New process for preparing guetengtongling tincture

Abstract

The application belongs to the technical field of Goutongling tincture, and provides a new process of Goutongling tincture. The new process comprises the following steps: (1) ultrahigh pressure extraction: adding 45-55% ethanol to Xueshangziziba Artemisia to perform ultrahigh pressure treatment, and filtering to obtain an extract and dregs; (2) grinding dry ginger, myrrh and frankincense into coarse powder, mixing the coarse powder with the dregs, then adding 50% ethanol for percolation, collecting the percolate, and combining the percolate with the extract; additionally, dissolving dragon's blood and borneol in ethanol, combining the ethanol solution with the percolate and the extract, mixing, standing, and filtering, and thus the Goutongling tincture is obtained. The Goutongling tincture prepared by the new process can obviously increase the content of effective components in the final product, thereby significantly improving the treatment effect and shortening the treatment cycle.

Description

Technical Field

[0001] This invention belongs to the field of bone pain relief tincture preparation technology, specifically, it relates to a new process for bone pain relief tincture. Background Technology

[0002] Bone Pain Relief Tincture is a topical traditional Chinese medicine. Its main ingredients include Artemisia argyi, dried ginger, dragon's blood, frankincense, myrrh, and borneol. The entire formula of Bone Pain Relief Tincture has the effects of warming the meridians and dispelling cold, dispelling wind and promoting blood circulation, and relieving pain. It can be used to treat lumbar and cervical spondylosis, osteoarthritis, frozen shoulder, and rheumatoid arthritis.

[0003] According to the "Drug Standards of the Ministry of Health, Volume 19, Traditional Chinese Medicine Compound Preparations", page number Z19-142, standard number WS3-B-3645-98, the prescription for Gutongling Tincture is as follows: 80g of *Saussurea involucrata*, 110g of dried ginger, 1.0g of dragon's blood, 5.0g of frankincense, 5.0g of myrrh, and 1.5g of borneol. The preparation method is as follows: The above six ingredients are ground into coarse powder, *Saussurea involucrata*, dried ginger, myrrh, and frankincense, mixed evenly, and percolated using a suitable amount of 50% ethanol as a solvent according to the percolation method under the section on fluid extracts and extracts (Appendix I0). 950ml of percolate is collected. Dragon's blood and borneol are dissolved in 50ml of ethanol, combined with the above percolate, mixed evenly, allowed to stand for 48 hours, filtered, and 1000ml is prepared.

[0004] The traditional preparation method described above only uses simple percolation extraction, which not only takes a long time to extract, but also has a limited dissolution rate of active ingredients and a narrow range of active ingredients, resulting in limited product efficacy, slow onset of action, and a long treatment cycle. Summary of the Invention

[0005] In view of the above-mentioned shortcomings in the prior art, the purpose of this invention is to provide a new process for preparing bone pain relief tincture. The bone pain relief tincture prepared by this new process can significantly increase the content of effective ingredients in the final product, thereby significantly improving the therapeutic effect and shortening the treatment cycle.

[0006] To achieve the above objectives, the solution adopted by the present invention is as follows:

[0007] A new process for a bone pain relief tincture includes: (1) ultra-high pressure extraction: adding 45-55% ethanol to Artemisia argyi for ultra-high pressure treatment, filtering to obtain extract and residue; (2) grinding dried ginger, myrrh, and frankincense into coarse powder and mixing with residue, then adding 50% ethanol for percolation, collecting the percolate and combining it with the extract; separately dissolving dragon's blood and borneol in ethanol, combining with the percolate and extract, mixing, standing, filtering, and obtaining the product.

[0008] The beneficial effects of the new process for bone pain relief tincture provided by this invention are:

[0009] (1) The new process provided by the present invention uses enzymatic hydrolysis under specific conditions to degrade certain substances in the cell wall and intercellular matrix of Artemisia argyi, thereby reducing the resistance of the mass transfer barrier such as cell wall and intercellular matrix when alkaloids diffuse from the cell to the extraction medium, opening the channel for subsequent alkaloid extraction. The reaction conditions are mild, the extraction time is short, and the extraction rate is high.

[0010] (2) The new process provided by the present invention uses enzymatic hydrolysis under specific conditions followed by ultra-high pressure extraction under specific conditions, which can significantly improve the dissolution of effective components in Saussurea costatum. Compared with enzymatic hydrolysis or ultra-high pressure extraction alone, it has a very outstanding extraction effect.

[0011] (3) The new process provided by this invention allows for ultra-high pressure extraction at room temperature without damaging the activity of the extracted active ingredient alkaloids, ensuring that the alkaloids are significantly dissolved while protecting their activity.

[0012] (4) The new process provided by the present invention adopts a step-by-step concentration of ethanol to extract the effective components of Saussurea costatum in the ultra-high pressure stage, which can not only improve the dissolution range of the effective components, but also shorten the subsequent percolation time and improve production efficiency. Detailed Implementation

[0013] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0014] The following is a detailed description of a new process for a bone pain relief tincture provided by an embodiment of the present invention.

[0015] A new process for a bone pain relief tincture includes:

[0016] (1) Bio-enzymatic hydrolysis: Grind *Saussurea involucrata* into powder with a particle size of 0.5-1.0 cm. Add 1-2 times the weight of deionized water to the *Saussurea involucrata* powder to obtain a mixed slurry. Then add 0.5-0.8% of the mixed slurry to a compound enzyme with a concentration of 0.8-1.1 g / L (by weight, the compound enzyme includes 1-2 parts of cellulase and 0.5-0.8 parts of pectinase). Adjust the hydrolysis temperature to 48-52℃ and the pH value to 3.5-4.5. Hydrolyze for 2-3 hours to obtain a mixed solution. Then heat the solution to 90℃ for 4-5 minutes to inactivate the enzyme.

[0017] Under the specific enzymatic hydrolysis conditions described above, the degradation rate of certain substances in the cell wall and intercellular matrix of *Saussurea involucrata* can be significantly increased, which is beneficial to significantly improving the extraction rate of alkaloids from *Saussurea involucrata*.

[0018] (2) Ultra-high pressure extraction: According to the ratio of first ethanol to Saussurea involucrata 2-3ml:1g, add first ethanol with a concentration of 45-50% to the mixture and put it into an ultra-high pressure container. Perform the first ultra-high pressure treatment for 2-4min at room temperature with an ultra-high pressure of 250-350MPa, and filter to obtain the first filtrate and the first filter residue.

[0019] According to the ratio of second ethanol to first filter residue of 2-3 ml: 1 g, add 50-55% second ethanol to the first filter residue and put it into an ultra-high pressure container. Perform a second ultra-high pressure treatment at 350-430 MPa at room temperature for 2-4 min, and filter to obtain the second filtrate and drug residue.

[0020] Combine the first and second filtrates to obtain the extract.

[0021] In this embodiment, under the specific ultra-high pressure extraction conditions described above, ultra-high pressure extraction of enzymatically hydrolyzed Artemisia capillaris with ethanol at stepwise concentrations can facilitate the full dissolution of the effective components in Artemisia capillaris.

[0022] (3) Percolation: Grind dried ginger, myrrh, and frankincense into coarse powder and mix them with the dregs. Then, continue percolation with 50% ethanol at a flow rate of 8-10 ml / min. Collect the percolate and combine it with the extract to obtain 950 ml. Separately, dissolve dragon's blood and borneol in 50 ml of ethanol, combine it with the percolate and extract, mix well, let stand for 48 h, filter, and make 1000 ml.

[0023] In this embodiment, the effective substances in Saussurea costatum are fully dissolved through a combination of enzymatic extraction and ultra-high pressure extraction under the specific conditions described above. In the percolation step, the percolation flow rate can be increased, thereby shortening the production time and reducing time costs.

[0024] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0025] Example 1

[0026] This embodiment provides a new process for bone pain relief tincture, including:

[0027] (1) Bio-enzymatic hydrolysis: Grind Artemisia argyi into powder to a particle size of 0.8 cm. Add 1.5 times the weight of deionized water to the Artemisia argyi powder to obtain a mixed slurry. Then add 0.7% of the mixed slurry to a compound enzyme with a concentration of 1 g / L (by weight, the compound enzyme includes 1.5 parts of cellulase and 0.6 parts of pectinase). Adjust the hydrolysis temperature to 50℃ and the pH value to 3.8. Hydrolyze for 1.5 h to obtain a mixed solution. Then heat to 90℃ for inactivation treatment for 4.5 min.

[0028] (2) Ultra-high pressure extraction: According to the ratio of first ethanol to Saussurea involucrata as 2.5ml:1g, first ethanol with a concentration of 48% was added to the mixture and placed in an ultra-high pressure container. The first ultra-high pressure treatment was carried out at 320MPa at room temperature for 3min. The first filtrate and the first filter residue were obtained by filtration.

[0029] The ratio of second ethanol to first filter residue was 2.5 ml: 1 g. 52% second ethanol was added to the first filter residue and placed in an ultra-high pressure container. The container was subjected to a second ultra-high pressure treatment at 380 MPa at room temperature for 3 min. The second filtrate and the residue were obtained by filtration. The first filtrate and the second filtrate were combined to obtain the extract.

[0030] (3) Percolation: Grind dried ginger, myrrh, and frankincense into coarse powder and mix them with the dregs. First, percolate with the extract, and then continue percolation with 50% ethanol at a flow rate of 9 ml / min. Collect the percolate and combine it with the extract to obtain 950 ml. Separately, dissolve dragon's blood and borneol in 50 ml of ethanol, combine it with the percolate and extract, mix well, let stand for 48 h, filter, and make 1000 ml.

[0031] Example 2

[0032] This embodiment provides a new process for bone pain relief tincture, including: (1) biological enzymatic hydrolysis: grinding Artemisia argyi powder to a particle size of 0.5 cm, adding 1 times the weight of deionized water to Artemisia argyi powder to obtain a mixed slurry, then adding 0.5% of the mass of a compound enzyme with a concentration of 1.1 g / L (by weight, the compound enzyme includes: 1.5 parts cellulase and 0.6 parts pectinase), adjusting the enzymatic hydrolysis temperature to 48°C, the pH value to 3.5, enzymatic hydrolysis for 2 h, obtaining a mixed solution, and then heating to 90°C for 5 min for inactivation treatment.

[0033] The rest is the same as in Example 1.

[0034] Example 3

[0035] This embodiment provides a new process for bone pain relief tincture, including: (1) biological enzymatic hydrolysis: grinding Artemisia argyi powder to a particle size of 1.0 cm, adding twice the weight of deionized water to Artemisia argyi powder to obtain a mixed slurry, then adding 0.8% of the mass of a compound enzyme with a concentration of 0.8 g / L (by weight, the compound enzyme includes: 1.5 parts cellulase and 0.6 parts pectinase), adjusting the enzymatic hydrolysis temperature to 52℃, the pH value to 4.5, enzymatic hydrolysis for 1 h, obtaining a mixed solution, and then heating to 90℃ for 4 min for inactivation treatment.

[0036] The rest is the same as in Example 1.

[0037] Example 4

[0038] This embodiment provides a new process for bone pain relief tincture, including: improving step (2), the rest being the same as in embodiment 1;

[0039] (2) Ultra-high pressure extraction: According to the ratio of first ethanol to Saussurea involucrata 2ml:1g, first ethanol with a concentration of 45% was added to the mixture and placed in an ultra-high pressure container. The first ultra-high pressure treatment was carried out at 350MPa at room temperature for 2min. The first filtrate and the first filter residue were obtained by filtration.

[0040] According to the ratio of second ethanol to first filter residue of 3ml:1g, 50% second ethanol was added to the first filter residue and placed in an ultra-high pressure container. The container was subjected to a second ultra-high pressure treatment at 430MPa at room temperature for 2min. The second filtrate and the drug residue were obtained by filtration. The first filtrate and the second filtrate were combined to obtain the extract. The drug residue was dried.

[0041] Example 5

[0042] This embodiment provides a new process for bone pain relief tincture, including: improving step (2), the rest being the same as in embodiment 1;

[0043] (2) Ultra-high pressure extraction: According to the ratio of first ethanol to Saussurea involucrata 3ml:1g, add 50% first ethanol to the mixture and put it into an ultra-high pressure container. Perform the first ultra-high pressure treatment for 4min at room temperature with an ultra-high pressure of 250MPa. Filter to obtain the first filtrate and the first filter residue.

[0044] The ratio of second ethanol to first filter residue was 2 ml: 1 g. 55% second ethanol was added to the first filter residue and placed in an ultra-high pressure container. The container was subjected to a second ultra-high pressure treatment at 350 MPa at room temperature for 4 min. The second filtrate and the residue were obtained by filtration. The first filtrate and the second filtrate were combined to obtain the extract.

[0045] Example 6

[0046] This embodiment provides a new process for bone pain relief tincture, including: (1) biological enzymatic hydrolysis: by weight, the compound enzyme includes: 1 part cellulase and 0.8 parts pectinase.

[0047] The rest is the same as in Example 1.

[0048] Example 7

[0049] This embodiment provides a new process for bone pain relief tincture, including: (1) biological enzymatic hydrolysis: by weight, the compound enzyme includes: 2 parts cellulase and 0.5 parts pectinase.

[0050] The rest is the same as in Example 1.

[0051] Example 8

[0052] This embodiment provides a new process for bone pain relief tincture, including: improving step (2), the rest being the same as in embodiment 1;

[0053] (2) Ultra-high pressure extraction: According to the ratio of ethanol to Saussurea costatum of 7.5ml:1g, add 50% ethanol to Saussurea costatum and place it in an ultra-high pressure container. Under high pressure of 400MPa at room temperature for 9min, filter to obtain extract and residue.

[0054] Example 9

[0055] This embodiment provides a new process for bone pain relief tincture, including:

[0056] (1) Ultra-high pressure extraction: Grind the snow elm powder to a particle size of 0.8 cm, add 48% first ethanol to the mixture according to the ratio of first ethanol to snow elm of 2.5 ml: 1 g, put it into an ultra-high pressure container, and carry out the first ultra-high pressure treatment for 3 min at room temperature with an ultra-high pressure of 320 MPa. Filter to obtain the first filtrate and the first filter residue.

[0057] The ratio of second ethanol to first filter residue was 2.5 ml: 1 g. 52% second ethanol was added to the first filter residue and placed in an ultra-high pressure vessel. The vessel was subjected to a second ultra-high pressure treatment at 380 MPa at room temperature for 3 min. The second filtrate and the residue were obtained by filtration. The first filtrate and the second filtrate were combined to obtain the extract.

[0058] (2) Percolation: Grind dried ginger, myrrh, and frankincense into coarse powder and mix them with the dregs. Then, continue percolation with 50% ethanol at a flow rate of 9 ml / min. Collect the percolate and combine it with the extract to obtain 950 ml. Separately, dissolve dragon's blood and borneol in 50 ml of ethanol, combine it with the percolate and extract, mix well, let stand for 48 h, filter, and make 1000 ml.

[0059] Comparative Example 1

[0060] This comparative example provides a new process for bone pain relief tincture, including:

[0061] (1) Bio-enzymatic hydrolysis: Grind Artemisia argyi into powder to a particle size of 0.8 cm. Add 1.5 times the weight of deionized water to the Artemisia argyi powder to obtain a mixed slurry. Then add 0.7% of the mixed slurry to a compound enzyme with a concentration of 1 g / L (by weight, the compound enzyme includes 1.5 parts cellulase and 0.6 parts pectinase). Adjust the enzymatic hydrolysis temperature to 50℃ and the pH value to 3.8. After enzymatic hydrolysis for 1.5 h, the mixture is heated to 90℃ for 4.5 min for inactivation treatment. The residue is then filtered to obtain the medicinal residue.

[0062] (2) Percolation: Grind dried ginger, myrrh, and frankincense into coarse powder and mix them with the dregs. Percolate with 50% ethanol at a flow rate of 9 ml / min and collect 950 ml of percolate. Separately, dissolve dragon's blood and borneol in 50 ml of ethanol, combine them with the percolate, mix well, let stand for 48 h, filter, and make 1000 ml of the solution.

[0063] Comparative Example 2

[0064] This comparative example provides a new process for bone pain relief tincture, including: (1) biological enzymatic hydrolysis: the enzyme used for enzymatic hydrolysis is cellulase.

[0065] The rest is the same as in Example 1.

[0066] Comparative Example 3

[0067] This comparative example provides a new process for bone pain relief tincture, including: (1) biological enzymatic hydrolysis: the enzyme used for enzymatic hydrolysis is pectinase.

[0068] The rest is the same as in Example 1.

[0069] Comparative Example 4

[0070] This comparative example provides a new process for bone pain relief tincture, including: (1) biological enzymatic hydrolysis: grinding Artemisia argyi powder to a particle size of 0.8 cm, adding 3 times the weight of deionized water to Artemisia argyi powder to obtain a mixed slurry, then adding 1% of the mass of a 1.5 g / L complex enzyme (by weight, the complex enzyme includes: 1.5 parts cellulase and 0.6 parts pectinase), adjusting the enzymatic hydrolysis temperature to 45℃, the pH value to 4.8, enzymatic hydrolysis for 2.5 h, obtaining a mixed solution, and then heating to 90℃ for inactivation treatment for 4.5 min.

[0071] The rest is the same as in Example 1.

[0072] Comparative Example 5

[0073] This comparative example provides a new process for bone pain relief tincture, including: improving step (2), the rest being the same as in Example 1;

[0074] (2) Ultra-high pressure extraction: According to the ratio of first ethanol to Saussurea involucrata as 1.5ml:1g, first ethanol with a concentration of 52% was added to the mixture and placed in an ultra-high pressure container. The first ultra-high pressure treatment was carried out at 400MPa at room temperature for 3min. The first filtrate and the first filter residue were obtained by filtration.

[0075] The ratio of second ethanol to first filter residue was 3.5 ml: 1 g. 58% second ethanol was added to the first filter residue and placed in an ultra-high pressure container. The container was subjected to a second ultra-high pressure treatment at 450 MPa at room temperature for 3 min. The second filtrate and the residue were obtained by filtration. The first filtrate and the second filtrate were combined to obtain the extract.

[0076] Comparative Example 6

[0077] This comparative example provides a new process for bone pain relief tincture, including: (1) biological enzymatic hydrolysis: by weight, the compound enzyme includes: 0.8 parts cellulase and 1 part pectinase.

[0078] The rest is the same as in Example 1.

[0079] Experimental Example 1

[0080] Experimental methods:

[0081] 180 rats were randomly divided into a normal group, a model group, a bone pain relief tincture group, and drug administration groups 1-15 (corresponding to Examples 1-9 and Comparative Examples 1-6), with 10 rats per group.

[0082] The drugs were administered twice, once 1 day before modeling and once immediately after modeling. The right hind limb toes of rats in the normal and model groups were immersed in 10 mL of physiological saline, once every 5 minutes, for a total of 6 immersions, each lasting 5 minutes. The right hind limb toes of rats in the bone pain relief tincture group were immersed in 10 mL of bone pain relief tincture, once every 5 minutes, for a total of 6 immersions. The right hind limb toes of rats in treatment groups 1-15 were immersed in 10 mL of the combined drug solution provided in Examples 1-9 and Comparative Examples 1-6, respectively, once every 5 minutes, for a total of 6 immersions.

[0083] One day after drug administration, the normal group was injected with an equal volume of physiological saline, while the other groups received a subcutaneous injection of 1% λ-carrageenan saline suspension into the mid-plantar portion of the right hind limb of each rat, with an injection volume of 0.1 mL per rat. Immediately after model establishment, each group underwent another administration of the drug as described above. At 0.5, 1, and 2 hours after model establishment, the paw volume of each group was measured using a paw volume analyzer.

[0084] Toe swelling (mL) = Post-injection toe volume - baseline volume;

[0085] Toe swelling inhibition rate (%) = (swelling degree of model group - swelling degree of drug group) / swelling degree of model group × 100%.

[0086] Five hours after modeling, the rats were anesthetized with sodium pentobarbital and sacrificed. Subcutaneous tissue was collected from the dorsal side of the right hind paw of the rats, temporarily stored in liquid nitrogen, and then transferred to a -80°C freezer for preservation. After tissue homogenization, TNF-α (pg·mL⁻¹) was detected using a rat ELISA kit. -1 ), 1L-1β (pg·mL) -1 The levels of the test were shown in Table 1.

[0087] Table 1

[0088]

[0089] As shown in Table 1, the Bone Pain Relief Tincture had a significant effect 1 hour after modeling and administration, while the treatment groups 1-9 significantly reduced the swelling of rat paws within 0.5 hours after modeling and administration (p<0.05). This indicates that the Bone Pain Relief Tincture prepared using the new process provided in Examples 1-9 has a significantly improved anti-swelling effect compared to the Bone Pain Relief Tincture prepared by existing technologies.

[0090] Compared with the normal group, the levels of 1L-1β and TNF-α in the model group were significantly increased, indicating that the modeling induced a significant inflammatory response. Compared with the model group, the drug administration groups 1-9 showed a significant inhibitory effect on the release of TNF-α and 1L-1β (p<0.05), and the inhibitory effect was significantly better than that of the bone pain relief tincture group. This indicates that the bone pain relief tincture prepared using the new process provided in Examples 1-9 has a significantly improved anti-inflammatory effect compared with the bone pain relief tincture prepared by existing technologies.

[0091] Example 2

[0092] Experimental methods: 170 mice were randomly divided into model group, bone pain relief tincture group and drug administration group 1-15 (corresponding to Examples 1-9 and Comparative Examples 1-6), with 10 mice per group.

[0093] Pour 1 ml of the provided medicine solution from each group (excluding the model group) onto a 2 cm layer of non-woven fabric. 2 (1*2cm) After hair removal with 8% sodium sulfide on the abdomen of mice, non-woven fabric was applied to each mouse, covered with two layers of sulfuric acid paper, and sealed and fixed with non-irritating adhesive tape. Each group was treated once a day for 5 hours each time. One hour after the last administration, each group of mice was injected intraperitoneally with 0.6% acetic acid (0.1ml / 10g) solution (0.2ml / mouse) to establish the model.

[0094] The number of times the mice twisted their bodies within 15 minutes was recorded (based on abdominal concavity, hind limb extension, and hip elevation). The results are shown in Table 2.

[0095] Table 2

[0096]

[0097] Table 2 shows that, compared with the model group, the treatment groups 1-9 significantly reduced the number of acetic acid-induced writhing movements in mice (p<0.05), and had a very significant analgesic effect compared with the bone pain relief tincture group. This indicates that the bone pain relief tincture prepared using the new process provided in Examples 1-9 has a significantly improved analgesic effect compared with the bone pain relief tincture prepared by existing technologies.

[0098] In summary, the new process for preparing Gutongling Tincture provided by this invention can significantly increase the content of effective ingredients in the final product, thereby significantly improving the therapeutic effect and shortening the treatment cycle.

[0099] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A new process for a bone pain relief tincture, characterized in that: include: (1) Bio-enzymatic hydrolysis and ultra-high pressure extraction: The *Saussurea involucrata* powder is obtained by grinding the *Saussurea involucrata* powder, and a mixed solution is obtained by enzymatic hydrolysis using a compound enzyme, which consists of 1-2 parts cellulase and 0.5-0.8 parts pectinase. The enzymatic hydrolysis process is as follows: 1-2 times the weight of deionized water is added to the *Saussurea involucrata* powder to obtain a mixed slurry, and then 0.5-0.8% of the compound enzyme is added to the mixed slurry. The enzymatic hydrolysis temperature is adjusted to 48-52℃, the pH value is 3.5-4.5, and the enzymatic hydrolysis is carried out for 2-3 hours to obtain the mixed solution. In the ultra-high pressure extraction, a 45-50% concentration of first ethanol is added to the mixture and placed in an ultra-high pressure vessel for a first ultra-high pressure treatment. The mixture is then filtered to obtain a first filtrate and a first filter residue. A 50-55% concentration of second ethanol is added to the first filter residue and placed in an ultra-high pressure vessel for a second ultra-high pressure treatment. The mixture is then filtered to obtain a second filtrate and a residue. The first and second filtrates are combined to obtain the extract. The ratio of the first ethanol to the *Saussurea involucrata* is 2-3 ml:1 g. The conditions for the first ultra-high pressure treatment are: ultra-high pressure of 250-350 MPa, and treatment at room temperature for 2-4 minutes. The ratio of the second ethanol to the first filter residue is also 2-3 ml:1 g. The conditions for the second ultra-high pressure treatment are: ultra-high pressure of 350-430 MPa, and treatment at room temperature for 2-4 minutes. (2) Percolation: Grind dried ginger, myrrh, and frankincense into coarse powder and mix them with the dregs. Then add 50% ethyl acetate. The alcohol is used for percolation, the percolate is collected and combined with the extract; separately, dragon's blood and borneol are dissolved in the ethanol, combined with the percolate and the extract, mixed well, allowed to stand, and filtered to obtain the final product.

2. The new process for bone pain relief tincture according to claim 1, characterized in that: The concentration of the complex enzyme is 0.8- 1.1g / l。 3. The new process for bone pain relief tincture according to claim 1, characterized in that: After obtaining the mixture, the temperature is raised to... Inactivate at 90℃ for 4-5 minutes.

4. The new process for bone pain relief tincture according to claim 1, characterized in that: The powder of *Artemisia annua* The diameter is 0.5-1.0cm.

Images