Preparation method of composite antibacterial product

Abstract

The invention provides a preparation method of a composite antibacterial product, belongs to the technical fields of biotechnology and chemical engineering, and relates to a research, development and preparation technology of a natural antibacterial product. The compound antibacterial product disclosed by the invention adopts a unique formula of the full-spectrum sea buckthorn powder, the liquorice extract and the activated attapulgite, so that the double effects of physical adsorption and chemical bacteriostasis are realized. Wherein the whole-spectrum hippophae rhamnoides powder can inhibit the growth of bacteria, glycyrrhizic acid in the licorice extract has remarkable anti-inflammatory and antibacterial characteristics, and the activated attapulgite can effectively adsorb and remove bacteria, toxins and other harmful substances. Through the synergistic effect of the three components, the product not only has high efficiency, but also has broad-spectrum antibacterial ability.

Description

Technical Field

[0001] The present invention belongs to the technical fields of biotechnology and chemical engineering, and relates to the research and development and preparation technology of natural antibacterial products. It utilizes the natural properties of plant extracts and minerals, combines them with modern preparation technology, and develops non-traditional antibiotic antibacterial solutions for animal husbandry, and particularly relates to a preparation method of a composite antibacterial product. Background Art

[0002] In the livestock production sector, reliance on antibiotics has led to numerous problems, particularly the growth of drug resistance and the potential harm of antibiotic residues to human health and the environment. While traditional antibiotics are highly effective in controlling bacterial diseases, their long-term and excessive use has led to numerous negative impacts, including disruption of microbial balance and the development of antibiotic resistance in pathogens. This resistance is not limited to pathogens in the livestock industry but can also be transmitted to humans through the food chain, increasing the difficulty and cost of treating human diseases.

[0003] In addition, global attention to food safety and sustainable animal husbandry is growing, especially against the backdrop of the gradual restriction or ban on the use of antibiotics. For example, in July 2020, feed mills were completely banned from using antibiotics. This policy change has driven market demand for natural, non-antibiotic antibacterial solutions.

[0004] Therefore, there is an urgent need to develop a new type of natural antibacterial product, which should have high antibacterial activity and good stability, and can effectively reduce the use of traditional antibiotics without causing drug resistance. Summary of the Invention

[0005] The purpose of the present invention is to address the above-mentioned problems in the existing technology and to propose a method for preparing a composite antibacterial product.

[0006] The object of the present invention can be achieved by the following technical solutions: a method for preparing a composite antibacterial product, wherein the composite antibacterial product is formed by compounding full-spectrum seabuckthorn powder, licorice root extract powder and activated attapulgite powder; the specific steps of the preparation method of the composite antibacterial product are as follows:

[0007] S1 Preparation of full-spectrum seabuckthorn powder: S1.1 Preparation of seabuckthorn peel powder: Rinse the seabuckthorn peel with running water, dry it by microwave drying method, crush and grind the dried peel to obtain seabuckthorn peel powder; S1.2 Preparation of seabuckthorn seed powder: Use the remaining oil pressing residue of seabuckthorn seed oil prepared by cold pressing technology as raw material, dry the oil pressing residue by microwave drying method, crush and grind the dried oil pressing residue to obtain seabuckthorn seed powder; S1.3 Mixing: Evenly mix the seabuckthorn peel powder, seabuckthorn seed powder and seabuckthorn fruit powder in a certain proportion to obtain full-spectrum seabuckthorn powder; the seabuckthorn fruit powder is prepared by vacuum low-temperature belt drying process;

[0008] The reasons for choosing sea buckthorn peel powder and sea buckthorn seed powder are as follows: 1. Sea buckthorn peel: Sea buckthorn peel is rich in bioflavonoids, pectin and other ingredients. These ingredients have excellent antioxidant and anti-inflammatory effects, and bioflavonoids themselves have strong anti-inflammatory and antibacterial properties. In addition, the pectin in the peel can enhance intestinal health, help remove harmful substances and pathogens, and is suitable for improving animals' resistance to diseases, especially intestinal diseases. 2. Use the remaining oil-pressing residues of sea buckthorn seed oil as raw materials: The oil-pressing residues are sea buckthorn seed cakes. The bioactive ingredients such as flavonoids and phytosterols in sea buckthorn seed cakes have significant antioxidant and anti-inflammatory functions. These properties can help enhance the effect of composite antibacterial products, indirectly inhibiting bacterial growth by reducing inflammatory responses and free radical generation. The cellulose component of sea buckthorn seed cakes can enhance the adsorption properties of the final product, help adsorb and fix the active ingredients in licorice extract and activated attapulgite, making it easier to release when needed, thereby improving the antibacterial effect;

[0009] Antioxidant activity refers to a substance's ability to scavenge free radicals in the body or inhibit their production. Free radicals are highly reactive molecules that can cause cell damage and inflammation, potentially leading to bacterial growth and infection. Seabuckthorn peel and seed powder contain a variety of flavonoids, such as quercetin, myricetin, and rutin. These flavonoids not only have powerful antioxidant properties but are also effective free radical scavengers. In contrast, traditional supplements such as vitamins (such as C and E) and rutin, while effective antioxidants, lack the variety and quantity of the flavonoid complexes in seabuckthorn. Seabuckthorn berry powder is commercially available. Seabuckthorn berry is rich in nutrients such as vitamin C, E, and flavonoids, which are highly heat-sensitive. Traditional drying processes, while high temperatures can lead to degradation and loss of these heat-sensitive components, use vacuum low-temperature belt drying to effectively control the drying temperature, typically below 60°C, thereby maximizing the preservation of the active ingredients and nutritional value of the berry.

[0010] There are many considerations for choosing full-spectrum seabuckthorn powder over seabuckthorn berry powder alone as the raw material for a combination antibacterial product, primarily cost-effectiveness, enhanced antibacterial activity, and a comprehensive nutritional profile. Cost-effectiveness: Seabuckthorn peel and seed cake (the residual product from seabuckthorn seed oil extraction) are more cost-effective than pure seabuckthorn berry powder. Enhanced antibacterial activity: Full-spectrum seabuckthorn powder combines seabuckthorn peel and seed powder with seabuckthorn berry powder to provide a broader range of bioactive ingredients. The flavonoids and phytosterols in seabuckthorn seed cake possess significant antioxidant and anti-inflammatory properties, which contribute to the overall effectiveness of the combination antibacterial product. Flavonoids themselves possess antibacterial properties, directly inhibiting the growth of certain bacteria or indirectly by reducing inflammation and free radical production. Furthermore, components such as bioflavonoids and pectin in the seabuckthorn peel also possess anti-inflammatory and antibacterial properties, promoting intestinal health and helping to eliminate harmful substances. 3. Comprehensive Nutritional Profile: Full-spectrum seabuckthorn powder integrates the nutrients from the peel, seeds, and pulp of the seabuckthorn fruit, ensuring the product contains a variety of nutrients and bioactive ingredients from seabuckthorn. Therefore, full-spectrum seabuckthorn powder provides a cost-effective, active ingredient-rich, and nutritionally comprehensive solution for the preparation of compound antibacterial products, achieving both antibacterial and anti-inflammatory benefits while reducing costs.

[0011] The reasons for using microwave drying are as follows: The basic principle of microwave drying is to excite water molecules through electromagnetic waves (typically microwaves with a frequency of 2.45 GHz), causing them to rapidly move and generate heat. Compared to traditional heat convection or conduction methods, microwaves can act directly on the material, reducing heat loss during the drying process. This is because the water molecules are directly heated internally, eliminating the need for heat transfer to the outside through conduction. This improves efficiency and effectiveness. The risk of temperature rise is also reduced. Microwave control allows for precise temperature control, preventing the destruction of nutrients due to overheating. Experiments using four drying methods—microwave, low-temperature electric drying, natural air drying, and sun drying—resulted in the following results: A comparison of drying degree and color revealed that electric drying, even at 60°C, resulted in browning and degradation or loss of nutrients. While natural air drying and sun drying can achieve adequate drying results for sea buckthorn pomace, they are not suitable for industrial production and cannot guarantee the hygienic quality of the product. Therefore, microwave drying is preferred.

[0012] S2 Preparation of Licorice Root Extract: S2.1 First-stage Extraction: The licorice root is sequentially subjected to washing, drying, crushing, sieving, and complex enzymatic extraction to obtain a first-stage extract mixture; S2.2 Second-stage Extraction: The first-stage extract mixture is soaked in an ethanol-water solution to obtain an soaked mixture, and the soaked mixture is ultrasonically extracted using an ultrasonic device to obtain a second-stage mixture; S2.3 Concentration: The second-stage mixture is allowed to stand for a certain period of time under a constant temperature environment, and then subjected to solid-liquid separation to obtain a licorice extract; The licorice extract is concentrated using a rotary evaporator under reduced pressure to obtain a licorice concentrate; S2.4 Extract Post-processing: The licorice concentrate is sequentially subjected to vacuum low-temperature drying and grinding to obtain a licorice root extract powder;

[0013] In the composite enzyme extraction operation, the composite enzyme includes a pectinase composition and a cellulase composition, the pectinase composition includes acid endo-pectinase and acid exo-pectinase, and the cellulase composition includes acid endo-cellulase and acid exo-cellulase; disodium hydrogen phosphate-citric acid is used as a buffer; the acid endo-pectinase and acid exo-pectinase are derived from Aspergillus niger; the acid endo-cellulase and acid exo-cellulase are derived from Trichoderma reesei;

[0014] The rationale behind the selection of the combined enzyme and ultrasonic extraction methods is as follows: The pectinase and cellulase enzymes used in the combined enzyme method effectively break down the pectin and cellulose in the cell walls of licorice root roots. The properties of these enzymes allow them to selectively degrade plant cell walls, thereby releasing active ingredients within the cells (such as glycyrrhizic acid and flavonoids). The use of enzymes allows for extraction under relatively mild conditions, reducing the risk of thermal degradation of the active ingredients. These mild extraction conditions help preserve the bioactive components of licorice.

[0015] Principles of Ultrasonic Extraction: 1. Cavitation Effect: Ultrasonic extraction utilizes the cavitation effect of ultrasound to generate tiny bubbles that quickly burst. This process creates a localized high-temperature and high-pressure environment, which rapidly ruptures cell walls and further releases the active ingredients within. 2. Solubility Enhancement: Ultrasonic waves also increase the permeability of solvents, allowing them to more effectively penetrate the cell structure, enhancing the dissolution and extraction efficiency of active ingredients.

[0016] The principle behind combining complex enzyme extraction with ultrasonic extraction is as follows: After enzyme treatment, the cell walls are partially degraded, exposing the active ingredients within the cells more fully. At this point, the introduction of ultrasonic extraction can utilize the cavitation effect generated by ultrasound to further accelerate cell wall rupture and enhance the release of active ingredients. Enzyme treatment provides a good foundation for the application of ultrasound, while ultrasound can significantly enhance the extraction potential remaining after enzyme treatment.

[0017] The reasons for selecting enzyme types are as follows: Licorice root composition and characteristics: Licorice root primarily contains the following active ingredients: Glycyrrhizic acid: exhibits excellent anti-inflammatory and antibacterial properties. Flavonoids: including flavonoid glycosides and flavonoids, possess antioxidant, antibacterial, and antiviral properties. Polysaccharides and other components: exhibit immunomodulatory effects. Since most of these components are present in the cytoplasm of plant cells, their effective extraction is hampered by the cell wall. Therefore, selecting the appropriate enzyme can help effectively release these components.

[0018] Pectinase's Role: Pectin Degradation: Pectin is a key component of plant cell walls, particularly in licorice root, where it is found in the extracellular matrix. Pectinase breaks down pectin, reducing the structural strength of the cell wall and facilitating the release of cellular contents. It also promotes cell wall rupture: Pectin degradation increases cell wall permeability, making it easier to dissolve cellular contents during subsequent extraction. Reasons for choosing acid endo- and acid endo-pectinases: Acid endo- and acid endo-pectinases hydrolyze pectin molecules internally, randomly cleaving pectin chains, thereby reducing cell wall viscosity and increasing permeability. This process facilitates the extraction of active ingredients within the cell. Acid exo- and acid exo-pectinases further hydrolyze the degraded pectin chains, releasing shorter pectin chains or fragments. This further weakens the integrity of the cell wall, providing a larger surface area for subsequent extraction. Reasons for combining these two enzymes: Comprehensive extraction: The synergistic action of these two enzymes allows for more efficient release of active ingredients from licorice root, such as glycyrrhizic acid and flavonoids. Endopectinase reduces the viscosity of the cell wall by randomly cutting pectin molecules, allowing exopectinase to further degrade pectin more effectively, ultimately achieving the comprehensive extraction of more active ingredients. The structural destruction effect is obvious: endo-pectinase effectively breaks the polymer structure of pectin, greatly reducing the integrity of the cell wall; exopectinase further breaks down the already degraded pectin chains into short chains, increasing the permeability of the cell wall. This dual action allows the effective ingredients inside the cell to be dissolved and extracted more quickly. Stability of active ingredients: The enzyme activity is higher in an acidic environment, and the combination of endo- and exo-enzymes can reduce the risk of non-specific hydrolysis and the degradation of other ingredients, thereby improving the stability and purity of the active ingredients in the extract.

[0019] Regarding cellulases: Acidic endocellulases can partially hydrolyze cellulose within the cellulose chain, randomly breaking cellulose molecules to produce shorter cellulose fragments. This process significantly reduces the viscosity and stability of the cell wall, making it more susceptible to further hydrolysis and processing by subsequent exocellulases. Acidic exocellulases then gradually hydrolyze cellulose from the ends of the cellulose chain, further reducing the structural integrity of the cell wall. Exocellulases, along with endocellulases, can effectively break down already degraded cellulose fragments, releasing the embedded active ingredients of licorice. The combined use of acidic endocellulases and exocellulases can produce a synergistic effect. The endocellulases first reduce the structural integrity of the cell wall, while the exocellulases further break down the cellulose, ultimately achieving efficient extraction of the active ingredients from the licorice root. This dual action ensures the extraction of active ingredients such as flavonoids and glycyrrhizic acid in the licorice extract.

[0020] S3 Activation of Attapulgite: S3.1 Crushing, settling, filtering, drying, and preliminary grinding the attapulgite ore to obtain attapulgite particles; the drying temperature shall not exceed 120°C; S3.2 Pressurized Acid Activation: S3.2.1 First Activation Treatment: Activating the attapulgite particles with a sulfuric acid solution under pressure to obtain initial activated attapulgite; S3.2.2 Second Activation Treatment: Activating the initial activated attapulgite with a hydrochloric acid solution under pressure to obtain intermediate activated attapulgite; S3.3 Wet Grinding: After washing the intermediate activated attapulgite, the intermediate activated attapulgite is ground using a wet micro-bead mill for nanoparticle dispersion and xanthan gum as a grinding aid. After completion of the grinding operation, the activated attapulgite is washed and dried to obtain activated attapulgite powder; the drying temperature shall not exceed 120°C;

[0021] Pressure acidification offers the following advantages over conventional atmospheric pressure acidification: 1. Efficient chemical reaction kinetics: Under pressure, the intermolecular distances between the acid molecules decrease, accelerating molecular motion and increasing the frequency of intermolecular collisions. This not only accelerates the rate of the chemical reaction but also improves its completeness and efficiency. Compared to acidification under atmospheric pressure, the chemical reaction under pressure is more rapid and the conversion rate is higher, thus achieving complete activation of the attapulgite in a shorter time. 2. Improved pore structure: Under pressure, the acid molecules penetrate deeper into the tiny pores of the attapulgite. This deep penetration helps to more effectively remove impurities and existing ions, such as aluminum and iron, from the pores, thereby increasing the size and number of pores. As a result, the attapulgite treated with pressure acidification has a higher specific surface area and larger pore volume, which directly enhances its adsorption performance. 3. Reduced acid usage: Under pressure, the accelerated chemical reaction and deep acid penetration allow the required amount of acid to be significantly reduced.

[0022] By using sulfuric acid and hydrochloric acid in sequence, it is possible to effectively modify attapulgite and improve its performance. This process mainly includes several key steps such as depolymerization, cation replacement, pore dredging and surface modification. The principles are as follows: Depolymerization: Attapulgite is a layered silicate mineral with a complex layered structure. The use of sulfuric acid can cause partial depolymerization of attapulgite and destroy its interlayer structure. This depolymerization is mainly through the strong reaction of acid with metal ions such as aluminum, iron and magnesium in the mineral, resulting in partial deconstruction of the original structure. Cation replacement: Both sulfuric acid and hydrochloric acid can replace cations such as calcium and magnesium in attapulgite. During the sulfuric acid replacement process, insoluble salts such as calcium sulfate are formed. These salts remain on the surface or in the pores of the mineral, affecting its adsorption properties. The subsequent use of hydrochloric acid helps to further remove these sulfate residues left by the sulfuric acid treatment, especially calcium sulfate. Pore ​​dredging: Acid treatment, particularly hydrochloric acid treatment, not only removes insoluble sulfates from the surface and pores but also helps remove other organic and inorganic impurities. This effectively dredges the pores, increases the effective surface area of ​​the attapulgite, and improves its adsorption performance. Surface modification: Acid treatment not only removes impurities but also introduces new functional groups, such as hydroxyl groups, onto the attapulgite's surface. These new surface groups enhance its adsorption capacity for organic matter, thereby improving its overall adsorption performance.

[0023] In traditional dry grinding, the lack of a liquid medium to dissipate heat easily leads to elevated temperatures, which can cause thermal damage to the attapulgite. Dry grinding also involves strong friction and impact forces, which can damage the internal structure of the particles, such as the exfoliation of layered structures and the breakage of the crystal lattice. Dry grinding also involves relatively strong electrostatic interactions and van der Waals forces between particles, which can easily lead to particle agglomeration and affect their dispersibility. Wet grinding offers significant advantages in preparing attapulgite nanopowders, achieving finer particle sizes, better dispersibility, and more stable surface properties. The lower temperatures and friction also better protect the internal structure of the particles.

[0024] The specific steps of wet grinding are as follows: preparation stage: loading the grinding medium into the micro-bead mill, controlling the filling rate at 60%-70%, and selecting the grinding medium as cemented carbide or zirconia beads with a diameter of 0.1-0.3mm; mixing: pre-mixing the intermediate activated attapulgite with water and xanthan gum, according to the weight ratio of the intermediate activated attapulgite: water: xanthan gum is 120-130:50-60:0.25-0.35; grinding: the grinding process is divided into three stages, and a different speed is set for each stage. A lower speed (500-600rpm) is used in the initial stage to ensure that all ingredients are evenly mixed, and then gradually increased to a medium speed (such as 800-1000rpm). Finally, when approaching the desired particle size, it is increased to a high speed (1200-1500rpm) to refine the particle size.

[0025] The reasons for using xanthan gum are as follows: 1. Dispersion: Xanthan gum is a highly effective natural polymer dispersant that effectively stabilizes suspended particles in water-based media. During wet grinding, attapulgite particles are prone to agglomeration due to physical and chemical forces. Xanthan gum adsorbs onto the surface of the attapulgite particles, forming a stable colloidal protective layer that reduces direct contact and mutual attraction between particles, effectively preventing particle agglomeration and sedimentation. 2. Stability: Xanthan gum's high viscosity and colloidal properties enhance the rheological properties of the grinding slurry, providing improved suspension and flow properties. This rheological property allows for more uniform distribution of the grinding media in the mill, promoting uniform impact and abrasion between particles, thereby enhancing the grinding effect and achieving a finer particle size distribution. 3. Easy Cleaning: Xanthan gum's water solubility and biodegradability make the cleaning process simple and efficient. During the cleaning process, xanthan gum can be removed from the activated attapulgite powder by simply washing with water. Furthermore, xanthan gum's inherent environmental and non-toxic properties make post-cleaning handling and waste disposal convenient and safe.

[0026] S4 Preparation of composite slurry: S4.1 Initial mixing: Start the thermostatic container and adjust the working temperature to the preset temperature. Place the full-spectrum seabuckthorn powder and licorice root extract powder into the thermostatic container filled with purified water and stir evenly to obtain an initial mixed solution. S4.2 Intermediate mixing: Divide the activated attapulgite powder into several portions of varying weights and add the attapulgite to the initial mixed solution by a stepwise addition method to obtain an intermediate mixed solution. S4.3 Adjust the pH value: Add a pH regulating solution to the intermediate mixed solution to adjust the pH value of the mixed solution in the thermostatic container to 4-5. Continue stirring. After the stirring is completed, a composite slurry is obtained.

[0027] The pH range of 4-5 is chosen for its effectiveness: full-spectrum seabuckthorn powder is rich in organic acidic components such as polyphenols and vitamin C, which are more soluble and stable in acidic environments. Therefore, a lower pH helps release and retain the active ingredients in seabuckthorn powder. Licorice root extract contains active ingredients such as glycyrrhizic acid and flavonoids, which are more stable in acidic environments and can effectively complex with other ingredients, enhancing overall bioactivity. Activated attapulgite, as a clay mineral, alters its surface charge in acidic environments, increasing its interactions with organic molecules, such as through hydrogen bonding or electrostatic forces. To promote optimal complexation among these three materials, a slightly acidic pH range is selected. This pH range ensures the stability and solubility of the organic active ingredients in seabuckthorn powder and licorice root extract in solution. This also enhances the surface activity of the attapulgite, allowing it to more effectively bind to organic molecules, forming a stable composite structure. This adjustment significantly improves the interactions among the three components and the overall performance of the composite.

[0028] S5 slurry post-processing: the composite slurry is vacuum-dried and ground in sequence to obtain a composite antibacterial product.

[0029] Preferably, in step S1.1 and step S1.2, when drying is performed by microwave drying, the drying temperature does not exceed 60°C; in step S1.3, the full-spectrum sea buckthorn powder can pass through a 40-mesh sieve; and the weight ratio of sea buckthorn peel powder: sea buckthorn seed powder: sea buckthorn fruit powder is 8-10:5-6::5-6.

[0030] The reason for choosing a ratio of 8-10 sea buckthorn peel powder: 5-6 sea buckthorn seed powder: 5-6 sea buckthorn fruit powder is: Cost-effectiveness: Sea buckthorn peel powder and seed powder are often obtained from byproducts of sea buckthorn processing, which means using these materials can effectively reduce raw material costs. Sea buckthorn peel and seed cake (the residue after oil extraction) are relatively low-cost, while sea buckthorn fruit powder is relatively more expensive because it usually requires the entire fruit to produce. Therefore, increasing the ratio of peel powder to seed powder can reduce overall costs without sacrificing product quality. Antimicrobial and Antioxidant Activity: Sea buckthorn peel and seed powder are rich in bioactive ingredients such as flavonoids and phytosterols. These ingredients not only help inhibit bacterial growth but also have antioxidant and anti-inflammatory properties. A high ratio of these ingredients helps enhance the overall effectiveness of the antibacterial compound product. Nutritional Comprehensiveness: Full-spectrum sea buckthorn powder integrates the various nutrients of sea buckthorn, including those from the peel, seeds, and pulp. By adjusting the ratio of these three ingredients, the product not only possesses antimicrobial and anti-inflammatory properties but also provides more comprehensive nutritional support. The addition of seabuckthorn peel powder and seed powder helps enhance the product's cellulose and flavonoid content, which are particularly important for promoting intestinal health. Adsorption and ingredient release: The cellulose content of seabuckthorn seed cake enhances the product's adsorption properties, helping to adsorb and immobilize other active ingredients, such as those in licorice extract and activated attapulgite, allowing for easier release when needed. Therefore, increasing the proportion of seabuckthorn seed powder optimizes this adsorption and release mechanism. Taking all these factors into consideration, this ratio achieves the multiple goals of cost-effectiveness, enhanced antimicrobial activity, comprehensive nutritional profile, and optimized adsorption and release properties. This ratio allows each ingredient to effectively exert its role, without overdosing any one ingredient and diminishing the effectiveness of others.

[0031] Preferably, in step S2.1, the licorice root is from the taproot of licorice that is 3-4 years old; in step S2.2, the concentration of the ethanol aqueous solution is 70%; the ultrasonic equipment parameters are set as follows: the ultrasonic power is 350-400W, and the ultrasonic temperature is 40°C; in step S2.3, the rotary evaporator heating temperature is 40-60°C; in step S2.4, during the vacuum drying operation, the temperature is set to 40-60°C and the licorice root extract powder can pass through a 43-mesh sieve.

[0032] Preferably, in step S2.1, with respect to the composition of the complex enzyme, the weight ratio of the pectinase composition to the cellulase combination is 1:3; the weight ratio of acid endo-pectinase: acid exo-pectinase is 7-7.5:2.5-3; the weight ratio of acid endo-cellulase: acid exo-cellulase is 60-65:35-40; in the complex enzyme, the enzyme activity of all types is required to be ≥1×10^6U / g; the dosage of the complex enzyme is 175-177U / g·substrate; acid endo-pectinase and acid exo-pectinase are derived from Aspergillus niger; acid endo-cellulase and acid exo-cellulase are derived from Trichoderma reesei.

[0033] Acid endo-pectinase: Optimal activity temperature: 40-50°C; Optimal pH: 4.0-5.0;

[0034] Acid exopectinase: Optimal activity temperature: 45-55°C; Optimal pH: 4.5-5.5;

[0035] Acid endocellulase: Optimal activity temperature: 50-60°C; Optimal pH: 4.8-5.5;

[0036] Acid exocellulase: Optimal activity temperature: 55-65°C; Optimal pH: 5.0-6.0.

[0037] Aspergillus niger is known for its highly efficient pectinase and acidase activity, adapting to acidic conditions and crucial for extracting the pectin component of licorice. Trichoderma reesei, on the other hand, is an excellent strain for producing cellulase, effectively breaking down cellulose in plant cell walls. The enzymes produced by these two strains exhibit excellent activity and stability in acidic environments. During the licorice extraction process, the pH environment provided by the sodium hydrogen phosphate-citric acid buffer optimizes the effectiveness of these enzymes and enhances their ability to break down plant cell walls.

[0038] When selecting the combination ratio of pectinase and cellulase, there are several key factors to consider, including the cell wall composition of the target plant material, the expected extraction efficiency, and the relative activity and synergy of the various enzymes. The weight ratio of the pectinase combination to the cellulase combination is 1:3-3.5: Pectin to cellulose content ratio: There are significant differences in the pectin and cellulose content in licorice roots. Cellulose accounts for a high proportion in plant cell walls, especially in the woody part. Therefore, more cellulase is needed to effectively degrade cellulose, thereby releasing more intracellular components. Efficiency and synergy: Cellulase may take longer and have higher enzyme activity during the degradation process, so increasing its ratio can help speed up the entire extraction process. At the same time, a high ratio of cellulase can produce a better synergistic effect with pectinase, increase cell wall permeability, and thus improve overall extraction efficiency. The weight ratio of acid endo-pectinase to acid exo-pectinase is 7-7.5:2.5-3. The difference in the action of endo- and exo-pectinases: Endopectinases randomly cut within the pectin chains, significantly reducing the viscosity of the cell wall, allowing exo-pectinases to more effectively access and further degrade the pectin. A higher ratio of endo-pectinase facilitates initial cell wall degradation, providing more targets for the exo-pectinase. Extraction efficiency: A high ratio of endo-pectinase ensures extensive and uniform pectin degradation, improving the efficiency of subsequent exo-pectinase action and the release of active ingredients during extraction. The weight ratio of acid endo-cellulase to acid exo-cellulase is 60-65:35-40. Synergistic effect of cellulase: Similar to pectinases, cellulases come in both endo- and exo-acting types. Endo-cellulase randomly breaks cellulose chains, reducing overall cell wall stability and viscosity, while exo-cellulase further processes these broken cellulose fragments, promoting further cell wall decomposition. A higher ratio of endo-pectinase helps create more sites for exo-pectinase action. Cellulose stability and difficulty: Cellulose is more difficult to degrade than pectin, so a higher ratio of endonucleases is required to ensure extensive degradation of the entire cellulose, making the extraction process more efficient. In summary, the selection of a specific enzyme ratio aims to maximize extraction efficiency and fully utilize the characteristics and interactions of various enzymes to optimize the extraction of active ingredients in licorice root, such as glycyrrhizic acid and flavonoids. This ratio can effectively disrupt the cell wall structure and promote the release of more active ingredients.

[0039] The specific steps of the complex enzyme extraction operation are as follows: the licorice root and complex enzyme obtained through the screening operation are added to a constant temperature container, and then a disodium hydrogen phosphate-citric acid buffer solution with a pH value of 4.8-5.0 is added. After stirring evenly, the constant temperature container is started, the working temperature is adjusted to 50°C and the heat preservation reaction is carried out for 80-90 minutes. After the heat preservation reaction is completed, the working temperature of the constant temperature container is increased to 85-90°C, and the enzyme is inactivated for no more than 10 minutes. After the enzyme inactivation treatment is completed, the constant temperature container is closed and the first-stage extraction mixture is cooled to room temperature.

[0040] Preferably, in step S3.1, the attapulgite particles can pass through a 200-mesh sieve; in step S3.2.1, the concentration of the sulfuric acid solution is 3 mol / L; the activation treatment time is 1.5-2 hours; the pressure is 4-5 bar; the activation temperature is 70-80°C; in step S3.2.2, the concentration of the hydrochloric acid solution is 2 mol / L; the activation treatment time is 45-60 minutes; the pressure is 4-5 bar; the activation temperature is 70-80°C; in step S3.3, the particle size of the activated attapulgite powder is not higher than 30 nm.

[0041] Preferably, in step S4, the pH regulating solution is sodium hydrogen phosphate-citric acid buffer; the operating temperature of the thermostatic container is 40-45°C; and the weight ratio of full-spectrum seabuckthorn powder, licorice root extract powder, and activated attapulgite powder is 35-40:30-35:30-35. When determining the ratio, it is necessary to consider the functional properties of each ingredient and its role in the composite antibacterial product. The following is an explanation of the rationality of this configuration ratio:

[0042] The role and proportion of full-spectrum seabuckthorn powder: Functional properties: Full-spectrum seabuckthorn powder is composed of seabuckthorn peel powder, seabuckthorn seed powder, and seabuckthorn fruit powder. It is rich in vitamins C and E and various antioxidants, and has natural antibacterial and health-promoting properties. Rationale for this proportion: Full-spectrum seabuckthorn powder, comprising 35-40% of the total powder, provides sufficient active ingredients to exert its primary antibacterial and immune-enhancing functions. It is also the main component of compound antibacterial products, playing a fundamental and core role in the product's overall effectiveness.

[0043] Licorice Root Extract's Function and Ratio: Functional Properties: Licorice root extract contains flavonoids and glycyrrhizic acid, which have excellent antibacterial properties and can promote the absorption and effectiveness of other ingredients. Rationale: A 30-35% licorice root extract content ensures sufficient antibacterial potency and is sufficient to synergize and modulate the overall efficacy of the product.

[0044] Function and Ratio of Activated Attapulgite: Functional Properties: Attapulgite is a mineral with excellent adsorption properties, effectively absorbing bacteria and toxins, reducing the activity of pathogens in animals. Ratio Rationale: A 30-35% ratio ensures sufficient material to adsorb and lock in harmful substances such as bacteria and toxins. Working in conjunction with the other two ingredients, it enhances the product's overall antibacterial efficacy and safety.

[0045] Comprehensive Effects and Rational Ratio: This formulation takes into account the synergistic effects of each ingredient in antibacterial, immune-enhancing, and toxin adsorption. By balancing the amounts of each component, the efficacy, safety, and stability of the antibacterial compound are ensured. Full-spectrum seabuckthorn powder provides key nutrients and antibacterial properties, while licorice extract enhances these effects and provides additional protection, and activated attapulgite provides a physical barrier. The synergistic effect of these three ingredients in a balanced ratio optimizes the product's overall performance and meets the demand for alternatives to antibiotics in animal feed.

[0046] The pH value of disodium hydrogen phosphate-citric acid buffer can be adjusted within the range of 2.2-8.0; disodium hydrogen phosphate and citric acid can be used in the food and pharmaceutical industries and have good biocompatibility, which makes this buffer an ideal choice.

[0047] Preferably, the specific process of the intermediate mixing in step S4.2 is as follows: the activated attapulgite powder is divided into three parts of different weights, the weight of the first part accounts for 15-20% of the total weight; the weight of the second part accounts for 30-40% of the total weight, and the rest is the third part; the first part of the activated attapulgite powder is added and stirred at a speed of 100-120rpm. After the stirring is completed, ensure that the powder is fully dispersed and there is no obvious agglomeration; then add the second part of the activated attapulgite powder and stir at a speed of 150-180rpm to further promote mixing and dispersion; finally, add the third part of the activated attapulgite powder and stir at a speed of 200-250rpm until uniform.

[0048] Low-speed stirring (100-120 rpm): When adding the first portion (15-20% of the total weight), the powder is relatively small and low-speed stirring is sufficient to achieve good dispersion while preventing the powder from directly adhering to the side wall of the container due to rapid stirring.

[0049] Medium speed stirring (150-180 rpm): When adding the second portion of powder (30-40% of the total weight), increasing the stirring speed will help break up any slight agglomerates formed by the initial stirring while providing enough power to thoroughly mix the newly added powder with the already dispersed powder.

[0050] High-speed stirring (200-250 rpm): When the third portion of powder is added, the overall powder volume is already large. Using high-speed stirring can ensure uniformity throughout the mixture and avoid any local uneven powder concentration. High-speed stirring can more effectively break down the friction and adhesion between powders, promote effective mixing, and also help eliminate sedimentation caused by the weight of the materials.

[0051] Compared to existing natural antibacterial products, particularly currently used attapulgite products, the composite antibacterial product of the present invention utilizes a unique formula of full-spectrum seabuckthorn powder, licorice extract, and activated attapulgite, achieving dual benefits of physical adsorption and chemical antibacterial activity. The full-spectrum seabuckthorn powder inhibits bacterial growth, the glycyrrhizic acid in the licorice extract possesses significant anti-inflammatory and antibacterial properties, and the activated attapulgite effectively adsorbs and removes bacteria, toxins, and other harmful substances. The synergistic effect of these three ingredients ensures that this product is not only highly effective but also possesses broad-spectrum antibacterial properties. The full-spectrum seabuckthorn powder not only provides vitamins and trace elements, but also enhances its antibacterial efficacy compared to pure seabuckthorn fruit powder. This enhanced effect is primarily due to the fact that the full-spectrum seabuckthorn powder contains ingredients from seabuckthorn fruit peels and oil extraction residues, which are particularly rich in active substances such as flavonoids, alkaloids, and polyphenols. Flavonoids are powerful natural antioxidants that not only combat free radicals and reduce oxidative stress, but also possess significant antibacterial properties. Flavonoids can prevent bacterial growth and reproduction by interfering with the integrity of bacterial cell walls and cell membranes. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 This is a micrograph of the attapulgite before activation in specific example 3.

[0053] Figure 2 This is a micrograph of the activated attapulgite in specific example 3. DETAILED DESCRIPTION

[0054] The following are specific embodiments of the present invention and the accompanying drawings to further describe the technical solutions of the present invention, but the present invention is not limited to these embodiments. Specific embodiment 1:

[0056] A method for preparing a composite antibacterial product, wherein the composite antibacterial product is formed by compounding full-spectrum seabuckthorn powder, licorice root extract powder, and activated attapulgite powder; the specific steps of the composite antibacterial product preparation method are as follows:

[0057] S1 Preparation of full-spectrum seabuckthorn powder: S1.1 Preparation of seabuckthorn peel powder: Rinse the seabuckthorn peel with running water, dry it by microwave drying method, crush and grind the dried peel to obtain seabuckthorn peel powder; S1.2 Preparation of seabuckthorn seed powder: Use the remaining oil pressing residue of seabuckthorn seed oil prepared by cold pressing technology as raw material, dry the oil pressing residue by microwave drying method, crush and grind the dried oil pressing residue to obtain seabuckthorn seed powder; S1.3 Mixing: Evenly mix the seabuckthorn peel powder, seabuckthorn seed powder and seabuckthorn fruit powder in a certain proportion to obtain full-spectrum seabuckthorn powder; the seabuckthorn fruit powder is prepared by vacuum low-temperature belt drying process;

[0058] In step S1.1 and step S1.2, when drying is performed by microwave drying, the drying temperature does not exceed 60°C; in step S1.3, the full-spectrum sea buckthorn powder can pass through a 40-mesh sieve; according to the weight ratio, the sea buckthorn peel powder: sea buckthorn seed powder: sea buckthorn fruit powder is 8-10:5-6:5-6.

[0059] S2 Preparation of licorice root extract: First stage extraction: Raw material preparation: Select 3-4 year old licorice main roots. Pretreatment: Cleaning: Wash the licorice roots thoroughly. Drying: Dry the cleaned licorice roots to a humidity of no more than 5%. Crushing: Crush the dried licorice roots. Screening: Pass the crushed licorice roots through a sieve to ensure uniform particle size. Complex enzyme extraction: Prepare a complex enzyme, including a pectinase composition (acid endo-pectinase and acid exo-pectinase) and a cellulase composition (acid endo-cellulase and acid exo-cellulase), and mix them in a ratio of 1:3-3.5. The amount of synthase used is 160-170 U / g·substrate; add disodium hydrogen phosphate-citric acid buffer to adjust the pH to 4.8-5.0. Add the complex enzyme and licorice root to a constant temperature container and stir evenly. Set the temperature of the constant temperature container to 50°C and maintain the reaction for 80-90 minutes. The temperature was raised to 85-90°C for enzyme inactivation for 10 minutes. The thermostatic container was closed and cooled to room temperature to obtain the first-stage extraction mixture.

[0060] Second-Stage Extraction: Soaking Extraction: Mixture Preparation: Obtain the mixture from the first-stage extraction. Ethanol-Water Preparation: Prepare a 70% ethanol-water solution by mixing 70 liters of pure ethanol with 30 liters of deionized water. Soaking Process: Mix the first-stage extraction mixture with enough 70% ethanol-water to completely submerge the mixture. The total volume of the mixture depends on the amount of the primary extract, typically 20-22 times the weight of the extract. Soaking Time: Soak the mixture at room temperature for 30-40 minutes. Ultrasonic Extraction: Equipment Settings: Extraction is performed using an ultrasonic device with a power of 350-400 W and a frequency of 40 kHz. Ultrasonic Temperature: Maintain a temperature of 40°C during sonication to prevent loss of heat-sensitive components. Ultrasonic Time: Ultrasonic-assisted extraction is performed for 60 minutes. This duration is sufficient to enhance extraction efficiency without overheating or degrading the target compounds. Filtration and Solid-Liquid Separation: Standing: After ultrasonic extraction, the mixture is allowed to stand for 30-40 minutes to allow the solids to settle. Filtration: Use a vacuum filtration system and filter paper with appropriate pore size to remove solid residues from the soaking and extraction process.

[0061] Concentration: Rotary Evaporation: Reduced Pressure Conditions: In the rotary evaporator, set the pressure to 250 mbar. This lower pressure facilitates evaporation of the ethanol solvent at a lower temperature. Heating Temperature: Set the evaporator heating temperature to 50°C, a compromise that allows for efficient evaporation of the ethanol without excessive heating and loss of sensitive components. Concentration Time: Continue rotary evaporation until the solvent volume is reduced to 20%-30% of its original volume.

[0062] Extract post-processing: Vacuum low temperature drying: The licorice concentrate is dried under vacuum conditions at 40-60°C. Grinding: The dried material is ground to ensure that the final product can pass through a 43-mesh sieve.

[0063] S3 Activated Attapulgite: S3.1: Ore Pretreatment: Ore Crushing: Equipment: Use a jaw crusher to crush the attapulgite ore into coarse particles. Process: Ensure that all particles are crushed evenly to avoid the generation of excessively fine dust. Sedimentation and Filtration:

[0064] Equipment: Utilizes a settling tank and vacuum filtration system. Process: Crushed ore is suspended in water, allowing sedimentation to separate heavier and lighter particles. Filtration then removes fine particles and impurities. Drying and Preliminary Grinding: Drying Equipment: Hot air circulation oven.

[0065] Drying Conditions: Set the temperature no higher than 120°C to avoid structural damage. Grinding Equipment: Ball mill. Grinding Objective: Grind the dried ore to a fineness of 200 mesh, ensuring that all particles pass through a 200-mesh sieve. Step S3.2: Pressurized Acid Activation: First Activation Treatment: Activator: 3 mol / L sulfuric acid solution. Activation Equipment: Autoclave. Conditions: Temperature 70-80°C, pressure 4-5 bar, treatment time 1.5-2 hours. Procedure: In the autoclave, the attapulgite particles are mixed with sulfuric acid to ensure a thorough reaction to improve their surface properties and pore structure. Second Activation Treatment: Activator: 2 mol / L hydrochloric acid solution. Activation Equipment: Autoclave, same as above. Conditions: Temperature 70-80°C, pressure 4-5 bar, treatment time 45-60 minutes. Procedure: Hydrochloric acid is used to further improve the porosity and activity of the attapulgite and remove any sulfate residue that may have formed after the first activation.

[0066] Wet grinding: Preparation: Load the bead mill with grinding media at a fill rate of 60%-70%. Use carbide or zirconia beads with a diameter of 0.1-0.3mm. Mixing: Premix the intermediate activated attapulgite with water and xanthan gum in a weight ratio of 120-130% intermediate activated attapulgite: 50-60% water: 0.25-0.35% xanthan gum. Grinding: The grinding process is divided into three stages, with different speeds set for each stage. Initially, a lower speed (500-600 rpm) is used to ensure uniform mixing of all ingredients. The speed is then gradually increased to a medium speed (800-1000 rpm). Finally, when the desired particle size is approached, the speed is increased to a high speed (1200-1500 rpm) to refine the particle size. Refinement is performed in a wet bead mill to a particle size of no more than 30nm. Post-grinding: After grinding, the grinding liquid is filtered to remove large particles and then washed to remove any residual grinding media.

[0067] Drying: Equipment: Hot air circulation oven. Conditions: Temperature does not exceed 120℃ to ensure the stability of the particles.

[0068] S4 Preparation of Composite Slurry: Initial Mixing: Equipment Preparation: Equipment: Constant-temperature container. Temperature Setting: Adjust the operating temperature to 40-45°C to ensure effective mixing of the materials without destroying the active ingredients. Material Addition: Additives: Full-spectrum seabuckthorn powder, licorice root extract powder. Solvent: Purified water. Mixing Process: Mixing: Evenly place the above powders into a constant-temperature container containing purified water and stir evenly using a stirrer to form an initial mixture. Intermediate Mixing: Add activated attapulgite powder in steps: Quantity Division: Divide the activated attapulgite powder into three portions: 15-20% in the first portion, 30-40% in the second portion, and the remainder in the third portion. Mixing Strategy: First Portion Addition: Speed: 100-120 rpm. Objective: Ensure that the first portion of activated attapulgite powder is fully dispersed and free of noticeable agglomeration. Procedure: After adding the first portion of powder, stir to promote initial dispersion. Second Portion Addition: Speed: 150-180 rpm. Objective: Further promote mixing and dispersion. Process: After adding the second portion of powder, increase the stirring speed to improve mixing efficiency. Third addition: Speed: 200-250rpm. Goal: To achieve completely uniform mixing. According to the weight ratio, full spectrum sea buckthorn powder: licorice root extract powder: activated attapulgite powder is 35-40:30-35:30-35. pH adjustment: Add: pH adjustment liquid. Target pH range: 4-5. Process: Add an appropriate amount of pH adjustment liquid to the intermediate mixture, stir to evenly distribute and reach the target pH value. The pH adjustment liquid is disodium hydrogen phosphate-citric acid buffer; Final stirring: Stirring continues: Continue stirring until the pH value is stable and the mixture is uniform. Finished product: Obtain a composite slurry.

[0069] S5 Slurry Post-treatment: Vacuum Low-Temperature Drying Process: Equipment: Vacuum Low-Temperature Drying Equipment. Purpose: Through low-temperature drying, effectively retain the active ingredients in the composite slurry and avoid the loss of heat-sensitive components that may be caused by high temperatures. Process Parameters: Temperature Setting: The temperature is controlled at 40-45°C, ensuring that the temperature is low enough to protect sensitive ingredients. Vacuum Degree: The vacuum degree is set to 500-700 mmHg to accelerate the evaporation of moisture and reduce the potential oxidation effect of oxygen molecules in the air on the product. Drying Time: Adjusted according to the initial moisture content of the slurry. The drying time is generally expected to be 6-8 hours until the predetermined moisture content is reached, less than 5%; Operation Steps: Spread the composite slurry evenly on the drying tray, with a layer thickness not exceeding 5 mm. Place the tray in the drying equipment, start the vacuum pump, and adjust to the set temperature and vacuum degree.

[0070] Grinding: Equipment: High-efficiency micro-powder grinder. Purpose: Grind the dried material into fine powder to improve the uniformity and surface activity of the composite antibacterial product, making it easier to use and absorb. Process parameters: Grinding particle size: The target particle size is micron-level, and the particle size range is controlled within 10-20 microns. Speed: The grinding speed is set to 3000-5000rpm to ensure efficient and uniform grinding. Operation steps: Remove the dried material from the drying tray and place it into the feed hopper of the grinder. Adjust the grinder to the appropriate speed and time. The grinding time is usually 5-10 minutes. After grinding, check the uniformity of the powder through a fine sieve to ensure that no large particles remain.

[0071] Specific Example 2: The effective method for preparing liquorice root extract in step S2 was verified, and the verification process was as follows:

[0072] Materials and Preparation: Three- to four-year-old licorice root samples were collected from the same batch. Two extraction experiments were conducted: (A) using the extraction method described above; (B) using conventional hot water extraction as a control. Each experiment was repeated three times to ensure the reliability of the results. Extraction Conditions: Group A: Follow the detailed procedures in Step S2. Group B: Boil the licorice root with 10 times the volume of water for 2 hours. Active Ingredient Analysis: The content of key active ingredients, such as glycyrrhizic acid and liquiritin, was determined using high-performance liquid chromatography (HPLC). HPLC Experimental Design: Sample Preparation: Samples were prepared from the licorice root extract obtained by the combined enzymatic extraction method (Group A) and the extract obtained by conventional hot water extraction (Group B). Each sample was accurately weighed, diluted to a fixed volume with an appropriate solvent (e.g., methanol), filtered, and then used for HPLC analysis. Chromatographic Conditions: Column: C18 column (4.6 mm × 250 mm, 5 μm). Mobile phase: Water-acetonitrile gradient elution containing 0.1% formic acid. Flow rate: 1.0 mL / min. Column temperature: Room temperature. Detector: UV detector, detection wavelength set at 254 nm. Standard curve: Prepare a standard curve using glycyrrhizic acid and liquiritin standards of known concentrations to ensure the accuracy of quantitative analysis. Sample injection volume: Inject 20 μL of sample into the HPLC system at a time.

[0073] Data collection and display:

[0074] Sample number Extraction Group Glycyrrhizic acid content (mg / g) Liquiritin content (mg / g) 1 Group A 10.2 15.5 2 Group A 10.5 15.8 3 Group A 10.3 15.3 1 Group B 5.8 8.0 2 Group B 5.9 8.2 3 Group B 5.7 7.9

[0075] Taking glycyrrhizic acid as an example, the gradient elution process of a sample with a glycyrrhizic acid content of 10.5 mg / g in high performance liquid chromatography (HPLC) is as follows: HPLC gradient elution data table

[0076]

[0077] Data analysis: Glycyrrhizic acid content: Group A: The average content was 10.33 mg / g, and the standard deviation was 0.15 mg / g.

[0078] Group B: Average content was 5.80 mg / g, with a standard deviation of 0.10 mg / g. Liquiritin content: Group A: Average content was 15.53 mg / g, with a standard deviation of 0.25 mg / g. Group B: Average content was 8.03 mg / g, with a standard deviation of 0.15 mg / g. These data indicate that the extraction effect of Group A was significantly better than that of Group B, with both glycyrrhizic acid and glycyrrhizin contents significantly higher than those of Group B. The relatively low standard deviation of Group A indicates good consistency and repeatability in its extraction process.

[0079] Specific embodiment 3: Figure 1 and Figure 2 As shown, the attapulgite is activated by step S3 of specific embodiment 1, and the specific process is as follows:

[0080] S3.1: Ore Pretreatment: Ore Crushing: Equipment: Use a jaw crusher to crush the attapulgite ore into coarse particles. Process: Ensure uniform crushing of all particles to avoid the generation of excessively fine dust. Sedimentation and Filtration: Equipment: Utilize a settling tank and vacuum filtration system. Process: The crushed ore is suspended in water to separate the heavier and lighter particles by sedimentation, and then filtered to remove fine particles and impurities. Drying and Preliminary Grinding: Drying Equipment: Hot air circulation oven.

[0081] Drying Conditions: Set the temperature to 106°C to avoid structural damage. Grinding Equipment: Ball mill. Grinding Objective: Grind the dried ore to a fineness of 200 mesh, ensuring that all particles pass through a 200-mesh screen. Step S3.2: Pressurized Acid Activation: First Activation Treatment: Activator: 3 mol / L sulfuric acid solution. Activation Equipment: Autoclave. Conditions: Temperature 80°C, Pressure 5 bar, Treatment Time 2 hours. Procedure: In the autoclave, the attapulgite particles are mixed with sulfuric acid to ensure a sufficient reaction to improve their surface properties and pore structure. Second Activation Treatment: Activator: 2 mol / L hydrochloric acid solution. Activation Equipment: Autoclave, same as above. Conditions: Temperature 80°C, Pressure 5 bar, Treatment Time 60 minutes. Procedure: Hydrochloric acid is used to further improve the porosity and activity of the attapulgite and remove any sulfate residue that may have formed after the first activation.

[0082] Wet Grinding: Preparation: Load the bead mill with grinding media at a 67% fill rate, using zirconia beads with a diameter of 0.1 mm. Mixing: Premix the intermediate activated attapulgite with water and xanthan gum in a weight ratio of 125:60:0.3. Grinding: The grinding process is divided into three stages, with different speeds set for each stage. Initially, a low speed of 600 rpm is used to ensure uniform mixing of all ingredients. This is then gradually increased to a medium speed of 900 rpm. Finally, when the desired particle size is approached, a high speed of 1200 rpm is used to refine the particle size. Refine the particles in a wet bead mill to a particle size of no more than 30 nm. Post-Grinding: After grinding, the grinding liquid is filtered to remove large particles and then washed to remove any residual grinding media.

[0083] Drying: Equipment: Hot air circulation oven. Conditions: Temperature is 106°C to ensure the stability of the particles.

[0084] After comparison Figure 1 and Figure 2 The original material exhibited a dense surface structure, with its crystals clustered into bundles in the form of rods or fibers, then randomly stacked into plate-like or agglomerated structures. This dense and orderly aggregation resulted in poor dispersibility in applications, as the tightly packed crystals reduced the surface area and accessible active sites. The density of this structure resulted in a smaller surface area available for adsorption. The limited inter-crystal spacing and low porosity made it difficult for adsorbed molecules to penetrate deeply into the material, limiting its adsorption capacity and rate.

[0085] The surface morphology of the activated material undergoes significant changes. The crystal bundles become more elongated and loose. This structural shift leads to the appearance of more spaces between the crystals, forming open pores. These pores not only increase the material's porosity but also add to the surface irregularity and complexity. These pores provide more surface area and higher pore connectivity. Furthermore, the presence of pores facilitates molecular diffusion and penetration, increasing adsorption rate and capacity.

[0086] Third-party testing: The composite antibacterial product prepared in Example 1 was sent to a third-party testing agency for testing. Referring to Part II of the Technical Specifications for Disinfection (Ministry of Health, 2002 edition), the test results are as follows:

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095] The specific embodiments described herein are merely illustrative of the present invention. Persons skilled in the art may make various modifications, additions, or substitutions to the described embodiments without departing from the spirit of the present invention or exceeding the scope thereof. While the present invention has been described and illustrated in detail in the accompanying drawings and the foregoing description, such illustration and description are to be considered illustrative or exemplary rather than restrictive.

Claims

1. A method for preparing a composite antibacterial product, characterized in that: The composite antibacterial product is formed by compounding full-spectrum seabuckthorn powder, licorice root extract powder and activated attapulgite powder; the specific steps of the preparation method of the composite antibacterial product are as follows: S1 Preparation of full-spectrum seabuckthorn powder: S1.1 Preparation of seabuckthorn peel powder: Rinse the seabuckthorn peel with running water, dry it by microwave drying method, crush and grind the dried peel to obtain seabuckthorn peel powder; S1.2 Preparation of seabuckthorn seed powder: Use cold pressing technology to prepare seabuckthorn seed oil. The remaining oil pressing residue is used as raw material, and the oil pressing residue is dried by microwave drying method. The dried oil pressing residue is crushed and grinded to obtain seabuckthorn seed powder; S1.3 Mixing: Evenly mix seabuckthorn peel powder, seabuckthorn seed powder and seabuckthorn fruit powder in a certain proportion to obtain full-spectrum seabuckthorn powder; the seabuckthorn fruit powder is prepared by vacuum low-temperature belt drying process; S2 Preparation of licorice root extract: S2.1 First stage extraction: The licorice root is sequentially subjected to washing, drying, crushing, screening and composite enzyme extraction operations to obtain a first stage extraction mixture; S2.2 Second stage extraction: The first stage extraction mixture is placed in an ethanol aqueous solution for soaking to obtain an soaking mixture, and the soaking mixture is subjected to ultrasonic extraction by ultrasonic equipment to obtain a second stage mixture; S2.3 Concentration: The second stage mixture is placed under a constant temperature environment, and then subjected to solid-liquid separation to obtain a licorice extract; The licorice extract is concentrated using a rotary evaporator under reduced pressure to obtain a licorice concentrate; S2.4 Extract post-processing: The licorice concentrate is sequentially subjected to vacuum low-temperature drying and grinding to obtain a licorice root extract powder; In the composite enzyme extraction operation, the composite enzyme includes a pectinase composition and a cellulase composition, the pectinase composition includes acid endo-pectinase and acid exo-pectinase, and the cellulase composition includes acid endo-cellulase and acid exo-cellulase; Disodium hydrogen phosphate-citric acid was used as the buffer; Acid endo-pectinase and acid exo-pectinase are derived from Aspergillus niger; Acid endocellulase and acid exocellulase were derived from Trichoderma reesei; S3 Activated attapulgite: S3.1 Pre-treat the attapulgite ore to obtain attapulgite particles; S3.2 Pressurized acid activation: S3.2.1 First activation treatment: Use sulfuric acid solution under pressure to activate the attapulgite particles to obtain initial activated attapulgite; S3.2.2 Second activation treatment: Use hydrochloric acid solution under pressure to activate the initial activated attapulgite to obtain intermediate activated attapulgite; S3.3 Wet grinding: Use a wet micro-bead mill for nanoparticle dispersion as a grinding equipment and select xanthan gum as a grinding aid to grind the intermediate activated attapulgite. After the grinding operation is completed, wash and dry it to obtain activated attapulgite powder; S4 Preparation of composite slurry: S4.1 Initial mixing: Start the thermostatic container, adjust the working temperature to the preset temperature, put the full spectrum seabuckthorn powder and licorice root extract powder into the thermostatic container filled with purified water, and stir evenly to obtain an initial mixed solution; S4.2 Intermediate mixing: Divide the activated attapulgite powder into several portions of different weights, and add the attapulgite into the initial mixed solution by a step-by-step addition method; obtain an intermediate mixed solution; S4.3 Adjust the pH value: Add a pH value adjusting liquid to the intermediate mixed solution, and adjust the pH value of the mixed solution in the constant temperature container to 4-5; then continue stirring, and after the stirring is completed, obtain a composite slurry; S5 slurry post-treatment: The composite slurry is vacuum-dried and ground in turn to obtain a composite antibacterial product.

2. The method for preparing the composite antibacterial product according to claim 1, characterized in that: In step S1.1 and step S1.2, when drying is performed by microwave drying, the drying temperature does not exceed 60°C; in step S1.3, the full-spectrum seabuckthorn powder can pass through a 40-mesh sieve; in terms of weight ratio, the seabuckthorn peel powder: seabuckthorn seed powder: seabuckthorn fruit powder is 8-10:5-6:5-6.

3. The method for preparing the composite antibacterial product according to claim 1, characterized in that: In step S2.1, the licorice root comes from the taproot of licorice with a year of 3-4 years; in step S2.2, the concentration of the ethanol aqueous solution is 70%; the ultrasonic equipment parameters are set as follows: the ultrasonic power is 350-400W, and the ultrasonic temperature is 40°C; in step S2.3, the rotary evaporator heating temperature is 40-60°C; in step S2.4, in the vacuum drying operation, the temperature is set to 40-60°C, and the licorice root extract powder can pass through a 43-mesh sieve.

4. The method for preparing the composite antibacterial product according to claim 1, characterized in that: In step S2.1, with respect to the composition of the composite enzyme, the weight ratio of the pectinase composition to the cellulase composition is 1:3-3.5; the weight ratio of acid endo-pectinase: acid exo-pectinase is 7-7.5:2.5-3; the weight ratio of acid endo-cellulase: acid exo-cellulase is 60-65:35-40; in the composite enzyme, the enzyme activity of all types is required to be ≥1×10^6U / g; the dosage of the composite enzyme is 160-170U / g·substrate.

5. The method for preparing the composite antibacterial product according to claim 1, characterized in that: In step S2.1, the specific steps of the complex enzyme extraction operation are as follows: add the licorice root and the complex enzyme obtained by the screening operation into a constant temperature container, then add disodium hydrogen phosphate-citric acid buffer with a pH value of 4.8-5.0, stir evenly, start the constant temperature container, adjust the working temperature to 50°C for insulation reaction for 80-90 minutes, after the insulation reaction is completed, increase the working temperature of the constant temperature container to 85-90°C, and perform enzyme inactivation treatment, the enzyme inactivation treatment time does not exceed 10 minutes, after the enzyme inactivation treatment is completed, close the constant temperature container, and cool the obtained first stage extraction mixture to room temperature.

6. The method for preparing the composite antibacterial product according to claim 1, characterized in that: In step S3.1, the process of pre-treating the attapulgite ore includes crushing, sedimentation, filtration, drying and preliminary grinding; the attapulgite particles can pass through a 200-mesh sieve; in step S3.2.1, the concentration of the sulfuric acid solution is 3 mol / L; the activation treatment time is 1.5-2 hours; the pressure is 4-5 bar; the activation temperature is 70-80°C; in step S3.2.2, the concentration of the hydrochloric acid solution is 2 mol / L; the activation treatment time is 45-60 minutes; the pressure is 4-5 bar; the activation temperature is 70-80°C; in step S3.3, the particle size of the activated attapulgite powder is not higher than 30 nm, and in steps S3.1 and S3.3, the drying temperature does not exceed 120°C.

7. The method for preparing the composite antibacterial product according to claim 1, characterized in that: In step S4, the working temperature of the thermostatic container is 40-45°C; the weight ratio of full-spectrum seabuckthorn powder: licorice root extract powder: activated attapulgite powder is 35-40:30-35:30-35, and the pH regulating solution is disodium hydrogen phosphate-citric acid buffer.

8. The method for preparing the composite antibacterial product according to claim 1, characterized in that: The specific process of intermediate mixing in step S4.2 is as follows: the activated attapulgite powder is divided into three portions of different weights, the weight of the first portion accounts for 15-20% of the total weight; the weight of the second portion accounts for 30-40% of the total weight, and the rest is the third portion; the first portion of activated attapulgite powder is added, and stirring is performed at a speed of 100-120rpm. After stirring, ensure that the powder is fully dispersed and there is no obvious agglomeration; then add the second portion of activated attapulgite powder, and stir at a speed of 150-180rpm to further promote mixing and dispersion; finally, add the third portion of activated attapulgite powder, and stir at a speed of 200-250rpm until uniform.

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