CN112245327A - Antibacterial composition, mouthwash and preparation method of mouthwash - Google Patents

Abstract

The application relates to the field of oral care, in particular to a bacteriostatic composition, mouthwash and a preparation method thereof. The bacteriostatic composition comprises glyceryl monocaprylate, cationic polymeric polypeptide and cyclodextrin; the mass ratio of the glyceryl monocaprylate to the cationic polymerized polypeptide to the cyclodextrin is (5-6): (8-10): (40-48). The application provides a bacteriostatic composition, not only improve the solubility and the dissolving speed of glyceryl monocaprylate in water, can also show improvement bactericidal effect, can also cover the pungent smell of glyceryl monocaprylate itself. The mouthwash prepared by the application adopts the compounding of the glycerol and the xanthan gum as the humectant, so that the moisturizing effect can be improved, the volatility of the menthol can be effectively reduced, and the mouthwash has no layering and no crystallization. The preparation method of the mouthwash provided by the application can promote cyclodextrin to better encapsulate glyceryl monocaprylate, and is good in stability.

Description

Antibacterial composition, mouthwash and preparation method of mouthwash

Technical Field

The application relates to the field of oral care, in particular to a bacteriostatic composition, mouthwash and a preparation method thereof.

Background

The oral cavity which is contacted with food every day is a hotbed of bacteria, and the bacteria in the oral cavity easily cause common oral diseases such as tooth decay and periodontal disease. More than six hundred kinds of bacteria exist in the oral cavity, and the common bacteria are phlegm-producing carbon dioxide cellulophilus, short lactic acid bacteria staphylococcus aureus, streptococcus mutans, oral streptococcus, escherichia coli, pseudomonas aeruginosa, streptococcus thermophilus and the like. The mouthwash can penetrate into various parts of the oral cavity, reduce bacteria and bacterial plaque causing halitosis, and improve the health of the gum.

At present, most of mouthwash products with bacteriostatic functions in the market are added with chemical bactericides such as triclosan and the like, but have strong side effects on human bodies; the synthesized chemical composition, namely chlorohexidine gluconate CHX or cetylpyridinium chloride CPC, needs to be added with alcohol, can cause irritation to oral mucosa of ordinary people, and can also cause tooth yellowing and influence appearance. Traditional Chinese medicine mouth wash is also widely used for inhibiting harmful bacteria in oral cavity, such as houttuynia cordata extract, dandelion extract, pomegranate rind extract and the like, but the effective components of the traditional Chinese medicine are volatile, oxidized and dark in color.

To maximize the efficacy of antiseptics in mouthwash, many attempts have been made by researchers in the field. For example, the glyceryl monocaprylate is safe, non-toxic, non-irritant, has spectral antibacterial property, and has complete inhibitory effect on bacteria, mold and yeast. However, glyceryl monocaprylate is insoluble in water and alcohol, greatly limiting the bactericidal effect of the mouthwash containing glyceryl monocaprylate.

In view of the above-mentioned related technologies, the applicant believes that there is a need to develop a bacteriostatic composition with high safety, high bacteriostatic effect and high stability.

Disclosure of Invention

In order to solve the problem of poor sterilization effect caused by the solubility of the glyceryl monocaprylate, the application provides a bacteriostatic composition, a mouthwash and a preparation method of the bacteriostatic composition.

In a first aspect, the application provides a bacteriostatic composition, which is realized by adopting the following technical scheme:

a bacteriostatic composition comprising glyceryl monocaprylate, a cationic polymeric polypeptide, and a cyclodextrin; the mass ratio of the glyceryl monocaprylate to the cationic polymerized polypeptide to the cyclodextrin is (5-6) to (8-10) to (40-48); the number of peptide bonds of the cationic polymerized polypeptide is 20-60.

By adopting the technical scheme, as the cationic polymerized polypeptide and the cyclodextrin are added, and the mass ratio of the glyceryl monocaprylate to the cationic polymerized polypeptide to the cyclodextrin is controlled to be (5-6) to (8-10) to (40-48), the solubility and the dispersibility of the glyceryl monocaprylate can be improved, and the sterilization effect can be obviously improved. The applicant believes that the cationic polymerized polypeptide is rich in cations, has good penetrating power on the biological membrane of oral pathogenic bacteria, and can be used as a carrier of glyceryl monocaprylate; the cyclodextrin has a structure with hydrophilic outer edge and hydrophobic inner cavity, and glyceryl monocaprylate can be embedded into the hydrophobic cavity of the cyclodextrin to form an inclusion complex, so that the solubility and the dissolution speed of the glyceryl monocaprylate in water are improved. In addition, the cationic polymerization polypeptide with the peptide bond number of 20-60 can inhibit the hydrolysis of the glyceryl monocaprylate into caprylic acid and glycerol and can partially cover the pungent smell of the glyceryl monocaprylate.

Preferably, the cationic polymeric polypeptide is polylysine; the polymerization degree of the polylysine is 25-30; more preferably, the polylysine is epsilon-polylysine.

By adopting the technical scheme, the polylysine containing free amino groups is adopted, belongs to the same polypeptide of a cationic polymer group, has a more regular molecular structure, is favorable for improving the penetrating power of cationic polymerization polypeptide on a biological membrane, and has the polymerization degree of 25-30, thereby being favorable for improving the synergistic effect and the carrier effect of the cationic polymerization polypeptide on the glyceryl monocaprylate. Especially epsilon-polylysine is easy to be absorbed by organisms, and the carboxyl and epsilon-amino form peptide bonds, the free amino is closer to the peptide bonds, the activity is stronger, and the more easily the epsilon-polylysine forms hydrogen bonds with other components such as cyclodextrin, and the like, so that the system stability is improved.

Preferably, the cyclodextrin is alpha-cyclodextrin and/or beta-cyclodextrin; more preferably, the cyclodextrin is β -cyclodextrin.

By adopting the technical scheme, the cyclodextrin embeds the glyceryl monocaprylate into the hydrophobic cavity of the cyclodextrin to form the inclusion complex. In the research process, the applicant finds that the molecular hole of the gamma-cyclodextrin is too large, and the polylysine is also partially wrapped and grafted, so that the sterilization effect is influenced; the alpha-cyclodextrin molecule is adopted, the hole is small, the molecular volume of the glyceryl monocaprylate is large, and part of the glyceryl monocaprylate is remained outside the cavity, so that the water solubility of the glyceryl monocaprylate is reduced; and the beta-cyclodextrin can be used for well wrapping the glyceryl monocaprylate and can cover the bad smell of the glyceryl monocaprylate.

Preferably, the beta-cyclodextrin is hydroxypropyl-beta-cyclodextrin and/or sulfobutyl-beta-cyclodextrin; more preferably, the cyclodextrin is hydroxypropyl- β -cyclodextrin.

By adopting the technical scheme, the water solubility of the glyceryl monocaprylate can be improved by adopting the beta-cyclodextrin to wrap the glyceryl monocaprylate, and when the mass ratio of the glyceryl monocaprylate to the beta-cyclodextrin is (5-6) to (40-48), the wrapped and connected material is easy to crystallize in water and cannot reach excellent solubility. The beta-cyclodextrin substituted by sulfobutyl or hydroxypropyl, especially the beta-cyclodextrin substituted by hydroxypropyl, is introduced, hydrogen bonds in beta-cyclodextrin molecules are opened by introducing the hydroxypropyl, an amorphous mixture is formed, the crystallinity of the cyclodextrin is reduced, meanwhile, the hydroxypropyl-beta-cyclodextrin can better inhibit the volatilization of menthol, and the hydrolysis resistance of epsilon-polylysine is improved.

In a second aspect, the present application provides a mouthwash, which adopts the following technical scheme:

a mouthwash comprises, by mass, 0.53-0.64% of the antibacterial composition, 0.05-0.06% of an aroma giving agent, 5-15% of a humectant and the balance of deionized water.

By adopting the technical scheme, the antibacterial composition is added into the mouthwash, so that the oral cavity is good in cleaning performance, the antibacterial composition has an excellent effect of inhibiting and killing pathogenic bacteria in the oral cavity, and the dental caries can be prevented; under the action of the aromatizing agent and the humectant, the smoothness and the mouthfeel of the mouthwash can be improved.

Preferably, the flavoring agent is selected from one or more of menthol, green tea essence, lemon essence, sweet orange essence and jasmine essence; more preferably, the flavorant is menthol.

By adopting the technical scheme, the menthol is adopted as the aromatizing agent, has the effects of clearing heat and purging fire, detoxifying and reducing swelling, and relieving sore throat and pain, can improve the fragrance of the mouthwash, but has low solubility in water and is volatile under the high-temperature condition.

Preferably, the humectant is a mixture of glycerin and xanthan gum; the mass ratio of the glycerol to the xanthan gum is 1: 0.00002-0.00004.

By adopting the technical scheme, the moisturizing effect can be improved by adopting the compounding of the glycerol and the xanthan gum as the moisturizing agent, the mass ratio of the glycerol to the xanthan gum is controlled to be 1: (0.00002-0.00004), the volatility of the menthol under the high-temperature condition can be effectively reduced, and the solubility of the menthol is improved.

In a third aspect, the application provides a preparation method of mouthwash, which adopts the following technical scheme:

a preparation method of mouthwash comprises the following steps:

s1: heating part of deionized water, dissolving glyceryl monocaprylate and cyclodextrin, and stirring in water bath; the mass ratio of the partial deionized water to the glyceryl monocaprylate is (80-120) to 1;

s2: and adding the cationic polymerization polypeptide, the aromatizing agent, the humectant and the rest deionized water, and mixing to obtain the antibacterial mouth wash.

Through adopting above-mentioned technical scheme, this application dissolves glyceryl monocaprylate and cyclodextrin in advance with the deionized water after the heating to the water bath stirring can promote the better packet of glyceryl monocaprylate of cyclodextrin, adds cationic polymerization polypeptide, perfuming agent, humectant again, can improve the stability of mouthwash.

Preferably, the heating temperature of the deionized water in the step S1 is 55-65 ℃.

By adopting the technical scheme, deionized water at 55-65 ℃ is adopted and water bath stirring is carried out at the temperature, the volatilization of menthol is not reduced while the solubility and the dissolving speed of the glyceryl monocaprylate are improved, and the synergistic effect of the cationic polymerization polypeptide and cyclodextrin on the glyceryl monocaprylate is ensured.

In summary, the present application has the following beneficial effects:

1. cationic polymerization polypeptide and cyclodextrin are added, the biomembrane of the oral pathogenic bacteria has good penetrating power, glyceryl monocaprylate can be embedded into a hydrophobic cavity of the cyclodextrin to form an inclusion complex, the solubility and the dissolving speed of the glyceryl monocaprylate in water are improved, the sterilization effect can be obviously improved, the hydrolysis of the glyceryl monocaprylate is inhibited, and the pungent smell of the glyceryl monocaprylate can be partially covered.

2. According to the application, polylysine containing free amino, especially epsilon-polylysine, is higher in activity, and is beneficial to improving the synergistic effect and the carrier effect of cationic polymerization polypeptide on glyceryl monocaprylate.

3. The beta-cyclodextrin is adopted, so that the glyceryl monocaprylate can be well coated, the bad smell of the glyceryl monocaprylate can be masked, and the antibacterial effect of the system is improved; especially hydroxymethyl substituted beta-cyclodextrin, reduces the crystallinity of the cyclodextrin, can better inhibit the volatilization of menthol and improve the hydrolysis resistance of epsilon-polylysine.

4. According to the application, glycerin and xanthan gum are compounded to serve as the humectant, so that the moisturizing effect can be improved, the volatility of menthol under a high-temperature condition can be effectively reduced, and the solubility of the menthol is improved.

5. According to the method, 55-65 ℃ deionized water is adopted to dissolve the glyceryl monocaprylate and the cyclodextrin in advance and stir in a water bath, the better encapsulation of the glyceryl monocaprylate by the cyclodextrin can be promoted, the volatilization of menthol can not be caused while the solubility and the dissolving speed of the glyceryl monocaprylate are improved, and the synergistic effect of the cationic polymerization polypeptide and the cyclodextrin on the glyceryl monocaprylate is ensured.

Detailed Description

In order to improve the sterilization effect of the mouthwash, the application adopts the safe, non-toxic and non-irritant glyceryl monocaprylate which plays a complete inhibition role on bacteria, mould and yeast. But the disadvantage is that glyceryl monocaprylate is insoluble in water, and the prepared mouthwash has poor stability, thereby affecting the sterilization effect of the mouthwash. In the research process, the applicant finds that the cationic polymerization polypeptide and the cyclodextrin are added, so that the solubility and the dispersibility of the glyceryl monocaprylate can be improved, and the sterilization effect can be obviously improved, but the crystallization phenomenon and the inclusion effect of the cyclodextrin can be influenced by different types and dosage of the cyclodextrin, and the sterilization effect and the stability of the mouthwash can be influenced. In addition, the addition of menthol may further mask the odor of glyceryl monocaprylate, but its menthol has low solubility in water and is volatile at high temperatures. The present application is based on this.

The present application will be described in further detail with reference to examples.

Examples

Examples 1 to 6

The following is an example of example 1, wherein the glyceryl monocaprylate is food grade and is purchased from David biosciences, Inc. of Jiangsu; the cationic polymerization polypeptide is YR-CP1, and the sequence of the YR-CP1 is as follows:

the cyclodextrin is gamma-cyclodextrin, and the CAS number is 17465-86-0; the menthol is L-menthol with CAS number of 2216-51-5; the xanthan gum is food grade and purchased from sinceri biotechnology limited of jenan.

The preparation method of the mouthwash provided by the embodiment 1 comprises the following steps: weighing glyceryl monocaprylate, cyclodextrin and deionized water (heated to 60 ℃) with the mass being 100 times that of the glyceryl monocaprylate according to the mass percent in the table 1, dissolving the glyceryl monocaprylate and the cyclodextrin, continuously stirring and mixing in a water bath at 60 ℃ for 30 minutes, adding the cationic polymerized polypeptide, menthol, glycerol, xanthan gum and the rest deionized water, and mixing to obtain the mouthwash.

As shown in Table 1, the mouthwashes of examples 1-6 differed only in the percentage by mass of the starting materials.

TABLE 1

Examples 7 to 8

Examples 7-8 differ from example 6 only in that:

example 7, except that: the cationic polymerization polypeptide is YR-CP2, and the sequence of the YR-CP2 is as follows:

example 8, except that: the cationic polymerization polypeptide is epsilon-polylysine, has the CAS number of 28211-04-3 and is purchased from Shanxi Chengming Biotech Co.

Examples 9 to 13

Examples 9-13 differ from example 8 only in that:

example 9, except that: the cyclodextrin is alpha-cyclodextrin, and the CAS number is 10016-20-3.

Example 10, except that: the cyclodextrin is beta-cyclodextrin, and the CAS number is 7585-39-9.

Example 11, except that: the cyclodextrin is carboxymethyl-beta-cyclodextrin, and the CAS number is 218269-34-2.

Example 12, except that: the cyclodextrin is sulfobutyl-beta-cyclodextrin, and the CAS number is 25167-62-8.

Example 13, except that: the cyclodextrin is hydroxypropyl-beta-cyclodextrin, and the CAS number is 128446-35-5.

Examples 14 to 17

Examples 14 to 17, like example 13, differ in that: the preparation method of the mouthwash is different.

Example 14, except that: weighing glyceryl monocaprylate, cyclodextrin and deionized water with the mass being 80 times of that of the glyceryl monocaprylate.

Example 15, except that: weighing glyceryl monocaprylate, cyclodextrin and deionized water with the mass being 120 times of that of glyceryl monocaprylate.

Example 16, except that: deionized water (heated to 55 ℃) having a mass 100 times that of glyceryl monocaprylate; the mixture was stirred continuously in a water bath at 55 ℃ for 30 minutes.

Example 17, except that: deionized water (heated to 65 ℃) with the mass 100 times that of the glyceryl monocaprylate; the mixture was stirred continuously in a water bath at 65 ℃ for 30 minutes.

Examples 18 to 20

Examples 18 to 20, which are the same as example 6, differ only in the kind of the starting material of the mouthwash.

Example 18, except that: the menthol is replaced by the jasmine essence which is food grade and is purchased from Shaanxi Yuanyou biological technology limited company.

Example 19, except that: the xanthan gum is replaced by sorbitol sugar, and the sorbitol sugar is food-grade and is purchased from Shijiazhuang Chunxin Biotechnology Co.

Example 20, except that: no xanthan gum.

Comparative example

Comparative examples 1 to 4

Comparative examples 1 to 4, which are identical to example 5, differ only in the starting material of the mouthwash.

Comparative example 1, except that: the YR-CP1 was replaced with γ -PGA with CAS number 25513-46-6, purchased from Wuhan Guanghua times Biotech, Inc.

Comparative example 2, except that: the mass percent of the cyclodextrin is 0.2%.

Comparative example 3, except that: non-cationic polymeric polypeptides.

Comparative example 4, except that: the mass percent of the glyceryl monocaprylate is 0.1%.

Comparative examples 5 to 7

Comparative examples 5 to 7, like example 5, differ only in the preparation of the mouthwash.

Comparative example 5, except that: deionized water (heated to 85 ℃) with a mass 100 times that of glyceryl monocaprylate; the mixture was stirred continuously in a water bath at 85 ℃ for 30 minutes.

Comparative example 6, except that: deionized water (heated to 35 ℃) having a mass 100 times that of glyceryl monocaprylate; the mixture was stirred continuously in a water bath at 35 ℃ for 30 minutes.

Comparative example 7, except that: directly mixing glyceryl monocaprylate, cyclodextrin, cationic polymerized polypeptide, an aromatizer, a humectant and deionized water at 60 ℃.

Performance test

The following performance tests were performed on the mouthwashes provided in examples 1 to 20 and comparative examples 1 to 7 of the present application;

the bacteriostatic effect is as follows: the mouthwashes provided in examples 1-20 and comparative examples 1-7 were each tested for Minimum Inhibitory Concentration (MIC),

the specific operation is as follows:

placing streptococcus mutans, leuconostoc mesenteroides and staphylococcus aureus on a broth culture medium for culturing for 24h, selecting a test colony with a typical form by using an inoculating loop, inoculating the test colony in 5mL of LB liquid culture medium, culturing at 37 ℃ for 200r/min to a logarithmic phase; taking a bacterial liquid, and adjusting the bacterial concentration to 10CFU/mL by using an LB liquid culture medium; the mouthwashes prepared in examples 1-20 and comparative examples 1-7 were diluted sequentially under aseptic conditions by the following method: sequentially adding the diluted solution into the 1 st to 11 th columns of a sterile 96-well plate from low to high, wherein each column contains 50 mu L of the diluted solution, the 12 th column contains a positive control, and adding the same amount of sterile water; then adding 50 mu L of bacterial liquid into each hole, sealing and uniformly mixing; at this time, the final mass concentrations of the mouth washes from the 1 st to the 11 th wells are 3200, 1600, 800, 400, 200, 100, 50, 25, 12.5, 6.25 and 3.125 μ g/mL (based on the mass of the paste) respectively; OD value at 0h was measured by a microplate reader, and then the 96-well plate was incubated at 37 ℃ for 24h and again measured. By comparing the OD values of the bacterial liquids at 0h and 24h, the minimum inhibitory concentrations of the mouthwashes of examples 1-20 and comparative examples 1-6 on streptococcus mutans, leuconostoc mesenteroides and staphylococcus aureus are determined; wherein, the minimum inhibitory concentration is less than 10ppm and is marked as A, the minimum inhibitory concentration is 10-20ppm and is not equal to 10ppm and is marked as B, the minimum inhibitory concentration is 20-40ppm and is not equal to 20ppm and is marked as C, the minimum inhibitory concentration is 40-70ppm and is not equal to 40ppm and is marked as D, the minimum inhibitory concentration is 70-100ppm and is not equal to 70ppm and is marked as E, the minimum inhibitory concentration is more than 100ppm and is marked as F, and the test result is shown in Table 2.

TABLE 2

Stability: the mouthwashes provided in examples 1-20 and comparative examples 1-7 were each allowed to stand at 25 ℃ for one month and observed for delamination and crystallization, and the results are shown in Table 3.

Odor: the mouth of the mouthwashes prepared immediately in examples 1 to 20 and comparative examples 1 to 7 were manually shot, respectively, and whether or not there was an offensive odor was smelled, and the test results are shown in Table 3.

Taste: the mouthwashes provided in examples 1-17, 19-20 and comparative examples 1-7 were rinsed after being left at 100 ℃ for 1 week, respectively, and were then smelled to determine whether they had a mint odor, as shown in Table 3.

TABLE 3

The present application is described in detail below with reference to the test data provided in tables 2 and 3.

As can be seen from examples 1-6 and comparative examples 2-4 of the application, the addition of the cationic polymerized polypeptide and the cyclodextrin improves the solubility of the system, controls the content of the cyclodextrin and the content of the glyceryl monocaprylate, and ensures that the prepared mouthwash has no layering and simultaneously improves the bacteriostatic effect on streptococcus mutans, leuconostoc mesenteroides and staphylococcus aureus.

From examples 6 to 8 of the present application, it is understood that the bactericidal effect of mouthwash can be improved by using epsilon-polylysine.

From examples 8 to 13 of the present application, it is known that compared with α -cyclodextrin, β -cyclodextrin can improve the antibacterial effect of mouthwash, and can also mask the unpleasant odor of glyceryl monocaprylate, but it is easy to delaminate and crystallize; the solubility of beta-cyclodextrin can be improved by adopting sulfobutyl-beta-cyclodextrin, the crystallinity of the cyclodextrin is reduced by adopting hydroxymethyl substituted beta-cyclodextrin, and the volatilization of menthol can be better inhibited.

From examples 13 and 18 to 20 of the present application, it is known that the glycerin and the xanthan gum are added to be compounded as the humectant, so that the volatility of the menthol under the high temperature condition can be effectively reduced, and the solubility of the menthol can be improved.

From the application example 5 and the comparative examples 5 to 7, the heated deionized water is used for dissolving the glyceryl monocaprylate and the cyclodextrin in advance and stirring in a water bath, so that the stability of the mouthwash can be improved; deionized water with the temperature of 55-65 ℃ is used for stirring in a water bath at the temperature, so that the dissolution and the dissolution speed of the glyceryl monocaprylate are improved, and the volatilization of the menthol is not reduced.

The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.

Claims (9)

1. A bacteriostatic composition, wherein the bacteriostatic composition comprises glyceryl monocaprylate, cationic polymerized polypeptide and cyclodextrin; the mass ratio of the glyceryl monocaprylate to the cationic polymerized polypeptide to the cyclodextrin is (5-6): (8-10): (40-48); the number of peptide bonds of the cationic polymerized polypeptide is 20-60.

2. The bacteriostatic composition according to claim 1, wherein the cationic polymeric polypeptide is polylysine; the polymerization degree of the polylysine is 25-30.

3. A bacteriostatic composition according to any one of claims 1-2 wherein said cyclodextrin is α -cyclodextrin and/or β -cyclodextrin.

4. A bacteriostatic composition according to claim 3 wherein said β -cyclodextrin is hydroxypropyl- β -cyclodextrin and/or sulfobutyl- β -cyclodextrin.

5. A mouthwash, which is characterized in that raw materials of the mouthwash comprise, by mass, 0.53-0.64% of the bacteriostatic composition according to any one of claims 1-4, 0.05-0.06% of an aroma giving agent, 5-15% of a humectant, and the balance being deionized water.

6. A mouthwash according to claim 5, wherein the humectant is a mixture of glycerol and xanthan gum; the mass ratio of the glycerol to the xanthan gum is 1: (0.00002-0.00004).

7. A mouthwash according to claim 5, wherein the flavour imparting agent is selected from one or more of menthol, green tea flavour, lemon flavour, orange flavour and jasmine flavour.

8. A process for the preparation of a mouthwash according to any of claims 5 to 7, comprising the following steps:

s1: heating part of deionized water, dissolving glyceryl monocaprylate and cyclodextrin, and stirring in water bath; the mass ratio of the partial deionized water to the glyceryl monocaprylate is (80-120): 1;

s2: and adding the cationic polymerization polypeptide, the aromatizing agent, the humectant and the rest deionized water, and mixing to obtain the antibacterial mouth wash.

9. The method as claimed in claim 8, wherein the deionized water of step S1 is heated at a temperature of 55-65 ℃.