95mpa, the rotating speed is 45 revolutions for 10 minutes, and the average value is 10 minutes.
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