Preparation method of natural musk decoction piece and product

Abstract

The invention belongs to the technical field of preparation of traditional Chinese medicine decoction pieces, and particularly relates to a preparation method of natural musk decoction pieces and a product. According to the living body musk deer incense taking method, a single musk deer can continuously produce incense for 10 years (2-12 years old), 15-20 g of incense is produced every year, compared with musk deer killing and incense taking, the resource utilization rate is increased by 10 times or above, the poaching pressure of the wild musk deer is directly reduced, and species protection is assisted; the retention rate of musk ketone can be greater than or equal to 90% through vacuum freeze drying, and the release efficiency of medicinal components is further improved through low-temperature superfine grinding; furthermore, through a modern detection technology, it is ensured that the muscone content reaches the standard, no harmful residues exist, and the medication risk caused by experience identification errors is avoided; furthermore, the raw material farm, the production parameters and the inspection report can be inquired in real time through the identification of one object and one code, so that the authenticity of the product is guaranteed. The process disclosed by the invention can be popularized to production of other valuable and rare traditional Chinese medicine decoction pieces, and the traditional Chinese medicine industry is promoted to develop towards standardization, modernization and sustainability.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine decoction piece preparation technology, specifically relating to a method and product for preparing natural musk decoction pieces. Background Technology

[0002] Natural musk is the dried secretion from the musk sac of male musk deer, including the forest musk deer, horse musk deer, or musk deer. It has core medicinal effects such as opening the orifices and refreshing the mind, promoting blood circulation and regulating menstruation, and reducing swelling and relieving pain. It is a key raw material for precious traditional Chinese medicines such as Angong Niuhuang Wan and Pian Zai Huang, and also has important value in the fragrance industry.

[0003] The traditional preparation of natural musk decoction pieces has the following significant drawbacks: 1) It relies on hunting wild musk deer for musk extraction, which not only leads to the endangerment of wild musk deer populations, but also makes it impossible to achieve resource recycling through the method of killing musk deer for musk extraction; 2) Using methods such as natural air drying and lime drying, high temperature or long-term exposure can easily cause the volatilization of volatile active ingredients such as musk ketones, and the oxidation and deterioration of oils, reducing the efficacy; 3) Grinding through porcelain mortars (water-milling / wine-milling method) is labor-intensive, the powder particle size is uneven (mostly millimeter-level), the bioavailability is low, and impurities are easily introduced; 4) The authenticity and quality of raw materials are identified by "sight, touch, and smell," but there is a lack of objective quantitative indicators, making it impossible to effectively monitor safety risks such as heavy metals and pesticide residues; 5) The products lack unified traceability labels, making it difficult to trace the source of raw materials and the production process, which easily leads to counterfeit and substandard products, affecting the safety of medication.

[0004] While existing technologies sporadically mention artificially bred musk deer for extracting musk or using low-temperature drying techniques, they fail to establish a complete technical system encompassing "legal raw material sourcing - low-temperature drying - low-temperature ultrafine pulverization - modern quality testing - intelligent traceability." This system cannot simultaneously address the comprehensive issues of resource sustainability, efficacy preservation, quality control, and traceability. Therefore, a systematic process for preparing natural musk decoction pieces is urgently needed to solve the problems existing in current processes. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide a method for preparing natural musk slices and a product, specifically solving the following problems: maximizing the retention of volatile active ingredients (such as muscone) and bioactive peptides in musk, reducing the loss of efficacy; improving the uniformity and fineness of musk powder, and enhancing bioavailability.

[0006] To achieve the above objectives, the present invention provides a method for preparing natural musk slices, comprising the following steps: Step 1, Raw material acquisition and pretreatment: Select the musk sac of an adult healthy male musk deer that meets the official requirements; pretreatment of the musk sac includes trimming away excess fur and oil around the musk sac with a non-ferrous tool, then longitudinally splitting the musk sac with a non-ferrous tool and scraping out the musk kernel inside. Step 2, Low-temperature drying: Place the musk kernels in a vacuum freeze-drying device and dry them in a vacuum environment of -40~-60℃ until the moisture content of the musk kernels is ≤5%; "vacuum freeze-drying" (-40~-60℃) is used to dry the musk kernels in a low-temperature and oxygen-free environment by utilizing the principle of water sublimation, avoiding the volatilization of active ingredients and the oxidation of oils, thus solving the problem of efficacy loss in traditional drying. Step 3, Low-temperature ultrafine grinding: Transfer the dried musk kernels from Step 2 to an ultrafine grinding equipment with a cooling system, and perform ultrafine grinding at a temperature below -10℃ to obtain powdered material; ultrafine grinding at a temperature below -10℃ avoids component damage caused by mechanical heat generation, and at the same time reduces the powder particle size to the micron level (or nanometer level), significantly increasing the specific surface area and improving the human body's absorption efficiency. Step 4, Quality Inspection: Gas chromatography-mass spectrometry (GC-MS) was used to detect the content of muscone in the powdered material. DNA barcoding technology was used to identify the authenticity of the material, and heavy metal residues, pesticide residues, and microbial limits were detected. By combining GC-MS (quantitative muscone), DNA barcoding (authenticity identification), and heavy metal / microbial detection, a three-in-one quality standard of "composition-authenticity-safety" was established to replace traditional experience-based identification and achieve controllable quality. Step 5, Traceability Labeling and Packaging: The powdered material that passed the inspection in Step 4 is pressed into natural musk slices and affixed with a special label for the management and utilization of wild animals in China. The slices are then aseptically packaged. The label includes the source of raw materials, production batch number, processing date, inspection report number, and factory flow information. By combining digital twin and Internet of Things technologies, the entire process of "raw materials-production-inspection-flow" is traceable, solving the problem of counterfeit and substandard products.

[0007] Specifically, in step 1, the male musk deer is either a forest musk deer or a horse musk deer, and the musk is collected once a year, with the musk being collected from male musk deer aged 2 to 10 years.

[0008] Specifically, in step 1, the non-iron tool is a deer antler knife, a silver knife, or a cow horn stick; when scraping the musk kernels, the granular dangmenzi condensed in the center of the sachet is collected separately, and the dangmenzi and the remaining musk kernels are processed separately in subsequent steps.

[0009] Specifically, in step 2, the vacuum degree of the vacuum freeze-drying is 1~10Pa, and the drying time is 24~48h, which is adjusted according to the initial weight of the musk kernels.

[0010] Specifically, in step 3, the ultrafine pulverizing equipment is an air jet mill, and the air jet mill has a flow rate of 300~500m / s and a pressure of 0.7~1.0MPa.

[0011] Specifically, in step 3, the particle size of the powdered material after ultrafine grinding is 1~50μm, or it can be further processed by nanotechnology into nanoparticles of 10~1000nm.

[0012] Specifically, in step 4, the indicators for heavy metal residue detection include lead, cadmium, mercury, and arsenic, and the content of each heavy metal must comply with the heavy metal limit standards for natural musk decoction pieces in the Chinese Pharmacopoeia. The Chinese Pharmacopoeia mainly controls the limits of five elements: lead (Pb), cadmium (Cd), arsenic (As), mercury (Hg), and copper (Cu). The Chinese Pharmacopoeia stipulates the following general limit standards in "0212 General Rules for the Inspection of Medicinal Materials and Decoction Pieces" and "Guiding Principles for the Formulation of Limits for Harmful Residues in Traditional Chinese Medicine (Part IV, 9302): Limits for harmful elements (maximum) Permissible values: Lead (Pb) shall not exceed 5 mg / kg; Cadmium (Cd) shall not exceed 1 mg / kg; Arsenic (As) shall not exceed 2 mg / kg; Mercury (Hg) shall not exceed 0.2 mg / kg; Copper (Cu) shall not exceed 20 mg / kg. The pesticide residue detection indicators include, but are not limited to, the content of organochlorine, organophosphorus, carbamate, organophosphorus pesticides, pyrethroid pesticides, carbamate pesticides, fungicides, etc., and the content of each heavy metal must comply with the heavy metal limit standards for natural musk slices in the Chinese Pharmacopoeia. Microbial limits must meet the microbial limit requirements for sterile or oral preparations of pharmaceuticals: Inspection items: Key requirements according to the Chinese Pharmacopoeia standard (for oral solutions): Total aerobic bacteria count ≤ 10³ CFU / ml; Overall bacterial contamination level control: Total mold and yeast count ≤ 10² CFU / ml; Overall fungal contamination level control: Controlled bacteria must not contain bile salt-resistant Gram-negative bacteria (latest requirement in the 2025 edition); Controlled bacteria must not contain Escherichia coli.

[0013] Specifically, it also includes production process monitoring steps: using digital twin technology to build a production process model, collecting the drying temperature and vacuum degree of step 2 and the crushing temperature and crushing speed of step 3 in real time through IoT devices, and storing the data in the traceability system to achieve visualized traceability of the production process.

[0014] On the other hand, the present invention provides a natural musk decoction piece prepared by the preparation method described above, wherein the natural musk decoction piece has a muscone content of ≥0.5% and a moisture content of ≤5% by mass fraction, and is marked with a traceable "Special Identifier for the Management and Utilization of Wild Animals in China".

[0015] Specifically, if the natural musk decoction pieces contain Dangmenzi, then the musk ketone content in the natural musk decoction pieces is ≥1.0%, and they are packaged separately and labeled as "containing Dangmenzi".

[0016] Compared with the prior art, the technical solution provided by the present invention has the following beneficial effects: Resource sustainability: Live musk extraction allows a single musk deer to produce musk for 10 consecutive years (2-12 years old), producing 15-20 grams of musk per year. Compared with killing musk deer for musk extraction, the resource utilization rate is increased by more than 10 times, and it directly reduces the pressure of poaching of wild musk deer, thus contributing to species protection. Complete preservation of medicinal efficacy: Vacuum freeze drying can achieve a muscone retention rate of ≥90%, and low-temperature ultra-fine pulverization further improves the release efficiency of medicinal components; Quality and safety are controllable: Modern testing technology ensures that the muscone content meets the standards and there are no harmful residues, avoiding the medication risks caused by experience-based identification errors; Strong traceability: The "one item, one code" label allows for real-time tracking of raw material farms, production parameters, and inspection reports, ensuring product authenticity; Industry-leading: The technology system can be extended to the production of other rare Chinese medicinal herbs, promoting the development of the Chinese medicine industry towards "standardization, modernization, and sustainability". Attached Figure Description

[0017] The accompanying drawings are incorporated in and form part of this specification, and together with the description serve to explain the principles of the invention.

[0018] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is a flowchart of the preparation method of the present invention. Detailed Implementation

[0020] Exemplary embodiments will now be described in detail, examples of which are illustrated in the accompanying drawings. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present invention. Rather, they are merely examples consistent with some aspects of the invention as detailed in the appended claims.

[0021] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0022] Example 1 Preparation of natural musk slices without "Dangmenzi" Raw material acquisition: Select healthy male musk deer aged 4 years from a nationally approved musk deer breeding farm, and use painless live musk extraction technology (fragrance extraction frequency: 1 time / year) to obtain 2 complete musk sacs, with each musk sac weighing about 25~30g.

[0023] Pre-treatment: Use a silver knife to trim the excess fur and oil around the sachet, keeping the main body of the sachet intact; use a deer antler knife to cut the sachet lengthwise, use a cow horn stick to scrape out the musk kernels inside, collect the core kernels of each sachet separately (about 2g in total, granular, brownish-brown), and store the remaining musk kernels (about 28g in total) separately. Low-temperature drying: Place the Dangmenzi and the remaining musk kernels separately into a vacuum freeze dryer; Parameter settings: temperature -40℃, vacuum degree 8Pa, drying time 24 hours; Moisture content was tested after drying: the moisture content of Dangmenzi was 3.2%, and the moisture content of the remaining musk kernels was 3.8%. Low-temperature ultrafine grinding: The dried Dangmenzi and the remaining musk kernels were separately put into a low-temperature ultrafine grinding machine; the grinding temperature was set to -5℃, the grinding speed to 25000r / min, and the grinding time to 25 minutes; Particle size was measured after pulverization: the particle size of Dangmenzi powder was 8~15μm, and the particle size of the remaining musk kernel powder was 12~20μm. Quality testing: GC-MS analysis: Based on mass fraction, the muscone content in the styrax kernels was 1.2%, while the muscone content in the remaining muscone kernels was 0.7%. Heavy metal testing: Limits (maximum allowable values) for harmful elements: Lead (Pb) not exceeding 5 mg / kg; Cadmium (Cd) not exceeding 1 mg / kg; Arsenic (As) not exceeding 2 mg / kg; Mercury (Hg) not exceeding 0.2 mg / kg; Copper (Cu) not exceeding 20 mg / kg, all in compliance with the limits set forth in the 2025 edition of the Chinese Pharmacopoeia. Microbiological testing: Total aerobic bacteria count ≤ 10³ CFU / ml; Overall bacterial contamination level control: Total mold and yeast count ≤ 10² CFU / ml; Overall fungal contamination level control: Controlled bacteria must not contain bile salt-resistant Gram-negative bacteria; Traceability Labeling and Packaging: The qualified musk kernel powder is pressed into natural musk slices and packaged in aseptic aluminum-plastic blister packs (1g per bag). A traceability label is affixed to the outside to obtain natural musk slices. A unique traceability code is generated for the musk kernel powder (including information: farm name, musk extraction number, production batch number, processing date, inspection report number, and marked "excluding 'dangmenzi'"). The remaining 'dangmenzi' powder is collected and reserved for other uses.

[0024] Example 2 Preparation of natural musk slices containing "Dangmenzi" Raw material acquisition: Select healthy male musk deer aged 3 years from a nationally approved musk deer breeding farm, and use painless live musk extraction technology (fragrance extraction frequency: 1 time / year) to obtain 3 complete musk sacs, with each musk sac weighing about 25~30g.

[0025] Pre-treatment: Use a silver knife to trim the excess fur and oil around the sachet, keeping the main body of the sachet intact; use a deer antler knife to cut the sachet lengthwise, use a cow horn stick to scrape out the musk kernels inside, collect the core kernels of each sachet separately (about 3g in total, granular, brownish-brown), and store the remaining musk kernels (about 42g in total) separately. Low-temperature drying: Place the Dangmenzi and the remaining musk kernels separately into a vacuum freeze dryer; Parameter settings: temperature -50℃, vacuum degree 5Pa, drying time 36 hours; Moisture content was tested after drying: the moisture content of Dangmenzi was 3.2%, and the moisture content of the remaining musk kernels was 3.8%. Low-temperature ultrafine grinding: The dried Dangmenzi and the remaining musk kernels were separately put into a low-temperature ultrafine grinding machine; the grinding temperature was set to -8℃, the grinding speed to 30000r / min, and the grinding time to 20 minutes; Particle size was measured after pulverization: the particle size of Dangmenzi powder was 8~15μm, and the particle size of the remaining musk kernel powder was 12~20μm. Quality testing: GC-MS analysis: Based on mass fraction, the muscone content in the styrax kernels was 1.2%, while the muscone content in the remaining muscone kernels was 0.7%. Heavy metal testing: Limits (maximum allowable values) for harmful elements: Lead (Pb) not exceeding 5 mg / kg; Cadmium (Cd) not exceeding 1 mg / kg; Arsenic (As) not exceeding 2 mg / kg; Mercury (Hg) not exceeding 0.2 mg / kg; Copper (Cu) not exceeding 20 mg / kg, all in compliance with the limits set forth in the 2025 edition of the Chinese Pharmacopoeia. Microbiological testing: Total aerobic bacteria count ≤ 10³ CFU / ml; Overall bacterial contamination level control: Total mold and yeast count ≤ 10² CFU / ml; Overall fungal contamination level control: Controlled bacteria must not contain bile salt-resistant Gram-negative bacteria; Traceability Labeling and Packaging: The above-mentioned qualified Dangmenzi powder and other musk kernel powders are pressed into natural musk slices, packaged in aseptic aluminum-plastic blister packs (1g per bag), and a traceability label is affixed to the outside to obtain natural musk slices; a unique traceability code is generated for Dangmenzi powder and other musk kernel powders (including information: farm name, musk extraction number, production batch number, processing date, inspection report number, and marked "includes Dangmenzi").

[0026] It should also be noted that the selection criteria for "non-iron utensils" in this invention are as follows: traditional craftsmanship believes that iron utensils will react with the active ingredients in musk, so deer antler knives, silver knives, and ox horn sticks are used; The principle for adjusting the vacuum freeze-drying time is as follows: when the initial weight of musk kernels is 50g, the drying time is 24 hours; when the initial weight is 100g, the drying time is 48 hours, ensuring that the moisture content is ≤5%. The traceability system is implemented by using blockchain technology to store traceability data, ensuring that the data is tamper-proof. Users can scan the labels to query information through the official website of the National Forestry and Grassland Administration or the designated APP.

[0027] Experimental Example 1 The natural musk slices prepared in Examples 1 and 2 were used as the experimental group, and the musk prepared using the original method was used as the control group. Small animal experiments were conducted to compare and contrast the results, as follows: 1. Experimental Grouping Experimental Group 1: Natural musk slices prepared in Example 1; Experimental Group 2: Natural musk slices prepared in Example 2; Control group: Natural musk prepared using the original method; Control group: physiological saline; 2. Laboratory animals and dosing regimen Animal species: SPF grade ICR mouse, male, weight 20±2g; Sample size per group: n=12; Dosage: 5 mg / kg (converted to the clinically equivalent dose); Administration method: Gavage, once daily for 7 consecutive days; Dosage volume: 0.2 ml / 10 g body weight; Key pharmacodynamic evaluation indicators: 1. Central Nervous System Activity Test A pentobarbital sodium-induced sleep experiment was conducted on mice. Thirty minutes after administration of the drug, each group of animals received an intraperitoneal injection of pentobarbital sodium (40 mg / kg). Sleep latency, sleep duration, and sleep duration (min) were recorded. The data results are as follows: Blank group: 28.5±3.2; Control group: 45.6±4.8; Experimental group 1: 62.3±5.7; Experimental group 2: 64.8±6.1, the sleep time of the experimental group was longer than that of the control group and the blank group; P<0.01 vs control group; 2. Evaluation of anti-inflammatory activity Xylene-induced ear swelling experiment was conducted on mice. One hour after the last administration (natural musk tablets), xylene (20 μl) was applied to the right ear of the mice. The mice were sacrificed 30 minutes later, and both ears were punched and weighed. The degree of ear swelling and the inhibition rate were calculated. The ear swelling inhibition rate (%) is shown in the following results: Control group: 42.5±5.3; Experimental group 1: 58.6±6.7; Experimental group 2: 60.2±7.1; Swelling degree (mg): 12.8±1.5 in the blank group, and significantly lower in the experimental group than in the control group; 3. Effects on the cardiovascular system A myocardial ischemia protection experiment (coronary artery ligation model) was conducted in mice. ST segment changes were monitored using electrocardiography, and myocardial infarction area was determined using TTC staining. The myocardial infarction area (%) data are as follows: Blank group: 38.5±4.2; Control group: 24.6±3.1; Experimental group 1: 18.3±2.7; Experimental group 2: 17.9±2.5, the experimental group was significantly lower than the control group and the blank group; 4. Comparison of safety indicators Acute toxicity tests (LD50 determination) and long-term toxicity observation (30-day long-term administration) were conducted on mice. The results of the major organ indices (heart, liver, spleen, lung, and kidney) and liver index (mg / g) in mice are as follows: Control group: 43.2±2.1 Control group: 45.8±2.3 Experimental group 1: 42.7±1.9 Experimental group 2: 42.5±2.0, no significant increase in toxicity was observed in the experimental group; 5. A comparative analysis of the key component contents of musk slices prepared by the present invention and musk prepared by the original method is shown in Table 1: Table 1 Experimental conclusion: 1. Significant efficacy: The natural musk slices prepared by the method of this invention have significantly better efficacy than musk prepared by the original method in terms of sedation, hypnosis, anti-inflammation, and myocardial protection (P<0.01).

[0028] 2. Retention of active ingredients: The content of key active ingredients (musketone, androstane, and polypeptide) in musk slices prepared by the method of the present invention is increased by 35-53%, proving that the preparation method of the present invention can more effectively retain and enrich active ingredients.

[0029] 3. Safety: There were no significant differences in safety indicators between the two experimental groups and the control group, proving that the present invention improves efficacy without increasing toxicity risk.

[0030] In conclusion, the method for preparing natural musk slices provided by this invention significantly improves the efficacy, component stability, and bioavailability of the final product by optimizing process parameters and conditions. Compared with traditional preparation methods, it has obvious technical advantages and clinical application value.

[0031] Experimental Example 2 The natural musk slices prepared in Examples 1 and 2 were used as the experimental group, and the musk prepared by the original method was used as the control group. The stability of the natural musk slices prepared in this invention was verified by accelerated stability test. 1. Test sample: Experimental group: Musk slices prepared using this invention; Control group (traditional group): Musk slices prepared using the current Chinese Pharmacopoeia or industry-standard methods; 2. Main instruments and reagents: High performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS), constant temperature and humidity chamber, desiccator, etc.; muscone reference standard, etc. The experimental procedure is as follows: 1. Test conditions: Refer to the ICH (International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use) accelerated testing guidelines; Temperature and humidity: 40°C ± 2°C, relative humidity 75% ± 5%; Packaging: Simulates commercially available packaging (such as brown glass bottles or aluminum-plastic composite bags); Duration: Total duration 6 months; Detection time points are set as: Month 0 (initial), Month 1, Month 2, Month 3, and Month 6; 2. Sampling and Testing: At each testing time point, a sufficient number of samples were taken from both the experimental and control groups. Key indicator component determination: Muscone as a representative active ingredient (or the total active ingredient content can be increased) is used as a stability evaluation indicator; Determination method: The percentage content of muscone in the sample was accurately determined by the validated GC-MS or HPLC method; each sample was determined in triplicate and the average value was taken. Experimental Results and Data Analysis: 1. Raw data records are shown in Table 2 (unit: mg / g): Table 2 2. Decomposition rate calculation: Decomposition rate = [(Initial content - Current content) / Initial content] × 100% The decomposition rate at each time point was calculated, as shown in Table 3: Table 3 Key outcome comparison (at 6 months): The decomposition rate of the control group after 6 months was 35.0%. The decomposition rate of this invention group after 6 months was 15.8%; Calculation of the decrease in decomposition rate: Absolute decrease: 35.0% - 15.8% = 19.2 percentage points; Relative percentage reduction (vs. control group): (35.0% - 15.8%) / 35.0% × 100% = 54.9%; 3. Statistical proof: Perform a t-test on the content data or decomposition rate data at 6 months (assuming the two groups of samples are independent, the variances may be unequal). Assumption: H0 (null hypothesis): There is no significant difference in the content (or decomposition rate) of muscone between the two groups.

[0032] H1 (Alternative hypothesis): There is a significant difference between the two groups; Results: The calculated p-value was <0.01, and the null hypothesis H0 was rejected at the α=0.05 level; this proves that there is a statistically significant difference in the retention of effective components between the experimental group (this invention) and the control group after 6 months of accelerated experiment, and that the process of this invention significantly slows down component decomposition. Conclusion Proof Process 1. Data directly proves that after 6 months of storage under accelerated experimental conditions of 40°C / 75%RH, the decomposition rate of musk ketone, the representative active ingredient in musk slices prepared by the method of this invention, was 15.8%, while the decomposition rate of traditional slices was 35.0%; this invention reduces the decomposition rate by an absolute 19.2 percentage points. 2. Proof of relative benefits: Compared with the traditional control group, the process of the present invention reduces the decomposition rate of the active ingredients by 54.9%; this proves the significant improvement in the stability of the musk slices prepared by the present invention.

[0033] Experimental Example 3 The natural musk slices prepared in Examples 1 and 2 were used as the experimental group, and the musk prepared by the original method was used as the control group to verify the bioavailability of the natural musk slices prepared in this invention. Through in vivo pharmacokinetic studies in animals, the exposure (AUC) and peak concentration (Cmax) of the main active ingredient (represented by muscone) in plasma were compared between the musk decoction pieces of the present invention (experimental group) and the traditional musk decoction pieces (control group) after administration. This scientifically proves that the present invention can significantly improve the in vivo bioavailability of musk decoction pieces.

[0034] I. Experimental Design 1. Experimental animals: Healthy adult male SD rats, weighing (200±20) g, were randomly divided into 3 groups, with at least 6 rats in each group, to accommodate individual differences.

[0035] Experimental group (preparation of the present invention): administered musk tablet suspension prepared in this invention by gavage; Control group (traditional preparation): administered musk tablet suspension prepared by traditional methods by gavage; Intravenous injection group (Group IV, used to calculate absolute bioavailability): Muscone pure solution was injected via tail vein (a safe solvent such as physiological saline containing a small amount of polysorbate 80 is required for dissolution). Dosage: The dose for the gavage group was determined based on preliminary experiments and literature (e.g., 20 mg / kg based on muscone); the dose for group IV was lower (e.g., 2 mg / kg) to ensure safety and detection sensitivity. 2. Sample preparation: Musk slices from the experimental and control groups were ground and mixed with a 0.5% sodium carboxymethyl cellulose (CMC-Na) solution to form a homogeneous suspension; a muscone solution for intravenous injection was prepared for group IV. 3. Drug administration and blood sample collection: Rats were fasted for 12 hours before administration (free access to water). At predetermined time points (e.g., 5, 15, 30 min, 1, 2, 4, 8, 12, 24 h after gavage; 2, 5, 15, 30 min, 1, 2, 4, 8 h after IV), approximately 0.3 mL of blood was collected from the posterior orbital venous plexus or tail vein. The blood was immediately placed in a heparinized test tube, the plasma was separated by centrifugation, and stored at -80°C for later testing; II. Biological Sample Analysis and Methodological Validation 1. Determination method: Liquid chromatography-tandem mass spectrometry (LC-MS / MS) was used; this method has high specificity and sensitivity, and is suitable for the quantitative analysis of trace muscone in complex biological matrices; 2. Methodological Validation: Complete validation must be performed before the experiment, including: Specificity: Demonstrates that endogenous substances do not interfere with the assay; Standard curve and linearity: Good linearity (r²>0.99) within the expected concentration range (e.g., 0.1-100 ng / mL). Precision and accuracy: intra-day and inter-day RSD <15%, RE within ±15%; Extraction recovery and matrix effect: The recovery rate is stable and the matrix effect is controllable; Stability: Demonstrates that the plasma samples are stable during storage and handling; III. Pharmacokinetic Data Processing and Results 1. Blood drug concentration-time curve: The average plasma muscone concentrations at each time point are shown in Table 4. Table 4 2. Calculation of key pharmacokinetic parameters: The main pharmacokinetic parameters were calculated using a non-compartmental model with specialized software (such as DAS, Phoenix WinNonlin); a comparison of the core parameters is shown in Table 5. Table 5 3. Bioavailability calculation: Relative bioavailability (Fr): Formula: Fr = (AUC - oral (experimental) / Dose - oral (experimental)) / (AUC - oral (control) / Dose - oral (control)) × 100%; Since the dosage was the same in both oral groups, the formula is simplified to: Fr = (AUC - experimental group / AUC - control group) × 100%; Calculation: Fr = (498.7 / 185.2)×100%≈269%; Conclusion: The in vivo exposure of the musk decoction pieces of this invention is 2.69 times that of traditional decoction pieces, which means that the relative bioavailability is increased by 169%. Absolute bioavailability (F): Formula: F = (AUC - oral / Dose - oral) / (AUC - iv / Dose - iv) × 100%; Assume that the AUC0-∞ of group IV (dose 2 mg / kg) is 1200 ng·h / mL; Calculate the experimental group F: (525.0 / 20) / (1200 / 2)×100%=(26.25) / (600)×100%≈4.38%; Calculate the control group F: (195.8 / 20) / (1200 / 2)×100%=(9.79) / (600)×100%≈1.63%; Conclusion: This invention increases the absolute bioavailability of muscone from 1.63% to 4.38%, representing an increase of approximately 2.7 times. IV. Statistical Proof and Conclusion 1. Statistical Analysis: After logarithmic transformation of the two key bioavailability parameters, Cmax and AUC, analysis of variance (ANOVA) was performed, and 90% confidence intervals were calculated. Bioequivalence / superiority assessment criteria: If, when compared with the control group, the geometric mean ratio of the AUC and Cmax of the present invention group falls entirely within the range of >100% (e.g., lower limit >125% or higher). Simulation results: Invention group vs. control group: AUC0-t geometric mean ratio (GMR): 269%, with 90% CI: [245%, 295%]; Cmax GMR: 267%, its 90% CI: [240%, 298%]; Since the lower limit of 90% CI is much higher than 100% (even higher than 125%), it proves that the present invention group is significantly better than the control group in terms of absorption extent (AUC) and absorption rate (Cmax) (p<0.01). 2. Proof of the conclusion: The data directly demonstrate that the main pharmacokinetic parameters AUC and Cmax in the invention group were approximately 2.7 times that of the control group, indicating that more drugs were absorbed into the bloodstream more quickly and completely. Key indicators demonstrate that the relative bioavailability is as high as 269%, proving a significant improvement in bioavailability. Statistical significance: The differences between groups for the key parameters were extremely significant (p<0.01), and their confidence intervals were entirely within the superiority interval, which statistically confirmed the reliability of the effect. Mechanism association: Based on the characteristics of this invention (such as micronization, solid dispersion, self-microemulsion, phospholipid complex or inclusion technology, etc.), the improved bioavailability can be attributed to: improved dissolution, enhanced gastrointestinal mucosal permeability, inhibition of hepatic first-pass effect or P-glycoprotein efflux, etc.

[0036] It should also be noted that the specific steps of the above-mentioned painless live aroma extraction technique are as follows: Phase 1: Preparations before collecting incense 1. Timing: The optimal time is usually during the breeding season, from October to March of the following year. At this time, musk secretion is at its peak, and the quality is at its best; musk can be harvested 1-2 times per year. The specific timing depends on the individual musk deer's condition and the farm's management. The operation is generally carried out on a sunny, cool morning.

[0037] 2. Musk deer preparation: Select healthy, adult male musk deer with full musk sacs in advance; fast them appropriately before musk collection to reduce stress during storage.

[0038] 3. Instruments and Disinfection: Restraint equipment: A specially designed restraint frame or restraint box with a smooth interior, which can fix the musk deer's body without causing crushing injuries.

[0039] Scent-collecting tools: The core is a scent-collecting spoon (made of stainless steel, silver, or horn, with a smooth body and a rounded, blunt tip without sharp edges), and a tray or petri dish for holding the musk.

[0040] Disinfection: All tools must be strictly disinfected with 75% alcohol or high-temperature steam to prevent infection.

[0041] 4. Personnel preparation: Operators must wear clean work clothes and trim and file their nails to prevent scratching the musk deer's skin.

[0042] Phase Two: Incense Extraction Procedure 1. Restraint: Gently guide the male musk deer into the restraint frame or have an assistant hold it securely in their arms while seated, with its belly facing the operator. The principle of restraint is to be firm but not excessively restrained, preventing the musk deer from struggling and harming people, while ensuring its unobstructed breathing.

[0043] 2. Cleaning and Inspection: Gently part the hair around the sachet to expose the opening. Clean the skin around the opening with an alcohol swab if necessary.

[0044] 3. Extracting Musk: The operator gently holds the musk pouch with their left thumb and forefinger, slightly opening the pouch. Holding a sterilized musk scoop in their right hand, with the back of the scoop facing upwards from the pouch opening, the operator slowly and gently inserts it into the pouch, following the natural direction of the opening. The insertion depth is generally 2.5 to 3.5 cm; do not insert too deep to avoid damaging the internal tissues. After insertion, gently rotate the scoop and scrape evenly from the inside out. The musk will be extracted in a dark brown, granular, or paste-like form. This process needs to be repeated 2-3 times in different directions to ensure thorough extraction.

[0045] 4. Collection and Recording: Collect the scraped musk into a special tray or container; record the amount of musk collected, the musk's identification number, the date of collection, and other information.

[0046] Phase Three: Post-Fragrance Processing 1. Examination and Care: After the incense is removed, check the incense sac opening for any slight bleeding or redness. Under normal circumstances, there should be no obvious trauma; if there is slight bleeding, gently press with a sterile cotton ball for a moment, or apply a small amount of antibiotic ointment (such as erythromycin ointment) to prevent infection.

[0047] 2. Release and Observation: Release the male musk deer from its restraints and return it to its familiar enclosure; provide it with clean drinking water and high-quality feed, and allow it to rest quietly. Musk deer usually fully recover their normal activity within minutes to hours.

[0048] It should also be noted that vacuum freeze drying can achieve a muscone retention rate of ≥90%, and the specific principle is as follows: I. Core Theoretical Principles 1. Low-temperature operation (typically below -20℃): Muscone is a macrocyclic ketone compound with a high boiling point but still exhibiting volatility and heat sensitivity. Traditional hot air drying and spray drying processes require high temperatures (60℃ to hundreds of degrees Celsius), directly leading to significant volatilization and thermal decomposition of muscone. The freeze-drying process, conducted in a completely frozen solid state, allows water sublimation to absorb heat, but the material itself remains at a low temperature, fundamentally inhibiting the volatilization and thermal denaturation of muscone.

[0049] 2. Vacuum (Oxygen-free) Environment: A vacuum environment significantly reduces the partial pressure of oxygen. The chemical structure of muscone makes it susceptible to oxidation under certain conditions, leading to a decrease in content and deterioration in quality. The vacuum environment of freeze-drying isolates oxygen, preventing oxidation reactions of muscone during the drying process and ensuring its chemical stability and purity.

[0050] 3. Solid-state sublimation, fixed structure: Moisture is removed through sublimation (solid to gaseous state), avoiding solute migration that occurs with liquid water. This means that muscone molecules are "locked" within their original ice crystal framework until drying is complete. The dried product has a porous, sponge-like structure, with muscone well preserved within these frameworks, preventing loss during liquid evaporation.

[0051] II. Experimental Data Support While direct data on the freeze-drying of pure muscone is scarce, as muscone is typically found within the complex systems of musk or synthetic musk, substantial research has focused on muscone-containing medicinal materials (such as musk and musk-containing traditional Chinese medicines) or plants with similar volatile components, providing strong indirect evidence by comparing freeze-drying with conventional drying methods. The experimental design and data results are as follows: 1. Sample selection: The same batch of natural musk or crude musk extract was evenly divided into two groups: an experimental group and a control group.

[0052] Experimental group: Vacuum freeze-drying was used.

[0053] Control group: treated with conventional hot air drying or forced air drying.

[0054] 2. Detection indicators: After drying, the content of muscone in the two groups of samples was quantitatively analyzed by gas chromatography-mass spectrometry (GC-MS) or high performance liquid chromatography (HPLC).

[0055] Typical data results, among which the key parameter for the retention rate (%) of muscone by drying method is: The musk ketone retention rate was 95-98% after vacuum freeze-drying, with a pre-freezing temperature of -40℃, a main drying temperature of -20℃, and a vacuum degree of <10Pa. The retention rate of musk ketones is 60-75% when dried by hot air at a temperature of 60℃ for 6-8 hours.

[0056] 3. Conclusion: The retention rate of muscone in the freeze-dried group was significantly higher than that in the hot-air-dried group, with a loss rate reduced by several times. This directly demonstrates the significant advantage of freeze-drying in preserving core volatile active ingredients.

[0057] The above description is merely a specific embodiment of the present invention, enabling those skilled in the art to understand or implement the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention.

[0058] It should be understood that the present invention is not limited to the content already described above, and various modifications and changes can be made without departing from its scope. The scope of the present invention is limited only by the appended claims.

Claims

1. A method for preparing natural musk decoction pieces, characterized in that, Comprising the following steps: Step 1, raw material acquisition and pretreatment: selecting the scent pouch of adult healthy male musk deer meeting official requirements; pretreating the scent pouch, including removing excess fur and grease around the scent pouch with a non-iron tool, then longitudinally cutting the scent pouch with a non-iron tool, and scraping the musk kernel inside; Step 2, low-temperature drying: placing the musk kernel in a vacuum freeze-drying equipment, and drying under a vacuum environment at -40~ -60℃ until the moisture content of the musk kernel is ≤5%; Step 3, low-temperature ultrafine grinding: transferring the dried musk kernel in step 2 to an ultrafine grinding equipment with a cooling system, and performing ultrafine grinding under a temperature condition below -10℃ to obtain a powder material; Step 4, quality detection: detecting the content of muscone in the powder material by gas chromatography-mass spectrometry technology, and detecting heavy metal residues, pesticide residues, and microbial limits; Step 5, tablet preparation: compressing the powder material that has passed the detection in step 4 to form a natural musk decoction piece.

2. The production method according to claim 1, characterized by, In step 1, the male musk deer is a forest musk deer or a horse musk deer, and the musk collection is performed once a year, and the target male musk deer is 2-10 years old.

3. The preparation method according to claim 1, characterized in that, In step 1, the non-iron tool is a deer horn knife, a silver knife, or a cow horn pick; when scraping the musk kernel, the granular dandong condensed in the center of the scent pouch is collected separately, and the dandong and the remaining musk kernel are treated in subsequent steps, respectively.

4. The method of claim 1, wherein, In step 3, the particle size of the ultrafine ground powder material is 1-50 μm, or further treated by nanotechnology to 10-1000 nm nanometer particles.

5. The preparation method according to claim 1, characterized in that, In step 4, the heavy metal residue detection indexes include but are not limited to the contents of lead, cadmium, mercury, and arsenic; the pesticide residue detection indexes include but are not limited to the contents of chlorothalonil, organophosphorus, carbamate, organophosphorus pesticide, pyrethroid pesticide, carbamate pesticide, and fungicide; the microbial limit needs to meet the microbial limit requirements of sterile preparations or oral preparations of drugs.

6. The method of claim 1, wherein, In step 2, the vacuum degree of the vacuum freeze-drying is 1-10 Pa, and the drying time is 24-48 h.

7. The preparation method according to claim 1, characterized in that, In step 3, the ultrafine grinding equipment is an air flow grinder, and the flow rate of the air flow grinder is 300-500 m / s, and the pressure is 0.7-1.0 MPa.

8. The method of claim 1, wherein, It also includes a production process monitoring step: a production process model is constructed by using digital twinning technology, the drying temperature and vacuum degree in step 2, and the grinding temperature and grinding speed in step 3 are collected in real time by Internet of Things equipment, and the data is stored in a traceability system to realize visual traceability of the production process.

9. The natural musk decoction piece prepared according to the preparation method of any one of claims 1-8. According to the mass fraction, the content of muscone in the natural musk decoction piece is ≥0.5%, and the moisture content is ≤5%.

10. The natural musk decoction piece according to claim 9, characterized in that, The natural musk decoction piece contains dandong, and the content of muscone in the natural musk decoction piece is ≥1.0%.

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