Preparation method of chamomilla recutita flower extract and oral care composition containing chamomilla recutita flower extract

Abstract

The invention relates to the technical field of oral care ingredients, and discloses a preparation method of a chamomilla recutita flower extract and an oral care composition containing the chamomilla recutita flower extract, the chamomilla recutita flower extract is obtained by soaking chamomilla recutita flower growing in mediterranean climate in 1, 2-propylene glycol and water, draining water, pressurizing and filtering; the oral care composition comprises the following components: a chamomilla recutita flower extract, sodium monofluorophosphate and water. The chamomilla recutita flower extract capable of effectively inhibiting expression of inflammatory factor prostaglandin E2 (PGE2) and inhibiting growth of porphyromonas gingivalis and streptococcus mutans is screened and compounded with sodium monofluorophosphate to form the oral care composition, and the oral care composition has good effects of inhibiting bacteria, relieving gingivitis and relieving halitosis.

Description

Technical Field

[0001] This invention relates to the field of oral care ingredients, and in particular to a method for preparing chamomile extract and an oral care composition containing therein. Background Technology

[0002] Oral diseases are among the most common health problems, including tooth decay, gingivitis, and halitosis. These diseases can lead to impaired oral function, pain, and tooth damage, thus affecting both physical and mental health. Therefore, oral health is receiving increasing attention.

[0003] Dental caries is caused by the demineralization of the hard tissues of the tooth (enamel and dentin) due to organic acids produced during bacterial fermentation. The endogenous bacteria in the biofilm are mainly *Streptococcus mutans* and *Lactobacillus*. These bacteria produce weak organic acids that lower the local pH below a critical value, leading to demineralization of the tooth tissue. *Streptococcus mutans* is a Gram-positive bacterium and the most abundant species of *Streptococcus* in the natural oral flora. It is a major component of dental plaque and one of the main bacteria causing dental caries.

[0004] Gingivitis is a mild, reversible periodontal disease that can develop into periodontitis if left untreated. The main approach to treating gingivitis is to control plaque buildup and soft tissue inflammation. Current guidelines consider chlorhexidine the gold standard for preventing and treating gingivitis; however, chlorhexidine has many negative side effects, such as erosion of the oral mucosa, tooth discoloration, and a bitter taste.

[0005] Halitosis is caused by a large number of microorganisms in the oral cavity. These microorganisms decompose and metabolize food debris such as amino acids and proteins, producing unpleasant odors—volatile sulfur compounds (VSCs). The main components of these sulfides are hydrogen sulfide, methyl mercaptan, and ethyl sulfides. Clinical trials have shown a positive correlation between VSC levels and the severity of halitosis. *Porphyromonas gingivalis*, a Gram-negative bacterium, decomposes amino acids and proteins to produce hydrogen sulfide (H₂S) and methyl mercaptan (CH₃SH), and is closely related to halitosis.

[0006] Matricaria is a type of herbaceous plant belonging to the Asteraceae family, possessing anti-inflammatory, antibacterial, antioxidant, wound-healing-promoting, and analgesic properties. Existing technologies include research on the application of chamomile extract in oral care products. For example, patent CN116710065A discloses an oral composition containing plant extracts, preferably at least one selected from the following: Alpinia oxyphylla extract, Paeonia lactiflora extract, Angelica sinensis extract, Calendula officinalis extract, Matricaria champaca extract, and Sapindus mukorossi extract.

[0007] However, the types and contents of active ingredients in chamomile plant extracts are closely related to the origin of chamomile, the part from which it is extracted, and the extraction method. Existing chamomile extracts have limited effects in inhibiting the expression of inflammatory factors and the growth of Porphyromonas gingivalis and Streptococcus mutans, and their antibacterial, gingivitis-relieving, and halitosis-reducing effects are not good. Summary of the Invention

[0008] The present invention aims to overcome the aforementioned problems of existing chamomile extracts and provides a method for preparing chamomile extract and an oral care composition containing it. The method screens out chamomile extracts that can effectively inhibit the expression of the inflammatory factor prostaglandin E2 (PGE2) and inhibit the growth of Porphyromonas gingivalis and Streptococcus mutans. When combined with sodium monofluorophosphate, this chamomile extract is used as an oral care composition, exhibiting good antibacterial, gingivitis-relieving, and halitosis-reducing effects.

[0009] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a method for preparing chamomile extract, comprising the following steps: (1) After drying the chamomile flowers grown in the Mediterranean climate, sieve them to obtain chamomile powder. Soak the chamomile powder in a mixed solvent of 1,2-propanediol and water. The volume ratio of 1,2-propanediol to water in the mixed solvent is 0.5~1.5:1, and the soaking time is 2~4 days. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract.

[0010] This invention selects chamomile flowers grown in the Mediterranean climate, characterized by hot, dry summers and mild, rainy winters, for extraction. Chamomile flowers grown in the mild, humid Mediterranean climate with suitable sunlight have a higher content of active ingredients such as flavonoids. The extraction process uses a mixed solvent of 1,2-propanediol and water. Water, being a polar solvent, is suitable for extracting water-soluble components, such as flavonoids, but it is prone to microbial growth and requires the addition of preservatives. 1,2-propanediol has preservative properties and a higher extraction rate for fat-soluble components, such as bisabolol, but its ability to extract water-soluble substances is weaker. This invention uses 1,2-propanediol and water in a volume ratio of 0.5~1.5:1 as a mixed solvent, which can accommodate both water-soluble and lipid-soluble active ingredients. Furthermore, 1,2-propanediol can improve solution permeability, thereby more effectively extracting the active ingredients from chamomile cells (common alcohols such as 1,3-propanediol, 1,3-butanediol, and glycerol have weaker preservative effects and permeability than 1,2-propanediol). The chamomile extract obtained by this invention can effectively inhibit the expression of the inflammatory factor prostaglandin E2 (PGE2) and inhibit the growth of Porphyromonas gingivalis and Streptococcus mutans. When used as an ingredient in oral care products, it can have good antibacterial, gingivitis-relieving, and halitosis-reducing effects.

[0011] As a preferred option, in step (1), the chamomile flowers grown in the Mediterranean climate are dried and passed through an 80-100 mesh sieve.

[0012] Preferably, the mass-to-volume ratio of chrysanthemum powder to mixed solvent during soaking in step (1) is 1g:5~10mL.

[0013] Preferably, in step (1), the soaking is carried out at room temperature and stirred.

[0014] Preferably, the pressure during pressurization in step (2) is 0.3~0.4MPa and the pressurization time is 0.5~2h.

[0015] Secondly, the present invention provides a chamomile extract prepared by the above method.

[0016] Preferably, the total phenol content is ≥0.07% and the total flavonoid content is ≥0.06% by mass fraction.

[0017] As a preferred option, the content of apigenin 7-glucoside, a flavonoid, is ≥0.01% by mass fraction.

[0018] Thirdly, the present invention provides an oral care composition comprising the above-mentioned chamomile extract, sodium monofluorophosphate, and water.

[0019] Sodium monofluorophosphate is an inorganic compound primarily used as an anti-caries agent and tooth desensitizer. In aqueous solution, sodium monofluorophosphate releases fluoride ions, which penetrate tooth enamel and combine with tooth minerals (such as hydroxyapatite) to form a stronger fluorapatite, effectively preventing tooth demineralization and enhancing the tooth's resistance to acid erosion. This invention uses sodium monofluorophosphate and chamomile extract as effective components in an oral care composition, exhibiting synergistic antibacterial and anti-inflammatory effects, enhancing the composition's antibacterial, gingivitis-soothing, and halitosis-reducing effects.

[0020] Preferably, the components, by mass percentage, include 0.5-1.5% sodium monofluorophosphate, 0.0125-0.1% chamomile extract, and the balance being water.

[0021] Therefore, the present invention has the following beneficial effects: (1) A magnolia extract grown in the Mediterranean climate was prepared by extracting it with a mixed solvent of 1,2-propanediol and water. The extract could effectively inhibit the expression of the inflammatory factor prostaglandin E2 (PGE2) and inhibit the growth of Porphyromonas gingivalis and Streptococcus mutans. (2) Sodium monofluorophosphate and chamomile extract are used together as effective components of oral care composition. They have synergistic antibacterial and anti-inflammatory effects, which can enhance the antibacterial, gingivitis-relieving and halitosis-reducing effects of the composition. Detailed Implementation

[0022] The present invention will be further described below with reference to specific embodiments.

[0023] In this invention, unless otherwise specified, all equipment and raw materials are available from the market or commonly used in the industry. Unless otherwise specified, the methods in the following embodiments are conventional methods in the art.

[0024] Example 1: An oral care composition comprising, by weight percentage: 0.5% sodium monofluorophosphate, 0.0125% chamomile extract and 99.4875% water.

[0025] The preparation method of chamomile extract is as follows: (1) After drying the chamomile grown in the Mediterranean climate region of France, pass it through a 100-mesh sieve to obtain chamomile powder; take 10g of chamomile powder and soak it in a mixed solvent of 90mL of 1,2-propanediol and water at room temperature for 3 days with stirring; the volume ratio of 1,2-propanediol and water in the mixed solvent is 1:1. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract; the pressure during pressurization is 0.35 MPa and the pressurization time is 1 hour; the chamomile extract obtained has a total phenol content of 0.075 wt%, a total flavonoid content of 0.067 wt%, and a flavonoid content of apigenin 7-glucoside of 0.014 wt%.

[0026] Example 2: An oral care composition comprising, by weight percentage: 0.98% sodium monofluorophosphate, 0.0125% chamomile extract and 99.0075% water.

[0027] The preparation method of chamomile extract is as follows: (1) After drying the chamomile flowers grown in the Mediterranean climate region of France, pass them through a 100-mesh sieve to obtain chamomile powder; take 10g of chamomile powder and soak it in a mixed solvent of 90mL of 1,2-propanediol and water at room temperature for 3 days with stirring; the volume ratio of 1,2-propanediol and water in the mixed solvent is 1:1. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract; the pressure during pressurization is 0.35 MPa and the pressurization time is 1 hour.

[0028] Example 3: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0029] The preparation method of chamomile extract is as follows: (1) After drying the chamomile flowers grown in the Mediterranean climate region of France, pass them through a 100-mesh sieve to obtain chamomile powder; take 10g of chamomile powder and soak it in a mixed solvent of 90mL of 1,2-propanediol and water at room temperature for 3 days with stirring; the volume ratio of 1,2-propanediol and water in the mixed solvent is 1:1. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract; the pressure during pressurization is 0.35 MPa and the pressurization time is 1 hour.

[0030] Comparative Example 1: The sample in Comparative Example 1 contained only water and no sodium monofluorophosphate or chamomile extract was added.

[0031] Comparative Example 2: An oral care composition comprising, by weight percentage: 0.5% sodium monofluorophosphate and 99.5% water.

[0032] Comparative Example 3: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate and 98.5% water.

[0033] Comparative Example 4: An oral care composition comprising, by weight percentage: 0.0125% chamomile extract and 99.9875% water; the chamomile extract is prepared in the same manner as in Example 1.

[0034] Comparative Example 5: An oral care composition comprising, by weight percentage: 0.1% chamomile extract and 99.9% water; the chamomile extract is prepared in the same manner as in Example 1.

[0035] Comparative Example 6: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0036] The preparation method of chamomile extract is as follows: (1) After drying the chrysanthemums grown in Xinjiang, China, pass them through a 100-mesh sieve to obtain chrysanthemum powder; take 10g of chrysanthemum powder and soak it in a mixed solvent of 90mL of 1,2-propanediol and water at room temperature for 3 days with stirring; the volume ratio of 1,2-propanediol and water in the mixed solvent is 1:1. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract; the pressure during pressurization is 0.35 MPa and the pressurization time is 1 hour; the obtained chamomile extract contains 0.035 wt% total phenols and 0.000 wt% total flavonoids, of which apigenin 7-glucoside is 0.000 wt%.

[0037] Comparative Example 7: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0038] The preparation method of chamomile extract is as follows: (1) After drying the chamomile flowers grown in the Mediterranean climate region of France, pass them through a 100-mesh sieve to obtain chamomile powder; take 10g of chamomile powder and soak it in 90mL of 1,2-propanediol at room temperature for 3 days with stirring. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract; the pressure during pressurization is 0.35 MPa and the pressurization time is 1 hour; the chamomile extract obtained has a total phenol content of 0.049 wt%, a total flavonoid content of 0.038 wt%, and a flavonoid content of apigenin 7-glucoside of 0.008 wt%.

[0039] Comparative Example 8: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0040] The preparation method of chamomile extract is as follows: (1) After drying the chamomile flowers grown in the Mediterranean climate region of France, pass them through a 100-mesh sieve to obtain chamomile powder; take 10g of chamomile powder and soak it in 90mL of water at room temperature for 3 days with stirring. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract; the pressure during pressurization is 0.35 MPa and the pressurization time is 1 hour; the obtained chamomile extract contains 0.037 wt% total phenols and 0.031 wt% total flavonoids, of which the flavonoid apigenin 7-glucoside content is 0.006 wt%.

[0041] Comparative Example 9: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0042] The preparation method of chamomile extract is as follows: (1) After drying the chamomile flowers grown in the Mediterranean climate region of France, pass them through a 100-mesh sieve to obtain chamomile powder; take 10g of chamomile powder and soak it in a mixed solvent of 90mL of 1,3-propanediol and water at room temperature for 3 days with stirring; the volume ratio of 1,3-propanediol and water in the mixed solvent is 1:1. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract; the pressure during pressurization is 0.35 MPa and the pressurization time is 1 hour; the obtained chamomile extract contains 0.039 wt% total phenols and 0.032 wt% total flavonoids, of which the flavonoid apigenin 7-glucoside content is 0.006 wt%.

[0043] Comparative Example 10: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0044] The preparation method of chamomile extract is as follows: (1) After drying the whole plant of chamomile grown in the Mediterranean climate region of France, chamomile powder was obtained by passing it through a 100-mesh sieve; 10g of chamomile powder was soaked in 90mL of 1,3-propanediol at room temperature for 3 days with stirring. (2) Filtration, concentration by using vacuum rotary evaporation technology with temperature controlled at 50℃ and vacuum degree controlled at -0.08 to -0.1MPa, addition of 1.5% food-grade activated carbon and stirring at 65℃ for 45 minutes for decolorization treatment, and filtration again after decolorization to obtain the chamomile extract; the chamomile extract has a total phenol content of 0.032wt%, a total flavonoid content of 0.010wt%, and a flavonoid content of apigenin 7-glucoside of 0.002wt%.

[0045] Comparative Example 11: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0046] The preparation method of chamomile extract is as follows: (1) After drying the whole plant of chamomile grown in Xinjiang, China, pass it through a 100-mesh sieve to obtain chamomile powder; take 10g of chamomile powder and soak it in 90mL of 1,3-propanediol at room temperature for 3 days with stirring. (2) Filtration, concentration by using vacuum rotary evaporation technology to control the temperature at 50℃ and the vacuum degree at -0.08 to -0.1MPa, addition of 1.5% food-grade activated carbon and stirring at 65℃ for 45 minutes for decolorization treatment, and filtration again after decolorization to obtain the chamomile extract; the chamomile extract obtained has a total phenol content of 0.019wt%, a total flavonoid content of 0.000wt%, and a flavonoid content of apigenin 7-glucoside of 0.000wt%.

[0047] Comparative Example 12: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0048] The preparation method of chamomile extract is as follows: (1) After drying the whole plant of chamomile grown in the Mediterranean climate region of France, chamomile powder was obtained by passing it through a 100-mesh sieve; 10g of chamomile powder was soaked in 90mL of 1,2-propanediol at room temperature for 3 days with stirring. (2) Filtration, concentration by using vacuum rotary evaporation technology with temperature controlled at 50℃ and vacuum degree controlled at -0.08 to -0.1MPa, addition of 1.5% food-grade activated carbon and stirring at 65℃ for 45 minutes for decolorization treatment, and filtration again after decolorization to obtain the chamomile extract; the obtained chamomile extract has a total phenol content of 0.029wt%, a total flavonoid content of 0.008wt%, and a flavonoid content of apigenin 7-glucoside of 0.002wt%.

[0049] Comparative Example 13: An oral care composition comprising, by weight percentage: 1.5% sodium monofluorophosphate, 0.1% chamomile extract and 98.4% water.

[0050] The preparation method of chamomile extract is as follows: (1) After drying the whole plant of chamomile grown in the Mediterranean climate region of France, chamomile powder was obtained by passing it through a 100-mesh sieve. 10g of chamomile powder was soaked in a mixed solvent of 90mL of 1,2-propanediol and water at room temperature for 3 days with stirring. The volume ratio of 1,2-propanediol and water in the mixed solvent was 1:1. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract; the pressure during pressurization is 0.35 MPa and the pressurization time is 1 hour; the obtained chamomile extract contains 0.037 wt% total phenols and 0.015 wt% total flavonoids, of which the flavonoid apigenin 7-glucoside content is 0.003 wt%.

[0051] The raw material composition of the oral care compositions in the above embodiments and comparative examples is shown in Table 1.

[0052] Table 1: Composition of ingredients in oral care compositions

[0053] I. Test for inhibiting the growth of Streptococcus mutans Referring to section 7.3 of QB / T 2738-2012 "Evaluation Methods for Antibacterial and Bacteriostatic Effects of Daily Chemical Products" (Suspension Quantitative Method), the performance of the oral care compositions in the above examples and comparative examples in inhibiting the growth of Streptococcus mutans was tested. The suspension quantitative method involved mixing and culturing different antibacterial raw materials with bacterial suspensions of fixed colony counts (CFU), and calculating the antibacterial rate of the tested raw material by comparing the change in CFU.

[0054] Experimental procedure: Dilute the *Streptococcus mutans* suspension with PBS to achieve the following concentration: Add 0.1 mL of the solution to 5.0 mL of the control sample (PBS), and the recovered bacterial count should be 1 × 10⁻⁶. 4 cfu / mL ~9×10 4 CFU / mL. Dilute the test samples (Examples 1-3 and Comparative Examples 1-11) to the specified concentration with sterile standard hard water; pipette 5.0 mL of the original test sample solution into a sterile test tube and incubate at 20°C for 5 min. Pipe 0.1 mL of Streptococcus mutans bacterial solution into the test tube containing 5.0 mL of sample, mix quickly, and start timing immediately. After 2 min of incubation, take 0.5 mL of the mixture of Streptococcus mutans and the sample, add it to 4.5 mL of sterilized neutralizing agent, and mix well. After 10 min of neutralization, pipette 1 mL of the sample solution into a sterile Petri dish, inoculating two sterile Petri dishes for each sample solution or dilution. Pour 15 mL of nutrient agar medium at 40-45°C, rotate the Petri dish to ensure thorough mixing, and after the agar solidifies, invert the Petri dish. Incubate at 35±2°C for 48 hours, and then count the viable colonies. Use PBS instead of the test sample, and follow the above steps as a control sample. The experiment was repeated 3 times, and the average value was calculated. The results are shown in Table 2.

[0055] Table 2: Results of experiments on inhibiting the growth of Streptococcus mutans

[0056] in, After neutralization for 10 minutes, inoculate 1 mL of the sample solution into a sterile petri dish as an additional dilution factor in this step.

[0057] As can be seen from Table 2, the compositions prepared using the formulations of this invention in Examples 1-3 can effectively inhibit the growth of Streptococcus mutans.

[0058] Examples 1, 2, and 3 contain sodium monofluorophosphate and chamomile extract, while Comparative Example 1 does not contain sodium monofluorophosphate or chamomile extract. When comparing Examples 1, 2, 3, and Comparative Example 1, Table 2 shows that the antibacterial effect is Example 2 = Example 3 > Example 1 > Comparative Example 1, indicating that the composition containing sodium monofluorophosphate and chamomile extract can effectively inhibit the growth of Streptococcus mutans.

[0059] Comparative Examples 2 and 3 contained 0.5%-1.5% sodium monofluorophosphate but no chamomile extract. As shown in Table 2, the 0.5%-1.5% sodium monofluorophosphate content had a good antibacterial effect against Streptococcus mutans.

[0060] Comparative Examples 4 and 5 contained 0.0125%-0.1% chamomile extract but no sodium monofluorophosphate. As shown in Table 2, the antibacterial rate of Comparative Examples 4 and 5 against Streptococcus mutans was less than 50%. According to the QB / T2738-2012 standard for evaluating the antibacterial effect of antibacterial daily chemical products, the 0.0125%-0.1% chamomile extract only had a certain inhibitory effect on Streptococcus mutans but did not achieve antibacterial effect.

[0061] When comparing Examples 1-3 with Comparative Examples 2-5, Table 2 shows that Examples 1-3 had better antibacterial effects against Streptococcus mutans than the groups containing only sodium monofluorophosphate or only chamomile extract. This indicates that when the sodium monofluorophosphate content is 0.5%-1.5%, 0.0125%-0.1% chamomile extract can enhance the inhibitory effect of the composition on Streptococcus mutans, exhibiting a synergistic antibacterial effect.

[0062] The chamomile extract in Comparative Example 6 was extracted from chamomile flowers grown in Xinjiang, China. Xinjiang has a temperate continental climate with hot but unstable summers and cold, dry winters. The active ingredient content in chamomile flowers grown in Xinjiang is lower than that of chamomile flowers grown in the Mediterranean climate, which has hot, dry summers and mild, rainy winters. Table 2 shows that the antibacterial effect of the chamomile extract from the Mediterranean climate in Example 3 against *Streptococcus mutans* is better than that of the chamomile extract from Xinjiang, China in Comparative Example 6. In Comparative Examples 7-9, changing the solvent used in the chamomile extraction resulted in a decrease in the inhibitory effect of the compositions on *Streptococcus mutans* compared to Example 3. This indicates that the origin and extraction method of the chamomile significantly affect its inhibitory effect on *Streptococcus mutans*.

[0063] The chamomile extract in Comparative Example 11 was extracted from chamomile grown in Xinjiang, China. Table 2 shows that the chamomile extract in Comparative Example 10, grown in a Mediterranean climate, exhibited better inhibitory effects on *Streptococcus mutans* than the chamomile extract in Comparative Example 11, which contained chamomile grown in Xinjiang, China. This indicates that the origin of the chamomile significantly affects its inhibitory effect on *Streptococcus mutans*.

[0064] Comparative Examples 10 and 12 used chamomile extract, with only the solvent used for extraction being changed. Table 2 shows that Comparative Example 10 exhibited better inhibitory effects against *Streptococcus mutans* than Comparative Example 12. This indicates that the extraction method of chamomile significantly affects its inhibitory effect on *Streptococcus mutans*.

[0065] Comparative Example 13 used chamomile extract. As can be seen from Table 2, Example 3 has a better antibacterial effect than Comparative Example 13, indicating that the combination of chamomile extract and sodium monofluorophosphate has a better antibacterial effect than the combination of chamomile extract and sodium monofluorophosphate.

[0066] II. Test for inhibiting the growth of Porphyromonas gingivalis Referring to section 7.3 of QB / T 2738-2012 "Evaluation Methods for Antibacterial and Bacteriostatic Effects of Daily Chemical Products" (Suspension Quantitative Method), the performance of the oral care compositions in the above examples and comparative examples in inhibiting the growth of *Porphyromonas gingivalis* was tested. The suspension quantitative method involved mixing and culturing different antibacterial raw materials with bacterial suspensions containing a fixed colony count (CFU), and calculating the antibacterial rate of the tested raw material by comparing the change in CFU.

[0067] Experimental procedure: Dilute the *Porphyromonas gingivalis* suspension with PBS to achieve the following concentration: Add 0.1 mL of the solution to 5.0 mL of the control solution (PBS), and the recovered bacterial count should be 1 × 10⁻⁶. 4 cfu / mL -9×10 4 CFU / mL. Dilute the test samples (Examples 1-3 and Comparative Examples 1-11) to the specified concentration with sterile standard hard water; place 5.0 mL of the original test sample solution into a sterile test tube and incubate at 20°C for 5 min. Add 0.1 mL of *Porphyromonas gingivalis* bacterial suspension to the test tube containing 5.0 mL of sample, mix quickly, and start timing immediately. After 2 min of incubation, take 0.5 mL of the mixture of *Porphyromonas gingivalis* and the sample, add it to 4.5 mL of sterile neutralizing agent, and mix well. After 10 min of neutralization, take 1 mL of the sample solution and place it in a sterile Petri dish. Inoculate two sterile Petri dishes for each sample solution or dilution. Pour 15 mL of nutrient agar medium at 40-45°C, rotate the Petri dish to ensure thorough mixing, and invert the Petri dish after the agar solidifies. Incubate at (35±2°C) for 168 hours, and then count the viable colonies. Use PBS instead of the test sample, and follow the above steps as a control sample. The experiment was repeated 3 times, and the average value was calculated. The results are shown in Table 3.

[0068] Table 3: Results of the experiment on inhibiting the growth of Porphyromonas gingivalis

[0069] in, After neutralization for 10 minutes, inoculate 1 mL of the sample solution into a sterile petri dish as an additional dilution factor in this step.

[0070] As can be seen from Table 3, under the experimental conditions, the compositions prepared using the formulations of this invention in Examples 1-3 can effectively inhibit the growth of Porphyromonas gingivalis.

[0071] Examples 1, 2, and 3 contain sodium monofluorophosphate and chamomile extract, while Comparative Example 1 does not contain sodium monofluorophosphate or chamomile extract. When comparing Examples 1, 2, 3, and Comparative Example 1, Table 3 shows that Example 3 > Example 2 > Example 1 > Comparative Example 1, indicating that the composition containing sodium monofluorophosphate and chamomile extract can effectively inhibit the growth of Porphyromonas gingivalis.

[0072] Comparative Examples 2 and 3 contained 0.5%-1.5% sodium monofluorophosphate but no chamomile extract. As shown in Table 3, the inhibition rate of Comparative Examples 2 and 3 against Porphyromonas gingivalis was less than 50%. According to the QB / T2738-2012 standard for evaluating the antibacterial effect of antibacterial daily chemical products, the content of 0.5%-1.5% sodium monofluorophosphate has no inhibitory effect on Porphyromonas gingivalis.

[0073] Comparative Examples 4 and 5 contained 0.0125%–0.1% chamomile extract but no sodium monofluorophosphate. Table 3 shows that the 0.0125% chamomile extract only had a certain inhibitory effect on *Porphyromonas gingivalis*, but did not achieve antibacterial activity. The 0.1% chamomile extract inhibited the growth of *Porphyromonas gingivalis*. The inhibitory effect of chamomile extract on the growth of *Porphyromonas gingivalis* was dose-dependent.

[0074] When comparing Examples 1-3 with Comparative Examples 2-5, Table 3 shows that Examples 1-3 exhibited better antibacterial effects against *Porphyromonas gingivalis* than the groups containing only sodium monofluorophosphate or only matricaria champaca extract. This indicates that when the content of matricaria champaca extract is 0.0125%-0.1%, 0.5%-1.5% sodium monofluorophosphate can enhance the inhibitory effect of the composition on *Porphyromonas gingivalis*, and the two have a synergistic antibacterial effect.

[0075] The chamomile extract in Comparative Example 6 was extracted from chamomile flowers grown in Xinjiang, China. Table 3 shows that the chamomile extract from Example 3, grown in a Mediterranean climate, had a better antibacterial effect against *Porphyromonas gingivalis* than the chamomile extract from Comparative Example 6. In Comparative Examples 7-9, changing the solvent used in the chamomile extraction resulted in a decrease in the inhibitory effect of the compositions on *Porphyromonas gingivalis* compared to Example 3. This indicates that the origin and extraction method of the chamomile significantly affect its inhibitory effect on *Porphyromonas gingivalis*.

[0076] The chamomile extract in Comparative Example 11 was extracted from chamomile grown in Xinjiang, China. Table 3 shows that the chamomile extract in Comparative Example 10, grown in a Mediterranean climate, exhibited better inhibitory effects on *Porphyromonas gingivalis* than the chamomile extract in Comparative Example 11, which contained chamomile grown in Xinjiang, China. This indicates that the origin of the chamomile significantly affects its inhibitory effect on *Porphyromonas gingivalis*.

[0077] Comparative Examples 10 and 12 used chamomile extract, with only the solvent used for extraction being changed. Table 3 shows that Comparative Example 10 exhibited better inhibitory effects on *Porphyromonas gingivalis* than Comparative Example 12. This indicates that the extraction method of chamomile significantly affects its inhibitory effect on *Porphyromonas gingivalis*.

[0078] Comparative Example 13 used chamomile extract. As can be seen from Table 3, the antibacterial effect of Example 3 is better than that of Comparative Example 13, indicating that the combination of chamomile extract and sodium monofluorophosphate has a better antibacterial effect than the combination of chamomile extract and sodium monofluorophosphate.

[0079] III. Test on the effect of soothing the gums This invention utilizes the KB cell line and employs IL-1β stimulation to create an in vitro inflammation model. The soothing efficacy of the oral care compositions in the above examples and comparative examples on oral epidermal cells is verified by detecting changes in PGE2 expression levels within the system. PGE2, prostaglandin E2, is an inflammatory mediator produced from arachidonic acid under the catalysis of cyclooxygenase (COX). Acting on multiple receptors, it causes vasodilation, promotes the infiltration of neutrophils, macrophages, and mast cells from the blood to the target site, and induces symptoms such as fever, pain, and inflammation. It also acts on the nervous system to enhance pain response and is closely related to the main responses to skin irritation (redness, swelling, heat, and pain).

[0080] Experimental Methods: Negative and positive controls were set up. KB cells were seeded at an appropriate concentration in 96-well plates. After the cells reached confluence of more than 50%, the culture medium was aspirated, and the cells were washed twice with medium containing only 0.5% BSA. Then, medium containing 0.5% BSA, 10 ng / mL IL-1β, and 10% active ingredient (the concentration of the active ingredient should not be toxic to the cells) was added, and the cells were cultured for another 12 hours. The negative control was medium containing only 0.5% BSA, and the positive control was medium containing only 0.5% BSA and 10 ng / mL IL-1β, without the active ingredient. After 12 hours, 50 μL of culture medium was aspirated from each well, and the PGE2 content was measured according to the PGE2 ELISA kit instructions.

[0081] Efficacy evaluation: The results are expressed as PGE2 concentration (pg / mL). PGE2 levels were recorded in both the experimental and control groups. If the PGE2 concentration in the experimental group was significantly lower than that in the positive control, it indicated that the active ingredient had anti-inflammatory effects. The lower the PGE2 level, the stronger the anti-inflammatory effect. The results are shown in Table 4.

[0082] Table 4: Results of the Gingival Soothing Test

[0083] Wherein, #: compared with the blank control, the difference is statistically significant, p<0.05.

[0084] The difference was statistically significant compared with the control group (p<0.05).

[0085] As can be seen from Table 4, the compositions prepared using the formulations of this invention in Examples 1-3 can all significantly inhibit the expression of prostaglandin E2 (PGE2) (P<0.05).

[0086] Examples 1, 2, and 3 contain sodium monofluorophosphate and chamomile extract, while Comparative Example 1 does not contain sodium monofluorophosphate or chamomile extract. When comparing Examples 1, 2, 3, and Comparative Example 1, Table 4 shows that the PGE2 content of Example 3 < Example 2 < Example 1 < Comparative Example 1, indicating that the composition containing sodium monofluorophosphate and chamomile extract can effectively inhibit prostaglandin E2 (PGE2) expression and has a soothing effect on gingival inflammation.

[0087] Comparative Examples 2 and 3 contained 0.5%-1.5% sodium monofluorophosphate but no chamomile extract. As shown in Table 4, sodium monofluorophosphate had no effect on the expression of prostaglandin E2 (PGE2).

[0088] Comparative Examples 4 and 5 contained 0.0125%-0.1% chamomile extract but no sodium monofluorophosphate. As shown in Table 4, 0.0125%-0.1% chamomile extract effectively inhibited the expression of prostaglandin E2 (PGE2) in a dose-dependent manner.

[0089] When comparing Examples 1-3 with Comparative Examples 2-5, Table 4 shows that Examples 1-3 showed better inhibitory effects on prostaglandin E2 (PGE2) expression than the groups containing only sodium monofluorophosphate or only chamomile extract. This indicates that when the content of chamomile extract is 0.0125%-0.1%, 0.5%-1.5% sodium monofluorophosphate can enhance the inhibitory effect of the composition on prostaglandin E2 (PGE2) expression, and the two have a synergistic anti-inflammatory effect.

[0090] The chamomile extract in Comparative Example 6 was extracted from chamomile flowers grown in Xinjiang, China. Table 4 shows that the chamomile extract from Example 3, grown in a Mediterranean climate, exhibited a better inhibitory effect on prostaglandin E2 (PGE2) expression than the chamomile extract from Xinjiang, China, in Comparative Example 6. In Comparative Examples 7-9, changing the solvent used for chamomile extraction resulted in a decrease in the inhibitory effect of the compositions on prostaglandin E2 compared to Example 3. This indicates that the origin and extraction method of the chamomile significantly affect its inhibitory effect on prostaglandin E2.

[0091] The chamomile extract in Comparative Example 11 was extracted from chamomile grown in Xinjiang, China. Table 4 shows that the chamomile extract in Comparative Example 10, grown in a Mediterranean climate, had a better inhibitory effect on prostaglandin E2 (PGE2) expression than the chamomile extract in Comparative Example 11, which contained chamomile grown in Xinjiang, China. This indicates that the origin of the chamomile has a significant impact on its inhibitory effect on prostaglandin E2.

[0092] Comparative Examples 10 and 12 used chamomile extract, with only the solvent used for extraction being changed. Table 4 shows that Comparative Example 10 exhibited better inhibitory effects on prostaglandin E2 than Comparative Example 12. This indicates that the extraction method of chamomile significantly affects its inhibitory effect on prostaglandin E2.

[0093] Comparative Example 13 used chamomile extract. As can be seen from Table 4, Example 3 showed better inhibitory effect on prostaglandin E2 (PGE2) expression than Comparative Example 13, indicating that the combination of chamomile extract and sodium monofluorophosphate has a better inhibitory effect on prostaglandin E2 than the combination of chamomile extract and sodium monofluorophosphate.

Claims

1. A method for preparing a chamomile extract, characterized in that, Includes the following steps: (1) After drying the chamomile flowers grown in the Mediterranean climate, sieve them to obtain chamomile powder. Soak the chamomile powder in a mixed solvent of 1,2-propanediol and water. The volume ratio of 1,2-propanediol to water in the mixed solvent is 0.5~1.5:1, and the soaking time is 2~4 days. (2) After soaking, drain the water, then pressurize and filter to obtain the chamomile extract.

2. The method for preparing chamomile extract according to claim 1, characterized in that, In step (1), the chamomile flowers grown in the Mediterranean climate are dried and passed through an 80-100 mesh sieve.

3. The method for preparing chamomile extract according to claim 1, characterized in that, In step (1), the mass-to-volume ratio of chrysanthemum powder to mixed solvent during soaking is 1g:5~10mL.

4. The method for preparing chamomile extract according to claim 1 or 3, characterized in that, In step (1), the soaking is carried out at room temperature and stirred.

5. The method for preparing chamomile extract according to claim 1, characterized in that, In step (2), the pressure during pressurization is 0.3~0.4MPa and the pressurization time is 0.5~2h.

6. A chamomile extract, characterized in that, It is prepared using the preparation method described in any one of claims 1 to 5.

7. The chamomile extract according to claim 6, characterized in that, The total phenol content is ≥0.07% and the total flavonoid content is ≥0.06% by mass fraction.

8. The chamomile extract according to claim 6 or 7, characterized in that, The content of flavonoid apigenin-7-glucoside is ≥0.01% by mass fraction.

9. An oral care composition, characterized in that, The components include chamomile extract as described in any one of claims 4 to 6, sodium monofluorophosphate, and water.

10. The oral care composition according to claim 9, characterized in that, The components, by mass percentage, include 0.5-1.5% sodium monofluorophosphate, 0.0125-0.1% chamomile extract, and the balance being water.

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