Stasis-removing and pain-relieving pill and quality detection method thereof
By changing traditional Chinese medicine decoctions to pills and using starch paste as a binder, small granule pills were prepared, solving the problem of inconvenience in using traditional Chinese medicine decoctions and achieving the effects of portability and long-lasting efficacy.
Patent Information
- Application Number
- CN202511673239.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-14
- Publication Date
- 2026-02-24
Smart Images
Figure CN121550165A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a blood-activating and pain-relieving pill and its quality testing method. Background Technology
[0002] The Blood-Stasis-Expelling and Pain-Relieving Pill is derived from the Shaofu Blood-Stasis-Expelling Decoction with modifications. The entire formula consists of nine herbs: white peony root, corydalis rhizome, red peony root, angelica root, chuanxiong rhizome, cyperus rhizome, dried ginger, fennel seed, and astragalus root. This pill is designed to address the pathogenesis of this disease, which involves deficiency of qi and blood, and cold stagnation leading to blood stasis. In the formula, chuanxiong rhizome and angelica root are used together. Chuanxiong rhizome tonifies qi and generates blood, while also promoting qi and blood circulation, thus promoting the growth of yang and yin, and the flourishing of qi and blood. Together with angelica root, it tonifies blood deficiency and promotes blood circulation. Together, they serve as the principal herbs. White peony root and red peony root are used together to nourish and invigorate blood, soften the liver, and relieve pain. Dried ginger and fennel seed are used together to warm the meridians, dispel cold, and promote yang and unblock the channels. These four herbs serve as the assistant herbs. Cyperus rhizome and corydalis rhizome are used together to regulate qi, relieve depression, regulate menstruation, and relieve pain; these are all used as adjuvant herbs. When these herbs are combined, they work together to replenish and tonify, replenishing deficiencies in qi and blood, dispelling cold from the meridians, and promoting the flow of qi and blood, thus achieving the effects of "tonifying qi and blood, warming the meridians and dispelling cold, and removing blood stasis and relieving pain".
[0003] The Zhuyu Zhitong Pills have changed the dosage form of the Shaofu Zhuyu Decoction from a decoction to pills. The pills are small and lightweight, making them easy for patients to carry and take. Compared to decoctions, pills do not require decocting, reducing preparation time and inconvenience for patients. The slower release rate of pills allows the drug to act in the body for a longer period, resulting in a more sustained effect, making them suitable for long-term management of chronic diseases. The slow-release characteristic of pills reduces gastrointestinal irritation and adverse reactions. The portability and stability of pills make them more suitable for modern fast-paced lifestyles, especially for patients who frequently travel or require long-term medication, providing a more convenient option.
[0004] By studying the preparation process and quality standards of Zhuyu Zhitong Pills, we can gain a deeper understanding of the relevant theories and technologies of traditional Chinese medicine preparations, providing new ideas and methods for the development of traditional Chinese medicine preparations. It is expected to provide better options for common gynecological symptoms such as irregular menstruation, dysmenorrhea, and postpartum abdominal pain due to blood stasis, and alleviate the pain caused by these conditions. Summary of the Invention
[0005] In order to overcome the defects and deficiencies in the existing technology, the present invention provides a blood-activating and pain-relieving pill and its quality testing method.
[0006] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides a blood-activating and pain-relieving pill, wherein the raw materials used in the blood-activating and pain-relieving pill are composed of the following ingredients in parts by weight: 10-30 parts of white peony root, 10-30 parts of corydalis rhizome, 5-25 parts of red peony root, 10-30 parts of angelica root, 5-25 parts of chuanxiong rhizome, 5-25 parts of cyperus rhizome, 5-25 parts of dried ginger, 1-15 parts of fennel, and 10-30 parts of astragalus root. The binder used in the blood-activating and pain-relieving pill is 10%-15% starch paste in mass percentage.
[0007] As an optional method, in the above-mentioned blood-activating and pain-relieving pills, the raw materials used in the blood-activating and pain-relieving pills are composed of the following by weight: 20 parts of white peony root, 20 parts of corydalis rhizome, 15 parts of red peony root, 20 parts of angelica root, 15 parts of chuanxiong rhizome, 15 parts of cyperus rhizome, 15 parts of dried ginger, 10 parts of fennel, and 20 parts of astragalus root. The binder used in the blood-activating and pain-relieving pills is 10% starch paste by weight percentage.
[0008] In a second aspect, the present invention provides a method for preparing the blood-activating and pain-relieving pills described in the first aspect above, the method comprising the following steps: Mix an appropriate amount of raw drug powder with 10% starch paste by weight to form a soft material. Use an automatic pill-making machine for Chinese medicinal materials to make small granules with a diameter of 0.5~1 mm. Dry the prepared water pills to remove moisture and dry them at 60~80℃ for 8~12 hours to obtain the blood stasis relieving and pain-relieving pills.
[0009] Alternatively, in the above preparation method, 1530 g of raw drug powder is mixed with 1080 g of starch paste (10% by mass) to form a soft material. The soft material is then made into small particles with a diameter of 0.5-1 mm using an automatic pill-making machine for Chinese medicinal materials. The prepared water pills are then dried to remove moisture and dried at 60-80℃ for 10 hours to obtain the blood-activating and pain-relieving pills.
[0010] In a third aspect, the present invention provides a quality testing method for the blood-activating and pain-relieving pills described in the first aspect or prepared by the preparation method described in the second aspect, the quality testing method comprising: (1) General inspections: These include: disintegration degree test, moisture content test and weight difference test; (2) Microscopic identification; (3) Identification by thin-layer chromatography (TCL); and (4) High performance liquid chromatography (HPLC) content determination.
[0011] As an optional method, in the above quality testing methods, in (1) general inspection, the disintegration degree test adopts the disintegration time limit test method specified in the 2020 edition of the Chinese Pharmacopoeia, using a lifting disintegration apparatus, and the disintegration time of the Zhuyu Zhitong Pill is less than 8 minutes; the moisture content test adopts the drying method, and the moisture content of the Zhuyu Zhitong Pill is less than 5%; the weight difference test adopts the weight analysis test method specified in the 2020 edition of the Chinese Pharmacopoeia, and the weight difference of the Zhuyu Zhitong Pill is between 0.3% and 0.98%.
[0012] As an optional method, in the above quality testing method, in (2) microscopic identification, the powder of Zhuyu Zhitong Pill is light brown. According to the 2020 edition of the Chinese Pharmacopoeia, the powder of Zhuyu Zhitong Pill is identified by microscopic identification method.
[0013] As an optional approach, in the above quality inspection method, in (3) TCL identification, the TCL identification includes the following steps: (i) Preparation of the test solution: Take 6g of the sample of the Zhuyu Zhitong Pill, crush it, add ethyl acetate for ultrasonic extraction, centrifuge and filter, retain the filter residue, concentrate the filtrate to 2 mL by rotary evaporation, or evaporate it to dryness and then reconstitute it to 2 mL of ethyl acetate to obtain the solution. (ii) Preparation of reference herb solution: Take an appropriate amount of Angelica sinensis and Ligusticum chuanxiong reference herbs and prepare a reference herb solution using the same method as in (i); (iii) Prepare a double-negative pill without Angelica sinensis and Ligusticum chuanxiong according to the preparation method of Zhuyu Zhitong Pill. Weigh 6g of the pill and prepare a double-negative control solution without Angelica sinensis and Ligusticum chuanxiong in the same way as (i). (iv) According to the thin-layer chromatography method under General Chapter 0502 of Part IV of the 2020 edition of the Chinese Pharmacopoeia, take 5-8 μL of the test solution and the reference solution and spot them separately on the same silica gel G thin-layer plate. Use n-hexane:ethyl acetate volume ratio of 4:1 as the developing solvent. After development, remove and air dry, and observe under a 365 nm ultraviolet lamp. The results show that the chromatogram of the test sample and the reference sample show the same fluorescent spots at the same Rf value.
[0014] As an optional approach, in the above quality testing methods, in (3) TCL identification, in the Angelica sinensis-Ligusticum chuanxiong double negative system, the test sample and the control medicinal material both showed characteristic blue-green fluorescent spots at the same color development site Rf=0.69, and the Ligusticum chuanxiong negative control and the Angelica sinensis negative control obtained corresponding characteristic spots with Rf values of 0.66 and 0.68, respectively.
[0015] As an optional approach, in the above quality testing methods, in (4) HPLC content determination, the HPLC content determination includes the following steps: (a) Preparation of reference solution: Take an appropriate amount of ferulic acid reference standard, accurately weigh it, place it in a brown volumetric flask, add 70% methanol to prepare a solution, and you will get the solution. (b) Preparation of test solution: Take the powder of Zhuyu Zhitong Pill that has passed through No. 3 sieve, weigh it accurately and place it in a stoppered conical flask. Add an appropriate amount of 70% methanol accurately and quantitatively, turn on the constant temperature reflux device, process for 30 min, cool to room temperature and weigh it again, add 70% methanol to the initial total mass, let it stand and separate into layers, take the supernatant and filter it through a 0.45 μm microporous membrane, collect the filtrate for testing; (c) Chromatographic conditions: Column: C18 column, mobile phase: acetonitrile-0.085% phosphoric acid solution (volume ratio 17:83), flow rate: 1.0 mL / min, detection wavelength: 316 nm, column temperature: 35℃, theoretical plate number: ferulic acid peak ≥5000.
[0016] As an alternative, in the above quality detection methods, in (4) HPLC content determination, ferulic acid showed a good linear relationship in the range of 2~40 μg / mL.
[0017] Compared with the prior art, the beneficial effects of the present invention are: This invention provides new ideas and methods for the development of traditional Chinese medicine preparations through research on the preparation process and quality standards of Zhuyu Zhitong Pills. It is expected to provide a better option for common gynecological symptoms such as irregular menstruation, dysmenorrhea, and postpartum abdominal pain due to blood stasis, and alleviate the pain caused by these conditions. Attached Figure Description
[0018] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention, but do not constitute a limitation thereof. In the drawings: Figure 1 : Gelatinized starch - white peony root.
[0019] Figure 2 Calcium oxalate cluster crystals - white peony root.
[0020] Figure 3 : Hypothenar thick-walled cells - Corydalis rhizome.
[0021] Figure 4 Stone cells - Corydalis rhizome.
[0022] Figure 5 Starch granules - Red peony root.
[0023] Figure 6 : Spindle-shaped phloem parenchyma cells - Angelica sinensis.
[0024] Figure 7 : Oil chamber - Angelica sinensis.
[0025] Figure 8Calcium oxalate crystals - Ligusticum chuanxiong.
[0026] Figure 9 Secretory cells - Cyperus rotundus.
[0027] Figure 10 Hypodermic fibers - Cyperus rotundus.
[0028] Figure 11 Oil cells - dried ginger.
[0029] Figure 12 Oil cell fragments - fennel seeds.
[0030] Figure 13 : Endocarp mosaic cells - fennel.
[0031] Figure 14 : Fiber - Astragalus.
[0032] Figure 15 TCL chromatogram of Zhuyu Zhitong Pill (Danggui and Chuanxiong double negative). Among them, 1, 2, and 3 are Zhuyu Zhitong Pill test sample; 4 is Danggui reference standard; 5 is Chuanxiong reference standard; 6 is negative preparation.
[0033] Figure 16 TCL chromatogram of Zhuyu Zhitong Pill (Chuanxiong). Among them, 1, 2, and 3 are Zhuyu Zhitong Pill test sample; 4 is Chuanxiong reference standard; and 5 is negative control preparation.
[0034] Figure 17 TCL chromatogram of Zhuyu Zhitong Pill (Angelica sinensis). Among them, 1, 2, and 3 are Zhuyu Zhitong Pill test sample; 4 is Angelica sinensis reference standard; and 5 is negative control preparation.
[0035] Figure 18 : Specificity examination.
[0036] Figure 19 : Examining linear relationships.
[0037] Figure 20 : Regression curve.
[0038] Figure 21 Repeatability determination. Detailed Implementation
[0039] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Unless otherwise specified, the materials and reagents used are commercially available.
[0040] Example 1: Study on the preparation method of Zhuyu Zhitong Pill 1. Prescription confirmation: The blood-stasis-removing and pain-relieving pills are made from the following raw materials: 20 g of white peony root, 20 g of corydalis rhizome, 15 g of red peony root, 20 g of angelica root, 15 g of chuanxiong rhizome, 15 g of cyperus rhizome, 15 g of dried ginger, 10 g of fennel, and 20 g of astragalus root.
[0041] 2. Making pellets 2.1 Process Screening The molding process of Zhuyu Zhitong Pills focused on a comparative screening of starch paste and dextrin. Using molding rate, disintegration time, and pill weight difference as indicators, single-factor experiments determined the following: The starch paste group, using a 10% concentration, mixed the medicinal powder and paste in a 7:3 ratio, and prepared water pills using a molding method, achieved a molding rate of 92.3% and a disintegration time of 6 minutes. The dextrin group, using a 20% dextrin solution as a binder, with a medicinal powder to dextrin solution ratio of 6:4, achieved a molding rate of 90.1%, but a disintegration time of 78 minutes, exceeding the pharmacopoeia standard. Considering all factors, starch paste was selected as the binder.
[0042] 2.2 Screening of Starch Paste A total of 2900 g of the raw materials in the prescription were dried in a forced-air drying oven at 50-60℃ for 12 hours, then removed, pulverized, passed through a 100-mesh sieve, and mixed evenly to obtain 2340 g of fine powder, with a powder yield of 81%. Starch paste was selected as a binder for the preparation of the blood-activating and pain-relieving pills. This experiment used 8%, 10%, 12%, and 15% starch paste. See Table 1.
[0043] Table 1. Investigation of starch paste ratio In conclusion, 10% starch paste was selected as the binder in this formula.
[0044] 2.3 Results 1530 g of medicinal powder was mixed with 1080 g of 10% starch slurry to form a soft mass. This soft mass was then granulated into small pellets with a diameter of 0.5–1 mm using an automatic pill-making machine for traditional Chinese medicine. The resulting pills were dried at 60–80°C for 10 hours to remove moisture. After drying, the pills weighed approximately 0.6 g each, exhibited good disintegration, and had a moisture content of approximately 3.9%.
[0045] The Zhuyu Zhitong Pill has changed the dosage form of the Shaofu Zhuyu Decoction from a decoction to a pill. The pill is small in size and light in weight, making it easy for patients to carry and take, and is especially suitable for patients who need long-term medication.
[0046] Example 2: Quality Testing Method for Zhuyu Zhitong Pills 1. General inspection 1.1 Determination of disintegration The disintegration degree of Zhuyu Zhitong Pills can be determined by the disintegration time limit test method specified in the 2020 edition of the Chinese Pharmacopoeia, using a lifting disintegration apparatus.
[0047] Take 6 test pellets and place them separately in a special glass tube for disintegration baskets. Place a stainless steel baffle with a 2.0 mm aperture at the bottom of each glass tube to ensure that the pellets are not suspended in the air at the contact surface with the baffle. All pellets should dissolve or the disintegrating material should pass through the baffle mesh within 30 minutes. If ≥1 pellet fails to disintegrate completely in the initial test of 6 pellets, another 6 pellets should be taken for retesting, and all retests should meet the requirements. See Table 2.
[0048] Table 2. Disintegration time of Zhuyu Zhitong Pills Results analysis: Based on the experimental data of the three groups, the disintegration time of Zhuyu Zhitong Pills in each group was within the prescribed 30 minutes, and the average values were 7.3 minutes, 6 minutes and 5.8 minutes, respectively, all of which were much less than the prescribed 30 minutes.
[0049] In summary, the disintegration degree of the tested sample meets the requirements, indicating that the Zhuyu Zhitong Pill has good disintegration performance within the specified time.
[0050] 1.2 Moisture Content Determination Take 2-5g of the sample to be tested, crush it to ensure the particle size is ≤3 mm, and spread it evenly in a constant-weight flat weighing bottle, controlling the thickness to ≤5 mm. Open the bottle cap and dry in a drying oven at 100-105℃ for 5 hours initially. After capping, transfer it to a desiccator to cool to room temperature for 30 minutes, and accurately weigh it (accuracy ≤0.1 mg). Repeat the drying-cooling-weighing process for 1 hour. The termination condition is: the difference between two consecutive weighings ≤5 mg. See Table 3.
[0051] Table 3 Moisture Content Measurement Results analysis: The moisture contents of the three groups of samples were 4.61%, 3.70%, and 3.43%, respectively. With the calculated average moisture content M = 3.91% as a reference, it indicates that the overall moisture content of this product fluctuates between approximately 3.43% and 4.61%, with an average moisture content of 3.91%.
[0052] 1.3 Weight Difference Ten pills of the test sample were taken as one part, and ten parts were weighed and compared with the average weight. See Table 4.
[0053] Table 4. Weight Differences of Zhuyu Zhitong Pills Results Analysis: The weight variation data of the 10 test samples showed that the weight differences among the various samples of Zhuyu Zhitong Pills were small, all at relatively low levels. The weight variation of most samples was concentrated between 0.3% and 0.98%, indicating that the weight of this batch of Zhuyu Zhitong Pills was relatively uniform, and the production process provided stable control over the weight of the pills.
[0054] 2. Microscopic identification The powder of this product is light brown. Take three small batches of powder, mount them on dilute glycerin, add chloral hydrate test solution, and slowly heat over an alcohol lamp to allow it to permeate. Pay attention to the temperature and time to avoid carbonization or over-permeation of the sample. After permeation, add dilute glycerin to seal the slide, avoiding air bubbles when adding the coverslip. Identify the powder of this product using a microscopic identification method according to the 2020 edition of the Chinese Pharmacopoeia.
[0055] 2.1 Characteristics White peony root: The surface is slightly yellow or light brown, and some may have scorched spots. It has a slightly fragrant aroma.
[0056] Corydalis: This product is irregularly flattened spherical, with a yellow or yellowish-brown surface and bumpy protrusions at the bottom.
[0057] Red peony: This product is cylindrical, with a brownish-brown, rough surface, and longitudinal grooves and wrinkles.
[0058] Angelica sinensis: The cut surface is slightly yellow or light brown, and some may show charred spots. It has a slight aroma of wine.
[0059] Chuanxiong: This product is an irregular, nodular, fist-shaped mass with a grayish-brown or brown surface that is rough and wrinkled.
[0060] Cyperus rotundus: The surface is dark brown with a slight vinegar aroma.
[0061] Dried ginger: This product is in the form of flat, blocky pieces with finger-like branches, and its surface is grayish-yellow or light grayish-brown.
[0062] Fennel seeds: slightly swollen, dark in color, and slightly salty in taste.
[0063] Astragalus: This product is cylindrical, with a light brown surface and irregular longitudinal grooves and wrinkles.
[0064] 2.2 Microscopic identification of each active pharmaceutical ingredient White peony root: Microscopically, numerous gelatinized starch granules filling the parenchyma cells are visible; simultaneously, numerous calcium oxalate clusters arranged in multiple rows are also visible, scattered or present in the parenchyma cells. See Figure 1 and Figure 2 .
[0065] Corydalis: Fibers show yellowish-green, thick-walled hypodermal cells with lignified walls, some exhibiting beaded thickening, and fine pits. See [link to original text]. Figure 3 and Figure 4 .
[0066] Red peony root: Microscopically, calcium oxalate clusters are scattered or present in thin-walled cells, often arranged in rows. Numerous starch granules are also visible; single granules are oval, and compound granules consist of 2-4 subgranules. See [link to article]. Figure 5 .
[0067] Angelica sinensis: Microscopically, spindle-shaped phloem parenchyma cells with extremely fine, obliquely interwoven reticulate textures and oil cell fragments containing yellowish-brown secretions are visible. See Figure 6 and Figure 7 .
[0068] Chuanxiong (Ligusticum striatum): Microscopically visible are round or irregularly shaped calcium oxalate crystals. See... Figure 8 .
[0069] Cyperus rotundus: Microscopically, round or polygonal secretory cells containing yellowish-brown or reddish-brown secretions are visible. See Figure 9 and Figure 10 .
[0070] Dried ginger: Microscopic examination reveals scattered oil cells and resin cells within the parenchyma, their lumens containing pale yellow lipid droplets or brownish-red resinous substances. See [link to article]. Figure 11 .
[0071] Fennel: Microscopically, irregularly mosaicked endocarp cells are visible; surface view shows elongated, slightly wavy cells, often arranged in groups of several in an irregular mosaic pattern; yellowish-brown oil cell fragments are also observed; secretory cells are polygonal and contain yellowish-brown secretions. See Figure 12 and Figure 13 .
[0072] Astragalus: Microscopically, the fiber bundles are longitudinally split at the broken ends into a broom-like shape; they are slender, colorless, with extremely thick walls and indistinct pits. See [link to article]. Figure 14 .
[0073] 3. TCL identification 3.1 Preparation of the test solution Take 6 g of sample, crush it, add 20 mL of ethyl acetate and extract it by ultrasonication for 20 min, centrifuge and filter, and retain the filter residue; concentrate the filtrate to 2 mL by rotary evaporation, or evaporate it to dryness and then reconstitute it to 2 mL of ethyl acetate to obtain the final product.
[0074] 3.2 Preparation of control herbal solution Take 1 g each of Angelica sinensis and Ligusticum chuanxiong as reference herbs and prepare a reference herb solution using the same method.
[0075] 3.3 Preparation of negative control solution Weigh out 20 parts of white peony root (excluding angelica root and chuanxiong rhizome), 20 parts of corydalis rhizome, 15 parts of red peony root, 15 parts of cyperus rhizome, 15 parts of dried ginger, 10 parts of fennel seed, and 20 parts of astragalus root. Using 10% starch paste as a binder, prepare a double-negative control solution (excluding angelica root and chuanxiong rhizome) according to the preparation method of Zhuyu Zhitong Pill. Weigh out 6g of the above pills and prepare a double-negative control solution (excluding angelica root and chuanxiong rhizome) according to the preparation method of test solution in 3.1.
[0076] 3.4 Thin-layer chromatography Thin-layer chromatography (TLC) was performed according to Section 0502 of the General Chapter IV of the 2020 edition of the Chinese Pharmacopoeia. 5-8 μL each of the test solution and the control solution were spotted separately onto the same silica gel G TLC plate. Hexane-ethyl acetate (v / v: 4:1) was used as the developing solvent. After development, the plate was removed, air-dried, and observed under a 365 nm UV lamp. The results showed that the test sample and the control solution exhibited fluorescent spots of the same color at the same Rf value. (See attached table for details.) Figure 15 , Figure 16 and Figure 17 .
[0077] 3.5 Results Analysis In the Angelica sinensis-Ligusticum chuanxiong double-negative system ( Figure 15 Both the test sample and the control sample exhibited characteristic blue-green fluorescent spots at the same chromatographic site (Rf=0.69), while the green spots in the negative control group showed significantly different chromatographic behavior from these characteristic spots. (Ligusticum chuanxiong negative...) Figure 16 ) and Angelica sinensis (yin) Figure 17 The experiments yielded characteristic spots with Rf values of 0.66 and 0.68, respectively, and no specific interference signals were observed in the negative control systems. In all three experiments, the Rf values of the characteristic spots remained stable in the range of 0.66-0.69, and the chromatographic behavior of each negative control group maintained sufficient separation from the target component.
[0078] The established chromatographic conditions effectively distinguished the target component from matrix interferences during specificity validation, and the method validation parameters confirmed that the detection system has good chromatographic separation efficiency. It has high specificity (no false positive interference) and meets the quality control requirements for the identification and analysis of components in traditional Chinese medicine.
[0079] 4. HPLC content determination 4.1 Chromatographic conditions Chromatographic column: Elite C18 column (250 mm × 4.6 mm, 5 μm), mobile phase: acetonitrile-0.085% phosphoric acid solution (volume ratio 17:83), flow rate: 1.0 mL / min, detection wavelength: 316 nm, column temperature: 35℃, theoretical plate number: ferulic acid peak ≥5000.
[0080] 4.2 Preparation of reference solution Take an appropriate amount of ferulic acid reference standard, accurately weigh it, place it in a brown volumetric flask, and add 70% methanol to prepare a solution containing 12 μg per 1 mL.
[0081] 4.3 Preparation of the test solution Take (2.00±0.05) g of sample powder through a No. 3 sieve, accurately weigh it, and place it in a stoppered conical flask. Accurately add 20 mL of 70% methanol, seal the flask tightly, and double-weigh for calibration. Turn on the constant temperature reflux device and maintain the extraction temperature at (80±5)℃ for 30 min. After cooling to room temperature, weigh it again, add 70% methanol to the initial total mass, let it stand and separate into layers, and filter the supernatant through a 0.45 μm microporous membrane. Collect the filtrate for analysis.
[0082] 4.4 Specificity Examination Under the above chromatographic conditions, place the test solution and reference solution into the HPLC chromatograph, set the program for automatic injection, inject 10 µL each time, and determine the peak areas as shown in Table 6. Record the chromatograms as shown in Table 6. Figure 18 .
[0083] Table 6 Specificity Results according to Figure 18 As shown in Table 6, the retention time of ferulic acid peaks is similar and has no effect on its determination, indicating that the experimental method is feasible.
[0084] 4.5 Examination of Linear Relationships Five different volumes of the reference solution were prepared to concentrations of 5, 10, 20, 30, and 40 μg / mL. One syringe was used for each sample, and detection was performed at a wavelength of 316 nm. The results are as follows. Figure 19 and Figure 20 Table 7.
[0085] Table 7 Linear regression curves of ferulic acid Results analysis: y = 0.5736x - 0.2351, R² = 0.9997, ferulic acid showed a good linear relationship in the range of 2~40 μg / mL.
[0086] 4.6 Repeatability determination Take 10 μL of each of the six batches of test solutions and detect them at a wavelength of 316 nm. The results are shown in Table 8 below. Figure 21 .
[0087] Table 8 Results of Repeatability Tests Results analysis: The peak area RSD was 1.36% and the content RSD was 0.41% in the six determinations, both of which met the requirements (Pharmacopoeia 9101).
[0088] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several modifications or equivalent substitutions can be made to the technical solution without departing from the principle of the present invention, and these modifications or equivalent substitutions should also be considered within the scope of protection of the present invention.
Claims
1. A blood-activating and pain-relieving pill, characterized in that: By weight, the raw materials used in the blood-activating and pain-relieving pills consist of the following: 10-30 parts of white peony root, 10-30 parts of corydalis rhizome, 5-25 parts of red peony root, 10-30 parts of angelica root, 5-25 parts of chuanxiong rhizome, 5-25 parts of cyperus rhizome, 5-25 parts of dried ginger, 1-15 parts of fennel, and 10-30 parts of astragalus root. By mass percentage, the binder used in the blood-activating and pain-relieving pills is 10%-15% starch paste.
2. The blood-activating and pain-relieving pill according to claim 1, characterized in that: By weight, the raw materials used in the blood-activating and pain-relieving pills consist of the following: 20 parts of white peony root, 20 parts of corydalis rhizome, 15 parts of red peony root, 20 parts of angelica root, 15 parts of chuanxiong rhizome, 15 parts of cyperus rhizome, 15 parts of dried ginger, 10 parts of fennel, and 20 parts of astragalus root. By mass percentage, the binder used in the blood-activating and pain-relieving pills is 10% starch paste.
3. The preparation method of the blood-activating and pain-relieving pill according to claim 1 or claim 2, characterized in that: The preparation method includes the following steps: Mix an appropriate amount of raw drug powder with 10% starch paste by weight to form a soft material. Use an automatic pill-making machine for Chinese medicinal materials to make small granules with a diameter of 0.5~1 mm. Dry the prepared water pills to remove moisture and dry them at 60~80℃ for 8~12 hours to obtain the blood stasis relieving and pain-relieving pills.
4. A quality testing method for the blood-activating and pain-relieving pill according to claim 1 or claim 2, or for the blood-activating and pain-relieving pill prepared by the preparation method according to claim 3, characterized in that: The quality testing method includes: (1) General inspections: These include: disintegration degree test, moisture content test and weight difference test; (2) Microscopic identification; (3) Identification by thin-layer chromatography (TCL); and (4) High performance liquid chromatography (HPLC) content determination.
5. The quality inspection method according to claim 4, characterized in that: In (1) general inspection, the disintegration degree test adopts the disintegration time limit test method specified in the 2020 edition of the Chinese Pharmacopoeia, using a lifting disintegration apparatus, and the disintegration time of the Zhuyu Zhitong Pill is less than 8 minutes; the moisture content test adopts the drying method, and the moisture content of the Zhuyu Zhitong Pill is less than 5%; the weight difference test adopts the weight analysis test method specified in the 2020 edition of the Chinese Pharmacopoeia, and the weight difference of the Zhuyu Zhitong Pill is between 0.3% and 0.98%.
6. The quality inspection method according to claim 4, characterized in that: In (2) microscopic identification, the powder of Zhuyu Zhitong Pill is light brown. According to the 2020 edition of the Chinese Pharmacopoeia, the powder of Zhuyu Zhitong Pill is identified by microscopic identification method.
7. The quality inspection method according to claim 4, characterized in that: In (3) TCL identification, the TCL identification includes the following steps: (i) Preparation of the test solution: Take 6g of the sample of the Zhuyu Zhitong Pill, crush it, add ethyl acetate for ultrasonic extraction, centrifuge and filter, retain the filter residue, concentrate the filtrate to 2 mL by rotary evaporation, or evaporate it to dryness and then reconstitute it to 2 mL of ethyl acetate to obtain the solution. (ii) Preparation of reference herb solution: Take an appropriate amount of Angelica sinensis and Ligusticum chuanxiong reference herbs and prepare a reference herb solution using the same method as in (i); (iii) Preparation of negative control solution: Prepare double negative pills without Angelica sinensis and Ligusticum chuanxiong according to the preparation method of Zhuyu Zhitong Pills. Weigh 6g of the pills and prepare double negative control solution without Angelica sinensis and Ligusticum chuanxiong in the same way as (i). (iv) According to the thin-layer chromatography method under General Chapter 0502 of Part IV of the 2020 edition of the Chinese Pharmacopoeia, take 5-8 μL of the test solution and the reference solution and spot them separately on the same silica gel G thin-layer plate. Use n-hexane:ethyl acetate volume ratio of 4:1 as the developing solvent. After development, remove and air dry, and observe under a 365 nm ultraviolet lamp. The results show that the chromatogram of the test sample and the reference sample show the same fluorescent spots at the same Rf value.
8. The quality inspection method according to claim 7, characterized in that: In (3) TCL identification, in the Angelica sinensis-Ligusticum chuanxiong double negative system, the test sample and the control medicinal material both showed characteristic blue-green fluorescent spots at the same color development site Rf=0.
69. The negative control of Ligusticum chuanxiong and the negative control of Angelica sinensis obtained corresponding characteristic spots with Rf values of 0.66 and 0.68, respectively.
9. The quality inspection method according to claim 4, characterized in that: In (4) HPLC content determination, the HPLC content determination includes the following steps: (a) Preparation of reference solution: Take an appropriate amount of ferulic acid reference standard, accurately weigh it, place it in a brown volumetric flask, add 70% methanol to prepare a solution, and you will get the solution. (b) Preparation of test solution: Take the powder of Zhuyu Zhitong Pill that has passed through No. 3 sieve, weigh it accurately and place it in a stoppered conical flask. Add an appropriate amount of 70% methanol accurately and quantitatively, turn on the constant temperature reflux device, process for 30 min, cool to room temperature and weigh it again, add 70% methanol to the initial total mass, let it stand and separate into layers, take the supernatant and filter it through a 0.45 μm microporous membrane, collect the filtrate for testing; (c) Chromatographic conditions: Column: C18 column, mobile phase: acetonitrile-0.085% phosphoric acid solution (volume ratio 17:83), flow rate: 1.0 mL / min, detection wavelength: 316 nm, column temperature: 35℃, theoretical plate number: ferulic acid peak ≥5000.
10. The quality inspection method according to claim 9, characterized in that: In (4) HPLC content determination, ferulic acid showed good linearity in the range of 2~40 μg / mL.