Preparation method of compound traditional Chinese medicine spray for treating oral cavity infection

Abstract

The present application relates to a kind of preparation method of compound traditional Chinese medicine spray for treating oral infection, the compound traditional Chinese medicine spray provided in the present application selects traditional Chinese medicine such as Chinese herbaceous peony, honeysuckle, baizhi, mint, material is widely selected, and nontoxic and harmless, honeysuckle and Chinese herbaceous peony are main medicine, baizhi is adjuvant, mint improves taste, the present application also explores the inhibitory effect of the best proportion of traditional Chinese medicine on staphylococcus aureus, streptococcus pneumoniae and other common pathogenic bacteria of respiratory tract, the compound traditional Chinese medicine oral spray provided in the present application is used, and the effect of oral infection caused by staphylococcus aureus, streptococcus pneumoniae and the like is obvious.

Description

Technical Field

[0001] This invention belongs to the field of compound traditional Chinese medicine for treating oral infections. Specifically, it is a compound traditional Chinese medicine spray for treating oral infections. A method for preparing a compound traditional Chinese medicine spray for treating oral infections, composed of Lithospermum erythrorhizon, Angelica dahurica, Lonicera japonica, and Mentha haplocalyx, is disclosed. Background Technology

[0002] Recent studies have shown that oral and respiratory tract infections are common clinical conditions with a high recurrence rate, and they easily spread to adjacent areas. Common pathogens include Streptococcus pneumoniae, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa. In-depth research into the drug resistance of these pathogens has revealed that resistance is steadily increasing, posing challenges to clinical treatment. There is an urgent need in this field for a traditional Chinese medicine spray that can be used to treat oral infections caused by Staphylococcus aureus, Streptococcus pneumoniae, and other bacteria. Summary of the Invention

[0003] This invention provides a method for preparing a compound traditional Chinese medicine spray to treat oral infections caused by Staphylococcus aureus, Streptococcus pneumoniae, etc., thereby improving the therapeutic effect. To achieve the above objective, this invention discloses a method for preparing a compound traditional Chinese medicine spray for treating oral infections, composed of Lithospermum erythrorhizon, Lonicera japonica, Angelica dahurica, and Mentha haplocalyx. The specific steps are as follows:

[0004] (1) Extraction of drugs: Accurately weigh Angelica dahurica, Lithospermum erythrorhizon and Lonicera japonica, add 200mL of 15-95% ethanol to soak for 1-3 days, filter out the drug solution, evaporate each group of filtrate using a rotary evaporator, add distilled water to the extract to make up to a final drug concentration of 500mg / ml, and refrigerate for later use.

[0005] (2) Dissolve peppermint crystals in anhydrous ethanol to prepare a peppermint solution of 0.5 g / ml. Refrigerate the extracted drug for later use.

[0006] (3) Using 20% ​​ethanol as the substrate, the honeysuckle, purple gromwell, angelica and peppermint were mixed in a mass-volume ratio of 4:4:2:1 to prepare a compound Chinese medicine oral spray. The volume ratio of 20% ethanol to the mixed medicine was 1:4 to 1:8.

[0007] Furthermore, in step (3), xylitol of 1% by total volume can be added to the mixed drugs to improve the taste.

[0008] In a preferred embodiment of the present invention, in step (1), Angelica dahurica, Lithospermum erythrorhizon and Lonicera japonica are extracted by soaking in 50% ethanol.

[0009] In a preferred embodiment of the present invention, in step (3), the volume ratio of 20% ethanol to the mixed drug is 1:6.

[0010] Many components of traditional Chinese medicine, such as alkaloids, organic acids, saponins, and volatile oils, have certain antibacterial effects. Moreover, traditional Chinese medicine has advantages such as abundant resources, wide availability, and low likelihood of developing drug resistance, and is widely used in the treatment of oral diseases.

[0011] Lithospermum has a long history of use in my country and is widely used clinically. It is cold in nature and has the effects of clearing heat and cooling blood, promoting blood circulation and detoxifying, and promoting rash eruption and fading spots. Lithospermum mainly contains naphthoquinones, terpenes, steroids, phenolic acids, alkaloids, and polysaccharides, which have anti-inflammatory, anti-tumor, antiviral, liver-protective, anti-fertility, and burn and wound-healing effects.

[0012] Honeysuckle is the dried flower bud or newly opened flower of *Lonicera japonica*, a plant in the Caprifoliaceae family. It contains volatile oils, organic acids, flavonoids, triterpenes, and saponins, among other active substances, and possesses antibacterial, antioxidant, antipyretic, anti-inflammatory, antiviral, immunomodulatory, and hepatoprotective and choleretic effects. In recent years, the anti-inflammatory pharmacological activity of honeysuckle, a traditional Chinese medicine for clearing heat and detoxifying, has increasingly become a research hotspot for scholars both domestically and internationally.

[0013] Angelica dahurica is a commonly used traditional Chinese medicine. It mainly contains chemical components such as coumarins, volatile oils, polysaccharides, alkaloids, and amino acids, and has pharmacological effects such as analgesia, anti-inflammation, anti-oxidation, anti-tumor, and whitening.

[0014] Peppermint is the dried aerial part of the plant *Mentha haplocalyx*, belonging to the Lamiaceae family. It has a pungent taste and cool properties, and enters the lung and liver meridians. It possesses the effects of dispersing wind-heat, clearing the head and eyes, relieving sore throat and promoting rash eruption, and soothing the liver and regulating qi. As a commonly used pungent and cool exterior-releasing herb, peppermint's main components include volatile oils, flavonoids, terpenes, phenolic acids, quinones, and phenylpropanoids. Modern pharmacological studies have shown that it has antibacterial, antiviral, anti-inflammatory, antioxidant, antitumor, and antifertility effects.

[0015] The compound traditional Chinese medicine spray provided by this invention selects herbs such as Lithospermum erythrorhizon, honeysuckle, Angelica dahurica, and peppermint. The materials are widely available, non-toxic, and harmless. Honeysuckle and Lithospermum erythrorhizon are the main ingredients, Angelica dahurica is the auxiliary ingredient, and peppermint improves the taste. This invention also explores the inhibitory effect of the optimal ratio of traditional Chinese medicine on common respiratory pathogens such as Staphylococcus aureus and Streptococcus pneumoniae. The compound traditional Chinese medicine oral spray provided by this invention has a significant effect on treating oral infections caused by Staphylococcus aureus, Streptococcus pneumoniae, etc. Attached Figure Description

[0016] Figure 1 Antibacterial effects of different extracts on Streptococcus pneumoniae;

[0017] Figure 2 Antibacterial effects of different extracts on Staphylococcus aureus;

[0018] Figure 3 Antibacterial effects of different drug ratios. Detailed Implementation

[0019] The present invention is described in detail below through specific embodiments, but this does not limit the scope of protection of the present invention. Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and the experimental equipment, materials, reagents, etc. used can all be obtained commercially.

[0020] Example 1

[0021] (1) Extraction of the drug: Accurately weigh six 25g portions of honeysuckle, add 200ml of 15-95% ethanol (five points of 95%, 75%, 50%, 25%, and 15% were taken for comparison) and water (drug:water = 1:8) to each portion, soak for 2 days, filter out the liquid, and evaporate the filtrate of each group using a rotary evaporator (EYEL4N-1300 Tokyo RIKEN Equipment Co., Ltd.). The extract is diluted with distilled water to a final drug concentration of 500mg / ml. The above-mentioned extracted drugs are refrigerated for later use.

[0022] (2) The extraction methods for Angelica dahurica and Lithospermum erythrorhizon are the same as those for Lonicera japonica.

[0023] (3) Dissolve 1g of peppermint crystals in 2ml of anhydrous ethanol to prepare a peppermint solution with a concentration of 0.5g / ml. Refrigerate the extracted drug for later use.

[0024] (4) Preparation of drug sensitivity test discs: 20 μL of extracted drug is added to a sterile blank drug sensitivity test disc to prepare a drug sensitivity test disc.

[0025] (5) Prepare a Staphylococcus aureus and Streptococcus pneumoniae suspension with a turbidity of 0.5 McFarland and serially dilute it to 10 with physiological saline. 6 cfu / ml.

[0026] The antibacterial effects of Lithospermum erythrorhizon, Lonicera japonica, and Angelica dahurica were determined using the disk diffusion method (KB method). 50 μL of bacterial suspension was evenly spread onto pre-prepared nutrient agar plates, and prepared antimicrobial susceptibility testing disks were added. After incubation at 37℃ for 20 hours, measurements were taken three times using vernier calipers, and the average value was recorded. The diameter of the inhibition zone indicated the strength of the antibacterial effect. When the dosage and incubation time were the same, Lithospermum erythrorhizon, Angelica dahurica, and Lonicera japonica all showed certain antibacterial effects against Streptococcus pneumoniae and Staphylococcus aureus, with the drugs extracted using 50% ethanol showing the best antibacterial effect. Results are shown below. Figure 1 , Figure 2 .

[0027] The antibacterial effects of Lithospermum erythrorhizon, Lonicera japonica, and Angelica dahurica were determined using a suspension quantitative method. The KB method showed that the drugs extracted with 50% ethanol had the best antibacterial effect. Therefore, Lithospermum erythrorhizon, Lonicera japonica, and Angelica dahurica extracted with 50% ethanol were selected and serially diluted with sterile physiological saline (NS) to different concentrations of 3:1, 2:1, 1:1, 1:2, 1:4, 1:6, and 1:8, respectively, for single-drug antibacterial experiments. 50 μL of bacterial suspension (concentration of 10) was used. 6 Add 450 μL of the drug (cfu / mL) to a plate, using sterile normal saline (NS) as a control. After mixing, pour 50 μL of the solution onto a plate and incubate at 37°C for 24 h. Count the colonies. Repeat the experiment twice, take the average value, and calculate the result.

[0028] Viable bacteria counting method (cfu / mL) = plate count result × 10.

[0029] Sterilization rate = (logarithm of viable bacteria count before sterilization - logarithm of viable bacteria count after sterilization) / logarithm of viable bacteria count before sterilization × 100%

[0030] Using a suspension quantitative method, it was found that Lithospermum erythrorhizon, Angelica dahurica, and Lonicera japonica all had significant antibacterial effects against Streptococcus pneumoniae and Staphylococcus aureus. The results are shown in Table 1.

[0031] Table 1. Antibacterial effects of single drugs at different concentrations

[0032]

[0033] Example 2: Antibacterial effect of mixed drugs

[0034] The KB method results showed that honeysuckle and lithospermum had strong antibacterial effects, so these two herbs were selected as the main drugs, angelica dahurica as the auxiliary drug, and peppermint to improve the taste, for a mixed drug antibacterial experiment. Eight mixed drugs were prepared with mass-volume ratios of honeysuckle:lithospermum:angelica dahurica:pemint of 4:4:2:1, 3:5:2:1, 2:6:2:1, 5:3:2:1, 6:2:2:1, 4:4:2.5:0.5, 4:4:1.5:1.5, and 4:3:3:1, respectively. The test bacteria were Staphylococcus aureus and Staphylococcus epidermidis. 20 μL of the drug solution and 50 μL of bacterial suspension (concentration 10) were taken. 5 Paper disc diffusion experiments were conducted using cfu / ml. Results are shown below. Figure 3 The experimental results show that the optimal mass-volume ratio of the ingredients in this compound Chinese herbal oral spray—honeysuckle, purple gromwell, angelica, and peppermint—is 4:4:2:1, which yields the best therapeutic effect for oral infections caused by Staphylococcus aureus, Streptococcus pneumoniae, and other bacteria.

[0035] Example 3: Antibacterial effect of drug solutions diluted at different ratios

[0036] The antibacterial experiment of different concentrations of the mixed drug was conducted by serially diluting the mixed drug to different concentrations of 3:1, 2:1, 1:1, 1:2, 1:4, 1:6, 1:8, and 1:12. Physiological saline and 20% ethanol were used as controls. The suspension quantitative method was performed using the same method as described above. The test bacteria were Staphylococcus aureus and Streptococcus pneumoniae. The results are shown in Table 2.

[0037] Table 2. Antibacterial effect of drug solutions diluted at different ratios

[0038]

[0039] The bactericidal rate of the diluted medicinal solution reached over 99% at different ratios, and 20% ethanol also had a bactericidal effect. Using 20% ​​ethanol as the substrate, a compound traditional Chinese medicine oral spray was prepared by mixing honeysuckle, purple gromwell, angelica dahurica, and peppermint in a mass-to-volume ratio of 4:4:2:1. Soaking the mixed medicinal materials in 20% ethanol and diluting the extract at a volume ratio of 1:6 yielded better results.

[0040] Traditional Chinese medicine has a bitter taste, so xylitol and peppermint were selected to improve its flavor. Xylitol (dosage: 1% of the total volume of the compound traditional Chinese medicine spray) and 0.5 mg / ml peppermint solution were used as flavoring agents. See Table 3.

[0041] Table 3. Effects of different ingredients on taste

[0042]

[0043] The experimental results show that adding 1% xylitol and 0.5mg / ml of peppermint to the total volume of the compound Chinese medicine spray results in the best taste, with a slightly sweet and refreshing flavor.

[0044] In summary, the optimal mass-volume ratio of the components honeysuckle, purple gromwell, angelica dahurica, and peppermint in the compound traditional Chinese medicine oral spray provided by this invention is 4:4:2:1, which has a significant effect on treating oral infections caused by Staphylococcus aureus, Streptococcus pneumoniae, etc.

[0045] Example 4

[0046] (1) Extraction of drugs: accurately weigh Angelica dahurica, Lithospermum erythrorhizon and Lonicera japonica, add 200 mL of 50% ethanol to soak for 2 days, filter out the drug solution, evaporate each group of filtrate using a rotary evaporator, add distilled water to the extract to make up to a final drug concentration of 500 mg / ml, and refrigerate for later use.

[0047] (2) Dissolve 1g of peppermint crystals in 2mL of anhydrous ethanol to prepare a peppermint solution of 0.5g / ml. Refrigerate the extracted drug for later use.

[0048] (3) Using 20% ​​ethanol as the substrate, a compound traditional Chinese medicine oral spray was prepared by mixing honeysuckle, purple gromwell, angelica dahurica, and peppermint in a mass-to-volume ratio of 4:4:2:1. The volume ratio of 20% ethanol to the mixed drugs was 1:6, and xylitol was added at 1% of the total volume of the mixed drugs. The preparation of the spray can be carried out by any method known in the art, such as sterilizing the well-stirred mixed drugs and then filling them into an oral sprayer with a suction nozzle.

[0049] The above description is merely a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for preparing a compound traditional Chinese medicine spray for treating oral infections, characterized in that, Includes the following steps: (1) Extraction of drugs: accurately weigh Angelica dahurica, Lithospermum erythrorhizon and Lonicera japonica, add 200 mL of 50% ethanol to soak for 1 to 3 days, filter out the drug solution, evaporate the filtrate of each group using a rotary evaporator, add distilled water to the extract to make up to a final drug concentration of 500 mg / ml, and refrigerate for later use. (2) Dissolve peppermint crystals in anhydrous ethanol to prepare peppermint solution, extract the drug and refrigerate for later use; (3) Prepare a compound Chinese medicine oral spray by mixing honeysuckle, purple gromwell, angelica and peppermint in a mass-volume ratio of 4:4:2:

1. Select 20% ethanol as substrate and add the mixed drugs to the substrate. The volume ratio of 20% ethanol to the mixed drugs is 1:4 to 1:

8.

2. The preparation method according to claim 1, characterized in that, In step (2), 1g of peppermint crystals are dissolved in 2mL of anhydrous ethanol to prepare a peppermint solution of 0.5g / ml.

3. The preparation method according to claim 1, characterized in that, In step (3), the volume ratio of 20% ethanol to the mixed drugs is 1:

6.

4. The preparation method according to claim 1, characterized in that, In step (3), xylitol of 1% of total volume may also be added to the mixed drugs.

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