CN112826766B - Lucid ganoderma mouthwash and preparation method thereof - Google Patents

Lucid ganoderma mouthwash and preparation method thereof Download PDF

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CN112826766B
CN112826766B CN202110281527.6A CN202110281527A CN112826766B CN 112826766 B CN112826766 B CN 112826766B CN 202110281527 A CN202110281527 A CN 202110281527A CN 112826766 B CN112826766 B CN 112826766B
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ganoderma lucidum
ganoderma
mouthwash
extract
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周岩飞
李晔
林真
陈桂秋
姚渭溪
吴长辉
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Fujian Xianzhilou Biological Science & Technology Co ltd
Ganoherb Bio Technology Fujian Co ltd
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Abstract

The invention belongs to the technical field of daily chemical products, and particularly relates to lucid ganoderma mouthwash and a preparation method thereof. The composite material comprises the following raw materials in parts by weight: 100-200 parts of lucid ganoderma extract, 35-45 parts of erythritol, 0.1-0.2 part of stevioside, 0.4-0.6 part of poloxamer 407, 5-10 parts of fresh flower absolute and 745-860 parts of purified water. The lucid ganoderma mouthwash disclosed by the invention does not contain ethanol or chemical synthetic drugs, is high in safety, can avoid side effects caused by misadministration, and is suitable for long-term use; can inhibit proliferation of harmful bacteria in oral cavity, regulate oral immunity, and assist in repairing recurrent oral ulcer; can avoid tongue coating and tooth staining, has high effective active ingredient concentration, triterpene and sterol concentration not less than 1mg/ml, can maintain pH neutrality in oral environment, and inhibit tooth decalcification and dental plaque formation; fresh breath, cool taste and no residual medicine taste; the preparation method is simple, and the production process is environment-friendly and pollution-free.

Description

Lucid ganoderma mouthwash and preparation method thereof
Technical Field
The invention belongs to the technical field of daily chemical products, and particularly relates to lucid ganoderma mouthwash and a preparation method thereof.
Background
The conventional tooth brushing can only remove 40% -50% of dental plaque and 30% of adjacent dental plaque, and has no effect on removing dental plaque at the edge of gum, between teeth and on the surface of soft tissues of the oral cavity. The mouthwash can deepen all parts of the oral cavity of a person, including the gingival margin and interdental space which are difficult to contact by the toothbrush, and is a good auxiliary plaque control measure. The mouthwash is an indispensable oral cleaning product in modern life, has the advantage of convenient use compared with toothpaste, and is the best choice for guaranteeing the oral hygiene of the old with teeth falling off and the dental patients who cannot tolerate tooth brushing. In recent years, with the increasing health consciousness of people, the requirements for mouthwashes are increasing, and in addition to cleaning teeth, auxiliary prevention of oral diseases is also required.
However, most of traditional mouthwashes are added with alcohol components to play roles in cleaning and sterilizing, but the traditional mouthwashes are strong in irritation and are unacceptable to people. Numerous studies have shown that alcohol has numerous adverse effects that irritate mucous membranes, exacerbate canker sores, cause cancer, poison, and affect the structure of the appliance.
At present, most of alcohol-free mouthwashes use chemical antibacterial agents, such as cetylpyridinium chloride, chlorhexidine, benzalkonium chloride, triclosan and the like, and the chemical antibacterial agents have local side effects and are easy to generate hidden troubles of drug resistance, tooth staining, dry mouth and the like after long-term use. In the prior art, mouthwash schemes of adding natural product extracts are also available, but the antibacterial effect is not ideal due to low content of active ingredients, instability or low antibacterial activity of antibacterial ingredients, and the like, and the stability is poor, and chemical antibacterial agents and chemical preservatives are compounded in most cases.
In recent years, along with the development of the modern technology of natural product extracts, research and utilization methods of active ingredients of the natural product extracts are well developed, and good technical support is provided for mouthwash to utilize the active ingredients of the natural product extracts to assist in preventing oral diseases. However, the natural product extract is mostly brown or black, has bitter and astringent taste and has very heavy traditional Chinese medicine taste, and can keep a certain traditional Chinese medicine taste in the oral cavity after being used as mouthwash, and the color of the extract can dye tongue fur and teeth, so that the extract can cause rejection of users, cannot be popularized and applied, and can only be used as an auxiliary product for oral treatment.
Meanwhile, in terms of safety, some mouthwashes are used without traditional natural products such as antrodia camphorata and the like which are eaten for a long time, and have certain potential toxicity; for example, sweeteners such as sorbitol and xylitol, which do not cause dental caries, are used, but if swallowed excessively by mistake, diarrhea and digestive disorders are caused; the flavoring agent such as menthol or chemical synthetic spice has potential irritation and anaphylaxis.
Disclosure of Invention
The invention aims to overcome the defects of the existing mouthwash, and provides the ganoderma lucidum mouthwash which does not contain alcohol components, does not contain chemical antibacterial agents, is extracted from natural products, has no toxicity, can inhibit proliferation of harmful bacteria in the oral cavity, regulate the oral immune environment, can assist or replace repairing recurrent oral ulcer, can avoid tongue fur and tooth staining, is safe and reliable, and is suitable for long-term use, and the preparation method thereof.
The technical scheme adopted by the invention is as follows: a ganoderma lucidum mouthwash is characterized in that: the composite material comprises the following raw materials in parts by weight: 100-200 parts of lucid ganoderma extract, 35-45 parts of erythritol, 0.1-0.2 part of stevioside, 0.4-0.6 part of poloxamer 407, 5-10 parts of fresh flower absolute and 760-845 parts of purified water.
The invention creatively applies lucid ganoderma to mouthwash, and utilizes the active ingredients of lucid ganoderma to inhibit proliferation of harmful bacteria in the oral cavity and regulate the oral immune environment.
Ganoderma lucidum (Ganoderma lucidum (Leys. Ex Fr.) Karst.) is one of the fungi Ganoderma lucidum of Polyporaceae (part of Chinese pharmacopoeia 2020), has a mild taste, is healthy and healthy, and contains Ganoderma lucidum triterpene and ganoderan as main active ingredients, wherein the Ganoderma lucidum triterpene has various pharmacological effects of regulating immunity, resisting inflammation, resisting allergy, inhibiting bacteria, etc.
With the development of modern natural product separation and detection technology, ganoderma lucidum is used as a safe and effective traditional Chinese medicine with long history, and the efficacy of active ingredients is verified by multiple parties. However, the triterpenes of ganoderma lucidum, which are one of the active ingredients in ganoderma lucidum, have been reported to have remarkable antibacterial effect (Wang Bangwu, etc., "extraction and separation of acid components of ganoderma lucidum, [ J ]", university of Beijing university of technology, 2002, (22): 125-128; wang Xiaoling, etc., "in vitro antibacterial effect of ganoderic acid in ganoderma lucidum mycelia [ J ]", food science and technology, 2008, (10): 184-186). Simultaneously, the ganoderma lucidum also has the effect of regulating the oral immune environment of a human body, and can assist or replace to repair recurrent oral ulcers (Liang Guojian and the like), and clinical observation and discussion of the ganoderma lucidum capsule for treating recurrent afhel ulcers are carried out, chinese medical guidelines, 2011,9 (14): 295-296;Xie L,et al.The effects of freeze-dried Ganoderma lucidum mycelia on a recurrent oral ulceration rat model.BMC Complement Altern Med,2017,17 (1): 511).
The ratio of erythritol and stevioside in the final ganoderma lucidum mouthwash and the proportion of erythritol and stevioside have remarkable influence on the taste masking effect of ganoderma lucidum extract in the ganoderma lucidum mouthwash, and through a large number of experiments and analysis, 35-45 parts of erythritol and 0.1-0.2 part of stevioside have optimal masking effect on the bitter taste of ganoderma lucidum extract and the sensory taste effect of the final ganoderma lucidum mouthwash.
The ratio of the fresh flower absolute to the final lucid ganoderma mouthwash and the ratio of the fresh flower absolute to the poloxamer 407 have remarkable influence on the taste masking effect of the lucid ganoderma extract in the lucid ganoderma mouthwash, and through a large number of experiments and analysis, the masking effect of 5-10 parts of the fresh flower absolute to the traditional Chinese medicine taste of the lucid ganoderma extract and the sensory taste effect of the final lucid ganoderma mouthwash are optimal by 0.4-0.6 part of the poloxamer 407.
Because ganoderma lucidum contains a large amount of melanin, part of melanin and derivatives thereof are soluble in ethanol and are difficult to dissolve in water, and are easy to attach on a solid surface (CN 201910799760.2 is a method for extracting and separating pigment from ganoderma sinensis waste), the conventional ganoderma lucidum extract has the color of dying tongue fur and teeth, and cannot be applied to mouthwash. The invention ensures the effective content of the lucid ganoderma mouthwash by the following inventive preparation method of the lucid ganoderma extract, and overcomes the defect that the lucid ganoderma stains tongue fur and teeth.
The preparation method of the lucid ganoderma extract liquid comprises the following steps:
1. pulverizing dried Ganoderma fruiting body with water content less than or equal to 6%, sieving with 20 mesh sieve to obtain Ganoderma granule, adding 75% ethanol with mass 10 times of Ganoderma granule mass, extracting at 80deg.C for 2 hr, filtering with 200 mesh sieve to obtain filtrate, adding 75% ethanol with mass 10 times of Ganoderma granule mass into the residue again, extracting at 80deg.C for 2 hr, filtering with 200 mesh sieve, mixing the two filtrates, concentrating, oven drying, pulverizing, sieving with 100 mesh sieve to obtain coarse powder of Ganoderma extract;
2. adding 95% ethanol with mass of 8 times of that of coarse powder of Ganoderma extract into coarse powder of Ganoderma extract, dissolving, adding 10% dipotassium hydrogen phosphate aqueous solution with mass of 20 times of that of coarse powder of Ganoderma extract, shake extracting for 10min, standing for 2 hr, and taking out the lower phase to obtain Ganoderma extract.
In the preparation method of the ganoderma lucidum extract, in the step 1, through scientific design of ethanol concentration, feed-liquid ratio, extraction time and extraction times, the content of active ingredients in the obtained ganoderma lucidum extract coarse powder is ensured to be high, the quality is good, so that the final product mouthwash can be ensured to achieve the effects of bacteriostasis, cleaning and adjusting the oral immune environment.
Meanwhile, in the step 2 of the preparation method of the ganoderma lucidum extract, coarse powder of the ganoderma lucidum extract is dissolved through ethanol so as to dissolve melanin and the like in the coarse powder of the ganoderma lucidum extract again, then dipotassium hydrogen phosphate solution is used for double-aqueous phase extraction, the ganoderma lucidum extract obtained by purification is used as a raw material, and water-insoluble melanin components and most of traditional Chinese medicine components in the ganoderma lucidum extract are removed, so that tongue fur and tooth staining can be avoided, and meanwhile, the residual traditional Chinese medicine taste is lighter. Since the aqueous solution of dipotassium hydrogen phosphate is weakly alkaline: the free potassium ions are combined with the ganoderma lucidum triterpene acid which is difficult to dissolve in water to form salt, so that the solubility of the ganoderma lucidum triterpene in ganoderma lucidum extract is greatly improved, and the antibacterial activity of ganoderma lucidum mouthwash is effectively improved; the phosphate is a pH buffer factor naturally existing in the oral cavity of a human body, and the pH of the lucid ganoderma mouthwash is higher than the oral environment of the human body due to the buffer action of the lucid ganoderma extract and the dipotassium hydrogen phosphate supplemented by the final lucid ganoderma mouthwash, so that the acidic environment generated by fermentation food residues of harmful bacteria in the oral cavity is effectively inhibited, the pH neutrality of the oral environment is maintained, and the tooth decalcification and dental plaque formation are inhibited.
The ganoderma lucidum fruiting body is a dried fruiting body of Karst of Polyporaceae fungus ganoderma lucidum Ganoderma lucidum (Leys. Ex Fr.) recorded in the part 2020 of the pharmacopoeia of the people's republic of China, and the triterpene and sterol are calculated by oleanolic acid and are not less than 0.50%.
The method for detecting the triterpenes of the lucid ganoderma in the lucid ganoderma mouthwash is a method for detecting the triterpenes and the sterols under lucid ganoderma items recorded in the part 2020 of pharmacopoeia of the people's republic of China, wherein the concentrations of the triterpenes and the sterols in the lucid ganoderma mouthwash are not less than 1mg/ml calculated by oleanolic acid.
The preparation method of the fresh flower absolute oil provided by the invention comprises the following steps: the preparation method of the medium volatile absolute oil in the method for extracting and separating the components of the red sage flower or the jasmine flower in the patent (CN 201310684930.9) comprises the following raw materials including but not limited to the red sage flower, the jasmine flower or the rose flower, and the like, preferably the red sage flower and the jasmine flower.
Further, the fresh flower absolute according to the invention is a mixture of one or more fresh flower absolute.
The fresh flower absolute is fresh flower absolute of red osmanthus flower or jasmine flower or a mixture of the two fresh flower absolute.
The mixing ratio of the fresh flower absolute oil of the red osmanthus flower or the jasmine flower is 1-3:3-1.
The preparation method of the lucid ganoderma mouthwash comprises the following steps:
1. weighing fresh flower absolute oil and poloxamer 407 according to the formula proportion, and stirring to form uniform oily solution S1;
2. weighing erythritol, stevioside and ganoderma lucidum extract according to the formula proportion, and stirring to form a uniform aqueous solution S2;
3. slowly dripping the S1 into the S2 under the stirring state, uniformly mixing, adding purified water, and uniformly mixing and stirring;
4. stopping stirring, standing at 4deg.C for 24 hr, filtering with 0.22 μm organic film for sterilization, and packaging to obtain Ganoderma gargle.
The steps 3 and 4 of the preparation method of the lucid ganoderma mouthwash have remarkable influence on the sensory taste effect and stability of the lucid ganoderma mouthwash, and through a large number of experiments and analysis, S1 is slowly dripped into S2 under the stirring state, the mixture is uniformly mixed, purified water is added, and the preparation method of uniformly mixing and stirring is optimal for the sensory taste effect and stability of the final lucid ganoderma mouthwash.
Compared with the prior art, the invention has the advantages and beneficial effects that:
1. the lucid ganoderma mouthwash does not contain ethanol or chemical synthetic medicinal components: the ganoderma lucidum triterpene, fresh flower absolute, erythritol and stevioside disclosed by the invention have the advantages of no potential mucosa irritation, canker sore aggravation, cancerogenicity, toxicity, influence on the structure of an appliance, drug resistance, tooth staining or dry mouth and other adverse effects, and the safety is high: lucid ganoderma is currently listed as a test point for both medicine and food, red sage, jasmine flower, rose and the like are traditional edible spice essence raw materials, and erythritol and stevioside are listed in a food additive catalog; the raw materials are not adversely affected by the oral ingestion during daily use, and meanwhile, the raw materials cannot be utilized by microorganisms in the oral cavity and the digestive tract, so that the oral administration preparation is safe and reliable and is suitable for long-term use.
2. The main active ingredient of the ganoderma lucidum mouthwash disclosed by the invention is ganoderma lucidum triterpene, and the antibacterial effect is obvious: has good inhibition effect on streptococcus mutans, lactobacillus acidophilus and porphyromonas gingivalis, has no adverse effects of low content of active ingredients, instability, low antibacterial activity of antibacterial ingredients or unsatisfactory antibacterial effect of other natural products, does not need to additionally add preservative, and is natural, organic and environment-friendly. Simultaneously, the ganoderma lucidum gargle water can effectively reduce local inflammatory reaction and promote tissue repair, and can effectively shorten the healing time of canker sore.
3. The lucid ganoderma mouthwash is prepared by extracting lucid ganoderma extract obtained by extracting and purifying with ethanol and dipotassium hydrogen phosphate solution, and can remove melanin components insoluble in water and most of traditional Chinese medicine components in the lucid ganoderma ethanol extract, so that tongue fur and tooth staining can be avoided, and meanwhile, the residual traditional Chinese medicine taste is lighter. Since the aqueous solution of dipotassium hydrogen phosphate is weakly alkaline: the free potassium ions are combined with the ganoderma lucidum triterpenic acid which is difficult to dissolve in water to form salt, so that the solubility of ganoderma lucidum triterpenoids in ganoderma lucidum extract is greatly improved, the concentration of triterpenoids and sterols in ganoderma lucidum mouthwash is not less than 1mg/ml calculated by oleanolic acid, and the antibacterial activity of ganoderma lucidum mouthwash is effectively improved; the phosphate is a pH buffer factor naturally existing in the oral cavity of a human body, and the pH of the lucid ganoderma mouthwash is higher than the oral environment of the human body due to the buffer action of the lucid ganoderma extract and the dipotassium hydrogen phosphate supplemented by the final lucid ganoderma mouthwash, so that the acidic environment generated by fermentation food residues of harmful bacteria in the oral cavity is effectively inhibited, the pH neutrality of the oral environment is maintained, and the tooth decalcification and dental plaque formation are inhibited.
4. The lucid ganoderma mouthwash disclosed by the invention can obviously inhibit the traditional Chinese medicine taste of lucid ganoderma extract by using fresh flower absolute oil: the fresh flower absolute oil has obvious fragrance, longer duration and fresh breath; the use of erythritol and stevioside can significantly inhibit the bitter taste of the ganoderma lucidum extract: erythritol has cool feel, so that the mouth wash has cool and comfortable taste, does not contain irritants, and has no residual medicine taste in nasal cavities and oral cavities after the mouth wash is spitted out.
5. The lucid ganoderma mouthwash disclosed by the invention is simple in preparation method, environment-friendly in production process and free of pollution.
Drawings
FIG. 1 is a graph showing the experimental results of the effect of Ganoderma mouthwash on the healing time of canker sores
Detailed Description
The present invention will be described in detail with reference to the following examples and drawings to facilitate understanding of the present invention by those skilled in the art. It is specifically pointed out herein that the examples are given solely for the purpose of illustration and are not to be construed as limitations on the scope of the invention, since various modifications and adaptations of the invention will be within the scope of the invention as described above will be apparent to those skilled in the art.
A lucid ganoderma mouthwash comprises the following raw materials in parts by weight: 100-200 parts of lucid ganoderma extract, 35-45 parts of erythritol, 0.1-0.2 part of stevioside, 0.4-0.6 part of poloxamer 407, 5-10 parts of fresh flower absolute and 745-860 parts of purified water. The triterpene and sterol concentration in Ganoderma gargle is not less than 1mg/ml calculated by oleanolic acid. The fresh flower absolute is a mixture of one or more fresh flower absolute, such as the preparation method of the medium volatility absolute described in the method for extracting and separating the components of the red sage flower or the jasmine flower in the patent (CN 201310684930.9), and the raw materials include, but are not limited to, the red sage flower, the jasmine flower or the rose, and the like, preferably the fresh flower absolute of the red sage flower and the jasmine flower, or the mixture of the two fresh flower absolute. The mixing ratio of the fresh flower absolute oil of the red osmanthus flower or the jasmine flower is 1-3:3-1.
The preparation method of the lucid ganoderma extract comprises the following steps: pulverizing dried Ganoderma fruiting body with water content less than or equal to 6%, sieving with 20 mesh sieve to obtain Ganoderma granule, adding 75% ethanol with mass 10 times of Ganoderma granule mass, extracting at 80deg.C for 2 hr, filtering with 200 mesh sieve to obtain filtrate, adding 75% ethanol with mass 10 times of Ganoderma granule mass into the residue again, extracting at 80deg.C for 2 hr, filtering with 200 mesh sieve, mixing the two filtrates, concentrating, oven drying, pulverizing, sieving with 100 mesh sieve to obtain coarse powder of Ganoderma extract; adding 95% ethanol with mass of 8 times of that of coarse powder of Ganoderma extract into coarse powder of Ganoderma extract, dissolving, adding 10% dipotassium hydrogen phosphate aqueous solution with mass of 20 times of that of coarse powder of Ganoderma extract, shake extracting for 10min, standing for 2 hr, and taking out the lower phase to obtain Ganoderma extract.
A preparation method of Ganoderma collutory comprises the following steps: 1. weighing fresh flower absolute oil and poloxamer 407 according to the formula proportion, and stirring to form uniform oily solution S1;
2. weighing erythritol, stevioside and ganoderma lucidum extract according to the formula proportion, and stirring to form a uniform aqueous solution S2;
3. slowly dripping the S1 into the S2 under the stirring state, uniformly mixing, adding purified water, and uniformly mixing and stirring;
4. stopping stirring, standing at 4deg.C for 24 hr, filtering with 0.22 μm organic film for sterilization, and packaging to obtain Ganoderma gargle.
Example 1
A lucid ganoderma mouthwash comprises the following raw materials in parts by weight: 100-200 parts of lucid ganoderma extract, 35-45 parts of erythritol, 0.1-0.2 part of stevioside, 0.4-0.6 part of poloxamer 407, 5-10 parts of fresh flower absolute and 745-860 parts of purified water.
Wherein: the preparation method of the lucid ganoderma extract comprises the following steps:
1. pulverizing dried fruiting body of Ganoderma (moisture less than or equal to 6%), sieving with 20 mesh sieve to obtain 10kg of Ganoderma granule, adding 100kg of 75% ethanol into the Ganoderma granule, extracting at 80deg.C for 2h, filtering with 200 mesh sieve to obtain filtrate, adding 100kg of 75% ethanol into the residue, extracting at 80deg.C for 2h, filtering with 200 mesh sieve, mixing the two filtrates, concentrating, oven drying, pulverizing, sieving with 100 mesh sieve to obtain 705g of Ganoderma extract coarse powder;
2. adding 4kg of 95% ethanol into 500g of ganoderma lucidum extract coarse powder, dissolving, adding 10kg of 10% dipotassium hydrogen phosphate aqueous solution, vibrating and extracting for 10min, standing for 2h, and taking out the phase to obtain about 5kg of ganoderma lucidum extract. A preparation method of Ganoderma collutory comprises the following steps:
1. weighing 100g of pure oil of the fresh osmanthus flowers and 407 g of poloxamer, and stirring to form uniform oily solution S1;2. weighing 450g of erythritol, 2g of stevioside and 2000g of ganoderma lucidum extract, and stirring to form a uniform aqueous solution S2;
3. slowly dripping S1 into S2 at the stirring speed of 300r/min under the stirring state, uniformly mixing, adding about 7450g of purified water, uniformly mixing and stirring at the stirring speed of 300r/min;
4. stopping stirring, standing at 4deg.C for 24 hr, filtering with 0.22 μm organic film, sterilizing, and packaging with 20ml soft aluminum foil bag to obtain Ganoderma gargle.
Example 2
The preparation method of the ganoderma lucidum extract is the same as in example 1.
A preparation method of Ganoderma collutory comprises the following steps:
1. weighing 75g of jasmine flower absolute, and stirring 407 g of poloxamer to form uniform oily solution S1;2. 400g of erythritol and 1.5g of stevioside are weighed, and 1500g of lucid ganoderma extract in example 1 is stirred to form uniform aqueous solution S2;
3. slowly dropwise adding S1 into S2 at the stirring speed of 200r/min under the stirring state, uniformly mixing, adding purified water of about 8000g, uniformly mixing and stirring at the stirring speed of 300r/min;
4. stopping stirring, standing at 4deg.C for 24 hr, filtering with 0.22 μm organic film, sterilizing, and packaging with 20ml soft aluminum foil bag to obtain Ganoderma gargle.
Example 3
The preparation method of the ganoderma lucidum extract is the same as in example 1.
A preparation method of Ganoderma collutory comprises the following steps:
1. weighing flos Osmanthi Fragrantis: jasmine flower is 3:1, 50g of mixed fresh flower absolute, 407 g of poloxamer, and stirring to form uniform oily solution S1;
2. weighing 350g of erythritol, 1.0g of stevioside and 1000g of lucid ganoderma extract in example 1, and stirring to form uniform aqueous solution S2;
3. slowly dripping S1 into S2 at stirring speed of 150r/min under stirring, mixing uniformly, adding 8600g of purified water, mixing uniformly, and stirring at stirring speed of 300r/min;4. stopping stirring, standing at 4deg.C for 24 hr, filtering with 0.22 μm organic film, sterilizing, and packaging with 20ml soft aluminum foil bag to obtain Ganoderma gargle.
The ganoderma lucidum mouthwashes of examples 1, 2 and 3 above were tested:
1. detection of ganoderma triterpene concentration in ganoderma lucidum mouthwash
The ganoderma lucidum mouthwashes of examples 1, 2 and 3 were tested using the method for testing triterpenes and sterols under ganoderma lucidum recorded in the pharmacopoeia 2020 edition of the people's republic of China, the testing results are shown in Table 1
TABLE 1 triterpene and sterol content of Ganoderma mouthwash
2. Antiseptic challenges of ganoderma mouthwash
Examples 1, 2 and 3 were taken as experimental groups and 0.5% of p-hydroxyacetophenone as control group, and mixed bacteria (golden yellow in logarithmic phase) were added respectivelyStaphylococcus liquid, salmonella liquid and escherichia coli liquid are mixed according to the volume ratio of 1:1: 1) and a fungus (Candida albicans) suspension such that the final bacterial content of each sample to be tested is 1 to 5X 10, respectively 6 cfu/g bacteria and 1-5X 10 5 cfu/g fungus. Then, the mixture was thoroughly mixed and incubated at 28 ℃. The total number of bacteria and fungi was measured at 0, 7, 14 and 28 days of inoculation according to the experimental method recommended by the society for cosmetics and fragrance and flavor (CTFA) to determine the preservative efficacy of the ganoderma mouthwash. The judgment standard is as follows: when each sample was inoculated with one inoculation, the viable bacteria amount was reduced to not more than 0.1% of the initial concentration on day 14, followed by gradual reduction and sterile growth on day 28. Then the test is passed; otherwise, the test is not passed.
Table 2 antiseptic challenge test results of Ganoderma mouthwash
From the data in table 2, it can be seen that the ganoderma lucidum mouthwash can pass through the antiseptic challenge excellently, and the antiseptic capability is basically equivalent to that of the pure antiseptic, and the ganoderma lucidum mouthwash has good stability.
3. Antibacterial effect of ganoderma lucidum mouthwash
Experimental grouping:
negative control group: agar plates without liquid medicine; positive control group: the drug concentrations are respectively as follows: 1. 1/2, 1/4, 1/8, 1/16 times the concentration of a brand of mouthwash (ingredients: thymol, eucalyptol, methyl salicylate, menthol) containing a drug; experimental group: the drug concentrations are respectively as follows: 1. 1/2, 1/4, 1/8, 1/16 times the concentration of the ganoderma lucidum mouthwash of examples 1, 2 and 3.
MIC value determination:
the in vitro antibacterial activity of the ganoderma lucidum mouthwash on 3 oral clinical common pathogenic bacteria dominant bacteria is measured by adopting an agar dilution method (Agar dilution method), and is expressed by minimum inhibitory concentration (minimum inhibitory concentration, MIC). Taking experimental bacterial liquid (bacterial concentration: 1.5X10) 8 Per mL), dibbling on the surfaces of agar plates containing drugs with different concentrations, and anaerobic culturing for 48h(80%N 2 +10%CO 2 +10%H 2 ) After that, MIC values are read.
Analysis of results:
in the negative control group (blank group), all experimental bacteria grew well, and MIC values of the positive control group and the experimental group drug aqueous solution could be included in the result analysis, as shown in table 3.
TABLE 3 MIC values of Ganoderma mouthwash for 3 oral bacteria (times)
As can be seen from Table 3, the MIC values of the ganoderma lucidum mouthwash for 3 oral bacteria are smaller than those of the positive control group, namely, the ganoderma lucidum mouthwash has stronger inhibition effect on streptococcus mutans, lactobacillus acidophilus and porphyromonas gingivalis than common mouthwashes on the market. The minimum MIC value of the lucid ganoderma mouthwash to 3 oral clinical common pathogenic bacteria dominant bacteria can reach 1/8 concentration, namely the lucid ganoderma mouthwash still has a certain inhibition effect on streptococcus mutans and lactobacillus acidophilus after being diluted to 8 times, and still has a certain inhibition effect on porphyromonas gingivalis after being diluted to 4 times.
4, the ganoderma lucidum mouthwash has the treatment effect on rat canker sore
Establishing an oral ulcer model:
rats were anesthetized with 2% sodium pentobarbital, naOH crystals were cauterized at the mucous membrane of the cheek side of the two sides of the rat for 60s, then washed with physiological saline, and observed after 30 min:
experimental grouping and treatment procedure:
52 rats were randomly assigned to the ganoderma lucidum mouthwash experimental group (examples 1, 2 and 3), the control group and the blank group (no ulcers), 10 each, and the remaining 2 were model groups. Observations were made at 24, 48, 72 and 96 hours and at the final healing, respectively, after 24h modeling was successful, during which oral mucosal application was performed daily. The experimental groups are respectively adsorbed with cotton swabs to corresponding 0.5mL of liquid medicine, and the control group is applied with 0.5mL of 0.9% sodium chloride solution for 5min at the oral ulcer of the rat. Blank and model groups were not treated. Each group of rats was obtained by taking 2 rats at 72 hours, the remaining rats were sacrificed when the ulcers were substantially healed, and mucous membranes at the dental ulcers were taken, and the healing time of each group was recorded.
Analysis of results:
the oral ulcer healing time of examples 1, 2 and 3 was significantly shorter than that of the control group (P < 0.01); the difference in the time to healing of the canker sore in the groups of examples 1 and 2 was not statistically significant (P > 0.05). See fig. 1.
As can be seen from fig. 1, the ganoderma lucidum mouthwash has a remarkable shortening effect on the healing time of dental ulcers, and the ganoderma lucidum mouthwash has an effect on the tissue repair and change of dental ulcers of rats:
hematoxylin-eosin staining (HE staining) results showed: after 72 hours of administration, the superficial epithelium and lamina propria necrosis of the focal zone of examples 1 and 2 show obvious tissue edema, and a great deal of inflammatory cells such as phagocytes, lymphocytes and the like infiltrate, and the newly-born granulation tissues enrich blood vessels and form; example 3 group lamina propria is essentially necrotic and inflammatory cells are extensively infiltrated; control group ulcer area epithelium and lamina propria necrosis, inflammatory cell infiltration, granulation tissue formation, capillary formation, myoblast degeneration and necrosis; the oral mucosa of the rats in the blank group is normal mucosa without ulcer, the epithelium and the lamina propria are continuous and have a certain thickness, and tissue necrosis, cell edema, inflammatory cell infiltration and the like are avoided; the ulcerated area of the model group was seen as partial epithelial lysis, continuous disappearance, edema of connective tissue layer, and massive infiltration of neutrophils, monocytes and macrophages. The granulation tissue of the groups 1, 2 and 3 grows well after 72 hours of administration, and the healing degree is not obviously different under a light microscope; the control group has obvious ulcer local inflammatory symptoms and poor healing.
According to the ganoderma lucidum mouthwash formula, 100-200 parts of ganoderma lucidum extract, 35-45 parts of erythritol, 0.1-0.2 part of stevioside, 407.4-0.6 part of poloxamer, 5-10 parts of fresh flower absolute and 745-860 parts of purified water are obtained through a large number of experiments and analysis, and all raw materials in the formula jointly play roles, so that the ganoderma lucidum mouthwash formula has a good inhibition effect on streptococcus mutans, lactobacillus acidophilus and porphyromonas gingivalis, can effectively reduce local inflammatory reaction, promote tissue repair and effectively shorten the healing time of canker sores.

Claims (6)

1. The lucid ganoderma mouthwash is characterized by comprising the following raw materials in parts by weight: 100-200 parts of lucid ganoderma extract, 35-45 parts of erythritol, 0.1-0.2 part of stevioside, 0.4-0.6 part of poloxamer 407, 5-10 parts of fresh flower absolute and 745-860 parts of purified water, wherein the preparation method of the lucid ganoderma extract comprises the following steps:
1.1, crushing dried ganoderma lucidum fruiting bodies with water content less than or equal to 6%, sieving with a 20-mesh sieve to obtain ganoderma lucidum particles, adding 75% ethanol with the mass 10 times of the mass of the ganoderma lucidum particles into the ganoderma lucidum particles, extracting at 80 ℃ for 2h, filtering with a 200-mesh filter screen to obtain filtrate, adding 75% ethanol with the mass 10 times of the mass of the ganoderma lucidum particles into filter residues again, extracting at 80 ℃ for 2h, filtering with a 200-mesh filter screen, combining the two extracted filtrate, concentrating, drying, crushing, sieving with a 100-mesh sieve to obtain ganoderma lucidum extract coarse powder;
1.2 adding 95% ethanol with the mass of 8 times of the mass of the coarse powder of the ganoderma lucidum extract into the coarse powder of the ganoderma lucidum extract, dissolving, adding 10% dipotassium hydrogen phosphate aqueous solution with the mass of 20 times of the mass of the coarse powder of the ganoderma lucidum extract, carrying out shake extraction for 10min, standing for 2h, and taking out the phase to obtain the ganoderma lucidum extract.
2. The ganoderma lucidum mouthwash according to claim 1, wherein the concentration of triterpenes and sterols in the ganoderma lucidum mouthwash is not less than 1mg/ml in terms of oleanolic acid.
3. The ganoderma lucidum mouthwash of claim 1, wherein the fresh flower absolute is a mixture of one or more fresh flower absolute.
4. A ganoderma lucidum mouth wash according to claim 3 wherein the fresh flower absolute is that of red sage or jasmine or a mixture of both fresh flower absolute.
5. The ganoderma lucidum mouthwash according to claim 4, wherein the mixing ratio of fresh flower absolute of red sage or jasmine is 1-3:3-1.
6. A method of preparing the ganoderma lucidum mouthwash of claim 1, characterized in that:
6.1. weighing fresh flower absolute oil and poloxamer 407 according to the formula proportion, and stirring to form uniform oily solution S1;
6.2. weighing erythritol, stevioside and ganoderma lucidum extract according to the formula proportion, and stirring to form a uniform aqueous solution S2;
6.3. slowly dripping the S1 into the S2 under the stirring state, uniformly mixing, adding purified water, and uniformly mixing and stirring;
6.4. stopping stirring, standing at 4deg.C for 24 hr, filtering with 0.22 μm organic film for sterilization, and packaging to obtain Ganoderma gargle.
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CN103695175A (en) * 2013-12-13 2014-04-02 仙芝科技(福建)股份有限公司 Method for extracting and separating components of osmanthus flower or arabian jasmine flower
CN104188889A (en) * 2014-08-27 2014-12-10 仙芝科技(福建)股份有限公司 Lucid ganoderma toothpaste
CN104490742A (en) * 2014-11-25 2015-04-08 张家港保税区冠祥贸易有限公司 Cavity health care cleaning fluid and producing method thereof
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CN103695175A (en) * 2013-12-13 2014-04-02 仙芝科技(福建)股份有限公司 Method for extracting and separating components of osmanthus flower or arabian jasmine flower
CN104188889A (en) * 2014-08-27 2014-12-10 仙芝科技(福建)股份有限公司 Lucid ganoderma toothpaste
CN104490742A (en) * 2014-11-25 2015-04-08 张家港保税区冠祥贸易有限公司 Cavity health care cleaning fluid and producing method thereof
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