Traditional Chinese medicine composition for treating chronic pelvic inflammation as well as preparation method and detection method thereof

Abstract

The invention belongs to the technical field of pharmaceutical compositions, and particularly provides a traditional Chinese medicine composition for treating chronic pelvic inflammation as well as a preparation method and a detection method thereof. The traditional Chinese medicine composition comprises 400-450 g of folium artemisiae argyi, 10-50 g of angelica sinensis, 300-400 g of sargentgloryvine stem, 10-50 g of wine-processed teasel root, 10-50 g of obscured homalomena rhizome, 10-50 g of vinegar-processed myrrh, 10-50 g of radix angelicae pubescentis, 10-50 g of radix paeoniae rubra, 10-50 g of dragon's blood, 10-50 g of cortex periplocae, 10-50 g of radix angelicae, 10-50 g of radix saposhnikoviae, 10-50 g of safflower carthamus, 10-50 g of mistletoe, 10-50 g of fried pepper, 10-50 g of vinegar-processed frankincense and 10- The traditional Chinese medicine has the characteristics of reinforcing qi, nourishing blood, nourishing liver and tonifying kidney; the traditional Chinese medicine composition has the effects of promoting qi and blood circulation, tonifying liver and kidney, strengthening vital qi, eliminating the pathogenic factors and recovering the pathogenic factors.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical composition technology, and relates to a traditional Chinese medicine composition for treating chronic pelvic inflammatory disease, its preparation method, and its detection method. Background Technology

[0002] Chronic pelvic inflammatory disease is a common and frequently occurring gynecological disease. In traditional Chinese medicine, it falls under the categories of "abdominal pain in women," "abdominal masses," and "leukorrhea." It refers to inflammation of the female internal reproductive organs and surrounding connective tissue and pelvic peritoneum. It is often caused by acute pelvic inflammatory disease that has not been treated in time or thoroughly, leading to long-term congestion and edema of pelvic tissues, thickening of inflamed tissues, and local blood circulation disorders.

[0003] Chronic pelvic inflammatory disease is usually accompanied by long-term chronic pelvic pain and may even lead to serious complications such as infertility and ectopic pregnancy. It mainly occurs in women of childbearing age. Relevant epidemiological statistics show that the incidence rate in women of childbearing age is as high as 10% to 15%, seriously endangering the life, health and safety and quality of life of female patients.

[0004] Traditional Chinese medicine believes that the pelvic cavity is located in the lower abdomen, where the uterus and the Chong, Ren, and Dai meridians, which obtain essential nutrients and perform their functions, all circulate and converge. When pathogenic factors invade through the vulva and obstruct the uterus and its meridians, the flow of Qi and blood in the uterus is inevitably hindered, leading to stagnation and ultimately "blood stasis." The *Jinkui Yaolue* states, "When blood is not flowing smoothly, it becomes water," indicating that blood stasis affects the normal vaporization of fluids, causing fluid to accumulate in the lower abdomen, resulting in leukorrhea. Furthermore, water accumulation disrupts the Qi mechanism, leading to Qi stagnation, impaired blood flow, and ultimately, blood stasis. "Where there is no flow, there is pain," manifesting primarily as abdominal pain. Treatment focuses on strengthening the body's resistance, supplemented by eliminating pathogenic factors. Summary of the Invention

[0005] The present invention aims to provide a traditional Chinese medicine composition, preparation method and detection method for treating chronic pelvic inflammatory disease of the qi stagnation and blood stasis and damp-heat accumulation type.

[0006] This invention provides a traditional Chinese medicine composition for treating chronic pelvic inflammatory disease, comprising: 400-450g of Artemisia argyi, 10-50g of Angelica sinensis, 300-400g of Sargentodoxa cuneata, 10-50g of Dipsacus asper (processed with wine), 10-50g of Homalomena occulta, 10-50g of Commiphora myrrha (processed with vinegar), 10-50g of Angelica pubescens, 10-50g of Paeonia lactiflora, 10-50g of Daemonorops draco, 10-50g of Periploca sepium, 10-50g of Angelica dahurica, 10-50g of Saposhnikovia divaricata, 10-50g of Carthamus tinctorius, 10-50g of Loranthus parasiticus, 10-50g of Zanthoxylum bungeanum (fried), 10-50g of Boswellia carterii (processed with vinegar), and 10-50g of Notopterygium incisum. in: Artemisia argyi and Sargentodoxa cuneata are the principal herbs; The main ingredients are safflower, frankincense, myrrh, red peony root, angelica root, and dragon's blood. The herbs used as adjuvants are Angelica pubescens, Notopterygium incisum, Loranthus parasiticus, Dipsacus asper, Periploca sepium, Homalomena occulta, Saposhnikovia divaricata, Angelica dahurica, and Zanthoxylum bungeanum.

[0007] Furthermore, the traditional Chinese medicine composition for treating chronic pelvic inflammatory disease includes 427g of Artemisia argyi, 20g of Angelica sinensis, 343g of Sargentodoxa cuneata, 20g of Dipsacus asper (processed with wine), 10g of Homalomena occulta, 20g of Myrrh (processed with vinegar), 10g of Angelica pubescens, 20g of Paeonia lactiflora, 10g of Daemonorops draco, 20g of Periploca sepium, 10g of Angelica dahurica, 20g of Saposhnikovia divaricata, 10g of Carthamus tinctorius, 20g of Loranthus parasiticus, 10g of Zanthoxylum bungeanum (fried), 20g of Boswellia carterii (processed with vinegar), and 10g of Notopterygium incisum.

[0008] Furthermore, the dosage form of the traditional Chinese medicine composition is: granules, capsules, or powder.

[0009] As a further aspect of the present invention, the present invention also provides a method for preparing a traditional Chinese medicine composition for treating chronic pelvic inflammatory disease, comprising the following steps: Step 1: Take a portion of mugwort leaves and put them into a grading ultrafine continuous water-cooled pulverizer with a 100-mesh sieve for pulverization. After passing through a second sieve, you will get coarse mugwort powder. Take one part each of Angelica sinensis, Sargentodoxa cuneata, Dipsacus asper, Homalomena occulta, Myrrh, Angelica pubescens, Paeonia lactiflora, Daemonorops draco, Periploca sepium, Angelica dahurica, Saposhnikovia divaricata, Carthamus tinctorius, Mistletoe, Sichuan pepper, Boswellia carterii, and Notopterygium incisum, and put them into a grading ultrafine continuous water-cooled pulverizer with a 100-mesh sieve for pulverization. After passing through a No. 6 sieve, a mixed fine powder is obtained. Step 2: Mix the coarse powder of Artemisia argyi and the fine powder, and pass them through a No. 2 sieve to obtain a powder for treating chronic pelvic inflammatory disease.

[0010] Furthermore, the specific process for obtaining coarse mugwort powder is as follows: Take a portion of mugwort leaves and place them in a 100-mesh sieve for the first pulverization in a continuous water-cooled ultrafine pulverizer to obtain the first pulverized powder. The first-crushed powder is placed in a 100-mesh sieve and subjected to a second-crushing process in a continuous water-cooled ultrafine pulverizer to obtain the second-crushed powder. The powder was pulverized a second time and passed through a No. 2 sieve to obtain coarse Artemisia argyi powder.

[0011] As a further aspect of the present invention, the present invention also provides a method for detecting a powder for treating chronic pelvic inflammatory disease as described above, comprising: (i) The content of paeoniflorin was used as an indicator for determining the uniformity of mixing. (ii) Determination of paeoniflorin in Paeonia lactiflora according to the standard in Part I of the 2020 edition of the Chinese Pharmacopoeia; Chromatographic conditions: Column: Kromasil 100-5-C18; Mobile phase A: Acetonitrile-0.1% phosphoric acid aqueous solution; Mobile phase B: acetonitrile, gradient elution; Flow rate: 1.0 ml / min; Detection wavelength: 230nm; Column temperature: 35℃; Injection volume: 10 µl; Extraction time investigation: Take 1g of the powder and place it in a stoppered conical flask, add 20ml of methanol; weigh it and soak for 4 hours; sonicate for 10 minutes, 20 minutes or 30 minutes; cool it and weigh it again; make up the weight loss with methanol, shake well, filter, and take the filtrate to obtain the product; (III) Validation of the paeoniflorin analytical method; Blank solvent: methanol; 0.5 mg / ml reference stock solution: Weigh approximately 10 mg of paeoniflorin reference standard accurately, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with methanol, and shake well to obtain the solution. Negative control: Take about 0.43g of Artemisia argyi powder and about 0.34g of Sargentodoxa cuneata powder, weigh them accurately, put them in the same stoppered conical flask, add 20ml of methanol accurately, weigh them, soak for 4 hours, sonicate for 20 minutes, cool, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the negative control. Test solution: Take about 1g of powder, weigh accurately, place it in a stoppered conical flask, add 20ml of methanol accurately, weigh, soak for 4 hours, sonicate for 20 minutes, cool, weigh again, replenish the lost weight with methanol, shake well, filter, and take the filtrate to obtain the test solution. The solution described in (iii) was determined according to the provisional chromatographic conditions; (iv) Examination of linear relationships; Reference solution 18 μg / ml: Accurately transfer 3.6 ml of the reference stock solution into a 100 ml volumetric flask, dilute to the mark with methanol, and shake well; (v) Precision; Take the reference solution and inject it for analysis under the provisional chromatographic conditions. The injection volume is 2 μl. Repeat the analysis 6 times and record the peak area. (vi) Repetitiveness; Take 6 samples from the same batch, prepare test solutions according to the provisional test solution preparation method, inject and analyze according to the provisional chromatographic conditions, record the peak area, and calculate the paeoniflorin content; (vii) Stability test; Take an appropriate amount of any test solution under the repeatability test, and inject it for analysis at room temperature for 0h, 2h, 6h, 12h, 18h and 24h according to the determined chromatographic conditions, and record the peak area. (viii) Accuracy test; Take 1g of sample, accurately weigh it, and divide it into 6 portions. Add 1ml of reference stock solution to each portion accurately. Prepare the sample according to the provisional test solution preparation method, inject and determine the sample under the provisional chromatographic conditions, and calculate the recovery rate.

[0012] Furthermore, the ratio between acetonitrile and 0.1% phosphoric acid aqueous solution was set to 20:80.

[0013] Furthermore, the specific process of gradient elution is as follows: 0~10min100%A, 10~11min100%A-10%A, 11~21min10%A, 21~30min100%A.

[0014] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a traditional Chinese medicine composition for treating chronic pelvic inflammatory disease, which has the characteristics of invigorating qi and nourishing blood, and tonifying the liver and kidneys. The composition uses Artemisia argyi and Sargentodoxa cuneata as the principal herbs, Carthamus tinctorius, Boswellia carterii, Commiphora myrrha, Paeonia lactiflora, Angelica sinensis, and Dracaena cochinchinensis as the assistant herbs, and Angelica pubescens, Notopterygium incisum, Loranthus parasiticus, Dipsacus asper, Periploca sepium, Homalomena occulta, Saposhnikovia divaricata, Angelica dahurica, and Zanthoxylum bungeanum as adjuvant herbs. This composition eliminates stagnation and dampness, promotes the flow of qi and blood, tonifies the liver and kidneys, replenishes the body's vital energy, and restores the body's balance after the elimination

[0015] In addition to the objectives, features, and advantages described above, the present invention has other objectives, features, and advantages. The invention will now be described in further detail with reference to the figures. Attached Figure Description

[0016] The accompanying drawings, which form part of this application, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings: Figure 1 This is a chromatogram under blank solvent conditions in the first test of the analysis method of paeoniflorin in powder in Example 3 of the present invention; Figure 2 This is a chromatogram under the negative control conditions of the first test of the analysis method of paeoniflorin in powder in Example 3 of the present invention; Figure 3 This is a chromatogram of paeoniflorin stock solution under the conditions of the first investigation of the paeoniflorin analysis method in powder in Example 3 of the present invention; Figure 4 This is the chromatogram of the test sample solution under the test sample solution conditions in the first test of the analysis method of paeoniflorin in powder in Example 3 of the present invention; Figure 5 This is the chromatogram under blank solvent conditions for the second test of the analysis method of paeoniflorin in powder in Example 3 of the present invention; Figure 6 This is a chromatogram under the negative control conditions of the second investigation of the analysis method of paeoniflorin in powder in Example 3 of the present invention; Figure 7 This is a chromatogram of paeoniflorin stock solution under the conditions of the second investigation of the paeoniflorin analysis method in powder in Example 3 of the present invention; Figure 8 This is the chromatogram of the test sample solution under the test sample solution conditions in the second investigation of the analysis method of paeoniflorin in powder in Example 3 of the present invention; Figure 9 This is the linear regression diagram of paeoniflorin in Example 3 of the present invention. Detailed Implementation

[0017] To make the above-mentioned objectives, features, and advantages of the present invention clearer and easier to understand, the specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. It should be noted that the accompanying drawings of the present invention are all in a simplified form and use non-precise proportions, and are only used to facilitate and clearly assist in illustrating the implementation of the present invention; the "several" mentioned in the present invention are not limited to the specific number shown in the examples in the accompanying drawings; the orientations or positional relationships indicated by terms such as "front," "middle," "rear," "left," "right," "up," "down," "top," "bottom," and "center" mentioned in the present invention are all based on the orientations or positional relationships shown in the accompanying drawings of the present invention, and do not indicate or imply that the device or component referred to must have a specific orientation, nor should they be construed as limitations on the present invention.

[0018] Example 1: The present invention provides a traditional Chinese medicine composition for treating chronic pelvic inflammatory disease, comprising: 400-450g of Artemisia argyi, 10-50g of Angelica sinensis, 300-400g of Sargentodoxa cuneata, 10-50g of Dipsacus asper (processed with wine), 10-50g of Homalomena occulta, 10-50g of Commiphora myrrha (processed with vinegar), 10-50g of Angelica pubescens, 10-50g of Paeonia lactiflora, 10-50g of Daemonorops draco, 10-50g of Periploca sepium, 10-50g of Angelica dahurica, 10-50g of Saposhnikovia divaricata, 10-50g of Carthamus tinctorius, 10-50g of Loranthus parasiticus, 10-50g of Zanthoxylum bungeanum (fried), 10-50g of Boswellia carterii (processed with vinegar), and 10-50g of Notopterygium incisum.

[0019] Artemisia argyi and Sargentodoxa cuneata are the principal herbs; among which: Artemisia argyi can warm the blood and meridians, making it an essential medicine for treating coldness in the uterus in gynecology. Modern pharmacological studies have shown that it has analgesic, anti-inflammatory, and hemostatic effects. The herb *Sargentodoxa cuneata* clears heat, invigorates blood circulation, and relieves pain. Modern pharmacological studies have shown that it has inhibitory effects on *Escherichia coli*, *Pseudomonas aeruginosa*, and other bacteria, and can also inhibit platelet aggregation.

[0020] The herbs used as assistant herbs are safflower, frankincense, myrrh, red peony root, angelica root, and dragon's blood; among them: Angelica sinensis is pungent and warm, making it an excellent medicine for promoting blood circulation and removing blood stasis, as well as a holy medicine for nourishing blood. Modern pharmacological studies have shown that it has the effects of inhibiting uterine contractions, anti-inflammatory and analgesic effects. Safflower promotes blood circulation, regulates menstruation, disperses blood stasis, and relieves pain. It can be used for diseases such as abdominal pain due to blood stasis and abdominal masses. Modern pharmacological studies have shown that it has bidirectional uterine regulation, anti-inflammatory, and analgesic effects. Frankincense and myrrh promote blood circulation, relieve pain and reduce swelling. When used together, they can be used for various pain syndromes caused by blood stasis. Modern pharmacological studies have shown that both have anti-inflammatory, antibacterial and analgesic effects, and myrrh can inhibit uterine smooth muscle contraction. Dragon's blood promotes blood circulation, removes blood stasis, and relieves pain. It also has the effects of dispersing blood stasis and stopping bleeding. It has pharmacological effects of inhibiting platelet aggregation, preventing thrombosis, and anti-inflammatory and analgesic properties. Red peony root clears heat, disperses blood stasis, and relieves pain. It has antipyretic, analgesic, immune-regulating, and antioxidant pharmacological effects.

[0021] The adjuvant herbs are Angelica pubescens, Notopterygium incisum, Loranthus parasiticus, Dipsacus asper, Periploca sepium, Homalomena occulta, Saposhnikovia divaricata, Angelica dahurica, and Zanthoxylum bungeanum; among which: Notopterygium root and Angelica pubescens root dispel wind, dampness, and relieve pain. Both have anti-inflammatory and analgesic effects. Notopterygium root can also relieve fever and has antibacterial properties, while Angelica pubescens root can calm the nerves and inhibit platelet aggregation. The pungent and bitter properties of the herb *Cinnamomum camphora* can dispel wind, its bitter properties can lead to premature death, and its warm properties can dispel cold. It also has a tonic effect. Modern pharmacological studies have shown that it has anti-inflammatory, anti-stress, analgesic, and sedative effects. Both mistletoe and teaspoon have the effects of tonifying the liver and kidneys, strengthening tendons and bones, and relieving pain. Mistletoe has antibacterial, anti-inflammatory, antiviral, and analgesic pharmacological effects. Dipsacus has pharmacological effects such as anti-inflammatory, anti-aging, and antioxidant properties; Qiannianjian dispels rheumatism and strengthens muscles and bones; its clinical efficacy is even better when combined with other herbs such as Angelica pubescens, Loranthus parasiticus, and Periploca sepium. It has the properties of dispelling wind, eliminating dampness, and relieving pain. Modern pharmacology shows that it has antipyretic, anti-inflammatory, analgesic, and anti-allergic effects. Angelica dahurica has the functions of drying dampness and stopping leukorrhea, dispelling cold and wind and relieving pain, and has pharmacological effects such as antipyretic, antifungal and anti-inflammatory effects. Sichuan pepper has warming and pain-relieving properties, and can also inhibit bacteria, relieve pain, and reduce inflammation.

[0022] When used together, these herbs can eliminate blood stasis and dampness, promote the flow of qi and blood, replenish the liver and kidneys, and strengthen the body's vital energy, thus restoring health after the elimination of pathogens.

[0023] Preferably, the traditional Chinese medicine composition is specifically configured as follows: The formula includes 427g of Artemisia argyi, 20g of Angelica sinensis, 343g of Sargentodoxa cuneata, 20g of Dipsacus asper (processed with wine), 10g of Homalomena occulta, 20g of Myrrh (processed with vinegar), 10g of Angelica pubescens, 20g of Paeonia lactiflora, 10g of Dracaena cochinchinensis, 20g of Periploca sepium, 10g of Angelica dahurica, 20g of Saposhnikovia divaricata, 10g of Carthamus tinctorius, 20g of Mistletoe, 10g of Sichuan pepper (fried), 20g of Boswellia carterii (processed with vinegar), and 10g of Notopterygium incisum.

[0024] Preferably, the dosage form of the traditional Chinese medicine composition is granules, capsules, or powder. Specifically, in this embodiment, the dosage form of the traditional Chinese medicine composition is preferably powder.

[0025] Preferably, the safflower, notopterygium root, angelica root, red peony root, licorice root bark, saposhnikovia root, mistletoe, and vinegar-processed frankincense in the traditional Chinese medicine composition are all sourced from medicinal slices purchased from Hengdong County Traditional Chinese Medicine Slices Factory; The ingredients in the Chinese herbal composition, including Artemisia argyi, Sargentodoxa cuneata, Homalomena occulta, Angelica pubescens, Daemonorops draco, Angelica dahurica, stir-fried Sichuan pepper, Dipsacus asper, and vinegar-processed myrrh, are all derived from processed medicinal materials purchased from Hunan Ranruntang Chinese Medicine Co., Ltd. All processed medicinal materials comply with the relevant provisions of the 2020 edition of the Pharmacopoeia of the People's Republic of China, Part I, under the category of each medicinal herb processed material.

[0026] Example 2: This invention also provides a method for preparing a traditional Chinese medicine composition for treating chronic pelvic inflammatory disease, comprising the following steps: Step 1: Take a portion of mugwort leaves and put them into a grading ultrafine continuous water-cooled pulverizer with a 100-mesh sieve for pulverization. After passing through a second sieve, you will get coarse mugwort powder. Take one part each of Angelica sinensis, Sargentodoxa cuneata, Dipsacus asper, Homalomena occulta, Myrrh, Angelica pubescens, Paeonia lactiflora, Daemonorops draco, Periploca sepium, Angelica dahurica, Saposhnikovia divaricata, Carthamus tinctorius, Mistletoe, Sichuan pepper, Boswellia carterii, and Notopterygium incisum, and put them into a grading ultrafine continuous water-cooled pulverizer with a 100-mesh sieve for pulverization. After passing through a No. 6 sieve, a mixed fine powder is obtained. Step 2: Place the coarse powder of Artemisia argyi and the fine powder in a trough mixer and mix for 45 minutes. Then, pass the mixture through a No. 2 sieve and package it to obtain a powder for treating chronic pelvic inflammatory disease.

[0027] The preferred process for obtaining coarse Artemisia argyi powder is as follows: Take a portion of mugwort leaves and place them in a 100-mesh sieve for the first pulverization in a continuous water-cooled ultrafine pulverizer to obtain the first pulverized powder. The first-crushed powder is placed in a 100-mesh sieve and subjected to a second-crushing process in a continuous water-cooled ultrafine pulverizer to obtain the second-crushed powder. The powder was pulverized a second time and passed through a No. 2 sieve to obtain coarse Artemisia argyi powder.

[0028] Example 3: This invention also provides a method for detecting a powder used to treat chronic pelvic inflammatory disease, comprising: (a) Detection of Artemisia argyi powder; Experiment 1: Weigh 100g of mugwort leaves and grind them in a grading ultrafine continuous water-cooled pulverizer (model: XC8605, manufacturer: Wenzhou Dingli Medical Equipment Co., Ltd.) equipped with a 100-mesh sieve. Weigh the powder and pass it through a No. 6 sieve. Weigh the powder that can pass through the sieve. Collect all the powder again, pass it through a No. 2 sieve, and weigh the powder that can pass through the sieve.

[0029] Experiment 2: Weigh 100g of mugwort leaves and spread them evenly on a stainless steel tray. Make two parallel portions and dry them in an oven at 50℃ for 24h and 48h respectively. Then, grind them in a grading ultrafine continuous water-cooled pulverizer equipped with a 100-mesh sieve. Weigh the powder and pass it through a No. 6 sieve. Weigh the powder that can pass through the sieve. Collect all the powder again, pass it through a No. 2 sieve, and weigh the powder that can pass through the sieve.

[0030] Experiment 3: Weigh 100g of mugwort leaves and grind them in a graded ultrafine continuous water-cooled pulverizer equipped with a 100-mesh sieve; collect the powder and grind it again in the graded ultrafine continuous water-cooled pulverizer equipped with a 100-mesh sieve, weigh the powder, pass it through a No. 6 sieve, and weigh the powder that can pass through the sieve; collect all the powder again, pass it through a No. 2 sieve, weigh the powder that can pass through the sieve, pass the powder that can pass through the No. 2 sieve through a No. 4 sieve, and weigh the powder that can pass through the sieve.

[0031] Sieveable powder yield: The sieveable powder yield was calculated. The results are shown in Table 1. ; Note: W—weight of powder after crushing (g); W1—weight of powder that can be sieved (g).

[0032] Table 1: Results of the investigation on the yield of Artemisia argyi powder that can pass through different sieves

[0033] Experimental results show that after being pulverized, mugwort leaves are in a fluffy state. Increasing the number of pulverizations, drying the mugwort leaves, and increasing the drying time only resulted in a maximum yield of 8.33% of the powder that could pass through the No. 6 sieve, which was far below 95.00%. Drying the mugwort leaves did not allow the powder to completely pass through the No. 2 sieve, and increasing the drying time did not significantly increase the yield of the powder that could pass through the No. 2 sieve, with a maximum of 96.79%. Without drying the mugwort leaves, pulverizing them twice allowed the powder to completely pass through the No. 2 sieve, and 34.66% of the powder could pass through the No. 4 sieve, reaching the coarse powder level.

[0034] (ii) Testing of the remaining medicinal materials after pulverization; Weigh out 103g of *Sargentodoxa cuneata*, 3g of *Homalomena occulta*, 3g of *Angelica pubescens*, 3g of *Dracaena cochinchinensis*, 3g of *Angelica dahurica*, 3g of *Carthamus tinctorius*, 3g of *Saposhnikovia divaricata*, 6g of *Viscum album*, 6g of *Boswellia carterii* (vinegar-processed), 6g of *Angelica sinensis*, 6g of *Dipsacus asper* (wine-processed), 6g of *Commiphora myrrha* (vinegar-processed), 6g of *Paeonia lactiflora*, 6g of *Periploca sepium*, 3g of *Zanthoxylum bungeanum* (fried), and 3g of *Notopterygium incisum*. Mix them in a stainless steel basin and then pulverize them in a graded ultrafine continuous water-cooled pulverizer equipped with a 100-mesh sieve. Weigh the powder, pass it through a No. 5 sieve, and weigh the powder that can pass through the sieve. Collect all the powder again, pass it through a No. 6 sieve, and weigh the powder that can pass through the sieve.

[0035] Sieveable powder yield: The sieveable powder yield was calculated. The results are shown in Table 2. ; Note: W—weight of the pulverized medicinal powder (g); W1—weight of the sieved medicinal powder (g).

[0036] Table 2: Results of the study on the yield of medicinal powder that can pass through different sieves

[0037] Experimental results show that after one pulverization, all sixteen other medicinal materials, including *Sargentodoxa cuneata*, can pass through a No. 5 sieve, and 96.94% of the powder can pass through a No. 6 sieve, reaching the fine powder grade.

[0038] (iii) Testing the mixing process of coarse Artemisia argyi powder and fine mixed powder; (1) Confirmation of indicator components; The content of paeoniflorin was used as an indicator for determining the uniformity of the mixture for the following reasons: 1. Paeoniflorin was not detected in *Sargentodoxa cuneata* and *Artemisia argyi*, which account for a relatively high proportion in the prescription. 2. According to the standard requirements for Paeonia lactiflora in the 2020 edition of the Chinese Pharmacopoeia, the content of paeoniflorin (C23H28O11) should not be less than 1.8%, which is relatively high, and the risk of detection sensitivity is low.

[0039] (2) Investigation on the analytical methods for paeoniflorin in powders; Referencing the method for determining paeoniflorin in the standard of Paeonia lactiflora in the 2020 edition of the Chinese Pharmacopoeia, Part I, we investigated the feasibility of this method.

[0040] Chromatographic conditions: Methanol-0.05 mol / L potassium dihydrogen phosphate solution (40:65) as mobile phase; detection wavelength: 230 nm; column: Kromasil 100-5-C18 (250 mm × 4.6 mm, 5 µm); injection volume: 10 μl; column temperature: 30 ℃; flow rate: 1.0 ml / min.

[0041] Blank solvent: methanol; Reference stock solution (0.5 mg / ml): Weigh 10 mg of paeoniflorin reference standard accurately, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with methanol, and shake well.

[0042] Negative control: Take about 0.43g of Artemisia argyi powder and about 0.34g of Sargentodoxa cuneata powder, weigh them accurately, put them in the same stoppered conical flask, add 20ml of methanol accurately, weigh them, soak for 4 hours, sonicate for 40 minutes, cool, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the negative control.

[0043] Test solution: Take about 1g of powder, accurately weigh it, place it in a stoppered conical flask, accurately add 20ml of methanol, weigh it, soak for 4 hours, sonicate for 40 minutes, cool, weigh it again, replenish the lost weight with methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0044] The results are as follows Figures 1 to 4 As shown, the minimum resolution between paeoniflorin and adjacent peaks in the test solution is 0.5, which is less than 1.5 and does not meet the requirements, making this method infeasible. Considering that the paeoniflorin peak in the test solution is too broad and adjusting the mobile phase ratio is not feasible, it is planned to change the type of mobile phase and increase the column temperature to reduce the peak broadening.

[0045] (3) Investigation on the analytical methods for paeoniflorin in powders (Part 2); The types of mobile phases used for the determination of paeoniflorin in the references were examined for feasibility; considering the complexity of the sample composition, a washing gradient was added; and the sample extraction time was also examined.

[0046] Chromatographic conditions: Column: Kromasil 100-5-C18 (250 mm × 4.6 mm, 5 µm); Mobile phase A: acetonitrile-0.1% phosphoric acid aqueous solution (20:80), Mobile phase B: acetonitrile, gradient elution (0–10 min 100% A, 10–11 min 100% A-10% A, 11–21 min 10% A, 21–30 min 100% A); Flow rate: 1.0 mL / min; Detection wavelength: 230 nm; Column temperature: 35 °C; Injection volume: 10 µL.

[0047] Extraction time investigation: Approximately 1g of the powder (batch number: 20230709) was accurately weighed and placed in a stoppered conical flask. 20ml of methanol was accurately added, and the mixture was weighed. The flask was soaked for 4 hours, then sonicated for 10, 20, and 30 minutes. After cooling, the mixture was weighed again. The lost weight was replenished with methanol, the mixture was shaken well, filtered, and the filtrate was collected. Two parallel preparations were performed and injected separately.

[0048] The results are as follows Figures 5 to 8 As shown, neither the blank solvent nor the negative control interfered with the elution of paeoniflorin. In the test solution, the resolution between paeoniflorin and adjacent peaks was at least 3.4 and greater than 1.5, meeting the requirements. Therefore, the chromatographic conditions were determined to be the chromatographic conditions for the detection of paeoniflorin in powders. When the ultrasonic time was 20 minutes and 30 minutes, the paeoniflorin content no longer increased, and the RD values ​​were both qualified (see Table 3). Considering the sample preparation efficiency, the ultrasonic time was set to 20 minutes, with other conditions remaining unchanged. This extraction method was determined to be the extraction method for paeoniflorin in powders.

[0049] Table 3: Ultrasonic Time Study of Test Samples

[0050] (4) Validation of the analytical method for paeoniflorin in powders; ① Specificity Blank solvent: methanol; Reference stock solution (0.5 mg / ml): Weigh approximately 10 mg of paeoniflorin reference standard accurately, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with methanol, and shake well.

[0051] Negative control: Take about 0.43g of Artemisia argyi powder and about 0.34g of Sargentodoxa cuneata powder, weigh them accurately, put them in the same stoppered conical flask, add 20ml of methanol accurately, weigh them, soak for 4 hours, sonicate for 20 minutes, cool, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the negative control.

[0052] Test solution: Take about 1g of powder, accurately weigh it, place it in a stoppered conical flask, accurately add 20ml of methanol, weigh it, soak for 4 hours, sonicate for 20 minutes, cool, weigh it again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0053] The above solution was measured under provisional chromatographic conditions.

[0054] Results: Neither the blank solvent nor the negative control interfered with the elution of paeoniflorin. In the test solution, the resolution between the paeoniflorin peak and its adjacent peak was at least 3.4, and greater than 1.5. The specificity of this method meets the requirements. The specificity overlay plot is shown in [Figure number missing]. Figure 2 .

[0055] ② Examination of linear relationships Reference solution (18 μg / ml): Accurately transfer 3.6 ml of the reference stock solution into a 100 ml volumetric flask, dilute to the mark with methanol, and mix well. Accurately pipette 1 μL, 2 μL, 5 μL, 8 μL, and 10 μL and determine the concentrations under the provisional chromatographic conditions. Plot the peak area as the ordinate (y) and the mass as the abscissa (x, ng), and perform linear regression to obtain the linear regression equation: y = 1.2457x + 20.19r 2 =0.9991, see Figure 3 As shown in the figure. The results indicate that the paeoniflorin mass has a good linear relationship with the peak area in the range of 107.5 ng to 1075.0 ng.

[0056] ③ Precision The reference solution was injected and analyzed under the provisional chromatographic conditions. The injection volume was 2 μl. The analysis was repeated 6 times, and the peak area was recorded. The RSD was 0.61% (n=6), indicating that the method has good precision. The results are shown in Table 4.

[0057] Table 4: Precision test of paeoniflorin

[0058] ④ Repeatability Six samples from the same batch (batch number 20230709) were taken and prepared according to the provisional test solution preparation method. The samples were injected and analyzed under the provisional chromatographic conditions. The peak area was recorded and the paeoniflorin content was calculated to be 0.537 mg / g. The RSD% was 1.57% (n=6), indicating that the method has good repeatability. The results are shown in Table 5.

[0059] Table 5: Repeatability test of paeoniflorin

[0060] ⑤ Stability test Take an appropriate amount of any test solution from the repeatability test, and inject it for analysis at room temperature for 0 h, 2 h, 6 h, 12 h, 18 h, and 24 h under the determined chromatographic conditions, recording the peak area. The results show that the change rate of the paeoniflorin peak area in the test solution after 24 h at room temperature is within 2%, indicating that paeoniflorin in the test solution is stable after 24 h at room temperature. The results are shown in Table 6.

[0061] Table 6: Stability test of paeoniflorin

[0062] ⑥ Accuracy test Approximately 1 g of sample (batch number 2023070901, paeoniflorin content 0.537 mg / g) was accurately weighed and divided into 6 portions. 1 mL of reference stock solution (0.5 mg / mL) was accurately added to each portion. Samples were prepared according to the provisional test solution preparation method and injected under provisional chromatographic conditions. The recovery rate was calculated. The average recovery rate of paeoniflorin was 101.1%, with an RSD of 4.06% (n=6), indicating good accuracy of the method. The results are shown in Table 7.

[0063] Table 7: Accuracy Test of Paeoniflorin

[0064] (5) Determined method for detecting paeoniflorin in powders Chromatographic column: Kromasil 100-5-C18 (250 mm × 4.6 mm, 5 µm); Mobile phase A: acetonitrile-0.1% phosphoric acid aqueous solution (20:80), mobile phase B: acetonitrile, gradient elution (0–10 min 100% A, 10–11 min 100% A–10% A, 11–21 min 10% A, 21–22 min 10% A–100% A, 22–30 min 100% A); Flow rate: 1.0 mL / min; Detection wavelength: 230 nm; Column temperature: 35 °C; Injection volume: 10 µL.

[0065] Extraction method for the test sample: Take about 1g of the powder, weigh it accurately, place it in a stoppered conical flask, add 20ml of methanol accurately, weigh it, soak for 4 hours, sonicate for 20 minutes, cool, weigh it again, make up the weight loss with methanol, shake well, filter, and take the filtrate to obtain the sample.

[0066] (6) Investigation of the mixing process Weigh out 3914g of *Sargentodoxa cuneata*, 114g of *Homalomena occulta*, 114g of *Angelica pubescens*, 114g of *Dracaena cochinchinensis*, 114g of *Angelica dahurica*, 114g of *Carthamus tinctorius*, 114g of *Saposhnikovia divaricata*, 228g of *Malva oleracea*, 228g of *Boswellia carterii* (vinegar-processed), 228g of *Angelica sinensis*, 228g of *Dipsacus asper* (wine-processed), 228g of *Commiphora myrrha* (vinegar-processed), 228g of *Paeonia lactiflora*, 228g of *Periploca sepium*, 228g of *Zanthoxylum bungeanum* (fried), and 228g of *Notopterygium incisum*. Mix them in a stainless steel basin, then pulverize them in a grading ultrafine continuous water-cooled pulverizer equipped with a 100-mesh sieve. Collect the powder to obtain *Sargentodoxa cuneata* mixed powder, weigh out 6529g of the powder, and set aside. Weigh out 5200g of *Artemisia argyi*, pulverize it in a grading ultrafine continuous water-cooled pulverizer equipped with a 100-mesh sieve, collect the powder, and pulverize it again in a grading ultrafine continuous water-cooled pulverizer equipped with a 100-mesh sieve to obtain *Artemisia argyi* powder, weigh out 5089g, and set aside.

[0067] The mixed powders of *Sargentodoxa cuneata* and *Artemisia argyi* were placed in a trough mixer (model: CH-200, manufacturer: Changzhou Xunchi Drying Equipment Co., Ltd.) and mixed. Approximately 1.5g of samples were taken at 5 different locations at 15, 30, and 45 minutes. The samples were prepared according to the test sample preparation method under (5) and the paeoniflorin content was determined. The results are shown in Table 8.

[0068] Table 8: Results of Paeoniflorin Content in Powders Obtained at Different Mixing Times (mg / g)

[0069] The experimental results showed that the paeoniflorin content tended to stabilize after 30 minutes of mixing, indicating that the powder was mixed evenly. Therefore, the mixing time was set at 45 minutes.

[0070] (iv) Screening test of powders; According to the proportions shown in the traditional Chinese medicine composition, Artemisia argyi powder (Experiment 3) was weighed and mixed with the powder of sixteen other medicinal materials such as Sargentodoxa cuneata. The yield of powder that could pass through a No. 2 sieve was then examined.

[0071] Sieveable powder yield: The yield of sieveable powder was calculated. The results are shown in Table 9; ; Note: W—weight of medicinal powder (g); W1—weight of sieveable medicinal powder (g).

[0072] Table 9: Results of the study on the yield of medicinal powder that can pass through a sieve

[0073] Experimental results show that the powder yield of the powder that can pass through the No. 2 sieve is 99.88%.

[0074] Furthermore, in order to examine the stability of the process, three batches of pilot-scale samples were prepared according to the optimized process, and the three batches of pilot-scale samples were tested according to the draft quality standard. The relevant process parameters and test results are shown in Table 10.

[0075] Table 10: Parameters and Test Results of Three Batches of Pilot Production

[0076] Furthermore, the results of the functional scores of patients treated with the traditional Chinese medicine composition provided in this application and the control group were compared and statistically analyzed, as shown in Table 11.

[0077] Table 11: Comparison of improvement in clinical symptoms and signs between the two groups

[0078] The treatment group received the external application powder described in this application, while the control group received Kangfu anti-inflammatory suppositories.

[0079] 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 traditional Chinese medicine composition for treating chronic pelvic inflammatory disease, characterized in that, The formula includes: 400-450g of Artemisia argyi, 10-50g of Angelica sinensis, 300-400g of Sargentodoxa cuneata, 10-50g of Dipsacus asper (processed with wine), 10-50g of Homalomena occulta, 10-50g of Commiphora myrrha (processed with vinegar), 10-50g of Angelica pubescens, 10-50g of Paeonia lactiflora, 10-50g of Daemonorops draco, 10-50g of Periploca sepium, 10-50g of Angelica dahurica, 10-50g of Saposhnikovia divaricata, 10-50g of Carthamus tinctorius, 10-50g of Viscum album, 10-50g of Zanthoxylum bungeanum (fried), 10-50g of Boswellia carterii (processed with vinegar), and 10-50g of Notopterygium incisum. in: Artemisia argyi and Sargentodoxa cuneata are the principal herbs; The main ingredients are safflower, frankincense, myrrh, red peony root, angelica root, and dragon's blood. The herbs used as adjuvants are Angelica pubescens, Notopterygium incisum, Loranthus parasiticus, Dipsacus asper, Periploca sepium, Homalomena occulta, Saposhnikovia divaricata, Angelica dahurica, and Zanthoxylum bungeanum.

2. The traditional Chinese medicine composition for treating chronic pelvic inflammatory disease according to claim 1, characterized in that, The formula includes 427g of Artemisia argyi, 20g of Angelica sinensis, 343g of Sargentodoxa cuneata, 20g of Dipsacus asper (processed with wine), 10g of Homalomena occulta, 20g of Myrrh (processed with vinegar), 10g of Angelica pubescens, 20g of Paeonia lactiflora, 10g of Dracaena cochinchinensis, 20g of Periploca sepium, 10g of Angelica dahurica, 20g of Saposhnikovia divaricata, 10g of Carthamus tinctorius, 20g of Viscum album, 10g of Zanthoxylum bungeanum (fried), 20g of Boswellia carterii (processed with vinegar), and 10g of Notopterygium incisum.

3. The traditional Chinese medicine composition for treating chronic pelvic inflammatory disease according to claim 1 or 2, characterized in that, The dosage form of the traditional Chinese medicine composition is: granules, capsules, or powder.

4. A method for preparing a traditional Chinese medicine composition for treating chronic pelvic inflammatory disease, characterized in that, Includes the following steps: Step 1: Take a portion of mugwort leaves and put them into a grading ultrafine continuous water-cooled pulverizer with a 100-mesh sieve for pulverization. After passing through a second sieve, you will get coarse mugwort powder. Take one part each of Angelica sinensis, Sargentodoxa cuneata, Dipsacus asper, Homalomena occulta, Myrrh, Angelica pubescens, Paeonia lactiflora, Daemonorops draco, Periploca sepium, Angelica dahurica, Saposhnikovia divaricata, Carthamus tinctorius, Mistletoe, Sichuan pepper, Boswellia carterii, and Notopterygium incisum, and put them into a grading ultrafine continuous water-cooled pulverizer with a 100-mesh sieve for pulverization. After passing through a No. 6 sieve, a mixed fine powder is obtained. Step 2: Mix the coarse powder of Artemisia argyi and the fine powder, and pass them through a No. 2 sieve to obtain a powder for treating chronic pelvic inflammatory disease.

5. The preparation method according to claim 4, characterized in that, The specific process for obtaining coarse mugwort powder is as follows: Take a portion of mugwort leaves and place them in a 100-mesh sieve for the first pulverization in a continuous water-cooled ultrafine pulverizer to obtain the first pulverized powder. The first-crushed powder is placed in a 100-mesh sieve and subjected to a second-crushing process in a continuous water-cooled ultrafine pulverizer to obtain the second-crushed powder. The powder was pulverized a second time and passed through a No. 2 sieve to obtain coarse Artemisia argyi powder.

6. The detection method for the powder for treating chronic pelvic inflammatory disease as described in claim 5, characterized in that, include: (i) The content of paeoniflorin was used as an indicator for determining the uniformity of mixing. (ii) Determination of paeoniflorin in Paeonia lactiflora according to the standard in Part I of the 2020 edition of the Chinese Pharmacopoeia; Chromatographic conditions: Column: Kromasil 100-5-C18; Mobile phase A: Acetonitrile-0.1% phosphoric acid aqueous solution; Mobile phase B: acetonitrile, gradient elution; Flow rate: 1.0 ml / min; Detection wavelength: 230nm; Column temperature: 35℃; Injection volume: 10 µl; Extraction time investigation: Take 1g of the powder and place it in a stoppered conical flask, add 20ml of methanol; weigh it and soak for 4 hours; sonicate for 10 minutes, 20 minutes or 30 minutes; cool it and weigh it again; make up the weight loss with methanol, shake well, filter, and take the filtrate to obtain the product; (III) Validation of the paeoniflorin analytical method; Blank solvent: methanol; 0.5 mg / ml reference stock solution: Weigh approximately 10 mg of paeoniflorin reference standard accurately, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with methanol, and shake well to obtain the solution. Negative control: Take about 0.43g of Artemisia argyi powder and about 0.34g of Sargentodoxa cuneata powder, weigh them accurately, put them in the same stoppered conical flask, add 20ml of methanol accurately, weigh them, soak for 4 hours, sonicate for 20 minutes, cool, weigh them again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the negative control. Test solution: Take about 1g of powder, weigh accurately, place it in a stoppered conical flask, add 20ml of methanol accurately, weigh, soak for 4 hours, sonicate for 20 minutes, cool, weigh again, replenish the lost weight with methanol, shake well, filter, and take the filtrate to obtain the test solution. The solution described in (iii) was determined according to the provisional chromatographic conditions; (iv) Examination of linear relationships; Reference solution 18 μg / ml: Accurately transfer 3.6 ml of the reference stock solution into a 100 ml volumetric flask, dilute to the mark with methanol, and shake well; (v) Precision; Take the reference solution and inject it into the chromatographic sample under the provisional chromatographic conditions. The injection volume is 2 μl. Repeat the analysis 6 times and record the peak area. (vi) Repetitiveness; Take 6 samples from the same batch, prepare test solutions according to the provisional test solution preparation method, inject and analyze according to the provisional chromatographic conditions, record the peak area, and calculate the paeoniflorin content; (vii) Stability test; Take an appropriate amount of any test solution under the repeatability test, and inject it for analysis at room temperature for 0h, 2h, 6h, 12h, 18h and 24h according to the determined chromatographic conditions, and record the peak area. (viii) Accuracy test; Take 1g of sample, accurately weigh it, and divide it into 6 portions. Add 1ml of reference stock solution to each portion accurately. Prepare the sample according to the provisional test solution preparation method, inject and determine the sample under the provisional chromatographic conditions, and calculate the recovery rate.

7. The detection method for the powder for treating chronic pelvic inflammatory disease according to claim 6, characterized in that, The ratio between acetonitrile and 0.1% phosphoric acid aqueous solution was set to 20:

80.

8. The detection method for the powder for treating chronic pelvic inflammatory disease according to claim 6, characterized in that, The specific process of gradient elution is as follows: 0~10min100%A, 10~11min100%A-10%A, 11~21min10%A, 21~30min100%A.

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