Sugarcane polyphenol and magnolol compound raw material as well as preparation method and application thereof

Abstract

The invention discloses a sugarcane polyphenol and magnolol compound raw material as well as a preparation method and application thereof. The compound raw material is prepared from the following components in percentage by mass: 30 to 60 percent of sugarcane polyphenol extract (polyphenol is more than or equal to 20 percent), 10 to 30 percent of pure magnolol (the purity is more than or equal to 98 percent), 5 to 15 percent of cosolvent, 2 to 8 percent of stabilizer and the balance of deionized water. The preparation method comprises the following steps: extracting and purifying the sugarcane polyphenol and magnolol, dissolving the auxiliary materials, mixing, and sterilizing to obtain the finished product. The compound raw material can be used for oral care products such as toothpaste, can synergistically inhibit streptococcus mutans, destroy bacterial plaque, prevent caries and protect gingiva, is safe and stable, and is suitable for long-term use.

Description

Technical Field

[0001] This invention relates to the field of natural plant extract compounding technology, specifically to a compound raw material of sugarcane polyphenols and magnolol, as well as its preparation method and application. Background Technology

[0002] Oral diseases are among the most common human diseases. Dental caries (cavities) and gingivitis, due to their high incidence, seriously affect people's oral health and quality of life. The occurrence of dental caries is closely related to the growth, biofilm formation, and acid production of cariogenic bacteria (especially Streptococcus mutans) in the oral cavity. The acidic substances produced by Streptococcus mutans during metabolism erode tooth enamel, gradually forming cavities. Gingivitis, on the other hand, is related to the inflammatory response of the gums caused by bacterial growth in the oral cavity.

[0003] Currently, most products on the market for preventing oral diseases rely on chemically synthesized antibacterial agents (such as triclosan and chlorhexidine). While these can exert antibacterial effects in the short term, long-term use can easily lead to side effects such as oral flora imbalance, tooth staining, and altered taste, and some people may also experience allergic reactions. Therefore, developing active ingredients that are naturally derived, highly safe, have diverse mechanisms of action, and can effectively prevent oral diseases has become a research hotspot in the field of oral care.

[0004] Sugarcane polyphenols are a natural mixture of polyphenols extracted from sugarcane (Saccharum officinarum L.), containing various active ingredients such as flavonoids, phenolic acids, and anthocyanins. Existing research shows that sugarcane polyphenols have excellent antibacterial activity, especially exhibiting a significant growth-inhibiting effect on Streptococcus mutans—not only preventing the formation of Streptococcus mutans biofilm, but also specifically inhibiting its acid-producing ability, thereby reducing the erosion of teeth by acidic substances at the source and inhibiting the development of tooth decay.

[0005] Magnolia officinalis Rehd. etWils., a plant in the Magnoliaceae family, is a natural lignan compound extracted from the bark of these trees. It is the main active ingredient in magnolia officinalis. Studies have confirmed that magnolia officinalis has an inhibitory effect on various pathogenic bacteria in the oral cavity. Its anti-caries mechanism includes: inhibiting the growth and adhesion of cariogenic bacteria (such as Streptococcus mutans), disrupting the structure of existing plaque, and inhibiting the activity of Streptococcus mutans glucosyltransferase (GTF) (this enzyme is a key enzyme for bacterial synthesis of extracellular polysaccharides and biofilm formation). In addition, magnolia officinalis preparations, mouthwashes, and toothpaste products with magnolia officinalis as the active ingredient have been proven to have strong antibacterial, bactericidal, and acid-suppressing effects against various types of Streptococcus mutans, with high safety and no obvious side effects.

[0006] However, there are currently no reports on the synergistic development of active ingredients for oral care by combining sugarcane polyphenols and magnolol. When sugarcane polyphenols are used alone, their ability to destroy existing plaque is weak; when magnolol is used alone, the specificity and persistence of its inhibition of acid production by Streptococcus mutans needs improvement. Therefore, this invention combines sugarcane polyphenols and magnolol through a specific process, utilizing their complementary mechanisms of action to achieve multiple anti-caries effects of "inhibiting bacterial growth - destroying plaque - blocking acid production," while simultaneously enhancing the prevention and treatment of gingivitis, providing a new direction for the development of natural oral care ingredients. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of existing oral care raw materials, such as the large side effects of chemical antibacterial agents and the limited effects of natural single components. This invention provides a method for preparing a compound raw material of sugarcane polyphenols and magnolol. This method is simple, environmentally friendly, and the resulting compound raw material has high safety and diverse mechanisms of action, which can effectively prevent and treat oral diseases such as tooth decay and gingivitis. At the same time, this invention also discloses the application of this compound raw material in oral care products, expanding its industrialization path.

[0008] To achieve the above objectives, the present invention employs the following technical means:

[0009] A compound raw material of sugarcane polyphenols and magnolol, comprising the following components by mass percentage: 30%-60% sugarcane polyphenol extract, 10%-30% pure magnolol, 5%-15% solubilizer, 2%-8% stabilizer, and the balance being deionized water.

[0010] Preferably, the sugarcane polyphenol extract has a polyphenol content of ≥20%, the magnolol has a purity of ≥98%, the cosolvent is polyethylene glycol 400 or propylene glycol, and the stabilizer is vitamin E or tea polyphenols with a purity of ≥99%.

[0011] A method for preparing a compound raw material of sugarcane polyphenols and magnolol includes the following steps:

[0012] S1: Preparation of sugarcane polyphenol extract, the specific steps are as follows:

[0013] S11: Raw material pretreatment: Select fresh sugarcane stalks, remove leaves, roots and impurities, wash and cut into 1-2cm sections, freeze dry at -40℃~-30℃ and vacuum degree ≤10Pa for 8-12h until the moisture content is ≤5%, pulverize and pass through an 80-mesh sieve to obtain sugarcane powder;

[0014] S12: Ultrasonic-assisted extraction: Mix sugarcane powder with 70%-80% ethanol aqueous solution at a material-liquid ratio of 1:10-1:15 g / mL, and extract twice at 50℃-60℃ and ultrasonic power of 300-400W, each time for 40-60 min, and combine the extracts;

[0015] S13: Purification: After filtering the extract through a 0.45μm microporous membrane, the sample was loaded onto an AB-8 macroporous adsorption resin column. First, it was eluted with deionized water until the eluent was colorless, and then eluted with 60%-70% ethanol aqueous solution. The eluent was collected. The eluent was concentrated under reduced pressure at 50℃-60℃ and a vacuum of 0.06-0.08MPa until no alcohol odor was detected. After freeze-drying, the sugarcane polyphenol extract was obtained.

[0016] S2: Preparation or Purchase of Pure Magnoliol: If preparing, select Magnolia officinalis bark, remove impurities, and pulverize through a 60-mesh sieve to obtain Magnolia officinalis powder; mix the Magnolia officinalis powder with 95% ethanol aqueous solution at a material-to-liquid ratio of 1:8-1:12 g / mL, and reflux extract three times at 80℃-85℃, 2-3 hours each time, and combine the extracts; concentrate the extract under reduced pressure to 1 / 5 of the original volume, cool, add an equal volume of deionized water, let stand for 24 hours, filter and collect the precipitate; wash the precipitate 2-3 times with 50% ethanol aqueous solution, dry, and recrystallize twice with ethyl acetate to obtain pure magnool with a purity ≥98%; if purchasing, directly select commercially available pure magnool with a purity ≥98%;

[0017] S3: Preparation of compound raw materials:

[0018] S31: Take the amount of cosolvent in the formula, add the amount of stabilizer in the formula, and stir at 40℃-50℃ until completely dissolved to obtain a mixed excipient liquid;

[0019] S32: Add the prescribed amount of pure magnolol to the mixed excipient liquid, and stir for 30-40 minutes at 40℃-50℃ and a stirring speed of 300-500r / min until the magnolol is completely dissolved to obtain a magnolol solution.

[0020] S33: Add the formulated amount of sugarcane polyphenol extract and the remainder of deionized water to the magnolol solution, and stir for 60-80 minutes at 40℃-50℃ and a stirring speed of 500-600r / min until the system is homogeneous and transparent;

[0021] S34: The above system is sterilized by filtration through a 0.22μm microporous membrane to obtain a compound raw material of sugarcane polyphenols and magnolol.

[0022] Preferably, in step S12, the ratio of sugarcane powder to ethanol-water solution is 1:12 g / mL, the extraction temperature is 55℃, the ultrasonic power is 350W, and the extraction time is 50 min each time; in step S13, the concentration of ethanol-water solution used for elution is 65%, the vacuum concentration temperature is 55℃, and the vacuum degree is 0.07 MPa.

[0023] Preferably, in step S2, the ratio of Magnolia officinalis powder to 95% ethanol aqueous solution is 1:10 g / mL, the reflux extraction temperature is 82℃, and the extraction time is 2.5 h each time.

[0024] Preferably, in step S31, the stirring temperature is 45°C; in step S32, the stirring rate is 400 r / min and the stirring time is 35 min; in step S33, the stirring rate is 550 r / min and the stirring time is 70 min.

[0025] The application of a compound raw material of sugarcane polyphenols and magnolol in oral care products, including toothpaste, mouthwash, oral spray or tooth powder.

[0026] Preferably, the compound raw materials are used to inhibit the growth of Streptococcus mutans in the oral cavity, destroy oral plaque, inhibit the acid production of Streptococcus mutans, or to prevent and treat dental caries and gingivitis.

[0027] The present invention has the following beneficial effects:

[0028] 1. Synergistic Effect for More Significant Caries Prevention: Sugarcane polyphenols excel at inhibiting the growth of Streptococcus mutans and specifically blocking its acid production, while magnolol can destroy the structure of existing plaques and inhibit the activity of Streptococcus mutans glucosyltransferase. The combination of the two forms a triple anti-caries mechanism of "inhibiting bacterial growth - destroying plaques - blocking acid production." Experimental verification shows that the inhibitory rate of the compound raw materials against Streptococcus mutans is 20%-30% higher than that of sugarcane polyphenols alone, and the destruction rate of existing plaques is 15%-25% higher than that of magnolol alone, with a caries prevention effect far exceeding that of single ingredients.

[0029] 2. Multifunctional, combining caries prevention and gum protection: In addition to its core caries prevention function, the compound ingredients, including sugarcane polyphenols and magnolol, have anti-inflammatory activity. They can effectively inhibit the release of inflammatory factors such as IL-6 and TNF-α in the oral cavity, and relieve gingivitis symptoms such as gingival redness and bleeding. This achieves a dual oral care effect of "caries prevention + gum protection", solving the problem of the single function of traditional oral care ingredients.

[0030] 3. High safety and suitable for long-term use: All components of the compound raw materials are of natural origin or food and pharmaceutical grade excipients, and there are no side effects such as oral flora imbalance, tooth staining, and taste changes that may be caused by chemically synthesized antibacterial agents.

[0031] 4. Excellent stability and extended product shelf life: By adding vitamin E or high-purity tea polyphenols as stabilizers, the degradation of sugarcane polyphenols and magnolol due to oxidation during storage can be effectively prevented. Under accelerated storage conditions of 40℃ and 75% relative humidity, the retention rate of sugarcane polyphenols and magnolol in the compound raw materials after 6 months is ≥90%, which is much higher than that of the group without stabilizers, providing a stable guarantee for the production and storage of subsequent oral care products. Detailed Implementation

[0032] The technical solution of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] A compound raw material of sugarcane polyphenols and magnolol, comprising the following components by mass percentage: 30%-60% sugarcane polyphenol extract, 10%-30% pure magnolol, 5%-15% solubilizer, 2%-8% stabilizer, and the balance being deionized water; wherein, the polyphenol content of the sugarcane polyphenol extract is ≥20% (calculated as gallic acid), and the purity of the pure magnolol is ≥98%.

[0034] Cosolvent: Polyethylene glycol 400 (PEG-400) or propylene glycol are selected. Both are food and pharmaceutical grade excipients with high safety and can effectively improve the solubility of magnolol in water (magnolol is a fat-soluble compound and is prone to precipitation when dissolved directly in water), ensuring the uniformity and stability of the compound raw materials.

[0035] Stabilizers: Vitamin E or tea polyphenols (purity ≥99%) are selected. Both have excellent antioxidant properties, which can prevent sugarcane polyphenols and magnolol from being degraded by oxidation during storage and extend the shelf life of the compound raw materials. At the same time, vitamin E can also enhance the anti-inflammatory effect of the compound raw materials and further improve the therapeutic effect on gingivitis.

[0036] A method for preparing a compound raw material of sugarcane polyphenols and magnolol includes the following steps:

[0037] Preparation of sugarcane polyphenol extract

[0038] a. Raw material pretreatment: Select fresh sugarcane stalks, remove leaves, roots and impurities, wash and cut into 1-2cm small sections, put them into a freeze dryer, freeze dry for 8-12 hours at -40℃~-30℃ and vacuum degree ≤10Pa until the moisture content is ≤5%, then pulverize and pass through an 80-mesh sieve to obtain sugarcane dry powder.

[0039] b. Ultrasonic-assisted extraction: Mix sugarcane powder with 70%-80% ethanol aqueous solution at a material-liquid ratio of 1:10-1:15 (g / mL), place in an ultrasonic extractor, and extract twice at a temperature of 50℃-60℃ and an ultrasonic power of 300-400W, each extraction lasting 40-60 minutes. Combine the two extracts.

[0040] c. Purification: The combined extracts were filtered through a 0.45 μm microporous membrane to remove impurities. The filtrate was then loaded onto a macroporous adsorption resin (model: AB-8) column. First, deionized water was used to elute until the eluent was colorless to remove water-soluble impurities. Then, 60%-70% ethanol aqueous solution was used to elute, and the eluent was collected. The eluent was concentrated under reduced pressure at 50℃-60℃ and a vacuum of 0.06-0.08 MPa until no alcohol odor was detected, to obtain a concentrated sugarcane polyphenol solution. Finally, the concentrated solution was freeze-dried to obtain a sugarcane polyphenol extract (polyphenol content ≥20%).

[0041] Preparation of pure magnolol (optional, or commercially available pure magnolol with a purity ≥98% can be purchased directly).

[0042] a. Raw material pretreatment: Select Magnolia officinalis bark, remove impurities, and pulverize it through a 60-mesh sieve to obtain Magnolia officinalis powder.

[0043] b. Extraction: Mix Magnolia officinalis powder with 95% ethanol aqueous solution at a material-liquid ratio of 1:8-1:12 (g / mL), and reflux extract three times at 80℃-85℃, each extraction lasting 2-3 hours. Combine the extracts.

[0044] c. Purification: The extract was concentrated under reduced pressure to 1 / 5 of its original volume, cooled to room temperature, and an equal volume of deionized water was added. After stirring evenly, the mixture was allowed to stand for 24 hours to precipitate. The precipitate was collected by filtration, washed 2-3 times with a small amount of 50% ethanol aqueous solution, and dried to obtain crude magnolol. The crude product was recrystallized twice with ethyl acetate to obtain pure magnolol (purity ≥98%).

[0045] Preparation of compound raw materials

[0046] a. Dissolving the cosolvent and stabilizer: Take the amount of cosolvent (PEG-400 or propylene glycol) in the formula, add the amount of stabilizer (vitamin E or tea polyphenols) in the formula, and stir at 40℃-50℃ until completely dissolved to obtain a mixed excipient solution.

[0047] b. Dissolving magnolol: Add the prescribed amount of pure magnolol to the mixed excipient solution, and stir for 30-40 minutes at 40℃-50℃ and a stirring rate of 300-500r / min until the magnolol is completely dissolved to obtain a magnolol solution.

[0048] c. Mixing sugarcane polyphenols: Add the prescribed amount of sugarcane polyphenol extract to the magnolol solution, then add the prescribed amount of deionized water, and continue stirring for 60-80 minutes at 40℃-50℃ and a stirring rate of 500-600 r / min until the system is homogeneous and transparent with no precipitation.

[0049] d. Sterilization and packaging: The above mixture is sterilized by filtration through a 0.22μm microporous membrane to obtain a compound raw material of sugarcane polyphenols and magnolol; the compound raw material is placed into a sterile brown glass bottle, sealed and stored in a cool and dry place.

[0050] The application of a compound raw material of sugarcane polyphenols and magnolol in oral care products, including toothpaste, mouthwash, oral spray or tooth powder.

[0051] When the oral care product is toothpaste, the amount of compound ingredients added to the toothpaste is 1%-3% (by weight); the toothpaste formula also includes, by weight percentage: 35%-45% abrasive, 15%-25% humectant, 1%-3% surfactant, 1%-2% thickener, 0.05%-0.15% sweetener, 0.5%-1% fragrance, and the balance being deionized water.

[0052] The abrasive is calcium carbonate, the humectant is glycerin, the surfactant is sodium lauryl sulfate, the thickener is sodium carboxymethyl cellulose, and the sweetener is sucralose. The toothpaste formula by weight percentage is: calcium carbonate 40%, glycerin 20%, sodium lauryl sulfate 2%, sodium carboxymethyl cellulose 1.5%, sucralose 0.1%, fragrance 0.8%, compound raw materials 2%, and the balance is deionized water.

[0053] The compound ingredients are used to inhibit the growth of Streptococcus mutans in the oral cavity, destroy oral plaque, inhibit the acid production of Streptococcus mutans, or to prevent and treat dental caries and gingivitis.

[0054] Example 1

[0055] Preparation of compound raw materials of sugarcane polyphenols and magnolol

[0056] I. Preparation of Raw Materials and Auxiliary Materials

[0057] Fresh sugarcane stalks and magnolia bark;

[0058] Ethanol, polyethylene glycol 400, vitamin E;

[0059] AB-8 type macroporous adsorption resin, 0.45μm / 0.22μm microporous filter membrane.

[0060] II. Preparation Steps

[0061] Preparation of sugarcane polyphenol extract

[0062] S11: Select 5kg of fresh sugarcane stalks, remove leaves, roots and impurities, rinse with clean water 3 times and cut into 1.5cm sections, put them into a freeze dryer, freeze dry at -35℃ and vacuum degree 8Pa for 10h, and the moisture content is tested to be 4.2%; crush the dried sugarcane sections with a pulverizer and pass through an 80-mesh sieve to obtain 820g of sugarcane dry powder.

[0063] S12: Take 800g of dried sugarcane powder and mix it with 75% ethanol aqueous solution at a material-to-liquid ratio of 1:12 (g / mL). Pour the mixture into an ultrasonic extractor, set the temperature to 55℃ and the ultrasonic power to 350W, and extract twice, each time for 50 minutes. After extraction, filter the mixture with gauze to remove the residue, and combine the two extracts to obtain approximately 18000mL of extract.

[0064] S13: The combined extracts were filtered through a 0.45 μm microporous membrane to remove suspended impurities. The filtered extract was loaded onto a pre-activated AB-8 macroporous adsorption resin column at a flow rate of 2 BV / h. After loading, the column was eluted with deionized water at a flow rate of 3 BV / h, and the eluent was collected until it became colorless. Subsequently, the column was eluted with 65% ethanol aqueous solution at a flow rate of 2 BV / h, and the eluent was collected until no obvious polyphenol reaction was observed in the FeCl3 colorimetric reaction. The eluent was poured into a rotary evaporator and concentrated under reduced pressure at 55 °C and a vacuum of 0.07 MPa until no alcohol odor was detected, yielding 620 mL of sugarcane polyphenol concentrate. The concentrate was placed in a freeze dryer and freeze-dried for 12 h to obtain 128 g of sugarcane polyphenol extract. The polyphenol content was determined to be ≥20% by the Folin-Ciocalteu method.

[0065] Preparation of pure magnolol

[0066] Take 3 kg of Magnolia officinalis bark, remove impurities, cut into small pieces, pulverize using a grinder, and pass through a 60-mesh sieve to obtain 580 g of Magnolia officinalis powder. Take 500 g of Magnolia officinalis powder and mix it with 95% ethanol aqueous solution at a material-to-liquid ratio of 1:10 (g / mL). Pour the mixture into a three-necked flask and reflux extract three times under 82℃ water bath conditions, each extraction lasting 2.5 h. Combine the three extracts and concentrate them to 1000 mL using a rotary evaporator under reduced pressure at 60℃ and a vacuum of 0.08 MPa. Cool the concentrate to room temperature, add an equal volume of deionized water, stir well, and then place it on an ice bath at 4℃. After standing for 24 hours, a pale yellow precipitate was formed. The precipitate was collected by filtration using a Buchner funnel and washed three times with 50% ethanol aqueous solution. The washed precipitate was then placed in a vacuum drying oven and dried for 4 hours at 50°C and a vacuum of 0.09 MPa to obtain 42 g of crude magnolol. The crude product was added to ethyl acetate and heated to 65°C to completely dissolve it. After cooling to room temperature, the product was placed in a refrigerator at 4°C and allowed to stand for 12 hours to precipitate crystals. The crystals were collected by filtration and recrystallized once with ethyl acetate to obtain 35 g of pure magnolol, which was found to have a purity of 98.8% by high performance liquid chromatography.

[0067] Preparation of compound raw materials

[0068] S31: According to the mass percentage formula, weigh 50g of polyethylene glycol 400 and 25g of vitamin E, pour them into a beaker, place them in a 45℃ constant temperature water bath, and stir with a magnetic stirrer for 15 minutes until vitamin E is completely dissolved, to obtain 75g of mixed excipient liquid.

[0069] S32: Add 100g of pure magnolol to the mixed excipient solution, maintain the water bath temperature at 45℃, adjust the magnetic stirrer speed to 400r / min, stir for 35min, take a sample every 5min during the process, until the magnolol is completely dissolved, and obtain 175g of pale yellow transparent magnolol solution.

[0070] S33: Add 225g of sugarcane polyphenol extract to the magnolol solution, then add 100g of deionized water, maintain the temperature at 45℃, increase the stirring speed to 550r / min, and continue stirring for 70min. During this period, use a glass rod to dip a small amount of solution for observation to confirm that the system is uniform, transparent, and free of particulate precipitate.

[0071] S34: The above mixture is filtered and sterilized through a 0.22μm microporous membrane to obtain 500g of sugarcane polyphenol and magnolol compound raw material; the compound raw material is placed into a sterile brown glass bottle, sealed and labeled, and stored in a cool and dry place at 25℃.

[0072] Example 2

[0073] Application of compound ingredients in toothpaste

[0074] I. Toothpaste Formula (Total weight 1000g, percentage by weight)

[0075] Friction agent (calcium carbonate): 40% (400g);

[0076] Moisturizer (glycerin): 20% (200g);

[0077] Surfactant (sodium lauryl sulfate, SLS): 2% (20g);

[0078] Thickener (sodium carboxymethyl cellulose, CMC-Na): 1.5% (15g);

[0079] Sweetener (sucralose): 0.1% (1g);

[0080] Flavoring (mint flavoring): 0.8% (8g);

[0081] Compound raw materials (prepared in Example 1): 2% (20g);

[0082] Deionized water: Balance (336g).

[0083] II. Toothpaste preparation steps

[0084] Aqueous phase preparation: Pour 200g of glycerol and 1g of sucralose into a beaker, add 336g of deionized water, and stir with an electric stirrer (speed 300r / min) for 10min until the sucralose is completely dissolved, to obtain 537g of homogeneous aqueous solution.

[0085] Thickener dissolution: Add 15g of sodium carboxymethyl cellulose to the aqueous solution, adjust the stirring speed to 400r / min, and continue stirring for 20min. During this time, scrape off the sodium carboxymethyl cellulose residue on the beaker wall with a scraper to ensure complete dissolution. Let the solution stand for 30min to remove air bubbles, and obtain 552g of aqueous matrix.

[0086] Preparation of paste matrix: Slowly add 400g of calcium carbonate to the aqueous matrix while stirring (increase the stirring speed to 1000r / min) to prevent calcium carbonate from clumping; after the addition is complete, continue stirring for 30min to obtain 952g of white paste matrix.

[0087] Addition of compound raw materials and excipients: Reduce the stirring speed to 500 r / min, and add 20g sodium lauryl sulfate, 8g peppermint flavoring and 20g compound raw materials to the paste matrix in sequence. Stir for 5 minutes after each addition to ensure uniform mixing, and obtain 998g of toothpaste initial product (with a small loss).

[0088] Degassing and filling: Pour the initial toothpaste product into a vacuum degassing machine and degas for 15 minutes under a vacuum of 0.08 MPa. After degassing, use a toothpaste filling machine to fill the toothpaste into a tube and seal the tube opening to obtain the finished toothpaste containing compound ingredients.

[0089] Example 3:

[0090] Application of compound ingredients in mouthwash

[0091] I. Mouthwash Formula (Total weight 500g, percentage by weight)

[0092] Compound raw materials (prepared in Example 1): 1.5% (7.5g);

[0093] Glycerin (humectant, sweetener): 5% (25g);

[0094] Polysorbate 80 (surfactant, solubilizer): 0.5% (2.5g);

[0095] Menthol (flavoring): 0.1% (0.5g);

[0096] Sodium benzoate (preservative): 0.1% (0.5g);

[0097] Deionized water: Balance (463.5g).

[0098] II. Mouthwash Preparation Steps

[0099] Dissolving excipients: Pour 463.5g of deionized water into a beaker, add 25g of glycerin and 0.5g of sodium benzoate, and stir with a magnetic stirrer (speed 300r / min) for 10min until the glycerin and sodium benzoate are completely dissolved, to obtain 489g of basic solution.

[0100] Solubilization and flavoring addition: Add 0.5g of polysorbate 80 to the base solution, stir for 5 minutes, then add 0.5g of menthol and continue stirring for 15 minutes. During this time, observe the state of the solution to ensure that the menthol is completely dissolved and there is no layering. 490g of flavoring solution is obtained.

[0101] Mixing of compound raw materials: Add 7.5g of compound raw materials to the flavor solution, adjust the stirring speed to 400r / min, stir for 20min to make the compound raw materials evenly dispersed in the solution, and obtain 497.5g of initial mouthwash product.

[0102] Filtration and filling: The initial mouthwash sample is filtered through a 0.22μm microporous membrane to remove any possible tiny particles; the filtered mouthwash is then filled into sterile mouthwash bottles (250mL / bottle), sealed, and labeled to obtain the finished mouthwash product.

[0103] Specific experiments prove

[0104] Experiment 1: Antibacterial experiment of compound raw materials against oral pathogens (verifying synergistic antibacterial effect)

[0105] I. Experimental Objective

[0106] The study aimed to verify the antibacterial activity of the compound ingredients against major oral pathogens (Streptococcus mutans and Porphyromonas gingivalis), compare the antibacterial effects of sugarcane polyphenols and magnolol alone, and clarify the synergistic effect of the compound.

[0107] II. Experimental Materials

[0108] Test samples: the compound raw material prepared in Example 1 (concentration 100 μg / mL, diluted with sterile physiological saline), sugarcane polyphenol extract alone (concentration 45 μg / mL, the same content as in the compound raw material), pure magnolol alone (concentration 20 μg / mL, the same content as in the compound raw material), and sterile physiological saline (blank control).

[0109] Test strains: Streptococcus mutans (ATCC 25175, cariogenic bacterium) and Porphyromonas gingivalis (ATCC 33277, gingivitis-causing bacterium), provided by the Microbial Culture Collection Center.

[0110] Culture media: Brain Heart Infusion (BHI) medium (for culturing Streptococcus mutans) and anaerobic blood agar medium (for culturing Porphyromonas gingivalis).

[0111] Instruments: constant temperature incubator (37℃), anaerobic culture jar (including anaerobic gas generator), ELISA reader (for detecting OD600 value), sterile 96-well plate.

[0112] III. Experimental Method (Micro-broth Dilution Method)

[0113] Preparation of bacterial suspensions: *Streptococcus mutans* was inoculated onto BHI liquid medium and cultured at 37°C with 5% CO2 for 24 h; *Porphyromonas gingivalis* was inoculated onto anaerobic blood agar medium and cultured anaerobicly at 37°C for 48 h; the bacterial suspension concentrations were adjusted to 1 × 10⁻⁶ with sterile physiological saline. 6 CFU / mL.

[0114] Sample dilution: Dilute the test sample with the corresponding culture medium to 5 concentration gradients (10 μg / mL, 20 μg / mL, 40 μg / mL, 80 μg / mL, 100 μg / mL).

[0115] Antibacterial assay: In a sterile 96-well plate, add 100 μL of bacterial suspension + 100 μL of diluted sample to each well; add 100 μL of bacterial suspension + 100 μL of culture medium to each blank control well. *Streptococcus mutans* was cultured at 37℃ and 5% CO2 for 24 h, and *Porphyromonas gingivalis* was cultured anaerobically at 37℃ for 48 h. The OD600 value of each well was measured using a microplate reader (the higher the OD value, the more vigorous the bacterial growth), and the antibacterial rate was calculated.

[0116] Antibacterial rate (%) = (OD value of blank control - OD value of sample) / OD value of blank control × 100%

[0117] IV. Experimental Results

[0118] Against Streptococcus mutans, when the sample concentration was 10 μg / mL, the inhibition rate of the compound raw material was 45.2%±2.3%, the inhibition rate of the sugarcane polyphenol extract alone was 28.5%±1.8%, and the inhibition rate of pure magnolol alone was 30.1%±2.1%. When the concentration was increased to 40 μg / mL, the inhibition rate of the compound raw material reached 89.5%±2.8%, the inhibition rate of sugarcane polyphenol alone was 65.8%±3.0%, and the inhibition rate of magnolol alone was 68.2%±2.9%. When the concentration was increased to 100 μg / mL, the inhibition rate of the compound raw material was as high as 98.3%±1.5%, and the inhibition rates of sugarcane polyphenol and magnolol alone were 82.1%±2.2% and 85.7%±2.4%, respectively.

[0119] For *Porphyromonas gingivalis*, at a concentration of 10 μg / mL, the inhibitory rate of the compound raw material was 42.8% ± 2.5%, the inhibitory rate of sugarcane polyphenol alone was 25.3% ± 1.6%, and the inhibitory rate of magnolol alone was 32.6% ± 2.2%; at a concentration of 40 μg / mL, the inhibitory rate of the compound raw material was 86.7% ± 3.1%, the inhibitory rate of sugarcane polyphenol alone was 62.5% ± 2.8%, and the inhibitory rate of magnolol alone was 70.3% ± 2.6%; at a concentration of 100 μg / mL, the inhibitory rate of the compound raw material was 97.5% ± 1.8%, the inhibitory rate of sugarcane polyphenol alone was 78.9% ± 2.4%, and the inhibitory rate of magnolol alone was 88.2% ± 2.3%.

[0120] V. Results Analysis

[0121] At the same concentration, the inhibitory rates of the compound raw materials against Streptococcus mutans and Porphyromonas gingivalis were significantly higher than those of sugarcane polyphenols or magnolol alone, and the inhibitory rates showed a clear upward trend with increasing concentration. When the concentration was 100 μg / mL, the inhibitory rates of the compound raw materials against both pathogenic bacteria were close to 100%, proving that the combination of sugarcane polyphenols and magnolol produced a synergistic antibacterial effect and could effectively inhibit the growth of major oral pathogenic bacteria.

[0122] Experiment 2: Anti-caries experiment of compound raw material toothpaste (verifying clinical anti-caries effect)

[0123] I. Experimental Objective

[0124] The efficacy of toothpaste containing compound ingredients (prepared in Example 2) in preventing tooth decay was verified through human trials.

[0125] II. Experimental Materials

[0126] Experimental samples: toothpaste with compound raw materials prepared in Example 2 (experimental group) and blank toothpaste without compound raw materials (control group, other components are the same as experimental group).

[0127] Experimental population: 120 high-risk individuals with dental caries (aged 18-45 years, with 2 or more cavities in the past year, and who have not received dental treatment) were selected and randomly divided into an experimental group (60 people) and a control group (60 people). There were no significant differences in age, gender, and dental caries status between the two groups (P>0.05).

[0128] III. Experimental Methods

[0129] Instructions for use: Both groups of people used the same brushing method (Bass brushing technique), brushing their teeth twice a day for 3 minutes each time. The experimental group used compound toothpaste, while the control group used blank toothpaste. The experimental period was 12 months.

[0130] Testing indicators: Before the start of the experiment, at 6 months and 12 months of the experiment, professional dentists used the World Health Organization (WHO) dental caries diagnostic criteria to test the number of caries (DMFT index: the sum of the number of caries, missing teeth and fillings) and the degree of enamel demineralization in the two groups (detected by a laser fluorescence detector; the higher the value, the more severe the demineralization).

[0131] IV. Experimental Results

[0132] Before the experiment, the average DMFT index of the experimental group was 2.3±0.5 and the average enamel demineralization value was 15.6±2.1; the average DMFT index of the control group was 2.2±0.6 and the average enamel demineralization value was 15.8±2.3. There was no significant difference between the two groups.

[0133] Six months after the experiment, the average DMFT index of the experimental group was 2.4±0.5, only slightly increased by 0.1 compared to before the experiment; the average enamel demineralization value was 13.2±1.8, decreased by 2.4 compared to before the experiment. In contrast, the average DMFT index of the control group was 2.8±0.7, increased by 0.6 compared to before the experiment; the average enamel demineralization value was 18.5±2.5, increased by 2.7 compared to before the experiment.

[0134] After 12 months of experimentation, the average DMFT index of the experimental group was 2.5±0.6, an increase of 0.2 compared to before the experiment; the average enamel demineralization value was 12.8±1.6, a decrease of 2.8 compared to before the experiment. The average DMFT index of the control group was 3.5±0.8, an increase of 1.3 compared to before the experiment; the average enamel demineralization value was 22.3±2.8, an increase of 6.5 compared to before the experiment.

[0135] V. Results Analysis

[0136] Before the experiment, there were no significant differences in the DMFT index and enamel demineralization value between the two groups. After 6 and 12 months of the experiment, the increase in the DMFT index of the experimental group (0.2) was much lower than that of the control group (1.3), and the enamel demineralization value of the experimental group continued to decrease, while that of the control group increased significantly. This demonstrates that toothpaste containing compound ingredients can effectively inhibit the development of tooth decay, reduce enamel demineralization, and has a good clinical caries prevention effect.

[0137] Experiment 3: Gingival Protection Experiment of Compound Raw Material Mouthwash (Verifying Anti-inflammatory and Gingival Protection Effects)

[0138] I. Experimental Objective

[0139] The effect of a mouthwash containing compound ingredients (prepared in Example 3) on improving gingivitis was verified by detecting the levels of inflammatory factors and the gingival bleeding index to clarify its anti-inflammatory effect.

[0140] II. Experimental Materials

[0141] Experimental samples: the compound raw material mouthwash prepared in Example 3 (experimental group) and the blank mouthwash without compound raw material (control group, the other components are the same as the experimental group).

[0142] Experimental population: 80 patients with chronic gingivitis (aged 20-50 years, gingival bleeding index ≥2, no other oral diseases) were selected and randomly divided into an experimental group (40 people) and a control group (40 people).

[0143] Test reagents: Inflammatory factor (IL-6, TNF-α) detection kit (enzyme-linked immunosorbent assay).

[0144] III. Experimental Methods

[0145] Instructions for use: Both groups of people used the corresponding mouthwash after meals, 15mL each time, gargling for 1 minute and then spitting it out, 3 times a day, for a period of 4 weeks.

[0146] Testing indicators:

[0147] Gingival Bleeding Index (GBI): scored by a professional dentist on a scale of 0-3 (0 points: no bleeding; 3 points: severe bleeding).

[0148] Inflammatory factor levels: Gingival crevicular fluid was collected from subjects before and after the experiment, and the concentrations of IL-6 and TNF-α were detected by enzyme-linked immunosorbent assay.

[0149] IV. Experimental Results

[0150] Before the experiment, the gingival bleeding index of the experimental group was 2.4±0.3, the IL-6 concentration was 35.2±4.5 pg / mL, and the TNF-α concentration was 28.6±3.8 pg / mL; the gingival bleeding index of the control group was 2.3±0.4, the IL-6 concentration was 34.8±4.2 pg / mL, and the TNF-α concentration was 29.1±3.5 pg / mL. There was no significant difference between the two groups.

[0151] Four weeks after the experiment, the gingival bleeding index in the experimental group decreased to 0.6±0.2, a decrease of 75.0% compared to before the experiment; the IL-6 concentration decreased to 12.8±2.3 pg / mL, a decrease of 63.6% compared to before the experiment; and the TNF-α concentration decreased to 9.5±1.8 pg / mL, a decrease of 66.8% compared to before the experiment.

[0152] In the control group, the gingival bleeding index decreased to 1.8±0.3, a decrease of only 21.7% compared to before the experiment; the IL-6 concentration decreased to 28.5±3.1 pg / mL, a decrease of 18.1% compared to before the experiment; and the TNF-α concentration decreased to 23.2±2.7 pg / mL, a decrease of 20.3% compared to before the experiment.

[0153] V. Results Analysis

[0154] Four weeks into the experiment, the gingival bleeding index in the experimental group was significantly lower than that in the control group, and the decreases in IL-6 and TNF-α concentrations were far greater in the experimental group than in the control group. This indicates that the compound ingredients can effectively improve gingivitis symptoms by inhibiting the release of inflammatory factors and reducing gingival inflammation, demonstrating a clear gingival protection and anti-inflammatory effect.

[0155] The examples provided in this invention are not intended to limit the implementation. Those skilled in the art will recognize that various variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations, and any obvious variations or modifications derived therefrom are still within the scope of this invention.

Claims

1. A compound raw material of sugarcane polyphenols and magnolol, characterized in that, It is composed of the following components by mass percentage: 30%-60% sugarcane polyphenol extract, 10%-30% pure magnolol, 5%-15% solubilizer, 2%-8% stabilizer, and the balance being deionized water.

2. The method for preparing sugarcane polyphenols according to claim 1, characterized in that, The sugarcane polyphenol extract has a polyphenol content of ≥20%, the magnolol has a purity of ≥98%, the cosolvent is polyethylene glycol 400 or propylene glycol, and the stabilizer is vitamin E or tea polyphenols with a purity of ≥99%.

3. A method for preparing the compound raw material of sugarcane polyphenols and magnolol as described in claim 1 or 2, characterized in that, Includes the following steps: S1: Preparation of sugarcane polyphenol extract, the specific steps are as follows: S11: Raw material pretreatment: Select fresh sugarcane stalks, remove leaves, roots and impurities, wash and cut into 1-2cm sections, freeze dry at -40℃~-30℃ and vacuum degree ≤10Pa for 8-12h until the moisture content is ≤5%, pulverize and pass through an 80-mesh sieve to obtain sugarcane powder; S12: Ultrasonic-assisted extraction: Mix sugarcane powder with 70%-80% ethanol aqueous solution at a material-liquid ratio of 1:10-1:15 g / mL, and extract twice at 50℃-60℃ and ultrasonic power of 300-400W, each time for 40-60 min, and combine the extracts; S13： Purification: The extract was filtered through a 0.45 μm microporous membrane and loaded onto an AB-8 macroporous adsorption resin column. It was first eluted with deionized water until the eluent was colorless, and then eluted with 60%-70% ethanol aqueous solution. The eluent was collected. The eluent was concentrated under reduced pressure at 50℃-60℃ and a vacuum of 0.06-0.08 MPa until no alcohol odor was detected. After freeze-drying, the sugarcane polyphenol extract was obtained. S2: Preparation or procurement of pure magnolol: If preparing, select magnolia bark, remove impurities, and pulverize it through a 60-mesh sieve to obtain magnolia powder; mix the magnolia powder with 95% ethanol aqueous solution at a material-liquid ratio of 1:8-1:12 g / mL, and reflux extract three times at 80℃-85℃, each time for 2-3 hours, and combine the extracts; concentrate the extract under reduced pressure to 1 / 5 of the original volume, cool it, add an equal volume of deionized water, let it stand for 24 hours, filter and collect the precipitate; wash the precipitate 2-3 times with 50% ethanol aqueous solution, dry it, and recrystallize it twice with ethyl acetate to obtain pure magnolol with a purity ≥98%; If purchasing, directly select commercially available pure magnolol with a purity of ≥98%; S3: Preparation of compound raw materials: S31: Take the amount of cosolvent in the formula, add the amount of stabilizer in the formula, and stir at 40℃-50℃ until completely dissolved to obtain a mixed excipient liquid; S32: Add the prescribed amount of pure magnolol to the mixed excipient liquid, and stir for 30-40 minutes at 40℃-50℃ and a stirring rate of 300-500r / min until the magnolol is completely dissolved to obtain a magnolol solution. S33: Add the formulated amount of sugarcane polyphenol extract and the remainder of deionized water to the magnolol solution, and stir for 60-80 minutes at 40℃-50℃ and a stirring speed of 500-600r / min until the system is homogeneous and transparent; S34: The above system is sterilized by filtration through a 0.22μm microporous membrane to obtain a compound raw material of sugarcane polyphenols and magnolol.

4. The method for preparing sugarcane polyphenols according to claim 3, characterized in that, In step S12, the ratio of sugarcane powder to ethanol-water solution is 1:12 g / mL, the extraction temperature is 55℃, the ultrasonic power is 350W, and the extraction time is 50 min each time. In step S13, the concentration of ethanol-water solution used for elution is 65%, the vacuum concentration temperature is 55℃, and the vacuum degree is 0.07 MPa.

5. The method for preparing sugarcane polyphenols according to claim 3, characterized in that, In step S2, the ratio of Magnolia officinalis powder to 95% ethanol aqueous solution is 1:10 g / mL, the reflux extraction temperature is 82℃, and the extraction time is 2.5h each time.

6. The method for preparing sugarcane polyphenols according to claim 3, characterized in that, In step S31, the stirring temperature is 45℃; in step S32, the stirring rate is 400r / min and the stirring time is 35min; in step S33, the stirring rate is 550r / min and the stirring time is 70min.

7. The application of the sugarcane polyphenol and magnolol compound raw material as described in claim 1 or 2 in an oral care product, characterized in that, The oral care products include toothpaste, mouthwash, oral spray, or tooth powder.

8. The application according to claim 7, characterized in that, The compound ingredients are used to inhibit the growth of Streptococcus mutans in the oral cavity, destroy oral plaque, inhibit the acid production of Streptococcus mutans, or to prevent and treat dental caries and gingivitis.