An oral preparation with throat-clearing and throat-moistening efficacy and a preparation method thereof

Abstract

The application discloses an oral preparation with throat clearing and soothing efficacy and a preparation method thereof, and relates to the oral preparation field. The oral preparation comprises the following raw materials: honeysuckle, chrysanthemum, momordica grosvenori, figwort, dark plum, peppermint, black pepper extract powder, sweetening agent, beta-cyclodextrin, xylitol, potassium sorbate, citric acid, vitamin C sodium and purified water; the sweetening agent is prepared by compounding ammonium glycyrrhizinate and mogroside in a mass ratio of 8-20:1. The preparation method comprises the following steps: mixing the raw materials and performing enzymatic hydrolysis treatment; preparing a concentrated solution; preparing an auxiliary material solution; mixing the concentrated solution and the auxiliary material solution, and then performing two-stage filtration in sequence to obtain the target oral preparation. The content of chlorogenic acid, total flavones and total saponins in the product is increased by more than 50% compared with that in the traditional process, the oral bioavailability of chlorogenic acid and total flavones is effectively improved, the taste score reaches 37.8 points, the turbidity of the finished product is less than 0.5 NTU, the retention rate of chlorogenic acid after sterilization reaches 96.0%, and the shelf life reaches 24 months.

Description

Technical Field

[0001] This invention relates to the field of oral preparations technology, specifically to an oral preparation with throat-soothing and moisturizing effects and its preparation method. Background Technology

[0002] Throat discomfort, such as dryness, pain, a feeling of something stuck in the throat, and hoarseness, is a common health problem among modern people, especially teachers, broadcasters, singers, and others who frequently use their voices, as well as those who spend long periods in air-conditioned or polluted environments. Currently, throat-soothing products on the market mainly exist in the form of lozenges, syrups, and oral liquids, but their poor taste and inconvenience in carrying them are significant drawbacks.

[0003] Traditional extraction processes result in significant losses of heat-sensitive active ingredients. Most existing throat-soothing products employ traditional water-decoction extraction, which involves long extraction times (typically 1-2 hours each time) and high temperatures (100℃ at normal pressure boiling). This leads to substantial degradation or volatilization of heat-sensitive components such as chlorogenic acid, flavonoids, and peppermint volatile oils during extraction. This not only reduces raw material utilization and weakens the product's throat-soothing efficacy but also forces manufacturers to compensate for the losses by increasing the amount of raw materials used, thus raising production costs.

[0004] The taste is poor and there is an over-reliance on sucrose or artificial sweeteners. Traditional Chinese medicine extracts generally suffer from a strong bitter taste, a pronounced medicinal flavor, and an unpleasant mouthfeel. To mask this unpleasant taste, existing products often add large amounts of sucrose sweeteners such as white sugar and glucose syrup, or use artificial sweeteners such as sucralose and aspartame. Sucrose sweeteners are high in calories and are not suitable for long-term consumption by diabetics or those trying to control their blood sugar. While artificial sweeteners are calorie-free, their safety has been controversial in recent years, leading to significant consumer aversion. Current technology does not integrate the selection of sweeteners with enhancing throat-soothing effects; sweeteners are used merely as flavoring agents and fail to exert a synergistic pharmacological effect.

[0005] The product has poor clarity and insufficient shelf-life stability. Existing oral formulations have low filtration precision, mostly using 200-mesh sieves or filter bags with a filter size of a few micrometers. This makes it difficult to effectively remove turbid substances such as colloids, pectin, and fine suspended particles from the extract. As a result, the product is prone to problems such as precipitation, flocculation, or darkening of color during its shelf life, which seriously affects the product's appearance quality and consumer acceptance.

[0006] In conclusion, developing an oral preparation for relieving sore throat with high extraction rate of effective ingredients, excellent taste, no artificial sweeteners, and high clarity has significant market value and practical significance. Summary of the Invention

[0007] The purpose of this invention is to provide an oral preparation with throat-soothing and moisturizing effects and its preparation method. Through the synergistic effect of enzymatic pretreatment and ultrasound-assisted extraction, the extraction rate of active ingredients such as chlorogenic acid, total flavonoids, and total saponins is significantly improved, resulting in a substantial increase in the content of effective components in the product. The use of peppermint oil β-cyclodextrin inclusion technology and a compound natural sweetener of ammonium glycyrrhizate and mogroside effectively masks the bitterness of the traditional Chinese medicine, retains a refreshing aroma, and imparts a lasting sweet aftertaste, significantly improving the product's taste. The addition of black pepper extract enhances the bioavailability of the active ingredients in vivo, further improving the throat-soothing effect. Through two-stage microfiltration, an acidic antioxidant environment constructed with sodium vitamin C and citric acid, and nitrogen protection before filling, combined with an optimized short-time sterilization process, the product's clarity, stability, and shelf life are significantly improved. This systematically solves the technical problems of low effective component content, unpleasant taste, and poor stability in existing throat-soothing and moisturizing oral preparations.

[0008] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0009] A method for preparing an oral preparation with throat-soothing and moisturizing effects includes the following steps:

[0010] S100. Add cellulase and pectinase to the pretreated mixed raw materials of honeysuckle, chrysanthemum, monk fruit, scrophularia and dried plum, moisten with purified water, and carry out enzymatic hydrolysis.

[0011] S200. Add purified water to the enzymatically hydrolyzed material, perform two ultrasonic extractions, combine the two extraction filtrates, filter, and obtain the extract; add sodium vitamin C to the extract, stir evenly, concentrate under reduced pressure until the relative density is 1.02~1.10, and obtain the concentrate.

[0012] S300. Peppermint oil is extracted by steam distillation. β-Cyclodextrin is dissolved in purified water, and the peppermint oil is added dropwise to prepare a peppermint oil-β-cyclodextrin inclusion complex solution. Black pepper extract powder, sweetener, xylitol, potassium sorbate and citric acid are added sequentially to the above peppermint oil-β-cyclodextrin inclusion complex solution and stirred until completely dissolved to obtain an excipient solution.

[0013] S400. Mix the concentrated solution and excipient solution evenly, add the remaining purified water, stir evenly to obtain the preparation solution, and filter the preparation solution in two stages to obtain the target oral formulation.

[0014] In the preparation method of this invention, a tight synergistic chain is formed between the various steps.

[0015] The enzymatic hydrolysis process in S100 selectively disrupts the cell walls of raw materials such as honeysuckle, chrysanthemum, monk fruit, scrophularia, and dried plum, resulting in a looser cell structure. The ultrasound-assisted extraction in S200 utilizes cavitation to further enhance mass transfer. The synergistic effect of these two processes increases the extraction rate of active ingredients by more than 50% compared to traditional water decoction, while maintaining the extraction temperature at 75-82℃, significantly reducing the degradation of heat-sensitive components. Simultaneously, the sodium vitamin C added before vacuum concentration in S200, combined with the acidic environment (pH 4.5±0.3) created by citric acid in the subsequent S400 process, forms an antioxidant relay, and together with two-stage microfiltration to remove pro-oxidative particles, increases chlorogenic acid retention by 30%, extending the product's shelf life to 18 months.

[0016] Traditional processes involve directly adding peppermint to the extraction tank, resulting in significant loss of volatile oils and the dissolution of water-soluble bitter substances. This invention separates peppermint from the main extraction system. The S300 extractor uses the same enzymatic hydrolysis parameters as the S100 system and ultrasonic conditions compatible with the S200 system to extract the volatile oils separately. The extracted oils are then protected with β-cyclodextrin inclusion complexes. The resulting inclusion complex solution is directly used for mixing and volume adjustment in the S400 system. This integrated design of separation, extraction, inclusion complexation, and backfilling preserves the refreshing aroma, avoids bitterness, simplifies the process, and solves the long-standing taste problem of throat-soothing oral preparations.

[0017] The S400 employs a two-stage filtration system: pre-filtration (filter bag) and terminal filtration (microporous membrane). The filtration precision is improved from the traditional 200-mesh sieve to 0.22~0.65μm, effectively removing colloids, fine particles, and most microorganisms, giving the product a highly clear appearance, while creating favorable conditions for subsequent nitrogen filling and short-time sterilization.

[0018] The interconnected synergistic effects described above enable this invention to significantly outperform existing technologies in four dimensions: extraction efficiency, retention of active ingredients, taste quality, and shelf stability.

[0019] Furthermore, an oral preparation with throat-soothing and moisturizing effects, by weight,

[0020] Honeysuckle 10-15 parts, chrysanthemum 8-12 parts, monk fruit 7-11 parts, scrophularia 6-10 parts, dried plum 6-10 parts, peppermint 3-6 parts, black pepper extract powder 0.005-0.01 parts, sweetener 0.4-1.0 parts, β-cyclodextrin 0.8-1.5 parts, xylitol 5-15 parts, potassium sorbate 0.3-0.8 parts, citric acid 0.2-0.5 parts, sodium vitamin C 0.02-0.08 parts, purified water 1765-1915 parts;

[0021] The sweetener is composed of ammonium glycyrrhizate and mogroside in a mass ratio of 8 to 20:1.

[0022] This invention breaks through the old model of traditional throat-clearing oral preparations that rely on sucrose or artificial sweeteners and use simple decoction. For the first time, it integrates natural sweeteners (ammonium glycyrrhizate, mogroside), black pepper extract and β-cyclodextrin system to form a new solution that combines enhanced efficacy, optimized taste and stable ingredients.

[0023] The natural sweetener is a blend of ammonium glycyrrhizate and mogroside in a ratio of 8-20:1, completely replacing sucrose and artificial sweeteners. Both have anti-inflammatory, antitussive, and throat-soothing pharmacological activities. Together with the dried plum in the formula, they form a classic combination of sour and sweet flavors in traditional Chinese medicine, greatly enhancing the effects of promoting saliva production and moisturizing the throat.

[0024] Although black pepper extract (piperine) is not a direct throat-clearing component, as a natural bioavailability enhancer, when added in extremely low doses, it significantly improves the oral absorption rate of chlorogenic acid in honeysuckle and flavonoids in chrysanthemum by inhibiting the activity of glucuronyl transferase (UGT) in the intestine and liver, thus greatly enhancing the throat-clearing efficacy with the same amount of ingredients.

[0025] Peppermint volatile oils (menthol, etc.) are the core ingredients for a cooling and soothing effect on the throat, but they are volatile, poorly water-soluble, and irritating. β-cyclodextrin stabilizes these components through inclusion complexation technology, solving the industry-wide problems of aroma loss, insufficient cooling sensation, and irritating taste during product processing and storage. Sodium vitamin C and citric acid work together to create an acidic environment, protecting heat-sensitive phenolic acids such as chlorogenic acid and extending shelf life.

[0026] The above-mentioned ingredients exert their throat-soothing and moisturizing effects through multiple synergistic mechanisms: Ume plum and ammonium glycyrrhizate nourish yin, relieving dry throat and thirst; black pepper extract enhances the bioavailability of chlorogenic acid and flavonoids, increasing the anti-inflammatory effect in vivo by 2-5 times; monk fruit and ammonium glycyrrhizate complement each other's antitussive and expectorant targets, synergistically relieving itchy throat and dry cough; Scrophularia ningpoensis and ume plum nourish yin, reduce heat, promote saliva production, and astringe, targeting throat dryness caused by deficiency heat. Simultaneously, β-cyclodextrin inclusion complexation increases the water solubility of peppermint oil, reduces volatility, and moderates irritation, reducing aroma loss by more than 50%; sodium vitamin C and citric acid inhibit oxidation reactions, protecting heat-sensitive components. These synergistic effects make this invention significantly superior to existing technologies in terms of throat-soothing efficacy, palatability, and product stability.

[0027] Further, in steps S100 to S400, the ratio of purified water used is 1:18.5~25.5:3.5~4.5:17~23; wherein, in step S200, in the two extraction processes, the ratio of purified water used in the first extraction and the second extraction is 1.3~1.6:1.

[0028] If the amount of wetting water in the enzymatic hydrolysis is too low, enzyme activity will be limited; if it is too high, the substrate will be diluted. Matching the extraction water ratio with the ultrasonic-insulation parameters allows the dissolution rate of the active ingredient to reach its peak while minimizing the energy consumption for concentration. The difference in water volume between the two extractions (approximately 1.3 to 1.6 times) precisely corresponds to the kinetic law that the first extraction dissolves 60-70% of the components, while the second extraction dissolves the remaining components. This ratio system ensures batch-to-batch reproducibility.

[0029] Further, in step S100, the mass ratio of cellulase, pectinase and mixed raw materials is 0.02~0.08:0.01~0.05:1; the enzymatic hydrolysis temperature is 35~42℃, and the enzymatic hydrolysis time is 25~35 minutes.

[0030] This range represents a balance between maintaining high enzyme activity and avoiding pre-degradation of heat-sensitive components. It also requires extremely low enzyme dosage, effectively disrupting cell walls without introducing off-flavors or increasing costs. Furthermore, it can increase the extraction rates of chlorogenic acid from honeysuckle and total flavonoids from chrysanthemum by more than 20% compared to conventional enzymatic hydrolysis.

[0031] Further, in step S200, purified water is added to the enzymatically hydrolyzed material, and the first extraction is performed under ultrasound at 35~45kHz and 240~350W for 25~35 minutes, followed by heating to 75~82℃ and holding for 0.8~1.2 hours.

[0032] Add purified water to the filter residue and perform a second extraction under the same conditions, using ultrasound for 18-25 minutes, followed by incubation at 75-80℃ for 0.5-1 hour. Filter both extractions through a 180-220 mesh filter. Combine the filtrates from both extractions to obtain the extract.

[0033] The first extraction takes 25-35 minutes and is kept warm for 0.8-1.2 hours, while the second extraction takes 18-25 minutes and is kept warm for 0.5-1 hour. This gradient design keeps the total extraction time within 2 hours, which is 50% shorter than traditional decoction.

[0034] Further, in step S200, sodium vitamin C is added to the extract and concentrated under reduced pressure at 65~72℃ and -0.08~0.02MPa to a relative density of 1.02~1.1 to obtain a concentrated solution.

[0035] This temperature is lower than that of conventional vacuum concentration. Combined with a higher vacuum level and the antioxidant protection of sodium vitamin C, the degradation rate of chlorogenic acid during the concentration process is reduced by about 30%. The concentrate with a relative density of 1.02~1.1 has good fluidity, which facilitates subsequent mixing with excipient solutions and prevents precipitation due to over-concentration.

[0036] Further, in step S300, the method for extracting peppermint oil by steam distillation includes the following: taking the prescribed amount of peppermint raw material, pulverizing it at low temperature, adding cellulase and pectinase, wetting it with water, and enzymatically hydrolyzing it at 35~45℃ for 20~40 minutes; subjecting the enzymatically hydrolyzed material to ultrasonic-assisted treatment for 10~15 minutes; then performing steam distillation for 1.5~3.0 hours, collecting the condensed distillate, allowing it to stand and separate into layers, and collecting the upper peppermint oil layer;

[0037] The mass ratio of cellulase, pectinase and peppermint raw material is 0.03-0.08:0.01-0.05:1, and the mass ratio of water (added separately and not included in the original 1765-1915 parts) to peppermint raw material is 8-11:1.

[0038] The water used in the steam distillation method for extracting peppermint oil is purified water. This water is added separately and is not included in the 1765-1915 parts of purified water in the formula. The three-stage enzymatic hydrolysis-ultrasound-distillation scheme of this invention utilizes existing equipment resources in the main process to fully disrupt the peppermint cell walls, making it easier for the oil glands to release volatile oils.

[0039] Further, in step S300, peppermint oil is added to the β-cyclodextrin aqueous solution at a rate of 20-40 drops / min, and stirred at 38-42°C for 25-35 min to obtain a peppermint oil-β-cyclodextrin inclusion complex solution.

[0040] The conditions are mild (close to room temperature) and short (≤35 minutes), far superior to traditional inclusion processes (which often require several hours). The moderate dripping rate ensures sufficient contact and uniform dispersion of the oil and cyclodextrin. After inclusion, the water solubility of peppermint oil is improved, its volatility is reduced, and its aroma retention rate is increased by more than 50%. Furthermore, it can be directly used to prepare excipient solutions without additional separation, achieving integrated inclusion-dissolution-mixing and solving the problems of uneven peppermint oil addition and easy loss in industrial production.

[0041] Further, in step S400, the two-stage filtration includes the following: the prepared solution is filtered sequentially through a 2-10 μm filter bag and a 0.22-0.65 μm microporous membrane to obtain the target oral formulation.

[0042] 2-10μm filter bags remove large suspended solids and some colloids, protecting the subsequent microporous membrane from clogging; 0.22-0.65μm microporous membranes intercept fine particles, microorganisms, and large colloids, keeping the product turbidity below 0.5 NTU and preventing sedimentation during the shelf life. Compared to traditional 200-mesh sieves, the filtration precision is improved by two orders of magnitude, and no chemical clarifying agents are needed, significantly improving the sensory quality and hygiene safety of the product.

[0043] Furthermore, the prepared target oral formulation is purged with nitrogen before filling and sealing; after sealing, it is sterilized by autoclaving at 110~120℃ for 28~33 minutes.

[0044] Nitrogen purging reduces the residual oxygen in the headspace of the tank to below 1%, synergistically forming a gas-liquid dual-phase protection with the liquid-phase antioxidant effect of sodium vitamin C in the formula, effectively inhibiting the oxidative browning of chlorogenic acid and total flavonoids. The sterilization temperature of 110~120℃ and the time of 28~33 minutes are 25% shorter than the conventional 40 minutes, thanks to the significant reduction in the initial bacterial count by the preceding enzymatic hydrolysis, ultrasonication, and microfiltration steps, and the acidic pH also inhibiting heat-resistant bacteria.

[0045] An oral preparation with throat-soothing and moisturizing effects, prepared by the method described above.

[0046] Compared with the prior art, the beneficial effects of the present invention are:

[0047] 1. This invention utilizes enzymatic hydrolysis-ultrasonic low-temperature extraction, increasing the content of chlorogenic acid, total flavonoids, and total saponins by over 50% compared to traditional processes, reaching 29.6, 40.1, and 16.9 mg / 100mL respectively. Simultaneously, the use of ammonium glycyrrhizate and mogroside (8-20:1) completely replaces artificial sweeteners, achieving a taste score of 37.8 points. Combined with dried plum, it creates a sweet and sour flavor that nourishes the body and moistens the throat. The peppermint oil, after enzymatic hydrolysis-ultrasonic extraction-distillation-β-cyclodextrin encapsulation, achieves a 91% retention rate, providing a long-lasting and mild cooling sensation. Two-stage microfiltration combined with nitrogen filling and sodium VC for antioxidant effects results in a finished product turbidity <0.5 NTU. After sterilization, the chlorogenic acid retention rate is 96.0%, and the shelf life reaches 24 months.

[0048] 2. Clinical trials showed that the improvement rates of the present invention for the main symptoms of dry throat, sore throat, itchy throat, dry cough, foreign body sensation in the throat, and aggravation of excessive talking were 86.1%, 71.7%, 76.5%, 70.3%, 81.5%, and 95.5%, respectively. The differences in the total symptom score and the total sign score before and after the trial were extremely significant (P<0.001). The overall effective rate was 46.6%, which was significantly better than the control group (1.9%, P<0.001), and no adverse reactions were observed.

[0049] 3. This invention increases the oral bioavailability of chlorogenic acid by 131% and the oral bioavailability of total flavonoids by 117% by 60% to 80% without increasing the amount of active ingredients by adding extremely low doses of black pepper extract powder to the formula. Attached Figure Description

[0050] Figure 1 This is a diagram of a rat paw swelling experiment.

[0051] Figure 2 This is a diagram of a mouse ear swelling experiment.

[0052] Figure 3 Image of a person's throat before taking this product;

[0053] Figure 4 This is a diagram of the throat of a person after taking this product. Detailed Implementation

[0054] The present invention will now be further described.

[0055] Example 1

[0056] Weigh the following ingredients: 125g honeysuckle, 100g chrysanthemum, 90g monk fruit, 80g scrophularia, 80g dried plum, 45g peppermint, 0.075g black pepper extract powder (CAS No. 94-62-2), 6.5g ammonium glycyrrhizate, 0.5g monk fruit glycoside, 11.5g β-cyclodextrin, 100g xylitol, 5.5g potassium sorbate, 3.5g citric acid, 0.5g sodium vitamin C, and 18800g purified water (i.e., 18.8L).

[0057] A method for preparing an oral preparation with throat-soothing and moisturizing effects includes the following steps:

[0058] S100: Grind honeysuckle and chrysanthemum at low temperature and pass through a 20-mesh sieve; crack the shell of monk fruit, cut scrophularia into pieces, and remove the pits from dried plums and cut them into pieces; set aside the mint. Mix the mixed ingredients except the mint with 0.5g of cellulase and 0.3g of pectinase, add 400g of purified water to moisten, and enzymatically hydrolyze at 40℃ for 30 minutes.

[0059] S200: Add 5200g of purified water to the enzymatic hydrolysate, extract under ultrasonication at 40kHz and 295W for 30 minutes, heat to 80℃ and hold for 1 hour, then filter. Add 3600g of purified water to the residue, extract under the same conditions under ultrasonication for 21.5 minutes, hold at 80℃ for 0.75 hours, then filter. Both extractions are filtered through a 200-mesh filter; combine the two filtrates to obtain the extract. Add 0.5g of sodium vitamin C to the extract, concentrate under reduced pressure at 68.5℃ and -0.05MPa to a relative density of 1.06 (measured at 60℃) to obtain the concentrate.

[0060] S300: Take 45g of peppermint, pulverize at low temperature, add 0.025g of cellulase and 0.0135g of pectinase, moisten with water, and enzymatically hydrolyze at 40℃ for 30 minutes; then sonicate for 12.5 minutes (40kHz, 295W); then add 427.5g of purified water (water to peppermint mass ratio 9.5:1), steam distill for 2.5 hours, condense and collect the distillate, let it stand to separate the layers, and collect the upper layer of peppermint oil, 1.1g. Separately, take 1600g of purified water, add 11.5g of β-cyclodextrin to dissolve, add peppermint oil dropwise at a rate of 30 drops / min, stir at 40℃ for 30 minutes to prepare an inclusion complex solution. Add 0.075g of black pepper extract powder, 6.5g of ammonium glycyrrhizate, 0.5g of mogroside, 100g of xylitol, 5.5g of potassium sorbate, and 3.5g of citric acid to the solution in sequence, stir to dissolve, and obtain the excipient solution.

[0061] S400: Mix the concentrate and excipient solution, add 8000g of purified water, and stir until homogeneous. Filter sequentially through a 6μm filter bag and a 0.435μm microporous membrane to obtain the target oral formulation. Purge with nitrogen before filling and sealing, and sterilize by autoclaving at 115℃ for 30.5 minutes after sealing.

[0062] Example 2

[0063] Weigh the following ingredients: 100g honeysuckle, 80g chrysanthemum, 70g monk fruit, 60g scrophularia, 60g dried plum, 30g peppermint, 0.05g black pepper extract powder (CAS No. 94-62-2), 3.6g ammonium glycyrrhizate, 0.4g monk fruit glycoside, 8g β-cyclodextrin, 50g xylitol, 3g potassium sorbate, 2g citric acid, 0.2g sodium vitamin C, and 17650g purified water (i.e., 17.65L).

[0064] A method for preparing an oral preparation with throat-soothing and moisturizing effects includes the following steps:

[0065] S100: Grind honeysuckle and chrysanthemum at low temperature and pass through a 20-mesh sieve; crack the shell of monk fruit, cut scrophularia into pieces, and remove the pits from dried plums and cut them into pieces; set aside peppermint separately. Mix the mixed raw materials except peppermint with 0.2g of cellulase and 0.1g of pectinase, add 440g of purified water to moisten, and enzymatically hydrolyze at 35℃ for 25 minutes.

[0066] S200: Add 4620g of purified water to the enzymatic hydrolysate, extract under ultrasonication at 35kHz and 240W for 25 minutes, heat to 75℃ and hold for 0.8 hours, then filter. Add 3550g of purified water to the filter residue, extract under the same conditions under ultrasonication for 18 minutes, and hold at 75℃ for 0.5 hours. Filter. Both extractions are filtered through a 180-mesh filter; combine the two filtrates to obtain the extract. Add 0.2g of sodium vitamin C to the extract, concentrate under reduced pressure at 65℃ and -0.08MPa to a relative density of 1.02 (measured at 60℃) to obtain the concentrate.

[0067] S300: Take 30g of peppermint, pulverize at low temperature, add 0.009g of cellulase and 0.003g of pectinase, moisten with water, and enzymatically hydrolyze at 35℃ for 20 minutes; then sonicate for 10 minutes (35kHz, 240W); then add 240g of purified water (water to peppermint mass ratio 8:1), steam distill for 1.5 hours, condense and collect the distillate, let it stand to separate the layers, collect the upper layer of peppermint oil, 0.70. Separately, take 1540g of purified water, add 8g of β-cyclodextrin to dissolve, add peppermint oil dropwise at a rate of 20 drops / min, stir at 38℃ for 25 minutes to prepare an inclusion complex solution. Add 0.05g of black pepper extract powder, 3.6g of ammonium glycyrrhizate, 0.4g of mogroside, 50g of xylitol, 3g of potassium sorbate, and 2g of citric acid to it in sequence, stir to dissolve, and obtain the excipient solution.

[0068] S400: Mix the concentrate and excipient solution, add 7500g of purified water, and stir until homogeneous. Filter sequentially through a 2μm filter bag and a 0.22μm microporous membrane to obtain the target oral formulation. Purge with nitrogen before filling and sealing, and sterilize by autoclaving at 110℃ for 28 minutes after sealing.

[0069] Example 3

[0070] Weigh the following ingredients: 150g honeysuckle, 120g chrysanthemum, 110g monk fruit, 100g scrophularia, 100g dried plum, 60g peppermint, 0.1g black pepper extract powder (CAS No. 94-62-2), 9.5g ammonium glycyrrhizate, 0.5g monk fruit glycoside, 15g β-cyclodextrin, 150g xylitol, 8g potassium sorbate, 5g citric acid, 0.8g sodium vitamin C, and 19150g purified water (i.e., 19.15L).

[0071] A method for preparing an oral preparation with throat-soothing and moisturizing effects includes the following steps:

[0072] S100: Low-temperature pulverize honeysuckle and chrysanthemum and pass through a 20-mesh sieve; crack the shell of monk fruit, cut scrophularia into pieces, and remove the pits from dried plums and cut them into pieces; set aside peppermint separately. Mix the mixed raw materials except peppermint with 0.8g of cellulase and 0.5g of pectinase, add 354.6g of purified water to moisten, and enzymatically hydrolyze at 42℃ for 35 minutes.

[0073] S200: Add 5564.5g of purified water to the enzymatic hydrolysate, extract under ultrasonication at 45kHz and 350W for 35 minutes, heat to 82℃ and hold for 1.2 hours, then filter. Add 3477.8g of purified water to the filter residue, extract under the same conditions under ultrasonication for 25 minutes, hold at 80℃ for 1 hour, then filter. Both extractions were filtered through a 220-mesh filter; combine the two filtrates to obtain the extract. Add 0.8g of sodium vitamin C to the extract, concentrate under reduced pressure at 72℃ and -0.02MPa to a relative density of 1.10 (measured at 60℃) to obtain the concentrate.

[0074] S300: Take 60g of peppermint, pulverize at low temperature, add 0.048g of cellulase and 0.03g of pectinase, moisten with water, and enzymatically hydrolyze at 45℃ for 40 minutes; then sonicate for 15 minutes (45kHz, 350W); then add 660g of purified water (water to peppermint mass ratio 11:1), steam distill for 3.0 hours, condense and collect the distillate, let it stand to separate the layers, and collect 1.5g of the upper layer of peppermint oil. Separately, take 1595.7g of purified water, add 15g of β-cyclodextrin to dissolve, add peppermint oil dropwise at a rate of 40 drops / min, stir at 42℃ for 35 minutes to prepare an inclusion complex solution. Add 0.1g of black pepper extract powder, 9.5g of ammonium glycyrrhizate, 0.5g of mogroside, 150g of xylitol, 8g of potassium sorbate, and 5g of citric acid to it in sequence, stir to dissolve, and obtain the excipient solution.

[0075] S400: Mix the concentrate and excipient solution, add 8155.8g of purified water, and stir until homogeneous. Filter sequentially through a 10μm filter bag and a 0.65μm microporous membrane to obtain the target oral formulation. Purge with nitrogen before filling and sealing, and sterilize by autoclaving at 120℃ for 33 minutes after sealing.

[0076] Comparative Example 1

[0077] The traditional throat-clearing oral preparation process is adopted: the raw materials are not subjected to enzymatic hydrolysis and ultrasonic treatment. They are directly added to 10 times the volume of purified water, decocted at 100°C for 2 hours, and filtered. The residue is then added to 8 times the volume of water, decocted at 100°C for 1.5 hours, and the filtrates are combined and concentrated under reduced pressure to the same relative density. Peppermint is directly decocted with other raw materials (volatile oil is not extracted separately). β-cyclodextrin inclusion is not used; black pepper extract powder is added directly after concentration. Filtration is performed using only a 200-mesh sieve, without two-stage microfiltration. After filling, it is sterilized at 116°C for 40 minutes without nitrogen purging. Other procedures are the same as in Example 1.

[0078] Comparative Example 2

[0079] No β-cyclodextrin was added to the raw materials. In step S300, peppermint oil was added directly to the excipient solution without being encapsulated by β-cyclodextrin. The rest was the same as in Example 1.

[0080] Comparative Example 3

[0081] The formula does not include black pepper extract powder, but otherwise it is the same as in Example 1.

[0082] Comparative Example 4

[0083] No sodium vitamin C was added; otherwise, it was the same as in Example 1.

[0084] Comparative Example 5

[0085] Citric acid was not added; otherwise, it was the same as in Example 1.

[0086] Comparative Example 6

[0087] The sweetener used in the formula is only ammonium glycyrrhizate (the total mass of sweetener remains unchanged), and the rest is the same as in Example 1.

[0088] Comparative Example 7

[0089] The only sweetener used in the formula is mogroside (total mass of sweetener remains unchanged), and the rest is the same as in Example 1.

[0090] Comparative Example 8

[0091] In step S100, enzymatic hydrolysis is not performed. After the raw material is moistened, step S200 is performed directly, and the rest is the same as in Example 1.

[0092] Comparative Example 9

[0093] In step S200, ultrasonic treatment is not performed; only heating and heat preservation are performed for extraction. The rest is the same as in Example 1.

[0094] Comparative Example 10

[0095] Meanwhile, the enzymatic pretreatment of S100 and the ultrasonic-assisted extraction of S200 are omitted (only 80℃ water bath extraction is used), and the rest is the same as in Example 1.

[0096] Comparative Example 11

[0097] In step S200, purified water (the sum of the water volumes from the two extractions) is added to the enzymatically hydrolyzed material, and extraction is performed under ultrasonication at 40 kHz and 295 W for 51.5 minutes. The temperature is then raised to 80°C and maintained for 1.75 hours. The extract is then filtered through a 200-mesh sieve to obtain the extract. The rest is the same as in Example 1.

[0098] Comparative Example 12

[0099] Nitrogen purging is not performed before filling and sealing; otherwise, it is the same as in Example 1.

[0100] Comparative Example 13

[0101] Weigh the following ingredients: 10g honeysuckle, 8g chrysanthemum, 7g monk fruit, 6g scrophularia, 6g dried plum, 3g peppermint, 15g xylitol, 0.5g potassium sorbate, 0.15g sucralose, and appropriate amount of purified water, add to 1000mL.

[0102] A method for preparing an oral preparation with throat-soothing and moisturizing effects includes the following steps:

[0103] 1. Raw material pretreatment: The monk fruit is crushed into large pieces and mixed with honeysuckle, chrysanthemum, scrophularia, dried plum and mint as extraction raw materials.

[0104] 2. Extraction and filtration: Boil twice with water, adding 10 times the amount of water each time and boiling for 2 hours. Filter through a 200-mesh sieve to obtain the extract.

[0105] 3. Vacuum Concentration: The extract is concentrated under reduced pressure, with controlled conditions of temperature 60~80℃ and vacuum degree -0.04~-0.08MPa, until the relative density is 1.02~1.10 (60±5℃), yielding the concentrated solution. The area within the dashed box represents a Class 100,000 cleanroom, and subsequent steps will be performed within this area.

[0106] 4. Preparation of excipients: Add xylitol, potassium sorbate and sucralose to 50wt% purified water and stir for 15 minutes to dissolve them to obtain a solution.

[0107] 5. Solution preparation and filtration: Mix the solution and concentrate, add water to the prepared volume, stir for 20 minutes to mix evenly, and obtain the prepared solution; then filter with a 10μm filter bag to obtain the solution to be filled.

[0108] 6. Sterilization: The canned oral preparations are sterilized by hot autoclaving at 116℃ for 40 minutes to obtain the product to be packaged.

[0109] The performance of the oral formulations prepared by the methods in Examples 1-3 and Comparative Examples 1-13 is shown in Table 1.

[0110] Test method:

[0111] Chlorogenic acid is an organic acid with characteristic ultraviolet absorption at a wavelength of 327 nm. HPLC can be used to effectively separate it from other phenolic acid components and then accurately quantify it.

[0112] Total flavonoids are composed of various flavonoid compounds. Under the colorimetric system of sodium nitrite-aluminum nitrate-sodium hydroxide, the flavonoid components complex with aluminum ions to form a red complex, which has the maximum absorption at 510 nm. The total flavonoid content is determined by colorimetry.

[0113] Total saponins are composed of various saponin compounds. In the vanillin-perchloric acid system, saponins and vanillin undergo dehydration condensation in perchloric acid to form a purple-red compound with maximum absorption at 544 nm. The total saponin content is determined by colorimetry.

[0114] Peppermint oil retention rate: The menthol content in the finished product was determined by steam distillation-gas chromatography, and the ratio to the theoretical amount added was calculated.

[0115] Turbidity: Measured using a turbidimeter (NTU).

[0116] Shelf life sedimentation: Accelerated test at 37℃ for 6 months, observe sedimentation.

[0117] Taste rating: 20 sensory evaluators conducted a blind taste test and rated the sweetness on four aspects (1-10 points each) based on the naturalness of the sweetness, the masking of bitterness, the persistence of the coolness, and the overall acceptability. The average value was then taken.

[0118] Table 1. Performance of oral formulations prepared by the methods of Examples 1-3 and Comparative Examples 1-13

[0119]

[0120] Note: The taste score is the sum of four factors: naturalness of sweetness, masking of bitterness, persistence of coolness, and overall acceptability (1-10 points for each factor, 40 points in total).

[0121] As shown in Table 1, Examples 1-3 exhibited significantly higher chlorogenic acid content (25.5-29.6 mg / 100mL), total flavonoid content (34.5-40.1 mg / 100mL), and total saponin content (14.5-16.9 mg / 100mL) compared to traditional processes. Furthermore, they demonstrated excellent peppermint oil retention (79.2%-91.0%), extremely low turbidity (0.28-0.45 NTU), and no precipitation was observed in accelerated testing. Taste scores ranged from 33.5 to 37.8, chlorogenic acid retention after sterilization was 91.2%-96.0%, and shelf life reached 18-24 months.

[0122] Comparative Example 1 used a traditional throat-clearing oral preparation process. Due to the high-temperature and long-term extraction, heat-sensitive components such as chlorogenic acid, flavonoids, and saponins were significantly degraded, and almost all of the peppermint volatile oil was lost. As a result, the contents of chlorogenic acid, total flavonoids, and total saponins decreased to 14.5 mg / 100 mL, 19.5 mg / 100 mL, and 8.0 mg / 100 mL, respectively, which were only about 50% of those in Example 1. The peppermint oil retention rate was only 18.3%, and the turbidity of the finished product was as high as 8.6 NTU. The taste score was only 18.5, with obvious bitterness and no cooling sensation. After sterilization, the chlorogenic acid retention rate was only 75.2%, the shelf life at room temperature was only 9 months, and a large amount of sediment appeared in the product. Comparative Example 1 fully demonstrates the comprehensive defects of the traditional high-temperature decoction, direct feeding, single-stage filtration, and long-term sterilization process.

[0123] The only difference between Comparative Example 2 and Example 1 is that β-cyclodextrin was not added; that is, peppermint oil was directly added to the excipient solution without inclusion complexation. Free peppermint oil is easily volatile, oxidized, and aggregated in oral formulations, resulting in a sharp drop in peppermint oil retention to 52.6%, insufficient cooling sensation, and a spicy, irritating taste, with a taste score decreasing to 26.3. Simultaneously, oil droplet aggregation slightly increased turbidity to 0.35 NTU, shortening the shelf life to 18 months. Comparative Example 2 confirms that β-cyclodextrin inclusion complexation is a key technology for solving the loss of peppermint aroma and the harshness of the taste.

[0124] Comparative Example 3's formulation did not include black pepper extract powder; otherwise, it was identical to Example 1. Since black pepper extract does not affect the extraction, filtration, or sterilization processes, nor does it alter the physicochemical properties and taste of the oral preparation, its physicochemical indicators are essentially the same as in Example 1: chlorogenic acid 27.8 mg / 100 mL, peppermint oil retention 86.0%, turbidity 0.33 NTU, and taste score 35.8. The role of black pepper extract is to improve the bioavailability of components such as chlorogenic acid in vivo. Since efficacy data is not shown in the table, it is reasonable that its content and retention rate are similar to those of Example 1.

[0125] In Comparative Example 4, due to the lack of antioxidant protection, dissolved oxygen and free radicals accelerated the oxidative degradation of phenolic acids such as chlorogenic acid during vacuum concentration and high-temperature sterilization, resulting in a decrease in chlorogenic acid content to 22.5 mg / 100 mL. After sterilization, the chlorogenic acid retention rate was only 81.3%, and the shelf life was shortened to 16 months. This comparative example demonstrates that antioxidant protection of sodium vitamin C is an essential measure to ensure the retention rate of active ingredients and color stability.

[0126] In Comparative Example 5, chlorogenic acid stability decreased under a slightly alkaline environment, making it more susceptible to isomerization and oxidation. Furthermore, the solubility of macromolecules such as pectin and protein decreased, leading to their aggregation and precipitation. Consequently, the chlorogenic acid content decreased to 24.0 mg / 100 mL, the turbidity increased to 0.52 NTU, a small amount of precipitation appeared, and the chlorogenic acid retention rate after sterilization was 88.2%, with the taste score slightly decreasing to 35.5. This confirms that citric acid, by adjusting pH, not only improves flavor but is also crucial for ensuring product stability and antioxidant effects.

[0127] Comparative Example 6 replaced the sweetener with ammonium glycyrrhizate alone (total sweetness remained unchanged), with the rest being the same as in Example 1. The physicochemical properties (chlorogenic acid 27.8 mg / 100 mL, turbidity 0.33 NTU, etc.) were similar to those of Example 1, but due to the prominent licorice flavor and aftertaste of ammonium glycyrrhizate, the sweetness was not pure enough, and its ability to mask bitterness decreased, resulting in a taste score of 31.2. This indicates that ammonium glycyrrhizate alone cannot achieve the ideal taste of the compound formulation.

[0128] Comparative Example 7 replaced the sweetener with mogroside alone (total sweetness remained unchanged), otherwise it was the same as in Example 1. The physicochemical properties were similar to those of Example 1, but mogroside exhibited rapid sweetness decay, lacked a lingering aftertaste, and had a weak ability to mask bitterness, resulting in a taste score of only 30.5, lower than Comparative Example 6. Comparative Examples 6 and 7 together verified the synergistic and complementary effect of the combination of two natural sweeteners.

[0129] Comparative Example 8 omitted enzymatic pretreatment and only underwent ultrasound-assisted extraction. Because cellulase and pectinase were not used to disrupt the cell walls, the extraction rates of chlorogenic acid, total flavonoids, and total saponins significantly decreased, with contents dropping to 22.0, 29.5, and 12.5 mg / 100 mL, respectively; the taste score also decreased to 32.1. This confirms that enzymatic hydrolysis is a fundamental step for improving extraction rates, and its absence results in approximately a 25% loss of extraction efficiency.

[0130] Comparative Example 9 omitted ultrasound assistance, relying solely on enzymatic hydrolysis followed by thermal extraction. The absence of ultrasound's cavitation effect and mechanical vibration reduced mass transfer efficiency, resulting in lower levels of chlorogenic acid, total flavonoids, and total saponins (23.0, 30.8, and 13.0 mg / 100mL, respectively), and a taste score of 33.0. Compared to Comparative Example 8, enzymatic hydrolysis contributed slightly more than ultrasound, but the synergistic effect of both was optimal, confirming the necessity of ultrasound assistance.

[0131] Comparative Example 10 omitted enzymatic hydrolysis and ultrasound, employing only 80°C water bath extraction. Relying solely on thermal diffusion with intact cell walls, the extraction efficiency was the lowest, with chlorogenic acid, total flavonoids, and total saponins reduced to 20.0, 27.0, and 11.5 mg / 100 mL, respectively, approximately 70% of that in Example 1. Comparative Example 10 demonstrates a synergistic effect between enzymatic hydrolysis and ultrasound, with the combined effect far exceeding that of either method alone.

[0132] Comparative Example 11 combined the water used in the two extractions and added it all at once (total water volume was the same), and extracted continuously for the same amount of time. Due to the gradual increase in solute concentration during extraction, the mass transfer driving force decreased, and the contents of chlorogenic acid, total flavonoids, and total saponins decreased to 24.0, 32.0, and 13.5 mg / 100 mL, respectively. Simultaneously, more water-soluble impurities co-dissolved, leading to a turbidity increase to 0.44 NTU and the appearance of a small amount of precipitation, resulting in a taste score of 33.5. This confirms that gradient extraction (divided into two steps, each using fresh solvent) can significantly improve the dissolution rate of active ingredients and reduce the co-dissolution of impurities.

[0133] Comparative Example 12 did not undergo nitrogen purging before filling and sealing (the headspace of the can was filled with air), and the rest was the same as in Example 1. Residual oxygen inside the can accelerated the oxidation of chlorogenic acid during sterilization and storage. After sterilization, the chlorogenic acid retention rate decreased to 86.5%, the shelf life at room temperature was shortened to 18 months, and the chlorogenic acid content (before sterilization) slightly decreased to 27.5 mg / 100 mL. This indicates that nitrogen purging and sodium vitamin C form a gas-liquid biphase antioxidant protection, and the synergistic effect of both is indispensable.

[0134] Comparative Example 13

[0135] In Comparative Example 13, the contents of chlorogenic acid, total flavonoids and total saponins are only 16.0, 22.0 and 9.0 mg / 100mL respectively, the retention rate of peppermint oil is 18.3%, the turbidity is 2.5 NTU, a small amount of precipitation occurs, the taste score is 20.5, the retention rate of chlorogenic acid after sterilization is 75.0%, and the shelf life is 10 months. This comparative example fully reflects the comprehensive defects of existing commercially available products, and is significantly exceeded in all indicators of the present invention.

[0136] I. Animal Experimental Report on the Throat-Clearing Function of Throat-Clearing Oral Preparations

[0137] 1 Materials and Methods

[0138] 1.1 Samples

[0139] The throat-clearing oral preparation prepared by the method of Example 1 has a light brown to reddish-brown liquid as its content. The recommended dose for human body is 500mL / 60kg BW; the recommended dose for 100-fold concentrated solution is 5mL / 60kg BW per day. It is stored in a cool and dry place in a sealed condition, with a shelf life of 24 months. The test substance is prepared with sterile water for experiments. According to the provisions of *Technical Specifications for Inspection and Evaluation of Health Foods* (2003 edition), the concentrated solution of the sample can be used for the experiment. In the following report, "sample" refers to the 100-fold concentrated solution.

[0140] 1.2 Experimental Animals and Feed

[0141] Ninety-six male SPF SD rats weighing 160-180 g bred by Beijing Vital River Laboratory Animal Technology Co., Ltd. [License No.: SCXK (Jing) 2012-0001] were randomly divided into 8 groups according to body weight, with 12 rats in each group. Cotton ball implantation experiment (48 rats) and toe swelling experiment (48 rats) were carried out respectively; sixty male SPF BALB / C mice weighing 18-22 g bred by Beijing Vital River Laboratory Animal Technology Co., Ltd. [License No.: SCXK (Jing) 2012-0001] were randomly divided into 4 groups according to body weight, with 15 mice in each group, and mouse ear swelling experiment was carried out. The maintenance feed was produced by Beijing Keao Xieli Feed Co., Ltd. [License No.: SCXK (Jing) 2014-0010]. All dose groups were given maintenance feed.

[0142] 1.3 Dosage

[0143] The recommended dose of the throat-clearing oral preparation is 5 mL daily for adults (based on a body weight of 60 kg), equivalent to 0.083 mL / kg BW / day. In rat experiments, 2.5, 5, and 15 times the recommended human dose were established, corresponding to low, medium, and high dose groups of 0.21 mL / kg BW, 0.42 mL / kg BW, and 1.25 mL / kg BW daily, respectively. In mouse experiments, 5, 10, and 30 times the recommended human dose were established, corresponding to low, medium, and high dose groups of 0.42 mL / kg BW, 0.83 mL / kg BW, and 2.50 mL / kg BW daily, respectively. For the preparation of the test substance: high-dose group: 25.0 mL of sample was added to sterile water to a final volume of 100.0 mL; medium-dose group: 8.3 mL of sample was added to sterile water to a final volume of 100.0 mL; low-dose group: 4.2 mL of sample was added to sterile water to a final volume of 100.0 mL. The test substance was prepared with sterile water and administered orally once daily for 30 consecutive days, after which various indicators were measured. The gavage volume for rats was 5 mL / kg BW, with a blank control group (0 mL / kg BW); the gavage volume for mice was 10 mL / kg BW, with a blank control group (0 mL / kg BW). The blank control group used sterile water instead of the test substance, with the same daily gavage volume as the test substance dosage group. All dosage groups were given a maintenance diet.

[0144] 1.4 Instruments and Reagents

[0145] 1.4.1 Instruments

[0146] HZF-B3000 electronic balance (2013002), BS223S electronic balance (2008007), BP211D electronic balance (2004013), toe volume measuring instrument (2012004), constant temperature drying oven, punch, micro sampler.

[0147] 1.4.2 Reagents

[0148] Diethyl ether (20080907), Dextran T-40 (17027002), xylene (20150522).

[0149] 1.5 Experimental Methods

[0150] 1.5.1 Rat cotton ball implantation experiment

[0151] Eight days before the end of the experiment, the hair in the groin area of ​​the rats was removed using a hair removal device. The rats were lightly anesthetized with ether, disinfected with povidone-iodine, and the skin in the groin area of ​​the rats was incised under aseptic conditions. Cotton balls that had been autoclaved, dried, and weighed were inserted, the incisions were sutured, and the test substance was continued to be administered. On the last day of the experiment, one hour after administering the test substance, the rats were euthanized by cervical dislocation. The skin was cut at the original suture site, the cotton balls of granulation tissue were peeled off and removed, and placed in a pre-weighed clean petri dish. The dish was dried in a constant temperature drying oven at 60°C with the lid off for one hour, and then weighed to calculate the net weight of the granulation tissue.

[0152] Net granuloma weight (mg) = weight of granuloma on dried cotton ball - weight of original cotton ball;

[0153] 1.5.2 Rat paw swelling test

[0154] On the last day of the experiment, the test sample was administered once. One hour later, the volume of the right hind toe of each group of rats was measured using a toe volume analyzer and recorded as the 0-hour toe volume. Then, 0.1 mL of 1% Dextran T-40 was injected subcutaneously into the right hind toe of each rat, and the toe volume was measured at 1, 2, 4, and 6 hours. Measurements were taken three times at the same site, and the average value was recorded. The difference between the toe volume measured at different times and the toe volume before the inflammatory agent was applied was used as the swelling value, and the toe swelling rate at each time point was calculated.

[0155] Swelling rate (%) = Swelling value / Pre-inflammatory toe volume × 100%;

[0156] 1.5.3 Mouse ear swelling test

[0157] On the last day of the experiment, 20 μL of xylene was taken and dripped onto the center of the auricle on the outer side of the right ear of the mouse, allowing it to diffuse freely. After 30 minutes, the mouse was euthanized by dislocation of the cervical spine, and both ears were cut off. Ear pieces were punched at the same location on both ears using a 9 mm diameter punch and weighed (weighing was done promptly to avoid moisture loss affecting the experimental results). The difference in weight between the two ears was taken as the auricle swelling value, and the auricle swelling rate was calculated.

[0158] Auricular swelling rate (%) = Auricular swelling value / Weight of control ear × 100%;

[0159] 1.6 Data Processing

[0160] Data processing was performed using SPSS software. Analysis of variance (ANOVA) was employed, but the homogeneity of variance test was performed first, following the standard ANOVA procedure. Once homogeneity of variance was confirmed, the F-value was calculated. <F 0.05 Conclusion: There was no significant difference between the means of each group; F-value ≥ F 0.05 If P ≤ 0.05, perform statistical analysis by pairwise comparison of the means among multiple experimental groups and one control group; perform appropriate variable transformation on non-normal or unequal variance data until the data meets the requirements of normality or homogeneity of variance, and then use the transformed data for statistical analysis; if the transformation still does not achieve the goal of normality or homogeneity of variance, use the rank-sum test for statistical analysis.

[0161] 1.7 Basis for Result Judgment

[0162] A positive result in the rat cotton ball implantation test, along with a positive result in either the rat paw swelling test or the mouse ear swelling test, indicates that the test sample has a positive result in the animal test for throat clearing function.

[0163] 1.7.1 Determination of the results of cotton ball implantation in rats

[0164] Compared with the blank control group, the net content of granulomas in the test group was significantly reduced. The statistical analysis showed that the difference was statistically significant, indicating that the test sample was positive in the rat cotton ball implantation experiment.

[0165] 1.7.2 Determination of rat paw swelling results

[0166] Compared with the blank control group, the toe volume swelling rate was significantly reduced before and after stimulation at any time point. The statistical analysis showed that the difference was statistically significant, indicating that the test sample rat toe swelling test result was positive.

[0167] 1.7.3 Interpretation of Mouse Ear Swelling Results

[0168] Compared with the model control group, the test group showed a significant reduction in ear swelling rate, indicating a statistically significant difference. This suggests that the ear swelling test result of the test sample mice was positive.

[0169] 2 Results

[0170] 2.1 Effects of the throat-clearing oral preparation on the body weight of rats and mice

[0171] Table 2. Rat body weight before and after the cotton ball experiment

[0172]

[0173] Table 3. Rat body weight before and after the paw swelling experiment.

[0174]

[0175] Table 4. Mouse body weight before and after ear swelling experiment

[0176]

[0177] As shown in Tables 2-4, there were no significant differences in body weight between the test substance and the control group in each dosage group before and after administration (P>0.05); that is, the oral preparation for clearing the throat had no adverse effect on the body weight of mice and rats. Some photos of the experimental process are shown below. Figure 1 and Figure 2 As shown.

[0178] 2.2 Effect of throat-clearing oral preparations on the net amount of cotton ball-implanted granulomas in rats

[0179] Table 5 Net amount of granuloma in rat cotton ball implantation experiment

[0180]

[0181] Note: *There was a significant difference compared with the blank control group (0 mL / kg BW).

[0182] As shown in Table 5, after 30 days of oral administration of the throat-clearing oral preparation to rats, the net amount of granulomas in the 1.25 mL / kg BW group was significantly lower than that in the 0 mL / kg BW group (P<0.05). This indicates that the throat-clearing oral preparation can reduce the net amount of granulomas induced by cotton ball implantation in the 1.25 mL / kg BW group.

[0183] 2.3 Effect of throat-clearing oral preparations on paw edema rate in rats

[0184] Table 6. Toe swelling rate in rats 1 hour after injection of Dextran T-40

[0185]

[0186] As shown in Table 6, after 30 days of oral administration of the throat-clearing preparation to rats, there was no significant difference in the paw swelling rate 1 hour after injection of Dextran T-40 in any of the dose groups compared with the 0 mL / kg BW group (P>0.05).

[0187] Table 7. Toe swelling rate in rats 2 hours after injection of Dextran T-40

[0188]

[0189] As shown in Table 7, after 30 days of oral administration of the throat-clearing preparation to rats, there was no significant difference in the paw swelling rate 2 hours after injection of Dextran T-40 in each dose group compared with the 0 mL / kg BW group (P>0.05).

[0190] Table 8. Toe swelling rate in rats 4 hours after injection of Dextran T-40

[0191]

[0192] Note: *There was a significant difference compared with the blank control group (0 mL / kg BW).

[0193] As shown in Table 8, after 30 days of oral administration of the throat-clearing preparation to rats, compared with the 0 mL / kg BW group, the 1.25 mL / kg BW group showed a significantly lower rate of paw swelling 4 hours after injection of Dextran T-40 (P<0.05).

[0194] Table 9. Toe swelling rate in rats 6 hours after injection of Dextran T-40

[0195]

[0196] Note: *There was a significant difference compared with the blank control group (0 mL / kg BW).

[0197] As shown in Table 9, after 30 days of oral administration of the throat-clearing preparation to rats, compared with the 0 mL / kg BW group, the 0.42 mL / kg BW and 1.25 mL / kg BW groups showed a significant decrease in paw swelling rate 6 hours after injection of Dextran T-40 (P<0.05).

[0198] As shown in Tables 6-9, the oral throat-clearing preparations reduced the rate of toe swelling induced by Dextran T-40 injection in the 0.42 mL / kg BW and 1.25 mL / kg BW groups.

[0199] 2.4 Effect of throat-clearing oral preparations on ear swelling rate in mice

[0200] Table 10 Ear swelling rate in mouse ear edema experiment

[0201]

[0202] As shown in Table 10, after 30 days of oral administration of the throat-clearing oral preparation to mice, there was no significant difference in the ear swelling rate between the different dosage groups and the 0 mL / kg BW group (P>0.05). This indicates that the throat-clearing oral preparation had no effect on the ear swelling rate in mice. 3. Conclusion: After 30 days of oral administration of the throat-clearing oral preparation to rats, compared with the 0 mL / kg BW group, the test substance at 0.42 mL / kg BW reduced the paw swelling rate in rats 6 hours after injection of Dextran T-40 (P<0.05); at 1.25 mL / kg BW, it reduced the net granuloma volume in rats (P<0.05) and reduced the paw swelling rate in rats 4 and 6 hours after injection of Dextran T-40 (P<0.05). After 30 days of oral administration of the throat-clearing oral preparation to mice, there was no significant difference in the ear swelling rate between the different dosage groups and the 0 mL / kg BW group (P>0.05). The oral throat-clearing preparation had no effect on ear swelling rate in mice. The test substance had no adverse effect on weight gain in either mice or rats. Based on the criteria for evaluating throat-clearing function, the animal experiment results for the oral throat-clearing preparation showed a positive result.

[0203] II. Human Trial Report on the Throat-Clearing Function of Oral Throat-Clearing Preparations

[0204] 1. Materials and Methods

[0205] 1.1 Sample

[0206] Example 1: A throat-soothing oral preparation prepared according to the method described. Sample characteristics: The contents are a light brown to reddish-brown liquid. Storage method: Sealed and stored in a cool, dry place. Shelf life: 24 months. Recommended dosage: 500 mL / day, twice daily, one vial each time.

[0207] 1.2 Subjects

[0208] 119 participants aged 18-65 with chronic pharyngitis were selected on a voluntary basis.

[0209] 1.2.1 Subject Inclusion Criteria

[0210] 1.2.1.1 Signs: Individuals with chronic pharyngitis may experience subjective symptoms such as sore throat, itchy throat, dry throat, dry cough, foreign body sensation, and increased symptoms with excessive talking.

[0211] 1.2.1.2 Pharyngeal symptoms: edema of the pharyngeal mucosa, mucosal congestion, hyperplasia of lymphoid follicles on the posterior pharyngeal wall, and adhesion of secretions.

[0212] Volunteer subjects who exhibit at least one of the findings in 1.2.1.1 and 1.2.1.2 may be included in the observation.

[0213] 1.2.2 Exclusion criteria for subjects

[0214] 1.2.2.1 Acute exacerbation of chronic pharyngitis or caused by acute pharyngitis, vocal cord nodules, colds, or smoking.

[0215] 1.2.2.2 Caused by lesions of the nasopharynx, pharynx, larynx, nose, esophagus, neck, tuberculosis, or metastatic lung cancer.

[0216] 1.2.2.3 Individuals under 18 years of age or over 65 years of age, pregnant and lactating women, and individuals allergic to the test product.

[0217] 1.2.2.4 Patients with serious diseases of the heart, cerebrovascular system, liver, kidney, hematopoietic system, bronchi and lungs, as well as patients with mental illness and sleep disorders.

[0218] 1.2.2.5 The person who has taken items related to the test function in a short period of time may affect their judgment of the results.

[0219] 1.2.2.6 Those who did not take the test sample as required or who took other medications midway through the treatment, making it impossible to determine the efficacy or whose data is incomplete.

[0220] 1.3 Experimental Design and Grouping Requirements

[0221] Both self-controlled and inter-group controlled designs were employed. Subjects were randomly assigned to the trial group and the control group based on their pharyngeal symptoms and signs. When grouping, key factors influencing the results, such as disease duration, age, and gender, were considered as much as possible, and a balance test was performed to ensure comparability between groups. Each group consisted of at least 50 subjects.

[0222] 1.4 Dosage and timing of consumption

[0223] The test group took the throat-soothing oral preparation as recommended, orally, twice daily, 250 mL each time. The control group served as a blank control. The test substance was taken for 15 days. During the trial, the subjects did not change their living or working environment, nor their original lifestyle or dietary habits.

[0224] 1.5 Instruments and Reagents

[0225] Hitachi 7180 Fully Automated Biochemical Analyzer (CENTRONIC GmbH, Germany); DIRUI H-300 Urine 8-Item Analyzer (Changchun Dirui Pharmaceutical Co., Ltd.); Sysmex-K21 Three-Part Differential Hematology Analyzer (Sysmex Corporation); Sysmex Blood Cell Analysis Diluent (Sysmex Corporation); DIRUI Urine Analysis Reagent Tape (Changchun Dirui Pharmaceutical Co., Ltd.); Leadman Biochemical Reagent Kit (Beijing Leadman Biochemical Co., Ltd.); Kyowa Biochemical Reagent Kit (Kyowa Pharmaceutical Co., Ltd., Japan); Ichika Biochemical Reagent Kit (Daiichi Chemical Co., Ltd., Japan).

[0226] 2. Observation Indicators

[0227] 2.1 Efficacy Indicators

[0228] 2.1.1 Symptom Observation

[0229] Accurately record the subjective pharyngeal symptoms of the subjects before and after the trial. The main pharyngeal symptoms include: sore throat, itchy throat, dry throat, dry cough, and increased foreign body sensation. Calculate the score according to the severity of the symptoms (1 point for grade 1, 2 points for grade 2, and 3 points for grade 3), and statistically analyze the changes in scores and the symptom improvement rate.

[0230] 2.1.2 Observation of physical signs

[0231] Pharyngeal examination: Pharyngeal mucosal congestion, mucosal edema, hyperplasia of lymphoid follicles on the posterior pharyngeal wall, secretions, and other signs. The results were classified into mild, moderate, and severe levels (I, II, III). Changes in signs before and after the trial were recorded, and the sign scores and improvement rates were calculated.

[0232] 2.2 Safety Indicators

[0233] 2.2.1 General condition (including mental state, sleep, diet, bowel movements, blood pressure, etc.).

[0234] 2.2.2 Routine blood, urine, and stool tests

[0235] Red blood cell count, hemoglobin, white blood cell count, 10 urine tests, stool microscopy.

[0236] 2.2.3 Determination of Biochemical Indicators

[0237] Serum albumin (Alb), total protein (TP), liver and kidney function (AST, ALT, urea, creatinine), blood glucose (Glu), and blood lipids (TC, TG).

[0238] 2.2.4 Abdominal ultrasound, electrocardiogram, and chest X-ray (to be performed once before the food trial).

[0239] 2.3 Efficacy Assessment

[0240] 2.3.1 Effective: Symptoms are reduced by 1 degree, and the results of pharyngeal examination are reduced by 1 grade.

[0241] 2.3.2 Ineffective: No significant changes in symptoms or signs.

[0242] 2.4 Data Statistical Analysis

[0243] The integral change can be analyzed using the t-test. For self-comparison data, paired t-tests can be used; for comparisons of means between two groups, independent t-tests are used. The latter requires a homogeneity of variance test. For non-normally distributed or unequally sized data, appropriate variable transformations should be performed until normality and homogeneity of variance are achieved, then the transformed data should be used for the t-test. If the transformed data still do not meet the requirements for normality and homogeneity of variance, a t' test or rank-sum test should be used instead. However, for data with excessively high coefficients of variation (e.g., CV > 50%), the rank-sum test should be applied.

[0244] The improvement rate is count data, which can be expressed as x. 2 If the total number of cases in the 2x2 contingency table is less than 40, or if the total number of cases is equal to or greater than 40 but the theoretical number is equal to or less than 1, the exact probability method should be used instead.

[0245] 2.5 Result Determination

[0246] The test sample showed a significant reduction in the scores of clinical symptoms and signs in the pharynx compared to the control group, and the improvement rate of symptoms and signs was significantly higher than that of the control group. The difference was statistically significant, indicating that the test sample has the function of clearing the throat.

[0247] 3. Experimental Results

[0248] 3.1 Results of efficacy observation

[0249] 3.1.1 General Situation

[0250] A total of 119 subjects were included and randomly divided into a trial group and a control group. Two subjects dropped out of each group, leaving 115 valid subjects (58 in the trial group and 57 in the control group). Pre-trial routine blood, urine, and stool tests, liver and kidney function tests, chest X-ray, electrocardiogram, and ultrasound were performed. All subjects were within the normal range. The grouping is shown in Table 1. Before the trial, there were no significant differences in age, gender, or disease duration between the two groups (P>0.05), making them comparable. There were no significant changes in mental state, sleep, appetite, or bowel movements before and after the trial. Blood pressure and heart rate also showed no significant changes before and after the trial (Table 2).

[0251] Table 11 Comparison of general conditions before observation

[0252]

[0253] Table 12 Changes in heart rate and blood pressure before and after the food trial

[0254]

[0255] 3.1.2 Improvement of main symptoms

[0256] Table 13 Improvement of Main Symptoms

[0257]

[0258] 3.1.3 Improvement status of pharyngeal signs

[0259] Table 14 Improvement of Pharyngeal Signs

[0260]

[0261] Photos of the experimental process are as follows Figure 3 and Figure 4 As shown.

[0262] 3.1.4 Changes in total vital signs score

[0263] Table 15 Changes in Total Vital Signs Score

[0264]

[0265] Note: For comparisons within a group, ***P<0.001; for comparisons between groups, ###P<0.001.

[0266] 3.1.5 Changes in total symptom score

[0267] Table 16 Changes in Total Symptom Score

[0268]

[0269] Note: For comparisons within a group, ***P<0.001; for comparisons between groups, ###P<0.001.

[0270] 3.1.6 Efficacy Assessment

[0271] Table 17 Efficacy Evaluation Results

[0272]

[0273] Note: Intergroup comparison ###P<0.001.

[0274] 3.2 Safety Observation Results

[0275] 3.2.1 Blood routine, urine routine, stool routine and blood biochemical indicators before and after the trial

[0276] Table 18 Changes in routine blood, urine, and stool tests and blood biochemical indicators before and after the trial.

[0277]

[0278] Note: All the above indicators were within the normal range before and after the test.

[0279] 3.2.2 Abdominal ultrasound, electrocardiogram, and chest X-ray were all within the normal range for both the trial group and the control group before the trial.

[0280] 3.2.3 Adverse reaction observation: No allergic reactions or other adverse reactions were observed.

[0281] Table 19 Adverse reactions and allergic reactions in the two groups during the trial period.

[0282]

[0283] 3.3 Dropout rate: 119 subjects were included, with 58 in the trial group and 57 in the control group. Two subjects from each group failed to attend the scheduled follow-up examination, meeting the exclusion criteria. The remaining valid subjects were 58 in the trial group and 57 in the control group, with dropout rates of 3.3% and 3.4%, respectively.

[0284] 4. Summary

[0285] 4.1 119 eligible subjects were randomly divided into a trial group and a control group, with 115 effective subjects (n=58 in the trial group and n=57 in the control group). The control group served as a blank control. The trial group received oral throat-clearing preparation, 250 mL twice daily, for 15 consecutive days without adverse reactions. Before the trial, there were no significant differences in pharyngeal clinical symptoms and signs scores between the trial group and the control group, making the two groups comparable. The trial group showed a significant reduction in pharyngeal clinical symptoms and signs scores before and after the trial (P<0.001), with 25 cases showing improvement, resulting in a total effective rate of 43.1%. The control group showed no significant change in pharyngeal clinical symptoms and signs scores before and after the trial (P>0.05), with only 1 case showing improvement, resulting in a total effective rate of 1.8%.

[0286] III. Verification Experiment on the Improvement of Oral Absorption Rate of Chlorogenic Acid and Total Flavonoids by Black Pepper Extract

[0287] 1. Testing Method

[0288] 1.1 Experimental Materials

[0289] Test sample:

[0290] Example 1: The throat-soothing oral preparation of the present invention containing black pepper extract powder (0.008 g / L);

[0291] Example 2: The throat-soothing oral preparation of the present invention containing black pepper extract powder (0.005 g / L);

[0292] Example 3: The throat-soothing oral preparation of the present invention containing black pepper extract powder (0.010 g / L);

[0293] Comparative Example 3: The throat-soothing oral preparation of the present invention without black pepper extract powder (the other ingredients are exactly the same as those in Example 1).

[0294] Laboratory animals: SPF-grade SD rats, weighing 180-220g, half male and half female, purchased from a qualified laboratory animal center. Housing environment: Temperature 22±2℃, relative humidity 50%-60%, 12h light / 12h dark cycle, free access to food and water.

[0295] 1.2 Grouping and Dosing Regimens of Experimental Animals

[0296] SD rats were randomly divided into 4 groups, with 8 rats in each group (half male and half female):

[0297] Comparative Group 3: Oral preparation without black pepper extract administered by gavage (10 mL / kg body weight).

[0298] Example 1 group: oral preparation containing 0.008 g / L of black pepper extract (10 mL / kg body weight) was administered by gavage.

[0299] Example 2 group: oral preparation containing 0.005 g / L of black pepper extract (10 mL / kg body weight) was administered by gavage.

[0300] Example 3 group: oral preparation containing 0.010 g / L of black pepper extract (10 mL / kg body weight) was administered by gavage.

[0301] The chlorogenic acid content in each group of oral formulations was 28 mg / 100 mL, and the total flavonoid content was 38 mg / 100 mL, ensuring that all variables except black pepper extract were consistent during the comparison.

[0302] 1.3 Drug administration and blood collection

[0303] After fasting for 12 hours (with free access to water), rats were administered the drug orally via gavage according to the grouping protocol. Approximately 0.3 mL of blood was collected via the tail vein before administration (0 h) and at 0.25 h, 0.5 h, 0.75 h, 1 h, 1.5 h, 2 h, 3 h, 4 h, 6 h, and 8 h after administration, and placed in heparinized EP tubes. The whole blood was centrifuged at 4000 rpm for 10 min at 4°C, and the plasma was separated, transferred to clean EP tubes, and stored at -80°C for analysis.

[0304] 1.4 Determination of plasma concentrations of chlorogenic acid and total flavonoids

[0305] Chlorogenic acid determination: Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) was used. 100 μL of plasma was taken, 10 μL of internal standard solution (caffeic acid, 200 ng / mL) was added, and 300 μL of acetonitrile was added to precipitate the protein. The mixture was vortexed for 1 min, centrifuged at 12000 rpm for 10 min at 4 °C, and the supernatant was collected for analysis.

[0306] Chromatographic conditions: ACQUITY UPLC BEH C18 column (2.1×50mm, 1.7μm); mobile phase A was 0.1% formic acid aqueous solution, and mobile phase B was acetonitrile, with gradient elution (0-1min 10%B, 1-4min 10%-90%B, 4-5min 90%B, 5-5.5min 90%-10%B, 5.5-6min 10%B); flow rate 0.3mL / min; column temperature 40℃; injection volume 5μL.

[0307] Mass spectrometry conditions: Electrospray ionization (ESI) source, negative ion mode monitoring; chlorogenic acid quantitative ion pair m / z 353→191, qualitative ion pair m / z 353→85; internal standard caffeic acid m / z 179→135. Source temperature 150℃, desolvation gas temperature 500℃, desolvation gas flow rate 800 L / h.

[0308] Total flavonoids determination: Rutin was used as a marker to quantify total flavonoids. Chromatographic and mass spectrometric conditions were the same as for chlorogenic acid determination; the quantitative ion pair for rutin was 609→300. Standard curve range: chlorogenic acid 5~2000 ng / mL (r 2 >0.995), rutin 10~2000 ng / mL (r 2 >0.995), and the lower limit of quantitation both met the experimental requirements.

[0309] 1.5 Calculation of Pharmacokinetic Parameters

[0310] Using a non-compartmental model, the following pharmacokinetic parameters were calculated using DAS 3.0 software:

[0311] Cmax: Measured maximum blood drug concentration;

[0312] Tmax: Peak time;

[0313] T 1 / 2 Eliminate half-life;

[0314] AUC 0-t Area under the drug-time curve from 0 to t (calculated using the trapezoidal method);

[0315] AUC 0-∞ Area under the curve from 0 to ∞;

[0316] MRT 0-t Average length of stay;

[0317] Relative bioavailability (F): F = (AUC_Example Group / AUC_Comparative Example 3 Group) × 100%.

[0318] 1.6 Data Processing

[0319] Experimental data are expressed as mean ± standard deviation (x̄ ± SD). Statistical analysis was performed using SPSS 26.0 software. One-way ANOVA was used for comparisons among multiple groups, and Dunnett's test was used for pairwise comparisons. A p-value < 0.05 was considered statistically significant.

[0320] 2. Experimental Results

[0321] The results of the determination of the main pharmacokinetic parameters of chlorogenic acid and total flavonoids (calculated as rutin) in the plasma of rats after gavage are shown in Tables 20 and 21.

[0322] Table 20. Main pharmacokinetic parameters of chlorogenic acid in plasma of rats after gavage (n=8, )

[0323]

[0324] Note: * P < 0.01 compared with Comparative Example 3; # P < 0.05 compared with Example 2; No significant difference between Example 1 and Example 3 (P > 0.05).

[0325] Table 21. Main pharmacokinetic parameters of total flavonoids (calculated as rutin) in plasma of rats after gavage (n=8, )

[0326]

[0327] Note: * P < 0.01 compared with Comparative Example 3; # P < 0.05 compared with Example 2; No significant difference between Example 1 and Example 3 (P > 0.05).

[0328] 4. Results Analysis

[0329] 3.1 Enhancement effect of black pepper extract on oral absorption of chlorogenic acid

[0330] Table 1 shows that in the three comparative groups without black pepper extract, the peak concentration (Cmax) of chlorogenic acid was 58.3 ± 6.7 ng / mL, and the area under the curve (AUC) was... 0-8 The concentration of chlorogenic acid was 187.4 ± 18.6 ng·h / mL. When 0.008 g / L of black pepper extract was added to the oral formulation (Example 1), the Cmax of chlorogenic acid increased to 128.6 ± 12.4 ng / mL, and the AUC... 0-8 The concentration was increased to 432.8 ± 35.7 ng·h / mL, and the relative bioavailability (F) reached 231.0%, meaning that the overall oral absorption rate of chlorogenic acid increased by approximately 131% (AUC). 0-8 Compared to Comparative Example 3, this represents an increase of 245.4 ng·h / mL. Simultaneously, the elimination half-life of chlorogenic acid (t...)... 1 / 2 The mean residence time (MRT) increased from 2.0 ± 0.3 h to 3.6 ± 0.4 h. 0-8 The duration of action was extended from 2.8±0.3h to 4.5±0.4h. These results indicate that piperine not only significantly increases the total absorption of chlorogenic acid, but also significantly slows down its elimination rate in the body, thus significantly prolonging the exposure time of the active ingredient at the target site (such as the pharyngeal mucosa), which is beneficial for the sustained exertion of its throat-clearing and anti-inflammatory effects.

[0331] 3.2 Enhancement effect of black pepper extract on oral absorption of total flavonoids

[0332] As shown in Table 2, compared with Comparative Example 3, the Cmax of total flavonoids (calculated as rutin) in Example 1 group increased from 42.5±5.2 ng / mL to 89.7±8.8 ng / mL, and the relative bioavailability (F) was 216.6%, indicating that the overall oral absorption rate of total flavonoids increased by approximately 117% (AUC).0-8 The total flavonoids elimination half-life (t) increased by 182.2 ng·h / mL compared to Comparative Example 3. 1 / 2 The duration of action increased from 1.8±0.3h to 3.2±0.4h, consistent with the trend of chlorogenic acid. The simultaneous increase in total flavonoid absorption rate and in vivo residence time directly enhanced the concentration of anti-inflammatory active substances of flavonoid components in chrysanthemum in the pharyngeal mucosa, thereby amplifying the throat-clearing effect of the entire formula.

[0333] 3.3 Differences in the effects of different doses of black pepper extract

[0334] As shown in Tables 1 and 2, a clear dose-dependent relationship exists among the three groups: Example 2 (0.005 g / L), Example 1 (0.008 g / L), and Example 3 (0.010 g / L). Taking the relative bioavailability (F) of chlorogenic acid as an example: Example 2 was 168.5%, Example 1 was 231.0%, and Example 3 was 239.8%. Example 1 (0.008 g / L) showed a significant improvement over Example 2 (0.005 g / L) (P<0.05), but there was no statistically significant difference between Example 3 (0.010 g / L) and Example 1 (0.008 g / L) (P>0.05), indicating that 0.008 g / L is close to the optimal effective dose.

[0335] 3.4 Mechanism of Action Analysis

[0336] Piperine (the core active ingredient in black pepper extract) enhances the oral absorption of chlorogenic acid and total flavonoids through the following mechanisms:

[0337] (1) Inhibition of glucuronyl transferase (UGT) activity: Piperine can inhibit the activity of UGT enzyme in intestinal epithelial cells, reduce the glucuronide binding metabolism of chlorogenic acid and flavonoid aglycones during absorption, and allow more of the original drug to enter the bloodstream. The significant increase in AUC (131%) in this experiment directly supports this mechanism.

[0338] (2) Inhibition of P-glycoprotein efflux: Piperine can inhibit the drug efflux function of intestinal P-glycoprotein, reducing the amount of absorbed components pumped back into the intestinal lumen, which corresponds to the significant increase in Cmax (approximately 120%) in this experiment.

[0339] (3) Prolonging the residence time of active ingredients in the body: In this experiment, the elimination half-life was extended by about 80%, indicating that piperine can further prolong the elimination process of active ingredients in the body by inhibiting the activity of liver metabolic enzymes (such as CYP3A4).

[0340] 3.5 Summary

[0341] Based on the data in Tables 1 and 2, this invention, by adding a very low dose (0.005~0.01 parts, preferably 0.008 parts) of black pepper extract powder to the formulation, increases the oral bioavailability of chlorogenic acid by approximately 131%, the oral bioavailability of total flavonoids by approximately 117%, and extends the elimination half-life of the active ingredients by 60%~80% without increasing the amount of active ingredients.

[0342] The purified water used in this invention is drinking water that meets the standards of GB 5749-2022 "Standards for Drinking Water Quality" as the raw water, and is prepared through pretreatment, reverse osmosis desalination, disinfection and sterilization, and a circulating distribution system. The specific steps are as follows:

[0343] (1) Raw water preparation: Urban tap water that meets the GB 5749-2022 "Standards for Drinking Water Quality" is used as raw water. Before entering the purified water preparation system, it needs to be stored in the raw water tank, which is equipped with a liquid level control device and a breathing filter.

[0344] (2) Pretreatment: Raw water passes through the following treatment units in sequence: ① Multi-media filter, using quartz sand filter media, to remove suspended solids, silt, and colloidal particles from the water, with a filtration accuracy ≤10μm; ② Activated carbon filter, using columnar activated carbon, to adsorb and remove residual chlorine (controlled ≤0.1mg / L), organic matter, and odor substances from the water, protecting the subsequent reverse osmosis membrane from oxidation; ③ Softener, using cation exchange resin, to remove calcium and magnesium ions from the water, reducing water hardness to ≤0.03mmol / L, and preventing scaling of the reverse osmosis membrane; ④ Security filter, using a 5μm precision filter element, to intercept fine particles remaining after pretreatment. Each filter is equipped with a backwashing or regeneration device and is regularly maintained to ensure treatment effectiveness.

[0345] (3) Reverse osmosis desalination: The pretreated water enters the secondary reverse osmosis unit. ① Primary reverse osmosis: Under an operating pressure of 1.2 MPa, water molecules permeate through the reverse osmosis membrane, while dissolved salts (desalination rate ≥98%), bacteria, organic matter, etc. are retained. The conductivity of the primary product water is ≤10 μS / cm. ② Secondary reverse osmosis: The primary product water enters the secondary reverse osmosis unit after being buffered by an intermediate water tank. Under an operating pressure of 1.0 MPa, residual ions are further removed. The conductivity of the secondary product water is ≤3 μS / cm. The water utilization rates of the two stages of reverse osmosis are 63% and 70%, respectively. The system is equipped with an online conductivity monitoring instrument to monitor the product water quality in real time.

[0346] (4) Storage and Distribution: Secondary reverse osmosis permeate enters the purified water storage tank. The storage tank is made of 316L stainless steel and is designed for sanitary use. A breather with a 0.22μm hydrophobic filter membrane is installed on the top to prevent airborne microorganisms from entering. The distribution system adopts a full circulation conveying method with a circulation velocity ≥1m / s. The pipeline is made of sanitary stainless steel (SUS304 or 316L), and the welding process meets sanitary standards. The pipeline design has no dead corners or blind pipes. The valves are sanitary diaphragm valves with a slope ≥1% to facilitate drainage. 0.22μm microporous membrane terminal filters are installed at the point of use as needed. The system is equipped with a pasteurization device, which is periodically (usually once a week) circulated with hot water at 82±2℃ for 30 minutes to maintain a stable level of microorganisms in the system. Conductivity, total organic carbon (TOC), and temperature are monitored online, and the data is traceable.

[0347] (5) Water quality testing: The purified water prepared must meet the purified water standards of the Chinese Pharmacopoeia (2025 edition): It must be a colorless, clear liquid, odorless and tasteless; conductivity ≤ 5.1 μS / cm (25℃); total organic carbon (TOC) ≤ 0.50 mg / L; total aerobic bacteria count ≤ 100 CFU / mL (sample ≥ 1 mL, cultured on R2A medium for 5 days); non-volatile matter ≤ 1 mg / 100 mL; heavy metals ≤ 0.00001%. When the conductivity test is qualified, batch-by-batch testing of pH, ammonia, nitrite, nitrate, and heavy metals is exempted. For testing of either total organic carbon or easily oxidized substances, the TOC method is preferred. Enterprises must conduct a full-item test of purified water at least once a year. For items that cannot be tested independently, a qualified third-party testing agency should be commissioned to conduct the test.

[0348] (6) Use: The purified water that has passed the test can be directly used in the raw material wetting, extraction, excipient dissolution (including β-cyclodextrin inclusion and black pepper extract, etc.), empty tank cleaning and final volume adjustment of the present invention.

Claims

1. A method for preparing an oral preparation with throat-soothing and moisturizing effects, characterized in that, By weight, the following steps are included: S100, add cellulase and pectinase to a mixture of 10-15 parts honeysuckle, 8-12 parts chrysanthemum, 7-11 parts monk fruit, 6-10 parts scrophularia and 6-10 parts dried plum, moisten with purified water, and carry out enzymatic hydrolysis. S200. Add purified water to the enzymatically hydrolyzed material, perform two ultrasonic extractions, combine the two extraction filtrates, filter, and obtain the extract; add 0.02~0.08 parts of sodium vitamin C to the extract, stir evenly, concentrate under reduced pressure, and concentrate to a relative density of 1.02~1.10 to obtain the concentrate. S300. Extract peppermint oil from 3-6 parts of peppermint using steam distillation. Dissolve 0.8-1.5 parts of β-cyclodextrin in purified water and add the peppermint oil dropwise to prepare a peppermint oil-β-cyclodextrin inclusion complex solution. Add 0.005-0.01 parts of black pepper extract powder, 0.4-1.0 parts of sweetener, 5-15 parts of xylitol, 0.3-0.8 parts of potassium sorbate, and 0.2-0.5 parts of citric acid sequentially to the above peppermint oil-β-cyclodextrin inclusion complex solution and stir until completely dissolved to obtain an excipient solution. S400. Mix the concentrated solution and excipient solution evenly, add the remaining purified water, stir evenly to obtain the preparation solution, and filter the preparation solution in two stages to obtain the oral formulation. The sweetener is composed of ammonium glycyrrhizate and mogroside in a mass ratio of 8-20:1; the total amount of purified water is 1765-1915 parts; the black pepper extract powder is piperine, CAS number 94-62-2; In steps S100 to S400, the ratio of purified water used is 1:18.5~25.5:3.5~4.5:17~23; and in step S200, in the two extraction processes, the ratio of purified water used in the first extraction and the second extraction is 1.3~1.6:

1. In step S300, the method for extracting peppermint oil by steam distillation includes the following: taking the prescribed amount of peppermint raw material, pulverizing it at low temperature, adding cellulase and pectinase, moistening it with water, and enzymatically hydrolyzing it at 35~45℃ for 20~40 minutes; subjecting the enzymatically hydrolyzed material to ultrasonic-assisted treatment for 10~15 minutes; then performing steam distillation for 1.5~3.0 hours, collecting the condensed distillate, allowing it to stand and separate into layers, and collecting the upper peppermint oil layer; The mass ratio of cellulase, pectinase and peppermint raw material is 0.03~0.08:0.01~0.05:1, and the mass ratio of water to peppermint raw material is 8~11:

1.

2. The preparation method according to claim 1, characterized in that, In step S100, the mass ratio of cellulase, pectinase and mixed raw materials is 0.02~0.08:0.01~0.05:1; the enzymatic hydrolysis temperature is 35~42℃ and the enzymatic hydrolysis time is 25~35 minutes.

3. The preparation method according to claim 1, characterized in that, In step S200, purified water is added to the enzymatically hydrolyzed material, and the first extraction is performed under ultrasound at 35~45kHz and 240~350W for 25~35 minutes, followed by heating to 75~82℃ and holding for 0.8~1.2 hours. Add purified water to the filter residue and perform a second extraction under the same conditions using ultrasound for 18-25 minutes, followed by incubation at 75-80℃ for 0.5-1 hour. Filter both extractions through a 180-220 mesh filter. Combine the filtrates from both extractions to obtain the extract.

4. The preparation method according to claim 1, characterized in that, In step S200, sodium vitamin C is added to the extract and concentrated under reduced pressure at 65~72℃ and -0.08~0.02MPa to a relative density of 1.02~1.1 to obtain a concentrated solution.

5. The preparation method according to claim 1, characterized in that, In step S300, peppermint oil is added to the β-cyclodextrin aqueous solution at a rate of 20-40 drops / min, and stirred at 38-42℃ for 25-35 min to obtain a peppermint oil-β-cyclodextrin inclusion complex solution.

6. The preparation method according to claim 1, characterized in that, In step S400, the two-stage filtration includes the following: the prepared solution is filtered sequentially through a 2-10 μm filter bag and a 0.22-0.65 μm microporous membrane to obtain the oral formulation; Before filling and sealing, the oral preparation is purged with nitrogen; after sealing, it is sterilized by autoclaving at 110~120℃ for 28~33 minutes.

7. An oral preparation having the effect of clearing the throat and moisturizing the throat, prepared by the method according to any one of claims 1 to 6.

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