Herbal composition for nasal cavity nursing and preparation method thereof

Abstract

The invention provides a herbal composition for nasal cavity nursing and a preparation method thereof, and belongs to the technical field of health care products. The composition comprises sweet potato vinegar primary pulp, extracts of various herbaceous plants such as fructus xanthii, flos magnoliae liliflorae and radix angelicae, and auxiliary materials such as garlic and honey. The preparation method comprises the steps of raw material pretreatment, step-by-step dynamic countercurrent extraction, flavor and thermosensitive component integrated extraction, precise separation and refining, controllable homogenization and stabilization, sterile filling and the like. Through cooperation of multiple components, uncomfortable symptoms such as nasal cavity dryness, tightness and unsmooth ventilation can be comprehensively relieved; according to the process, the extraction efficiency of active ingredients and the product stability are improved; the composition disclosed by the invention is mild, non-irritant to nasal mucosa and high in safety.

Description

Technical Field

[0001] This invention relates to the field of health product technology, and in particular to an herbal composition for nasal care and its preparation method. Background Technology

[0002] As the primary gateway to the respiratory system, the health of the nasal cavity directly affects breathing comfort and local immune defense. Nasal dryness, tightness, and abnormal secretions are common discomforts affecting the quality of life for a wide range of people. Currently, there are various products and technological solutions for nasal care, but existing technologies still have many limitations in terms of formulation and implementation.

[0003] In existing technologies, nasal care products mainly fall into the following categories: First, nasal drops or sprays primarily composed of chemically synthesized ingredients, such as saline solution, seawater spray, or preparations containing decongestants or antihistamines. While these products may have a relatively quick onset of action, long-term use of chemical drugs may lead to risks such as mucosal dryness, drug-induced rhinitis, or drug resistance. Furthermore, their effects are often singular, lacking a comprehensive conditioning effect on the nasal environment. Second, traditional preparations based on single or a few Chinese medicinal herbs, such as some Chinese herbal nasal drops or medicated oils. Although these products utilize certain characteristics of natural herbs, their simple formulations and relatively isolated functions of each component make it difficult to achieve a synergistic and complex care effect. They also have limited effectiveness in improving the user experience and alleviating various complex discomforts.

[0004] Invention Patent Content In view of this, the present invention aims to provide an herbal composition for nasal care and a method for preparing the same, in order to solve or alleviate the technical problems existing in the prior art.

[0005] The technical solution of this invention patent embodiment is implemented as follows: A herbal composition for nasal care, by weight fraction, includes the following ingredients: 800-1200 parts sweet potato vinegar paste, 12-18 parts cocklebur, 6-10 parts magnolia flower, 4-8 parts angelica, 3-5 parts perilla leaf, 25-35 parts garlic, 4-6 parts Sichuan pepper, 4-6 parts sesame oil, 5-7 parts honey, 2-4 parts ginger, 2-4 parts scallion, 4-6 parts sophora flavescens, and 0.8-1.2 parts charcoal.

[0006] As an improvement, the following ingredients are included by weight fraction: 1000 parts sweet potato vinegar base, 15 parts cocklebur, 8 parts magnolia flower, 6 parts angelica, 4 parts perilla leaf, 30 parts garlic, 5 parts Sichuan pepper, 5 parts sesame oil, 6 parts honey, 3 parts ginger, 3 parts scallion, 5 parts sophora flavescens, and 1 part charcoal.

[0007] A method for preparing an herbal composition for nasal care includes the following steps: S1. Raw material pretreatment: Xanthium sibiricum, Magnolia biondii, Angelica dahurica, and Sophora flavescens are cleaned, microwave-dried to a moisture content of ≤8%, then pulverized using a universal pulverizer and passed through an 80-mesh sieve to obtain herbal powder; Perilla frutescens leaves, ginger, and scallion whites are washed, sterilized with ozone water, and then cut into 5±1mm segments to obtain fresh materials for use; Garlic is peeled and washed, and then pulverized with Sichuan peppercorns using a low-temperature airflow pulverizer to a particle size D90≤150μm to obtain garlic micro powder and Sichuan peppercorn micro powder; Charcoal is ultra-finely pulverized under inert gas protection and passed through an 800-mesh sieve to obtain ultra-fine charcoal powder, which is then placed in a moisture-proof container for later use; S2. Stepwise dynamic countercurrent extraction: The herbal powder obtained in step S1 and all the sweet potato vinegar pulp were put into a multi-functional hot reflux extraction tank. Stirring was turned on, and the temperature was raised to 50±2℃ at a rate of 1.5℃ / min. The temperature was maintained and ultrasonically assisted extraction was performed at a power density of 50W / L for 45min. The temperature was then raised to 68±2℃ at a rate of 1℃ / min. Dynamic countercurrent circulation extraction was performed at this temperature for 4 hours. During the extraction process, a slight negative pressure of -0.02 to -0.03MPa was maintained in the tank. After the extraction was completed, the liquid was cooled to below 40℃ through the built-in cooling coil to obtain the primary alcohol extract. S3. Integrated extraction of flavor and heat-sensitive components: The fresh material to be used, garlic powder and Sichuan pepper powder obtained in step S1 are added together to the primary alcohol extract and warm-soaked for 90 min at 60±2℃ and under sealed conditions with a stirring speed of 120 rpm. S4. Precision separation and purification: The mixture after step S3 is fed into a tubular centrifuge and solid-liquid separation is performed under a centrifugal force of 10000G. The supernatant is collected. The supernatant is then filtered sequentially through a 50μm bag filter and a 5μm precision filter to obtain a clear extract. S5. Controllable homogenization and stabilization: The clarified extract is introduced into a jacketed cooling mixing tank, cooled and maintained at 25±3℃; under continuous stirring at 80rpm / min, sesame oil and honey are slowly and evenly added to the tank, and stirred for 20min until completely emulsified and dispersed; with the high-speed shear emulsifier running at 10000rpm / min, the ultrafine charcoal powder obtained in step S1 is evenly fed into the emulsifier head through a vacuum feeding device, and homogenized continuously for 15min to form a stable suspension system; S6. Aseptic filling: The homogenized semi-finished product is filtered through a 0.45μm terminal sterilization filter and then transferred to an aseptic temporary storage tank; using a light-proof and well-sealed pharmaceutical nasal drop bottle or spray bottle, the product is filled and capped in a Class B clean environment to obtain the finished herbal composition for nasal care.

[0008] As an improvement, the dynamic countercurrent circulation extraction in step S2 is achieved by pumping the liquid from the bottom of the extraction tank to the spray device on the top of the tank to form a continuous internal circulation, with a circulation flow rate of 2-3 times the tank volume per hour.

[0009] As an improvement, in step S4, after the tubular centrifugation, the obtained solid residue is rinsed with 30% of its mass of pure water. The rinsing liquid is collected separately, concentrated at low temperature, and then added back to the supernatant to improve the transfer rate of the target component.

[0010] As an improvement, in step S5, the sesame oil is subjected to nitrogen bubbling deoxygenation treatment for 10 minutes before use, and the honey is filtered through a 40-mesh sieve to remove crystallized particles and impurities.

[0011] As an improvement, in step S5, the ultrafine charcoal powder is premixed with an equal weight of clarified extract to form a slurry before being inhaled, and then quantitatively delivered to the emulsifying head by a peristaltic pump.

[0012] As an improvement, in step S6, before filling, high-purity nitrogen is introduced into the sterile temporary storage tank to replace the air at the top of the tank for 10 minutes, and the tank is kept at a slightly positive nitrogen pressure of 0.03-0.05 MPa until filling is completed.

[0013] The embodiments of this invention, employing the above technical solutions, possess the following advantages: Through the synergistic design of a multi-component herbal formula, using sweet potato vinegar as a matrix, it integrates herbal ingredients such as Xanthium sibiricum, Magnolia biondii, and Angelica dahurica, as well as natural excipients such as garlic and Sichuan pepper, forming a comprehensive conditioning effect on multiple discomforts such as nasal dryness, tightness, and poor ventilation. This overcomes the shortcomings of existing chemical preparations, which have a single effect and are prone to causing mucosal dependence or damage, as well as the limitations of traditional Chinese medicine formulas, which are simple and have isolated functions of each component, making synergistic effects difficult. The advanced preparation process, employing stepwise dynamic countercurrent extraction, flavor integration extraction, precise separation, and controllable homogenization, significantly improves the extraction efficiency and transfer rate of active ingredients, ensuring product uniformity, stability, and shelf-life quality, and avoiding component loss, easy stratification, and oxidative deterioration problems caused by traditional static soaking processes. The composition of this invention has excellent local tolerance and mildness, is non-toxic to nasal mucosal cells, and can regulate microbial growth and biofilm formation, improving user comfort while ensuring safety, thus achieving a balance between efficacy, safety, and stability in nasal care products.

[0014] The above overview is for illustrative purposes only and is not intended to be limiting in any way. Further aspects, embodiments, and features of this invention will become readily apparent from the following detailed description, in addition to the illustrative aspects, embodiments, and features described above. Detailed Implementation

[0015] The invention will be more readily understood by referring to the following detailed description of preferred embodiments and included examples. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of conflict, the definitions in this specification shall prevail.

[0016] As used herein, the terms “prepared from” and “comprising” are synonymous. The terms “comprising,” “including,” “having,” “containing,” or any other variations thereof, as used herein, are intended to cover non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0017] The conjunction "composed of..." excludes any unspecified elements, steps, or components. If used in a claim, this phrase makes the claim closed, excluding materials other than those described, except for associated conventional impurities. When the phrase "composed of..." appears in a clause of the body of a claim rather than immediately following it, it limits only the elements described in that clause; other elements are not excluded from the claim as a whole.

[0018] When a quantity, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pair of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range “1 to 5” is disclosed, the described range should be interpreted as including the ranges “1 to 4”, “1 to 3”, “1 to 2”, “1 to 2 and 4 to 5”, “1 to 3 and 5”, etc. When numerical ranges are described herein, unless otherwise stated, the range is intended to include its endpoints and all integers and fractions within that range.

[0019] The singular form includes the plural objects of discussion unless the context clearly indicates otherwise. "Optional" or "any one" means that the matter or event described thereafter may or may not occur, and the description includes both the possibility that the event occurs and the possibility that the event does not occur.

[0020] Approximate terms used in the specification and claims to modify quantities indicate that the invention is not limited to that specific quantity, but also includes acceptable modifications close to that quantity that do not alter the relevant essential function. Correspondingly, the use of "about," "approximately," etc., to modify a numerical value means that the invention is not limited to that precise value. In some instances, approximate terms may correspond to the precision of the instrument used to measure the value. In this application's specification and claims, scope definitions can be combined and / or interchanged, unless otherwise stated, these scopes include all subscopes contained therein.

[0021] Furthermore, the indefinite articles “a” and “an” preceding the elements or components of this invention do not impose any limitation on the quantity (i.e., number of times) of the elements or components. Therefore, “an” or “a” should be interpreted as including one or at least one, and the singular form of an element or component also includes the plural form, unless the quantity clearly refers to the singular form.

[0022] Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention. The preferred embodiments and materials described herein are for illustrative purposes only and do not limit the scope of this application.

[0023] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; unless otherwise specified, the experimental materials and test strains used in the following examples were purchased from commercial channels.

[0024] Raw material standards and pretreatment instructions: The core matrix used in this invention, "sweet potato vinegar base," refers to fermented, mature vinegar made primarily from sweet potatoes through a complete biological fermentation process, including saccharification, alcoholic fermentation, and acetic acid fermentation, without blending, dilution, or filtration. It provides the acidic extraction environment, specific flavor, and some active ingredients for the composition of this invention.

[0025] To ensure the stability of the extraction process and the consistency of the final product quality, the sweet potato vinegar concentrate should meet the following technical specifications: Total acid content (calculated as acetic acid): should be 4.0-6.0 g / 100mL. This acidity range can effectively extract alkaloids, flavonoids and other substances from herbal ingredients, while ensuring that the pH value of the final product is compatible with the physiological environment of the nasal mucosa (slightly acidic) and avoids excessive irritation.

[0026] pH value: should be 3.0-3.8. This pH range is one of the indicators of mature acetic acid fermentation and also helps to inhibit the growth of some microorganisms during the process.

[0027] Soluble solids: not less than 5.0°Brix. Adequate solids (including undigested sugars, amino acids, minerals, etc.) contribute to a more stable system during extraction and homogenization.

[0028] Before use, the sweet potato vinegar concentrate should be preliminarily tested, and it can only be put into production if it meets the above indicators. If there are differences in total acid content between batches, the process can be fine-tuned by adjusting the extraction temperature or time through small-scale trials, but the core indicators must be within the above-mentioned range.

[0029] Explanation of the functions of key excipients: The ultrafine charcoal powder deliberately added in this invention is not merely for forming a physically stable suspension system, but also enhances the composition's ability to regulate the overall nasal environment through multiple synergistic mechanisms, directly related to its core efficacy of relieving dryness, tightness, and poor ventilation. Adsorption and purification mechanism indirectly relieves mucosal irritation and dryness: Ultrafine charcoal has a huge specific surface area and abundant microporous structure, which can efficiently adsorb abnormal metabolites, trace amounts of inflammatory mediators (such as histamine and leukotrienes) in nasal secretions, as well as allergens or dust particles that may be inhaled from the environment. Through this local micro-purification effect, the continuous stimulation of these substances on the nerve endings of the nasal mucosa is reduced, thereby helping to relieve reflexive mucosal congestion, thick secretions, and subjective dryness, itching, and foreign body sensation caused by irritation.

[0030] Assisting in the regulation of the nasal microenvironment and supporting the relief of nasal congestion and tightness: As shown in Experimental Example 5 of this invention, the charcoal component has a significant inhibitory effect on biofilm formation of opportunistic pathogens such as Staphylococcus aureus and Candida albicans. Biofilms are protective communities of bacteria against external environments and drugs, and their formation is closely related to chronic nasal inflammation, persistent abnormal secretions, and mucosal swelling. By interfering with biofilm formation, charcoal helps break the vicious cycle of the nasal microecology, reducing mucosal swelling and secretion accumulation caused by excessive microbial proliferation or the physical barrier of biofilms, thereby indirectly promoting nasal ventilation and relieving mucosal tightness and discomfort caused by obstruction and inflammation.

[0031] Carrier and sustained-release potential enhance efficacy: After the charcoal powder is uniformly dispersed in the system, its porous structure can adsorb some of the herbal active ingredients and flavor molecules. During use, these adsorbed components may be slowly released over time, thus exerting a certain sustained-release effect locally, which helps to prolong the sensory action time and efficacy duration of the product, enhancing the user's continuous comfort experience.

[0032] Stable System and Aesthetic Benefits: The processed ultrafine charcoal powder is homogenized and dispersed in the system at high speed, effectively forming a fine and uniform suspension. This prevents oil phase aggregation and solid sedimentation, ensuring the physical stability of the product throughout its shelf life and the uniformity of dosage for each use. Simultaneously, it gives the product an elegant light gray appearance, enhancing its professional image.

[0033] Process design principle explanation: The preparation method of this invention employs a sequential extraction strategy combining stepwise dynamic countercurrent extraction (corresponding to step S2) with integrated extraction of flavor and heat-sensitive components (corresponding to step S3). This design, based on the physicochemical properties of different raw materials and the differences in the thermal stability of target components, aims to achieve efficient, targeted enrichment and maximum protection of all active ingredients. The specific principle is as follows: The purpose and positioning of stepwise dynamic countercurrent extraction (S2): This step mainly targets dried herbal raw materials with dense texture and strong cell wall structure, such as Xanthium sibiricum, Magnolia biondii, Angelica dahurica, and Sophora flavescens. The target components contained in these materials, such as sesquiterpene lactones (e.g., xanthogenoside), volatile oils (e.g., angelicin), alkaloids, and flavonoid glycosides, have a slow dissolution rate, requiring a certain thermodynamic driving force and duration to ensure sufficient extraction. Using sweet potato vinegar as a solvent, the plant cell walls can be initially broken down under ultrasonic assistance (50±2℃). Then, a high concentration gradient within the system is maintained through dynamic countercurrent circulation (68±2℃) to continuously promote component diffusion. The controlled upper temperature limit (≤70℃) and slightly negative pressure environment in this stage aim to prevent the hydrolysis or oxidation of dissolved components in the vinegar due to prolonged high temperatures, while ensuring deep extraction of the core components of the herbal powder.

[0034] Positioning and Purpose of Integrated Extraction of Flavor and Heat-Sensitive Components (S3): This step primarily targets tender plant tissues such as perilla leaves, ginger, and scallion whites, as well as raw materials rich in volatile and heat-sensitive active substances, such as garlic and Sichuan pepper. Key components in these materials (such as perillaldehyde, gingerol, allicin, and safflower extract) have small molecular weights, low boiling points, and are sensitive to heat and oxygen. If extracted together with herbal powders at a relatively high temperature for an extended period in stage S2, it can easily lead to significant volatilization losses, chemical structure damage or transformation, thereby significantly reducing the flavor intensity and some biological activity of the product. Therefore, after S2, the primary alcohol extract is cooled and then subjected to a closed-loop warm maceration under mild conditions (60±2℃). At this point, the primary alcohol extract becomes a carrier rich in acetic acid-soluble components. Its acidic environment and existing components promote the dissolution of target components in fresh materials and spices, while the relatively low temperature and closed system effectively lock in volatile substances and protect the integrity of heat-sensitive components.

[0035] The combined advantages of the step-by-step process: Through the sequential operation of "first deeply extracting tough herbs, then gently extracting fresh and tender spices," the following is achieved: Extraction selectivity optimization: Based on the physicochemical properties of different raw materials, avoid component loss or damage caused by "one-size-fits-all" extraction conditions.

[0036] Maximizing the retention of active ingredients: While ensuring the extraction rate of traditional herbal medicinal ingredients, it greatly increases the final retention of volatile substances and heat-sensitive active ingredients in the product.

[0037] Example 1 A herbal composition for nasal care, comprising, by weight fraction, the following ingredients: 800 parts sweet potato vinegar, 12 parts cocklebur, 6 parts magnolia flower, 4 parts angelica root, 3 parts perilla leaf, 25 parts garlic, 4 parts Sichuan pepper, 4 parts sesame oil, 5 parts honey, 2 parts ginger, 2 parts scallion, 4 parts sophora flavescens, and 0.8 parts charcoal.

[0038] A method for preparing an herbal composition for nasal care includes the following steps: S1. Raw material pretreatment: Xanthium sibiricum, Magnolia biondii, Angelica dahurica, and Sophora flavescens are cleaned, microwave-dried to a moisture content of ≤8%, then pulverized using a universal pulverizer and passed through an 80-mesh sieve to obtain herbal powder; Perilla frutescens leaves, ginger, and scallion whites are washed, sterilized with ozone water, and then cut into 4mm segments to obtain fresh materials for use; Garlic is peeled and washed, and then pulverized with Sichuan peppercorns using a low-temperature airflow pulverizer to a particle size D90≤150μm to obtain garlic micro powder and Sichuan peppercorn micro powder; Charcoal is ultra-finely pulverized under inert gas protection and passed through an 800-mesh sieve to obtain ultra-fine charcoal powder, which is then placed in a moisture-proof container for later use; S2. Stepwise dynamic countercurrent extraction: The herbal powder obtained in step S1 and all the sweet potato vinegar pulp were put into a multi-functional hot reflux extraction tank. Stirring was turned on, and the temperature was raised to 48°C at a rate of 1.5°C / min. The temperature was maintained and ultrasonically assisted extraction was performed at a power density of 50W / L for 45min. The temperature was then raised to 66°C at a rate of 1°C / min. Dynamic countercurrent circulation extraction was performed at this temperature for 4 hours. During the extraction process, a slight negative pressure of -0.03MPa was maintained inside the tank. After the extraction was completed, the liquid was cooled to below 40°C through the built-in cooling coil to obtain the primary alcohol extract. Specifically, dynamic countercurrent circulation extraction involves pumping the liquid from the bottom of the extraction tank to a spray device on the top of the tank to form a continuous internal circulation with a circulation flow rate of twice the tank volume per hour.

[0039] S3. Integrated extraction of flavor and heat-sensitive components: The fresh material to be used, garlic powder and Sichuan pepper powder obtained in step S1 are added together to the primary alcohol extract and warm-soaked for 90 minutes at 58°C and under sealed conditions with a stirring speed of 120 rpm. S4. Precision separation and purification: The mixture after step S3 is fed into a tubular centrifuge and solid-liquid separation is performed under a centrifugal force of 10000G. The supernatant is collected. The supernatant is then filtered sequentially through a 50μm bag filter and a 5μm precision filter to obtain a clear extract. Specifically, after the tubular centrifugation, the obtained solid residue is rinsed with 30% of its mass of purified water. The rinsing liquid is collected separately, concentrated at low temperature, and then added back to the supernatant to improve the transfer rate of the target component.

[0040] S5. Controllable homogenization and stabilization: The clarified extract is introduced into a jacketed cooling mixing tank, cooled and maintained at 22°C; under continuous stirring at 80 rpm / min, sesame oil and honey are slowly and evenly added to the tank, and stirred for 20 min until completely emulsified and dispersed; with the high-speed shear emulsifier running at 10000 rpm / min, the ultrafine charcoal powder obtained in step S1 is evenly drawn into the emulsifier head through a vacuum feeding device, and homogenized continuously for 15 min to form a stable suspension system; Specifically, the sesame oil is subjected to nitrogen bubbling deoxygenation treatment for 10 minutes before use, and the honey is filtered through a 40-mesh sieve to remove crystallized particles and impurities.

[0041] Meanwhile, before being inhaled, the ultrafine charcoal powder is premixed with an equal weight of clarified extract to form a slurry, which is then quantitatively delivered to the emulsifying head via a peristaltic pump.

[0042] S6. Aseptic filling: The homogenized semi-finished product is filtered through a 0.45μm terminal sterilization filter and then transferred to an aseptic temporary storage tank; using a light-proof and well-sealed pharmaceutical nasal drop bottle or spray bottle, the product is filled and capped in a Class B clean environment to obtain the finished herbal composition for nasal care.

[0043] Specifically, before filling, high-purity nitrogen is introduced into the sterile temporary storage tank to replace the air at the top of the tank for 10 minutes, and the tank is kept at a slightly positive nitrogen pressure of 0.05 MPa until filling is completed.

[0044] Example 2 A herbal composition for nasal care, comprising the following ingredients by weight fraction: 1000 parts sweet potato vinegar, 15 parts cocklebur, 8 parts magnolia flower, 6 parts angelica, 4 parts perilla leaf, 30 parts garlic, 5 parts Sichuan pepper, 5 parts sesame oil, 6 parts honey, 3 parts ginger, 3 parts scallion, 5 parts sophora flavescens, and 1 part charcoal.

[0045] A method for preparing an herbal composition for nasal care includes the following steps: S1. Raw material pretreatment: Xanthium sibiricum, Magnolia biondii, Angelica dahurica, and Sophora flavescens are cleaned, microwave-dried to a moisture content of ≤8%, then pulverized using a universal pulverizer and passed through an 80-mesh sieve to obtain herbal powder; Perilla frutescens leaves, ginger, and scallion whites are washed, sterilized with ozone water, and then cut into 5mm segments to obtain fresh materials for use; Garlic is peeled and washed, and then pulverized with Sichuan peppercorns using a low-temperature airflow pulverizer to a particle size D90≤150μm to obtain garlic micro powder and Sichuan peppercorn micro powder; Charcoal is ultra-finely pulverized under inert gas protection and passed through an 800-mesh sieve to obtain ultra-fine charcoal powder, which is then placed in a moisture-proof container for later use; S2. Stepwise dynamic countercurrent extraction: The herbal powder obtained in step S1 and all the sweet potato vinegar pulp were put into a multi-functional hot reflux extraction tank. Stirring was turned on, and the temperature was raised to 50℃ at a rate of 1.5℃ / min. The temperature was maintained and ultrasonically assisted extraction was performed at a power density of 50W / L for 45min. The temperature was then raised to 68℃ at a rate of 1℃ / min. Dynamic countercurrent circulation extraction was performed at this temperature for 4 hours. During the extraction process, a slight negative pressure of -0.025MPa was maintained inside the tank. After the extraction was completed, the liquid was cooled to below 40℃ through the built-in cooling coil to obtain the primary alcohol extract. Specifically, dynamic countercurrent circulation extraction involves pumping the liquid from the bottom of the extraction tank to a spray device on the top of the tank, forming a continuous internal circulation with a circulation flow rate of 2.5 times the tank volume per hour.

[0046] S3. Integrated extraction of flavor and heat-sensitive components: The fresh material to be used, garlic powder and Sichuan pepper powder obtained in step S1 are added together to the primary alcohol extract and warm-soaked for 90 minutes at 60°C and under sealed conditions with a stirring speed of 120 rpm. S4. Precision separation and purification: The mixture after step S3 is fed into a tubular centrifuge and solid-liquid separation is performed under a centrifugal force of 10000G. The supernatant is collected. The supernatant is then filtered sequentially through a 50μm bag filter and a 5μm precision filter to obtain a clear extract. Specifically, after the tubular centrifugation, the obtained solid residue is rinsed with 30% of its mass of purified water. The rinsing liquid is collected separately, concentrated at low temperature, and then added back to the supernatant to improve the transfer rate of the target component.

[0047] S5. Controllable homogenization and stabilization: The clarified extract is introduced into a jacketed cooling mixing tank, cooled and maintained at 25°C; under continuous stirring at 80 rpm / min, sesame oil and honey are slowly and evenly added to the tank, and stirred for 20 min until completely emulsified and dispersed; with the high-speed shear emulsifier running at 10000 rpm / min, the ultrafine charcoal powder obtained in step S1 is evenly drawn into the emulsifier head through a vacuum feeding device, and homogenized continuously for 15 min to form a stable suspension system; Specifically, the sesame oil is subjected to nitrogen bubbling deoxygenation treatment for 10 minutes before use, and the honey is filtered through a 40-mesh sieve to remove crystallized particles and impurities.

[0048] Meanwhile, before being inhaled, the ultrafine charcoal powder is premixed with an equal weight of clarified extract to form a slurry, which is then quantitatively delivered to the emulsifying head via a peristaltic pump.

[0049] S6. Aseptic filling: The homogenized semi-finished product is filtered through a 0.45μm terminal sterilization filter and then transferred to an aseptic temporary storage tank; using a light-proof and well-sealed pharmaceutical nasal drop bottle or spray bottle, the product is filled and capped in a Class B clean environment to obtain the finished herbal composition for nasal care.

[0050] Specifically, before filling, high-purity nitrogen is introduced into the sterile temporary storage tank to replace the air at the top of the tank for 10 minutes, and the tank is kept at a slightly positive nitrogen pressure of 0.04 MPa until filling is completed.

[0051] Example 3 A herbal composition for nasal care, comprising the following ingredients by weight fraction: 1200 parts sweet potato vinegar, 18 parts cocklebur, 10 parts magnolia flower, 8 parts angelica, 5 parts perilla leaf, 35 parts garlic, 6 parts Sichuan pepper, 6 parts sesame oil, 7 parts honey, 4 parts ginger, 4 parts scallion, 6 parts sophora flavescens, and 1.2 parts charcoal.

[0052] A method for preparing an herbal composition for nasal care includes the following steps: S1. Raw material pretreatment: Xanthium sibiricum, Magnolia biondii, Angelica dahurica, and Sophora flavescens are cleaned, microwave-dried to a moisture content of ≤8%, then pulverized using a universal pulverizer and passed through an 80-mesh sieve to obtain herbal powder; Perilla frutescens leaves, ginger, and scallion whites are washed, sterilized with ozone water, and then cut into 6mm segments to obtain fresh materials for use; Garlic is peeled and washed, and then pulverized with Sichuan peppercorns using a low-temperature airflow pulverizer to a particle size D90≤150μm to obtain garlic micro powder and Sichuan peppercorn micro powder; Charcoal is ultra-finely pulverized under inert gas protection and passed through an 800-mesh sieve to obtain ultra-fine charcoal powder, which is then placed in a moisture-proof container for later use; S2. Stepwise dynamic countercurrent extraction: The herbal powder obtained in step S1 and all the sweet potato vinegar pulp were put into a multi-functional hot reflux extraction tank. Stirring was turned on, and the temperature was raised to 52°C at a rate of 1.5°C / min. The temperature was maintained and ultrasonically assisted extraction was performed at a power density of 50W / L for 45min. The temperature was then raised to 70°C at a rate of 1°C / min. Dynamic countercurrent circulation extraction was performed at this temperature for 4 hours. During the extraction process, a slight negative pressure of -0.02MPa was maintained inside the tank. After the extraction was completed, the liquid was cooled to below 40°C through the built-in cooling coil to obtain the primary alcohol extract. Specifically, dynamic countercurrent circulation extraction involves pumping the liquid from the bottom of the extraction tank to a spray device on the top of the tank to form a continuous internal circulation, with a circulation flow rate of 2-3 times the tank volume per hour.

[0053] S3. Integrated extraction of flavor and heat-sensitive components: The fresh material to be used, garlic powder and Sichuan pepper powder obtained in step S1 are added together to the primary alcohol extract and warm-soaked for 90 min at 60±2℃ and under sealed conditions with a stirring speed of 120 rpm. S4. Precision separation and purification: The mixture after step S3 is fed into a tubular centrifuge and solid-liquid separation is performed under a centrifugal force of 10000G. The supernatant is collected. The supernatant is then filtered sequentially through a 50μm bag filter and a 5μm precision filter to obtain a clear extract. Specifically, after the tubular centrifugation, the obtained solid residue is rinsed with 30% of its mass of purified water. The rinsing liquid is collected separately, concentrated at low temperature, and then added back to the supernatant to improve the transfer rate of the target component.

[0054] S5. Controllable homogenization and stabilization: The clarified extract is introduced into a jacketed cooling mixing tank, cooled and maintained at 28°C; under continuous stirring at 80 rpm / min, sesame oil and honey are slowly and evenly added to the tank, and stirred for 20 min until completely emulsified and dispersed; with the high-speed shear emulsifier running at 10000 rpm / min, the ultrafine charcoal powder obtained in step S1 is evenly drawn into the emulsifier head through a vacuum feeding device, and homogenized continuously for 15 min to form a stable suspension system; Specifically, the sesame oil is subjected to nitrogen bubbling deoxygenation treatment for 10 minutes before use, and the honey is filtered through a 40-mesh sieve to remove crystallized particles and impurities.

[0055] Meanwhile, before being inhaled, the ultrafine charcoal powder is premixed with an equal weight of clarified extract to form a slurry, which is then quantitatively delivered to the emulsifying head via a peristaltic pump.

[0056] S6. Aseptic filling: The homogenized semi-finished product is filtered through a 0.45μm terminal sterilization filter and then transferred to an aseptic temporary storage tank; using a light-proof and well-sealed pharmaceutical nasal drop bottle or spray bottle, the product is filled and capped in a Class B clean environment to obtain the finished herbal composition for nasal care.

[0057] Specifically, before filling, high-purity nitrogen is introduced into the sterile temporary storage tank to replace the air at the top of the tank for 10 minutes, and the tank is kept at a slightly positive nitrogen pressure of 0.03 MPa until filling is completed.

[0058] Experiment Example 1: Human Trial Experience and Subjective Improvement Evaluation Experiment 1. Experimental objective: Human trials were conducted to collect data on the subjective improvement of discomfort such as nasal dryness and tightness in the target population after using the herbal composition of this invention, and to evaluate the comfort and overall acceptance of the product.

[0059] 2. Test materials: Test substance: The herbal composition of the present invention prepared in Example 2 of the present invention (batch number: S230502).

[0060] 3. Experimental Design: Design type: multi-center, single-arm, open-ended, self-controlled experiential study.

[0061] Study period: 14-day product trial period + baseline period + follow-up period.

[0062] 4. Subjects: 4.1 Selection Criteria: 4.1.1 Age 18-65 years old, gender not limited.

[0063] 4.1.2 Self-report that you have experienced nasal dryness and / or tightness in your nasal cavity frequently (≥3 days / week) in the past month, and that this discomfort is not caused by an acute upper respiratory tract infection (cold).

[0064] 4.1.3 Voluntary participation and signing of informed consent form.

[0065] 4.2 Exclusion criteria: 4.2.1 Individuals with known hypersensitivity to any component of this product.

[0066] 4.2.2 Those suffering from acute rhinitis, sinusitis, or active bleeding or ulceration in the nasal cavity.

[0067] 4.2.3 Individuals who have undergone nasal surgery or other intranasal drug treatments within the past month.

[0068] 4.2.4 Pregnant and lactating women.

[0069] 4.2.5 Other situations in which researchers deem it unsuitable for participation (e.g., inability to adhere to the research protocol).

[0070] Sample size: We plan to recruit 120 participants, considering a 10% dropout rate, with the goal of obtaining at least 108 valid data points.

[0071] 5. Experimental Procedure: 5.1 Screening and Enrollment (D-7~D-1): Potential subjects are recruited through advertising, and online / offline screening is conducted. Those who meet the criteria sign an informed consent form.

[0072] 5.2 Baseline Assessment (D0): On the day of enrollment, participants completed a baseline electronic questionnaire, retrospectively rating their nasal discomfort over the past week. Researchers then distributed the product and provided on-site standardized usage training (video + explanation) to ensure each participant mastered the correct cleaning, drop / spray, and massage techniques.

[0073] 5.3 Product trial period (D1-D14): Subjects used the product at home 2-3 times a day as required and kept a usage diary.

[0074] 5.4 Follow-up and evaluation: Day 7 follow-up (D7±1): Researchers followed up by phone or online questionnaire, reminding participants to complete the Day 7 electronic questionnaire and inquiring about compliance and any discomfort.

[0075] Endpoint assessment on day 14 (D14±2): Participants returned to the research center, submitted their product and usage diaries, completed the day 14 electronic questionnaire, and participated in an overall experience interview.

[0076] 6. Evaluation Indicators and Methods: All questionnaires were completed using a controlled electronic data collection system.

[0077] Key evaluation indicators: changes in visual analog scale (VAS) scores for nasal dryness, nasal tightness / discomfort, and subjective perception of nasal ventilation.

[0078] Methods: A 0-10 VAS scale was used. 0 points represented "no dryness / tightness / blockage at all," and 10 points represented "extreme dryness / tightness / blockage, unbearable." Participants dragged a slider on an electronic device to rate their scores.

[0079] Secondary evaluation indicators: 1. Product comfort rating (VAS): 0 points represents "very irritating and uncomfortable", and 10 points represents "very mild and comfortable".

[0080] 2. Overall Satisfaction VAS Rating: 0 points represents "very dissatisfied", and 10 points represents "very satisfied".

[0081] 3. Product compliance: Calculate the percentage of actual usage times compared to planned usage times using a diary.

[0082] 4. Adverse event record: All adverse reactions reported by the subject through proactive inquiry and voluntary reporting.

[0083] 7. Experimental Data Table 1: Changes in VAS scores of key evaluation indicators ( Valid samples )

[0084] Note: Nasal airflow perception score. The higher the score, the stronger the feeling of blockage and the worse the airflow.

[0085] Table 2: Results of Secondary Evaluation Indicators ( (or percentage)

[0086] Note: The main adverse events were 3 people who reported a slight cooling sensation upon first use and 2 people who reported a brief (<1 minute) mild nasal itching. All of these events resolved on their own and did not lead to withdrawal.

[0087] 8. Experimental Conclusion: 8.1 The product can quickly and significantly improve the discomfort of the target population: As shown in Table 1, after only 7 days of product use, the VAS scores for "nasal dryness," "tightness," and "ventilation" (congestion) showed a highly significant decrease compared to baseline (P<0.001), with an average decrease of over 50%. This demonstrates that the product can effectively alleviate the main symptoms affecting the target population.

[0088] By day 14, the improvement in the three discomfort scores had further deepened, with the average score dropping below 2.3 (the "mild" range). This indicates that the product's improvement is not temporary, but rather brings a better and more stable comfort experience with continued use.

[0089] 8.2 The product exhibits excellent gentleness and user acceptance: Table 2 shows that the product's comfort score reached as high as 8.5 and 8.9 on day 7 and day 14, respectively, and the overall satisfaction score also significantly increased from 7.8 to 8.6. These high scores indicate that the vast majority of participants found the product gentle and non-irritating, resulting in a positive user experience.

[0090] The extremely high average compliance rate (>92%) behaviorally confirms the subjects' acceptance and recognition of the product. The low incidence of adverse events (4.5%) and the mild and transient symptoms further confirm the product's local safety, consistent with the high comfort score.

[0091] Experimental Example 2: Local Tolerance and Human Skin Patch Test 1. Experimental objective: The herbal composition of this invention was evaluated directly on healthy human skin using an internationally recognized closed patch test, providing direct evidence of its safety for topical use.

[0092] 2. Experimental Design: Design type: Single-center, randomized, assessor-blinded controlled trial.

[0093] In accordance with the standards: The test protocol is mainly based on the "Human Skin Patch Test" method in the "Cosmetic Safety Technical Specifications" and refers to the relevant guidelines of ICH (International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use).

[0094] 3. Subjects: 3.1 Selection Criteria: 3.1.1 Healthy volunteers aged 18-60 years, half male and half female.

[0095] 3.1.2 The skin on the flexor side of the upper forearm is healthy and intact, without dermatitis, eczema, pigmentation, scars, etc.

[0096] 3.1.3 No known history of allergies, especially to plant extracts, fragrances, honey, etc.

[0097] 3.1.4 Voluntary participation and signing of a written informed consent form.

[0098] 3.2 Exclusion criteria: 3.2.1 Women who plan to become pregnant during pregnancy, breastfeeding, or during a trial period.

[0099] 3.2.3 Individuals who have used topical medications or functional skincare products on their upper limbs or forearms within the past week.

[0100] 3.2.3 Individuals with a history of severe systemic diseases, immune diseases, or skin diseases.

[0101] 3.2.4 Other situations that researchers deemed unsuitable for participation.

[0102] Sample size: We plan to recruit 32 volunteers, taking into account the possibility of dropout, to ensure at least 30 valid data.

[0103] 4. Test materials: Test substance: The herbal composition of the present invention (batch number: S230502) prepared in Example 2 of the present invention, the undiluted solution was used directly.

[0104] Control: Negative control: 0.9% sterile saline.

[0105] Positive control: 1% sodium dodecyl sulfate aqueous solution.

[0106] Patching equipment: standard FinnChamber (8mm diameter) patch applicator and hypoallergenic porous medical tape.

[0107] 5. Experimental Procedure: 5.1 Preparation and application: Volunteers were randomly numbered, and test areas were determined on the flexor surfaces of the left and right forearms (at least 4 cm apart).

[0108] Take approximately 15 μL of the test substance and the negative control, and add them separately to different Finn Chambers.

[0109] The loaded spot tester was symmetrically applied to the healthy skin on the flexor side of one forearm of the volunteer (the test substance and negative control were randomly assigned to the left and right positions) and fixed with tape.

[0110] Volunteers were instructed to keep the patch area dry and avoid friction, pressure, and direct sunlight.

[0111] 5.2 Closed contact: The patch duration is 24 hours.

[0112] 5.3 Removal and observation: After 24 hours, the spot device was carefully removed by a trained researcher (blinded by the test substance number).

[0113] Initial observation (30 minutes after removal): After the temporary pressure marks caused by the adhesive have faded, perform the first skin reaction assessment.

[0114] Follow-up observation: A second and third follow-up observation were conducted 24 hours and 48 hours after the patch was removed.

[0115] At each observation, gently wipe the test area with an alcohol swab to remove residue, and wait 1 minute before evaluating.

[0116] 5.4 Skin Reaction Score: Two independent assessors conducted a double-blind scoring process based on the skin adverse reaction grading criteria in the "Cosmetic Safety Technical Specifications" (see table below). The highest reaction score for each subject at each time point was recorded.

[0117] Skin reaction scoring criteria:

[0118] 6. Experimental Data: Table 1: Incidence and mean score of skin patch test reaction (valid sample n=30)

[0119] Note: Response rate = (Number of participants with a score ≥1 / Total number of participants) × 100%. The average response score was calculated from the scores of all 30 participants.

[0120] Table 2: Specific Skin Reactions of Test Substance Groups

[0121] System validation (preliminary experiment): In a parallel positive control group (1% SDS) of 5 volunteers, all showed a clear erythema reaction with a score ≥2 (mean score 2.4±0.5) 24 hours after removal, confirming that the detection system has the sensitivity to identify irritants.

[0122] 7. Experimental Conclusion: 7.1 This product exhibits an extremely low reaction rate to human skin, and the reaction is mild and transient.

[0123] As shown in Table 1, among the 30 subjects, only 1 person (3.3%) experienced a “mild erythema” with a score of 1 30 minutes after removal and 2 people (6.7%) experienced a “mild erythema” 24 hours after removal. All reactions completely subsided within 48 hours, with the average reaction score dropping to 0. This indicates that even if a reaction occurred, its intensity was only “barely visible” and self-limiting.

[0124] 7.2 The skin reaction characteristics of this product were not statistically different from those of the negative control (physiological saline).

[0125] Although a few mild reactions occurred in the test substance group, the average reaction score was extremely low at all time points (0.00-0.07), and there was no significant difference in statistical analysis (e.g., Mann-Whitney U test) compared to the 0 score in the saline group (P>0.05). Based on the population data, the skin reaction profile of this product is similar to that of non-irritating saline.

[0126] 7.3 Tests have confirmed that this product meets the safety requirements for external use products.

[0127] According to the "Cosmetic Safety Technical Specifications," skin patch test results are generally interpreted as follows: a reaction rate of <10% indicates that the product has very little or no irritation. In this test, the highest reaction rate was 6.7%, which is below this threshold.

[0128] Based on the fact that all reactions are the mildest grade 1 and can be quickly and completely resolved, it can be scientifically determined that the herbal composition of the present invention, under closed conditions, does not produce clinically significant irritation to healthy human skin and has high safety for topical application.

[0129] Experimental Example 3: Verification of the advantages of the preparation process and the stability of the components 1. Experimental objective: This invention verifies the advantages of the integrated preparation process in retaining the characteristic beneficial components of raw materials and maintaining the stability of key quality attributes of the product, proving that the process can produce nasal care products with better intrinsic quality and more stable shelf life.

[0130] 2. Experimental Design: Part 1: Comparative Study of Processes—Comparing the impact of the process of this invention and traditional processes on key product quality indicators.

[0131] Part Two: Accelerated Stability Study—Evaluating the quality stability of the product under accelerated storage conditions.

[0132] 3. Experimental Materials and Methods: 3.1 Sample Preparation Group A (Integrated Process Group of the Invention): Three batches of samples were prepared according to Example 2.

[0133] Group B (Traditional Static Soaking Process Group): Simulates common simple workshop-style production.

[0134] 1. Weigh out all the herbal raw materials in the same amount as Group A.

[0135] 2. Soak the raw materials directly in an equal amount of sweet potato vinegar syrup, seal, and let them soak statically at room temperature (25±2°C) in the dark for 15 days, shaking once a day.

[0136] 3. After soaking, filter with gauze, squeeze the filter residue, and combine the filtrates.

[0137] 4. Add sesame oil and honey directly to the filtrate and mix manually. Do not filter or homogenize at high speed.

[0138] 5. This process simulates traditional methods that lack temperature and time control, precise separation, and homogenization.

[0139] 3.2 Detection Indicators and Methods 3.2.1 Content of characteristic marker components (HPLC method): Perillaldehyde (derived from perilla leaves): Represents the product's characteristic flavor and potential soothing activity. C18 column, acetonitrile-water gradient elution, detection wavelength 230nm.

[0140] 6-Shogaol (from ginger): Represents the product's mild warming sensation and auxiliary activity. C18 column, methanol-water gradient elution, detection wavelength 280nm.

[0141] Total flavonoid content: Represents the product's overall antioxidant and auxiliary potential. Measured at 510 nm using the aluminum nitrate colorimetric method, expressed as rutin.

[0142] 3.2.2 Key auxiliary material quality indicators: Honey amylase value: reflects whether honey has undergone improper high-temperature processing and represents the degree of retention of natural activity. It is determined according to the method specified in GB14963-2011 National Food Safety Standard for Honey.

[0143] Peroxide value of sesame oil: reflects the degree of initial oxidation of the oil and indicates the product's oxidative stability. It is determined according to the method specified in GB5009.227-2023 National Food Safety Standard for Determination of Peroxide Value in Food.

[0144] 3.2.3 Accelerated stability test Conditions: Place the three batches of finished products in Group A in a constant temperature and humidity chamber at 40°C±2°C and 75%±5% relative humidity.

[0145] Sampling points: 0 months (initial), 1 month, 2 months, 3 months, 6 months.

[0146] Testing indicators: appearance / uniformity, pH value, perillaldehyde content, total flavonoid content, peroxide value, and microbial limits.

[0147] 4. Experimental data: Table 1: Results of the process comparison study ( )

[0148] Note: The lower amylase value in group B may be due to partial enzyme inactivation caused by prolonged soaking at room temperature; the higher peroxide value indicates that the oil is more easily oxidized without protection. RE is rutin equivalent.

[0149] Table 2: Changes in key indicators of accelerated stability test of finished products in Group A (process of this invention)

[0150] Note: Retention rate is based on 100% of data from the 0-month period. Peroxide value increases slowly, but remains well below the common safety limit of 0.10g / 100g at 6 months.

[0151] 5. Experimental Conclusion: 5.1 The process of this invention can significantly improve product quality and more completely preserve the essence of the raw materials: As shown in Table 1, compared with the simulated traditional static process (Group B), the product produced using the process of this invention (Group A) has more than 150% higher content of characteristic marker components (perillaldehyde, 6-shogaol) and total flavonoids. This directly demonstrates the advantages of the "stepwise dynamic countercurrent extraction" and "flavor integration extraction" processes in terms of efficient and targeted enrichment of beneficial components, ensuring that the final product contains richer active substances from the raw materials.

[0152] Meanwhile, the honey in Group A products had a higher amylase value and the sesame oil had a lower peroxide value, indicating that this process, through precise low-temperature control and anaerobic / oxygen-free operation, better protects the natural quality and freshness of heat-sensitive and easily oxidized auxiliary materials.

[0153] 5.2 The process of this invention is the fundamental guarantee for the product to obtain excellent physical stability: The contrast in product appearance in Table 1 is striking: Group A is a "homogeneous and fine suspension," while Group B shows "obvious stratification." This demonstrates that the "controlled homogenization and stabilization" process can fuse various components with different properties (aqueous phase, oil phase, and solid particles) into a homogeneous, stable consumer product suitable for shelf life.

[0154] 5.3 Accelerated stability testing demonstrates that the product of this invention possesses excellent shelf-life stability: Table 2 shows that under accelerated conditions (6 months, 40°C / 75%RH) equivalent to about 2 years of storage at room temperature, the product's physical state remained stable with no demulsification or stratification; the retention rates of characteristic components (perillaldehyde and total flavonoids) were all above 90%, indicating that its chemical properties were stable; the peroxide value increased slowly and remained at a safe low level, indicating that the antioxidant system was effective; and no microorganisms were detected, proving that the aseptic filling process was reliable.

[0155] Test Example 4: Evaluation of Product Mildness and Nasal Mucosal Cell Safety (In Vitro) 1. Experimental objective: Using a human nasal epithelial cell model, the direct effects of the herbal composition of this invention on nasal mucosal cells were evaluated in vitro, and its cytotoxicity and potential impact on inflammatory responses were detected. This provides a scientific basis from a cell biology perspective for the product's mildness, non-irritation, and safety for external use.

[0156] 2. Test materials: Cell line: Human nasal epithelial cell line RPMI2650 (ATCCCCL-30). This cell line is widely used to simulate the human nasal epithelial barrier and for drug / irritant toxicity studies.

[0157] Test substance: The herbal composition prepared in Example 2 of this invention (batch number: S230502). Before use, it was sterile filtered (0.22 μm filter membrane) with cell-specific culture medium and serially diluted.

[0158] Reference standard: Blank control: Complete culture medium.

[0159] Negative control: Complete culture medium containing 0.1% DMSO (used to verify that the solvent has no effect).

[0160] Positive control (cytotoxicity): Complete culture medium containing 1% Triton X-100 (can cause 100% cell death).

[0161] Positive control (inflammation induction): Complete culture medium containing 10 ng / mL tumor necrosis factor-α (TNF-α).

[0162] Main reagents and instruments: RPMI 1640 medium, fetal bovine serum, CCK-8 kit, human IL-8 and IL-1β ELISA kit, CO2 incubator, microplate reader, inverted microscope.

[0163] 3. Experimental Procedure: 3.1 Cell Culture and Seeding: RPMI 2650 cells were cultured routinely in RPMI 1640 medium containing 10% fetal bovine serum at 37°C and 5% CO2. Logarithmic growth phase cells were harvested, digested, and then cultured at 1 × 10⁶ cells per well. 4 Cells were seeded at a density of 1,000 cells per well in 96-well plates and cultured for 24 hours to allow the cells to adhere and enter the logarithmic growth phase.

[0164] 3.2 Test substance treatment and grouping: Cells were randomly divided into the following treatment groups: A. Blank control group: Only fresh complete culture medium was used.

[0165] B. Negative control group: Culture medium containing 0.1% DMSO.

[0166] C. Product Processing Groups (Different Concentrations): The product concentrate was diluted with complete culture medium to four concentrations: 1% (v / v), 5% (v / v), 10% (v / v), and 25% (v / v). This concentration range covers and far exceeds the expected local concentration for actual nasal use (estimated to be <1%).

[0167] D. Positive control (cytotoxicity) group: 1% Triton X-100.

[0168] E. Positive control (inflammation) group: 10 ng / mL TNF-α.

[0169] Each group has 6 duplicate holes.

[0170] 3.3 Detection Indicators and Methods 3.3.1 Cell viability assay (CCK-8 assay): Cells were co-cultured with each treatment solution for 24 hours.

[0171] After the culture is complete, add 10 μL of LCK-8 solution to each well and continue incubation for 2 hours.

[0172] The absorbance (OD value) of each well was measured at a wavelength of 450 nm using an ELISA reader.

[0173] Cell viability (%) = (OD value of treatment group - OD value of blank control group) / (OD value of blank control group - OD value of blank control group) × 100%. Cell viability ≥ 70% is generally considered to be non-cytotoxic.

[0174] 3.3.2 Detection of inflammatory factor release (ELISA method): Another group of cells was cultured for 24 hours with the same grouping and treatment.

[0175] After the culture is completed, carefully collect the supernatant culture medium from each well and store it at -80°C.

[0176] The concentrations of these two key pro-inflammatory factors in the supernatant of each group were measured according to the instructions of the human IL-8 and IL-1β ELISA kit.

[0177] 4. Experimental data: Table 1: Effects of different concentrations of product on the viability of human nasal epithelial cells (24 hours). )

[0178] Table 2: Effects of different product concentrations on the release of inflammatory factors from human nasal epithelial cells (24 hours). )

[0179] Note: P<0.05 compared to the blank control group; ^# indicates P<0.001 compared to the blank control group.

[0180] 5. Experimental Conclusion: 5.1 The product exhibits no cytotoxicity to nasal mucosal cells within the actual usage concentration range, demonstrating high biocompatibility: As shown in Table 1, at dilution concentrations of 1%, 5%, and 10%, the cell viability of the product-treated groups was not statistically different from that of the blank control group, and the viability values ​​were close to or exceeded 100%. This indicates that at these concentrations, the product does not affect the normal growth and metabolism of nasal epithelial cells at all, i.e., it has no cytotoxicity.

[0181] Even at extremely high test concentrations of up to 25%, cell viability remained at 86.4%, significantly higher than the 70% non-toxic threshold. Although there was a slight statistical decrease compared to the control group (P<0.05), from a biological perspective, this slight inhibition does not pose a toxicological hazard, and the concentration is far above the actual exposure concentration.

[0182] 5.2 The product does not induce an inflammatory response in nasal mucosal cells, confirming its mild and non-irritating properties.

[0183] As shown in Table 2, within the concentration range of 1%-10%, the levels of IL-8 and IL-1β released by cells in the product-treated group were basically consistent with those in the blank control group, with no significant increase. This indicates that the product did not activate cellular inflammatory signaling pathways and did not induce or aggravate local inflammation.

[0184] Only at a high concentration of 25% did both inflammatory factors show a weak, statistically significant increase. However, their absolute values ​​(IL-8: 104.2 pg / mL) were far lower than the severe inflammatory levels induced by the positive control TNF-α (IL-8: 525.6 pg / mL), indicating that this weak increase was not clinically or biologically significant, further confirming the mildness of the product formulation.

[0185] 5.3 In vitro cell safety is consistent with previous human trial results: The safe concentration window (at least 1%-10%) determined in this study, which is "non-cytotoxic and non-inflammatory," completely covers and is far greater than the expected local concentration (<1%) after the product is absorbed through the nasal mucosa. This provides a cellular-level mechanistic explanation and solid evidence for the high comfort and low adverse event rate reported in the human trial in Example 1, and the non-irritation shown in the human patch test in Example 2.

[0186] Experimental Example 5: Study on the Moderating Potential of Nasal Cavity Microbiota 1. Experimental objective: The effects of the herbal composition of the present invention on the in vitro growth patterns and biofilm formation capabilities of common nasal microorganisms (including symbiotic bacteria and conditionally pathogenic bacteria) were investigated.

[0187] 2. Experimental Design Concept: This study focuses on the role of health supplements in adjuvant regulation, assessing whether the product can gently influence the growth dynamics and community structure tendencies of microorganisms, particularly inhibiting the formation of biofilms associated with discomfort.

[0188] 3. Test materials: Test substance: The herbal composition prepared in Example 2 of this invention (batch number: S230502). The undiluted solution and a series of concentrations diluted with sterile PBS (1 / 2, 1 / 4, 1 / 8, 1 / 16) were used in the test.

[0189] Test strain: Staphylococcus aureus ATCC25923 (opportunistic pathogen, prone to biofilm formation). Staphylococcus epidermidis ATCC14990 (major commensal). Candida albicans ATCC10231 (common fungus, opportunistic pathogen). Culture media: trypsin-soy broth (TSB), Sabouraud broth (SDB), and 96-well polystyrene plates (for biofilm experiments).

[0190] Main reagents and instruments: crystal violet staining solution, enzyme-linked immunosorbent assay (ELISA) reader, biosafety cabinet.

[0191] 4. Test Procedure 4.1 Effects on microbial planktonic growth (modified agar diffusion method): 4.1.1 Adjust the overnight culture (bacteria to 0.5 McFarland turbidity, fungi 1×10⁻⁶) 6 The CFU / mL concentration was evenly spread onto the corresponding agar plate.

[0192] 4.1.2 Place a sterile Oxford cup and add 100 μL of the test solution, 1 / 2 dilution buffer and negative control (PBS).

[0193] 4.1.3 Incubate upright, bacteria at 37°C for 24 hours, and fungi at 30°C for 48 hours.

[0194] 4.1.4 Measure and record the diameter of a clear inhibition zone. This step is used to qualitatively observe the extent to which the product affects the growth and spread of microorganisms on solid surfaces.

[0195] 4.2 Inhibition potential against microbial biofilm formation (96-well plate micro-method): 4.2.1 In a 96-well plate, dilute the test substance to a series of concentrations (stock solution, 1 / 2, 1 / 4, 1 / 8) with TSB or SDB medium.

[0196] 4.2.2 Add an equal volume of bacterial suspension to each well (final concentration: Staphylococcus aureus ~1×10⁻⁶). 6 CFU / mL; Staphylococcus epidermidis ~1×10 6 CFU / mL; Candida albicans ~1×10 5 (CFU / mL). A bacterial culture growth control without the test substance and a blank control containing no bacteria were set up.

[0197] 4.2.3 Static culture (bacteria 37°C 24 hours; fungi 30°C 48 hours) to promote biofilm formation at the bottom of the well.

[0198] 4.2.4 After the culture is completed, carefully aspirate the floating bacterial solution from each well and gently wash the well walls three times with PBS to remove any unattached cells.

[0199] 4.2.5 Fixation and Staining: Fix each well with 99% methanol for 15 minutes, discard the methanol, and air dry. Stain with 0.1% crystal violet solution for 20 minutes.

[0200] 4.2.6 Wash thoroughly with deionized water until no free dye remains, then air dry. Add 33% glacial acetic acid to each well to dissolve the dye bound to the biofilm, and measure the absorbance (OD value) at a wavelength of 570 nm.

[0201] 4.2.7 Biofilm formation inhibition rate (%) = [1 - (OD value of experimental group - OD value of blank group) / (OD value of growth control group - OD value of blank group)] × 100%.

[0202] 5. Experimental Data Table 1: Effect of the product on the growth and diffusion of microorganisms on solid surfaces (Oxford cup method, inhibition zone diameter, mm, n=3)

[0203] Note: The negative control showed no inhibition zone (Oxford cup pore size only). Data indicate that the product can affect the growth and spread of microorganisms on simulated mucosal surfaces.

[0204] Table 2: Inhibition potential of products on microbial biofilm formation ( )

[0205] 6. Experimental Conclusion: 6.1 The product shows the potential to influence microbial growth patterns, and its effects vary among different strains.

[0206] The results in Table 1 show that the product produces a clear inhibition zone, indicating that its components can diffuse in the local environment and affect the growth of microorganisms. Notably, the diameter of the effect zone on Staphylococcus epidermidis (commensal bacteria) is significantly smaller than that on Staphylococcus aureus and Candida albicans (opportunistic pathogens), suggesting that the product may have a certain selective action tendency.

[0207] 6.2 The product demonstrates a clear inhibitory potential on biofilm formation, which is closely related to nasal discomfort.

[0208] The data in Table 2 show that the product has a concentration-dependent inhibitory effect on biofilm formation of the three test strains at a series of dilution concentrations.

[0209] Especially for Staphylococcus aureus and Candida albicans, two microorganisms associated with chronic nasal discomfort or microecological imbalance, the product can achieve an inhibition rate of over 40%-65% even at 1 / 4 or 1 / 2 dilutions, demonstrating a strong ability to interfere with biofilm formation.

[0210] Although biofilm inhibition of Staphylococcus epidermidis exists, its inhibition rate at the same concentration is generally lower than that against pathogenic bacteria, further confirming the relative selectivity of its action.

[0211] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any person skilled in the art can easily conceive of various variations or substitutions within the technical scope disclosed in the present invention, and these should all be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. An herbal composition for nasal care, characterized in that, By weight fraction, it includes the following ingredients: 800-1200 parts sweet potato vinegar base, 12-18 parts cocklebur, 6-10 parts magnolia flower, 4-8 parts angelica, 3-5 parts perilla leaf, 25-35 parts garlic, 4-6 parts Sichuan pepper, 4-6 parts sesame oil, 5-7 parts honey, 2-4 parts ginger, 2-4 parts scallion, 4-6 parts sophora flavescens, and 0.8-1.2 parts charcoal.

2. The herbal composition for nasal care according to claim 1, characterized in that, By weight fraction, it includes the following ingredients: 1000 parts sweet potato vinegar base, 15 parts cocklebur, 8 parts magnolia flower, 6 parts angelica, 4 parts perilla leaf, 30 parts garlic, 5 parts Sichuan pepper, 5 parts sesame oil, 6 parts honey, 3 parts ginger, 3 parts scallion, 5 parts sophora flavescens, and 1 part charcoal.

3. A method for preparing an herbal composition for nasal care according to claims 1-2, characterized in that, Includes the following steps: S1. Raw material pretreatment: Xanthium sibiricum, Magnolia biondii, Angelica dahurica, and Sophora flavescens are cleaned, microwave-dried to a moisture content of ≤8%, then pulverized using a universal pulverizer and passed through an 80-mesh sieve to obtain herbal powder; Perilla frutescens leaves, ginger, and scallion whites are washed, sterilized with ozone water, and then cut into 5±1mm segments to obtain fresh materials for use; Garlic is peeled and washed, and then pulverized with Sichuan peppercorns using a low-temperature airflow pulverizer to a particle size D90≤150μm to obtain garlic micro powder and Sichuan peppercorn micro powder; Charcoal is ultra-finely pulverized under inert gas protection and passed through an 800-mesh sieve to obtain ultra-fine charcoal powder, which is then placed in a moisture-proof container for later use; S2. Stepwise dynamic countercurrent extraction: The herbal powder obtained in step S1 and all the sweet potato vinegar pulp were put into a multi-functional hot reflux extraction tank. Stirring was turned on, and the temperature was raised to 50±2℃ at a rate of 1.5℃ / min. The temperature was maintained and ultrasonically assisted extraction was performed at a power density of 50W / L for 45min. The temperature was then raised to 68±2℃ at a rate of 1℃ / min. Dynamic countercurrent circulation extraction was performed at this temperature for 4 hours. During the extraction process, a slight negative pressure of -0.02 to -0.03MPa was maintained in the tank. After the extraction was completed, the liquid was cooled to below 40℃ through the built-in cooling coil to obtain the primary alcohol extract. S3. Integrated extraction of flavor and heat-sensitive components: The fresh material to be used, garlic powder and Sichuan pepper powder obtained in step S1 are added together to the primary alcohol extract and warm-soaked for 90 min at 60±2℃ and under sealed conditions with a stirring speed of 120 rpm. S4. Precision separation and purification: The mixture after step S3 is fed into a tubular centrifuge and solid-liquid separation is performed under a centrifugal force of 10000G. The supernatant is collected. The supernatant is then filtered sequentially through a 50μm bag filter and a 5μm precision filter to obtain a clear extract. S5. Controllable homogenization and stabilization: The clarified extract is introduced into a jacketed cooling mixing tank, cooled and maintained at 25±3℃; under continuous stirring at 80rpm / min, sesame oil and honey are slowly and evenly added to the tank, and stirred for 20min until completely emulsified and dispersed; with the high-speed shear emulsifier running at 10000rpm / min, the ultrafine charcoal powder obtained in step S1 is evenly fed into the emulsifier head through a vacuum feeding device, and homogenized continuously for 15min to form a stable suspension system; S6. Aseptic filling: The homogenized semi-finished product is filtered through a 0.45μm terminal sterilization filter and then transferred to an aseptic temporary storage tank; using a light-proof and well-sealed pharmaceutical nasal drop bottle or spray bottle, the product is filled and capped in a Class B clean environment to obtain the finished herbal composition for nasal care.

4. The method for preparing the herbal composition for nasal care according to claim 3, characterized in that, The dynamic countercurrent circulation extraction in step S2 is achieved by pumping the liquid from the bottom of the extraction tank to the spray device on the top of the tank to form a continuous internal circulation, with a circulation flow rate of 2-3 times the tank volume per hour.

5. The method for preparing the herbal composition for nasal care according to claim 3, characterized in that, In step S4, after the tubular centrifugation, the obtained solid residue is rinsed with 30% of its mass of purified water. The rinsing liquid is collected separately, concentrated at low temperature, and then added back to the supernatant to improve the transfer rate of the target component.

6. The method for preparing the herbal composition for nasal care according to claim 3, characterized in that, In step S5, the sesame oil is subjected to nitrogen bubbling deoxygenation treatment for 10 minutes before use, and the honey is filtered through a 40-mesh sieve to remove crystallized particles and impurities.

7. The method for preparing the herbal composition for nasal care according to claim 3, characterized in that, In step S5, before being inhaled, the ultrafine charcoal powder is premixed with an equal weight of clarified extract to form a slurry, which is then quantitatively delivered to the emulsifying head via a peristaltic pump.

8. The method for preparing the herbal composition for nasal care according to claim 3, characterized in that, In step S6, before filling, high-purity nitrogen is introduced into the sterile temporary storage tank to replace the air at the top of the tank for 10 minutes, and the tank is kept at a slightly positive nitrogen pressure of 0.03-0.05 MPa until filling is completed.

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