CN118105322A

CN118105322A - Toothpaste with antibacterial and hemostatic functions and preparation method thereof

Info

Publication number: CN118105322A
Application number: CN202410271506.XA
Authority: CN
Inventors: 温尧林; 邓小华; 邱一鸣; 朱凤圆; 冯滨
Original assignee: Tianjin Xinmanhe Biotechnology Co ltd
Current assignee: Tianjin Xinmanhe Biotechnology Co ltd
Priority date: 2024-03-11
Filing date: 2024-03-11
Publication date: 2024-05-31

Abstract

The invention provides toothpaste with antibacterial and hemostatic functions and a preparation method thereof, which belong to the technical field of oral cleaning products and comprise the following preparation raw materials in percentage by mass: 0.7-1.2% of collard leaf extract, 0.3-0.7% of perilla seed extract, 0.1-0.5% of pepper peel extract, 0.2-0.5% of ilex rotunda extract, 0.1-0.3% of jun-migratory fruit extract, 0.2-0.5% of silybum marianum fruit extract, 0.1-0.5% of mesona chinensis extract, 0.3-0.6% of myrobalan fruit extract, 1-4% of menthol, 20-25% of friction agent, 20-25% of wetting agent, 2-5% of foaming agent, 4-9% of tackifier, 3-7% of sodium ascorbyl phosphate, 4-8% of sodium hydroxybenzoate, 3-7% of potassium sorbate, 5-8% of trichlorogalactose, 1-2% of essence and the balance water. The toothpaste of the invention has remarkable antibacterial and hemostatic effects after being compounded by a plurality of components.

Description

Toothpaste with antibacterial and hemostatic functions and preparation method thereof

Technical Field

The invention belongs to the technical field of oral cleaning products, and particularly relates to toothpaste with antibacterial and hemostatic functions and a preparation method thereof.

Background

At present, toothpaste commonly used in the market is various in types, and the functional components of the toothpaste are divided according to the functional types, and generally comprise: anticaries agent, desensitizer, antibacterial hemostatic agent, stain removing and whitening agent, dental calculus inhibitor, breath freshener, etc. The antibacterial hemostatic toothpaste is a common functional product in the market, and can inhibit the growth of oral bacteria, inhibit dental plaque, or reduce gingivitis and bleeding and repair the functional components of oral mucosa through daily use. The effective component can be bisguanidine antibacterial agent (such as chlorhexidine), quaternary amine antibacterial agent (such as cetylpyridinium chloride), triclosan, allantoin, organic acid (such as tranexamic acid), phenolic antibacterial agent (such as eugenol, thymol, paeonol), plant extract, fluoride, etc. Plant extracts are increasingly favored by consumers as hemostatic and antibacterial active ingredients.

The Chinese patent publication No. CN117503672A discloses a natural herbal formula toothpaste with antibacterial effect, the main classification No. A61K8/00, and the toothpaste has the advantages of good antibacterial effect and the like by compounding various traditional Chinese medicine components, solves the problems that the existing toothpaste lacks antibacterial components, can not effectively inhibit the growth of bacteria in the oral cavity and can not inhibit the formation of dental plaque in the use process, and has poor inhibition effect on the bacteria; chinese patent publication No. CN117462471A discloses a multifunctional toothpaste with healthy components and a preparation method thereof, the main classification No. A61K8/00, and the prepared multifunctional toothpaste has scientific formula of healthy components, can resist bacteria and diminish inflammation, and can inhibit dental plaque and decayed tooth for a long time. However, the prior art still cannot fully meet the market demand, the antibacterial and hemostatic effects of active ingredients in the toothpaste with antibacterial and hemostatic functions of the plant extracts in the market are not ideal, the storability is poor, and the compatibility stability of the plant extracts in the toothpaste in the market and toothpaste matrixes is poor, so that quality problems such as water diversion, taste change and the like of the toothpaste occur in a short time, and the efficacy is seriously reduced or even fails.

Disclosure of Invention

The invention aims to provide toothpaste with antibacterial and hemostatic functions and a preparation method thereof, and the toothpaste has remarkable antibacterial and hemostatic effects and good stability, and does not change color, split water and become smell after long-time placement by adding the plant extract into the toothpaste.

In order to achieve the above object, the present invention provides the following technical solutions:

A toothpaste with antibacterial and hemostatic functions comprises the following preparation raw materials in percentage by mass: 0.7-1.2% of collard leaf extract, 0.3-0.7% of perilla seed extract, 0.1-0.5% of pepper peel extract, 0.2-0.5% of ilex rotunda extract, 0.1-0.3% of jun-migratory fruit extract, 0.2-0.5% of silybum marianum fruit extract, 0.1-0.5% of mesona chinensis extract, 0.3-0.6% of myrobalan fruit extract, 1-4% of menthol, 20-25% of friction agent, 20-25% of wetting agent, 2-5% of foaming agent, 4-9% of tackifier, 3-7% of sodium ascorbyl phosphate, 4-8% of sodium hydroxybenzoate, 3-7% of potassium sorbate, 5-8% of trichlorogalactose, 1-2% of essence and the balance water; wherein the pepper peel extract, the pubescent holly root extract, the jun migo fruit extract, the silybum marianum fruit extract, the mesona chinensis extract and the myrobalan fruit extract are prepared into a fermentation product A in advance, and then the fermentation product A is prepared by feeding materials; the collard leaf extract and the perilla seed extract are prepared into a fermentation product B in advance, and then the fermentation product B is fed into the toothpaste.

Further, the preparation method of the collard leaf extract comprises the following steps: (1) Soaking collard leaf in 3-5 times of water for 1-2 hr, distilling for 1-2 hr, and collecting liquid and solid substances respectively; (2) Soaking the solid in 6-8 times of water for 1-2 hr, reflux extracting for 2-3 times, reflux extracting for 0.5-1 hr each time, and collecting extractive solution; (3) Concentrating the liquid and the extractive solution at 60deg.C under reduced pressure to obtain collard leaf extract.

Further, the preparation method of the perilla seed extract comprises the following steps: superfine pulverizing fructus Perillae, selecting 5-20 μm powder, adding 6-7 times of water and 10-13wt% biological enzyme, performing enzymolysis at 40-45deg.C for 1-2 hr, centrifuging to obtain supernatant 1 and precipitate, adding the precipitate into 5-6 times of extractive solution composed of 35 deg.C water and ethanol at volume ratio of 1:1, adjusting pH to 4 with citric acid-sodium citrate buffer solution, extracting in an ultrasonic device, centrifuging to obtain supernatant 2, mixing supernatant 1 and supernatant 2, and drying to obtain fructus Perillae extract.

Mixing pericarpium Zanthoxyli extract, radix Ilicis Pubescentis extract, fructus Juglandis extract, herba Silybi Mariani extract, herba mesonae chinensis extract and fructus Chebulae extract, adding into fermentation broth A of 5-7 times mass, and performing aerobic fermentation at 32deg.C for 20-25 hr, wherein the fermentation broth A comprises the following components (by mass percent): glucose 1.5%, ammonium sulfate 0.1%, monopotassium phosphate 0.04%, magnesium sulfate 0.011% and water in balance; inoculating bacillus clausii, bacillus pseudofirmus and bacillus amyloliquefaciens with the mass ratio of 1:1-3:4-6, wherein the volume percentage of the inoculated bacillus clausii, the bacillus pseudofirmus and the bacillus amyloliquefaciens in the fermentation liquid A is 2.5%, and drying at 60 ℃ after the fermentation is finished to obtain a fermentation product A.

Mixing collard leaf extract and fructus Perillae extract, adding into fermentation liquid B5-7 times of the mass, and performing aerobic fermentation at 35deg.C for 25-30 hr, wherein the fermentation liquid B comprises the following components by mass percent: glucose 1%, ammonium sulfate 0.1%, monopotassium phosphate 0.05%, magnesium sulfate 0.012% and water in balance; and inoculating 3% of saccharomyces cerevisiae BY4742, nonomuria and Ji Shiku termates in the mass ratio of 0.1-0.5:2-4:1 in the fermentation liquid B, and drying at 60 ℃ after the compound fermentation is finished to obtain a fermentation product B.

Further, bacillus clausii accession no: BNCC191581; bacillus pseudofirmus accession No.: BNCC186420; bacillus amyloliquefaciens accession No.: BNCC336388; saccharomyces cerevisiae BY4742, accession number: BNCC342164. Nonomuria and accession number: BNCC276654; ji Shiku termyces, accession number: BNCC232040. All the strains are purchased from Beijing Beidou Nachuang biological technology institute.

The invention prepares the collard leaves by distillation and then extraction, simultaneously, in the treatment of the perilla seeds, superfine grinding is firstly carried out, which is beneficial to the enzymolysis of subsequent biological enzymes. The invention combines the extracts obtained by a specific method of the collard leaves and the perilla seeds to promote the synergistic interaction among the components, and uses specific strains to ferment, and the prepared fermentation product has good antibacterial and hemostatic effects when being added into the system of the invention. The commercially available two plant extracts have unstable content of active ingredients and may contain a large amount of other ingredients, and the substances have no antibacterial and hemostatic effects, but are possibly associated with the active ingredients to prevent the active ingredients from playing roles. However, other toothpaste matrices in the present system have poor compatibility with fermentation products of collard leaf extract and perilla seed extract.

Through a large number of research and development experiments, the inventor adds a plurality of plant extracts such as a pricklyash peel extract, a radix clerodendroa cyrtophylli extract, a junior fruit extract, a silybum marianum fruit extract, a mesona chinensis extract and a myrobalan fruit extract into the toothpaste, and through the addition of the extracts, the antibacterial and hemostatic functions of the toothpaste are improved by synergistic effect, and the inventor also surprisingly discovers that the compatibility of a toothpaste matrix, a collard leaf extract and a perilla seed extract is improved after the addition, so that the toothpaste is not easy to deteriorate at normal temperature, and the quality problems such as water diversion and taste change are not easy to occur. However, the stability of toothpaste at high temperatures is still not ideal.

The sources of the pepper peel extract, the ilex pubescens extract, the jun migo fruit extract, the silybum marianum fruit extract, the mesona chinensis extract and the myrobalan fruit extract are not particularly limited.

Further, the extraction conditions in the ultrasonic device are: under the action of 40Hz ultrasonic wave, the extraction temperature is 30 ℃ and the extraction time is 20-30min.

In order to improve the anti-inflammatory effect of the toothpaste, the biological enzyme comprises papain, ficin and bromelain with the mass ratio of 1-3:1:0.5-1.

Further, the wetting agent is one or more selected from sorbitol, glycerol and propylene glycol.

Further, the wetting agent is sorbitol, glycerol and propylene glycol with the mass ratio of 1:2-5:0.1-0.4.

The inventor adds a wetting agent to help dissolve other ingredients in the toothpaste, so that the ingredients are better and evenly mixed together, the uniform distribution of various active ingredients in the toothpaste is facilitated, and the consistent effect can be ensured for each use. The inventors have unexpectedly found that stability of the toothpaste at high temperatures can be improved when the humectant is sorbitol, glycerin and propylene glycol in a mass ratio of 1:2-5:0.1-0.4. It is hypothesized that the wetting agent under the conditions is favorable for maintaining the original structure and characteristics of the toothpaste, and meanwhile, a firm protective film can be formed on the surface of the active ingredient, so that the contact between the active ingredient and a high-temperature environment is reduced, and the stability of the active ingredient is improved.

Further, the friction agent is one or two selected from hydroxyapatite and hydrated silica.

Further, the tackifier is hydroxyethyl cellulose, xanthan gum and sodium carboxymethyl cellulose with the mass ratio of 2-5:1:0.5-1.

Compared with the antibacterial and hemostatic effects of chemical agents, the natural plant extract has the greatest defects that the natural plant extract is combined with the oral cavity for a short time, only about 2 minutes of tooth brushing is needed, and the active ingredients are flushed away after rinsing, so that the antibacterial and hemostatic effects are limited. In order to solve the contradiction, the inventor tries to adjust the auxiliary ingredients in the formula, and surprisingly discovers that when the tackifier is hydroxyethyl cellulose, xanthan gum and sodium carboxymethyl cellulose with the mass ratio of 2-5:1:0.5-1, the gum bleeding time can be reduced. The action mechanism of the tackifier does not uniformly improve the viscosity of paste, paste residues are not left in the oral cavity after the paste is rinsed by clean water, bacteria breeding in the oral cavity caused by the paste residues can be avoided, but the antibacterial and hemostatic effects are improved through tests, and the fact that the tackifier is used for enabling a fermentation product to be in contact with the inner wall of the oral cavity more easily after being combined is suspected, the retention time on the inner wall of the oral cavity is longer, the paste is not easy to wash away by back water, the action effect is better, and the hemostatic effect is improved.

Further, the foaming agent is selected from sodium lauroyl sarcosinate or sodium dodecyl sulfate.

Further, the pricklyash peel extract is xanthoxylin.

Further, the toothpaste comprises the following preparation raw materials in percentage by mass: 1% of collard leaf extract, 0.5% of perilla seed extract, 0.3% of pepper peel extract, 0.4% of ilex rotunda extract, 0.2% of junior fruit extract, 0.4% of silybum marianum fruit extract, 0.2% of mesona chinensis extract, 0.5% of myrobalan fruit extract, 2% of menthol, 22% of friction agent, 22% of wetting agent, 3% of foaming agent, 6% of tackifier, 5% of sodium ascorbyl phosphate, 6% of sodium hydroxybenzoate, 4% of potassium sorbate, 6% of trichlorogalactose, 1% of essence and the balance of water.

The invention also provides a preparation method of the toothpaste with the antibacterial and hemostatic functions, which comprises the following steps: dissolving fermentation product A, fermentation product B and menthol in water, adding friction agent, stirring for 10-20min, adding thickener, stirring for 5-10min, adding the rest components, stirring until no granule exists, and stirring under vacuum for 10-20min.

Further, the stirring speed is 900-1000r/min.

The raw materials of the present invention are all commercially available unless otherwise specified.

Compared with the prior art, the invention has the advantages that:

1. The invention provides toothpaste with antibacterial and hemostatic functions and a preparation method thereof, and the toothpaste has remarkable antibacterial and hemostatic effects and good stability by adding the plant extract into the toothpaste.

2. The invention distills and then extracts the collard leaves, superfine grinding is carried out on the perilla seeds, which is more beneficial to the enzymolysis of subsequent biological enzymes. The invention combines specific extracts and promotes the synergistic effect among the components, so that the system provided by the invention has good antibacterial and hemostatic effects by adding the collard leaf extract and the perilla seed extract.

3. The collard leaf extract and the perilla seed extract of the invention only use specific components, and the usage amount of the effective components is accurate and controllable, thereby improving the curative effect of bacteriostasis and hemostasis.

4. The toothpaste is added with various plant extracts including pepper peel extract, iron holly extract, jun migratory fruit extract, silybum marianum fruit extract, mesona chinensis extract and myrobalan fruit extract, so that the antibacterial and hemostatic functions of the toothpaste are improved, the compatibility of a toothpaste matrix, collard leaf extract and perilla seed extract is unexpectedly improved, and the toothpaste is not easy to deteriorate at normal temperature.

5. The wetting agent is added to help dissolve other ingredients in the toothpaste, so that the ingredients are better and evenly mixed together, the uniform distribution of various active ingredients in the toothpaste is facilitated, and the consistent effect can be ensured for each use. When the wetting agent is sorbitol, glycerol and propylene glycol with the mass ratio of 1:2-5:0.1-0.4, the stability of the toothpaste at high temperature can be improved.

Drawings

FIG. 1 is a graph showing the comparison of the inhibitory effect of sample B on Porphyromonas gingivalis growth;

FIG. 2 is a graph showing the experiment of inhibiting growth of Porphyromonas gingivalis in sample B;

FIG. 3 is a graph showing comparison of the effect of sample B on inhibition of F.nucleatum growth;

FIG. 4 is a graph showing the experiment of inhibiting the growth of F.nucleatum in sample B;

FIG. 5 is a graph showing comparison of the inhibition effect of sample B on the growth of Streptococcus mutans;

FIG. 6 is a graph of sample B showing inhibition of growth of Streptococcus mutans;

FIG. 7 is a graph showing comparison of the growth inhibitory effect of sample B on Streptococcus agalactiae;

FIG. 8 is a graph showing the experiment of inhibiting growth of Streptococcus gossypii in sample B;

FIG. 9 is a graph showing comparison of the growth inhibitory effect of sample B on Streptococcus distant;

FIG. 10 is a graph of sample B showing inhibition of growth of Streptococcus distant;

FIG. 11 is a graph showing the comparison of the growth inhibitory effect of the toothpaste prepared in example 1 and other toothpastes on Porphyromonas gingivalis;

FIG. 12 is a graph showing the experiment of the toothpaste prepared in example 1 and other toothpastes for inhibiting Porphyromonas gingivalis.

Detailed Description

The following description of the technical solutions in the embodiments of the present invention will be clear and complete, and it is obvious that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Example 1

The embodiment provides toothpaste with antibacterial and hemostatic functions, which comprises the following preparation raw materials in percentage by mass: 1% of collard leaf extract, 0.5% of perilla seed extract, 0.3% of pepper peel extract, 0.4% of ilex rotunda extract, 0.2% of junior fruit extract, 0.4% of silybum marianum fruit extract, 0.2% of mesona chinensis extract, 0.5% of myrobalan fruit extract, 2% of menthol, 22% of friction agent, 22% of wetting agent, 3% of foaming agent, 6% of tackifier, 5% of sodium ascorbyl phosphate, 6% of sodium hydroxybenzoate, 4% of potassium sorbate, 6% of trichlorogalactose, 1% of essence and the balance of water.

Preparing a fermentation product A from a pepper peel extract, a ilex pubescens extract, a monarch fruit extract, a silybum marianum fruit extract, a mesona chinensis extract and a myrobalan fruit extract in advance, and then preparing the toothpaste by feeding materials; preparing a fermentation product B from collard leaf extract and perilla seed extract in advance, and then feeding materials to prepare the toothpaste;

The preparation method of the collard leaf extract comprises the following steps: (1) Soaking collard leaves in 4 times of water for 1.5 hr, distilling for 1.5 hr, and collecting liquid and solid substances respectively; (2) Soaking the solid in 7 times of water for 1.5h, reflux-extracting for 3 times, reflux-extracting for 1h each time, and collecting extractive solution; (3) Concentrating the liquid and the extractive solution at 60deg.C under reduced pressure to obtain collard leaf extract.

The preparation method of the perilla seed extract comprises the following steps: superfine pulverizing fructus Perillae, selecting 5-20 μm powder, adding 7 times of water and 12wt% biological enzyme, performing enzymolysis at 43deg.C for 1 hr, centrifuging to obtain supernatant 1 and precipitate, adding the precipitate into 6 times of extractive solution composed of 35 deg.C water and ethanol at volume ratio of 1:1, extracting in ultrasonic device, centrifuging to obtain supernatant 2, mixing supernatant 1 and supernatant 2, and drying to obtain fructus Perillae extract.

The extraction conditions in the ultrasonic device are as follows: under the action of 40Hz ultrasonic wave, the extraction temperature is 30 ℃ and the extraction time is 25min.

The biological enzyme comprises papain, ficin and bromelain with the mass ratio of 2:1:0.7.

Mixing pericarpium Zanthoxyli extract, radix Ilicis Pubescentis extract, fructus Juglandis extract, herba Silybi Mariani extract, herba mesonae chinensis extract and fructus Chebulae extract, adding into 6 times mass of fermentation liquid A, and performing aerobic fermentation at 32deg.C for 22 hr, wherein the fermentation liquid A comprises the following components by mass percent: glucose 1.5%, ammonium sulfate 0.1%, monopotassium phosphate 0.04%, magnesium sulfate 0.011% and water in balance; inoculating bacillus clausii, bacillus pseudofirmus and bacillus amyloliquefaciens with the volume percentage of 2.5% in the fermentation liquid A, and drying at 60 ℃ after fermentation to obtain a fermentation product A;

Mixing collard leaf extract and fructus Perillae extract, adding into 6 times of fermentation liquid B, and performing aerobic fermentation at 35deg.C for 27 hr, wherein the fermentation liquid B comprises the following components by mass percent: glucose 1%, ammonium sulfate 0.1%, monopotassium phosphate 0.05%, magnesium sulfate 0.012% and water in balance; inoculating 3% of saccharomyces cerevisiae BY4742, nonomuria and Ji Shiku termates in the mass ratio of 0.3:3:1 in the fermentation liquid B, and drying at 60 ℃ after fermentation to obtain a fermentation product B.

The wetting agent is sorbitol, glycerol and propylene glycol with the mass ratio of 1:3:0.3.

The friction agent is prepared by compounding hydroxyapatite and hydrated silica according to a mass ratio of 3:1.

The tackifier is hydroxyethyl cellulose (purchased from merck, 09368), xanthan gum (purchased from merck, G1253) and sodium carboxymethyl cellulose (purchased from merck, C9481) with a mass ratio of 3:1:0.7. The foaming agent was sodium lauroyl sarcosinate (available from merck, Y0001772).

The embodiment also provides a preparation method of the toothpaste with antibacterial and hemostatic functions, which comprises the following steps: dissolving the fermentation product A, the fermentation product B and menthol in water, adding a friction agent, stirring for 15min at 950r/min, adding a thickening agent, stirring for 10min at 950r/min, continuously adding the rest components, stirring for 950r/min until no particles exist, and continuously stirring for 15min at 950r/min under vacuum.

Example 2

The differences between this embodiment and embodiment 1 are: a toothpaste with antibacterial and hemostatic functions comprises the following preparation raw materials in percentage by mass: 1.1% of collard leaf extract, 0.4% of perilla seed extract, 0.3% of pricklyash peel extract, 0.2% of ilex rotunda extract, 0.3% of Junzel fruit extract, 0.2% of silybum marianum fruit extract, 0.5% of mesona chinensis extract, 0.3% of myrobalan fruit extract, 2% of menthol, 24% of friction agent, 22% of wetting agent, 3% of foaming agent, 5% of tackifier, 4% of sodium ascorbyl phosphate, 6% of sodium hydroxybenzoate, 5% of potassium sorbate, 7% of trichlorogalactose, 1% of essence and the balance of water.

Example 3

The differences between this embodiment and embodiment 1 are: the biological enzyme comprises papain, ficin and bromelain in a mass ratio of 3:1:1. The wetting agent is sorbitol, glycerol and propylene glycol with the mass ratio of 1:5:0.2.

Comparative example 1

The difference between this comparative example and example 1 is: a toothpaste with antibacterial and hemostatic functions comprises the following preparation raw materials in percentage by mass: the composition comprises 0.4% of collard leaf extract, 1% of perilla seed extract, 0.7% of pepper peel extract, 0.1% of ilex rotunda extract, 0.5% of junior fruit extract, 0.1% of silybum marianum fruit extract, 0.2% of mesona chinensis extract, 0.3% of myrobalan fruit extract, 2% of menthol, 27% of friction agent, 23% of wetting agent, 5% of foaming agent, 3% of tackifier, 8% of sodium ascorbyl phosphate, 3% of sodium hydroxybenzoate, 5% of potassium sorbate, 7% of trichlorogalactose, 1% of essence and the balance of water.

Comparative example 2

The difference between this comparative example and example 1 is: a toothpaste with antibacterial and hemostatic functions comprises the following preparation raw materials in percentage by mass: 1% of collard leaf extract, 0.5% of perilla seed extract, 0.5% of pepper peel extract, 0.6% of ilex rotunda extract, 2% of menthol, 22% of friction agent, 22% of wetting agent, 3% of foaming agent, 6% of tackifier, 5% of sodium ascorbyl phosphate, 6% of sodium methylparaben, 4% of potassium sorbate, 6% of trichlorogalactose, 1% of essence and the balance of water.

Comparative example 3

The difference between this comparative example and example 1 is:

Inoculating bacillus subtilis (accession number: BNCC 188026), bacillus licheniformis (accession number: BNCC 336463) and bacillus pumilus (accession number: BNCC 103925) with a mass ratio of 1:1:1 in the fermentation broth A, and drying at 60 ℃ after fermentation to obtain a fermentation product A;

Inoculating bacillus subtilis (accession number: BNCC 188026), bacillus licheniformis (accession number: BNCC 336463) and bacillus pumilus (accession number: BNCC 103925) with the mass ratio of 1:1:1 in the fermentation broth B accounting for 3%, and drying at 60 ℃ after fermentation to obtain a fermentation product B.

All the strains are purchased from Beijing Beidou Nachuang biological technology institute.

Comparative example 4

The difference between this comparative example and example 1 is: the wetting agent is polyethylene glycol, glycerol and stearyl alcohol with the mass ratio of 1:1:1.

Comparative example 5

The difference between this comparative example and example 1 is: the tackifier is hydroxyethyl cellulose, carrageenan (purchased from merck, C1138) and colloidal magnesium aluminum silicate (CAS number: 71205-22-6) with the mass ratio of 1:1:1.

Performance testing

(1) Referring to GQT/ZY-DL-001, the instruction for the liver hemostasis test of rabbit,

1. Materials and methods

1. Test article toothpaste of examples 1-3 and comparative examples 1-5.

2. Experimental animals and raising environment

Common grade New Zealand white rabbits, female, weight 2.0kg ~2.5kg, by Guangdong province medical laboratory animal center (three water base) provides, laboratory animal production license number SCXK (Guangdong) 2019-0035, quality certification number No.44411600011769. Feed was provided by the medical laboratory animal center in Guangdong province. After purchasing, animals are fed in a single cage of a home animal house, and are used after quarantine is qualified, the use license number of experimental animals is SYXK (Yue) 2018-0137, the temperature of the animal house is 18-26 ℃, and the relative humidity is 40-70%.

3. Instrument and reagent

The main instrument is an electronic balance, the model number is QUINTIX3102-1CN, the stopwatch, and the model number is Tianfu brand PC70. Reagent 3% sodium pentobarbital.

4. The test method comprises the following steps:

4.1, taking 48 New Zealand white rabbits qualified in quarantine, and dividing the New Zealand white rabbits into 8 groups of 6 rabbits each.

4.2 After intravenous injection of 3% pentobarbital sodium for anesthesia, the liver was exposed by opening the abdomen layer by layer, and the wound surface of 1cm×1cm×3mm was clamped at the lower edge of the left lobe of the liver. After the wound surface is full of blood, 1.0g of the sample to be tested is coated on the whole wound surface, covered by gauze, and pressurized on the gauze by a weight of 50g, and the hemostatic effect is observed. The wounds of the model control group were not coated with the test sample, covered with gauze, and pressurized on the gauze with a 50g weight as a control. Every 30 seconds, observing bleeding of the wound, slightly dipping and sucking with a filter paper strip until the blood is no longer oozed, namely, the filter paper strip is no longer stained with blood, and recording the required time, namely, bleeding time.

4.3 Statistical analysis, experimental data are expressed in terms of standard deviation of mean soil, and SPSS software is adopted to conduct independent sample T test.

2. Test results

The test results of the local wound injury bleeding time test of the livers of the rabbits in each group are shown in the table 1 below.

Table 1: bleeding time test

As is clear from examples 1 and comparative example 5, the bleeding time becomes longer after changing the kind and the ratio of the thickener, indicating that the kind and the ratio of the specific thickener can reduce the bleeding time.

(2) Reference WS/T650-2019 methods for evaluating antibacterial and bacteriostatic effects; concentration of action the products of examples 1-3 and comparative examples 1-5 were as such and duration of action was 20min. The results of the antibacterial effect test are shown in table 2.

Table 2: antibacterial effect test

(3) The fermentation product B obtained by mixing the collard leaf extract and the perilla seed extract prepared in example 1 in a mass ratio of 2:1 was subjected to biological property test, and the results are shown in Table 3.

Table 3: testing of biological Properties

(4) Taking Porphyromonas gingivalis strains, and inoculating the Porphyromonas gingivalis strains on a blood agar plate for one-time activation. The other four strains were each subjected to activation culture on BHI plates. (2) after the cultivation is finished, the purity is determined by microscopic examination. Then respectively transferring BHI liquid culture medium for activating culture. (3) After microscopic examination is carried out to determine the purity, the BHI liquid culture medium is activated for 20 hours, transfer is carried out for secondary activation, activated culture solutions are respectively subjected to viable count, dilution factors with the concentration of the obtained bacterial suspension being 1-5 x 10 ⁶ cfu/ml are determined, the process is repeated for three times, and a bacterial suspension concentration inoculation amount scheme is confirmed.

Sample B aqueous solution preparation: the collard leaf extract and the perilla seed extract prepared in example 1 are mixed and fermented according to a mass ratio of 2:1 to obtain a fermentation product B, the fermentation product B is taken as a sample B to be dissolved in water to prepare 20mg/mL mother liquor, and the mother liquor is respectively diluted to prepare 10mg/mL,5mg/mL, 2.5mg/mL and 1.25mg/mL sample solutions, and the mixture is subjected to pasteurization at 70 ℃ for 30 minutes for later use.

Preparing toothpaste samples: weighing 3g of toothpaste prepared in example 1, dissolving in 7ml of water to prepare 30% toothpaste liquid, sufficiently shaking, soaking for 30 minutes, and centrifuging to obtain supernatant for later use; weighing 3g of silver-cleaning toothpaste, dissolving in 7ml of water to prepare 30% toothpaste liquid, sufficiently vibrating, soaking for 30 minutes, and centrifuging to obtain a supernatant for later use; 3g of Yunnan white drug powder toothpaste is weighed and dissolved in 7ml of water to prepare 30% toothpaste liquid, the toothpaste liquid is fully vibrated, soaked for 30 minutes, and the supernatant is centrifugally taken for standby.

Five concentration gradients of 10mg/mL,5mg/mL, 2.5mg/mL, 1.25mg/mL and 0.0625mg/mL are adopted to carry out bacteriostasis experiments on Porphyromonas gingivalis, and the bacteriostasis effect is determined. 1) Experimental group samples: 100ul of sample B+100ul of bacterial suspension are respectively packaged in an orifice plate, the concentration of the sample B is respectively 10mg/ml, 0.5mg/ml, 0.25mg/ml and 0.125 mg/ml+fresh bacterial suspension are respectively packaged in experimental group orifice plates with corresponding sample concentrations, and five parallel samples are respectively used for ensuring the bacterial concentration of each orifice plate to be consistent; 2) Positive control of control sample: 100ul of physiological saline and 100ul of bacterial-containing BHI culture medium are added, and three parallel samples are arranged; negative control: 100ul sample B (corresponding concentration) +100ul of sterile BHI medium, three replicates; the experimental group and the control group are simultaneously placed in an anaerobic environment at 37 ℃ for co-treatment for 24 hours, then an enzyme-labeled instrument is used for measuring the OD600 value of the pore plate, and the result is recorded by photographing;

Determination of the concentration of bacterial suspension according to the test procedure, each strain was subjected to secondary activation under appropriate conditions for 20 hours, and the test was repeated 3 times or more to obtain an initial bacterial suspension concentration of (1 to 5) ×10 ⁶ CFU/mL and the inoculum size thereof was as shown in table 4 below.

Table 4: inoculum size

1-12, The toothpaste prepared in example 1 has better inhibition effect on Porphyromonas gingivalis growth under the condition of multiple dilutions, and keeps consistent with the antibacterial result of the raw materials. The comparison experiment results of the toothpaste of the example 1 with the Yunnan white drug powder and the silver-cleaning toothpaste show that several toothpastes all show good growth inhibition effect, and only the toothpaste prepared in the example 1 has obvious inhibition effect on the growth of Porphyromonas gingivalis under the condition of multiple dilutions, and keeps consistent with the raw material antibacterial result.

(5) Stability test

The toothpastes prepared in examples 1 to 3 and comparative examples 1 to 5 were left at room temperature (25 ℃) and 40℃for 5 months, and the paste was observed. The results are shown in Table 5.

Table 5: stability test

As is clear from Table 5, the product of the present invention has good stability and does not deteriorate after long-term use. While the foregoing is directed to the preferred embodiments of the present invention, it will be appreciated by those skilled in the art that various modifications and adaptations can be made without departing from the principles of the present invention, and such modifications and adaptations are intended to be comprehended within the scope of the present invention.

Claims

1. The toothpaste with the antibacterial and hemostatic functions is characterized by comprising the following preparation raw materials in percentage by mass: 0.7-1.2% of collard leaf extract, 0.3-0.7% of perilla seed extract, 0.1-0.5% of pepper peel extract, 0.2-0.5% of ilex rotunda extract, 0.1-0.3% of jun-migratory fruit extract, 0.2-0.5% of silybum marianum fruit extract, 0.1-0.5% of mesona chinensis extract, 0.3-0.6% of myrobalan fruit extract, 1-4% of menthol, 20-25% of friction agent, 20-25% of wetting agent, 2-5% of foaming agent, 4-9% of tackifier, 3-7% of sodium ascorbyl phosphate, 4-8% of sodium hydroxybenzoate, 3-7% of potassium sorbate, 5-8% of trichlorogalactose, 1-2% of essence and the balance water;

wherein the pepper peel extract, the pubescent holly root extract, the jun migo fruit extract, the silybum marianum fruit extract, the mesona chinensis extract and the myrobalan fruit extract are prepared into a fermentation product A in advance, and then the fermentation product A is prepared by feeding materials;

The collard leaf extract and the perilla seed extract are prepared into a fermentation product B in advance, and then the fermentation product B is fed into the toothpaste;

The preparation method of the collard leaf extract comprises the following steps: (1) Soaking collard leaf in 3-5 times of water for 1-2 hr, distilling for 1-2 hr, and collecting liquid and solid substances respectively; (2) Soaking the solid in 6-8 times of water for 1-2 hr, reflux extracting for 2-3 times, reflux extracting for 0.5-1 hr each time, and collecting extractive solution; (3) Mixing the liquid and the extract, and concentrating at 60deg.C under reduced pressure to obtain collard leaf extract;

The preparation method of the perilla seed extract comprises the following steps: superfine pulverizing fructus Perillae, selecting 5-20 μm powder, adding 6-7 times of water and 10-13% of biological enzyme by mass percentage of fructus Perillae, performing enzymolysis at 40-45deg.C for 1-2 hr, centrifuging to obtain supernatant 1 and precipitate, adding the precipitate into 5-6 times of extractive solution composed of 35 deg.C water and ethanol at volume ratio of 1:1, extracting in ultrasonic device, centrifuging to obtain supernatant 2, mixing the supernatant 1 and supernatant 2, and drying to obtain fructus Perillae extract;

mixing pericarpium Zanthoxyli extract, radix Ilicis Pubescentis extract, fructus Juglandis extract, herba Silybi Mariani extract, herba mesonae chinensis extract and fructus Chebulae extract, adding into fermentation broth A of 5-7 times mass, and performing aerobic fermentation at 32deg.C for 20-25 hr, wherein the fermentation broth A comprises the following components (by mass percent): glucose 1.5%, ammonium sulfate 0.1%, monopotassium phosphate 0.04%, magnesium sulfate 0.011% and water in balance; inoculating bacillus clausii, bacillus pseudofirmus and bacillus amyloliquefaciens with the volume percentage of 2.5% in the fermentation liquid A in a mass ratio of 1:1-3:4-6, and drying at 60 ℃ after fermentation to obtain a fermentation product A;

Mixing collard leaf extract and fructus Perillae extract, adding into fermentation liquid B5-7 times of the mass, and performing aerobic fermentation at 35deg.C for 25-30 hr, wherein the fermentation liquid B comprises the following components by mass percent: glucose 1%, ammonium sulfate 0.1%, monopotassium phosphate 0.05%, magnesium sulfate 0.012% and water in balance; inoculating Saccharomyces cerevisiae BY4742, nonomuria and Ji Shiku termates with the volume percentage of 3% in the fermentation liquid B of 0.1-0.5:2-4:1, and drying at 60 ℃ after fermentation to obtain a fermentation product B.

2. The toothpaste with antibacterial and hemostatic functions according to claim 1, wherein the extraction conditions in the ultrasonic device are: under the action of 40Hz ultrasonic wave, the extraction temperature is 30 ℃ and the extraction time is 20-30min.

3. The toothpaste with antibacterial and hemostatic functions according to claim 2, wherein the biological enzymes comprise papain, ficin and bromelain in a mass ratio of 1-3:1:0.5-1.

4. The toothpaste having antibacterial and hemostatic functions according to claim 1, wherein the humectant is one or more selected from sorbitol, glycerin, propylene glycol.

5. The toothpaste with antibacterial and hemostatic functions according to claim 4, wherein the wetting agent is sorbitol, glycerol and propylene glycol in a mass ratio of 1:2-5:0.1-0.4.

6. The toothpaste having antibacterial and hemostatic functions according to claim 1, wherein the abrasive is one or both selected from the group consisting of hydroxyapatite and hydrated silica.

7. The toothpaste with antibacterial and hemostatic functions according to claim 1, wherein the tackifier is hydroxyethyl cellulose, xanthan gum and sodium carboxymethyl cellulose with a mass ratio of 2-5:1:0.5-1.

8. The toothpaste having antibacterial and hemostatic functions according to claim 1, wherein the foaming agent is selected from sodium lauroyl sarcosinate or sodium dodecyl sulfate.

9. The method for preparing the toothpaste with antibacterial and hemostatic functions according to any one of claims 1 to 8, comprising the following steps: dissolving fermentation product A, fermentation product B and menthol in water, adding friction agent, stirring for 10-20min, adding thickener, stirring for 5-10min, adding the rest components, stirring until no granule exists, and stirring under vacuum for 10-20min.

10. The method for preparing toothpaste with antibacterial and hemostatic functions according to claim 9, wherein the stirring speed is 900-1000r/min.