CN114796044A - A Chinese medicinal compound toothpaste with antibacterial, antiinflammatory and immunity enhancing effects, and its preparation method - Google Patents
A Chinese medicinal compound toothpaste with antibacterial, antiinflammatory and immunity enhancing effects, and its preparation method Download PDFInfo
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Abstract
The invention discloses a Chinese herbal compound toothpaste with bacteriostasis, anti-inflammation and immunity improvement and a preparation method thereof, the Chinese herbal compound toothpaste is prepared by extracts of honeysuckle, corydalis bungeana, buffalo horn, red tuckahoe, forsythia, cortex moutan, rhizoma coptidis, selfheal and astragalus root and paste matrix of toothpaste; the preparation method of the toothpaste disclosed by the invention is characterized in that the traditional Chinese medicine combined extract is obtained by twice water boiling and once alcohol precipitation, and the toothpaste is further prepared by adding the toothpaste matrix, is simple in preparation method, stable in process, and capable of realizing large-scale production, cleaning and whitening teeth, keeping oral hygiene, refreshing breath, preventing dental caries, inhibiting bacteria and resisting inflammation, has the function of immunoregulation, and is suitable for daily use in the morning and at night.
Description
Technical Field
The invention relates to a traditional Chinese medicine compound toothpaste with functions of bacteriostasis, anti-inflammation and immunity improvement, and also relates to a preparation method thereof.
Background
The basis of daily oral health care is to prevent and inhibit oral diseases caused by oral bacteria breeding. Conventional biological or chemical agents used for daily bacteriostasis or sterilization pose a potential risk of drug resistance, and inflammation-induced red swelling and pain can cause extreme discomfort to the patient. The traditional Chinese medicine compound can play the drug effect in a multi-target and multi-way mode, the medicines are compatible with each other, the drug resistance and the discomfort of a patient can be reduced to a certain extent, and the traditional Chinese medicine compound has certain advantages in oral protection. However, the traditional Chinese medicine toothpaste in the market has a single type and function, and does not exert the synergistic effect generated by the mutual compatibility of traditional Chinese medicinal materials; the antibacterial active ingredients used by the toothpaste are not strong in pertinence and have broad-spectrum antibacterial property, and can inhibit nonpathogenic resident bacteria in the oral cavity, so that the immunity of the oral cavity is further reduced, and the self-healing capacity of each tissue to injury is reduced. In addition, the compound traditional Chinese medicine is only a plurality of single medicines which are simply spliced and stacked, a certain amount of clinical verification is not available, the support of the traditional Chinese medicine theory is not available, and the clinical curative effect of the compound traditional Chinese medicine is yet to be evaluated.
Disclosure of Invention
The purpose of the invention is as follows: aiming at the problems of single type function and broad-spectrum antibacterial damage to oral flora of the traditional Chinese medicine toothpaste in the prior art, the invention provides the traditional Chinese medicine compound toothpaste which is antibacterial and anti-inflammatory and can improve the immunity; also provides a preparation method of the traditional Chinese medicine compound toothpaste.
The technical scheme is as follows: the traditional Chinese medicine compound toothpaste is prepared from extracts of honeysuckle, bunge corydalis herb, buffalo horn, red tuckahoe, weeping forsythia, tree peony bark, coptis chinensis, selfheal and astragalus root and a toothpaste matrix; the traditional Chinese medicine extract comprises the following raw materials in parts by weight: 30-40 parts of honeysuckle, 30 parts of bunge corydalis herb, 15 parts of buffalo horn, 20 parts of red tuckahoe, 15-30 parts of fructus forsythiae, 15 parts of cortex moutan, 3 parts of rhizoma coptidis from Sichuan of China, 15 parts of selfheal and 15 parts of astragalus.
Preferably, the weight percentage of the extract is 0.1-1.5%, and the balance is toothpaste matrix; the weight ratio is 30 portions of honeysuckle, 30 portions of corydalis tuber, 15 portions of buffalo horn, 20 portions of red tuckahoe, 15 portions of forsythia, 15 portions of cortex moutan, 10 portions of rhizoma coptidis from Sichuan of China, 15 portions of selfheal and 15-20 portions of astragalus root.
Preferably, the humectant is one of glycerol, polyethylene glycol or sorbitol, the surfactant is sodium dodecyl sulfate or sodium lauroyl sarcosinate, the sweetener is xylitol or stevioside, the flavoring agent is one of menthol, grapefruit essence, jasmine essence or watermelon essence, the preservative is sodium benzoate or ethyl p-hydroxybenzoate, and the whitening agent is sodium pyrophosphate and sodium fluoride; the toothpaste matrix comprises the following components in percentage by weight: 12 to 35 percent of humectant, 0.8 to 2 percent of surfactant, 0.1 to 0.5 percent of sweetening agent, 0.5 to 1 percent of flavoring agent, 0.01 to 0.15 percent of preservative, 0.1 to 0.2 percent of sodium pyrophosphate, 0.5 to 0.15 percent of sodium fluoride and the balance of water.
The preparation method of the traditional Chinese medicine compound toothpaste comprises the following steps:
(1) adding cornu Bubali into 10-15 times of water, decocting to obtain mixed aqueous solution A;
(2) weighing honeysuckle, corydalis bungeana, poria cocos, fructus forsythiae, cortex moutan, coptis chinensis, selfheal and astragalus membranaceus, mixing the traditional Chinese medicine mixture with 6-8 times of water by mass of the traditional Chinese medicine mixture, adding the traditional Chinese medicine mixture and the water by mass of the traditional Chinese medicine mixture into the mixed aqueous solution A, boiling and extracting, and filtering to obtain liquid medicine;
(3) adding 6-8 times of water into the residue, boiling, extracting, filtering to obtain medicinal liquid, mixing the filtrates, concentrating under reduced pressure to obtain concentrated solution, adding ethanol into the concentrated solution, stirring, standing, filtering to remove precipitate to obtain ethanol precipitation filtrate;
(4) weighing the traditional Chinese medicine combined extract, and uniformly mixing the traditional Chinese medicine combined extract with a toothpaste matrix to obtain the traditional Chinese medicine compound toothpaste.
Preferably, ethanol is added into the concentrated solution in the step (2) to achieve the ethanol content of 40-60% in the concentrated solution.
Preferably, the ratio of the traditional Chinese medicine composition extract to the toothpaste base in the step (4) is 1: 49.
Experiments of the astragalus extract and the traditional Chinese medicine combined extract on lymphocyte proliferation are carried out by an MTT colorimetric method, the active ingredients in the astragalus can promote lymphocyte proliferation so as to achieve the effect of improving the autoimmunity, the effect that vital qi exists in the interior but exogenous pathogenic factors cannot dry is achieved, the resistance of an oral cavity to pathogenic microorganisms is enhanced, the effect is best when the compatibility ratio of the astragalus and the monarch drug honeysuckle in the honeysuckle detoxification decoction original formula in the combined extract is 1:2, and therefore, the astragalus is selected to be added in 15-20 parts;
at present, traditional Chinese medicine components are added into oral care products to act on oral cavity, and most of the oral care products still use traditional Chinese medicines to play the role of antibiosis and antiphlogosis, so that gingival bleeding is relieved, and recovery of oral ulcer is promoted. The lymph tissue around the oral cavity is distributed more, and the astragalus membranaceus is added to achieve a synergistic effect with the monarch drug honeysuckle in the honeysuckle detoxification decoction original formula, so that the effect of improving the autoimmunity is achieved.
Has the beneficial effects that: compared with the prior art, the invention has the following remarkable advantages:
the antibacterial anti-inflammatory traditional Chinese medicine compound toothpaste capable of improving immunity can clean and whiten teeth, keep oral hygiene, refresh breath, prevent decayed teeth, inhibit bacteria and resist inflammation, and has the function of immunoregulation; the traditional Chinese medicine mixture is subjected to twice water boiling and once alcohol precipitation to obtain the traditional Chinese medicine composition extract, the preparation method is simple, the process is stable, and the large-scale production can be realized.
Drawings
FIG. 1 is a diagram showing the proliferation activity of PBMC lymphocytes by the Chinese herbal compound composition of the present invention;
FIG. 2 is a diagram of the proliferation activity of PBMC lymphocytes by single components of Chinese herbs.
Detailed Description
The technical scheme of the invention is further explained by combining the attached drawings.
Example 1
The traditional Chinese medicine compound toothpaste comprises the following raw materials in parts by weight:
1% of traditional Chinese medicine composition extract, 20% of silicon dioxide, 60% of polyethylene glycol and glycerol, 1% of carboxymethyl cellulose and xanthan gum, 2.5% of sodium dodecyl sulfate, 0.5% of sodium benzoate, 0.5% of xylitol, 1% of stevioside, 0.5% of menthol and the balance of water. The traditional Chinese medicine extract comprises the following raw material medicines: 30 parts of honeysuckle, 30 parts of bunge corydalis herb, 15 parts of buffalo horn, 20 parts of red tuckahoe, 15 parts of weeping forsythia, 15 parts of cortex moutan, 10 parts of rhizoma coptidis from Sichuan of China, 15 parts of selfheal and 15 parts of astragalus root.
The preparation method of the traditional Chinese medicine compound toothpaste comprises the following steps:
(1) firstly weighing buffalo horn according to a proportion, then weighing honeysuckle, bunge corydalis herb, red poria, fructus forsythiae, cortex moutan, rhizoma coptidis, selfheal and astragalus root, mixing to obtain a traditional Chinese medicine mixture, adding the buffalo horn into water with the mass of 15 times of that of the buffalo horn, decocting for 15 minutes, adding the traditional Chinese medicine mixture into the mixture, and boiling and extracting for 45 minutes with the water with the mass of 7 times of that of the mixture; (2) filtering to obtain medicinal liquid, adding water 7 times of the total weight of the medicinal residues, boiling and extracting for 30 minutes, filtering to obtain medicinal liquid, mixing the filtrates obtained after 2 times of extraction, concentrating under reduced pressure to obtain concentrated solution, concentrating to crude drug amount of 1g/mL, adding ethanol into the concentrated solution to make alcohol content 40-60%, stirring, mixing, and standing for 24 hours; (3) filtering to remove precipitate to obtain alcohol precipitation filtrate as Chinese medicinal extract; (4) weighing a certain amount of Chinese medicinal composition extract, uniformly mixing the Chinese medicinal composition extract with the toothpaste matrix, and preparing the Chinese medicinal compound toothpaste by a conventional method.
Example 2
The traditional Chinese medicine compound toothpaste comprises the following raw materials in parts by weight:
1.5 percent of traditional Chinese medicine composition extract, 20 percent of silicon dioxide, 60 percent of polyethylene glycol and glycerin, 1 percent of carboxymethyl cellulose and xanthan gum, 2.5 percent of sodium dodecyl sulfate, 0.5 percent of sodium benzoate, 0.5 percent of xylitol, 1 percent of grapefruit essence and the balance of water. The traditional Chinese medicine extract comprises the following raw material medicines: 30 parts of honeysuckle, 30 parts of bunge corydalis herb, 15 parts of buffalo horn, 20 parts of red tuckahoe, 15 parts of weeping forsythia, 15 parts of cortex moutan, 10 parts of rhizoma coptidis from Sichuan of China, 15 parts of selfheal and 15 parts of astragalus root.
The preparation method of the traditional Chinese medicine compound toothpaste comprises the following steps:
(1) firstly weighing buffalo horn according to a proportion, then weighing honeysuckle, bunge corydalis herb, red poria, fructus forsythiae, cortex moutan, rhizoma coptidis, selfheal and astragalus root, mixing to obtain a traditional Chinese medicine mixture, adding the buffalo horn into water with the mass of 15 times of that of the buffalo horn, decocting for 15 minutes, adding the traditional Chinese medicine mixture into the mixture, and boiling and extracting for 45 minutes with the water with the mass of 7 times of that of the mixture; (2) filtering to obtain medicinal liquid, adding water 7 times of the total weight of the medicinal residues, boiling and extracting for 30 minutes, filtering to obtain medicinal liquid, mixing the filtrates obtained after 2 times of extraction, concentrating under reduced pressure to obtain concentrated solution, concentrating to crude drug amount of 1g/mL, adding ethanol into the concentrated solution to make alcohol content 40-60%, stirring, mixing, and standing for 24 hours; (3) filtering to remove precipitate to obtain alcohol precipitation filtrate as Chinese medicinal extract; (4) weighing a certain amount of Chinese medicinal composition extract, uniformly mixing the Chinese medicinal composition extract with the toothpaste matrix, and preparing the Chinese medicinal compound toothpaste by a conventional method.
Example 3
The traditional Chinese medicine compound toothpaste comprises the following raw materials in parts by weight:
2% of traditional Chinese medicine composition extract, 20% of silicon dioxide, 60% of polyethylene glycol and glycerin, 1% of carboxymethyl cellulose and xanthan gum, 2.5% of sodium dodecyl sulfate, 0.5% of sodium benzoate, 0.5% of xylitol and 1.5% of sodium lauroyl sarcosinate, 1% of jasmine essence and the balance. The traditional Chinese medicine extract comprises the following raw material medicines: 30 parts of honeysuckle, 30 parts of bunge corydalis herb, 15 parts of buffalo horn, 20 parts of red tuckahoe, 15 parts of weeping forsythia, 15 parts of cortex moutan, 10 parts of rhizoma coptidis from Sichuan of China, 15 parts of selfheal and 15 parts of astragalus root.
The preparation method of the traditional Chinese medicine compound toothpaste comprises the following steps:
(1) firstly weighing buffalo horn according to a proportion, then weighing honeysuckle, bunge corydalis herb, red poria, fructus forsythiae, cortex moutan, rhizoma coptidis, selfheal and astragalus root, mixing to obtain a traditional Chinese medicine mixture, adding the buffalo horn into water with the mass of 15 times of that of the buffalo horn, decocting for 15 minutes, adding the traditional Chinese medicine mixture into the mixture, and boiling and extracting for 45 minutes with the water with the mass of 7 times of that of the mixture; (2) filtering to obtain medicinal liquid, adding water 7 times of the total weight of the medicinal residues, boiling and extracting for 30 minutes, filtering to obtain medicinal liquid, mixing the filtrates obtained after 2 times of extraction, concentrating under reduced pressure to obtain concentrated solution, concentrating to crude drug amount of 1g/mL, adding ethanol into the concentrated solution to make alcohol content 40-60%, stirring, mixing, and standing for 24 hours; (3) filtering to remove precipitate to obtain alcohol precipitation filtrate as Chinese medicinal extract; (4) weighing a certain amount of Chinese medicinal composition extract, uniformly mixing the Chinese medicinal composition extract with the toothpaste matrix, and preparing the Chinese medicinal compound toothpaste by a conventional method.
Example 4
The traditional Chinese medicine compound toothpaste comprises the following raw materials in parts by weight:
2% of traditional Chinese medicine composition extract, 20% of silicon dioxide, 60% of polyethylene glycol and glycerol, 1% of carboxymethyl cellulose and xanthan gum, 2.5% of sodium dodecyl sulfate, 0.5% of sodium benzoate, 0.5% of stevioside, 1.5% of sodium lauroyl sarcosinate, 1% of watermelon essence and the balance of water. The traditional Chinese medicine extract comprises the following raw material medicines: 30 parts of honeysuckle, 30 parts of bunge corydalis herb, 15 parts of buffalo horn, 20 parts of red tuckahoe, 15 parts of weeping forsythia, 15 parts of cortex moutan, 10 parts of rhizoma coptidis from Sichuan of China, 15 parts of selfheal and 15 parts of astragalus root.
The preparation method of the traditional Chinese medicine compound toothpaste comprises the following steps:
(1) firstly weighing buffalo horn according to a proportion, then weighing honeysuckle, bunge corydalis herb, red poria, fructus forsythiae, cortex moutan, rhizoma coptidis, selfheal and astragalus root, mixing to obtain a traditional Chinese medicine mixture, adding the buffalo horn into water with the mass of 15 times of that of the buffalo horn, decocting for 15 minutes, adding the traditional Chinese medicine mixture into the mixture, and boiling and extracting for 45 minutes with the water with the mass of 7 times of that of the mixture; (2) filtering to obtain medicinal liquid, adding water 7 times of the total weight of the medicinal residues, boiling and extracting for 30 minutes, filtering to obtain medicinal liquid, mixing the filtrates obtained after 2 times of extraction, concentrating under reduced pressure to obtain concentrated solution, concentrating to crude drug amount of 1g/mL, adding ethanol into the concentrated solution to make alcohol content 40-60%, stirring, mixing, and standing for 24 hours; (3) filtering to remove precipitate to obtain alcohol precipitation filtrate as Chinese medicinal extract; (4) weighing a certain amount of Chinese medicinal composition extract, uniformly mixing the Chinese medicinal composition extract with the toothpaste matrix, and preparing the Chinese medicinal compound toothpaste by a conventional method.
Example 5
The traditional Chinese medicine compound toothpaste comprises the following raw materials in parts by weight:
2% of traditional Chinese medicine composition extract, 20% of silicon dioxide, 60% of polyethylene glycol and glycerin, 1% of carboxymethyl cellulose and xanthan gum, 2.5% of sodium dodecyl sulfate, 0.5% of sodium benzoate, 0.5% of stevioside and 1.5% of sodium lauroyl sarcosinate, 1% of jasmine essence and the balance of water. The traditional Chinese medicine extract comprises the following raw material medicines: 30 parts of honeysuckle, 30 parts of bunge corydalis herb, 15 parts of buffalo horn, 20 parts of red tuckahoe, 15 parts of weeping forsythia, 15 parts of cortex moutan, 10 parts of rhizoma coptidis from Sichuan of China, 15 parts of selfheal and 15 parts of astragalus root.
The preparation method of the traditional Chinese medicine compound toothpaste comprises the following steps:
(1) firstly weighing buffalo horn according to a proportion, then weighing honeysuckle, bunge corydalis herb, red poria, fructus forsythiae, cortex moutan, rhizoma coptidis, selfheal and astragalus root, mixing to obtain a traditional Chinese medicine mixture, adding the buffalo horn into water with the mass of 15 times of that of the buffalo horn, decocting for 15 minutes, adding the traditional Chinese medicine mixture into the mixture, and boiling and extracting for 45 minutes with the water with the mass of 7 times of that of the mixture; (2) filtering to obtain medicinal liquid, adding water 7 times of the total weight of the medicinal residues, boiling and extracting for 30 minutes, filtering to obtain medicinal liquid, mixing the filtrates obtained after 2 times of extraction, concentrating under reduced pressure to obtain concentrated solution, concentrating to crude drug amount of 1g/mL, adding ethanol into the concentrated solution to make alcohol content 40-60%, stirring, mixing, and standing for 24 hours; (3) filtering to remove precipitate to obtain alcohol precipitation filtrate as Chinese medicinal extract; (4) weighing a certain amount of Chinese medicinal composition extract, uniformly mixing the Chinese medicinal composition extract with the toothpaste matrix, and preparing the Chinese medicinal compound toothpaste by a conventional method.
Antibacterial test (the bacteria used are all international standard strains)
Test (diffusion method-filter paper method)
Control group: gentamicin, clindamycin, terbinafine, and the same type of toothpaste sold in the market
Taking 4 groups (group A, group B, group C and group D) of culture dishes, wherein each group comprises 10 culture dishes, uniformly mixing a solid culture medium with the 4 bacterial solutions while the solid culture medium is hot, uniformly paving the culture dishes, and cooling (group A (inoculated with staphylococcus aureus) and clindamycin control, group B (inoculated with escherichia coli) and gentamicin control, group C (inoculated with bacillus subtilis) and gentamicin control, and group D (inoculated with candida albicans) and terbinafine control) to obtain a culture dish;
weighing 5mL of the traditional Chinese medicine combined extract prepared in the embodiment 1-5, the control group antibiotics, the commercially available toothpaste extract of the same type and blank control, respectively soaking filter paper sheets in the mixture for 2 hours, and uniformly and symmetrically attaching 4 small round filter paper sheets in each culture dish;
after the culture dishes of group a, group B, and group C to which the filter paper sheets were attached were placed in a constant temperature incubator at 37 ℃ for 24 hours and the culture dish of group D to which the filter paper sheets were attached was placed in a constant temperature incubator at 28 ℃ for 24 hours, the diameters of the four quasi-circular antibacterial areas in each dish were measured and averaged.
Taking 4 groups (group A, group B, group C and group D) of culture dishes, wherein 10 culture dishes are used in each group, uniformly mixing a solid culture medium with the 4 bacterial solutions while the solid culture medium is hot, uniformly paving the culture dishes, and cooling (group A (inoculated with staphylococcus aureus) and clindamycin control, group B (inoculated with escherichia coli) and gentamicin control, group C (inoculated with bacillus subtilis) and gentamicin control, and group D (inoculated with candida albicans) and terbinafine control) to obtain a culture dish;
weighing 5mL of the traditional Chinese medicine compound toothpaste extract prepared in the embodiment 1-5, the control group antibiotics, the commercially available toothpaste extract of the same type and blank control, respectively soaking filter paper sheets in the traditional Chinese medicine compound toothpaste extract for 2 hours, and uniformly and symmetrically attaching 4 small round filter paper sheets in each culture dish; after the group a, group B, and group C culture dishes to which the filter paper sheets were attached were placed upside down in a 37 ℃ constant temperature incubator and cultured for 24 hours, and the group D culture dishes to which the filter paper sheets were attached were placed upside down in a 28 ℃ constant temperature incubator and cultured for 24 hours, the diameters of the four quasi-circular antibacterial areas in each dish were measured and averaged.
Test (diffusion method-punching method)
Taking 4 groups (group A, group B, group C and group D) of culture dishes, wherein each group comprises 10 culture dishes, uniformly mixing a solid culture medium with the 4 bacterial solutions while the solid culture medium is hot, uniformly paving the culture dishes, cooling (group A (inoculated with staphylococcus aureus) and clindamycin contrast, group B (inoculated with escherichia coli) and gentamicin contrast, group C (inoculated with bacillus subtilis) and gentamicin contrast, and group D (inoculated with candida albicans) and terbinafine contrast), and uniformly and symmetrically punching 4 culture dishes after cooling to obtain culture dishes;
the same amount of liquid medicine (herbal extract prepared in examples 1-5, control antibiotics, commercial toothpaste extract of the same type, and blank control) was injected into each well of each culture dish;
the group a, group B and group C culture dishes with the liquid medicine injected into the small holes are placed in a constant temperature incubator at 37 ℃ for 24 hours, and the group D culture dishes with the liquid medicine injected into the small holes are placed in a constant temperature incubator at 28 ℃ for 24 hours, and then the diameters of the four quasi-circular antibacterial areas in each culture dish are measured and averaged.
Taking 4 groups (group A, group B, group C and group D) of culture dishes, wherein each group comprises 10 culture dishes, uniformly mixing a solid culture medium with the 4 bacterial solutions while the solid culture medium is hot, uniformly paving the culture dishes, cooling (group A (inoculated with staphylococcus aureus) and clindamycin contrast, group B (inoculated with escherichia coli) and gentamicin contrast, group C (inoculated with bacillus subtilis) and gentamicin contrast, and group D (inoculated with candida albicans) and terbinafine contrast), and uniformly and symmetrically punching 4 culture dishes after cooling to obtain culture dishes;
injecting equal amount of liquid medicine (the Chinese medicinal compound toothpaste extract prepared in examples 1-5, control group antibiotics, commercial toothpaste extract of the same type and blank control) into each culture dish;
the group a, group B and group C culture dishes with the liquid medicine injected into the small holes are placed in a constant temperature incubator at 37 ℃ for 24 hours, and the group D culture dishes with the liquid medicine injected into the small holes are placed in a constant temperature incubator at 28 ℃ for 24 hours, and then the diameters of the four quasi-circular antibacterial areas in each culture dish are measured and averaged.
And (3) performing an antibacterial test by adopting a diffusion method, if the corresponding sample has antibacterial property, displaying an antibacterial area on the culture medium, evaluating the antibacterial property of the sample according to the diameter of the antibacterial area, wherein the larger the diameter is, the stronger the antibacterial property is. The staphylococcus aureus, the candida albicans, the bacillus subtilis and the escherichia coli are common oral bacteria, the staphylococcus aureus is a gram-positive coccus which is often piled into a grape string shape and is easy to cause oral inflammation, the bacillus subtilis and the escherichia coli are common gram-negative bacteria, and the candida albicans is a fungus which is usually present in the oral cavity and the upper respiratory tract of a normal person and is easy to invade cells to cause diseases when the immune function of an organism is reduced.
Determination of minimum inhibitory and bactericidal concentrations
The bacterial species: staphylococcus aureus, escherichia coli, bacillus subtilis and candida albicans;
test (two-fold dilution method)
11 sterile 1.5mL centrifuge tubes were prepared, labeled, 100. mu.L of the herbal extract (the herbal extract prepared in examples 1-5) was added to centrifuge tube No. 1, mixed well with a pipette, 100. mu.L of the herbal extract was aspirated and added to centrifuge tube No. 2, and so on, to achieve concentrations of each herbal extract of 500.00, 250.00, 125.00, 62.50, 31.25, 15.62, 7.81, 3.90, 1.95, and 0.97. mu.g/L, respectively. No. 11 centrifuge tube was not added with drug as positive control;
preparing 11 large-size centrifuge tubes, labeling, respectively adding 40 μ L of Chinese medicinal extract with corresponding sequence number and 4ml of culture medium, mixing, pouring onto flat plate while hot, cooling, and inoculating 10 μ L of bacterial liquid, group A (inoculated with Staphylococcus aureus), group B (inoculated with Escherichia coli), and group C (inoculated with Bacillus subtilis) into three regions of each culture dish;
and (3) putting the culture dish into a constant-temperature biochemical incubator at 37 ℃ for 24 hours, and recording the minimum bacteriostatic concentration and the minimum bactericidal concentration.
TABLE 1 antibacterial experiment results of perforation method with formula ratio adjustment
As shown in table 1, the more the orange is, the larger the average value of the bacteriostatic circle is, the better the antibacterial effect is; the concentration of the traditional Chinese medicine compound composition is tentatively determined to be 0.5g/mL through a pre-experiment for testing. The other medicines including 30 parts of corydalis bungeana, 20 parts of red tuckahoe, 15 parts of cortex moutan and 15 parts of selfheal are added into each group of the traditional Chinese medicine compound composition. From table 1, it can be seen that: the traditional Chinese medicine compound composition comprises the following components: 30-40 parts of honeysuckle, 30 parts of bunge corydalis herb, 15 parts of buffalo horn, 20 parts of red tuckahoe, 15-30 parts of weeping forsythia, 15 parts of cortex moutan, 3 parts of rhizoma coptidis from Sichuan of China, 15 parts of selfheal and 15 parts of astragalus root, has the most obvious bacteriostatic effect, has the inhibitory effect on staphylococcus aureus, escherichia coli and bacillus subtilis, and has no inhibitory effect on candida albicans.
Lymphocyte proliferation assay
Taking out mouse spleen under aseptic condition, separating lymphocyte with lymphocyte separation liquid, and adjusting cell concentration to 1 × 10 with incomplete 1640 culture liquid 5 The volume is/mL for standby;
add mouse spleen lymphocyte suspension (1X 10) to 96-well culture plate 5 mL) 100. mu.L/well, control group 100. mu.L/well saline. The experimental group was added with 100 μ L of the Chinese medicinal composition extract prepared in examples 1-5 in sequence; gently mixing, and placing in 5% CO 2 Culturing in an incubator at 37 ℃ for 24 hours, adding 40mL of MTT solution into each hole under aseptic conditions, slightly mixing, continuing culturing for 4 hours, putting into a centrifuge for 1500r/min, centrifuging for 10 minutes, removing supernatant by using filter paper, adding 150 mu L of dimethyl sulfoxide into each hole, standing at room temperature for 15 minutes, and measuring the absorbance A value of each hole;
data processing
The cell proliferation promoting rate was calculated as follows:
cell proliferation promoting rate (A) Experiment of -A Control )/A Experiment of ×100％
FIG. 1 shows the effect of the Chinese herbal compound composition on the proliferation activity of PBMC lymphocytes, and FIG. 2 shows the effect of the Chinese herbal single component on the proliferation activity of PBMC lymphocytes.
The traditional Chinese medicine components are a complex system, lymphocyte proliferation test detection is firstly carried out on the total extracts of various chemical components of astragalus membranaceus, and the results show that, compared with a blank control group, the cell activity can be enhanced by 0-25 mg/ml of total astragalus saponin extract, the cell activity can be enhanced by 0-50 mg/ml of total astragalus flavone extract, and the cell activity can be enhanced by 0-200 mg/ml of total astragalus polysaccharide extract, and experiments prove that the saponin, flavone and polysaccharide components of the astragalus membranaceus have promotion effects on lymphocyte proliferation within a certain concentration range. Secondly, the lymphocyte proliferation test detection is carried out on the honeysuckle and forsythia aqueous extracts as the monarch drugs and the monarch drugs in the prescription, and the result shows that compared with a blank control group, the 25-200 mg/ml forsythia aqueous extract can enhance the cell activity, the 0-12.5 mg/ml forsythia aqueous extract can enhance the cell activity, and the experiment proves that the honeysuckle and forsythia aqueous extracts have the promotion effect on the lymphocyte proliferation within a certain concentration range. Fig. 1 shows that, compared with a blank control group, the results of the traditional Chinese medicine composition extract in an amount of 0.25-1 mg/ml can enhance the cell activity, and the activity enhancement shows an ascending trend, so that lymphocyte proliferation test detection performed on the concentration of the traditional Chinese medicine composition extract has a significant promoting effect on lymphocyte proliferation.
Claims (7)
1. A Chinese medicinal compound toothpaste with antibacterial, antiinflammatory and immunity enhancing effects is prepared from extracts of flos Lonicerae, herba Violae, cornu Bubali, Poria, fructus forsythiae, cortex moutan, Coptidis rhizoma, Prunellae Spica and radix astragali, and toothpaste matrix; the traditional Chinese medicine extract comprises the following raw materials in parts by weight: 30-40 parts of honeysuckle, 30 parts of bunge corydalis herb, 15 parts of buffalo horn, 20 parts of red tuckahoe, 15-30 parts of fructus forsythiae, 15 parts of cortex moutan, 3 parts of rhizoma coptidis from Sichuan of China, 15 parts of selfheal and 15-20 parts of astragalus.
2. The compound traditional Chinese medicine toothpaste according to claim 1, wherein the toothpaste base comprises the following components in percentage by weight: 10-25% of silicon dioxide, 35-65% of polyethylene glycol and glycerol, 0.2-1% of carboxymethyl cellulose and xanthan gum, 1.5-2.5% of sodium dodecyl sulfate, 0.1-0.5% of sodium benzoate, 0.1-0.5% of xylitol, 0.5-1.5% of flavoring agent and the balance of water.
3. The compound traditional Chinese medicine toothpaste according to claim 1, wherein the weight percentage of the extract is 0.1-1.5%, and the balance is toothpaste matrix.
4. The compound traditional Chinese medicine toothpaste according to claim 1, wherein the flavoring agent is one of menthol, grapefruit essence, jasmine essence or watermelon essence.
5. The preparation method of the traditional Chinese medicine compound toothpaste with antibacterial, anti-inflammatory and immunity improving functions as claimed in claim 1, which is characterized by comprising the following steps:
(1) adding cornu Bubali into 10-15 times of water, decocting to obtain mixed aqueous solution A;
(2) weighing honeysuckle, corydalis bungeana, poria cocos, fructus forsythiae, cortex moutan, coptis chinensis, selfheal and astragalus membranaceus, mixing the traditional Chinese medicine mixture with 6-8 times of water by mass of the traditional Chinese medicine mixture, adding the traditional Chinese medicine mixture and the water by mass of the traditional Chinese medicine mixture into the mixed aqueous solution A, boiling and extracting, and filtering to obtain liquid medicine;
(3) adding 6-8 times of water into the residue, boiling, extracting, filtering to obtain medicinal liquid, mixing the filtrates, concentrating under reduced pressure to obtain concentrated solution, adding ethanol into the concentrated solution, stirring, standing, filtering to remove precipitate to obtain ethanol precipitation filtrate;
(4) weighing the traditional Chinese medicine combined extract, and uniformly mixing the traditional Chinese medicine combined extract with a toothpaste matrix to obtain the traditional Chinese medicine compound toothpaste.
6. The compound toothpaste of claim 5, wherein in step (4), the toothpaste base comprises silica, polyethylene glycol and glycerin, carboxymethyl cellulose and xanthan gum, sodium dodecyl sulfate, sodium benzoate, xylitol and flavoring agent, and the balance is water.
7. The compound traditional Chinese medicine toothpaste as claimed in claim 5, wherein in step (3), ethanol is added to the concentrated solution to achieve an ethanol content of 40-60% in the concentrated solution.
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| Title |
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| "银花解毒汤", Retrieved from the Internet <URL:https://www.zsbeike.com/yixue/1743528.html> * |
| 罗列娜等: "黄芪及金银花药物牙膏对复发性口腔溃疡的疗效观察", 《新中医》, vol. 46, no. 9, pages 136 - 138 * |
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