Mouth wash composition capable of enhancing antibacterial efficiency

Abstract

The invention provides a mouthwash composition which comprises the following effective components: lactobacillus salivarius, domiphen, a pH regulator, a stabilizer and a preservative, the content of the lactobacillus salivarius is 107-1011 CFU / mL, the concentration of the domiphen is 0.05-0.2 wt%, the pH value of the mouthwash composition is regulated to 6.8-7.2 by the pH regulator, and the balance of water, the mouthwash composition can achieve an improved antibacterial effect in an oral environment, and can achieve a lasting antibacterial effect under the condition of maintaining the balance of oral microbial communities.

Description

Technical Field

[0001] The invention relates to the field of oral care, and in particular to a mouthwash composition with enhanced antibacterial efficacy. Background Art

[0002] As an important part of oral care, mouthwash has evolved from simply refreshing breath to cleaning the mouth, assisting in removing food residues, reducing oral bacteria, and preventing oral diseases. The active ingredients it contains can not only quickly improve the oral environment, but also penetrate into the gaps between teeth and hard-to-reach areas of the mouth to help maintain the health of teeth and gums. With the accelerated pace of modern life, people are paying more and more attention to oral health and personal image, and the use of mouthwash has become an indispensable part of daily life.

[0003] The antibacterial effect of mouthwash is one of its most critical functions. Effective antibacterial ingredients can not only quickly reduce the number of bacteria in the mouth and reduce the risk of infection caused by pathogenic microorganisms, but also inhibit bacterial metabolic activities and reduce the production of bad odors such as sulfides, thereby creating a clean and healthy microenvironment in the mouth. Among the many antibacterial agents, domiphene has become a preferred ingredient in mouthwash products due to its excellent antibacterial properties and safety. As a cationic surfactant, domiphene can quickly bind to the bacterial cell wall and destroy its structure, thereby achieving the purpose of quickly removing oral pathogens. In addition, the broad-spectrum antibacterial properties of domiphene enable it to effectively fight a variety of oral pathogens, including Streptococcus mutans that causes caries and Porphyromonas gingivalis that causes periodontal disease.

[0004] Although the antibacterial effect of domiphene in mouthwash is significant, a major problem it faces is the lack of durability of the antibacterial effect. This deficiency is not limited by the properties of domiphene itself, but is closely related to the complexity of the oral environment. The oral cavity is an ecosystem full of microorganisms, and its pH value, temperature, saliva composition and flow may affect the efficacy of antibacterial agents. In particular, once the structure of the oral flora changes and a slightly acidic environment with a pH below 6.5 appears, the activity of domiphene will be inhibited and it will not be able to maintain an effective antibacterial level for a long time.

[0005] In addition, the flow and flushing of saliva in the mouth, as well as the pH changes caused by daily diet, may quickly dilute or neutralize the antibacterial components in mouthwash, further reducing its durability. Long-term reliance on mouthwash with a single antibacterial mechanism may also lead to changes in the structure of the oral microbial community and affect the natural defense capabilities of the oral cavity.

[0006] Therefore, developing a mouthwash that can adapt to the complex environment of the oral cavity and achieve a lasting antibacterial effect is a key requirement for current technological development. This mouthwash needs to be able to overcome the challenges of the oral environment, such as pH fluctuations, saliva flow and other factors, while providing stable and lasting antibacterial protection without interfering with the balance of the natural oral flora. This requires not only the improvement of existing antibacterial agents, but also the exploration of new formulas and mechanisms to achieve comprehensive maintenance of oral health. Summary of the invention

[0007] The present invention aims to provide a mouthwash that can adapt to the complex environment of the oral cavity and achieve a long-lasting antibacterial effect. The mouthwash provided by the present invention can achieve an improved antibacterial effect in the oral environment and achieve a long-lasting antibacterial effect while maintaining the balance of the oral microbial community. In addition, the present invention also provides a preparation method of the mouthwash and its application in oral care. The details are as follows:

[0008] The present invention provides a mouthwash composition, comprising the following effective ingredients: Lactobacillus salivarius, domiphene, a pH regulator, a stabilizer and a preservative, wherein the content of Lactobacillus salivarius is 10 based on the total weight of the mouthwash composition. 7 Up to 10 11 CFU / mL, the concentration of domiphene is 0.05wt% to 0.2wt%, the pH adjuster adjusts the pH value of the mouthwash composition to 6.8 to 7.2, and the balance is water.

[0009] In one embodiment, the pH adjuster is selected from one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium bicarbonate and sodium carbonate.

[0010] In one embodiment, the concentration of the stabilizer is 0.05 wt % to 0.1 wt %, and the stabilizer is selected from one or more of sodium carboxymethyl cellulose, trehalose, xanthan gum, and polyvinyl pyrrolidone.

[0011] In one embodiment, the concentration of the preservative is 0.05 wt % to 0.1 wt %, and the preservative is selected from one or more of methylparaben, propylparaben, phenoxyethanol, and nitrobenzoate.

[0012] In one embodiment, the mouthwash composition further comprises fluoride.

[0013] In one embodiment, the mouthwash composition further comprises vitamin C and / or vitamin E as antioxidants.

[0014] In one embodiment, the Lactobacillus salivarius is in the form of microcapsules; the specific method is as follows:

[0015] a. suspending Lactobacillus salivarius in an aqueous solution containing a stabilizer to form an initial suspension;

[0016] b. adding an aqueous solution of microcapsule wall material to the initial suspension, and mixing to form a mixed suspension having a stabilizer and a wall material;

[0017] c. preparing microcapsules by emulsification or spray drying;

[0018] The stabilizer is selected from one or more of trehalose, xanthan gum, sodium carboxymethyl cellulose or polyvinyl pyrrolidone;

[0019] The microcapsule wall material is selected from one or more of gelatin, modified starch, cellulose derivatives or mixtures thereof.

[0020] In one embodiment, the particle size of the Lactobacillus salivarius microcapsule is 5-100 μm, preferably 10-80 μm, and more preferably 20-60 μm.

[0021] The present invention also provides a method for preparing the mouthwash composition, comprising the following steps:

[0022] 1) Activating the strain to form a Lactobacillus salivarius culture solution;

[0023] 2) mixing the pH adjuster, stabilizer and preservative with the domiphene solution,

[0024] 3) under aseptic conditions, adding the Lactobacillus salivarius culture solution in step 1) to the domiphene mixed solution in step 2), and dispersing the mixture evenly at a low temperature of 0-10° C. to form the mouthwash composition, so that the content of Lactobacillus salivarius in the final composition is 10 7 Up to 10 11 CFU / mL, and the concentration of domiphene was 0.05wt% to 0.2wt%.

[0025] Furthermore, the method for preparing the mouthwash composition also includes a method for microencapsulating Lactobacillus salivarius, specifically, after step 1), further including the following steps:

[0026] a. suspending Lactobacillus salivarius in an aqueous solution containing a stabilizer to form an initial suspension;

[0027] b. adding an aqueous solution of microcapsule wall material to the initial suspension, and mixing to form a mixed suspension having a stabilizer and a wall material;

[0028] c. Prepare microcapsules by emulsification or spray drying,

[0029] And after Lactobacillus salivarius is microencapsulated, no stabilizer is added in step 2).

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] First, the pH value of the mouthwash composition is adjusted to the range of 6.8-7.2 by adding a neutral pH regulator into the mouthwash. Domiphene can more effectively penetrate the bacterial cell wall in a neutral environment and exert its broad-spectrum antibacterial properties, thereby maximizing its antibacterial effect.

[0032] Secondly, the present invention adds Lactobacillus salivarius to the mouthwash. As a probiotic, Lactobacillus salivarius can quickly respond to and continuously improve the slightly acidic environment of the oral cavity, and cooperate with a pH regulator to create an environment for the oral cavity that is not conducive to the growth of pathogenic bacteria, so that domiphene can continue to demonstrate its broad-spectrum antibacterial properties for a long time.

[0033] In summary, the present invention maximizes the antibacterial effect and long-lasting efficacy of domiphene through the synergistic effect of Lactobacillus salivarius, domiphene and the pH regulator, so that the mouthwash not only provides protection in the short term, but also helps to improve the long-term inhibition of harmful bacteria (such as Streptococcus mutans, Porphyromonas gingivalis, etc.) in the oral cavity, thereby achieving long-term maintenance of oral health. DETAILED DESCRIPTION

[0034] Although the present invention has been described to a certain extent, it is obvious that appropriate changes in various conditions can be made without departing from the spirit and scope of the present invention. It is understood that the present invention is not limited to the embodiments described, but belongs to the scope of the claims, which includes equivalent replacements of each factor described.

[0035] The present invention provides a mouthwash composition, comprising the following effective ingredients: Lactobacillus salivarius, domiphene, a pH regulator, a stabilizer and a preservative, wherein the content of Lactobacillus salivarius is 10 7 Up to 10 11 CFU / mL, the concentration of domiphene is 0.05wt% to 0.2wt%, the pH adjuster adjusts the pH value of the mouthwash composition to 6.8 to 7.2, and the balance is water.

[0036] In the present invention, a pH regulator is added to the original antibacterial domiphene mouthwash composition to make the mouthwash neutral, so that the antibacterial activity of domiphene is maintained at a high level, and the growth of harmful bacteria (such as cariogenic bacteria) can also be reduced. These bacteria usually reproduce more actively in an acidic environment. The pH regulator is selected from one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium bicarbonate and sodium carbonate. These pH regulators are neutral pH buffer reagents that can stabilize the pH value of the mouthwash within the range of 6.8-7.2, and are preferably a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate.

[0037] Lactobacillus salivarius is also added to the composition. Lactobacillus salivarius is a Gram-positive bacillus with probiotic activity. Natural Lactobacillus salivarius strains are widely present in the human oropharyngeal gastrointestinal tract (OGT), and have the effects of inhibiting pathogenic bacteria and stimulating immune cells to secrete anti-allergic related cytokines. The researchers found that although Lactobacillus salivarius grows under facultative anaerobic or absolute anaerobic conditions, the optimal growth temperature is 30-43°C, and the optimal pH value is 5.0-5.5 or lower, it also has good pH adaptability and can tolerate a certain range of pH changes, allowing it to remain stable in oral care products and provide a probiotic effect in a neutral (pH value in the range of 6.8-7.2) oral environment. The experimental results show that when an appropriate amount of Lactobacillus salivarius is added in a neutral environment, compared with other flora such as Lactobacillus casei, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus acidophilus, etc., Domifen can continue to reflect its broad-spectrum antibacterial properties for a long time. The reason for this may be that the metabolites produced by Lactobacillus salivarius under neutral conditions have little effect on the pH value of the oral cavity, which can help maintain the neutral state of the oral environment for a long time and help stimulate the antibacterial effect of domiphene. The antibacterial effect of domiphene is not limited to the elimination of pathogenic microorganisms. The environment created after the removal of pathogenic microorganisms is also conducive to the growth and colonization of Lactobacillus salivarius. Domiphene reduces or eliminates microorganisms that compete with probiotics, providing more favorable growth conditions for Lactobacillus salivarius. The colonization of Lactobacillus salivarius on the oral mucosa helps it to continue to play a role, prolongs the antibacterial effect of domiphene, and helps to continuously inhibit the regeneration of pathogenic microorganisms, thereby further prolonging the protective effect of mouthwash.

[0038] In the mouthwash composition, the content of Lactobacillus salivarius is 10 7 Up to 10 11 CFU / mL, preferably 10 8 Up to 5×10 10 CFU / mL, more preferably 5×10 8 Up to 10 10 CFU / mL. Within this range, it can ensure that there is a sufficient number of live Lactobacillus salivarius to effectively improve the slightly acidic environment of the oral cavity and provide a neutral environment. If the content is too low, the number of live bacteria is insufficient and it cannot improve the oral environment. If the content is too high, the metabolic activity of Lactobacillus salivarius is too vigorous, thereby affecting the balance of the natural microbial community in the oral cavity and also affecting the taste of the mouthwash.

[0039] In one embodiment, the concentration of domiphene is 0.05wt% to 0.2wt%, preferably 0.08-0.15wt%. Too high a concentration may cause irritation to the oral mucosa, causing discomfort, and will not improve the bactericidal effect, while too low a concentration may hardly achieve an effective antibacterial effect.

[0040] In one embodiment, the concentration of the stabilizer is 0.05wt% to 0.1wt%, and is selected from one or more of sodium carboxymethylcellulose, trehalose, xanthan gum, and polyvinyl pyrrolidone. These stabilizers help maintain the stability and uniformity of the mouthwash and prevent the effective ingredients from decomposing or precipitating. The stabilizer can also be used to embed Lactobacillus salivarius to further prevent the effective ingredients from decomposing or metabolizing before use, and is preferably sodium carboxymethylcellulose.

[0041] In one of the schemes, the concentration of the preservative is 0.05wt% to 0.1wt%, selected from one or more of methyl paraben, propyl paraben, phenoxyethanol, and nitrobenzoate. Since mouthwash usually contains water and other nutrients, these ingredients easily provide a hotbed for the growth of microorganisms, causing the product to deteriorate or produce harmful microorganisms. Adding preservatives can ensure the long-term stability and safety of the mouthwash composition. In the formula of mouthwash, the concentration range of the above preservatives (0.05wt% to 0.1wt%) is the optimal concentration range determined by experiments, which can not only ensure the microbial stability of the product during the shelf life, but also will not cause unnecessary stimulation or allergic reactions to the oral mucosa of consumers. In addition, this concentration range also meets the restrictions on the use of preservatives in cosmetics and oral care products in most countries and regions. When selecting a preservative, it is also necessary to consider its compatibility with other ingredients in the product, as well as the impact on the pH value and taste of the product, to ensure that the product can provide a good user experience while maintaining stability.

[0042] In one of the schemes, the mouthwash composition also contains fluoride. Fluoride is added to enhance the resistance of teeth to acid and prevent the occurrence of caries. Sodium fluoride and ammonium fluoride are commonly used fluoride additives, which can react with the hydroxyapatite of teeth to form more acid-resistant fluoroapatite, thereby improving the anti-caries ability of teeth. Sodium fluoride is preferred because it has good solubility and stability and is easy to mix with other ingredients. Based on the mouthwash composition, the content is 0-0.05wt%, preferably 0.01wt%-0.03wt%.

[0043] In one embodiment, the mouthwash composition further comprises vitamin C and / or vitamin E as antioxidants. The addition of vitamin C (ascorbic acid) and vitamin E (tocopherol) helps neutralize free radicals and protect oral tissues from oxidative damage. Vitamin C also helps promote gum health and reduce inflammatory responses; vitamin E can protect cell membranes from damage by oxidants. Based on the mouthwash composition, the content of vitamin C and / or vitamin E is 0-0.01wt%, preferably 0.001wt%-0.005wt%.

[0044] In one embodiment, the Lactobacillus salivarius is in the form of microcapsules; further, the Lactobacillus salivarius is embedded in the microcapsules, and the specific method is as follows:

[0045] a. suspending Lactobacillus salivarius in an aqueous solution containing a stabilizer to form an initial suspension;

[0046] b. adding an aqueous solution of microcapsule wall material to the initial suspension, and mixing to form a mixed suspension having a stabilizer and a wall material;

[0047] c. preparing microcapsules by emulsification or spray drying;

[0048] The stabilizer is selected from one or more of trehalose, xanthan gum, sodium carboxymethyl cellulose or polyvinyl pyrrolidone;

[0049] The microcapsule wall material is selected from one or more of gelatin, modified starch, cellulose derivatives or mixtures thereof, and the amount is preferably enough to encapsulate Lactobacillus salivarius. In the final composition, the content is 0.5wt%-2wt%.

[0050] According to subsequent experiments, microencapsulation of Lactobacillus salivarius helps to further improve the antibacterial and long-term antibacterial properties of the mouthwash composition. It can be inferred that the advantages of microencapsulation of Lactobacillus salivarius include improving the stability of probiotics during production, storage and transportation, protecting probiotics from the influence of external environment such as humidity, temperature change and oxygen, thereby maintaining its activity and quantity. And microencapsulation also helps to control the release rate of Lactobacillus salivarius in the oral cavity, ensures its colonization effect on the oral mucosa, and prolongs the action time of probiotics. In addition, the selection of microcapsule wall material and stabilizer can further improve the survival rate of Lactobacillus salivarius, enhance its tolerance to the acidic environment of the oral cavity, and then improve the overall efficacy and safety of mouthwash. By microencapsulation of Lactobacillus salivarius, the mouthwash of the present invention can more effectively maintain the balance of oral microbial flora and promote oral health.

[0051] In one of the schemes, the particle size of the Lactobacillus salivarius microcapsules is 5-100 μm, preferably 10-80 μm, and more preferably 20-60 μm. The size of the particle size directly affects the dispersibility, stability and release characteristics of the microcapsules in the oral cavity. Within the above-mentioned particle size range, it is helpful to evenly disperse the microcapsules in water, avoid agglomeration, and ensure the uniform distribution of probiotics and effective contact with the oral mucosa. At the same time, the particle size also has a significant effect on the release rate and action time of probiotics. Within the above-mentioned particle size range, it is helpful to control the release, prolong the action time of probiotics in the oral cavity, and improve the health care effect of mouthwash. In addition, precise control of particle size also helps to improve the sensory experience of the product, avoid roughness or granularity, thereby improving the comfort and satisfaction of consumers.

[0052] The present invention also provides a method for preparing the mouthwash composition, comprising the following steps:

[0053] 1) Activating the strain to form a Lactobacillus salivarius culture solution;

[0054] 2) mixing the pH adjuster, stabilizer and preservative with the domiphene solution,

[0055] 3) under aseptic conditions, adding the Lactobacillus salivarius culture solution in step 1) to the domiphene mixed solution in step 2), and dispersing the mixture evenly at a low temperature of 0-10° C. to form the mouthwash composition, so that the content of Lactobacillus salivarius in the final composition is 10 7 Up to 10 11 CFU / mL, and the concentration of domiphene was 0.05wt% to 0.2wt%.

[0056] Furthermore, the method for preparing the mouthwash composition also includes a method for microencapsulating Lactobacillus salivarius, specifically, after step 1), further including the following steps:

[0057] a. suspending Lactobacillus salivarius in an aqueous solution containing a stabilizer to form an initial suspension;

[0058] b. adding an aqueous solution of microcapsule wall material to the initial suspension, and mixing to form a mixed suspension having a stabilizer and a wall material;

[0059] c. Prepare microcapsules by emulsification or spray drying,

[0060] And after Lactobacillus salivarius is microencapsulated, no stabilizer is added in step 2).

[0061] The mouthwash composition and the preparation method thereof provided by the present invention are described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention. In addition, the raw materials and experimental instruments used in the preferred embodiments of the present invention are all commercially available.

[0062] 1. Preparation of Mouthwash Compositions of Examples and Comparative Examples

[0063] Example 1

[0064] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0065] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:0.63, sodium carboxymethyl cellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution;

[0066] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0067] Lactobacillus salivarius 5×10 10 CFU;

[0068] Sodium dihydrogen phosphate 0.69 g;

[0069] Disodium hydrogen phosphate 0.63 g;

[0070] Domiphene 0.1 g;

[0071] Sodium carboxymethyl cellulose 0.08 g;

[0072] Methyl paraben 0.08 g;

[0073] Sodium fluoride 0.02 g;

[0074] Vitamin C 0.01g;

[0075] The balance is water, add to 100ml;

[0076] The pH value based on this mouthwash composition is 7.0.

[0077] Example 2

[0078] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0079] The Lactobacillus salivarius culture solution is added to the aqueous solution suspended in sodium carboxymethylcellulose to form a primary suspension;

[0080] Add 0.1 wt % gelatin aqueous solution to the primary suspension to form a mixed suspension, then heat to 40° C. and form an emulsion under high-speed stirring at 1000 rad / min;

[0081] Then the temperature was lowered to -5°C to solidify the gelatin to form microcapsules and freeze-dried to obtain dried Lactobacillus salivarius microcapsules with an average particle size of 25 μm;

[0082] dispersing the Lactobacillus salivarius microcapsules in water again to form an encapsulated Lactobacillus salivarius culture solution;

[0083] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:0.63, methyl parahydroxybenzoate, sodium fluoride, vitamin C and domiphene solution,

[0084] 3) Under sterile conditions, the encapsulated Lactobacillus salivarius culture solution in step 1) is added to the mixed solution of domiphene in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0085] Lactobacillus salivarius 5.5×10 10 CFU;

[0086] Gelatin 0.58g;

[0087] Sodium dihydrogen phosphate 0.69 g;

[0088] Disodium hydrogen phosphate 0.63 g;

[0089] Domiphene 0.1 g;

[0090] Xanthan gum 0.08g;

[0091] Methyl paraben 0.08 g;

[0092] Sodium fluoride 0.02 g;

[0093] Vitamin C 0.01g;

[0094] The balance is water, add to 100ml;

[0095] The pH value based on this mouthwash composition is 7.0.

[0096] Example 3

[0097] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0098] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:0.63, sodium carboxymethyl cellulose, methyl paraben, vitamin C and domiphene solution;

[0099] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0100] Lactobacillus salivarius 2×10 7 CFU;

[0101] Sodium dihydrogen phosphate 0.69 g;

[0102] Disodium hydrogen phosphate 0.63 g;

[0103] Domiphene 0.05 g;

[0104] Sodium carboxymethyl cellulose 0.05 g;

[0105] Methyl paraben 0.05 g;

[0106] Vitamin C 0.005g;

[0107] The balance is water, add to 100ml;

[0108] The pH value based on this mouthwash composition is 7.0.

[0109] Example 4

[0110] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0111] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:0.63, xanthan gum, phenoxyethanol, sodium fluoride and domiphene solution;

[0112] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0113] Lactobacillus salivarius 8×10 10 CFU;

[0114] Sodium dihydrogen phosphate 0.69 g;

[0115] Disodium hydrogen phosphate 0.63 g;

[0116] Domiphene 0.2 g;

[0117] Xanthan gum 0.1g;

[0118] Phenoxyethanol 0.08g;

[0119] Sodium fluoride 0.05 g;

[0120] The balance is water, add to 100ml;

[0121] The pH value based on this mouthwash composition is 7.0.

[0122] Example 5

[0123] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0124] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and sodium dihydrogen phosphate in a molar ratio of 1:0.41, sodium carboxymethyl cellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution;

[0125] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0126] Lactobacillus salivarius 5×10 10 CFU;

[0127] Sodium dihydrogen phosphate 0.69 g;

[0128] Disodium hydrogen phosphate 0.29 g;

[0129] Domiphene 0.1 g;

[0130] Sodium carboxymethyl cellulose 0.08 g;

[0131] Methyl paraben 0.08 g;

[0132] Sodium fluoride 0.02 g;

[0133] Vitamin C 0.01g;

[0134] The balance is water, add to 100ml;

[0135] The pH value based on this mouthwash composition was 6.8.

[0136] Example 6

[0137] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0138] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:1, sodium carboxymethyl cellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution;

[0139] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0140] Lactobacillus salivarius 5×10 10 CFU;

[0141] Sodium dihydrogen phosphate 0.69 g;

[0142] Disodium hydrogen phosphate 0.71 g;

[0143] Domiphene 0.1 g;

[0144] Sodium carboxymethyl cellulose 0.08 g;

[0145] Methyl paraben 0.08 g;

[0146] Sodium fluoride 0.02 g;

[0147] Vitamin C 0.01g;

[0148] The balance is water, add to 100ml;

[0149] The pH value based on this mouthwash composition was 7.2.

[0150] Comparative Example 1

[0151] Domiphene is mixed with sodium carboxymethylcellulose, methyl paraben, sodium fluoride and vitamin C to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0152] Domiphene 0.1 g;

[0153] Sodium carboxymethyl cellulose 0.08 g;

[0154] Methyl paraben 0.08 g;

[0155] Sodium fluoride 0.02 g;

[0156] Vitamin C 0.01g;

[0157] The balance is water, add to 100 ml.

[0158] Comparative Example 2

[0159] A mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:0.63, sodium carboxymethyl cellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution are mixed to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of each component are respectively:

[0160] Sodium dihydrogen phosphate 0.69 g;

[0161] Disodium hydrogen phosphate 0.63 g;

[0162] Domiphene 0.1 g;

[0163] Sodium carboxymethyl cellulose 0.08 g;

[0164] Methyl paraben 0.08 g;

[0165] Sodium fluoride 0.02 g;

[0166] Vitamin C 0.01g;

[0167] The balance is water, add to 100ml;

[0168] The pH value based on this mouthwash composition is 7.0.

[0169] Comparative Example 3

[0170] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0171] 2) mixing sodium carboxymethylcellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution;

[0172] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0173] Lactobacillus salivarius 5×10 10 CFU;

[0174] Domiphene 0.1 g;

[0175] Sodium carboxymethyl cellulose 0.08 g;

[0176] Methyl paraben 0.08 g;

[0177] Sodium fluoride 0.02 g;

[0178] Vitamin C 0.01g;

[0179] The balance is water, add to 100 ml.

[0180] Comparative Example 4

[0181] Compared with Example 1, other conditions remained unchanged, and the species of Lactobacillus was replaced by Lactobacillus plantarum from Lactobacillus salivarius.

[0182] Comparative Example 5

[0183] Compared with Example 1, other conditions remained unchanged, and the species of Lactobacillus was replaced by Lactobacillus casei from Lactobacillus salivarius.

[0184] Comparative Example 6

[0185] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0186] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:0.63, sodium carboxymethyl cellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution;

[0187] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0188] Lactobacillus salivarius 5×10 6 CFU;

[0189] Sodium dihydrogen phosphate 0.69 g;

[0190] Disodium hydrogen phosphate 0.63 g;

[0191] Domiphene 0.04 g;

[0192] Sodium carboxymethyl cellulose 0.08 g;

[0193] Methyl paraben 0.08 g;

[0194] Sodium fluoride 0.02 g;

[0195] Vitamin C 0.01g;

[0196] The balance is water, add to 100ml;

[0197] The pH value based on this mouthwash composition is 7.0.

[0198] Comparative Example 7

[0199] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0200] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:0.63, sodium carboxymethyl cellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution;

[0201] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0202] Lactobacillus salivarius 5×10 11 CFU;

[0203] Sodium dihydrogen phosphate 0.69 g;

[0204] Disodium hydrogen phosphate 0.63 g;

[0205] Domiphene 0.25 g;

[0206] Sodium carboxymethyl cellulose 0.08 g;

[0207] Methyl paraben 0.08 g;

[0208] Sodium fluoride 0.02 g;

[0209] Vitamin C 0.01g;

[0210] The balance is water, add to 100ml;

[0211] The pH value based on this mouthwash composition is 7.0.

[0212] Comparative Example 8

[0213] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0214] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:0.05, sodium carboxymethyl cellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution;

[0215] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0216] Lactobacillus salivarius 5×10 10 CFU;

[0217] Sodium dihydrogen phosphate 0.69 g;

[0218] Disodium hydrogen phosphate 0.035 g;

[0219] Domiphene 0.1 g;

[0220] Sodium carboxymethyl cellulose 0.08 g;

[0221] Methyl paraben 0.08 g;

[0222] Sodium fluoride 0.02 g;

[0223] Vitamin C 0.01g;

[0224] The balance is water, add to 100ml;

[0225] The pH value based on this mouthwash composition was 6.5.

[0226] Comparative Example 9

[0227] 1) taking out Lactobacillus salivarius from the strain preservation state (-80° C. refrigerator storage), inoculating the strain into a pre-prepared MRS medium under sterile conditions, raising the temperature to 30° C., and culturing overnight to form a Lactobacillus salivarius culture solution;

[0228] 2) mixing a mixed aqueous solution of sodium dihydrogen phosphate and disodium hydrogen phosphate in a molar ratio of 1:2, sodium carboxymethyl cellulose, methyl paraben, sodium fluoride, vitamin C and domiphene solution;

[0229] 3) Under sterile conditions, the Lactobacillus salivarius culture solution in step 1) is added to the domiphene mixed solution in step 2), and dispersed evenly at 5° C. to form the mouthwash composition. In the final composition, based on the total weight of the composition of 100 g (about 100 ml), the contents of the components are respectively:

[0230] Lactobacillus salivarius 5×10 10 CFU;

[0231] Sodium dihydrogen phosphate 0.69 g;

[0232] Disodium hydrogen phosphate 1.42 g;

[0233] Domiphene 0.1 g;

[0234] Sodium carboxymethyl cellulose 0.08 g;

[0235] Methyl paraben 0.08 g;

[0236] Sodium fluoride 0.02 g;

[0237] Vitamin C 0.01g;

[0238] The balance is water, add to 100ml;

[0239] The pH value based on this mouthwash composition is 7.5.

[0240] 2. In vitro simulated antibacterial test

[0241] Experimental methods:

[0242] Using soybean agar plates as culture media, culture mutans Streptococcus in an anaerobic environment at 37°C for 3 days. After the culture is completed, carefully pick the colonies with an inoculation loop and transfer them to the pre-prepared Todd Hewitt broth medium, and culture them under anaerobic conditions at 37°C for 18 hours. After that, take out 10mL of the culture solution and add it to a test tube containing 2mL Todd Hewitt broth medium (1% sucrose has been added to the medium). Place these test tubes at a 45° angle and continue to culture in an anaerobic environment at 37°C for 6 hours, during which a layer of biofilm forms on the inner wall of the test tube. After the culture is completed, remove the main culture medium in the test tube, and use 5mL of PBS buffer to gently rinse the biofilm on the inner wall of the test tube. The result is a model of simulated dental plaque for standby use.

[0243] 10 mL of each of the mouthwashes of Examples 1-6 and Comparative Examples 1-9 was taken and immersed in the dental plaque model and kept in a 37°C water bath for 3 minutes. Then the test liquid was poured out, 3 mL of PBS buffer was added to rinse the test tube twice, the liquid was poured out, 3 mL of sodium hydroxide (0.5 M) test solution was added, and the residual dental plaque was dispersed by vortex oscillator, and then the absorbance (550 nm) of the liquid in the test tube was detected.

[0244] Plaque removal rate = (absorbance of PBS buffer - absorbance of test group toothpaste supernatant) / absorbance of blank control

[0245] 3. In vitro simulated long-term antibacterial test

[0246] Using soybean agar plates as culture media, mutans Streptococcus was cultured in an anaerobic environment at 37°C for 3 days. After the culture is completed, carefully pick the colonies with an inoculation loop and transfer them to the pre-prepared Todd Hewitt broth medium, and culture them under anaerobic conditions at 37°C for 18 hours. Thereafter, take out 10 mL of the culture solution and add it to a test tube containing 2 mL of Todd Hewitt broth medium (to which 1 wt% maltooligosaccharide has been added). The test tubes were tilted at a 45° angle and continued to be cultured in an anaerobic environment at 37°C for 6 hours, during which a layer of biofilm formed on the inner wall of the test tube. After the culture is completed, the main culture medium in the test tube is removed, and the biofilm on the inner wall of the test tube is gently rinsed with 5 mL of PBS buffer. The result is a model for simulating dental plaque for standby use.

[0247] 10 mL of each of the mouthwashes of Examples 1-6 and Comparative Examples 1-9 was taken and immersed in the dental plaque model and kept in a 37°C water bath for 3 minutes. Then 10 ml of a mixture of glucoside amylase (0.1 wt%) and oligomaltosaccharide (1 wt%) was added to simulate the continuous decomposition process of starch in the oral cavity. The culture was continued for 24 h in a 37°C water bath, and then the test liquid was poured out, 3 mL of PBS buffer was added to rinse the test tube twice, the liquid was poured out, 3 mL of sodium hydroxide (0.5 M) test solution was added, and the residual dental plaque was dispersed by vortex oscillator, and then the absorbance (550 nm) of the liquid in the test tube was detected.

[0248] Plaque removal rate = (absorbance of PBS buffer - absorbance of test group toothpaste supernatant) / absorbance of blank control

[0249] Table 1 shows the immediate plaque removal rate (%) and 24h antibacterial effect of Examples 1-6 and Comparative Examples 1-9

[0250]

[0251]

[0252] It can be seen that the mouthwash compositions of Examples 1-6 can simultaneously satisfy the immediate antibacterial effect and the long-acting antibacterial effect, and the embodiment 2 in which Lactobacillus salivarius is microencapsulated has a relatively better antibacterial and long-acting antibacterial effect. It can be seen that the use of the mouthwash composition containing the present invention has played an inhibitory effect on harmful bacteria, thereby reducing dental plaque and preventing dental caries. Comparative Example 1 has only Domiphen as an antibacterial agent, Comparative Example 2 lacks Lactobacillus salivarius, Comparative Example 3 lacks a buffering agent for adjusting the pH value, and Comparative Examples 4 and 5 replace Lactobacillus salivarius with other lactobacilli, and the dual effect of antibacterial and long-acting antibacterial cannot be achieved. For the concentration range and pH control range, except that Comparative Example 7 can have the dual effect of antibacterial and long-acting antibacterial when the concentration range is large, the comparative examples of other situations do not have the dual effect of achieving antibacterial and long-acting antibacterial.

[0253] 4. Sensory Evaluation Test

[0254] The mouthwashes of Examples 1-6 and Comparative Examples 1-9 were subjected to sensory tests consisting of 8 test subjects in the test group. Each test subject was a healthy individual arranged on an objective basis according to the specified requirements. Sensory analysis was performed according to ISO 4121-2003 under the test conditions following ISO 8589, and 8 test subjects were counted to evaluate different samples.

[0255] Good - Positive comfort

[0256] Average - Can feel irritation in the mouth, but acceptable

[0257] Discomfort - obvious irritation, unacceptable

[0258] Table 2 shows the sensory test results of Examples 1-6 and Comparative Examples 1-9

[0259] sample good generally Discomfort Example 1 8 0 0 Example 2 7 1 0 Example 3 8 0 0 Example 4 7 1 0 Example 5 8 0 0 Example 6 7 1 0 Comparative Example 1 8 0 0 Comparative Example 2 8 0 0 Comparative Example 3 8 0 0 Comparative Example 4 8 0 0 Comparative Example 5 7 1 0 Comparative Example 6 8 0 0 Comparative Example 7 0 2 6 Comparative Example 8 5 3 0 Comparative Example 9 1 2 5

[0260] The results in Table 2 show that after excluding individual differences, the overall sensory perception of Examples 1-6 is good, while Comparative Examples 7 and 9, which have higher contents and overly alkaline pH, exhibit overall obvious sensory discomfort.

[0261] The above is only a preferred embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A mouthwash composition, characterized in that The mouthwash composition comprises the following active ingredients: Lactobacillus salivarius, domiphene, a pH regulator, a stabilizer and a preservative, wherein the content of Lactobacillus salivarius is 10 based on the total weight of the mouthwash composition. 7 Up to 10 11 CFU / mL, the concentration of domiphene is 0.05wt% to 0.2wt%, the pH adjuster adjusts the pH value of the mouthwash composition to 6.8 to 7.2, and the balance is water.

2. The mouthwash composition according to claim 1, characterized in that The pH regulator is selected from one or more of sodium dihydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium bicarbonate and sodium carbonate.

3. The mouthwash composition according to claim 1, characterized in that The concentration of the stabilizer is 0.05wt% to 0.1wt%, and is selected from one or more of sodium carboxymethyl cellulose, trehalose, xanthan gum, and polyvinyl pyrrolidone; and / or The concentration of the preservative is 0.05wt% to 0.1wt%, and the preservative is selected from one or more of methyl parahydroxybenzoate, propyl parahydroxybenzoate, phenoxyethanol, and nitrobenzoate.

4. The mouthwash composition according to claim 1, characterized in that The mouthwash composition also includes fluoride.

5. The mouthwash composition according to claim 1, characterized in that The mouthwash composition also includes vitamin C and / or vitamin E as antioxidants.

6. The mouthwash composition according to claim 1, characterized in that The Lactobacillus salivarius exists in the form of microcapsules.

7. The mouthwash composition according to claim 6, characterized in that The Lactobacillus salivarius is prepared in the form of microcapsules by comprising the following steps: a. suspending Lactobacillus salivarius in an aqueous solution containing a stabilizer to form an initial suspension; b. adding an aqueous solution of microcapsule wall material to the initial suspension, and mixing to form a mixed suspension having a stabilizer and a wall material; c. preparing microcapsules by emulsification or spray drying; The stabilizer is selected from one or more of trehalose, xanthan gum, sodium carboxymethyl cellulose or polyvinyl pyrrolidone; The microcapsule wall material is selected from one or more of gelatin, modified starch, cellulose derivatives or mixtures thereof.

8. The mouthwash composition according to claim 6, characterized in that The Lactobacillus salivarius exists in the form of microcapsules, and the particle size thereof is 5-500 μm.

9. A method for preparing the mouthwash composition according to claim 1, characterized in that: The steps include: 1) Activating the strain to form a Lactobacillus salivarius culture solution; 2) mixing the pH adjuster, stabilizer and preservative with the domiphene solution, 3) under aseptic conditions, adding the Lactobacillus salivarius culture solution in step 1) to the domiphene mixed solution in step 2), and dispersing the mixture evenly at a low temperature of 0-10° C. to form the mouthwash composition, so that the content of Lactobacillus salivarius in the final composition is 10 7 Up to 10 11 CFU / mL, and the concentration of domiphene was 0.05wt% to 0.2wt%.

10. The preparation method according to claim 9, characterized in that: The method further comprises the following steps after step 1): a. suspending Lactobacillus salivarius in an aqueous solution containing a stabilizer to form an initial suspension; b. adding an aqueous solution of microcapsule wall material to the initial suspension, and mixing to form a mixed suspension having a stabilizer and a wall material; c. Prepare microcapsules by emulsification or spray drying, And after Lactobacillus salivarius is microencapsulated, no stabilizer is added in step 2).

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