Auxiliary material composition prepared from rhizoma cyperi, rhizoma cyperi decoction pieces as well as preparation method and application of rhizoma cyperi decoction pieces

Abstract

The invention belongs to the technical field of traditional Chinese medicine processing, and particularly relates to an auxiliary material composition for processing rhizoma cyperi, rhizoma cyperi decoction pieces as well as a processing method and application of the rhizoma cyperi decoction pieces. The research finds that the rhizoma cyperi decoction piece prepared by combining auxiliary materials including fennel, folium artemisiae argyi, herba lycopi, herba elsholtziae, fructus alpiniae oxyphyllae and semen raphani shows an excellent effect in the aspect of treating liver depression, qi stagnation and blood stasis; the rhizoma cyperi decoction pieces prepared by combining auxiliary materials including fennel, folium artemisiae argyi, herba lycopi, herba elsholtziae, fructus alpiniae oxyphyllae, semen raphani, brown sugar, fresh ginger, table salt and table vinegar show an excellent effect in the aspect of treating primary dysmenorrhea and threatened abortion metrorrhagia. Besides, by optimizing processing technological parameters, the scientificity and stability of the processing process are ensured, the processing technology after auxiliary material combination optimization of the thirteen-processed rhizoma cyperi is preliminarily explored, and a basis is provided for modern industrial improvement of the thirteen-processed rhizoma cyperi.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine processing technology, specifically relating to the excipient composition for processing Cyperus rotundus, Cyperus rotundus slices, their processing methods and applications. Background Technology

[0002] Cyperus rotundus L., also known as sedge root, is the dried rhizome of the sedge plant (Cyperus rotundus L.) in the Cyperaceae family. It has the effects of soothing the liver and relieving depression, regulating menstruation and relieving pain, and regulating qi and the middle jiao. Clinically, it is often used to treat liver qi stagnation, chest and rib pain, indigestion, irregular menstruation, amenorrhea and dysmenorrhea, cold hernia abdominal pain, and breast distension. Cyperus rotundus was first recorded in the *Mingyi Bielu* (Records of Famous Physicians) and has been praised by traditional Chinese medicine practitioners throughout history as "the general manager of qi-related diseases and the chief of gynecology," making it a key and commonly used Chinese herb for regulating qi in clinical practice. Many ancient Chinese medical texts record different processing methods for Cyperus rotundus, and more than 20 processing methods are still used in various regions today. Different processing methods and auxiliary materials are often used for different indications, such as vinegar-processed, wine-processed, honey-processed, salt-processed, ginger-processed, four-processed Cyperus rotundus, and seven-processed Cyperus rotundus.

[0003] The processing of traditional Chinese medicine is influenced by regional culture, renowned medical schools, and folk experience, forming different academic systems, such as the four major processing schools of Sichuan, Beijing, Zhangzhou, and Jianchang, as well as other regional processing schools. In Chongqing, there is also a traditional thirteen-step method for processing Cyperus rotundus, a characteristic of the Sichuan school. This method consists of four steps, involving thirteen auxiliary materials (excluding water from the east wall): the first step is soaking the herbs in a thin mud-like mixture of soil from the east wall and glutinous rice straw ash; the second step is soaking them in children's urine; the third step is boiling them together with fennel seeds, mugwort leaves, Lycopus lucidus, Elsholtzia ciliata, Alpinia oxyphylla, and radish seeds; the fourth step is processing them with sugar, ginger, salt, and vinegar. This method was once widely used by the Tujia and Miao ethnic groups in southeastern Chongqing, mainly for postpartum lochia retention caused by qi stagnation and blood stasis in women. Previous research by our group has shown that the thirteen-processed Cyperus rotundus does indeed have a significant synergistic effect in improving blood rheology (whole blood viscosity, plasma viscosity, erythrocyte sedimentation rate, and hematocrit) and coagulation parameters, demonstrating great value in upholding tradition while innovating. However, the traditional thirteen-processed Cyperus rotundus processing system is extremely complex. Three of the four steps use compound excipients, and it also involves cold-prepared excipients such as Dongqiang soil and glutinous rice straw ash, as well as excipients with ethical controversies such as child urine, making the research extremely difficult.

[0004] Therefore, it is necessary to conduct in-depth research on the thirteen methods of processing Cyperus rotundus, simplify the processing techniques and improve its efficacy, so as to provide a basis for the modern industrial improvement of the thirteen-process Cyperus rotundus. Summary of the Invention

[0005] In view of this, one of the objectives of the present invention is to provide an excipient composition for processing Cyperus rotundus.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] An excipient composition for processing Cyperus rotundus, the excipient composition comprising the following components in parts by weight: 2-5 parts fennel, 2-5 parts artemisia argyi, 2-5 parts eupatorium fortunei, 2-5 parts elsholtzia ciliata, 2-5 parts Alpinia oxyphylla, and 2-5 parts radish seed.

[0008] Preferably, the excipient composition comprises the following components in parts by weight: 2.5 parts fennel, 2.5 parts mugwort, 2.5 parts eupatorium, 2.5 parts sesame, 2.5 parts alpinia oxyphylla, and 2.5 parts radish seed.

[0009] The second objective of this invention is to provide a Cyperus rotundus decoction piece.

[0010] To achieve the above objectives, the present invention adopts the following technical solution:

[0011] A type of Cyperus rotundus decoction piece is prepared by processing the aforementioned excipient composition to obtain Cyperus rotundus decoction piece.

[0012] The third objective of this invention is to provide a method for processing Cyperus rotundus using the aforementioned excipient composition.

[0013] To achieve the above objectives, the present invention adopts the following technical solution:

[0014] The method of processing Cyperus rotundus using the aforementioned excipient composition involves boiling Cyperus rotundus, the aforementioned excipient composition, and water together, and then drying to obtain the processed Cyperus rotundus.

[0015] Preferably, Cyperus rotundus is decocted while wrapped in gauze.

[0016] Preferably, the cooking time is 1 to 3 hours, and more preferably 3 hours.

[0017] Preferably, the drying temperature is 60~80℃, and more preferably 60℃.

[0018] Preferably, the mass ratio of Cyperus rotundus to the excipient composition is 100:15~25; the amount of water used is 8~14 times the total weight of Cyperus rotundus and the excipients.

[0019] Preferably, the mass ratio of Cyperus rotundus to the excipient composition is 100:15.

[0020] Preferably, the amount of water used is 14 times the total weight of Cyperus rotundus and the excipients.

[0021] The fourth objective of this invention is to provide an application of the aforementioned Cyperus rotundus decoction pieces in the preparation of a drug for treating liver qi stagnation and blood stasis syndrome.

[0022] Preferably, the treatment of liver qi stagnation and blood stasis includes reducing whole blood viscosity and plasma viscosity, reducing serum ALT and AST levels, and / or alleviating liver pathological morphology.

[0023] The fifth objective of this invention is to provide another excipient composition for processing Cyperus rotundus.

[0024] To achieve the above objectives, the present invention adopts the following technical solution:

[0025] Another excipient composition for processing Cyperus rotundus, the excipient composition comprising the following components in parts by weight: 2-5 parts fennel, 2-5 parts mugwort, 2-5 parts eupatorium, 2-5 parts elsholtzia, 2-5 parts alpinia oxyphylla, 2-5 parts radish seed, 8-12 parts brown sugar, 3-7 parts ginger, 1-3 parts salt, and 13-17 parts vinegar.

[0026] Preferably, the excipient composition comprises the following components in parts by weight: 2.5 parts fennel, 2.5 parts mugwort, 2.5 parts eupatorium, 2.5 parts sesame, 2.5 parts alpinia oxyphylla, 2.5 parts radish seed, 10 parts brown sugar, 5 parts ginger, 2 parts salt, and 15 parts vinegar.

[0027] The sixth objective of this invention is to provide another type of Cyperus rotundus decoction piece.

[0028] To achieve the above objectives, the present invention adopts the following technical solution:

[0029] Another type of Cyperus rotundus decoction piece is prepared by using the excipient composition described in Objective 5 to obtain Cyperus rotundus decoction piece.

[0030] The seventh objective of this invention is to provide a method for processing Cyperus rotundus using the excipient composition described in objective five.

[0031] To achieve the above objectives, the present invention adopts the following technical solution:

[0032] The method for processing Cyperus rotundus using the excipient composition described in Objective 5 involves processing Cyperus rotundus using the method described in Objective 3, adding a mixed solution of brown sugar, ginger, salt, and vinegar, mixing well, and then soaking and stir-frying to obtain processed Cyperus rotundus slices.

[0033] Preferably, the mass ratio of the Cyperus rotundus to the brown sugar, the ginger, the salt, and the vinegar is 80~120:8~12:3~7:1~3:13~17; and the weight ratio of the mixed solution of brown sugar, ginger, salt, and vinegar to the Cyperus rotundus is 0.5~0.7:1.

[0034] Preferably, the mass ratio of the Cyperus rotundus to the brown sugar, the ginger, the salt, and the vinegar is 100:10:5:2:15.

[0035] Preferably, the weight ratio of the mixed solution of brown sugar, ginger, salt, and vinegar to the Cyperus rotundus is 0.6:1.

[0036] Preferably, the mixed solution of brown sugar, ginger, salt, and vinegar is prepared by taking the prescribed amounts of brown sugar, ginger, salt, and vinegar, and adding an appropriate amount of water.

[0037] Preferably, the frying temperature is 150-170℃, the frying time is 15-35 minutes, and the frying frequency is 30-90 times / min.

[0038] Preferably, the stir-frying time is 25 minutes and the stirring frequency is 90 times / min.

[0039] Preferably, the solution is left to soak overnight.

[0040] The eighth objective of this invention is to provide the application of the Cyperus rotundus slices described in objective six in the preparation of a medicine for treating primary dysmenorrhea and / or uterine bleeding in threatened abortion.

[0041] Preferably, the treatment of primary dysmenorrhea includes reducing the body's uterine organ coefficient, alleviating pathological morphology, and / or reducing serum sex hormone levels.

[0042] Preferably, the reduction of pathological morphology includes reducing inflammatory infiltration of uterine tissue and uterine necrosis.

[0043] Preferably, the reduction of serum sex hormones includes reducing the levels of FSH, PGF2α, GnRH and / or LH.

[0044] Preferably, the treatment of threatened miscarriage uterine bleeding includes

[0045] Preferably, the treatment of threatened miscarriage uterine bleeding includes reducing vaginal bleeding and / or increasing serum sex hormone levels.

[0046] Preferably, the serum sex hormones include FSH and LH.

[0047] The beneficial effects of this invention are as follows:

[0048] 1. This study first focused on optimizing the excipients. Based on the characteristics of the traditional processing steps and excipient combinations, the study reduced the number of excipient combinations layer by layer according to the processing steps. Furthermore, it processed excipient combinations separately for different processing steps. Ultimately, seven combinations of excipients were designed for processing Cyperus rotundus, and seven processed samples of medicinal slices with different excipient combinations were prepared. By comparing the chemical composition transformation and efficacy of these seven processed samples, the study preliminarily explored the excipient optimization scheme for Cyperus rotundus, providing a basis for the modern industrial improvement of Cyperus rotundus.

[0049] 2. This invention, through detailed experimental research, verified the influence of different combinations of processing excipients on the efficacy of Cyperus rotundus. Experimental results show that Cyperus rotundus processed with a combination of excipients including fennel, mugwort, eupatorium, elsholtzia, alpinia oxyphylla, and radish seed exhibits excellent efficacy in treating liver qi stagnation and blood stasis; while Cyperus rotundus processed with a combination of excipients including fennel, mugwort, eupatorium, elsholtzia, alpinia oxyphylla, radish seed, brown sugar, ginger, salt, and vinegar shows excellent efficacy in treating primary dysmenorrhea and uterine bleeding from threatened abortion. This makes the processed Cyperus rotundus have broader application prospects and can meet different clinical needs.

[0050] 3. This invention improves the safety and operability of the processing method by optimizing the combination of excipients for processing Cyperus rotundus, removing excipients that are ethically controversial in traditional processing methods (such as children's urine) and excipients that are difficult to obtain (such as soil from the east wall and glutinous rice straw ash), making the processing method more in line with modern pharmaceutical standards.

[0051] 4. This invention optimizes process parameters such as water addition, cooking time, excipient ratio, and drying temperature to ensure the scientific nature and stability of the processing. Experimental results show that the optimized processing technology can significantly increase the content of extracts and volatile oils in Cyperus rotundus, thereby enhancing its efficacy. Attached Figure Description

[0052] Figures 1-10 The image shows the results of detecting the effects of seven processed Cyperus rotundus slices with different combinations of processing excipients on the pathological morphology of the uterus in a mouse model of primary dysmenorrhea (HE×200, scale bar=50 μm). Figure 1 HE staining image of uterine tissue from normal control mice; Figure 2 HE staining image of uterine tissue from mice in the model control group; Figure 3 HE staining image of uterine tissue from mice in the Yuanhu analgesic tablet group (0.01875 g / kg); Figure 4 HE staining image of uterine tissue from mice in group 1 (6.25 g / kg, Cyperus rotundus sample No. 1); Figure 5 HE staining image of uterine tissue from mice in the 6.25 g / kg group of Cyperus rotundus sample No. 2; Figure 6 HE staining image of uterine tissue from mice in the No. 3 Cyperus rotundus sample group (6.25 g / kg); Figure 7 HE staining image of uterine tissue from mice in the 6.25 g / kg group of Cyperus rotundus sample No. 4; Figure 8 HE staining image of uterine tissue from mice in the 6.25 g / kg group of Cyperus rotundus sample No. 5; Figure 9 HE staining image of uterine tissue from mice in the 6.25 g / kg group of Cyperus rotundus sample No. 6; Figure 10 HE staining image of uterine tissue from mice in the 6.25 g / kg group of Cyperus rotundus sample No. 7.

[0053] Figure 11 The images show the effects of seven processed Cyperus rotundus slices with different combinations of processing excipients on the pathological morphology of the liver in a mouse model of liver qi stagnation and blood stasis (HE×200, scale bar=50 μm). A represents HE staining of liver tissue from the normal control group; B represents HE staining of liver tissue from the model control group; C represents HE staining of liver tissue from the group treated with Jiawei Xiaoyao Wan (0.05 g / kg); D represents HE staining of liver tissue from the group treated with Cyperus rotundus sample 1 (6.25 g / kg); E represents HE staining of liver tissue from the group treated with Cyperus rotundus sample 2 (6.25 g / kg); F represents HE staining of liver tissue from the group treated with Cyperus rotundus sample 3 (6.25 g / kg); G represents HE staining of liver tissue from the group treated with Cyperus rotundus sample 4 (6.25 g / kg); H represents HE staining of liver tissue from the group treated with Cyperus rotundus sample 5 (6.25 g / kg); I represents HE staining of liver tissue from the group treated with Cyperus rotundus sample 6 (6.25 g / kg); J represents HE staining of liver tissue from the group treated with Cyperus rotundus sample 7 (6.25 g / kg). HE staining of liver tissue from mice in the g / kg group. Detailed Implementation

[0054] The technical solution of the present invention will be described more clearly and completely below with reference to specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Therefore, based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of the present invention.

[0055] Example 1

[0056] 1. Materials and Methods

[0057] 1.1 Test Drug

[0058] Raw Cyperus rotundus: Purchased from Sichuan Yifang Traditional Chinese Medicine Pieces Co., Ltd. (batch number: ZYC23030102, origin: Guangxi, specification: general). Identified by Chief Pharmacist Yang Min, Director of the National Traditional Chinese Medicine Processing Technology Inheritance Base of Chongqing Municipal Hospital of Traditional Chinese Medicine, as the dried rhizome of Cyperus rotundus L. (Cyperaceae family), it meets the requirements of the 2020 edition of the Chinese Pharmacopoeia. Seven processed Cyperus rotundus slices with different combinations of processing excipients were processed by the "National Traditional Chinese Medicine Processing Technology Inheritance Base (Chongqing)" of Chongqing Municipal Hospital of Traditional Chinese Medicine (processing methods and procedures are shown in Table 1), and randomly numbered 1-7 (samples processed individually with different processing steps and excipient combinations are numbered 1, 2, 3, and 7; samples processed with decreasing processing methods in the four steps are numbered 3, 4, 5, and 6, as detailed in Table 1).

[0059] Seven processed samples of Cyperus rotundus: Weigh 30 g of each of the seven processed samples of thirteen-processed Cyperus rotundus with different processing auxiliary materials combinations. After appropriate pulverization, transfer them to a round-bottom flask, and reflux extract with 10 times and 8 times the amount of water for 2 h respectively. After the extract is cooled, filter it by suction, centrifuge at 4000 r / min for 5 min, collect the supernatant, concentrate it under reduced pressure to 0.5 g of medicinal materials / mL, and store it refrigerated for later use.

[0060] Positive control drugs: Yuanhu Zhitong Tablets (Taiji Mianyang Pharmaceutical Co., Ltd., batch number: 2309008), Jiawei Xiaoyao Pills (Beijing Tongrentang Technology Development Co., Ltd., batch number: 240122), Gongxuening Capsules (Yunnan Baiyao Group Co., Ltd., batch number: 231205). After crushing, add them to water and dissolve, and prepare 75 mg / mL, 200 mg / mL and 7 mg / mL aqueous solutions respectively.

[0061] Tool drugs: Oxytocin Injection (Shanghai Quanyu Biotechnology Animal Pharmaceutical Co., Ltd., specification: 10 units / mL, batch number: 240207). Estradiol Benzoate Injection (Ningbo No. 2 Hormone Factory, specification: 2 mg / mL, batch number: C2209271). Mifepristone Tablets (China Resources Zizhu Pharmaceutical Co., Ltd., batch number: 240121); Misoprostol (China Resources Zizhu Pharmaceutical Co., Ltd., batch number: 240215). Take mifepristone tablets and misoprostol, after complete disintegration in water, stir evenly with a glass rod, and prepare suspensions of 0.85 mg / mL and 0.01 mg / mL respectively.

[0062] Table 1 Processing methods and process information table of seven samples of thirteen-processed Cyperus rotundus with different processing auxiliary materials combinations

[0063]

[0064] *Note: Sample No. 3 is not only the representative sample of the fourth-step processing auxiliary material combination in the different processing steps auxiliary material combinations, but also the representative sample of the last-step processing remaining after stripping the four processing steps layer by layer.

[0065] 1.2 Animals

[0066] SPF-grade female KM mice, weighing 18 - 22 g, were purchased from Chongqing Ensville Biotechnology Co., Ltd., production license number SCXK (Xiang) 2019 - 0004. The animals were raised in an environment with a 12 h light-dark cycle, a temperature of (22 ± 2) °C and a humidity of 50% ± 10%, with free access to water and food. After 3 d of adaptive feeding, the experiment began. This study complied with the relevant regulations on the management of experimental animals of Zunyi Medical University, and the experimental management and operation were carried out in accordance with the relevant regulations on national experimental animal ethics, in line with the animal ethics guiding principles and passed the approval (ethical approval number: ZMU21 - 2405 - 019).

[0067] 1.3 Reagents

[0068] Kits: Follicle-stimulating hormone (FSH, batch number: E-EL-M0511), prostaglandin 2α (PGF2α, batch number: E-EL-M1360), gonadotropin-releasing hormone (GnRH, batch number: E-EL-M0071), luteinizing hormone (LH, batch number: E-EL-M3053), alanine aminotransferase (ALT, batch number: E-BC-K235-M), and aspartate aminotransferase (AST, batch number: E-BC-K236-M) ELISA kits were all purchased from Elabscience Biotechnology Co., Ltd.

[0069] 1.4 Instruments

[0070] Inverted microscope (Olympus, Japan, model: BX43), low-temperature centrifuge (Eppendorf, Germany, model: 5425R), microplate reader (Thermo Scientific, USA, model: Varioskan LUX), electronic balance (Mettler-Toledo Instruments (Shanghai) Co., Ltd., model: ME204E), ultrapure water system (Chongqing Ashura Technology Development Co., Ltd., model: AXLM1820-2).

[0071] 1.5 Methods

[0072] 1.5.1 Comparison of therapeutic effects on primary dysmenorrhea in mice

[0073] 1.5.1.1 Experimental grouping and establishment of a primary dysmenorrhea model

[0074] One hundred mice were randomly divided into a normal control group, a model control group, a positive control group (using Yuanhu Zhitong tablets, 0.25 mL / 20 g, approximately 18.75 mg / kg), and a Cyperus rotundus sample group (0.25 mL / 20 g, approximately 6.25 g / kg), with 10 mice in each group. Except for the normal control group, the other mice were used to establish a mouse model of primary dysmenorrhea by combining estradiol benzoate and oxytocin. Specifically, each group of mice was intraperitoneally injected with estradiol benzoate (10 mL / kg) for 3 consecutive days. One hour before each injection, the mice were administered physiological saline (normal control group and model control group), aqueous extract of Cyperus rotundus sample (1-7), and aqueous solution of Yuanhu Zhitong tablets (positive control group) by gavage. On days 4 and 8, oxytocin (20 mL / kg) was injected intraperitoneally 30 minutes after the administration of the corresponding drugs by gavage. The writhing response within 30 minutes after the injection on day 8 was recorded. After the writhing was completed, blood was drawn through the orbital venous plexus, and the victim was then euthanized by cervical dislocation. The uterus was quickly removed, weighed, and then fixed in paraformaldehyde.

[0075] 1.5.1.2 Detection Indicators

[0076] 1.5.1.2.1 Detection of serum sex hormone levels in mice by ELISA

[0077] After weighing the mice in each experimental group, blood was collected from the orbital cavity of each mouse. The mice were centrifuged at 4000 r / min for 10 min, and the supernatant was collected. The serum levels of FSH, PGF2α, GnRH and LH were detected according to the ELISA kit instructions.

[0078] 1.5.1.2.2 Determination of Uterine Organ Coefficient

[0079] Mice in each experimental group were euthanized by cervical dislocation, and the abdominal cavity was opened to remove the uterus. The organ coefficient was then calculated. Uterine organ coefficient = total uterine mass (g) / body weight (g) × 100%.

[0080] 1.5.1.2.3 Uterine pathological morphology examination

[0081] A portion of uterine tissue was taken, fixed with tissue fixative, dehydrated with a gradient of ethanol, cleared with xylene, impregnated with paraffin, sectioned with paraffin, stained with hematoxylin and eosin (HE), dehydrated, cleared, mounted, and the pathological changes were observed under a microscope.

[0082] 1.5.2 Comparison of therapeutic effects on liver qi stagnation and blood stasis in mice

[0083] 1.5.2.1 Establishment of a mouse model of liver qi stagnation and blood stasis

[0084] A chronic unpredictable mild stress model was established. Methods included: ① water deprivation (24 h); ② tail clamping (1 min); ③ 45℃ environment (5 min); ④ reversed day-night cycle; ⑤ electric shock to the soles of the feet (50 mV, once every 50 s, each stimulation lasting 10 s, for a total of 30 times); ⑥ changes in living environment (damp bedding, tilted cage); ⑦ unfamiliar odors; ⑧ unfamiliar objects; ⑨ noise interference (1500 Hz, 95 dB, 1 h / d); ⑩ fasting for 24 h. These different stressors were applied throughout the experiment in a randomized order to prevent mice from anticipating the stimuli and thus avoid adaptation. Each stimulus was used twice on average. Mice were choked during the modeling process. Stressors were ensured to be unique within 3 days and continuously stimulated for 4 weeks. Successful modeling was indicated by the appearance of depressive-like behaviors in mice (significantly reduced sugar water preference, significantly reduced frequency of entering the central area of ​​the open field, and significantly prolonged immobility while swimming).

[0085] 1.5.2.2 Experimental grouping and drug administration

[0086] Mice that successfully developed the model were randomly divided into a model control group, a positive control group (using Jiawei Xiaoyao Wan, 0.25 mL / 20g, approximately 50 mg / kg of drug), and sample groups of Cyperus rotundus (0.25 mL / 20g, approximately 6.25 g / kg of raw drug), with 10 mice in each group, based on the aforementioned model evaluation indicators. Ten mice that did not undergo the modeling procedure were used as a normal control group. Modeling and drug administration were carried out simultaneously from day 1: the normal control group and model control group were administered physiological saline by gavage, the positive control group was administered Jiawei Xiaoyao Wan aqueous solution by gavage, and sample groups of Cyperus rotundus (1-7) were administered the corresponding Cyperus rotundus aqueous extract by gavage, twice daily for 14 consecutive days.

[0087] 1.5.2.3 Detection Indicators

[0088] 1.5.2.3.1 Mouse blood rheology detection

[0089] On day 15 of the experiment, blood was collected from the orbital cavity of mice in each experimental group. Whole blood anticoagulated with heparin sodium was used to measure the low shear (10 mPa / s), medium shear (50 mPa / s), and high shear (200 mPa / s) viscosity of whole blood and plasma viscosity on a fully automated blood rheometer. The state of "liver qi stagnation and blood stasis" in each experimental group was determined by evaluating the blood pathological state.

[0090] 1.5.2.3.2 Determination of ALT and AST content

[0091] Blood was collected from the orbital cavity of mice in each experimental group, centrifuged at 4000 r / min for 10 min, and the supernatant was collected. The serum ALT and AST levels were detected according to the ELISA kit instructions.

[0092] 1.5.2.3.3 Liver Pathological Morphology

[0093] After blood collection, the mice were euthanized, and liver tissue from each experimental group was removed. The tissue was washed with physiological saline, fixed with 4% paraformaldehyde, prepared into pathological sections, and stained with hematoxylin and eosin (HE). The lesions in the mouse liver tissue were observed under a microscope.

[0094] 1.5.3 Comparison of therapeutic effects on threatened abortion and uterine bleeding in mice

[0095] 1.5.3.1 Experimental grouping and establishment of a mouse model of threatened abortion and uterine hemorrhage

[0096] Female and male mice were caged at a ratio of 2:1. Vaginal plugs and vaginal smears were examined between 8:00 and 9:00 AM the following day; the presence of sperm was considered day 1 of pregnancy. One hundred pregnant mice were randomly selected and divided into a normal control group, a model control group, a positive control group (Gongxue Ning capsules, 0.25 mL / 20g, approximately 1.75 mg / kg), and sample groups of Cyperus rotundus (samples 1-17) (0.25 mL / 20g, approximately 6.25 g / kg of raw herb), with 10 mice in each group. Except for the normal control group, mice in the other groups were administered mifepristone (8.5 mg / kg) and misoprostol (100 μg / kg) by gavage to establish a mouse model of threatened abortion and uterine bleeding. The normal control group was administered an equal volume of physiological saline by gavage. On day 1 after successful model establishment, the normal control group and model control group were administered physiological saline by gavage, the positive control group was administered an aqueous solution of Gongxue Ning capsules by gavage, and the sample groups of Cyperus rotundus (samples 1-7) were administered an aqueous extract of Cyperus rotundus by gavage. Administration was continued for 7 days, twice daily. After the administration of the medication, a certain amount of sterile cotton balls (0.05 g in weight) were inserted into the vagina of the mice to absorb uterine bleeding. The cotton balls were removed at 8:00 AM on the second day, and a new cotton ball was placed in the vagina. At 5:00 PM, the blood-stained cotton ball was removed, and a new cotton ball was inserted. At the same time, the blood-stained cotton ball was placed in a sealed bag and refrigerated at 4 ℃. The same procedure was repeated daily until the 14th day after conception.

[0097] 1.5.3.2 Detection Indicators

[0098] 1.5.3.2.1 Vaginal bleeding volume in mice

[0099] Take 20 μL of venous blood from each experimental group of mice, add 4 mL of 5% NaOH solution, and mix well. Place the collected cotton balls containing blood from each mouse in a beaker, and add 5% NaOH solution in portions according to the amount of bleeding. Then soak, squeeze, and wash the cotton balls, soaking the cotton balls with less blood first, followed by the cotton balls with more blood. After soaking and extraction for 24 h, combine the extracts and shake well. Zero the instrument with 5% NaOH solution, and measure the absorbance (OD) of the extract and the NaOH solution containing 20 μL of mouse venous blood at 546 nm using a UV spectrophotometer. Simultaneously calculate the amount of uterine bleeding using the following formula: Uterine bleeding (mL) = Venous blood volume × (OD value of uterine extract × V2) / (OD value of venous blood × V1). Where V1) = the amount of NaOH solution used to dilute the venous blood (4 mL); V2 = the amount of NaOH solution used to extract uterine blood.

[0100] 1.5.3.2.2 Serum FSH and LH Level Detection

[0101] Blood was collected from the orbital sinus of mice in each experimental group, centrifuged at 4000 r / min for 10 min, and the supernatant was collected. The serum FSH and LH levels were detected according to the ELISA kit instructions.

[0102] 1.5.4 Statistical Processing

[0103] Statistical analysis was performed using SPSS 17.0 software. Serum sex hormone levels in each group showed a normal distribution as determined by a normality test. This indicates that one-way ANOVA was used to compare the sample means among multiple groups, and t-tests were used for pairwise comparisons between groups.

[0104] 2. Experimental Results

[0105] 2.1 Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on the number of writhing movements, uterine organ coefficients and pathological morphology, and serum sex hormone levels in a mouse model of primary dysmenorrhea.

[0106] 2.1.1 Number of writhing movements in mice

[0107] As shown in Table 2, compared with the normal control group, the number of writhing movements in the model control group mice was significantly increased (P<0.01). Compared with the model control group: (1) Among the four samples 3, 4, 5 and 6 that decreased in number, except for sample 5 which showed a significant decrease in the number of writhing movements (P<0.05), the other three samples showed a highly significant decrease in the number of writhing movements in mice (P<0.01); (2) Among the four samples 1, 2, 3 and 7 that were prepared separately, except for sample 1 which showed no statistical difference, the other three samples showed a highly significant decrease in the number of writhing movements in mice (P<0.01).

[0108] Table 2. Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on the number of writhing movements within 30 min in a mouse model of primary dysmenorrhea (n=10). )

[0109]

[0110] * Note: Significant differences exist. * P<0.05; the difference is highly significant. ** P<0.01.

[0111] 2.1.2 Uterine organ coefficient in mice

[0112] As shown in Table 3, compared with the normal control group, the uterine organ coefficient of the model control group mice was significantly increased (P<0.01). Compared with the model control group: (1) In the four samples 3, 4, 5 and 6, which decreased in number layer by layer, the uterine organ coefficient of mice was significantly reduced (P<0.05), among which sample 4 showed the most significant reduction (P<0.01); (2) In the four samples 1, 2, 3 and 7, which were processed separately, except for sample 1 which showed no statistical difference, the other three samples were significantly reduced (P<0.05), among which sample 2 showed the most significant reduction (P<0.01).

[0113] Table 3. Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on the uterine organ coefficient in a mouse model of primary dysmenorrhea (n=10, )

[0114]

[0115] * Note: Significant differences exist. * P<0.05; the difference is highly significant. ** P<0.01.

[0116] 2.1.3 Pathological morphology of mouse uterus

[0117] The results are as follows Figures 1-10 As shown. Compared with the normal control group, the uterus of mice in the model control group showed obvious neutrophil infiltration, uterine epithelial hyperplasia, edema, necrosis and shedding. Compared with the model control group: (1) Among the four samples 3, 4, 5 and 6 that decreased layer by layer, the uterine tissue of mice 3, 4 and 6 basically returned to normal, and the inflammatory infiltration and uterine necrosis were significantly reduced, while sample 5 showed a certain degree of reduction trend; (2) Among the four samples 1, 2, 3 and 7 that were prepared separately, only the uterine tissue of mice 3 and 7 basically returned to normal, and the inflammatory infiltration and uterine necrosis were reduced, while the others were not significantly different from the model group.

[0118] 2.1.4 Serum sex hormone levels in mice

[0119] As shown in Table 4, compared with the normal control group, the serum levels of FSH, PGF2α, GnRH and LH in the model control group mice were significantly increased (P<0.05). Compared with the model control group: (1) Among samples 3, 4, 5 and 6 with progressively reduced processing excipients, sample 6 significantly reduced FSH, PGF2α, GnRH and LH simultaneously (P<0.05), sample 3 significantly reduced FSH and PGF2α simultaneously (P<0.05), sample 4 significantly reduced GnRH and LH simultaneously (P<0.05), and sample 5 only significantly reduced LH (P<0.05); (2) Among samples 1, 2, 3 and 7 with separate processing, sample 7 significantly reduced PGF2α, GnRH and LH simultaneously (P<0.05), sample 2 significantly reduced PGF2α and GnRH simultaneously (P<0.05), sample 3 significantly reduced FSH and PGF2α simultaneously (P<0.05), and sample 1 had no significant regulatory effect on the above four indicators (P>0.05).

[0120] Table 4. Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on serum follicle-stimulating hormone (FSH), prostaglandin 2α (PGF2α), gonadotropin-releasing hormone (GnRH), and luteinizing hormone (LH) levels in a mouse model of primary dysmenorrhea (n=10, )

[0121]

[0122] * Note: Significant differences exist. * P<0.05; the difference is highly significant. ** P<0.01.

[0123] 2.2 Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on whole blood viscosity, serum ALT and AST levels, and liver pathological morphology in a mouse model of liver qi stagnation and blood stasis.

[0124] 2.2.1 Whole blood viscosity in mice

[0125] As shown in Table 5, compared with the normal control group, the whole blood viscosity and plasma viscosity of mice in the model control group were significantly increased (P<0.01). Compared with the model control group: (1) Among the four samples 3, 4, 5 and 6 that were progressively reduced, only samples 4 and 5 showed significantly decreased whole blood viscosity and plasma viscosity (P<0.05), while the other samples showed no significant changes; (2) Among the four samples 1, 2, 3 and 7 that were processed separately, only sample 2 showed significantly decreased whole blood viscosity and plasma viscosity (P<0.05), while the other samples showed no significant changes.

[0126] Table 5. Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on blood viscosity in a mouse model of liver qi stagnation and blood stasis (n=10). )

[0127]

[0128] * Note: Significant differences exist. * P<0.05; the difference is highly significant. ** P<0.01.

[0129] 2.2.2 Serum ALT and AST levels in mice

[0130] As shown in Table 6, compared with the normal control group, the serum ALT and AST levels of mice in the model control group were significantly increased (P<0.05). Compared with the model control group: (1) In samples 3, 4, 5, and 6 where the processing excipients were gradually reduced, the serum ALT and AST levels of mice were significantly decreased (P<0.05); (2) In samples 1, 2, 3, and 7 which were processed alone, samples 1, 2, and 3 significantly reduced the ALT and AST levels (P<0.05), while sample 7 had no significant regulatory effect on the above two indicators (P>0.05).

[0131] Table 6. Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in a mouse model of liver qi stagnation and blood stasis (n=10, )

[0132]

[0133] * Note: Significant differences exist. * P<0.05; the difference is highly significant. ** P<0.01.

[0134] 2.2.3 Pathological morphology of mouse liver

[0135] Compared with the normal control group, the model control group mice showed hepatocyte necrosis and mild hepatic sinusoidal dilation. Compared with the model control group, the liver lesions in mice treated with different processed Cyperus rotundus products all showed varying degrees of improvement. Among them, sample No. 5 (with progressively reduced quantities) and sample No. 2 (processed alone) showed the most significant improvement, with a significant reduction in the degree of hepatocyte necrosis, demonstrating the best therapeutic effect on liver qi stagnation and blood stasis. Figure 11 As shown.

[0136] 2.3 Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on vaginal bleeding and serum sex hormone levels in a mouse model of threatened abortion and uterine bleeding.

[0137] 2.3.1 Vaginal bleeding volume in mice

[0138] As shown in Table 7, compared with the normal control group, the amount of vaginal bleeding in the model control group mice was significantly increased (P<0.05). Compared with the model control group, the amount of vaginal bleeding in sample groups 4 and 6 was significantly reduced (P<0.05). There were no statistically significant differences between the remaining groups and the model group.

[0139] Table 7. Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on vaginal bleeding in a mouse model of threatened abortion and uterine bleeding (n=10). )

[0140]

[0141] * Note: Significant differences exist. * P<0.05; the difference is highly significant. ** P<0.01.

[0142] 2.3.2 Serum sex hormone levels in each group of mice

[0143] As shown in Table 8, compared with the normal control group, the serum FSH and LH levels of mice in the model control group were significantly reduced (P<0.05). Compared with the model control group: (1) Among samples 3, 4, 5, and 6 with progressively reduced processing excipients, samples 4, 5, and 6 significantly increased both FSH and LH levels (P<0.05), while sample 3 only significantly increased LH levels (P<0.05); (2) Among samples 1, 2, 3, and 7 with separate processing, only sample 2 significantly increased both FSH and LH levels (P<0.05), sample 3 significantly increased LH levels (P<0.05), sample 7 significantly increased FSH levels (P<0.05), and sample 1 had no significant regulatory effect on either of the above indicators (P>0.05).

[0144] Table 8. Effects of seven processed Cyperus rotundus samples with different combinations of processing excipients on serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels in a mouse model of threatened abortion and uterine bleeding (n=10). )

[0145]

[0146] Note: Compared with the model group, * P<0.05, ** P<0.01.

[0147] 3. Discussion

[0148] Cyperus rotundus can both soothe the liver and regulate qi, as well as invigorate blood and regulate menstruation. It has a significant regulating effect on menstrual problems in women caused by emotional distress, especially menstrual disorders and dysmenorrhea caused by liver qi stagnation. Modern research shows that the pharmacological effects of Cyperus rotundus are very extensive, but most of them are consistent with the traditional effects of "soothing the liver and relieving depression, regulating qi and relieving chest tightness, regulating menstruation and relieving pain". It is mainly concentrated in the fields of reproductive system, nervous system, digestive system, and cardiovascular system, such as regulating endocrine and improving menstrual disorders, inhibiting uterine smooth muscle contraction, improving polycystic ovary syndrome (PCOS), antidepressant and anti-anxiety, sedative and analgesic, regulating gastrointestinal motility, anti-gastric ulcer and gastric mucosal protection, choleretic effect, vasodilator and microcirculation improvement, anti-myocardial ischemia, and regulating blood lipids.

[0149] East wall soil, also known as "east wall soil," refers to the soil from the east wall of old houses. Because it receives the earliest sunlight each day, it is believed to have special medicinal value, primarily including detoxification, jaundice reduction, edema reduction, anti-inflammatory and antibacterial properties, and calming and soothing effects. This soil was readily available in ancient times because earthen walls still existed, but in modern times, earthen houses and walls have been almost entirely replaced by bricks and reinforced concrete, making it extremely difficult to find. Furthermore, modern processing standards almost entirely lack research on it. Rice straw ash, the ash produced by burning dried rice straw, has anti-inflammatory and heat-clearing detoxifying effects. While its source is slightly wider than east wall soil, it is also scarce due to the abandonment of land in modern rural areas, and modern processing standards rarely mention it. The above research shows that the auxiliary material combination of east wall soil and rice straw ash (auxiliary material combination A) does not significantly enhance the efficacy of Cyperus rotundus processing and can be considered for omission.

[0150] The use of children's urine in the processing of traditional Chinese medicine has a long history in my country, but it has also been highly controversial in modern times. One concern is whether it carries infectious agents, and another is the ethical obstacles it presents. In a previous summary of the historical evolution of Cyperus rotundus processing, the author found that among more than 60 processing methods, 45% involved children's urine. The usage rate reached 71.4% for two or more processing methods, and children's urine was always present in six or more processing methods. Therefore, researching the use of children's urine in the processing of Cyperus rotundus is a crucial issue. The aforementioned study showed that Cyperus rotundus processed with children's urine (excipient combination B) had a significant effect on treating primary dysmenorrhea in mice (P<0.05), but its effect on treating qi stagnation and blood stasis and threatened abortion uterine bleeding was not significant. Therefore, processing Cyperus rotundus with children's urine could be considered as one of the processing methods for treating dysmenorrhea in women, but the standardization of the use of children's urine as an excipient still needs further research.

[0151] The combination of fennel seeds, mugwort, eupatorium, senna, alpinia oxyphylla, and radish seeds is quite complex. Fennel seeds are pungent and warming, and can regulate and tonify the Chong and Ren meridians. Alpinia oxyphylla mainly treats deficiency and cold in the lower abdomen, assisting fennel seeds in warming the lower abdomen and uterus, and tonifying the liver and kidneys. Mugwort has the effects of promoting blood circulation, removing blood stasis, and clearing the meridians, promoting the discharge of blood stasis in the uterus and residual placental membranes; combined with eupatorium, it promotes blood circulation and eliminates blood stasis. Radish seeds are pungent, sweet, and neutral in nature, and enter the spleen, lung, and stomach meridians, guiding qi downward to promote the discharge of postpartum blood stasis from the uterus. Senna strengthens the body, tonifies deficiency, resolves phlegm, and regulates qi, and has a good regulatory effect on postpartum weakness and fatigue. The above studies show that when Cyperus rotundus is processed using a combination of excipients including fennel, mugwort, eupatorium, elsholtzia, alpinia oxyphylla, and radish seed (excipient combination C), it exhibits the best therapeutic effect on liver qi stagnation and blood stasis in mice (P<0.05). Furthermore, when combined with excipients such as slag, rice straw ash, children's urine, brown sugar, ginger, salt, and vinegar, it also shows significant therapeutic effects (P<0.05). In addition, when Cyperus rotundus is processed using a combination of excipients including fennel, mugwort, eupatorium, elsholtzia, alpinia oxyphylla, radish seed, brown sugar, ginger, salt, and vinegar (excipient combination CD), it also achieves the best therapeutic effect in mouse models of primary dysmenorrhea and threatened abortion with uterine bleeding (P<0.05). Therefore, it is possible to consider processing the following excipients (excipient combination C): fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, and radish seed, as an improved treatment for liver qi stagnation and blood stasis. Alternatively, processing the following excipients (excipient combination CD): fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, radish seed, brown sugar, ginger, salt, and vinegar, as an improved treatment for primary dysmenorrhea and threatened abortion uterine bleeding.

[0152] The four-processed Cyperus rotundus is one of the commonly used methods for processing Cyperus rotundus. Depending on the region and usage habits, the excipients vary. In Ningxia and Hubei, vinegar, brown sugar, wine, and salt are commonly used; in Henan, wine, milk, vinegar, and salt are commonly used; in Gansu, vinegar, wine, children's urine, and coarse salt were commonly used in the past; in Yunnan, white wine, vinegar, salt, and refined honey are commonly used; and in Jiangxi and Guangxi, vinegar, wine, salt, and ginger are commonly used. In Sichuan and Chongqing, vinegar, wine, salt, and ginger are also commonly used, but both yellow wine and white wine can be used. The four-processed method using brown sugar, ginger, salt, and vinegar is currently only reflected in the thirteen-processed Cyperus rotundus tradition. The above research shows that in the treatment of primary dysmenorrhea and threatened abortion uterine bleeding in mice, the excipient combination of fennel, mugwort, Lycopus lucidus, Elsholtzia ciliata, Alpinia oxyphylla, radish seed, brown sugar, ginger, salt, and vinegar (excipient combination CD) is the best processed product. Therefore, it is advisable to consider processing the following auxiliary ingredients (auxiliary ingredient combination CD) into a modified formula for treating primary dysmenorrhea and threatened abortion uterine bleeding: thirteen-processed Cyperus rotundus.

[0153] 4. Conclusion

[0154] In summary, the traditional Chongqing method of processing Cyperus rotundus significantly affects the main active ingredients and efficacy. It is worth considering modifying the original complex processing system by incorporating a combination of excipients (excipient combination C) of fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, and radish seed into a modified formula for treating liver qi stagnation and blood stasis. Furthermore, a combination of excipients (excipient combinations CD) of fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, radish seed, brown sugar, ginger, salt, and vinegar into a modified formula for treating primary dysmenorrhea and threatened abortion uterine bleeding. Further systematic research is needed on the excipient of child urine.

[0155] Example 2. Study on the processing technology of auxiliary material combination C

[0156] (1) Investigation of the amount of water added during cooking

[0157] Weigh out 7 portions each of Cyperus rotundus sample and excipients. Each portion contains 100g of Cyperus rotundus (wrapped in gauze for decoction) and 15g of excipients (15%, i.e., 2.5g each of fennel, mugwort, eupatorium, sesame, alpinia oxyphylla, and radish seed). Place them in a cooking container and add 6, 8, 10, 12, 14, 16, and 18 times the total amount of Cyperus rotundus and excipients of water respectively. Start timing after boiling. Check the remaining liquid every 1 hour of cooking. Stop heating when there is no liquid left.

[0158] The results are shown in Table 9. The cooking time and the amount of water added are generally directly proportional. The more water added, the longer the cooking time can be maintained.

[0159] Table 9. Examination of Water Addition for Cooking

[0160]

[0161] (2) Examination of cooking time

[0162] Weigh out 15 samples of Cyperus rotundus and 15 portions of excipients. Each portion contains 100 g of Cyperus rotundus (wrapped in gauze for decoction) and 15 g of excipients (15%, i.e., 2.5 g each of fennel, Artemisia argyi, Lycopus lucidus, Elsholtzia ciliata, Alpinia oxyphylla, and Raphanus sativus). Divide into 5 groups of 3 portions each. Based on the above-mentioned results on the amount of water added during decoction, add an appropriate amount of water and decoct for 1, 2, 3, 4, and 5 hours (h) respectively, ensuring a small amount of liquid remains. Remove the Cyperus rotundus and dry it. Determine the content of extract and volatile oil according to the Cyperus rotundus section of the 2025 edition of the Chinese Pharmacopoeia, calculate the average value of each group, and use a weighted comprehensive score to select the optimal decoction time. The weighted comprehensive score calculation formula is: F=a1×W1+a2×W2+…+a n ×W n Where F represents the comprehensive weighted score, a1-a n This represents the specific score of each indicator (a1=a1 / a) max ), W1-W nThe weights of each indicator are represented (in this experiment, the weights of volatile oil and extract content are each 50%).

[0163] The results are shown in Table 10-11. Different cooking times had different effects on the content of Cyperus rotundus extract and volatile oil. The optimal cooking time for auxiliary material combination C was 3 hours.

[0164] Table 10 Cooking Time Assessment Table

[0165]

[0166] Table 11 Weighted Overall Score Based on Cooking Time

[0167]

[0168] (3) Examination of the proportion of auxiliary materials

[0169] Weigh out 15 samples of Cyperus rotundus, each 100 g (wrapped in gauze and decocted), and make a group of 3 samples. Add 5 g (5%, i.e., 0.8 g each of fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, and radish seed) to each group, 10 g (10%, i.e., 1.7 g each of fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, and radish seed) to each group, 15 g (15%, i.e., 2.5 g each of fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, and radish seed) to each group, 20 g (20%, i.e., 3.3 g each of fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, and radish seed) to each group, and 25 g (25%, i.e., 4.4 g each of fennel, mugwort, eupatorium, sedge, alpinia oxyphylla, and radish seed) to each group, along with 14 times the total amount of water for Cyperus rotundus and the other auxiliary ingredients. Boil for 3 hours, then remove and dry the Cyperus rotundus. According to the 2025 edition of the Chinese Pharmacopoeia, the content of extract and volatile oil under Cyperus rotundus was determined, and the average value of each group was calculated. The optimal excipient ratio was selected by weighted comprehensive score.

[0170] The results are shown in Tables 12-13. Different amounts of excipients had different effects on the content of Cyperus rotundus extract and volatile oil. The optimal amount of excipient combination C was 15%.

[0171] Table 12 Analysis of Auxiliary Material Ratios

[0172]

[0173] Table 13 Weighted Overall Score Based on Cooking Time

[0174]

[0175] (4) Drying temperature investigation

[0176] Weigh out 6 portions each of Cyperus rotundus sample and excipients, 100 g of Cyperus rotundus per portion and 15 g of excipients per portion (i.e., 2.5 g each of fennel, Artemisia argyi, Lycopus lucidus, Elsholtzia ciliata, Alpinia oxyphylla, and Raphanus sativus seed). Take out 3 portions as a group, add 14 times the amount of water of Cyperus rotundus and excipients, boil for 3 hours, remove Cyperus rotundus, and dry at 60, 80 and 100 ℃ respectively. Determine the content of extract and volatile oil according to the Cyperus rotundus section of the 2025 edition of the Chinese Pharmacopoeia, calculate the average value, and screen the optimal drying temperature by weighted comprehensive score.

[0177] The results are shown in Tables 14-15. Different drying methods had different effects on the content of Cyperus rotundus extract and volatile oil. The optimal drying temperature for excipient combination C was 60 ℃.

[0178] Table 14 Drying Temperature Study

[0179]

[0180] Table 15 Weighted Comprehensive Score Based on Drying Temperature

[0181]

[0182] (5) Process validation

[0183] Based on the selected process conditions, three batches of samples were produced. The extract and volatile oil content were determined according to the Cyperus rotundus section of the 2025 edition of the Chinese Pharmacopoeia. The results of the three batches of samples were similar with small differences. Therefore, the selected excipient combination C process is considered to be scientific, stable, and feasible, and can be used as the process for processing Cyperus rotundus with excipient combination C. The results are shown in Table 16.

[0184] Table 16 Process Validation Experiment of Auxiliary Material Combination C

[0185]

[0186] In summary, the optimal processing method for excipient combination C is as follows: Take an appropriate amount of Cyperus rotundus, wrap it in gauze, place it in a cooking container, add an appropriate amount of excipient combination C, and add water at a ratio of 14 times the total weight of Cyperus rotundus and excipients. Cook for 3 hours, discard the excipients and dregs, remove the Cyperus rotundus, dry it at 60℃, and let it cool. For every 100 kg of Cyperus rotundus, use 15% excipient combination C (i.e., 2.5 kg each of fennel, Artemisia argyi, Lycopus lucidus, Elsholtzia ciliata, Alpinia oxyphylla, and Raphanus sativus seeds).

[0187] Example 3. Study on the processing technology of auxiliary material combination D

[0188] (1) Water absorption capacity assessment

[0189] Take 9 portions of Cyperus rotundus processed with excipient combination C, each portion weighing 100 g, and number them 1 to 9, and place them in beakers. Then take 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% of the weight of Cyperus rotundus and add them to samples 1 to 9 respectively, stirring constantly and letting them stand overnight to observe the water absorption.

[0190] The results are shown in Table 17. After processing with excipient combination C, the water absorption of Cyperus rotundus is about 60%. Therefore, the total amount of the moistening liquid processed with excipient combination D should be controlled at about 60% of the weight of Cyperus rotundus.

[0191] Table 17 Water Absorption Assessment Table

[0192]

[0193] (2) Investigation of the amount of excipients used

[0194] Select L9 (3) 4 An orthogonal experimental design was conducted using an orthogonal array, with the factor levels arranged as shown in Table 18. Eighteen portions of processed Cyperus rotundus (excipient combination C) were weighed, each portion weighing 100 g. Two portions were grouped together. The excipients were added according to the orthogonal design table, and the samples were allowed to soak and then dried to obtain the final samples. The extractive and volatile oil contents in the samples were determined according to the Cyperus rotundus section of the 2025 edition of the Chinese Pharmacopoeia, and a weighted comprehensive score was calculated. An analysis of variance was then performed on the weighted comprehensive score of the orthogonal experimental results.

[0195] The results are shown in Tables 19-20. Intuitive analysis shows that the influence of excipient dosage is as follows: Factor C (salt) > Factor A (brown sugar) > Factor B (ginger) > D (vinegar), with A1>A2>A3, B1>B3>B2, C2>C3>C1, and D3>D2>D1. Analysis of variance shows that Factor A (brown sugar), Factor B (ginger), and Factor C (salt) are extremely significant factors, while D (vinegar) is a significant factor. Therefore, considering the significance of the factors, the optimal level, and the actual application scenario, the optimal combination of excipient dosages is finally determined to be: A1B1C2D3, that is, for every 100 g of processed Cyperus rotundus using excipient combination C, use 10 g of brown sugar, 5 g of ginger, 2 g of salt, and 15 g of vinegar.

[0196] Table 18. Factor Level Table for Orthogonal Experiment of Auxiliary Material Usage

[0197]

[0198] Table 19 Experimental Results of Auxiliary Material Dosage

[0199]

[0200] Table 20 Weighted Composite Score ANOVA Table

[0201]

[0202] (3) Investigation of stir-frying process

[0203] Select L9 (3) 4 Orthogonal experimental design was performed using an orthogonal array, and the factor level arrangement is shown in Table 21. Nine portions of processed Cyperus rotundus (using excipient combination C) were weighed, each 100 g. A mixture of brown sugar, ginger, salt, and vinegar was added and mixed thoroughly, then left to soak overnight (for every 100 kg of processed Cyperus rotundus (using excipient combination C), 10 g of brown sugar, 5 g of ginger, 2 g of salt, and 15 g of vinegar were mixed with an appropriate amount of water; the weight ratio of the mixed solution to Cyperus rotundus was 0.6:1). The samples were then stir-fried according to the orthogonal design table to obtain the final samples. The extractives and volatile oil content in the Cyperus rotundus samples were determined according to the Cyperus rotundus section of the 2025 edition of the Chinese Pharmacopoeia, and a weighted comprehensive score was calculated. An analysis of variance was performed on the weighted comprehensive score of the orthogonal experimental results.

[0204] The results are shown in Tables 22-23. Intuitive analysis reveals that the degree of influence on the stir-frying process is as follows: Factor A (temperature) > Factor B (time) > Factor C (stirring frequency), with A2>A3>A1, B2>B3>B1, and C3>C2>C1. Analysis of variance shows that Factor A (temperature) is significant, while Factors B (time) and C (stirring frequency) have no significant impact. Therefore, considering the significance of the factors, the optimal level, and the actual application scenario, the optimal stir-frying process combination is finally determined to be: A2B2C3, i.e., stir-frying temperature 150-170℃, stir-frying time 25 min, and stir-frying frequency 90 times / min.

[0205] Table 21. Factor Level Table for Orthogonal Experiment of Stir-frying Process

[0206]

[0207] Table 22 Experimental Results of Stir-frying Process

[0208]

[0209] Table 23 Analysis of Variance for Stir-frying Process

[0210]

[0211] (4) Process validation

[0212] Based on the selected process conditions, three batches of samples were produced. The extractive and volatile oil contents in the samples were determined according to the Cyperus rotundus section of the 2025 edition of the Chinese Pharmacopoeia. The results of the three batches of samples were similar with small differences. It is believed that the selected excipient combination D process is scientific, stable, and feasible, and can be used as the process for processing Cyperus rotundus with excipient combination D. The results are shown in Table 24.

[0213] Table 24 Process Validation Experiment of Auxiliary Material Combination D

[0214]

[0215] In summary, the optimal processing method for auxiliary ingredient combination D is as follows: Take an appropriate amount of Cyperus rotundus processed from auxiliary ingredient combination C, add a mixture of brown sugar, ginger, salt, and vinegar, mix well, and let it soak overnight. Place it in a stir-frying container and stir-fry at 150-170 ℃ for 25 minutes, stirring 90 times / min. Remove and let cool. For every 100 kg of Cyperus rotundus processed from auxiliary ingredient combination C, use 10 g of brown sugar, 5 g of ginger, 2 g of salt, and 15 g of vinegar, along with an appropriate amount of water. The weight ratio of the mixed solution to Cyperus rotundus is 0.6:1.

Claims

1. An excipient composition for processing Rhizoma Cyperi, characterized in that, The adjuvant composition comprises the following components in parts by weight: cumin 2-5 parts, mugwort leaf 2-5 parts, privet 2-5 parts, mosla 2-5 parts, Chinese yam 2-5 parts, and radish seed 2-5 parts.

2. A Xiangfu decoction piece, characterized in that, The adjuvant composition of claim 1 is used for processing to obtain the prepared Xiangfu decoction pieces.

3. A method of processing Cyperus rotundus using the adjuvant composition according to claim 1, characterized in that, The Xiangfu, the adjuvant composition of claim 1, and clean water are boiled together, and dried to obtain the processed Xiangfu.

4. The method of claim 3, wherein, The boiling time is 1-3 hours.

5. The method of claim 3, wherein, The drying temperature is 60-80℃.

6. The method of claim 3, wherein, The mass ratio of the Xiangfu to the adjuvant composition is 100:15-25; and the amount of the clean water is 8-14 times the total weight of the Xiangfu and the adjuvant.

7. The use of the Xiangfu decoction pieces of claim 2 in the preparation of a medicament for treating liver stagnation and blood stasis.

8. Another composition of adjuvants for processing Rhizoma Cyperi, characterized in that, The adjuvant composition comprises the following components in parts by weight: cumin 2-5 parts, mugwort leaf 2-5 parts, privet 2-5 parts, mosla 2-5 parts, Chinese yam 2-5 parts, radish seed 2-5 parts, brown sugar 8-12 parts, ginger 3-7 parts, salt 1-3 parts, and vinegar 13-17 parts.

9. Another Rhizoma Cyperi decoction piece, characterized in that, The adjuvant composition of claim 8 is used for processing to obtain the Xiangfu decoction pieces.

10. A method of processing Rhizoma Cyperi with the adjuvant composition according to claim 8, characterized in that, The Xiangfu is processed by the method of any one of claims 3-6, mixed with a mixed solution of brown sugar, ginger, salt, and vinegar, and then moistened and fried to obtain the processed Xiangfu decoction pieces.

11. The method of claim 10, wherein, The mass ratio of the Xiangfu to the brown sugar, the ginger, the salt, and the vinegar is 80-120:8-12:3-7:1-3:13-17; and the weight ratio of the mixed solution of brown sugar, ginger, salt, and vinegar to the Xiangfu is 0.5-0.7:

1.

12. The method of claim 10, wherein, The frying temperature is 150-170℃, the frying time is 15-35 minutes, and the stirring frequency is 30-90 times per minute.

13. The use of the Xiangfu decoction pieces of claim 9 in the preparation of a medicament for treating primary dysmenorrhea and / or threatened abortion uterine bleeding.

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