A traditional Chinese medicine composition for treating chronic gastritis, its preparation method, and its detection method.

Abstract

This invention relates to the field of traditional Chinese medicine (TCM) technology, specifically to a TCM composition for strengthening the spleen and stomach, its preparation method, and its quality testing method. The TCM composition for strengthening the spleen and stomach provided by this invention comprises the following raw materials in parts by weight: Astragalus membranaceus 15-25 parts by weight, Taraxacum mongolicum 15-25 parts by weight, Angelica sinensis 6-15 parts by weight, Atractylodes macrocephala 10-20 parts by weight, Curcuma zedoaria 6-15 parts by weight, Citrus aurantium 6-15 parts by weight, Paeonia lactiflora 10-20 parts by weight, Coptis chinensis 2-8 parts by weight, Cinnamomum cassia 2-8 parts by weight, and Glycyrrhiza uralensis 6-15 parts by weight. The above-mentioned TCM composition has the effects of strengthening the spleen and stomach, clearing heat and resolving dampness, and is particularly effective in treating chronic non-atrophic gastritis complicated with Helicobacter pylori infection. This formula is significantly effective in eliminating H. pylori infection and alleviating the main symptoms of abdominal distension, abdominal pain, acid reflux, and heartburn.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a traditional Chinese medicine composition for strengthening the spleen and stomach, its preparation method, and its quality testing method. Background Technology

[0002] Chronic gastritis is one of the most common digestive system diseases, referring to various chronic inflammations of the gastric mucosa caused by different etiologies. Common clinical manifestations include nonspecific indigestion, such as upper abdominal discomfort, bloating, dull pain, and burning pain. These symptoms generally do not have a clear rhythm and may be aggravated or relieved by eating. Other symptoms may include loss of appetite, belching, acid reflux, and nausea. Erosion of the gastric mucosa can lead to upper gastrointestinal bleeding, and long-term minor bleeding can cause iron deficiency anemia.

[0003] Helicobacter pylori (HP) is the most common cause of chronic gastritis because: ① HP can be detected in gastric mucosal biopsy specimens from most (80%–95%) patients with chronic gastritis; the additional 5%–20% negative rate reflects the diversity of causes of chronic gastritis; ② The distribution of HP in the stomach is consistent with the distribution of inflammation in the stomach; ③ Eradication of HP can reduce gastric mucosal inflammation, with neutrophils generally regressing faster, while lymphocytes and plasma cells take longer to regress; ④ Chronic gastritis caused by HP infection can be replicated in volunteers and animal models; ⑤ The bacterial density of HP infection is positively correlated with the severity of chronic gastritis.

[0004] A small percentage of chronic non-atrophic gastritis can develop into chronic multifocal atrophic gastritis. In the latter, most are prone to developing gastric ulcers, and a minority may progress to gastric cancer over a long period. Comprehensive treatment, including eradication of *Helicobacter pylori* (HP) and antioxidant supplementation, can, to some extent, prevent the occurrence and development of gastric mucosal atrophy and intestinal metaplasia. Therefore, emphasizing the treatment of *Helicobacter pylori*-associated chronic non-atrophic gastritis has significant practical implications.

[0005] Currently, there are three main types of drugs used domestically and internationally to treat chronic non-atrophic gastritis with *Helicobacter pylori* (HP) infection: acid suppressants, bismuth preparations, and antibiotics. Common regimens include triple therapy and quadruple therapy. Triple therapy typically involves one bismuth preparation or proton pump inhibitor (PPI) plus two antibiotics, such as amoxicillin, metronidazole, clarithromycin, or tetracycline, for a course of 1-2 weeks. Triple therapy achieves an HP eradication rate of over 80%, but its main drawbacks include: HP resistance to metronidazole, which reduces efficacy; a high incidence of side effects; frequent medication administration leading to poor patient compliance; and high cost, making it unaffordable for most patients. Quadruple therapy, using an H2 receptor antagonist or PPI + bismuth preparation + two antibiotics, or ranitidine bismuth citrate + two antibiotics, can further improve efficacy to an eradication rate of over 90%. It has fewer side effects, is convenient to administer, and can be considered the first-line treatment when available. Its disadvantage is its high cost, which limits its widespread application. Therefore, exploring and finding more effective treatments and drugs is a hot topic and focus of research on the treatment of HP infection-related ulcers.

[0006] Both Western and Traditional Chinese Medicine (TCM) have their unique approaches to treating non-atrophic chronic gastritis. Western medicine excels at treating the root cause of the disease, with definite and targeted efficacy. TCM addresses both the symptoms and the underlying cause, providing targeted treatment with fewer side effects. For the treatment of chronic non-atrophic gastritis complicated by Helicobacter pylori infection, Western medicine consistently employs triple or quadruple therapy, but this approach has limitations, significant side effects, and a high recurrence rate.

[0007] In recent years, extensive and in-depth research on traditional Chinese medicine (TCM) and years of clinical practice by clinicians have confirmed that it not only has good clinical efficacy but also compensates for the shortcomings of Western medicine, such as poor clinical efficacy and significant side effects. Therefore, the application of TCM in the treatment of chronic non-atrophic gastritis has received increasing attention. To this end, this invention provides a TCM composition for the treatment of chronic non-atrophic gastritis. Summary of the Invention

[0008] Therefore, the technical problem to be solved by the present invention is to provide a traditional Chinese medicine composition for strengthening the spleen and stomach, a preparation method, and a quality testing method.

[0009] Therefore, the present invention provides the following technical solution:

[0010] A traditional Chinese medicine composition for strengthening the spleen and stomach, characterized in that it comprises the following raw materials in parts by weight:

[0011] Astragalus membranaceus 15-25 parts by weight, dandelion 15-25 parts by weight, angelica sinensis 6-15 parts by weight, atractylodes macrocephala 10-20 parts by weight, turmeric 6-15 parts by weight, immature bitter orange 6-15 parts by weight, red peony root 10-20 parts by weight, coptis chinensis 2-8 parts by weight, cinnamon 2-8 parts by weight, licorice root 6-15 parts by weight.

[0012] Optional, the raw materials may include the following parts by weight: Astragalus membranaceus 20 parts by weight, Taraxacum mongolicum 20 parts by weight, Angelica sinensis 10 parts by weight, Atractylodes macrocephala 15 parts by weight, Curcuma zedoaria 10 parts by weight, Citrus aurantium 10 parts by weight, Paeonia lactiflora 15 parts by weight, Coptis chinensis 5 parts by weight, Cinnamomum cassia 5 parts by weight, and Glycyrrhiza uralensis 10 parts by weight; or

[0013] Astragalus membranaceus 15 parts by weight, dandelion 15 parts by weight, angelica sinensis 6 parts by weight, atractylodes macrocephala 10 parts by weight, turmeric 6 parts by weight, immature bitter orange 6 parts by weight, red peony root 10 parts by weight, coptis chinensis 2 parts by weight, cinnamon 2 parts by weight, licorice root 6 parts by weight; or

[0014] Astragalus membranaceus 18 parts by weight, dandelion 18 parts by weight, angelica sinensis 9 parts by weight, atractylodes macrocephala 14 parts by weight, turmeric 9 parts by weight, immature bitter orange 9 parts by weight, red peony root 14 parts by weight, coptis chinensis 4 parts by weight, cinnamon 4 parts by weight, licorice root 9 parts by weight; or

[0015] Astragalus membranaceus 17 parts by weight, dandelion 17 parts by weight, angelica sinensis 8 parts by weight, atractylodes macrocephala 12 parts by weight, turmeric 8 parts by weight, immature bitter orange 8 parts by weight, red peony root 12 parts by weight, coptis chinensis 4 parts by weight, cinnamon 4 parts by weight, and licorice root 8 parts by weight.

[0016] A method for preparing the aforementioned traditional Chinese medicine composition includes the following steps:

[0017] (1) Weigh out the volatile oils of Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium and Cinnamomum cassia according to the selected weight proportions, add water and distill to extract the volatile oils, collect the volatile oils, encapsulate them, filter the distilled aqueous solution and set aside;

[0018] (2) Weigh out dandelion, red peony root and licorice according to the selected weight proportions, add water to extract, filter, and use the filtrate for later use;

[0019] (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the selected weight proportions, extract with ethanol solution, filter, and keep the filtrate for later use;

[0020] (4) Mix the filtrates obtained in steps (2) and (3) and dry them. Add volatile oil inclusion complex and then add or not add conventional excipients to make a clinically acceptable dosage form.

[0021] Optionally, in the inclusion step of step (1), the ratio of volatile oil, β-cyclodextrin and water is 1ml: 5-20g: 40-100ml; the inclusion time is 5-30 minutes; and the inclusion mixture is refrigerated for 12-48 hours after inclusion.

[0022] Optionally, in step (1) of water distillation extraction, 8-14 times the amount of water is added and steam distillation is carried out for 4-8 hours;

[0023] And / or, in the water extraction step (2), add water and decoct 1-3 times, 1-3 hours each time, adding 6-14 times the amount of water each time;

[0024] And / or, in step (3) ethanol extraction, the extract is heated and refluxed with a 50 v / v%-80 v / v% ethanol solution for 1-3 times, each time for 1-2 hours, with 6-12 times the volume of the 50 v / v%-80 v / v% ethanol solution added each time. The extract is filtered and combined.

[0025] And / or, the dosage form includes tablets, granules, capsules, oral liquids, or injections.

[0026] A method for quality testing of the spleen-strengthening and stomach-regulating traditional Chinese medicine composition or the spleen-strengthening and stomach-regulating traditional Chinese medicine composition prepared by the aforementioned preparation method, comprising the following content determination methods:

[0027] A. Determination of astragaloside A content:

[0028] Chromatographic conditions: Determined by high performance liquid chromatography (HPLC), using octadecylsilane-bonded silica gel as the packing material; acetonitrile as mobile phase A and water as mobile phase B, with a volume ratio of mobile phase A:mobile phase B of 20-40%:80-60%; drift tube temperature of the evaporative light scattering detector 35-100℃; gas pressure 300-400 kPa; theoretical plate number calculated based on astragaloside A should not be less than 5000.

[0029] Determination method: Pipe the reference solution and the test solution separately into the liquid chromatograph, measure and calculate.

[0030] Optionally, the determination of the content of A and astragaloside A further includes:

[0031] Preparation of the test solution: Take the Chinese herbal composition, add methanol solution, heat to reflux, cool, filter, evaporate the filtrate to dryness, dissolve the residue in water, extract with water-saturated n-butanol, combine the n-butanol solutions, wash, discard the washing liquid, evaporate to dryness, dissolve the residue in methanol and make up to volume to obtain the test solution.

[0032] Preparation of reference solution: Take astragaloside A reference standard and add organic solvent to prepare reference solution;

[0033] Optionally, the concentration of the methanol solution is 70-100 v / v%.

[0034] Optionally, the traditional Chinese medicine composition has a mass of 5g, and the amount of methanol solution used is 20-150ml;

[0035] Optionally, the heating reflux time is 0.1-1.0 h;

[0036] Optionally, extract with water-saturated n-butanol 1-5 times, 10-50ml each time;

[0037] Optionally, the residue can be dissolved in methanol and brought to a final volume of 2-50 ml.

[0038] Optionally, it may also include at least one of the following content determination methods or identification methods:

[0039] B. Determination of the content of paeoniflorin and naringin:

[0040] Chromatographic conditions: The determination was performed using high performance liquid chromatography (HPLC), with octadecylsilane-bonded silica gel as the stationary phase; acetonitrile as mobile phase A and 0.05-0.4 v / v% phosphoric acid solution as mobile phase B, with gradient elution. The volume ratio of mobile phase A to mobile phase B was 5-80%:95-20%; the detection wavelength was 220-290 nm; the theoretical plate number, calculated based on naringin, should be no less than 5000.

[0041] Assay: Accurately pipette the reference solution and the test solution separately and inject them into the liquid chromatograph; measure and calculate; or

[0042] C. Identification of Angelica sinensis:

[0043] Preparation of the test solution: Take 0.5-4g of the traditional Chinese medicine composition, add 10-40ml of ether, sonicate for 5-30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1-5ml of ethanol to obtain the test solution;

[0044] Preparation of reference medicinal material solution: Take 0.1-2g of Angelica sinensis reference medicinal material and prepare the reference medicinal material solution according to the preparation method of the test solution;

[0045] Identification: Perform thin-layer chromatography (TLC). Apply 2-10 μl each of the test solution and the reference medicinal material solution to the same silica gel G TLC plate. Develop the plate using a mixture of n-hexane and ethyl acetate in a volume ratio of 2-5:1-3. Remove the plate, air-dry, and examine. In the chromatogram of the test sample, fluorescent spots of the same color should appear at the corresponding positions as in the chromatogram of the reference medicinal material.

[0046] D. Identification of Atractylodes macrocephala:

[0047] Preparation of the test solution: Take 0.5-4g of the traditional Chinese medicine composition, add 5-30ml of n-hexane, sonicate for 10-60 minutes, cool, filter, and take the filtrate to obtain the test solution;

[0048] Preparation of the reference herb solution: Take 0.5-2g of Atractylodes macrocephala reference herb and prepare the reference herb solution according to the preparation method of the test sample;

[0049] Identification: Perform thin-layer chromatography (TLC). Apply 1-10 μl each of the test solution and the reference medicinal material solution to the same silica gel G TLC plate. Develop the plate using a mixture of petroleum ether and ethyl acetate at a volume ratio of 20-100:1-5 (60-90℃). Remove the plate, air dry, spray with 1-10% w / v vanillin-sulfuric acid solution, and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material; or...

[0050] E. Identification of Citrus aurantium:

[0051] Preparation of the test solution: Take 0.5-4g of the Chinese herbal composition, add 5-30ml of 40-100v / v% methanol, sonicate for 10-60 minutes, cool, filter, evaporate the filtrate to dryness, add 1-5ml of methanol to dissolve the residue to obtain the test solution.

[0052] Preparation of reference solutions: Take naringin reference standard and neohesperidin reference standard, add 40-100 v / v% methanol to prepare solutions containing 0.2-2 mg per 1 ml, respectively, as reference solutions;

[0053] Identification: Perform thin-layer chromatography. Apply 1-10 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Use the lower layer of a mixture of chloroform, methanol, and water in a volume ratio of 10-15:2-10:1-4 as the developing solvent. Develop to a distance of 8-20 cm. Remove, air dry, and spray with 1-5% w / v aluminum chloride ethanol solution. Heat at 100-110°C (105°C) for approximately 3-6 minutes. Examine under ultraviolet light. The test sample chromatogram should show fluorescent spots of the same color at the corresponding positions as the reference sample chromatogram; or...

[0054] F. Identification of Paeonia lactiflora:

[0055] Preparation of the test solution: Take 0.5-4g of the Chinese herbal composition, add 5-30ml of 40-100% ethanol, shake for 5-30 minutes, filter, evaporate the filtrate to dryness, and dissolve the residue in 1-5ml of 40-100% ethanol to obtain the test solution.

[0056] Preparation of reference solution: Take paeoniflorin reference standard and add 40-100 v / v% methanol to prepare a solution containing 0.2-2 mg per 1 ml, which is used as the reference solution;

[0057] Identification: Perform thin-layer chromatography. Apply 1-10 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Use a mixture of chloroform-ethyl acetate-methanol-formic acid (v / v) at a ratio of 20-50:2-10:5-15:0.1-1 as the developing solvent. Develop, remove, and air-dry. Spray with 5% vanillin-sulfuric acid-ethanol solution and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference sample; or...

[0058] G. Identification of Coptis chinensis:

[0059] Preparation of the test solution: Take 0.5-4g of the traditional Chinese medicine composition, add 5-30ml of 40-100v / v% methanol, sonicate for 5-30 minutes, filter, and take the filtrate to obtain the test solution;

[0060] Preparation of reference solution: Take berberine hydrochloride reference standard and add 40-100 v / v% methanol to prepare a solution containing 0.2-2 mg per 1 ml, which is used as the reference solution;

[0061] Identification: Perform thin-layer chromatography. Apply 1-10 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Use a mixture of cyclohexane, ethyl acetate, isopropanol, methanol, water, and triethylamine in a volume ratio of 1-5:1-5:0.5-3:0.5-3:0.5-3:0.5-3 as the developing solvent. Place the plate in a developing tank pre-saturated with concentrated ammonia solution for 20 minutes. Develop, remove, air dry, and examine under ultraviolet light. The test sample chromatogram should show fluorescent spots of the same color at the corresponding positions as the reference chromatogram.

[0062] Optionally, in the determination of the contents of B, paeoniflorin, and naringin:

[0063] The gradient elution program is as follows: 0–9 min, 15% A; 9–30 min, 15% A–22% A; 30–33 min, 22% A–50% A; 33–35 min, 50% A;

[0064] And / or, preparation of the test solution: Take 0.5-3g of the traditional Chinese medicine composition, add 10-100mL of 40-100% methanol, seal tightly, weigh, reflux in a water bath for 1-3 hours, cool, weigh again, replenish the lost weight with 40-100% methanol, shake well, filter, and take the filtrate to obtain the test solution;

[0065] And / or, preparation of reference solutions: Take appropriate amounts of paeoniflorin and naringin reference standards, accurately weigh them, and add 50-100% methanol solution to prepare paeoniflorin and naringin reference solutions.

[0066] And / or, the injection volume is 2-20 μL.

[0067] The spleen-strengthening and stomach-regulating traditional Chinese medicine composition or the spleen-strengthening and stomach-regulating traditional Chinese medicine composition prepared by the aforementioned preparation method has any of the following uses:

[0068] (1) Use in the preparation of medicines that have the functions of strengthening the spleen and stomach, clearing heat and removing dampness;

[0069] (2) Use in the preparation of drugs for treating chronic non-atrophic gastritis complicated with Helicobacter pylori infection;

[0070] (3) Use in the preparation of drugs for treating Helicobacter pylori infection;

[0071] (4) Use in the preparation of drugs for treating bloating, stomach pain, acid reflux and heartburn;

[0072] (5) Use in the preparation of drugs for treating acute inflammation.

[0073] This invention relates to a traditional Chinese medicine composition developed based on the clinical experience formula of Professor Zhou Ping'an, a renowned TCM expert at Beijing University of Traditional Chinese Medicine, for treating chronic non-atrophic gastritis—Qi Ying Bao Wei Fang (Astragalus and Evodia Stomach-Protecting Formula). Professor Zhou believes that chronic non-atrophic gastritis is often related to external pathogenic factors (including H. pylori infection), internal emotional factors, and dietary and physical strain, with spleen and stomach damage and impaired spleen function being the root cause. In the early stages of stomach disease, it is often a case of excess, with the disease location in the stomach; in chronic cases, it is often a case of deficiency, with the disease location in the spleen. A mixture of excess and deficiency is common, and simultaneous spleen and stomach involvement is also frequently seen. The disease is caused by improper diet, excessive fatigue, excessive blood loss, or prolonged illness damaging the spleen and stomach, or by inherent weakness of the body, resulting in impaired spleen and stomach function, weakened digestive and atrophic function, and obstructed qi flow. Weakness of the spleen and stomach leads to impaired digestion and absorption, resulting in stomach dysfunction and dampness obstructing the middle burner. This stagnation of Qi causes abdominal distension and pain. "Excess Qi becomes fire," and prolonged stagnation of stomach Qi transforms into heat, causing burning pain in the stomach. Heat scorches body fluids, leading to dry mouth and bitter taste. Upward reversal of stomach Qi results in belching, acid reflux, and poor appetite. The spleen is the source of Qi and blood; deficiency leads to weakness of Qi and blood, fatigue, and lethargy. Furthermore, due to prolonged illness affecting the collaterals and blood stasis, the symptoms often include damp-heat, Qi stagnation, and blood stasis. Therefore, the treatment principle is to strengthen the spleen and stomach, clear heat and resolve dampness, and promote Qi and blood circulation. Through long-term clinical practice, a formula combining raw Astragalus membranaceus, dandelion, Angelica sinensis, raw Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium, Paeonia lactiflora, Coptis chinensis, Cinnamomum cassia, and raw Glycyrrhiza uralensis to protect the stomach has been developed.

[0074] The technical solution of this invention has the following advantages:

[0075] 1. This invention provides a traditional Chinese medicine composition for strengthening the spleen and stomach, comprising the following raw materials in parts by weight: Astragalus membranaceus 15-25 parts by weight, Taraxacum mongolicum 15-25 parts by weight, Angelica sinensis 6-15 parts by weight, Atractylodes macrocephala 10-20 parts by weight, Curcuma zedoaria 6-15 parts by weight, Citrus aurantium 6-15 parts by weight, Paeonia lactiflora 10-20 parts by weight, Coptis chinensis 2-8 parts by weight, Cinnamomum cassia 2-8 parts by weight, and Glycyrrhiza uralensis 6-15 parts by weight; Astragalus membranaceus and Taraxacum mongolicum are the principal herbs, invigorating qi and strengthening the spleen, clearing heat and promoting diuresis; Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium, Paeonia lactiflora, and Angelica sinensis are the assistant herbs, invigorating the spleen and regulating qi, promoting blood circulation and removing blood stasis, regulating the stomach and relieving bloating and lowering qi, relieving spasms and pain; Coptis chinensis and Cinnamomum cassia are the adjuvant herbs, pungent and bitter, regulating cold and heat; Glycyrrhiza uralensis is the guiding herb, tonifying qi and detoxifying, relieving spasms and pain, and harmonizing the other herbs. Astragalus membranaceus in this formula has the function of invigorating qi and supporting the body's resistance. Dandelion clears heat and detoxifies, disperses nodules and eliminates carbuncles. When combined with Astragalus membranaceus, they complement each other, one tonifying and the other clearing, clearing heat and promoting diuresis, and invigorating the spleen and qi. Atractylodes macrocephala works synergistically with Astragalus membranaceus to enhance the effects of invigorating qi and strengthening the spleen, as well as drying dampness and neutralizing acid. Paeonia lactiflora works synergistically with Angelica sinensis to nourish and invigorate blood, promoting the recovery of gastric mucosal function. Curcuma zedoaria combined with Astragalus membranaceus ensures that Astragalus membranaceus tonifies without causing stagnation, ensuring sufficient qi in the middle jiao, and making the effects of Curcuma zedoaria in regulating qi and relieving pain even stronger. Citrus aurantium combined with Astragalus membranaceus invigorates qi and strengthens the spleen, regulates qi and promotes qi circulation, and harmonizes the stomach. Coptis chinensis clears stomach heat, drains damp fire, and strengthens the stomach and intestines, while assisting dandelion in clearing heat and detoxifying. Cinnamomum cassia invigorates spleen yang, warms the middle jiao and relieves pain, warms the meridians and promotes blood circulation, and assists Astragalus membranaceus and Atractylodes macrocephala in promoting tissue regeneration and anti-inflammation. Therefore, the above-mentioned Chinese herbal medicine composition has the effects of strengthening the spleen and stomach, clearing heat and eliminating dampness. It has a significant therapeutic effect, especially in treating chronic non-atrophic gastritis complicated with Helicobacter pylori infection. The formula is effective in eliminating HP infection and relieving patients' main symptoms such as abdominal distension, abdominal pain, acid reflux and heartburn.

[0076] 2. The preparation method of the spleen-strengthening and stomach-nourishing traditional Chinese medicine composition provided by the present invention includes the following steps: The preparation method of the traditional Chinese medicine composition includes the following steps: (1) According to the selected weight parts, Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium and Cinnamomum cassia are added and water is distilled to extract volatile oil, the volatile oil is collected, packaged, and the aqueous solution after distillation is filtered and set aside; (2) According to the selected weight parts, Taraxacum mongolicum, Paeonia lactiflora and Glycyrrhiza uralensis are added and water is extracted, filtered, and the filtrate is set aside; (3) According to the selected weight parts, Astragalus membranaceus and Coptis chinensis are added and ethanol solution is extracted, filtered, and the filtrate is set aside; (4) The filtrates obtained in steps (2) and (3) are mixed and dried to make a dry powder, and then conventional excipients are added or not added to make a clinically acceptable dosage form; The above-mentioned preparation method of the spleen-strengthening and stomach-nourishing traditional Chinese medicine composition is simple and easy to operate. By adjusting the raw materials and selecting a suitable extraction method, the content of effective ingredients in the prepared traditional Chinese medicine composition is increased, thereby improving the therapeutic effect on chronic non-atrophic gastritis complicated with Helicobacter pylori infection.

[0077] 3. This invention provides a method for quality testing of a traditional Chinese medicine composition for strengthening the spleen and stomach, including the following content determination method: Astragaloside A content determination: Chromatographic conditions: determined by high performance liquid chromatography, using octadecylsilane-bonded silica gel as the packing material; acetonitrile as mobile phase A, water as mobile phase B, with a volume ratio of mobile phase A:mobile phase B of 20-40%:80-60%; evaporative light scattering detector drift tube temperature 35-100℃; gas pressure 300-400 kPa; theoretical plate number calculated based on astragaloside A should not be less than 5000; Determination method: inject the reference solution and the test solution into the liquid chromatograph, respectively, and determine and calculate; by determining the content of the effective component astragaloside A in the traditional Chinese medicine composition, effective monitoring of product quality is achieved. This method effectively separates astragaloside A from other impurity peaks in the test solution, overcoming the limitations of traditional Chinese medicine (TCM) composition quality detection due to the numerous flavors, complex components, and mutual interference of TCM ingredients. It obtains the chromatographic peak of astragaloside A, the active ingredient in the TCM composition. The determined HPLC content determination method is not only highly specific, but its linearity, precision, and recovery rate also meet the requirements of pharmaceutical analysis. This achieves the goal of controlling the content of TCM composition or preparation by determining the content of the active ingredient astragaloside A. It enables comprehensive and clear quality detection of the TCM composition, further realizing quality control. Furthermore, this quality detection method has the advantages of being simple, rapid, stable, reliable, highly precise, and easy to master.

[0078] 4. This invention provides a method for quality testing of a traditional Chinese medicine composition for strengthening the spleen and stomach, comprising the preparation of a test solution: taking the traditional Chinese medicine composition, adding methanol solution, heating under reflux, cooling, filtering, evaporating the filtrate to dryness, dissolving the residue in water, extracting with water-saturated n-butanol, combining the n-butanol solutions, washing, discarding the washing liquid, evaporating to dryness, dissolving the residue in methanol and making up to volume to obtain the test solution; using the above method to prepare the test solution results in a high extraction rate of astragaloside A, which can further improve the accuracy of quality testing.

[0079] 5. This invention provides a quality detection method for a traditional Chinese medicine composition for strengthening the spleen and stomach. By simultaneously determining the contents of paeoniflorin and naringin, the comprehensiveness and reliability of quality monitoring of the traditional Chinese medicine composition are improved. By screening and obtaining a suitable extraction method to extract the traditional Chinese medicine composition to prepare a test solution, and selecting a suitable mobile phase, the finally determined HPLC content determination method not only has strong specificity, but also meets the requirements of drug analysis in terms of linearity, precision and recovery rate. It can quickly and accurately determine the quality of the product and effectively monitor the quality performance of the product.

[0080] 6. This invention provides a quality testing method for a traditional Chinese medicine composition for strengthening the spleen and stomach. The method uses TLC to identify Angelica sinensis, Atractylodes macrocephala, Citrus aurantium, Paeonia lactiflora, and Coptis chinensis, making the quality testing of the traditional Chinese medicine composition for strengthening the spleen and stomach more comprehensive and clear. This method more effectively characterizes the effective active components and their quality, facilitating comprehensive monitoring of the active ingredients and further ensuring the stability, consistency, and controllability of the quality of the traditional Chinese medicine composition for strengthening the spleen and stomach, as well as ensuring the safety and efficacy of the traditional Chinese medicine composition. Attached Figure Description

[0081] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0082] Figure 1 This is a thin-layer chromatogram of the traditional Chinese medicine composition described in Example 10 of the present invention; from left to right in the figure are: test sample 1, Angelica sinensis negative sample, test sample 2, test sample 3, and Angelica sinensis reference material;

[0083] Figure 2 This is a thin-layer chromatogram of the traditional Chinese medicine composition described in Example 11 of the present invention; from left to right in the figure are: Atractylodes macrocephala reference material, test sample 1, test sample 2, test sample 3, and Atractylodes macrocephala negative sample;

[0084] Figure 3 The image shows a thin-layer chromatogram of the traditional Chinese medicine composition described in Example 12 of this invention; from left to right in the image are: neohesperidin reference standard, naringin reference standard, test sample 1, test sample 2, test sample 3, and bitter orange negative sample;

[0085] Figure 4 The image shows a thin-layer chromatogram of the traditional Chinese medicine composition described in Example 13 of this invention; from left to right in the image are: paeoniflorin reference standard, test sample 1, test sample 2, test sample 3, and red peony negative sample;

[0086] Figure 5 This is a thin-layer chromatogram of the traditional Chinese medicine composition described in Example 14 of the present invention; from left to right in the figure are: berberine hydrochloride reference standard, test sample 1, test sample 2, test sample 3, and Coptis chinensis negative sample;

[0087] Figure 6 This is the high-performance liquid chromatogram from Experimental Example 6 of the present invention; wherein A is the chromatogram of the reference solution; B is the chromatogram of the test solution; and C is the chromatogram of the negative control solution.

[0088] Figure 7This is the high-performance liquid chromatogram of paeoniflorin in Experimental Example 7 of the present invention; wherein A is the chromatogram of the reference solution; B is the chromatogram of the test solution; and C is the chromatogram of the negative control solution.

[0089] Figure 8 This is the high-performance liquid chromatogram of naringin in Experimental Example 7 of the present invention; wherein A is the chromatogram of the reference solution; B is the chromatogram of the test solution; and C is the chromatogram of the negative control solution.

[0090] Figure 9 This is the standard curve of astragaloside A obtained in the linearity investigation of Experiment Example 6 of this invention;

[0091] Figure 10 This is the linear curve of paeoniflorin obtained in the investigation of linear relationship in Experiment Example 7 of this invention;

[0092] Figure 11 This is the linear curve of naringin obtained in the investigation of linear relationship in Experiment Example 7 of this invention;

[0093] Figure 12 These are pathological photomicrographs (HE×200) of the gastric tissue from the blank group in Experiment Example 1 of this invention:

[0094] Figure 13 These are pathological photomicrographs (HE×200) of the gastric tissue from the model group in Experiment Example 1 of this invention:

[0095] Figure 14 These are pathological photomicrographs (HE×200) of the gastric tissue from the aluminum magnesium carbonate tablet group in Experimental Example 1 of this invention:

[0096] Figure 15 These are pathological micrographs (HE×200) of the gastric tissue from the stomach-strengthening and anti-inflammatory granule group in Experimental Example 1 of this invention:

[0097] Figure 16 These are pathological micrographs (HE×200) of the gastric tissue from the high-dose group of the traditional Chinese medicine composition in Experimental Example 1 of this invention:

[0098] Figure 17 These are pathological micrographs (HE×200) of the gastric tissue in the medium-dose group of the traditional Chinese medicine composition in Experimental Example 1 of this invention:

[0099] Figure 18 These are pathological micrographs (HE×200) of the gastric tissue from the low-dose group of the traditional Chinese medicine composition in Experimental Example 1 of this invention. Detailed Implementation

[0100] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0101] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0102] Example

[0103] The formulations of the spleen-strengthening and stomach-nourishing traditional Chinese medicine compositions of Examples 1-7 are shown in the table below:

[0104] Table 1. Formulation of Traditional Chinese Medicine Compositions

[0105]

[0106] Preparation method of the traditional Chinese medicine composition in Example 1:

[0107] (1) Weigh out Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium and Cinnamomum cassia according to the formula, add 14 times the amount of water, steam distill for 6 hours to extract volatile oil, collect the volatile oil, filter the distilled aqueous solution and set aside; add β-cyclodextrin and water to the volatile oil according to the ratio of volatile oil, β-cyclodextrin and water as 1ml:10g:50ml, stir, incorporate for 10 minutes, refrigerate for 24 hours, filter, dry at 40℃ to obtain volatile oil inclusion complex and set aside;

[0108] (2) Weigh out dandelion, red peony root and licorice according to the formula, add water and decoct twice, one hour each time, adding 10 times the amount of water each time. Filter the decoction and keep the filtrate for later use.

[0109] (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the formula and extract them three times by heating and refluxing with 60 v / v% ethanol solution for 2 hours each time. Add 10 times the amount of 60 v / v% ethanol solution each time. Filter the extract, combine the filtrates, and recover the ethanol.

[0110] (4) Combine the filtrates from steps (3) and (4), concentrate them to a relative density of 1.30-1.35 (50℃), dry them, pulverize them into fine powder, sieve them, add volatile oil inclusion complex, then add appropriate amounts of aspartame and lactose, mix well, make granules, dry them, and you will get the product.

[0111] Preparation method of the traditional Chinese medicine composition in Example 2:

[0112] (1) Weigh out the above ten ingredients according to the formula, take Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium, and Cinnamomum cassia, add 12 times the amount of water, steam distill for 5 hours to extract volatile oil, collect the volatile oil, filter the distilled aqueous solution and set aside; add β-cyclodextrin and water to the volatile oil at a ratio of 1ml:5g:40ml, stir, incorporate for 15 minutes, refrigerate for 30 hours, filter, dry at 45℃ to obtain the volatile oil inclusion complex and set aside;

[0113] (2) Weigh out dandelion, red peony root and licorice root according to the formula, add water and decoct twice, 1.5 hours each time, add 6 times the amount of water each time, filter the decoction and keep the filtrate for later use.

[0114] (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the formula and extract them three times by heating and refluxing with 70 v / v% ethanol solution for 1 hour each time. Add 12 times the amount of 70 v / v% ethanol solution each time. Filter the extract, combine them, and recover the ethanol.

[0115] (4) Combine the filtrates from steps (3) and (4), concentrate them to a relative density of 1.20-1.25 (50℃), dry them, pulverize them into fine powder, sieve them, add volatile oil inclusion complex, then add appropriate amounts of aspartame and dextrin, mix well, make granules, dry them, and you will get the product.

[0116] Preparation method of the traditional Chinese medicine composition in Example 3:

[0117] (1) Weigh out Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium and Cinnamomum cassia according to the formula, add 10 times the amount of water, steam distill for 4 hours to extract volatile oil, collect the volatile oil, filter the distilled aqueous solution and set aside; add β-cyclodextrin and water to the volatile oil according to the ratio of volatile oil, β-cyclodextrin and water as 1ml:6g:60ml, stir, incorporate for 25 minutes, refrigerate for 40 hours, filter, dry at 50℃ to obtain volatile oil inclusion complex and set aside;

[0118] (2) Weigh out dandelion, red peony root and licorice root according to the formula, add water and decoct once for 2 hours, add 14 times the amount of water, filter the decoction and keep the filtrate for later use.

[0119] (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the formula and extract them twice by heating and refluxing with 80 v / v% ethanol solution for 1.5 hours each time. Add 6 times the amount of 80 v / v% ethanol solution each time. Filter the extract, combine the extracts, and recover the ethanol.

[0120] (4) Combine the filtrates from steps (3) and (4), concentrate them to a relative density of 1.25-1.30 (50℃), dry them, pulverize them into fine powder, sieve them, add volatile oil inclusion complex, then add appropriate amounts of aspartame and dextrin, mix well, make granules, dry them, and you will get the product.

[0121] Preparation method of the traditional Chinese medicine composition in Example 4:

[0122] (1) Weigh out Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium and Cinnamomum cassia according to the formula, add 8 times the amount of water, steam distill for 6 hours to extract volatile oil, collect the volatile oil, filter the distilled aqueous solution and set aside; add β-cyclodextrin and water to the volatile oil according to the ratio of volatile oil, β-cyclodextrin and water as 1ml:8g:50ml, stir, incorporate for 30 minutes, refrigerate for 48 hours, filter, dry at 40℃ to obtain volatile oil inclusion complex and set aside;

[0123] (2) Weigh out dandelion, red peony root and licorice according to the formula, add water and decoct twice, one hour each time, adding 8 times the amount of water each time. Filter the decoction and keep the filtrate for later use.

[0124] (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the formula and extract them once by heating and refluxing with 50 v / v% ethanol solution for 2 hours each time. Add 8 times the amount of 50 v / v% ethanol solution each time. Filter the extract, combine them, and recover the ethanol.

[0125] (4) Combine the filtrates from steps (3) and (4), concentrate them to a relative density of 1.30-1.35 (50℃), dry them, pulverize them into fine powder, sieve them, add volatile oil inclusion complex, then add appropriate amounts of aspartame and dextrin, mix well, make granules, and fill them into capsules to obtain the final product.

[0126] Preparation method of the traditional Chinese medicine composition in Example 5:

[0127] (1) Weigh out the above ten ingredients according to the formula. Take Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium, and Cinnamomum cassia. Add 10 times the amount of water and steam distill for 6 hours to extract the volatile oil. Collect the volatile oil and filter the distilled aqueous solution for later use. Add β-cyclodextrin and water to the volatile oil at a ratio of 1 ml: 6 g: 50 ml. Stir and encapsulate for 20 minutes. Refrigerate for 12 hours. Filter and dry at 40°C to obtain the volatile oil inclusion complex for later use.

[0128] (2) Weigh out dandelion, red peony root and licorice root according to the formula, add water and decoct 3 times, 1.5 hours each time, add 12 times the amount of water each time, filter the decoction and keep the filtrate for later use.

[0129] (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the formula and extract them twice by heating and refluxing with 60 v / v% ethanol solution for 1 hour each time. Add 10 times the amount of 60 v / v% ethanol solution each time. Filter the extract, combine them, and recover the ethanol.

[0130] (4) Combine the filtrates from steps (3) and (4), concentrate them to a relative density of 1.20-1.25 (50℃), dry them, pulverize them into fine powder, sieve them, add volatile oil inclusion complex, then add appropriate amounts of aspartame and dextrin, mix well, and compress them into tablets to obtain the product.

[0131] Preparation method of the traditional Chinese medicine composition in Example 6:

[0132] (1) Weigh out Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium and Cinnamomum cassia according to the formula, add 12 times the amount of water, steam distill for 5 hours to extract volatile oil, collect the volatile oil, filter the distilled aqueous solution and set aside; add β-cyclodextrin and water to the volatile oil according to the ratio of volatile oil, β-cyclodextrin and water as 1ml:6g:60ml, stir, incorporate for 5 minutes, refrigerate for 30 hours, filter, dry at 40℃ to obtain volatile oil inclusion complex and set aside;

[0133] (2) Weigh out dandelion, red peony root and licorice according to the formula, add water and decoct twice, each time for 2 hours, each time adding 12 times the amount of water. Filter the decoction and keep the filtrate for later use.

[0134] (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the formula and extract them twice by heating and refluxing with 60 v / v% ethanol solution for 1.5 hours each time. Add 10 times the amount of 60 v / v% ethanol solution each time. Filter the extract, combine the extracts, and recover the ethanol.

[0135] (4) Combine the filtrates from steps (3) and (4), concentrate them to a relative density of 1.20-1.25 (50℃), dry them, pulverize them into fine powder, sieve them, add the volatile oil inclusion complex, mix them well, and obtain the traditional Chinese medicine composition.

[0136] Preparation method of the traditional Chinese medicine composition in Example 7:

[0137] (1) Weigh out Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium and Cinnamomum cassia according to the formula, add 9 times the amount of water, steam distill for 8 hours to extract volatile oil, collect the volatile oil, filter the distilled aqueous solution and set aside; add β-cyclodextrin and water to the volatile oil according to the ratio of volatile oil, β-cyclodextrin and water as 1ml:20g:100ml, stir, incorporate for 15 minutes, refrigerate for 40 hours, filter, dry at 40℃ to obtain volatile oil inclusion complex and set aside;

[0138] (2) Weigh out dandelion, red peony root and licorice according to the formula, add water and decoct twice, one hour each time, adding 7 times the amount of water each time. Filter the decoction and keep the filtrate for later use.

[0139] (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the formula and extract them three times by heating and refluxing with 70 v / v% ethanol solution for 1.5 hours each time. Add 12 times the amount of 70 v / v% ethanol solution each time. Filter the extract, combine the extracts, and recover the ethanol.

[0140] (4) Combine the filtrates from steps (3) and (4), concentrate them to a relative density of 1.30 (50℃), dry them, pulverize them into fine powder, sieve them, add the volatile oil inclusion complex, mix them well, and obtain the traditional Chinese medicine composition.

[0141] Example 8

[0142] This embodiment provides a quality testing method for a traditional Chinese medicine composition for strengthening the spleen and stomach, comprising the following steps: the traditional Chinese medicine composition for strengthening the spleen and stomach is prepared in Example 1;

[0143] A. Determination of Astragaloside A content

[0144] Preparation of the test solution: Weigh approximately 5g of the test sample accurately and place it in a stoppered conical flask. Add 100ml of 70v / v% methanol solution, heat under reflux for 1 hour, filter, evaporate the filtrate to dryness, add 10ml of water to the residue, heat gently to dissolve, and extract with water-saturated n-butanol by shaking 4 times, 40ml each time. Combine the n-butanol extracts and wash thoroughly twice with ammonia solution, 40ml each time. Discard the ammonia solution, evaporate the n-butanol extract to dryness, dissolve the residue in methanol, transfer it to a 5ml volumetric flask, dilute with methanol to the mark, and shake well to obtain the test solution.

[0145] Preparation of reference solution: Take an appropriate amount of astragaloside A reference standard, accurately weigh it, and add methanol solution to prepare a solution containing 0.5 mg per ml.

[0146] Chromatographic conditions: Octadecylsilane-bonded silica gel column (Agilent ZORBAX SB-C18 column (5 μm, 4.6 × 150 mm)); acetonitrile-water (32:68 v / v); evaporative light scattering detector; drift tube temperature 80 °C; gas pressure 350 kPa. The theoretical plate number, calculated based on the astragaloside A peak, should be no less than 5000.

[0147] Assay: Accurately pipette 10 μl and 20 μl of the reference solution and 20 μl of the test solution into the liquid chromatograph, respectively, and determine the result using the logarithmic equation of the external standard two-point method.

[0148] Each sachet of this product contains astragalus root extract as astragaloside A (C). 41 H 68 O 14 The total amount should not be less than 0.5mg.

[0149] B. Determination of the content of paeoniflorin and naringin

[0150] Preparation of the test solution: Take the granules prepared above, weigh them accurately, add 50 mL of 60 v / v% methanol accurately, stopper tightly, weigh them, reflux in a water bath for 2 hours, cool, weigh them again, make up the lost weight with 60 v / v% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0151] Preparation of reference solution: Take appropriate amounts of paeoniflorin and naringin reference standards, accurately weigh them, and add 60 v / v% methanol solution to prepare a solution containing 70 μg of paeoniflorin and 80 μg of naringin per 1 mL.

[0152] Chromatographic conditions: The determination was performed using high-performance liquid chromatography (HPLC), with octadecylsilane-bonded silica gel as the stationary phase; acetonitrile as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, with gradient elution: 0–9 min, 15 v / v% A; 9–30 min, 15 v / v% A → 22 v / v% A; 30–33 min, 22 v / v% A → 50 v / v% A; 33–35 min, 50 v / v% A; detection wavelengths were 230 nm and 283 nm; the theoretical plate number, calculated based on naringin, should not be less than 5000.

[0153] The assay involves precisely injecting 10 μl each of the paeoniflorin reference solution, naringin reference solution, and test solution into a liquid chromatograph and measuring the results.

[0154] Each sachet of this product contains naringin (C) from citrus aurantium. 27 H 32 O 14 The total dose shall not be less than 23.5 mg.

[0155] Each packet of this product contains paeoniflorin (C) from paeoniflorin. 23 H 28 O 11 The total dose shall not be less than 18.1 mg.

[0156] C. Identification of Angelica sinensis:

[0157] Preparation of the test solution: Take 0.5g of the particles, add 10ml of ether, sonicate (power 250W, frequency 40kHz) for 5 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1ml of ethanol to obtain the test solution.

[0158] Preparation of reference drug solution: Take 0.1g of Angelica sinensis reference drug and prepare the reference drug solution using the same method;

[0159] Angelica sinensis negative sample solution: prepared according to the preparation method of the test solution, the only difference being that the prescription does not contain Angelica sinensis;

[0160] Identification: Perform thin-layer chromatography. Apply 10 μl of each of the above solutions separately to the same silica gel G thin-layer plate. Develop with n-hexane-ethyl acetate (2:3, v / v) as the developing solvent. Remove the plate, air dry, and examine under ultraviolet light (365 nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material.

[0161] D. Identification of Atractylodes macrocephala:

[0162] Preparation of the test solution: Take 4g of the capsule, add 30ml of n-hexane, sonicate for 60 minutes, cool, filter, and take the filtrate to obtain the test solution;

[0163] Preparation of the reference herb solution: Take 2g of Atractylodes macrocephala reference herb and prepare the reference herb solution according to the preparation method of the test sample;

[0164] Atractylodes macrocephala negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the prescription does not contain Atractylodes macrocephala;

[0165] Identification: Perform thin-layer chromatography. Apply 5 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Develop the plate using a 50:1 (v / v) petroleum ether (60-90℃)-ethyl acetate solution as the developing solvent. Remove the plate, air dry, spray with 5% vanillin-sulfuric acid solution, and heat until the spots are clearly visible. The test sample chromatogram should show spots of the same color at the corresponding positions as the reference medicinal material chromatogram.

[0166] E. Identification of Citrus aurantium:

[0167] Preparation of the test solution: Take 4g of the tablet, grind it into a fine powder, add 30ml of 100v / v% methanol, sonicate for 10 minutes, cool, filter, evaporate the filtrate to dryness, add 2ml of methanol to dissolve the residue to obtain the test solution.

[0168] Preparation of reference solutions: Take naringin reference standard and neohesperidin reference standard, add 100 v / v% methanol to prepare solutions containing 2 mg per 1 ml, and use them as reference solutions;

[0169] Citrus aurantium negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the formula does not contain Citrus aurantium;

[0170] Identification: Perform thin-layer chromatography (TLC). Apply 1 μl each of the test solution and the reference solution to the same silica gel G TLC plate. Develop using the lower layer of a chloroform-methanol-water solution (10:10:1, v / v) as the developing solvent. Develop to a distance of 8-20 cm. Remove the plate, air dry, spray with 3% aluminum chloride ethanol solution, heat at 105°C for approximately 5 minutes, and examine under ultraviolet light (365 nm). The test sample chromatogram should show fluorescent spots of the same color at the corresponding positions as the reference sample chromatogram.

[0171] F. Identification of Paeonia lactiflora:

[0172] Preparation of the test solution: Take 4g of the Chinese herbal composition, add 30ml of ethanol, shake for 20 minutes, filter, evaporate the filtrate to dryness, add 1ml of ethanol to dissolve the residue to obtain the test solution;

[0173] Preparation of reference solution: Take paeoniflorin reference standard and add ethanol to prepare a solution containing 1 mg per 1 ml, which is used as the reference solution;

[0174] Paeonia lactiflora negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the prescription does not contain Paeonia lactiflora;

[0175] Identification: Perform thin-layer chromatography. Apply 5 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Develop the plate using a chloroform-ethyl acetate-methanol-formic acid solution (v / v) of 50:2:15:0.1. Remove the plate, air dry, spray with 5% vanillin-sulfuric acid-ethanol solution, and heat until the spots are clearly visible. The test sample chromatogram should show spots of the same color at the corresponding positions as the reference sample chromatogram.

[0176] G. Identification of Coptis chinensis:

[0177] Preparation of the test solution: Take 4g of the Chinese herbal composition, add 30ml of methanol, sonicate for 5 minutes, filter, and take the filtrate to obtain the test solution;

[0178] Preparation of reference solution: Take berberine hydrochloride reference standard and add methanol to prepare a solution containing 2 mg per 1 ml, which is used as the reference solution;

[0179] Coptis chinensis negative sample solution: prepared according to the preparation method of the test solution, except that the prescription does not contain Coptis chinensis;

[0180] Identification: Perform thin-layer chromatography (TLC). Apply 10 μl each of the test solution and the reference solution to the same silica gel G TLC plate. Use a cyclohexane-ethyl acetate-isopropanol-methanol-water-triethylamine solution (v / v) as the developing solvent. Place the plate in a developing tank pre-saturated with concentrated ammonia solution for 20 minutes. Develop, remove, air-dry, and examine under ultraviolet light (365 nm). The test sample chromatogram should show fluorescent spots of the same color at the corresponding positions as the reference sample chromatogram.

[0181] Example 9

[0182] This embodiment provides a method for quality testing of a traditional Chinese medicine composition for strengthening the spleen and stomach, comprising the following steps: the traditional Chinese medicine composition for strengthening the spleen and stomach is prepared in Example 2;

[0183] A. Determination of Astragaloside A content

[0184] Preparation of the test solution: Accurately weigh approximately 5 g of the sample from the content variation section of this product, place it in a stoppered conical flask, add 20 ml of 70% methanol solution, heat under reflux for 0.1 hours, filter, evaporate the filtrate to dryness, add 10 ml of water to the residue, heat gently to dissolve, extract with water-saturated n-butanol by shaking 5 times, 10 ml each time, combine the n-butanol extracts, wash thoroughly twice with ammonia solution, 40 ml each time, discard the ammonia solution, evaporate the n-butanol extract to dryness, dissolve the residue in methanol, transfer to a 50 ml volumetric flask, dilute with methanol to the mark, and shake well to obtain the test solution.

[0185] Preparation of reference solution: Take an appropriate amount of astragaloside A reference standard, accurately weigh it, and add methanol solution to prepare a solution containing 0.5 mg per ml.

[0186] Chromatographic conditions: Octadecylsilane-bonded silica gel as the stationary phase; acetonitrile-water (32:68) as the mobile phase; evaporative light scattering detector; drift tube temperature 100℃; gas pressure 400 kPa. The theoretical plate number, calculated based on the astragaloside A peak, should be no less than 5000.

[0187] Assay: Accurately pipette 10 μl and 20 μl of the reference solution and 20 μl of the test solution into the liquid chromatograph, respectively, and determine the result using the logarithmic equation of the external standard two-point method.

[0188] Each sachet of this product contains astragalus root extract as astragaloside A (C). 41 H 68 O 14 The total amount should not be less than 0.5mg.

[0189] B. Determination of the content of paeoniflorin and naringin

[0190] Preparation of the test solution: Take the granules prepared above, weigh them accurately, add 100 mL of 40 v / v% methanol accurately, stopper tightly, weigh them, reflux in a water bath for 3 hours, cool, weigh them again, make up the lost weight with 50 v / v% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0191] Preparation of reference solution: Take appropriate amounts of paeoniflorin and naringin reference standards, accurately weigh them, and add 40 v / v% methanol solution to prepare a solution containing 70 μg of paeoniflorin and 80 μg of naringin per 1 mL.

[0192] Chromatographic conditions: The determination was performed using high-performance liquid chromatography (HPLC), with octadecylsilane-bonded silica gel as the packing material (Agilent Zorbax SB-C18 column (4.6 mm × 250 mm, 5 μm)); acetonitrile was used as mobile phase A, and 0.1% phosphoric acid solution was used as mobile phase B, with gradient elution: 0–9 min, 15 v / v% A; 9–30 min, 15 v / v% A → 22 v / v% A; 30–33 min, 22 v / v% A → 50 v / v% A; 33–35 min, 50 v / v% A; detection wavelengths were 230 nm and 283 nm; the theoretical plate number, calculated based on naringin, should not be less than 5000.

[0193] Determination method: Accurately pipette 20 μl each of paeoniflorin reference solution, naringin reference solution and test solution into the liquid chromatograph and determine the result.

[0194] Each sachet of this product contains naringin (C) from citrus aurantium. 27 H 32 O 14 The total dose shall not be less than 23.5 mg.

[0195] Each packet of this product contains paeoniflorin (C) from paeoniflorin. 23 H 28 O 11 The total dose shall not be less than 18.1 mg.

[0196] C. Identification of Angelica sinensis:

[0197] Preparation of test solution: Take 4g of the particles, add 40ml of ether, sonicate (power 250W, frequency 40kHz) for 30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 5ml of ethanol to obtain the test solution.

[0198] Preparation of reference drug solution: Take 2g of Angelica sinensis reference drug and prepare the reference drug solution using the same method;

[0199] Angelica sinensis negative sample solution: prepared according to the preparation method of the test solution, the only difference being that the prescription does not contain Angelica sinensis;

[0200] Identification: Perform thin-layer chromatography. Apply 10 μl of each of the above solutions separately to the same silica gel G thin-layer plate. Develop the plate using n-hexane-ethyl acetate (5:1, v / v) as the developing solvent. Remove the plate, air-dry it, and examine it under ultraviolet light (365 nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material.

[0201] D. Identification of Atractylodes macrocephala:

[0202] Preparation of the test solution: Take 0.5g of the capsule, add 5ml of n-hexane, sonicate for 10 minutes, cool, filter, and take the filtrate to obtain the test solution;

[0203] Preparation of the reference herb solution: Take 1g of Atractylodes macrocephala reference herb and prepare the reference herb solution according to the preparation method of the test sample;

[0204] Atractylodes macrocephala negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the prescription does not contain Atractylodes macrocephala;

[0205] Identification: Perform thin-layer chromatography. Apply 1 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Develop the plate using a 20:5 (v / v) petroleum ether (60-90℃)-ethyl acetate solution as the developing solvent. Remove the plate, air dry, spray with 5% vanillin-sulfuric acid solution, and heat until the spots are clearly visible. The test sample chromatogram should show spots of the same color at the corresponding positions as the reference medicinal material chromatogram.

[0206] E. Identification of Citrus aurantium:

[0207] Preparation of the test solution: Take 0.5g of the tablet, grind it into a fine powder, add 5ml of 40v / v% methanol, sonicate for 60 minutes, cool, filter, evaporate the filtrate to dryness, add 2ml of methanol to dissolve the residue to obtain the test solution.

[0208] Preparation of reference solutions: Naringin reference standard and neohesperidin reference standard were prepared by adding 40 v / v% methanol to prepare solutions containing 0.2 mg per ml, which were used as reference solutions.

[0209] Citrus aurantium negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the formula does not contain Citrus aurantium;

[0210] Identification: Perform thin-layer chromatography (TLC). Apply 8 μl each of the test solution and the reference solution to the same silica gel G TLC plate. Develop using the lower layer of a chloroform-methanol-water solution (15:2:4 v / v) as the developing solvent. Develop to a distance of 8-20 cm. Remove the plate, air dry, spray with 3% aluminum chloride ethanol solution, heat at 105°C for approximately 5 minutes, and examine under ultraviolet light (365 nm). The test sample chromatogram should show fluorescent spots of the same color at the corresponding positions as the reference sample chromatogram.

[0211] F. Identification of Paeonia lactiflora:

[0212] Preparation of the test solution: Take 0.5g of the Chinese herbal composition, add 5ml of 40v / v% ethanol, shake for 5 minutes, filter, evaporate the filtrate to dryness, add 1ml of 40v / v% ethanol to dissolve the residue to obtain the test solution;

[0213] Preparation of reference solution: Take paeoniflorin reference standard and add 40 v / v ethanol to prepare a solution containing 0.2 mg per 1 ml, which is used as the reference solution;

[0214] Paeonia lactiflora negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the prescription does not contain Paeonia lactiflora;

[0215] Identification: Perform thin-layer chromatography. Apply 5 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Develop the plate using a chloroform-ethyl acetate-methanol-formic acid solution (20:10:5:1, v / v). Remove the plate, air dry, spray with 5% vanillin-sulfuric acid-ethanol solution, and heat until the spots are clearly visible. The test sample chromatogram should show spots of the same color at the corresponding positions as the reference sample chromatogram.

[0216] G. Identification of Coptis chinensis:

[0217] Preparation of the test solution: Take 0.5g of the traditional Chinese medicine composition, add 5ml of 40v / v% methanol solution, sonicate for 20 minutes, filter, and take the filtrate to obtain the test solution;

[0218] Preparation of reference solution: Take berberine hydrochloride reference standard and add 40 v / v% methanol solution to prepare a solution containing 0.2 mg per ml, which is used as the reference solution;

[0219] Coptis chinensis negative sample solution: prepared according to the preparation method of the test solution, except that the prescription does not contain Coptis chinensis;

[0220] Identification: Perform thin-layer chromatography. Apply 10 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Use a cyclohexane-ethyl acetate-isopropanol-methanol-water-triethylamine solution (v / v) as the developing solvent. Develop the plate in a developing tank pre-saturated with concentrated ammonia solution for 20 minutes. Remove the plate, air-dry it, and examine it under ultraviolet light (365 nm). The test sample chromatogram should show fluorescent spots of the same color at the corresponding positions as the reference sample chromatogram.

[0221] Example 10

[0222] This embodiment provides a quality testing method for a traditional Chinese medicine composition for strengthening the spleen and stomach, comprising the following steps: the traditional Chinese medicine composition for strengthening the spleen and stomach is prepared in Example 3;

[0223] A. Determination of Astragaloside A content

[0224] Preparation of the test solution: Accurately weigh approximately 5g of the sample from the "Variation in Content" section of this product, place it in a stoppered conical flask, add 150ml of 85% methanol solution, heat under reflux for 0.1 hours, filter, evaporate the filtrate to dryness, add 10ml of water to the residue, heat gently to dissolve, extract once with 50ml of water-saturated n-butanol, combine the n-butanol extracts, wash thoroughly twice with 40ml of ammonia solution each time, discard the ammonia solution, evaporate the n-butanol extract to dryness, dissolve the residue in methanol, transfer to a 2ml volumetric flask, dilute to the mark with methanol, and shake well to obtain the test solution.

[0225] Preparation of reference solution: Take an appropriate amount of astragaloside A reference standard, accurately weigh it, and add methanol solution to prepare a solution containing 0.5 mg per ml.

[0226] Chromatographic conditions: Octadecylsilane-bonded silica gel as the stationary phase; acetonitrile-water (40:60) as the mobile phase; evaporative light scattering detector; drift tube temperature 35℃; gas pressure 300 kPa. The theoretical plate number, calculated based on the astragaloside A peak, should be no less than 5000.

[0227] Assay: Accurately pipette 10 μl and 20 μl of the reference solution and 20 μl of the test solution into the liquid chromatograph, respectively, and determine the result using the logarithmic equation of the external standard two-point method.

[0228] Each sachet of this product contains astragalus root extract as astragaloside A (C). 41 H 68 O 14 The total amount should not be less than 0.5mg.

[0229] B. Determination of the content of paeoniflorin and naringin

[0230] Preparation of the test solution: Take the granules prepared above, weigh them accurately, add 10 mL of 100 v / v% methanol accurately, stopper tightly, weigh them, reflux in a water bath for 1 hour, cool, weigh them again, make up the lost weight with 100% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0231] Preparation of reference solution: Take appropriate amounts of paeoniflorin and naringin reference standards, accurately weigh them, and add 100% methanol solution to prepare a solution containing 70 μg of paeoniflorin and 80 μg of naringin per 1 mL.

[0232] Chromatographic conditions: The determination was performed using high-performance liquid chromatography (HPLC), with octadecylsilane-bonded silica gel as the stationary phase; acetonitrile as mobile phase A and 0.1% phosphoric acid solution as mobile phase B, with gradient elution: 0–9 min, 15 v / v% A; 9–30 min, 15 v / v% A → 22 v / v% A; 30–33 min, 22 v / v% A → 50 v / v% A; 33–35 min, 50 v / v% A; detection wavelengths were 230 nm and 283 nm; the theoretical plate number, calculated based on naringin, should not be less than 5000.

[0233] The assay involves precisely injecting 2 μl each of the paeoniflorin reference solution, naringin reference solution, and test solution into a liquid chromatograph and measuring the results.

[0234] Each sachet of this product contains naringin (C) from citrus aurantium. 27 H 32 O 14 The total dose shall not be less than 23.5 mg.

[0235] Each packet of this product contains paeoniflorin (C) from paeoniflorin. 23 H 28 O 11 The total dose shall not be less than 18.1 mg.

[0236] C. Identification of Angelica sinensis:

[0237] Preparation of the test solution: Take 3g of the particles, add 30ml of ether, sonicate (power 250W, frequency 40kHz) for 10 minutes, filter, evaporate the filtrate to dryness, add 1ml of ethanol to dissolve the residue, and use it as the test solution.

[0238] Preparation of reference drug solution: Take 0.5g of Angelica sinensis reference drug and prepare the reference drug solution using the same method;

[0239] Angelica sinensis negative sample solution: prepared according to the preparation method of the test solution, the only difference being that the prescription does not contain Angelica sinensis;

[0240] Identification: Perform thin-layer chromatography (TLC). Apply 10 μl of each of the above solutions separately to the same silica gel G TLC plate. Develop with n-hexane-ethyl acetate (4:1, v / v) as the developing solvent. Remove the plate, air-dry, and examine under ultraviolet light (365 nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material. A negative control sample of Angelica sinensis was used as a reference; no interference was observed. The chromatogram is shown in [reference image]. Figure 1 As shown, 2 is a negative sample of Angelica sinensis, 1, 3, and 4 are test samples, and 5 is the reference material of Angelica sinensis.

[0241] D. Identification of Atractylodes macrocephala:

[0242] Preparation of the test solution: Take 2g of the capsule, add 10ml of n-hexane, sonicate for 15 minutes, cool, filter, and take the filtrate to obtain the test solution.

[0243] Preparation of the reference herb solution: Take 0.5g of Atractylodes macrocephala reference herb and prepare the reference herb solution according to the preparation method of the test sample;

[0244] Atractylodes macrocephala negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the prescription does not contain Atractylodes macrocephala;

[0245] Identification: Perform thin-layer chromatography. Apply 10 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Develop the plate using a petroleum ether (60-90℃)-ethyl acetate solution (v / v) at a ratio of 50:1. Remove the plate, air dry, spray with 5% vanillin-sulfuric acid solution, and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material.

[0246] E. Identification of Citrus aurantium:

[0247] Preparation of the test solution: Take 2g of the tablet, grind it into a fine powder, add 20ml of methanol, sonicate for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 2ml of methanol to dissolve the residue to obtain the test solution.

[0248] Preparation of reference solutions: Take naringin reference standard and neohesperidin reference standard, add methanol to prepare solutions containing 0.5 mg per 1 ml, and use them as reference solutions;

[0249] Citrus aurantium negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the formula does not contain Citrus aurantium;

[0250] Identification: Perform thin-layer chromatography. Apply 10 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Use the lower layer of a chloroform-methanol-water solution (13:6:2, v / v) as the developing solvent. Develop to a distance of 14 cm. Remove the plate, air dry, spray with 3% aluminum trichloride ethanol solution, heat at 105°C for about 5 minutes, and examine under ultraviolet light (365 nm). In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatogram of the reference sample.

[0251] F. Identification of Paeonia lactiflora:

[0252] Preparation of the test solution: Take 2g of the Chinese herbal composition, add 20ml of ethanol, shake for 5 minutes, filter, evaporate the filtrate to dryness, add 1ml of ethanol to dissolve the residue to obtain the test solution;

[0253] Preparation of reference solution: Take paeoniflorin reference standard and add ethanol to prepare a solution containing 2 mg per 1 ml, which is used as the reference solution;

[0254] Paeonia lactiflora negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the prescription does not contain Paeonia lactiflora;

[0255] Identification: Perform thin-layer chromatography. Apply 5 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Use a chloroform-ethyl acetate-methanol-formic acid solution (v / v) of 40:5:10:0.2 as the developing solvent. Develop, remove, and air-dry. Spray with 5% vanillin-sulfuric acid-ethanol solution and heat until the spots are clearly visible. The test sample chromatogram should show spots of the same color at the corresponding positions as the reference sample chromatogram.

[0256] G. Identification of Coptis chinensis:

[0257] Preparation of the test solution: Take 2g of the Chinese herbal composition, add 25ml of methanol, sonicate for 30 minutes, filter, and take the filtrate to obtain the test solution;

[0258] Preparation of reference solution: Take berberine hydrochloride reference standard and add methanol to prepare a solution containing 0.5 mg per ml, which is used as the reference solution;

[0259] Coptis chinensis negative sample solution: prepared according to the preparation method of the test solution, except that the prescription does not contain Coptis chinensis;

[0260] Identification: Perform thin-layer chromatography. Apply 10 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Use a cyclohexane-ethyl acetate-isopropanol-methanol-water-triethylamine solution (v / v) as the developing solvent. Develop the plate in a developing tank pre-saturated with concentrated ammonia solution for 20 minutes. Remove the plate, air-dry it, and examine it under ultraviolet light (365 nm). The test sample chromatogram should show fluorescent spots of the same color at the corresponding positions as the reference sample chromatogram.

[0261] Example 11

[0262] The difference between this embodiment and Example 8 is that it includes the following steps: the spleen-strengthening and stomach-nourishing traditional Chinese medicine composition is prepared in Example 4;

[0263] D. Identification of Atractylodes macrocephala:

[0264] Preparation of the test solution: Take 2g of the capsule, add 10ml of n-hexane, sonicate for 15 minutes, cool, filter, and take the filtrate to obtain the test solution.

[0265] Preparation of control medicinal material solution: Take 0.5 g of Atractylodes macrocephala Koidz. control medicinal material, and prepare the control medicinal material solution according to the preparation method of the test sample;

[0266] Atractylodes macrocephala Koidz. negative sample solution: Prepared according to the preparation method of the test sample solution, the difference is only that Atractylodes macrocephala Koidz. is not included in the prescription;

[0267] Identification: According to the thin-layer chromatography test, absorb 10 μl of each of the above-mentioned test sample solution and reference substance solution, and spot them on the same silica gel G thin-layer plate respectively. Use a petroleum ether (60-90 °C)-ethyl acetate solution with a volume ratio of 50:1 as the developing agent, develop, take out, dry in air, spray with 5% vanillin sulfuric acid solution, and heat until the spots are clearly colored. In the test sample chromatogram, at the corresponding position of the control medicinal material chromatogram, there are spots of the same color; after being compared with the Atractylodes macrocephala Koidz. negative sample, the negative sample has no interference. The chromatogram is shown in Figure 2 as shown, where 5 is the Atractylodes macrocephala Koidz. negative sample, 2, 3, and 4 are test sample samples, and 1 is the Atractylodes macrocephala Koidz. control medicinal material.

[0268] Example 12

[0269] The difference between this example and Example 8 is that the traditional Chinese medicine composition for invigorating the spleen and regulating the stomach is prepared in Example 5;

[0270] E. Identification of Aurantii Fructus Immaturus:

[0271] Preparation of test sample solution: Take 2 g of the tablets, grind them finely, add 20 ml of methanol, ultrasonically treat for 30 minutes, let it cool, filter, evaporate the filtrate to dryness, dissolve the residue in 2 ml of methanol to obtain the test sample solution;

[0272] Preparation of reference substance solution: Take naringin reference substance and neohesperidin reference substance, and make solutions containing 0.5 mg per 1 ml with methanol respectively as the reference substance solution;

[0273] Aurantii Fructus Immaturus negative sample solution: Prepared according to the preparation method of the test sample solution, the difference is only that Aurantii Fructus Immaturus is not included in the prescription;

[0274] Identification: According to the thin-layer chromatography test, absorb 10 μl of each of the above-mentioned test sample solution and reference substance solution, and spot them on the same silica gel G thin-layer plate respectively. Use the lower layer solution of chloroform-methanol-water with a volume ratio of 13:6:2 as the developing agent, develop, the developing distance is 14 cm, take out, dry in air, spray with 3% aluminum trichloride ethanol solution, heat at 105 °C for about 5 minutes, and examine under an ultraviolet lamp (365 nm). In the test sample chromatogram, at the corresponding position of the reference substance chromatogram, there are fluorescent spots of the same color; after being compared with the Aurantii Fructus Immaturus negative sample, the negative sample has no interference. The chromatogram is shown in Figure 3 as shown, where 1 is the neohesperidin reference substance, 2 is the naringin reference substance, 3, 4, and 5 are test sample samples, and 6 is the Aurantii Fructus Immaturus negative sample.

[0275] Example 13

[0276] The difference between this embodiment and Example 8 is that the spleen-strengthening and stomach-nourishing traditional Chinese medicine composition is prepared in Example 6;

[0277] F. Identification of Paeonia lactiflora:

[0278] Preparation of the test solution: Take 2g of the Chinese herbal composition, add 20ml of ethanol, shake for 5 minutes, filter, evaporate the filtrate to dryness, add 1ml of ethanol to dissolve the residue to obtain the test solution;

[0279] Preparation of reference solution: Take paeoniflorin reference standard and add ethanol to prepare a solution containing 2 mg per 1 ml, which is used as the reference solution;

[0280] Paeonia lactiflora negative sample solution: prepared according to the preparation method of the test sample solution, the only difference being that the prescription does not contain Paeonia lactiflora;

[0281] Identification: Perform thin-layer chromatography (TLC). Apply 5 μl each of the test solution and the reference solution to the same silica gel G TLC plate. Develop the plate using a chloroform-ethyl acetate-methanol-formic acid solution (40:5:10:0.2 v / v). Remove the plate, air dry, spray with 5% vanillin-sulfuric acid-ethanol solution, and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference sample. A negative control sample (Paeonia lactiflora) was used as a reference; no interference was observed. The chromatogram is shown in [reference image]. Figure 4 As shown, 1 is paeoniflorin reference standard, 2, 3, and 4 are test samples, and 5 is a negative sample of Paeonia lactiflora.

[0282] Example 14

[0283] The difference between this embodiment and Example 8 is that the spleen-strengthening and stomach-nourishing traditional Chinese medicine composition is prepared in Example 7;

[0284] G. Identification of Coptis chinensis:

[0285] Preparation of the test solution: Take 2g of the Chinese herbal composition, add 25ml of methanol, sonicate for 30 minutes, filter, and take the filtrate to obtain the test solution;

[0286] Preparation of reference solution: Take berberine hydrochloride reference standard and add methanol to prepare a solution containing 0.5 mg per ml, which is used as the reference solution;

[0287] Coptis chinensis negative sample solution: prepared according to the preparation method of the test solution, except that the prescription does not contain Coptis chinensis;

[0288] Identification: According to the thin-layer chromatography test, 10 μl of the above-mentioned test solution and reference solution were respectively pipetted and spotted on the same silica gel G thin-layer plate. A solution of cyclohexane - ethyl acetate - isopropanol - methanol - water - triethylamine with a volume ratio of 3:3.5:1:1.5:0.5:1 was used as the developing agent. It was placed in a developing tank pre-saturated with concentrated ammonia test solution for 20 minutes, developed, taken out, air-dried, and inspected under an ultraviolet lamp (365 nm). In the chromatogram of the test sample, at the position corresponding to the chromatogram of the reference substance, fluorescent spots of the same color appeared. After being compared with the negative sample of Coptis chinensis, the negative sample showed no interference. The chromatogram is shown in Figure 5 as follows, where 1 is the reference substance of berberine hydrochloride, 2, 3, and 4 are the test samples, and 5 is the negative sample of Coptis chinensis.

[0289] Experimental Example 1 Effects of the traditional Chinese medicine composition for invigorating the spleen and regulating the stomach on chronic gastritis rats induced by deoxycholic acid and alcohol

[0290] I. Experimental materials

[0291] Animals: SD rats, SPF grade, half male and half female, weighing 180 - 200 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License number: SCXK (Beijing) 2012 - 0001. Certificate number: 11400700125018. The experimental animals were housed in an SPF-grade animal room, illuminated by fluorescent lamps, with a 12-hour light-dark cycle, free access to water and food, at a temperature of 20°C - 26°C, and a humidity of 40 - 70% RH.

[0292] Drugs: Traditional Chinese medicine composition: Prepared in Example 1.

[0293] Hydrotalcite tablets: Bayer Healthcare Co., Ltd., batch number: 18055.

[0294] Jianwei Xiaoyan Granules: Shandong Buchang Pharmaceutical Co., Ltd., batch number: 151224.

[0295] II. Experimental methods

[0296] SD rats, half male and half female, 112 in total, were adaptively fed for 7 days and randomly divided into 2 groups: 16 rats in the blank group and 96 rats in the model group.

[0297] Rats in the model group were administered 60% ethanol every 5 days for the first month and every 7 days for the second month, 2 ml per rat each time. During this period, they were given free access to 20 mmol / L sodium deoxycholate solution (pH 7.0-7.8) instead of water. In the third month, they were given 30% ethanol solution and 20 mmol / L sodium deoxycholate solution, alternating every 3 days, with free access to the latter. Rats in the control group drank tap water normally without special treatment. After modeling, the rats in the model group were randomly divided into a model group, an aluminum magnesium carbonate tablet group, a stomach-strengthening and anti-inflammatory granule group, and high, medium, and low dose groups of the traditional Chinese medicine composition from Example 1. Each group of rats was given the corresponding test drug once daily for 30 days; the control group and model group were given the same volume of distilled water by gavage. Specific administration details are shown in Table 8 below.

[0298] Table 2. Dosage and drug concentration of each group of rats with chronic gastritis

[0299]

[0300] Before use, the drug should be prepared with distilled water to the required concentration.

[0301] Rats were weighed weekly during modeling and drug administration to observe changes in body weight. The day after the last drug administration, rats were fasted for 24 hours. Blood was collected from the abdominal aorta under anesthesia, and serum was obtained by centrifugation. Serum levels of inflammatory factors IL-8, PGE2, and gastrin were measured. The cardia and pylorus were ligated, and the entire stomach was harvested. Surface blood was washed away with distilled water, and the stomach was blotted dry with filter paper. The stomach was placed in a culture dish approximately 5 cm in diameter. The stomach cavity was dissected along the lesser curvature, containing a small amount of gastric juice (approximately 0.5 ml). The stomach wall was inverted, and 4 ml of distilled water was added for rinsing. The rinsing solution was transferred to a 5 ml graduated test tube and diluted to 5 ml with distilled water (approximately a 10-fold dilution of the gastric juice). The pH of the gastric juice was measured using a DELTA320 pH meter. Take 1 ml of diluted gastric juice from each rat, add 1 drop of 0.1% phenol red as an indicator, and titrate with 0.002 mol / L NaOH. Carefully titrate until the solution color changes from yellow to red, which is the amount of NaOH consumed by the free acid. Record the amount of NaOH and calculate the free acidity of the gastric juice (mmol / L) according to the formula mol × volume = mol1 × volume1.

[0302] The gastric juice of each rat was diluted and the content of free mucus in the gastric juice was determined using Alcian Blue.

[0303] Separately, gastric juice from each rat was diluted, and pepsin activity was measured using a pepsin kit.

[0304] After cleaning, the stomach was fixed with 10% formaldehyde. A 3mm × 10mm section of the full-thickness stomach wall was cut off along the lesser curvature of the stomach, from the foregut to the pylorus, and stained with hematoxylin and eosin (HE) to observe the pathological changes in the gastric mucosa.

[0305] Statistical methods

[0306] Experimental data are expressed as mean ± standard deviation. The data were first tested for homogeneity of variance. For homogeneous variances, One-Way ANOVA was used for comparisons between groups; for unequal variances, the t' test was used for comparisons between groups.

[0307] III. Experimental Results

[0308] (1) General condition of rats in each group

[0309] The control group rats had glossy white fur, bright eyes, were more active and agile, and had a larger appetite, which increased with weight gain in the early stages and remained relatively stable in the later stages. Their feces were granular, their tails were pale red, and there were no abnormal secretions from their noses, eyes, or mouths. Their breathing was even. The model group rats generally experienced weight loss during the modeling process, reduced spontaneous activity, huddled together, arched backs, decreased appetite, lethargy, and their fur became dark, yellowish, sparse, rough, and prone to shedding. They were smaller in size and gradually became emaciated. The drug-treated groups showed a significant increase in weight and a marked improvement in symptoms compared to the model group after drug administration. The changes in rat weight after modeling and after drug administration are shown in the table below.

[0310] Table 3. Effects of traditional Chinese medicine composition on body weight in rats with chronic gastritis.

[0311]

[0312] Table 3. Effects of the traditional Chinese medicine composition on the body weight of rats with chronic gastritis.

[0313]

[0314]

[0315] Compared with the blank group: # P<0.05, ## P<0.01; Compared with the model group: * P<0.05, ** P<0.01.

[0316] (2) Effects on gastric juice pH, free acidity and pepsin activity in rats with chronic gastritis

[0317] The results showed that after modeling, the pH value of gastric juice in the model group rats increased, the free acidity in gastric juice decreased, and the pepsin activity in gastric juice increased, which were significantly different from those in the blank group (p<0.01, p<0.01, p<0.05, respectively), indicating that the gastric mucosa of the model rats was damaged.

[0318] Compared with the model group, the aluminum magnesium carbonate tablet group, the stomach-strengthening and anti-inflammatory granule group, and the high, medium, and low dose groups of the traditional Chinese medicine composition in Example 1 all significantly reduced the gastric juice pH value of model rats (p<0.05, 0.05, 0.01, 0.05, 0.05, respectively) and increased the free acidity of gastric juice in model rats (all p<0.05). Compared with the model group, the high, medium, and low dose groups of the traditional Chinese medicine composition all significantly reduced the pepsin activity in the gastric juice of model rats (p<0.05, 0.01, 0.01, respectively). This suggests that the drug has a significant effect on improving the symptoms of chronic gastritis. The results are shown in the table below.

[0319] Table 4. Effects of traditional Chinese medicine composition on gastric juice pH, free acidity, and pepsin activity in rats with chronic gastritis

[0320]

[0321]

[0322] Compared with the blank group: # P<0.05, ## P<0.01; Compared with the model group: * P<0.05, ** P<0.01.

[0323] (3) Effect on the content of free mucus in gastric juice of rats with chronic gastritis

[0324] The results showed that after modeling, the content of free mucus in the gastric juice of rats in the model group increased significantly compared with that in the blank group (p<0.01), indicating that the gastric mucosa-bicarbonate barrier of the model rats was damaged.

[0325] Compared with the model group, the aluminum magnesium carbonate tablet group, the stomach-strengthening and anti-inflammatory granule group, and the high, medium, and low dose groups of the traditional Chinese medicine composition all significantly reduced the free mucus content in the gastric juice of model rats (p<0.01, 0.01, 0.01, 0.01, and 0.05, respectively). This suggests that the drugs have a protective effect on the gastric mucosa-bicarbonate barrier in model rats. The results are shown in the table below.

[0326] Table 5. Effects of traditional Chinese medicine composition on the content of free mucus in gastric juice of rats with chronic gastritis

[0327]

[0328] Compared with the blank group: # P<0.05, ## P<0.01; Compared with the model group: * P<0.05, ** P<0.01.

[0329] (4) Effects on the serum inflammatory factor IL-8 levels in rats with chronic gastritis

[0330] The results showed that after modeling, the serum IL-8 level in the model group rats was significantly increased, and there was a significant difference compared with the blank group (p<0.01), indicating that an inflammatory response occurred in the model rats.

[0331] Compared with the model group, the aluminum magnesium carbonate tablet group, the stomach-strengthening and anti-inflammatory granule group, and the high, medium, and low dose groups of the traditional Chinese medicine composition all significantly reduced the serum IL-8 level in model rats (p<0.05, p<0.01, p<0.01, p<0.01, p<0.01, respectively). This suggests that the drugs have a good inhibitory effect on the secretion of serum inflammatory factors in chronic gastritis. The results are shown in the table below.

[0332] Table 6. Effects of traditional Chinese medicine composition on serum IL-8 levels in rats with chronic gastritis

[0333]

[0334]

[0335] Compared with the blank group: # P<0.05, ## P<0.01; Compared with the model group: * P<0.05, ** P<0.01.

[0336] (5) Effect on serum gastrin levels in rats with chronic gastritis

[0337] The results showed that, compared with the blank group, the serum gastrin level of the model group rats was significantly increased (p<0.01), indicating that the model rats had hypersecretion of gastrin.

[0338] Compared with the model group, both the Jianwei Xiaoyan Granules group and the high-dose traditional Chinese medicine composition group significantly reduced serum gastrin levels in model rats (p<0.05 and p<0.01, respectively). This suggests that the drugs have a significant anti-gastrin secretion effect. The results are shown in the table below.

[0339] Table 7. Effects of Traditional Chinese Medicine Composition on Serum Gastrin Content in Rats with Chronic Gastritis

[0340]

[0341] Compared with the blank group: # P<0.05, ## P<0.01; Compared with the model group: * P<0.05, ** P<0.01.

[0342] (6) Effect on serum PGE2 levels in rats with chronic gastritis

[0343] The results showed that, compared with the control group, the serum PGE2 level in the model group rats was significantly reduced (p<0.01).

[0344] Compared with the model group, the aluminum magnesium carbonate tablet group, the stomach-strengthening and anti-inflammatory granule group, and the high, medium, and low dose groups of the traditional Chinese medicine composition all significantly increased the serum PGE2 level in the model rats (all p<0.01). This suggests that the traditional Chinese medicine composition has a protective effect on the gastric mucosa. The results are shown in the table below.

[0345] Table 8. Effects of traditional Chinese medicine composition on serum PGE2 levels in rats with chronic gastritis

[0346]

[0347] Compared with the blank group: # P<0.05, ## P<0.01; Compared with the model group: * P<0.05, ** P<0.01.

[0348] (7) Effects on the pathological changes of gastric tissue in rats with chronic gastritis

[0349] Macroscopic observation of gastric tissue morphology: blank group (e.g.) Figure 12 (As shown) The rat gastric mucosa was pale red, smooth, and glossy, with visible mucus covering it. It was elastic, without a granular feel, and the mucosal folds were intact and of moderate thickness. The gastric mucosa of the model group rats was pale white and dull. The folds were low and disordered. Some animals had congested and edematous gastric mucosa with scattered bleeding points. A few animals had superficial erosions in the mucosa, and the posterior wall of the gastric body mucosa was thickened and rough. The aluminum magnesium carbonate tablet group, the stomach-strengthening and anti-inflammatory granule group, and the high, medium, and low dose groups of the traditional Chinese medicine composition showed better improvement in gastric tissue lesions compared to the model group.

[0350] Pathological histological examination results showed that the gastric mucosa of rats in the model group was congested and edematous, with degeneration, necrosis and shedding of mucosal epithelial cells. At the same time, inflammatory cells were common in the submucosa and lamina propria and migrated into the mucosal layer. Inflammatory cell infiltration (mainly lymphocytes and eosinophils, with a small number of neutrophils) was observed in the lamina propria. The gastric glands were disordered or unevenly distributed.

[0351] The aluminum magnesium carbonate tablet group, the stomach-strengthening and anti-inflammatory granule group, and the high, medium, and low dose groups of the traditional Chinese medicine composition all showed varying degrees of ablation on the pathological changes in the gastric mucosa of the above-mentioned model rats. The pathological changes in the gastric mucosa of the aluminum magnesium carbonate tablet group were significantly improved compared to the model group. In a few rats, small patches of mucosal tissue necrosis and a small amount of exudate were observed in localized areas of the gastric mucosa, but the lesions were superficial, with mild inflammatory cell infiltration in the submucosa and lamina propria. Glandular changes were significantly reduced compared to the model group. The pathological changes in the gastric mucosa of the stomach-strengthening and anti-inflammatory granule group were also improved compared to the model group. In a few rats, small patches of mucosal tissue necrosis and a small amount of exudate were observed in localized areas of the gastric mucosa, but the lesions were superficial, with mild inflammatory cell infiltration in the submucosa and lamina propria. Glandular changes were significantly reduced compared to the model group. The high dose group of the traditional Chinese medicine composition significantly reduced the severity of gastric mucosal lesions, and occasionally, small patches of gastric mucosal lesions were observed in rats. The gastric mucosa showed degeneration and necrosis, but this was significantly less severe than in the model group. The number of inflammatory cells in the lamina propria of the gastric mucosa decreased, and the gastric glands were arranged more neatly and evenly with more regular ductal structures. The medium-dose group of the traditional Chinese medicine composition showed a significant reduction in the severity of gastric mucosal lesions. In some rats, the gastric mucosa showed degeneration and necrosis, but this was significantly less severe than in the model group. The number of inflammatory cells in the gastric mucosa and lamina propria decreased, and the gastric glands were arranged more neatly and evenly with more regular ductal structures. The low-dose group of the traditional Chinese medicine composition showed a significant reduction in the severity of gastric mucosal lesions. It was observed that the gastric mucosa showed degeneration and necrosis, a decrease in the number of inflammatory cells in the lamina propria of the gastric mucosa, and the gastric glands were arranged more neatly and evenly with more regular ductal structures. This significantly reduced the pathological morphology score of the gastric mucosa, showing a significant difference compared to the model group (p<0.05, p<0.05, p<0.01, p<0.01, p<0.05, respectively). This suggests that the extract of the traditional Chinese medicine composition can significantly improve the pathological and histological changes of chronic gastric mucosal injury induced by deoxycholic acid and alcohol in rats, and has a good effect on improving gastric mucosal lesions in chronic gastritis.

[0352] Experimental Example 2: Inhibitory Effect of a Traditional Chinese Medicine Composition for Strengthening the Spleen and Stomach on the Growth of Helicobacter pylori in In Vitro Culture 1. Materials and Methods

[0353] (1) Resuscitate the strains, take FC13061 (NCTC11637) and FC13065 (SS2000) from the strain bank, take a sterile 10um inoculation loop to dip into the bacterial solution, streak on a Columbia blood plate containing 5% sheep blood, and incubate at 36℃ for 48h in a microaerophilic environment.

[0354] (2) Pick the first generation plate colonies, streak them on a Columbia blood agar plate containing 5% sheep blood, and incubate at 36 degrees Celsius for 48 hours to complete the second generation culture;

[0355] (3) Perform API reagent strip identification and mass spectrometry incubation:

[0356] 1) API reagent strip identification: Take out the API CAMPY reagent strip and the accompanying 3ml bottle of physiological saline. Pick colonies and prepare a bacterial suspension with a McFarland turbidity of 6 (after dilution 3 times, the McFarland turbidity is 2). According to the instructions, add the bacterial suspension (100ul / well) to the URE to PAL wells in Part 1 and 150ul / well to the H2S wells in Part 2. Seal the URE with mineral oil. Place the reagent strip in Part 1 in an incubator, cover, and incubate at 36℃ for 24 hours under microaerophilic conditions. Take the remaining bacterial suspension with a McFarland turbidity of 6 and add it to the accompanying culture medium. Mix well. In Part 2... Add the prepared bacterial suspension (150 μL / well) to the GLU to ERO of the reagent strip. Place the reagent strip in an incubator, cover, and incubate at 36°C for 24 hours under microaerophilic conditions. According to the instructions, add one drop each of NIT1 and NIT2 to the NIT wells, and one drop each of NIN, GGT, PyrA, ArgA, AspA, and PAL to the HIP wells, along with one drop of FB. After waiting 5 minutes, record the positive or negative result based on the color change (the second part of the reagent strip is negative after 24 hours; continue incubation for 48 hours). Input the results into the API web database to obtain the final result.

[0357] 2) Pick colonies and perform formic acid extraction (dissolve colonies in 300ul deionized water, add 900ul anhydrous ethanol, centrifuge at 13000rpm for 2min, discard the supernatant and completely remove the ethanol, add 50ul 70% formic acid to fully dissolve the precipitate after centrifugation, then add 50ul acetonitrile, mix thoroughly, centrifuge at 13000rpm for 2min, and take the supernatant as the sample). Take 1ul of the supernatant as the sample solution, and after drying, perform instrumental analysis.

[0358] (4) Determination of antibacterial activity by agar dilution method

[0359] 1) Weigh 3.3g of the herbal composition from Example 1 and dissolve it in 10ml of physiological saline (1g of herbal composition contains 3g of crude drug) to prepare a stock solution of 1g crude drug / ml, and autoclave for later use;

[0360] 2) High-pressure preparation of MH agar, and addition of sheep blood to prepare MH agar containing 5% sheep blood;

[0361] 3) Take a 50ml centrifuge tube and add 4ml, 2ml, 1ml, 0.5ml, 0.25ml, 0.125ml, 0.063ml, and 0.032ml of the drug stock solution and 4ml of physiological saline, respectively. Then add 2) blood agar to 40ml, mix well, and divide into two petri dishes (two parallel plates for one drug concentration). Let them air dry and set aside for later use. Label the plates as 0, 1, 2, 3, 4, 5, 6, 7 and NC, respectively.

[0362] 4) Select FC13061 and adjust its McFarland turbidity to 1 (equivalent to 6×).8 A bacterial suspension (cfu / ml) was prepared and then serially diluted 10-fold. 100 μL of each solution was added to each of the petri dishes in step 3), resulting in a final colony count on the order of 10^10^12 colonies per dish. 4 L-shaped rod coating;

[0363] 5) Clarithromycin was selected as a positive control. After the drug was mixed with the blood agar in 2) to 40 ml, the final concentrations were 25, 12.5, 6.25, 3.125, 1.56, and 0.78 mg / L.

[0364] 6) Process FC13065 in the same way as 1) and 5);

[0365] (5) Incubate at 36℃ under microaerophilic conditions for 2 days;

[0366] (6) Observation results.

[0367] 2. Results

[0368] (1) Both strains FC13061 (NCTC11637) and FC13065 (SS2000) were identified as Helicobacter pylori;

[0369] (2) Both strains FC13061 (NCTC11637) and FC13065 (SS2000) had colonies on plates labeled 6, 7 and NC, meaning the minimum inhibitory concentration was 5 plates.

[0370] (3) Positive controls all showed colonies on plates with concentrations of 1.56, 0.78, and 0 mg / L.

[0371] 3. Analysis

[0372] The minimum inhibitory concentration (MIC) of clarithromycin is 3.125 mg / L. On the plate marked 0, the drug concentration is 1 g crude drug / ml × (1 / 10) = 100 mg crude drug / ml. Therefore, on the plate marked 5, the drug concentration is 100 mg crude drug / ml × (1 / 32) = 3.125 mg crude drug / ml. This shows that the extract of the traditional Chinese medicine composition has a good inhibitory effect on Helicobacter pylori (NCTC11637, SS2000) cultured in vitro, and its MIC is 3.125 mg crude drug / ml.

[0373] Experimental Example 3: Effect of Traditional Chinese Medicine Composition on Painful Stimuli Response

[0374] I. Effects of Traditional Chinese Medicine Compositions on the Response of Mice to Acetic Acid Intraperitoneal Pain Stimulation

[0375] 1. Experimental materials

[0376] 1.1 Experimental Drugs

[0377] Traditional Chinese medicine composition: Preparation as described in Example 1.

[0378] Aspirin Effervescent Tablets: AstraZeneca Pharmaceuticals Co., Ltd., batch number: 1512117.

[0379] Jianwei Xiaoyan Granules: Shandong Buchang Pharmaceuticals Co., Ltd., batch number: 151224.

[0380] 1.2 Experimental reagents

[0381] Glacial acetic acid: Beijing Chemical Plant, batch number: 20150312, analytical pure.

[0382] 1.3 Experimental instruments

[0383] Haier BCD-257SL Refrigerator-Freezer, Qingdao Haier Co., Ltd.

[0384] Sartorious BT 223S Electronic Precision Balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.

[0385] Oregon Scientific WB388 Electronic Timer, Oregon Scientific Trading (Shanghai) Co., Ltd.

[0386] 1.4 Experimental animals

[0387] ICR mice, SPF grade, half male and half female, body weight 18 - 20 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License number: SCXK (Beijing) 2012 - 0001, certificate number: 11400700148304.

[0388] The experimental animals were housed in a SPF-grade animal room, illuminated by fluorescent lamps with a 12-hour light-dark cycle, and had free access to water and food. The temperature was 20℃ - 26℃, and the humidity was 40 - 70% RH.

[0389] 2. Experimental methods

[0390] SPF-grade mice, half male and half female, 72 in total. After 4 days of adaptive feeding, the mice were randomly divided into 6 groups of 12 each, namely: model group, aspirin group, jianwei xiaoyan granule group, high, medium, and low dose groups of the traditional Chinese medicine composition extract.

[0391] Each group of mice was given the test drug by gavage, once a day for 5 consecutive days. 1 hour after the last dose, 0.2 ml / animal of 0.6% glacial acetic acid normal saline solution was injected intraperitoneally into the mice, and then the number of writhing responses and the latency of writhing responses (the time from intraperitoneal injection of glacial acetic acid to the appearance of writhing responses) of the mice were observed within 15 minutes. The model group was given an equal volume of distilled water by gavage. The specific dosing conditions are shown in the following table:

[0392] Table 9. Dosing doses and drug concentration of each group of mice

[0393]

[0394] Before use, the drug should be prepared with distilled water to the required concentration.

[0395] 3. Statistical Methods

[0396] Experimental data are expressed as mean ± standard deviation. The data were first tested for homogeneity of variance. For homogeneous variances, One-Way ANOVA was used for comparisons between groups; for unequal variances, the t' test was used for comparisons between groups.

[0397] 4. Experimental Results

[0398] The experimental results showed that the aspirin group and the high, medium and low dose groups of the traditional Chinese medicine extract could reduce the number of writhing responses induced by intraperitoneal acetic acid stimulation in mice, and the differences were significant compared with the model group (all p<0.01).

[0399] The aspirin group significantly prolonged the latency of writhing response induced by intraperitoneal acetic acid stimulation in mice compared with the model group (p<0.05). The high, medium, and low dose groups of the traditional Chinese medicine composition showed a certain trend of prolonging the latency of writhing response, but there was no significant difference compared with the model group (all p>0.05). This suggests that the extract of the traditional Chinese medicine composition has an inhibitory effect on pain induced by chemical stimulation. The results are shown in the table below.

[0400] Table 10. Effects of Traditional Chinese Medicine Compositions on Acetic Acid Intraperitoneal Pain Stimulation in Mice

[0401]

[0402] Compared with the model group: * P<0.05; ** P<0.01.

[0403] II. Effects of Traditional Chinese Medicine Combinations on Hot Plate Pain Stimulation in Mice

[0404] 1. Experimental Materials

[0405] 1.1 Experimental Drugs

[0406] Traditional Chinese medicine composition: Preparation as described in Example 1.

[0407] Aspirin effervescent tablets: AstraZeneca Pharmaceutical Co., Ltd., batch number: 1512117.

[0408] Jianwei Xiaoyan Granules: Shandong Buchang Pharmaceutical Co., Ltd., Batch No.: 151224.

[0409] 1.2 Experimental Apparatus

[0410] Haier BCD-257SL Refrigerator-Freezer, Qingdao Haier Co., Ltd.

[0411] Sartorious BT 223S Electronic Precision Balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.

[0412] YLS-6B Intelligent Hot Plate, Equipment Station of Shandong Academy of Medical Sciences

[0413] 1.3 Experimental Animals

[0414] ICR mice, SPF grade, female, weighing 18 - 20 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License number: SCXK(Beijing)2012 - 0001, Certificate number: 11400700153522.

[0415] The experimental animals were housed in a SPF - grade animal room with fluorescent lighting, a 12 - h light - dark cycle, free access to water and food, a temperature of 20℃ - 26℃, and a humidity of 40 - 70%RH.

[0416] 2. Experimental Methods

[0417] 80 SPF - grade female mice were adaptively fed for 5 days, and a hot plate was used to screen qualified mice. The screening method was as follows: The hot plate temperature was adjusted to 55 ± 0.5℃. Female mice were placed on the hot plate, and the time (seconds) from when the mouse was placed on the hot plate until it licked its hind paw was used as the pain threshold of the mouse. Any mouse with a licking time of less than 5 seconds, more than 30 seconds, or one that jumped was discarded. The qualified mice were randomly divided into 6 groups according to their baseline pain thresholds, namely: model group, aspirin group, Jianwei Xiaoyan Keli group, high, medium, and low dose groups of the traditional Chinese medicine composition. Except for 10 mice in the low - dose group of the traditional Chinese medicine composition, the other groups had 11 mice each.

[0418] Each group of mice was given the test drug by gavage once a day for 5 consecutive days. After the last dose, at different time points of 0.5 h, 1.0 h, 1.5 h, 2.0 h, and 3.0 h, the mice were placed on the hot plate (55 ± 0.5℃), and the pain threshold of the mice at different time points after drug administration (the time from being placed on the hot plate until licking the hind paw) was recorded. The specific drug administration details are shown in the following table:

[0419] Table 10. Drug administration doses and preparation concentrations for each group of mice

[0420]

[0421]

[0422] Before use, the drug was prepared into the required concentration with distilled water.

[0423] 3. Statistical Methods

[0424] Experimental data are expressed as mean ± standard deviation. The data were first tested for homogeneity of variance. For homogeneous variances, One-Way ANOVA was used for comparisons between groups; for unequal variances, the t' test was used for comparisons between groups.

[0425] 4. Experimental Results

[0426] Experimental results showed that aspirin increased the pain threshold in mice, with an onset time of 0.5 hours and a duration of effect of 2.5 hours; Jianwei Xiaoyan granules increased the pain threshold in mice, with an onset time of 1.5 hours and a duration of effect of 0.5 hours; high, medium, and low doses of the traditional Chinese medicine extract all increased the pain threshold in mice, with an onset time of 1 hour and a duration of effect of 1 hour. These results suggest that the traditional Chinese medicine extract has a certain inhibitory effect on thermal pain stimulation. The results are shown in the table below.

[0427] Table 11. Effects of traditional Chinese medicine composition on pain threshold of hot plate pain stimulus in mice

[0428]

[0429] Experiment Example 4: Effect of Traditional Chinese Medicine Composition on Acute Inflammatory Response to Xylene-Induced Ear Swelling in Mice

[0430] 1. Experimental Materials

[0431] 1.1 Experimental Drugs

[0432] Traditional Chinese medicine composition: Preparation as described in Example 1.

[0433] Aspirin effervescent tablets: AstraZeneca Pharmaceutical Co., Ltd., batch number: 1512117.

[0434] Jianwei Xiaoyan Granules: Shandong Buchang Pharmaceutical Co., Ltd., Batch No.: 151224.

[0435] 1.2 Experimental Reagents

[0436] Xylene: Beijing Chemical Plant, batch number: 20140617, analytical grade.

[0437] 1.3 Experimental Apparatus

[0438] Haier BCD-257SL refrigerator and freezer, Qingdao Haier Co., Ltd.

[0439] Sartorious BT 223S electronic precision balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.

[0440] AR3130 electronic precision balance, Ohaus International Trading (Shanghai) Co., Ltd.

[0441] Hepu corneal trephine: 8 mm in diameter.

[0442] 1.4 Experimental animals

[0443] ICR mice, SPF grade, male, weighing 18 - 20 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License number: SCXK(Beijing)2012 - 0001, certificate number: 11400700148318.

[0444] The experimental animals were housed in an SPF - level animal room with fluorescent lighting, a 12 - hour light - dark cycle, free access to water and food, a temperature of 20℃ - 26℃, and a humidity of 40 - 70%RH.

[0445] 2. Experimental methods

[0446] After 3 days of adaptive feeding of the experimental animals, the mice were randomly divided into 6 groups of 12 each, namely: model group, aspirin group, Jianwei Xiaoyan Keli group, high - dose, medium - dose, and low - dose groups of the traditional Chinese medicine composition.

[0447] Each group of mice was orally administered the corresponding dose of the drug once a day for 5 consecutive days. The model group was given an equal volume of distilled water. The specific drug administration is shown in the following table:

[0448] Table 12. Drug administration doses and dispensing concentrations for each group of mice

[0449]

[0450]

[0451] Before use, the drug was prepared into the required concentration with distilled water.

[0452] 40 minutes after the last drug administration, 25 μl of xylene was evenly applied to both the front and back sides of the left ear of the mice (a total of 50 μl / left ear). One hour after the application of xylene, the mice were sacrificed, and the left and right ears were cut off. Circular ear pieces were drilled at the same position on the left and right ears using a corneal trephine with a diameter of 8 mm. The weights of the ear pieces were accurately weighed on an electronic precision balance, and the ear swelling degree and swelling rate were calculated.

[0453] Swelling degree = weight of the inflamed ear - weight of the non - inflamed ear;

[0454] Swelling rate = swelling degree / weight of the non - inflamed ear × 100%.

[0455] 3. Statistical methods

[0456] The experimental data were expressed as "mean ± standard deviation" The data were first tested for homogeneity of variance. For homogeneous variances, One-Way ANOVA was used for comparisons between groups; for unequal variances, the t' test was used for comparisons between groups.

[0457] 4. Experimental Results

[0458] The experimental results showed that the high, medium and low dose groups of the Chinese herbal composition extract could inhibit the ear swelling caused by xylene-induced acute inflammatory response in mice, and the differences were significant compared with the model group (p<0.01, p<0.05 and p<0.01, respectively).

[0459] All three doses of the traditional Chinese medicine composition reduced the ear swelling rate in mice induced by xylene-induced acute auricular inflammation, showing significant differences compared to the model group (all p < 0.01). This suggests that the traditional Chinese medicine composition has an inhibitory effect on acute inflammation. The results are shown in the table below.

[0460] Table 13. Effects of extracts from traditional Chinese medicine compositions on xylene-induced ear swelling in mice.

[0461]

[0462] Compared with the model group: * P<0.05, ** P<0.01.

[0463] Experiment Example 5: Effect of Traditional Chinese Medicine Composition on Peritoneal Capillary Permeability in Mice

[0464] 1. Experimental Materials

[0465] 1.1 Experimental Drugs

[0466] Traditional Chinese medicine composition: Preparation as described in Example 1.

[0467] Dexamethasone: Tianjin Lisheng Pharmaceutical Co., Ltd., batch number: 1407018.

[0468] Jianwei Xiaoyan Granules: Shandong Buchang Pharmaceutical Co., Ltd., Batch No.: 151224.

[0469] 1.2 Experimental Reagents

[0470] Glacial acetic acid: Beijing Chemical Plant, batch number: 20150312, analytical grade.

[0471] Evans Blue: Sinopharm Chemical Reagent Co., Ltd., Batch No.: F20040213.

[0472] 1.3 Experimental Apparatus

[0473] Haier BCD-257SL refrigerator and freezer, Qingdao Haier Co., Ltd.

[0474] Sartorius BT 223S electronic precision balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.

[0475] Epoch2 microplate reader, BioTek, USA.

[0476] 1.4 Experimental animals

[0477] ICR mice, SPF grade, half male and half female, body weight 18 - 20 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License number: SCXK (Beijing) 2012 - 0001, certificate number: 11400700152332.

[0478] The experimental animals were housed in a SPF - level animal room, illuminated by fluorescent lamps with a 12 - h light - dark cycle, with free access to water and food, at a temperature of 20℃ - 26℃ and a humidity of 40 - 70% RH.

[0479] 2. Experimental methods

[0480] After 5 days of adaptive feeding of the experimental animals, the mice were randomly divided into 6 groups of 12 each, namely: model group, dexamethasone group, Jianwei Xiaoyan Keli group, high - dose, medium - dose, and low - dose groups of the traditional Chinese medicine composition extract.

[0481] Each group of mice was given the corresponding dose of the drug by oral gavage once a day for 5 consecutive days. The model group was given an equal volume of distilled water. The specific dosing is shown in the following table:

[0482] Table 14. Dosing doses and drug - preparation concentrations for each group of mice

[0483]

[0484] Before use, the drug was prepared into the required concentration with distilled water.

[0485] Forty minutes after the last dose, 0.1 ml / 10 g of 0.5% Evans blue saline solution was injected into the tail vein of each mouse, and then 0.2 ml / rat of 0.8% glacial acetic acid saline solution was injected. Twenty minutes later, the mice were sacrificed by cervical dislocation, 4 ml of normal saline was injected intraperitoneally. After gently massaging the abdomen of the mice, the abdominal skin and muscle were cut open, 1 ml of the liquid was aspirated, centrifuged at 3000 r / min for 15 min, and the supernatant was taken for absorbance measurement at 540 nm.

[0486] 3. Statistical methods

[0487] The experimental data were expressed as "mean ± standard deviation" ". The data were first subjected to a homogeneity - of - variance test. If the variances were homogeneous, the One - Way ANOVA method was used for comparison between groups; if the variances were not homogeneous, the t’ - test was used for comparison between groups.

[0488] 4. Experimental Results

[0489] The experimental results showed that the dexamethasone group, as well as the high and medium dose groups of the traditional Chinese medicine composition, could reduce the absorbance of Evans blue exudate in the peritoneum of mice, with significant differences compared to the model group (p<0.01, p<0.01, and p<0.05, respectively). This suggests that the traditional Chinese medicine composition has a certain inhibitory effect on acetic acid-induced peritoneal capillary permeability in mice. The results are shown in the table below.

[0490] Table 15. Effects of traditional Chinese medicine composition on peritoneal capillary permeability in mice.

[0491]

[0492] Compared with the model group: * P<0.05, ** P<0.01.

[0493] Experimental Example 6: The effect of traditional Chinese medicine composition on promoting gastrointestinal motility

[0494] 1. Experimental Materials

[0495] 1.1 Experimental Drugs

[0496] Traditional Chinese medicine composition: Preparation as described in Example 1.

[0497] Domperidone: Xi'an Janssen Pharmaceutical Co., Ltd., batch number: 151215044.

[0498] Jianwei Xiaoyan Granules: Shandong Buchang Pharmaceutical Co., Ltd., Batch No.: 151224.

[0499] 1.2 Experimental Reagents

[0500] Sodium carboxymethyl cellulose: Tianjin Guangfu Fine Chemical Research Institute, batch number: November 14, 2011.

[0501] Milk powder: Shuangcheng Nestlé Co., Ltd., Production date: December 26, 2015, 13.

[0502] Fine white sugar: Beijing Sugar, Tobacco & Alcohol Group Co., Ltd., Production date: March 21, 2016.

[0503] Starch: Beijing Chudama Food Group Co., Ltd., Production Date: February 22, 2016, 12:00 PM.

[0504] Activated carbon: Sinopharm Chemical Reagent Co., Ltd., batch number: 0705250.

[0505] 1.3 Experimental Apparatus

[0506] Haier BCD-257SL Refrigerator-Freezer, Qingdao Haier Co., Ltd.

[0507] Sartorius BT 223S Electronic Precision Balance, Sartorius Scientific Instruments (Beijing) Co., Ltd.

[0508] AR3130 Electronic Precision Balance, Ohaus International Trade (Shanghai) Co., Ltd.

[0509] 1.4 Experimental Animals

[0510] ICR mice, SPF grade, half male and half female, body weight 18 - 20 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. License number: SCXK(Beijing)2012 - 0001, Certificate number: 11400700150618.

[0511] The experimental animals were housed in a SPF - level animal room, illuminated by fluorescent lamps with a 12 - h light - dark cycle, free access to drinking water and food, temperature 20℃ - 26℃, humidity 40 - 70%RH.

[0512] 2. Experimental Methods

[0513] After 4 days of adaptive feeding of the experimental animals, the mice were randomly divided into six groups of 12 each, namely: blank group, domperidone group, Jianwei Xiaoyan Keli group, high, medium, and low dose groups of the traditional Chinese medicine composition extract. Each group of mice was given the corresponding dose of the drug by oral gavage once a day for 5 consecutive days, and the model group was given an equal volume of distilled water. The specific dosing situation is shown in the following table:

[0514] Table 16. Drug Dosing Dosage and Preparation Concentration for Each Group of Mice

[0515]

[0516]

[0517] Before use, the drug was prepared into the required concentration with distilled water.

[0518] Mice in each group were administered the tested drug via gavage once daily for 5 consecutive days. One hour after the last administration, mice that had been fasted (but not water-exclusive) for 24 hours were gavage with 0.8 mL of a nutritional semi-solid paste per mouse. Twenty minutes later, the mice were euthanized by cervical dislocation. The abdomen was opened, the cardia and pylorus of the stomach were ligated, the stomach was removed, dried with filter paper, and weighed. The stomach body was then cut along the greater curvature, the stomach contents were washed away, dried, and weighed. The difference between the total stomach weight and the net stomach weight was taken as the weight of the gastric residue. The percentage of the gastric residue relative to the weight of the administered semi-solid paste was calculated as the gastric residue rate. Simultaneously, the small intestine was quickly removed, the mesentery was gently peeled off, and the intestine was laid flat on white paper. The total length from the pylorus to the ileocecal region and the distance from the pylorus to the leading edge of the black semi-solid paste were measured. The percentage of the distance from the pylorus to the leading edge of the black semi-solid paste relative to the total length from the pylorus to the ileocecal region was taken as the small intestine propulsion rate.

[0519] (Preparation of the nutritional semi-solid paste: Dissolve 10g of sodium carboxymethyl cellulose in 250mL of distilled water, then add 16g of milk powder, 8g of sugar, 8g of starch, and 2g of activated charcoal powder, and stir well. Prepare a black semi-solid paste by mixing 300mL of the solution with approximately 300g of the paste.)

[0520] 3. Statistical Methods

[0521] Experimental data are expressed as mean ± standard deviation. The data were first tested for homogeneity of variance. For homogeneous variances, One-Way ANOVA was used for comparisons between groups; for unequal variances, the t' test was used for comparisons between groups.

[0522] 4. Experimental Results

[0523] The experimental results showed that the domperidone group, the stomach-strengthening and anti-inflammatory granule group, and the high, medium, and low dose groups of the traditional Chinese medicine composition extract could all increase the small intestinal propulsion rate of mice, and there were significant differences compared with the blank group (all p<0.01), indicating that the traditional Chinese medicine composition has the function of promoting small intestinal motility.

[0524] The domperidone group, the stomach-strengthening and anti-inflammatory granule group, and the high, medium, and low dose groups of the traditional Chinese medicine composition all reduced the weight of gastric residue and the rate of gastric residue in mice, showing significant differences compared with the blank group (all p<0.01), indicating that the traditional Chinese medicine composition has the effect of promoting gastric emptying. The results are shown in the table below.

[0525] Table 17 Effects of Traditional Chinese Medicine Compositions on Gastric Emptying and Small Intestinal Propulsion in Mice

[0526]

[0527]

[0528] Compared with the blank group: * P<0.05,** P<0.01.

[0529] In summary, the herbal composition of this invention exhibits significant pharmacodynamic effects in treating chronic non-atrophic gastritis (by lowering gastric juice pH, increasing free gastric acidity, reducing pepsin activity, decreasing free gastric mucus content, lowering serum gastrin and IL-8 levels, increasing serum PGE2 levels, and improving gastric histopathological changes), inhibiting Helicobacter pylori (HP), providing analgesia, anti-inflammation, and promoting gastrointestinal motility. Among these, increasing serum PGE2 levels, downregulating serum IL-8 and gastrin levels, and inhibiting HP may be key mechanisms in the therapeutic effect of this drug on chronic non-atrophic gastritis complicated with Helicobacter pylori infection. These findings provide experimental evidence for the clinical treatment of chronic non-atrophic gastritis complicated with Helicobacter pylori infection.

[0530] Example 7: Methodological Investigation of Astragaloside A Content Determination by High Performance Liquid Chromatography

[0531] 1. Specificity Examination

[0532] ① Prepare granules according to the prescription and preparation method described in Example 1, and then prepare the test solution and the reference solution according to the preparation method of the test solution and the reference solution in "A. Determination of Astragaloside A content" in Example 8, respectively; ② Prepare granules without Astragalus according to the prescription ratio and preparation method described in Example 1, i.e., Astragalus negative reference, and then prepare the Astragalus negative reference solution according to the test solution preparation method described in Example 8.

[0533] Following the chromatographic conditions in Example 8, the test solution, reference solution, and Astragalus negative control solution were injected into the chromatographic column, respectively. The results are shown in [Figure 8]. Figure 6 As shown, the separation degree between the astragaloside A peak and other impurity peaks in the test solution is greater than 1.5, indicating good separation. Furthermore, the astragaloside negative control solution did not show any chromatographic peaks at the same retention time as astragaloside A, so it is considered to have no interference and strong specificity.

[0534] 2. Examination of linear relationships

[0535] Accurately pipette the above reference solution and dilute it to obtain a series of standard solutions. Accurately pipette 10 μl of each solution and inject it into the liquid chromatograph. Measure the peak area according to the chromatographic conditions in Example 8, "A. Determination of Astragaloside A Content". Plot a standard curve with the logarithm of the astragaloside A peak area (Y) as the ordinate and the logarithm of the injection volume (X) as the abscissa. Perform linear regression to obtain the regression equation Y = 1.612X + 4.852, r = 0.9990. Figure 9As shown. The experimental results indicate that the method described in Example 8, "A. Determination of Astragaloside A Content," can effectively detect astragaloside A. Astragaloside A exhibits good linearity in the range of 3.896–23.376 μg.

[0536] 3. Precision test

[0537] The above-mentioned reference solution was accurately measured and injected six times under the chromatographic conditions described in Example 8, "A. Determination of Astragaloside A Content". The peak area of ​​astragaloside A was measured, and the RSD value of the peak area was calculated. The experimental results showed that the RSD value of the peak area of ​​astragaloside A was 0.97%, indicating that the instrument precision under these chromatographic conditions was good.

[0538] 4 Repeatability Test

[0539] The granules from Example 1 were used as a reference. Six test solutions were prepared in parallel under the chromatographic conditions described in Example 8, "A. Determination of Astragaloside A Content". The peak area of ​​astragaloside A was measured, and the content and RSD value were calculated. The RSD value of astragaloside A content was 2.09%. The results showed that the method had good repeatability.

[0540] 5 Recovery rate test

[0541] The 100% spiking recovery method was used. Approximately 2.5 g of the granules from Example 1 was accurately weighed, divided into six portions. Six test solutions were prepared according to the method described in Example 8 for "A. Determination of Astragaloside A Content". A certain amount of reference solution was added to each solution, and the chromatographic conditions described in Example 1 were followed. The recovery rate was calculated using the following formula: Recovery rate = (Measured value - Test content × Sample amount) / Added amount × 100%. The average recovery rate of astragaloside A was 103.36%, with an RSD of 2.14%. The results indicate that the recovery rate of this method meets the requirements.

[0542] Example 7: Methodological Investigation of the Determination of Paeoniflorin and Naringin by High Performance Liquid Chromatography

[0543] 1. Specificity Examination

[0544] ① Prepare granules according to the prescription and preparation method described in Example 2, and then prepare the test solution and the reference solution according to the preparation method of the test solution and the reference solution in the method of "B. Determination of the content of paeoniflorin and naringin" in Example 8, respectively; ② Prepare granules without red peony root and bitter orange peel according to the prescription ratio and preparation method described in Example 2, that is, red peony root and bitter orange peel double negative preparation, and then prepare red peony root and bitter orange peel double negative reference solution according to the preparation method of the test solution in the method of "B. Determination of the content of paeoniflorin and naringin" in Example 8.

[0545] Following the chromatographic conditions in the "B. Determination of the content of paeoniflorin and naringin" method in Example 8, the test solution, reference solution, and double negative reference solutions of Paeonia lactiflora and Citrus aurantium were injected into the chromatographic column, respectively. The results are shown in the figure. Figures 7-8 As shown, the separation degree of paeoniflorin and naringin peaks from other impurity peaks in the test solution is greater than 1.5, indicating good separation. Furthermore, the double negative control solutions of Paeonia lactiflora and Citrus aurantium do not show chromatographic peaks at the same retention time as paeoniflorin and naringin, so they are considered to have no interference and strong specificity.

[0546] 2. Examination of linear relationships

[0547] Accurately pipette the paeoniflorin reference solution, dilute to obtain a series of standard solutions, and accurately inject 10 μl of each into the liquid chromatograph. Measure the peak area according to the paeoniflorin chromatographic conditions. Plot a standard curve with the peak area value of paeoniflorin (Y) as the ordinate and the injection volume (X) as the abscissa, obtaining the regression equation Y = 1323.5251X - 12.2812, r 2 =1, such as Figure 10 As shown in the figure. The results indicate that the detection method described in "B. Determination of the content of paeoniflorin and naringin" in Example 8 can effectively detect paeoniflorin, and paeoniflorin has a good linear relationship in the range of 0.5176 to 10.352 μg.

[0548] Accurately pipette the naringin reference solution, dilute to obtain a series of standard solutions, and accurately inject 10 μl of each into the liquid chromatograph. Measure the peak area according to the chromatographic conditions for naringin. Plot a standard curve with the peak area value of naringin (Y) as the ordinate and the injection volume (X) as the abscissa, obtaining the regression equation Y = 1601.5X + 0.045, r 2 =1, such as Figure 11 As shown. The results indicate that the detection method described in Example 8, "B. Determination of the content of paeoniflorin and naringin", can effectively detect naringin, and naringin has a good linear relationship in the range of 0.06716 to 1.3432 μg.

[0549] 3. Precision test

[0550] The precision of the instrument was assessed using paeoniflorin and naringin as indicators. The aforementioned reference solutions were precisely measured and injected six times under the chromatographic conditions described in Example 8, "B. Determination of the Content of Paeoniflorin and Naringin." The peak areas of paeoniflorin and naringin were measured, and the RSD values ​​of the peak areas were calculated. The results showed that the RSD value of paeoniflorin was 0.07%, and the RSD value of naringin was 0.10%, indicating good instrument precision under these chromatographic conditions.

[0551] 4 Repeatability Test

[0552] The above-mentioned granules were used as the test solution for six samples prepared in parallel with the method described in "B. Determination of the content of paeoniflorin and naringin" in Example 8. The peak areas of paeoniflorin and naringin were measured, and the contents and RSD values ​​were calculated. The results showed that the average content of paeoniflorin was 3.55 mg / g with an RSD value of 0.48%; the average content of naringin was 6.02 mg / g with an RSD value of 0.70%. The results indicate that the reproducibility of this method is good.

[0553] 5 Recovery rate test

[0554] The 100% spiking recovery method was used. Approximately 0.65 g of the above-mentioned granules were accurately weighed, divided into six portions. Six test solutions were prepared according to the method described in Example 8 for the "Determination of Contents of B, Paeoniflorin and Naringin". A certain amount of reference solution was added to each solution, and the chromatographic conditions were determined according to the description in Example 8 for the "Determination of Contents of B, Paeoniflorin and Naringin". The recovery rate was calculated using the following formula: Recovery rate = (Measured value - Test content × Sample amount) / Added amount × 100%. The average recovery rate of paeoniflorin was 101.17%, with an RSD of 2.10%; the average recovery rate of naringin was 98.33%, with an RSD of 0.66%. The results show that the recovery rate of this method meets the requirements.

[0555] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A traditional Chinese medicine composition for treating chronic gastritis, characterized in that, Made from the following parts by weight of raw materials: Astragalus membranaceus 15-25 parts by weight, dandelion 15-25 parts by weight, angelica sinensis 6-15 parts by weight, atractylodes macrocephala 10-20 parts by weight, turmeric 6-15 parts by weight, immature bitter orange 6-15 parts by weight, red peony root 10-20 parts by weight, coptis chinensis 2-8 parts by weight, cinnamon 2-8 parts by weight, licorice root 6-15 parts by weight.

2. The traditional Chinese medicine composition for treating chronic gastritis according to claim 1, characterized in that, Made from the following ingredients in parts by weight: Astragalus membranaceus 20 parts by weight, Taraxacum mongolicum 20 parts by weight, Angelica sinensis 10 parts by weight, Atractylodes macrocephala 15 parts by weight, Curcuma zedoaria 10 parts by weight, Citrus aurantium 10 parts by weight, Paeonia lactiflora 15 parts by weight, Coptis chinensis 5 parts by weight, Cinnamomum cassia 5 parts by weight, Glycyrrhiza uralensis 10 parts by weight; or Astragalus membranaceus 15 parts by weight, dandelion 15 parts by weight, angelica sinensis 6 parts by weight, atractylodes macrocephala 10 parts by weight, turmeric 6 parts by weight, immature bitter orange 6 parts by weight, red peony root 10 parts by weight, coptis chinensis 2 parts by weight, cinnamon 2 parts by weight, licorice root 6 parts by weight; or Astragalus membranaceus 18 parts by weight, dandelion 18 parts by weight, angelica sinensis 9 parts by weight, atractylodes macrocephala 14 parts by weight, turmeric 9 parts by weight, immature bitter orange 9 parts by weight, red peony root 14 parts by weight, coptis chinensis 4 parts by weight, cinnamon 4 parts by weight, licorice root 9 parts by weight; or Astragalus membranaceus 17 parts by weight, dandelion 17 parts by weight, angelica sinensis 8 parts by weight, atractylodes macrocephala 12 parts by weight, turmeric 8 parts by weight, immature bitter orange 8 parts by weight, red peony root 12 parts by weight, coptis chinensis 4 parts by weight, cinnamon 4 parts by weight, and licorice root 8 parts by weight.

3. A method for preparing a traditional Chinese medicine composition for treating chronic gastritis as described in claim 1 or 2, characterized in that, Includes the following steps: (1) Weigh out the volatile oils of Angelica sinensis, Atractylodes macrocephala, Curcuma zedoaria, Citrus aurantium and Cinnamomum cassia according to the selected weight proportions, add water and distill to extract the volatile oils, collect the volatile oils, and incorporate them to obtain the incorporation complex for later use; (2) Weigh out dandelion, red peony root and licorice root according to the selected weight proportions, extract with water, filter, and keep the filtrate for later use; (3) Weigh out Astragalus membranaceus and Coptis chinensis according to the selected weight proportions, extract with ethanol solution, filter, and keep the filtrate for later use; (4) Mix the filtrates obtained in steps (2) and (3) and dry them. Add the volatile oil inclusion complex and then add or not add conventional excipients to make a clinically acceptable dosage form.

4. The method for preparing the traditional Chinese medicine composition for treating chronic gastritis according to claim 3, characterized in that, In the inclusion step of step (1), the ratio of volatile oil, β-cyclodextrin and water is 1ml: 5-20g: 40-100ml; the inclusion time is 5-30 minutes; and the inclusion mixture is refrigerated for 12-48 hours after inclusion.

5. The method for preparing the traditional Chinese medicine composition for treating chronic gastritis according to claim 3 or 4, characterized in that, In step (1) of water distillation extraction, add 8-14 times the amount of water and steam distill for 4-8 hours; And / or, in the water extraction step (2), add water and decoct 1-3 times, 1-3 hours each time, adding 6-14 times the amount of water each time; And / or, in step (3) ethanol extraction, the extract is heated and refluxed with a 50 v / v%-80 v / v% ethanol solution for 1-3 times, each time for 1-2 hours, with 6-12 times the volume of the 50 v / v%-80 v / v% ethanol solution added each time. The extract is filtered and combined. And / or, the dosage form is a tablet, granule, capsule, oral liquid, or injection.

6. A method for detecting a traditional Chinese medicine composition for treating chronic gastritis as described in claim 1 or 2, or a traditional Chinese medicine composition for treating chronic gastritis prepared by the preparation method according to any one of claims 3-5, characterized in that, The following methods are included for content determination: A. Determination of astragaloside A content: Chromatographic conditions: Determined by high performance liquid chromatography (HPLC), using octadecylsilane-bonded silica gel as the packing material; acetonitrile as mobile phase A and water as mobile phase B, with a volume ratio of mobile phase A:mobile phase B of 20-40%:80-60%; drift tube temperature of the evaporative light scattering detector 35-100℃; gas pressure 300-400 kPa; theoretical plate number calculated based on astragaloside A should not be less than 5000. Determination method: Pipe the reference solution and the test solution separately into the liquid chromatograph, measure and calculate.

7. The detection method for the traditional Chinese medicine composition for treating chronic gastritis according to claim 6, characterized in that, The determination of the content of A and astragaloside A also includes: Preparation of the test solution: Take the Chinese herbal composition, add methanol solution, heat to reflux, cool, filter, evaporate the filtrate to dryness, dissolve the residue in water, extract with water-saturated n-butanol, combine the n-butanol solutions, wash, discard the washing liquid, evaporate to dryness, dissolve the residue in methanol and make up to volume to obtain the test solution. Preparation of reference solution: Take astragaloside A reference standard and add organic solvent to prepare reference solution.

8. The method for detecting the traditional Chinese medicine composition for treating chronic gastritis according to claim 7, characterized in that, The concentration of the methanol solution is 70-100 v / v%. And / or, the mass of the traditional Chinese medicine composition is 5g, and the amount of methanol solution used is 20-150ml; And / or, the heating reflux time is 0.1-1.0 h; And / or, extract with water-saturated n-butanol 1-5 times, 10-50 ml each time; And / or, dissolve the residue in methanol and bring the volume to 2-50 ml.

9. The method for detecting the traditional Chinese medicine composition for treating chronic gastritis according to claim 6, 7, or 8, characterized in that, It also includes at least one of the following content determination methods or identification methods: B. Determination of the content of paeoniflorin and naringin: Chromatographic conditions: The determination was performed using high performance liquid chromatography (HPLC), with octadecylsilane-bonded silica gel as the stationary phase; acetonitrile as mobile phase A and 0.05-0.4 v / v% phosphoric acid solution as mobile phase B, with gradient elution. The volume ratio of mobile phase A to mobile phase B was 5-80%:95-20%; the detection wavelength was 220-290 nm; the theoretical plate number, calculated based on naringin, should be no less than 5000. Determination method: Accurately pipette the reference solution and the test solution separately and inject them into the liquid chromatograph, measure and calculate; or C. Identification of Angelica sinensis: Preparation of the test solution: Take 0.5-4g of the traditional Chinese medicine composition, add 10-40ml of ether, sonicate for 5-30 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1-5ml of ethanol to obtain the test solution; Preparation of the reference herb solution: Take 0.1-2g of Angelica sinensis reference herb and prepare the reference herb solution according to the preparation method of the test sample solution; Identification: Perform thin-layer chromatography (TLC). Apply 2-10 μl each of the test solution and the reference medicinal material solution to the same silica gel G TLC plate. Develop the plate using a mixture of n-hexane and ethyl acetate in a volume ratio of 2-5:1-3. Remove the plate, air-dry, and examine. In the chromatogram of the test sample, fluorescent spots of the same color should appear at the corresponding positions as in the chromatogram of the reference medicinal material. D. Identification of Atractylodes macrocephala: Preparation of the test solution: Take 0.5-4g of the traditional Chinese medicine composition, add 5-30ml of n-hexane, sonicate for 10-60 minutes, cool, filter, and take the filtrate to obtain the test solution; Preparation of the reference herb solution: Take 0.5-2g of Atractylodes macrocephala reference herb and prepare the reference herb solution according to the preparation method of the test sample; Identification: Perform thin-layer chromatography (TLC). Apply 1-10 μl each of the test solution and the reference medicinal material solution to the same silica gel G TLC plate. Develop the plate using a mixture of petroleum ether and ethyl acetate at a volume ratio of 20-100:1-5 (60-90℃). Remove the plate, air dry, spray with 1-10% w / v vanillin-sulfuric acid solution, and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material; or... E. Identification of Citrus aurantium: Preparation of the test solution: Take 0.5-4g of the Chinese herbal composition, add 5-30ml of 40-100v / v% methanol, sonicate for 10-60 minutes, cool, filter, evaporate the filtrate to dryness, add 1-5ml of methanol to dissolve the residue to obtain the test solution. Preparation of reference solutions: Take naringin reference standard and neohesperidin reference standard, add 40-100 v / v % methanol to prepare solutions containing 0.2-2 mg per 1 ml, respectively, as reference solutions; Identification: Perform thin-layer chromatography (TLC). Apply 1-10 μl each of the test solution and the reference solution to the same silica gel G TLC plate. Use the lower layer of a mixture of chloroform, methanol, and water (volume ratio 10-15:2-10:1-4) as the developing solvent. Develop to a distance of 8-20 cm. Remove, air dry, spray with 1-5% w / v aluminum chloride ethanol solution, heat at 100-110℃ for approximately 3-6 minutes, and examine under ultraviolet light. The test sample chromatogram should show fluorescent spots of the same color at the corresponding positions as the reference sample chromatogram; or... F. Identification of Paeonia lactiflora: Preparation of the test solution: Take 0.5-4g of the Chinese herbal composition, add 5-30ml of 40-100% ethanol, shake for 5-30 minutes, filter, evaporate the filtrate to dryness, and dissolve the residue in 1-5ml of 40-100% ethanol to obtain the test solution. Preparation of reference solution: Take paeoniflorin reference standard and add 40-100 v / v % methanol to prepare a solution containing 0.2-2 mg per 1 ml, which is used as the reference solution; Identification: Perform thin-layer chromatography. Apply 1-10 μl each of the test solution and the reference solution to the same silica gel G thin-layer plate. Use a mixture of chloroform-ethyl acetate-methanol-formic acid (v / v) at a ratio of 20-50:2-10:5-15:0.1-1 as the developing solvent. Develop, remove, and air-dry. Spray with 5% vanillin-sulfuric acid-ethanol solution and heat until the spots are clearly visible. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference sample; or... G. Identification of Coptis chinensis: Preparation of the test solution: Take 0.5-4g of the traditional Chinese medicine composition, add 5-30ml of 40-100 v / v % methanol, sonicate for 5-30 minutes, filter, and take the filtrate to obtain the test solution; Preparation of reference solution: Take berberine hydrochloride reference standard and add 40-100 v / v % methanol to prepare a solution containing 0.2-2 mg per 1 ml, which is used as the reference solution; Identification: Perform thin-layer chromatography. Take 1-10 μl each of the above-mentioned test solution and reference solution and spot them separately on the same silica gel G thin-layer plate. Use a solution of cyclohexane, ethyl acetate, isopropanol, methanol, water, and triethylamine in a volume ratio of 1-5:1-5:0.5-3:0.5-3:0.5-3:0.5-3 as the developing solvent. Place the plate in a developing tank pre-saturated with concentrated ammonia solution for 20 minutes, develop, remove, air dry, and examine under ultraviolet light. In the chromatogram of the test sample, fluorescent spots of the same color appear at the corresponding positions as in the chromatogram of the reference sample.

10. The method for detecting the traditional Chinese medicine composition for treating chronic gastritis according to claim 9, characterized in that, In the determination of the contents of B, paeoniflorin and naringin: The gradient elution procedure is as follows: 0~9 min, 15 v / v %A; 9~30 min, 15 v / v %A → 22 v / v %A; 30~33 min, 22 v / v %A → 50 v / v %A; 33~35 min, 50 v / v %A; And / or, the detection wavelengths are 230nm and 283nm; And / or, mobile phase B is a 0.1 v / v% phosphoric acid solution; And / or, preparation of the test solution: Take 0.5-3g of the traditional Chinese medicine composition, add 10-100mL of methanol with a volume concentration of 40-100v / v%, seal tightly, weigh, reflux in a water bath for 1-3 hours, cool, weigh again, replenish the lost weight with 40-100% methanol, shake well, filter, and take the filtrate to obtain the test solution; And / or, preparation of reference solutions: Take appropriate amounts of paeoniflorin and naringin reference standards, accurately weigh them, and add 50-100 v / v% methanol solution to prepare paeoniflorin and naringin reference solutions. And / or, the injection volume is 2-20 μL.

11. The traditional Chinese medicine composition for treating chronic gastritis according to claim 1 or 2, or the traditional Chinese medicine composition for treating chronic gastritis prepared by the preparation method according to any one of claims 3-5, has any of the following uses: (1) Use in the preparation of drugs for treating chronic non-atrophic gastritis complicated with Helicobacter pylori infection; (2) Use in the preparation of drugs for treating Helicobacter pylori infection.

12. The use according to claim 11, characterized in that, The medication is used to treat bloating, stomach pain, acid reflux, and heartburn.

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