Traditional Chinese medicine probiotic synergistic antibacterial toothpaste based on porous mineral carrier and preparation method thereof

Abstract

The invention provides traditional Chinese medicine probiotic synergistic antibacterial toothpaste based on a porous mineral carrier and a preparation method of the traditional Chinese medicine probiotic synergistic antibacterial toothpaste, and belongs to the technical field of toothpaste. The preparation method comprises the following steps: firstly, obtaining a traditional Chinese medicine fermentation concentrated solution with an antibacterial function and probiotic metabolism freeze-dried powder, mixing to obtain a traditional Chinese medicine-probiotic mixed solution, loading the traditional Chinese medicine-probiotic mixed solution with perlite, and adding the loaded traditional Chinese medicine-probiotic mixed solution into a basic liquid phase for preparing the toothpaste. According to the preparation method, a chemical slow-release structure is replaced by porous perlite carrier adsorption, efficient utilization of active ingredients is achieved, pores are extruded through physical friction during tooth brushing, the active ingredients can be released to a dental plaque biofilm, and interference of a chemical slow-release material on the rheological stability of the paste is avoided. And meanwhile, the mineral carrier has the light friction characteristic, and the antibacterial aging is enhanced while the enamel abrasion is reduced.

Description

Technical Field

[0001] The present application relates to the technical field of antibacterial toothpaste, and in particular to a traditional Chinese medicine probiotic synergistic antibacterial toothpaste based on a porous mineral carrier and a preparation method thereof. Background Art

[0002] Bacteria and their metabolites in dental plaque are key contributors to oral diseases such as caries and periodontal disease, and mechanical toothbrushing is the primary means of daily plaque management. A wide variety of antibacterial toothpastes are currently available, with traditional Chinese medicine (TCM) toothpastes offering the highest safety. For example, mint, gallnut, and clove extracts have proven antimicrobial properties. However, traditional water-based extracts, due to their large molecular weight, have difficulty penetrating dental plaque biofilms, resulting in low efficacy. Furthermore, probiotic toothpastes often exhibit lamination or bloating due to low viable bacterial survival within the paste, uncontrolled metabolite release, and poor stability when coexisting with alkaline ingredients like baking soda. To improve the utilization of active ingredients, probiotic fermentation technology has been introduced into the field of TCM processing. By fermenting TCM formulas with specific strains, large active substances can be degraded into smaller compounds, significantly enhancing their ability to penetrate biofilms. Simultaneously, the fermentation process degrades potentially toxic components within the medicinal ingredients, achieving the dual goals of increasing efficacy and reducing toxicity.

[0003] However, the long-term antibacterial activity of fermentation products still relies on sustained-release carrier technology. Existing sustained-release systems often use polymer materials (such as chitosan nanofibers and sodium alginate microspheres). While these systems can prolong the duration of action, they are complex and costly to manufacture. Furthermore, high temperatures can easily cause the paste to gel, making it difficult to squeeze. Summary of the Invention

[0004] In order to solve at least one of the above technical problems, the technical solution adopted in this application is as follows.

[0005] The first aspect of the present application provides a method for preparing a traditional Chinese medicine probiotic synergistic antibacterial toothpaste based on a porous mineral carrier, comprising the following steps:

[0006] Preparation of Chinese herbal fermentation liquid: Mix Chinese gallnut and licorice in a ratio of 2-3:1, add 8-15 times water and boil for 45-90 minutes, filter and inoculate the filtrate with Lactobacillus salivarius, ferment and concentrate by ultrafiltration to obtain the Chinese herbal fermentation concentrate;

[0007] Preparation of probiotic metabolite freeze-dried powder: After anaerobic fermentation of Lactobacillus reuteri, the fermentation liquid after removing the bacteria is freeze-dried to obtain probiotic metabolite freeze-dried powder;

[0008] Preparation of a Chinese medicine-probiotics mixed solution: mixing the Chinese medicine fermentation concentrate and the probiotic metabolism freeze-dried powder in a ratio of 2 to 4:1 to obtain a Chinese medicine-probiotics mixed solution;

[0009] Carrier loading: mixing a porous mineral carrier and the traditional Chinese medicine-probiotic mixture in a ratio of 2 to 4:1, vacuum adsorption and thermal curing to obtain a carrier loaded with traditional Chinese medicine and probiotics, wherein the porous mineral carrier is perlite;

[0010] Preparation of toothpaste: adding the carrier loaded with traditional Chinese medicine and probiotics into 0.5 to 0.8 times the volume of the basic liquid phase to prepare the toothpaste, wherein the basic liquid phase includes a moisturizer.

[0011] In the present application, the main antibacterial component of the concentrated Chinese medicinal fermentation liquid obtained by fermenting gallnut and licorice with Lactobacillus salivarius is the small molecule active substance ellagic acid, and the content is relatively high.

[0012] In the present application, after Lactobacillus reuteri is fermented, the main metabolite is Reuterin antibacterial peptide.

[0013] In some embodiments of the present application, the Chinese medicine-probiotic mixture obtained by mixing the Chinese medicine fermentation concentrate and the probiotic metabolism freeze-dried powder has a dual antibacterial effect, and the antibacterial effect is synergistically enhanced.

[0014] In the present application, perlite in toothpaste not only has the function of an abrasive, but also has the function of a carrier, that is, it loads the Chinese medicine fermentation concentrate and the probiotic metabolism freeze-dried powder, and releases the adsorbed antibacterial active ingredients through physical squeezing when brushing teeth.

[0015] In some embodiments of the present application, the perlite has a particle size of 50-100 μm and a porosity greater than 80%.

[0016] Humectants are one of the main ingredients in toothpaste. They maintain the paste's softness and fluidity, making it easier to transport through pipes and process via filling. They also prevent the paste from drying out and hardening during its shelf life, preventing it from being squeezed out of a tube. They also lower the paste's freezing point, preventing it from losing its proper consistency and causing solid-liquid separation when thawed after freezing. In some embodiments of the present application, the humectant is selected from one or more of glycerin, sorbitol, propylene glycol, 1,2-hexanediol, butylene glycol, and polyethylene glycol.

[0017] An abrasive is a solid raw material component in toothpaste that works with a toothbrush to remove soft deposits on the tooth surface and reduce foreign matter such as stains, plaque, and tartar. In some embodiments of the present application, in addition to the perlite as an abrasive, after obtaining the carrier loaded with traditional Chinese medicine and probiotics, it is further mixed with one or more selected from hydrated silicon dioxide, calcium carbonate, calcium silicate, dibasic calcium phosphate dihydrate, calcium pyrophosphate, sodium metaphosphate, aluminum hydroxide, hydroxyapatite, and sodium bicarbonate.

[0018] Furthermore, the basic liquid phase contains a thickener, a foaming agent, a pH stabilizer and / or a flavor.

[0019] Thickeners, also known as adhesives or binders, are ingredients that disperse and swell in the toothpaste liquid phase, forming a stable colloid that suspends the toothpaste solid phase and prevents separation of the solid and liquid components. In some embodiments of the present application, the thickener is selected from one or more of sodium carboxymethylcellulose, hydroxyethylcellulose, carbomer, xanthan gum, hydroxypropyl guar gum, and carrageenan.

[0020] Foaming agents (surfactants) are ingredients in toothpaste that reduce liquid surface tension and provide excellent wetting, foaming, emulsification, and detergency. In some embodiments of the present application, the foaming agent is selected from one or more of sodium lauryl sulfate, sodium lauroyl sarcosinate, sodium palmitoyl glutamate, and cocamidopropyl betaine.

[0021] A pH stabilizer is a raw material component that helps maintain a stable pH during the shelf life of toothpaste to prevent water release and flatulence caused by the dissociation of the abrasive. In some embodiments of the present application, the pH stabilizer is selected from one or more of sodium benzoate, sodium pyrophosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, sodium bicarbonate, and sodium silicate.

[0022] Flavoring is typically one of the main ingredients in toothpaste, typically made from dozens of flavoring ingredients that meet food hygiene requirements. It imparts a cool, refreshing, and refreshing feeling to toothpaste, masking any unpleasant odors and flavors of other toothpaste ingredients without affecting its stability. In some embodiments of the present application, the flavor is selected from one or more of mint, spearmint, wintergreen, fruit, tea, and medicinal flavors.

[0023] Furthermore, the basic liquid phase also includes a taste improver, an appearance improver, a preservative, an antioxidant and / or an effective ingredient.

[0024] Taste modifiers are ingredients used to mask the unpleasant taste of certain toothpaste ingredients, imparting a pleasant flavor. Currently, taste modifiers primarily include sweeteners and salting agents. In some embodiments of the present application, the sweeteners include xylitol, sodium saccharin, aspartame, and sucralose. In some embodiments of the present application, the salting agent includes food-grade sodium chloride.

[0025] Appearance modifiers refer to raw materials and ingredients used to alter the appearance of toothpaste, creating a specific visual experience and functional associations, and are additives used when necessary. Currently used varieties include pigments, pigment lakes, and colored particles. In some embodiments of this application, the pigments include natural pigments (such as chlorophyll) and chemically synthesized pigments (such as titanium dioxide CI 77891, amaranth CI 16185, lemon yellow CI 19140, and brilliant blue CI 42090). These pigments can be used alone or in combination.

[0026] Preservatives are ingredients that prevent toothpaste from spoiling and deteriorating during its shelf life. Toothpaste contains multiple organic ingredients, undergoes complex production processes, and is susceptible to a wide variety of microorganisms. Therefore, the selection of toothpaste preservatives requires microbial challenge testing. Commonly used toothpaste preservatives include sodium benzoate and methylparaben (methylparaben). Active ingredients in toothpaste, such as chlorhexidine, cetylpyridinium chloride, triclosan, sodium fluoride, and sodium monofluorophosphate, also have preservative effects.

[0027] Antioxidants are ingredients used to prevent, inhibit, or delay discoloration and flavor changes caused by oxidation in toothpaste, thereby improving its stability. Antioxidants are added based on the specific formulation requirements. Examples of antioxidants used in toothpaste include butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), stannous chloride, phytic acid, ascorbic acid (vitamin C), and tocopherol (vitamin E).

[0028] Functional ingredients are raw materials that help functional toothpaste achieve one or more additional benefits beyond its basic functions. In some embodiments of this application, these functional ingredients include: mucosal repair agents, caries anticaries, desensitizers, antibacterial and anti-inflammatory hemostatics, ulcer removers and whitening agents, tartar inhibitors, and breath fresheners. In some specific embodiments of this application, the mucosal repair agent is zinc hyaluronate. Furthermore, a carrier loaded with traditional Chinese medicine and probiotics is mixed with zinc hyaluronate powder, and the pores of the carrier are used to absorb zinc ions.

[0029] The second aspect of the present application provides a perlite-based traditional Chinese medicine probiotic synergistic antibacterial toothpaste prepared by any preparation method described in the first aspect of the present application.

[0030] Compared with the prior art, this application has the following beneficial effects:

[0031] (1) Gallnut is a traditional Chinese medicine that is rich in gall tannins, which have antibacterial, anti-free radical, astringent, hemostatic, and analgesic effects. It helps reduce the accumulation of dental plaque and exerts its effects by inhibiting the growth and adhesion of Streptococcus mutans. It also helps to enhance the acid resistance of the dentin surface, inhibit caries, and promote remineralization after demineralization. Furthermore, the use of probiotic fermentation technology can improve the bioavailability of active ingredients: the extracellular enzymes secreted by Lactobacillus salivarius can decompose the gallnut cell wall, release small molecular active substances, and enhance their ability to penetrate the dental plaque biofilm; glycyrrhizin in licorice is fermented into glycyrrhetinic acid, which also significantly improves the anti-inflammatory activity and mucosal repair efficiency.

[0032] (2) Using Lactobacillus reuteri metabolites instead of live bacteria can improve the stability of the paste.

[0033] (3) The porous perlite carrier is used to absorb the active ingredients instead of the chemical sustained-release structure, achieving efficient utilization of the active ingredients. When brushing, the active ingredients are released into the dental plaque biofilm through physical friction and squeezing the pores, avoiding the interference of chemical sustained-release materials on the rheological stability of the paste. The mineral carrier has light friction properties, which reduces enamel wear while enhancing the antibacterial effect. The perlite carrier is compatible with existing toothpaste filling lines, and the modification only requires the addition of a vacuum adsorption module.

[0034] It should be understood that the content described in this section is not intended to identify the key or important features of the embodiments of the present application, nor is it intended to limit the scope of the present application. Other features of the present application will become easily understood through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The above and other objects, features and advantages of the exemplary embodiments of the present application will become readily understood by reading the detailed description below with reference to the accompanying drawings. In the accompanying drawings, several embodiments of the present application are shown in an illustrative and non-limiting manner, in which:

[0036] Figure 1 The figure shows a flow chart of the preparation method of the perlite-based traditional Chinese medicine probiotic synergistic antibacterial toothpaste in Example 1 of the present application. DETAILED DESCRIPTION

[0037] Unless otherwise indicated, implied from the context, or customary in the art, all parts and percentages in this application are based on weight, and the test and characterization methods used are current as of the filing date of this application. Where applicable, the contents of any patents, patent applications, or publications referred to in this application are incorporated herein by reference in their entirety, and their equivalent patent families are also incorporated by reference, particularly for definitions of relevant terms in the art disclosed therein. If the definition of a specific term disclosed in the prior art is inconsistent with any definition provided in this application, the definition of the term provided in this application shall prevail.

[0038] The numerical ranges in this application are approximate, so unless otherwise stated, they may include values outside the range. Numerical ranges include all values from the lower limit to the upper limit in increments of 1 unit, provided that there is an interval of at least 2 units between any lower value and any higher value. For ranges containing values less than 1 or containing fractions greater than 1 (e.g., 1.1, 1.5, etc.), 1 unit is appropriately considered to be 0.0001, 0.001, 0.01 or 0.1. For ranges containing single-digit numbers less than 10 (e.g., 1 to 5), 1 unit is generally considered to be 0.1. These are merely specific examples of what is intended to be expressed, and all possible combinations of values between the lowest and highest values listed are considered to be clearly recorded in this application.

[0039] The terms "comprising", "including", "having" and their derivatives do not exclude the presence of any other components, steps or processes and are irrelevant to whether these other components, steps or processes are disclosed in this application. To eliminate any doubt, all compositions using the terms "comprising", "including", or "having" in this application may include any additional additives, excipients or compounds unless expressly stated otherwise. In contrast, the term "essentially consisting of" excludes any other components, steps or processes from the scope of any subsequent description of the term, except those necessary for operational performance. The term "consisting of" does not include any components, steps or processes that are not specifically described or listed. Unless expressly stated otherwise, the term "or" refers to the listed members alone or in any combination thereof.

[0040] In order to make the technical problems, technical solutions and beneficial effects solved by the present application clearer and more understandable, the present application is further described in detail below in conjunction with the embodiments.

[0041] The following examples are provided herein to illustrate preferred embodiments of the present application. Those skilled in the art will appreciate that the techniques disclosed in the following examples represent techniques discovered by the inventors that can be used to implement the present application and, therefore, can be considered preferred embodiments of the present application. However, those skilled in the art will appreciate, based on this specification, that many modifications may be made to the specific embodiments disclosed herein while still achieving the same or similar results without departing from the spirit or scope of the present application.

[0042] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs, and the disclosures and materials cited therein are hereby incorporated by reference.

[0043] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many technical equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the claims.

[0044] The experimental methods in the following examples, unless otherwise specified, are all conventional methods. The instruments and equipment used in the following examples, unless otherwise specified, are all conventional laboratory instruments and equipment; the experimental materials used in the following examples, unless otherwise specified, are all purchased from conventional biochemical reagent stores.

[0045] Example 1 Preparation method of perlite-based traditional Chinese medicine probiotic synergistic antibacterial toothpaste

[0046] In this embodiment, the formula of the perlite-based traditional Chinese medicine probiotic synergistic antibacterial toothpaste is shown in Table 1.

[0047] Table 1: Perlite-based traditional Chinese medicine probiotic synergistic antibacterial toothpaste formula

[0048]

[0049] refer to Figure 1 The preparation method of perlite-based traditional Chinese medicine probiotic synergistic antibacterial toothpaste is as follows:

[0050] 1. Preparation of Traditional Chinese Medicine Fermentation Broth

[0051] Mix gallnut and liquorice in a ratio of 5:2, add 10 times the amount of water and boil for 1 hour, inoculate the filtrate with Lactobacillus salivarius CGMCC No.1.1881, ferment at 42°C for 72 hours, and concentrate by ultrafiltration to a specific gravity of 1.15 to obtain a Chinese medicine fermentation concentrate.

[0052] 2. Extraction of Probiotic Metabolites

[0053] Lactobacillus reuteri PB-LR09 (CCTCC No. M2018888) was anaerobically fermented in MRS medium at 37°C for 48 h, and the cells were removed by ultrafiltration using a 0.22 μm filter membrane. The cells were then freeze-dried to obtain probiotic metabolic freeze-dried powder (Reuterin powder).

[0054] 3. Carrier loading

[0055] The perlite was placed in a vacuum adsorption tank, and a mixture of traditional Chinese medicine fermentation concentrate and Reuterin powder (ratio 3:1) was injected. The mixture was adsorbed at -0.08 MPa for 30 minutes and then cured with hot air at 60°C for 2 hours. The cured drug-loaded mineral complex was dry-mixed with hyaluronic acid zinc powder, and zinc ions were adsorbed through the mineral pores.

[0056] 4. Basic liquid phase preparation

[0057] Add glycerin, sodium benzoate and xylitol to deionized water, heat to 40-45°C and stir until completely dissolved.

[0058] 5. Solid phase premixing and dispersion

[0059] Mix sodium carboxymethyl cellulose and the abrasive system (perlite + calcium silicate) by dry method, add slowly to the liquid phase in step 4, and stir under vacuum (-0.08 MPa) for 20 minutes until no particles are left.

[0060] 6. Surfactants and fragrances added (<40°C)

[0061] Add sodium lauroyl sarcosinate to the paste from step 5 and stir at low speed (300-400 rpm) for 10 minutes to avoid excessive foaming that may damage the paste structure. Add peppermint oil and continue stirring for 5 minutes until the aroma is evenly dispersed.

[0062] 7. Degassing and filling

[0063] The paste was transferred to a vacuum (-0.1 MPa) degassing tank for degassing for 15 minutes, filled into an aluminum tube and sealed, and then aged at a constant temperature of 25°C for 48 hours to stabilize the rheological properties.

[0064] Comparative Example 1: Porous mineral carrier replacement

[0065] The difference between Comparative Example 1 and Example 1 is that the porous mineral carrier is replaced by an equal amount of porous zeolite instead of perlite, and the other components and production steps remain unchanged.

[0066] Comparative Example 2: Live Bacteria Replacement

[0067] The difference between Comparative Example 2 and Example 1 is that the lyophilized powder of Lactobacillus reuteri metabolite is replaced with live Lactobacillus reuteri bacteria.

[0068] Comparative Example 3: Live Bacteria Replacement

[0069] The difference between Comparative Example 3 and Example 1 is that the freeze-dried powder of Lactobacillus reuteri metabolites is replaced with microspheres encapsulating live bacteria, that is, sodium alginate is used to encapsulate live Lactobacillus reuteri.

[0070] Example 2 Drug loading rate detection

[0071] In this study, liquid chromatography was used to detect the amount of traditional Chinese medicine and probiotics loaded in different carriers, and the effective antibacterial components, ellagic acid and Reuterin antimicrobial peptide (Reuterin), were detected.

[0072] Sample preparation: Take 0.5 g of drug-loaded perlite or zeolite sample, add 10 mL of acetonitrile-water-acetic acid mixture (volume ratio 20:4.95:0.05), and ultrasonically elute for 2 h; centrifuge (10000 rpm, 10 min), and take the supernatant to pass through a 0.45 μm organic filter membrane.

[0073] HPLC detection: The chromatographic conditions were a C18 column, a mobile phase of acetonitrile-0.1% phosphoric acid water (30:70), a flow rate of 1.0 mL / min, and detection wavelengths of 280 nm (ellagic acid) and 210 nm (Reuterin).

[0074] Calculate the content by standard curve method: drug loading rate (%) = (eluted drug mass / carrier mass) × 100%

[0075] The results are shown in Table 2.

[0076] Table 2: Drug loading rates of different carriers

[0077]

[0078] It can be seen that the drug loading rate of zeolite is significantly lower than that of perlite, with a decrease of about 15%~20%.

[0079] Example 3 Extrusion Release Rate Detection

[0080] Simulated extrusion: Mix the drug-loaded carrier with the abrasive (porous mineral carrier: calcium silicate = 3:1), place it in the RDA tester, apply 150g pressure, use a medium hardness brush, and oscillate for 2 hours.

[0081] Release amount determination: Collect the mixed solution, separate the solid residue by centrifugation, and detect the active ingredient content in the supernatant by HPLC.

[0082] Release rate (%) = (release amount / drug loading amount) × 100%

[0083] The results are shown in Table 3.

[0084] Table 3 Extrusion release rate of different carriers

[0085]

[0086] It can be seen that when using zeolite as a carrier, the extrusion release rate is less than 60% of that of perlite, and the residual rate is as high as 45%.

[0087] Example 4 Paste Stability Test

[0088] The toothpastes prepared in Example 1 and Comparative Examples 1 to 3 were placed in an environment of 40°C and 75% relative humidity for 3 months, and the appearance of the toothpastes and the presence of flatulence were observed. The results are shown in Table 4.

[0089] Table 4: Stability test results of different toothpastes

[0090]

[0091] Table 4 shows that the toothpaste using a porous perlite carrier and a freeze-dried form of probiotic metabolites achieved the best stability. Substituting perlite for zeolite significantly altered the toothpaste's appearance. Substituting live bacteria for freeze-dried Lactobacillus reuteri metabolites not only deteriorated the toothpaste's appearance but also resulted in severe water loss and flatulence. Even when microspheres encapsulated with live bacteria were used, the toothpaste exhibited a slight yellowing, as well as slight water loss and flatulence.

[0092] Example 5 Detection of pH Changes in Pastes

[0093] Sample preparation: The toothpastes prepared in Example 1 and Comparative Examples 2 to 3 were diluted with 9 mL of deionized water at a ratio of 1 g, shaken in a 40° C. water bath for 5 min, and cooled to 25° C.

[0094] pH determination: Calibrate the pH meter (pH 4.0 / 7.0 buffer), immerse the glass electrode in the sample, stir for 1 min, read the result, repeat 3 times and take the average value.

[0095] The test was performed once on day 0 and day 30 respectively. The results are shown in Table 5.

[0096] Table 5: pH test results of different toothpastes

[0097]

[0098] As shown in Table 5, the pH of the toothpaste prepared in Example 1 is relatively stable. Clinical studies have shown that a pH below 5.5 may corrode tooth enamel. In Comparative Example 2, the lyophilized powder of the Lactobacillus reuteri metabolite was replaced with live bacteria, causing the pH of the toothpaste to drop to 5.2 after 30 days, posing a clinical risk.

[0099] Example 6 Antibacterial effect detection

[0100] 1. Test strains

[0101] ① Streptococcus mutans (ATCC 25175): aerobic bacteria, the main pathogen of dental caries;

[0102] ② Porphyromonas gingivalis (ATCC 33277): an obligate anaerobe and the core pathogen of periodontitis.

[0103] 2. Culture medium

[0104] ①Streptococcus mutans: brain heart infusion agar (BHI agar);

[0105] ② Porphyromonas gingivalis: anaerobic agar (containing 5% defibrinated sheep blood).

[0106] 3. Main equipment

[0107] Biochemical incubator (37°C ± 1°C), anaerobic culture system (6% O2, 7.2% CO2, 7.2% H2, 79.7% N2), sterile filter paper (6 mm in diameter), colony counter (for measuring the diameter of the inhibition zone).

[0108] 4. Antibacterial Experiment

[0109] Step 1: Preparation of bacterial suspension

[0110] Streptococcus mutans: Inoculate activated bacteria on BHI agar, culture aerobically at 37°C for 48 hours, and dilute with sterile PBS to a concentration of (1.0-9.0) × 10 5 CFU / mL.

[0111] Porphyromonas gingivalis: Inoculate blood agar plates under anaerobic conditions, culture anaerobically at 37°C for 7 days, and dilute to the same concentration in PBS.

[0112] Step 2: Toothpaste sample processing

[0113] Instant antibacterial group: 1 g of the toothpaste solution prepared in Example 1 or Comparative Examples 1 to 3 was added to 9 mL of sterile saline and mixed by oscillation (200 rpm, 1 h) to prepare a 10% dilution.

[0114] Continuous antibacterial group: The toothpaste solutions were coated on glass slides and dried at 37°C for 4 hours to simulate the oral residual environment after brushing teeth, and then diluted with normal saline in the same way.

[0115] Step 3: Preparation of antibacterial tablets

[0116] Immerse the sterile filter paper in the toothpaste dilution for 5 seconds, remove it and place it in a sterile dish to dry at room temperature (to avoid volatilization of the ingredients).

[0117] Step 4: Antibacterial experiment operation

[0118] Apply bacterial solution: Use a spreader to evenly apply the bacterial suspension on the surface of the corresponding culture medium.

[0119] Place antibacterial sheets: Place 4 sets of filter paper sheets symmetrically on each dish:

[0120] ① Blank control (normal saline)

[0121] ② Positive control (0.2% chlorhexidine)

[0122] ③ Toothpaste samples in the immediate antibacterial group

[0123] ④ Toothpaste samples in the continuous antibacterial group

[0124] Gently press the filter paper to ensure that it adheres to the culture medium.

[0125] Culture and observation: Streptococcus mutans was cultured aerobically at 37℃ for 24 hours; Porphyromonas gingivalis was cultured anaerobically at 37℃ for 48 hours.

[0126] Step 5: Inhibition zone measurement

[0127] After the incubation period, the diameter of the inhibition zone was measured (accurate to 0.1 mm) using a colony counter. The results are shown in Table 6.

[0128] Table 6: Antibacterial effect of various toothpastes

[0129]

[0130] As can be seen from Table 6, the toothpaste prepared in Example 1 has a moderately sensitive antibacterial effect on the two core oral pathogens (>15 mm), and has a stronger inhibitory effect on Porphyromonas gingivalis; among them, the toothpaste prepared in Example 1 has a smaller inhibition zone in the continuous inhibition group but is still >15 mm (moderate inhibition), and the antibacterial activity attenuation rate is <12%, highlighting the advantages of the carrier technology.

[0131] The antibacterial effects of the comparative examples were significantly lower than those of the toothpaste in Example 1, and the antibacterial effect of the comparative example 2 in which live bacteria were directly added was the worst.

[0132] In summary, the toothpaste prepared in Example 1 has both immediate and strong antibacterial effect and sustained release ability, which meets the clinical needs of functional toothpaste.

[0133] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of this application, those skilled in the art may make various changes or modifications to this application, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A method for preparing a traditional Chinese medicine probiotic synergistic antibacterial toothpaste based on a porous mineral carrier, characterized in that: The following steps are involved: Preparation of Chinese herbal fermentation liquid: Mix Chinese gallnut and licorice in a ratio of 2-3:1, add 8-15 times water and boil for 45-90 minutes, filter and inoculate the filtrate with Lactobacillus salivarius, ferment and concentrate by ultrafiltration to obtain the Chinese herbal fermentation concentrate; Preparation of probiotic metabolite freeze-dried powder: After anaerobic fermentation of Lactobacillus reuteri, the fermentation liquid after removing the bacteria is freeze-dried to obtain probiotic metabolite freeze-dried powder; Preparation of a Chinese medicine-probiotics mixed solution: mixing the Chinese medicine fermentation concentrate and the probiotic metabolism freeze-dried powder in a ratio of 2 to 4:1 to obtain a Chinese medicine-probiotics mixed solution; Carrier loading: the porous mineral carrier and the traditional Chinese medicine-probiotic mixture are mixed in a ratio of 2 to 4:1, vacuum adsorbed and thermally solidified to obtain a carrier loaded with traditional Chinese medicine and probiotics, wherein the porous mineral carrier is perlite; Preparation of toothpaste: adding the carrier loaded with traditional Chinese medicine and probiotics into 0.5 to 0.8 times the volume of the basic liquid phase to prepare the toothpaste, wherein the basic liquid phase includes a moisturizer.

2. The preparation method according to claim 1, characterized in that The perlite has a particle size of 50-100 μm and a porosity greater than 80%.

3. The preparation method according to claim 1, characterized in that The moisturizing agent is selected from one or more of glycerin, sorbitol, propylene glycol, 1,2-hexanediol, butylene glycol, and polyethylene glycol.

4. The preparation method according to any one of claims 1 to 3, characterized in that In the carrier loading step, after obtaining the carrier loaded with traditional Chinese medicine and probiotics, it is further mixed with one or more selected from hydrated silicon dioxide, calcium carbonate, calcium silicate, dibasic calcium phosphate dihydrate, calcium pyrophosphate, sodium metaphosphate, aluminum hydroxide, hydroxyapatite and sodium bicarbonate.

5. The preparation method according to any one of claims 1 to 3, characterized in that The base liquid phase also includes a thickener, a foaming agent, a pH stabilizer and / or a flavor.

6. The preparation method according to claim 5, characterized in that The thickener is selected from one or more of sodium carboxymethyl cellulose, hydroxyethyl cellulose, carbomer, xanthan gum, hydroxypropyl guar gum and carrageenan.

7. The preparation method according to claim 5, characterized in that The foaming agent is selected from one or more of sodium lauryl sulfate, sodium lauroyl sarcosinate, sodium palmitoyl glutamate and cocamidopropyl betaine.

8. The preparation method according to claim 5, characterized in that The pH stabilizer is selected from one or more of sodium benzoate, sodium pyrophosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, sodium bicarbonate and sodium silicate.

9. The preparation method according to claim 5, characterized in that The flavor is selected from one or more of mint flavor, spearmint flavor, wintergreen flavor, fruit flavor, tea flavor and medicinal flavor.

10. The perlite-based traditional Chinese medicine probiotic synergistic antibacterial toothpaste prepared by the preparation method according to any one of claims 1 to 9.

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