Production method of traditional Chinese medicine composition based on thin layer identification method

Abstract

The invention provides a preparation method of a traditional Chinese medicine composition based on a thin-layer chromatography identification method. The method comprises the following steps: weighing the following traditional Chinese medicine components in parts by weight: 19-21 parts of old forsythia, 14-16 parts of ginger pinellia, 11-13 parts of roasted liquorice, 11-13 parts of bupleurum, 9-11 parts of codonopsis, 7-9 parts of dried ginger, 5-7 parts of scutellaria, 4-6 parts of yalian, 5-7 parts of cooked rhubarb, and 18-22 parts of jujube; after preparing the traditional Chinese medicine composition, selecting forsythia, coptis, and liquorice as quality active ingredients for quality control. Among them, for Forsythia suspensa, chloroform and methanol are mixed in a volume ratio of 8:1 as a developing agent; for Coptis chinensis, toluene, ethyl acetate, methanol, isopropanol and water are mixed in a volume ratio of 6:3:2:1.5:0.3 as a developing agent, and placed in a developing cylinder pre-saturated with concentrated ammonia test solution for 20 minutes; for Licorice, ethyl acetate, glacial acetic acid and water are mixed in a volume ratio of 15:2:2 as a developing agent, and quality monitoring is carried out, and standardized production of products is guided based on the quality monitoring results.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine production methods, and particularly relates to a production method of a traditional Chinese medicine composition based on a thin layer identification method. Background Art

[0002] Existing medications for treating oral ulcers are primarily Western medicines, including antiseptic and anti-inflammatory drugs such as chlorhexidine gargle, cediridine lozenges, and dequalinium chloride lozenges; analgesics such as compound chamomile lidocaine gel and compound benzocaine gel; and vitamins such as vitamin C and vitamin B complex. While these medications have some efficacy, they are associated with significant side effects and are prone to recurrence after discontinuation. There are also some traditional Chinese medicine preparations, such as Yangyin Shengji Powder, Bingpeng Powder, and Xiguashuang. These medications are effective for occasional oral ulcers, primarily to alleviate pain or reduce recurrences, but they are not effective for treating stubborn or recurrent oral ulcers.

[0003] Chinese invention patent application 201910134460.6, "A Traditional Chinese Medicine Composition for Treating Oral Ulcers," discloses a traditional Chinese medicine composition for treating oral ulcers, comprising scutellaria baicalensis, coptis chinensis, pinellia ternata, codonopsis pilosula, angelica dahurica, raw cimicifuga heracleifolia, dandelion, radish seed, roasted ginger, patchouli, calamus tatarinowii, gardenia seed, gypsum, bupleurum chinense, platycodon grandiflorum, astragalus root, forsythia suspensa, capillaris scabra, mulberry bark, and raw licorice. These ingredients work together to clear heat, detoxify, and relieve pain. In this invention, scutellaria baicalensis, coptis chinensis, pinellia ternata, bupleurum root, and raw cimicifuga are the primary ingredients, providing heat-clearing, detoxifying, and relieving pain. Astragalus membranaceus has the effects of invigorating qi and strengthening the exterior, arresting perspiration and preventing eczema, promoting wound healing and promoting tissue regeneration, and promoting diuresis and reducing swelling; Coptis chinensis has the effects of clearing heat and dampness, purging fire and detoxifying; Pinellia ternata has the effects of drying dampness and resolving phlegm, dissolving lumps and dispersing stagnation; Bupleurum chinense has the effects of harmonizing the exterior and interior, soothing the liver and raising yang; and Cimicifuga heracleifolia has the effects of relieving rashes, clearing heat and detoxifying, and raising yang and lifting sunken areas. In the present invention, Astragalus membranaceus, Codonopsis pilosula, and Licorice root are combined to invigorate qi and strengthen the spleen. Modern medical research indicates that Astragalus membranaceus, Codonopsis pilosula, and Licorice root can regulate the body's immunity and enhance disease resistance. Morus alba bark is combined with Scutellaria baicalensis to clear heat and eliminate dampness. Platycodon grandiflorum and Licorice root, combined with Astragalus membranaceus, Codonopsis pilosula, and Licorice root, combine to address both deficiency and excess, clearing heat and eliminating dampness without harming the body. Cimicifuga heracleifolia and Bupleurum chinense raise clear qi, and when combined with Gypsum sutsuga, Artemisia capillaris, and Angelica dahurica, they enhance the effect of clearing fire in the Yangming Stomach Meridian; Bupleurum chinense soothes the liver and relieves depression. Licorice and Platycodon grandiflorum can be used together to clear away heat, detoxify and relieve pain, and can also carry medicine to the oral cavity. Forsythia suspensa is a "holy medicine for sores". When used with dandelion, it can enhance the ability to clear away the heat and evil of the exterior and interior. Although Paojiang is a hot medicine that enhances the effect of warming the middle and dispersing cold, it can also control the cold nature of heat-clearing and detoxifying medicines, expelling evil without going up to the right path. It is more effective for those with recurrent mouth sores that do not heal for a long time. Patchouli and Acorus gramineus can clear away damp-heat in the middle jiao spleen and stomach, while radish seeds can promote the flow of qi. When the qi is in harmony, the disease will heal itself. This invention uses Scutellaria baicalensis, Coptis chinensis, Pinellia ternata, Bupleurum chinense and Cimicifuga heracleifolia as the main medicines (monarch medicines), which have the effect of clearing away heat and detoxifying, purging fire and relieving pain. Other medicinal flavors assist the monarch medicine in clearing away heat and detoxifying, purging fire and relieving pain by improving immunity, clearing heat and removing dampness.

[0004] Chinese invention patent application 201711037406.7 "An oral medicine for treating recurrent oral ulcers and its preparation method" discloses an oral medicine for treating recurrent oral ulcers. The components of the medicine and their mass proportions are as follows: 3-6 parts of coptis chinensis, 15-65 parts of gypsum, 10-16 parts of redhead grass, 3-10 parts of scutellaria baicalensis, 6-15 parts of olive, 8-16 parts of kudzu root, 15-35 parts of radix scutellariae, 3-12 parts of immature bitter orange, 10-16 parts of rose root, 3-12 parts of liquorice, 6-10 parts of smilax china, 3-9 parts of pinellia, 12-35 parts of rehmannia root, 15-28 parts of panax quinquefolium, 6-16 parts of white peony root, and 3-12 parts of bupleurum. The invention uses coptis root, scutellaria baicalensis, gypsum and redhead grass which have the effects of clearing heat, detoxifying and purging fire as the main medicines, and uses olive, kudzu root, immature bitter orange and other auxiliary medicines or adjuvants for conditioning.

[0005] Chinese invention patent application 201210173106.2, "A Traditional Chinese Medicine Composition for Treating Oral Ulcers," discloses a traditional Chinese medicine composition for treating oral ulcers. The composition comprises, by weight, 13 parts honeysuckle, 13 parts forsythia, 15 parts coptis, 15 parts schisandra, 12 parts costus root, 12 parts poria, 10 parts raw licorice, and 10 parts magnetite. This invention utilizes the heat-clearing and detoxifying properties of honeysuckle and coptis, as well as the wind-heat dispersing properties, supplemented by forsythia's detoxifying and antidiarrheal properties. The composition is then formulated with adjuvants such as licorice, schisandra, and magnetite to create a powder for external application to the affected area, achieving excellent results.

[0006] Chinese invention patent application 201210173106.2, entitled "A Traditional Chinese Medicine Composition for Treating Oral Ulcers," discloses a traditional Chinese medicine composition for treating oral ulcers. The composition comprises, by weight, 13 parts honeysuckle, 13 parts forsythia, 15 parts coptis, 15 parts schisandra, 12 parts costus root, 12 parts poria, 10 parts raw licorice, and 10 parts magnetite. This invention utilizes the heat-clearing and detoxifying properties of honeysuckle and coptis, as well as the wind-heat dispersing properties, supplemented by forsythia's detoxifying and antidiarrheal properties. The composition is then formulated with adjuvants such as licorice, schisandra, and magnetite to create a powder for external application to the affected area, achieving excellent results.

[0007] Chinese invention patent application 201810375215.X, titled "An Oral Spray and Its Preparation Method," discloses an oral spray for treating oral ulcers. The spray comprises, by weight, 3-6 parts of propolis total flavonoids, 20-25 parts of honeysuckle extract, 30-40 parts of forsythia extract, 5-10 parts of menthol, and 15-20 parts of borneol, as well as 5-10 parts of alcohol, 3-5 parts of propylene glycol, and 10-20 parts of polyethylene glycol with a molecular weight of no more than 600. While the spray can achieve good results in the short term, it is a topical formulation. According to traditional Chinese medicine theory, topical medications only temporarily relieve superficial symptoms without treating the corresponding internal symptoms or regulating the patient's immune system. Therefore, the condition is prone to recurrence over time.

[0008] Furthermore, Traditional Chinese Medicine (TCM) theory holds that while oral ulcers manifest in the mouth, they are primarily caused by visceral dysfunction leading to disharmony between the spleen and stomach. This leads to excessive internal heat in the heart, liver, and stomach, which in turn causes a buildup of toxins that cannot be effectively metabolized and then circulate in the mouth, causing inflammation of the internal heat. Recurrent oral ulcers, if not effectively treated and controlled, can develop into Behçet's syndrome, blindness, or even malignant transformation. Studies have shown that conditions such as gastric ulcers, duodenal ulcers, atrophic gastritis, and other gastrointestinal disorders can lead to endocrine disorders, which can contribute to the development of oral ulcers. Consequently, oral ulcers are sometimes associated with acute and chronic gastroenteritis and Behçet's syndrome.

[0009] However, existing traditional Chinese medicine compositions for treating acute and chronic gastroenteritis and fox-induced madness (Behcet's syndrome) are mostly based on Gancao Xiexin Decoction or Lizhong Decoction, which are obtained by adjusting and adding or subtracting the formula. Although they have certain efficacy for acute gastroenteritis, they are not very effective for oral ulcers caused by diseases such as acute gastroenteritis.

[0010] Traditional Chinese medicine theory holds that while oral ulcer symptoms manifest in the mouth, they are primarily caused by spleen and stomach weakness and gastrointestinal dysfunction, leading to the upward flow of heart, liver, and stomach fire into the mouth. Therefore, existing Chinese herbal compositions, which only clear heat and detoxify, purge fire, and relieve pain, can temporarily alleviate clinical symptoms but are unable to effectively treat gastrointestinal dysfunction, making it difficult to achieve a complete cure for oral ulcers.

[0011] In summary, the above existing invention patents all use the basic treatment principles of clearing away heat and detoxifying, purging fire and relieving pain for the treatment of oral ulcers, supplemented by formulas of related auxiliary and adjuvant drugs. They are basically consistent with the treatment principles of existing marketed Chinese patent medicines (such as Yangyin Shengji Powder, Bingpeng Powder, Xigua Shuang, etc.). Based on the above treatment principles, good clinical results can be achieved in a short period of time, temporarily alleviating the "symptoms". However, since the "root cause" is not thoroughly treated, it is still prone to recurrence over a long period of time.

[0012] Therefore, the prior art requires a Chinese medicine composition that can treat both the symptoms and the root cause of oral ulcers. The composition should be able to quickly relieve the pain of oral ulcers and prevent the recurrence of oral ulcers, and have the effects of simple prescriptions, easy standardization and industrialization.

[0013] In addition, a method for quickly and easily monitoring the product quality of the traditional Chinese medicine composition is also needed, as well as a standardized production method based on the quality monitoring method. Summary of the Invention

[0014] The first object of the present invention is to provide a traditional Chinese medicine composition for treating oral ulcers, which comprises the following components in parts by weight:

[0015] 19-21 parts of Forsythia suspensa; 14-16 parts of Pinellia ternata; 11-13 parts of Radix Glycyrrhizae; 11-13 parts of Radix Bupleuri; 9-11 parts of Codonopsis pilosula; 7-9 parts of Dry Ginger; 5-7 parts of Scutellaria baicalensis; 4-6 parts of Coptis chinensis; 5-7 parts of Rhubarb; 18-22 parts of Jujube.

[0016] In one embodiment: 20 parts of Forsythia suspensa, 15 parts of Pinellia ternata, 12 parts of Radix Glycyrrhizae Preparata, 12 parts of Radix Bupleuri, 10 parts of Codonopsis pilosula, 8 parts of Dry Ginger, 6 parts of Scutellaria baicalensis, 5 parts of Coptis chinensis, 6 parts of Rhubarb Rhizome, and 20 parts of

[0017] The traditional Chinese medicine composition further comprises the components in the above-mentioned weight ratio and pharmaceutically acceptable excipients.

[0018] The Chinese medicine composition is an oral preparation, preferably granules, tablets, capsules, pills, powders, etc.

[0019] In another preferred embodiment, the excipients of the formulation are disintegrants, fillers, binders, lubricants and / or bases.

[0020] Furthermore, the disintegrant includes one or more of starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinyl pyrrolidone, low-substituted hydroxypropyl cellulose, and cross-linked sodium carboxymethyl cellulose;

[0021] Furthermore, the filler includes one or more of starch, mannitol, chitin, pregelatinized starch, lactose, microcrystalline cellulose, and sucrose;

[0022] Furthermore, the lubricant includes one or more of sodium lauryl sulfate, magnesium stearate, talc, and silicon dioxide;

[0023] Furthermore, the adhesive comprises one or more of starch slurry, hydroxypropyl methylcellulose and polyvinyl pyrrolidone;

[0024] Furthermore, the matrix includes one of maltodextrin or PEG4000 (polyethylene glycol 4000) and PEG6000 (polyethylene glycol 6000);

[0025] In any of the above embodiments, the Chinese medicine composition can be used to treat oral ulcers, acute and chronic gastroenteritis and Behcet's syndrome.

[0026] In the present invention, the pharmacological effects of the Chinese herbal medicine raw materials used are as follows:

[0027] Forsythia suspensa (preferably aged): Cool in nature and bitter in flavor, it enters the Heart, Liver, and Gallbladder meridians. Indications: Clears heat and detoxifies, reduces swelling and resolves stagnation, and dispels wind-heat. It is used for throat infections, swollen tonsils, oral ulcers, carbuncles, scrofula, mastitis, wind-heat colds, the initial onset of febrile disease, invasion of warm and hot blood into the Ying meridians, high fever, polydipsia, coma, rash, and urinary retention due to heat.

[0028] Pinellia ternata: Pungent, warm, and toxic. It enters the spleen, stomach, and lung meridians. Functions and indications: It dries dampness and resolves phlegm, relieves adverse reactions, and stops vomiting. It is used for vomiting caused by phlegm and fluid retention, and for abdominal distension and fullness.

[0029] Roasted Licorice Root: Sweet and neutral in flavor. It nourishes the spleen and stomach, replenishes Qi and restores pulse. It is used for spleen and stomach deficiency, fatigue, palpitations, and irregular pulse.

[0030] Bupleurum: Bitter in nature and slightly cold. Functions and indications: Harmonizes the exterior and interior, soothes the liver, and promotes yang. Used for colds, fever, alternating chills and fever, chest and flank pain, irregular menstruation, uterine prolapse, and rectal prolapse.

[0031] Codonopsis pilosula: Sweet, slightly bitter, and slightly warm in nature. It enters the spleen, lung, heart, and kidney meridians. Benefits: Boosts vital energy, restores pulse and strengthens weakness, tonifies the spleen and lungs, promotes fluid production and nourishes blood, calms the mind and improves intelligence. It is used for symptoms of physical weakness, cold limbs and weak pulse, spleen deficiency, poor appetite, lung deficiency, wheezing and coughing, thirst due to loss of fluid, internal heat and polydipsia, qi and blood deficiency, chronic illness, palpitations and insomnia, impotence, and a cold uterus.

[0032] Dried ginger: Pungent and warming in nature. It enters the spleen, stomach, heart, and lung meridians. It warms the middle and dispels cold, restores yang and unblocks the meridians, and warms the lungs and eliminates fluid retention. It is primarily used to treat cold pain in the abdomen, vomiting and diarrhea, yang deficiency, cold extremities and weak pulse, and cough and asthma caused by phlegm and fluid retention.

[0033] Coptis chinensis (preferably Coptis chinensis): Cold in nature, bitter in flavor. It enters the Heart, Spleen, Stomach, Liver, Gallbladder, and Large Intestine meridians. It clears heat and dampness, purging fire and detoxifying. It belongs to the heat-clearing and dampness-relieving category of antipyretics. It is used to treat damp-heat-induced fullness, vomiting and acid regurgitation, diarrhea, jaundice, high fever and coma, hyperactive heart fire, vomiting and epistaxis due to blood heat, conjunctivitis, toothache, diabetes, carbuncles, and furuncles. Appropriate topical application can treat eczema, eczema, and abscesses.

[0034] Scutellaria baicalensis (preferably Scutellaria baicalensis): Bitter, cold. It enters the Lung, Gallbladder, Spleen, Large Intestine, and Small Intestine meridians. It clears lung heat from the upper burner, purging fire and detoxifying. It is used for lung-heat-induced cough, yellow phlegm, chest tightness and nausea, damp-heat fullness and distension, diarrhea, jaundice, high fever and thirst, vomiting and epistaxis due to blood heat, carbuncles, sores, and fetal restlessness.

[0035] Cooked Rhubarb: Bitter and cold in nature. It enters the spleen, stomach, large intestine, liver, and pericardium meridians. It has the effects of purging, clearing away heat and purging fire, cooling the blood and detoxifying, removing blood stasis and promoting menstruation, and promoting the elimination of dampness and jaundice. It is used for constipation due to excess heat, vomiting and bleeding due to blood heat, red eyes and swollen throat, carbuncles and furuncles, abdominal pain due to intestinal carbuncles, amenorrhea due to blood stasis, postpartum stasis, traumatic injuries, damp-heat dysentery, jaundice and dark urine, stranguria, and edema; it can also be used externally for burns and scalds.

[0036] Jujube: Sweet and warm in nature. It enters the spleen and stomach meridians. It tonifies the middle qi, nourishes the blood, calms the mind, and moderates the effects of other herbs. Indications: Used for spleen and stomach deficiency, poor appetite, loose stools, blood deficiency, and sallow complexion, as well as restlessness in women.

[0037] The first inventive principle of the present invention is that during the research on traditional Chinese medicine prescriptions for acute and chronic gastroenteritis and fox-induced madness (Behcet's syndrome), the inventors combined the Gancao Xiexin Decoction and Lizhong Decoction with Xiaochaihu Decoction and Sanhuang Xiexin Decoction according to the treatment principles. They unexpectedly found that the basic combination had a good therapeutic effect on oral ulcers and also retained a certain therapeutic effect on acute and chronic gastroenteritis. On the basis of the combined prescription, the inventors repeatedly optimized and adjusted it and finally simplified it into a simple prescription with 10 ingredients. This simple prescription, while retaining the main ingredients (Zingiber officinale, Licorice, and Bupleurum), added the main ingredient (Forsythia suspensa) to highlight the effects of clearing heat and detoxifying, dispersing nodules and eliminating carbuncles. At the same time, the dosage of Scutellaria baicalensis and Coptis chinensis, which are used to clear heat and detoxify and dissipate wind-heat, was reduced. The pharmacology was then adjusted by using dried ginger, jujube, Codonopsis pilosula, and cooked rhubarb. Finally, a traditional Chinese medicine composition was obtained that had a significant therapeutic effect on oral ulcers and also had a certain therapeutic effect on acute and chronic gastroenteritis and fox-induced madness (Behcet's syndrome).

[0038] Xiao Chaihu Tang, a traditional Chinese medicine formula, is a harmonizing agent with the effect of harmonizing the Shaoyang meridian. The formula, by weight, includes: 30 parts of Bupleurum, 9 parts each of Scutellaria, Ginseng, Pinellia, Roasted Licorice, and Sliced ​​Ginger, and 4 split jujubes. Bupleurum, a bitter and mild herb, enters the liver and gallbladder meridians, dispelling pathogenic heat and promoting the flow of qi through the meridians; Scutellaria clears away pathogenic heat; Pinellia soothes the stomach and reduces adverse effects; Ginseng and Roasted Licorice support vital energy and resist pathogenic factors; Ginger and jujube soothe stomach qi and promote body fluid production. This formula can dispel pathogenic qi, harmonize the Shaoyang meridian, clear the upper burner, promote the downward flow of body fluids, and harmonize stomach qi, resulting in sweating and fever relief. It is primarily used to treat Shaoyang syndrome caused by typhoid fever. The pathogen is located half-exterior and half-interior, with symptoms of alternating chills and fever, bitter fullness in the chest and flanks, silence, loss of appetite, restlessness and nausea, bitter taste in the mouth, dry throat, dizziness, a thin white tongue coating, and a wiry pulse. It also occurs in women with typhoid fever, with heat entering the blood chamber, causing menstruation to stop, and sporadic episodes of chills and fever. It also occurs in patients with internal injuries such as malaria and jaundice who exhibit the above Shaoyang symptoms. It is commonly used clinically to treat colds, influenza, malaria, chronic hepatitis, cirrhosis, acute and chronic cholecystitis, gallstones, acute pancreatitis, pleurisy, and otitis media, all of which are symptoms of disharmony between the gallbladder and stomach.

[0039] Sanhuang Xiexin Decoction originated in the 206 AD edition of the Golden Chamber. Its basic formula consists of rhubarb, coptis chinensis, and scutellaria baicalensis. It is a decoction with the effects of purging heat and detoxifying, and relieving dampness and heat. Adjustments to this basic formula include ingredients such as white schizonepeta, gardenia, arborvitae charcoal, madder, agrimony, peony bark, sanguisorba officinalis charcoal, calliandra, calcined cortex calophylla, radix trichosanthis, or liquorice. Clinically, it can treat upper gastrointestinal bleeding, acute pulmonary hemorrhage, and acne.

[0040] Licorice Decoction for Purging the Heart is a Chinese medicine prescription. It comes from the Treatise on Febrile Diseases. The prescription comprises, by weight, 12g of roasted licorice, 9g of scutellaria, 9g of dried ginger, 9g of pinellia, 12 jujubes, and 3g of coptis. This recipe is made by increasing the dosage of licorice in Banxia Xiexin Decoction. Licorice is the main ingredient, tonifying the middle and relieving urgency, replenishing a weak stomach and relieving acute diarrhea. The remaining ingredients also harmonize the stomach and eliminate bloating. It has the effects of invigorating qi and harmonizing the stomach, eliminating bloating and stopping vomiting. It is mainly used to treat bloating symptoms of typhoid fever, weak stomach qi, rumbling in the abdomen, diarrhea, indigestion, a hard and full lump under the heart, dry retching, restlessness, and fox-induced madness. It is commonly used clinically for acute and chronic gastrointestinal inflammation and Behçet's syndrome.

[0041] Lizhong Decoction, a traditional Chinese medicine formula, has multiple clinical formulations. The main one is based on ginseng, white atractylodes, roasted licorice root, and dried ginger, with the addition of bitter orange peel, poria, tangerine peel, or cinnamon twig. It is used to treat symptoms of spleen and stomach deficiency and cold, including spontaneous diarrhea without thirst, vomiting and abdominal pain, abdominal distension and loss of appetite, as well as cholera caused by cold, yang deficiency and blood loss such as hematemesis, bloody stool, or metrorrhagia, chest fullness and deficiency, pain extending to the back, fatigue, shortness of breath, and cold limbs. It is clinically used for patients with spleen and stomach deficiency and cold, such as acute and chronic gastritis, antral gastritis, ulcer disease, gastroptosis, and chronic hepatitis.

[0042] After the four prescriptions are combined, the medicinal ingredients include: Bupleurum, Scutellaria, Ginseng, Pinellia, Licorice (roasted), Jujube, Rhubarb, Coptis, Scutellaria, Licorice (roasted), Atractylodes, Dried Ginger, Citrus aurantium, Poria, Citrus reticulata, and Tangerine peel. At the same time, Forsythia suspensa is added, which can be used in combination with Scutellaria baicalensis and Coptis chinensis. While reducing the dosage of Scutellaria baicalensis and Coptis chinensis, the effects of Forsythia suspensa in clearing away heat, detoxifying, dispersing nodules and eliminating carbuncles are highlighted.

[0043] On this basis, in order to simplify the combination, Atractylodes macrocephala, Citrus aurantium, Poria cocos, Citrus aurantium peel, and Tangerine peel were discarded, and the content of components such as roasted licorice was adjusted to retain the efficacy of the above components;

[0044] Although the initial combined prescription contains ginseng, and both ginseng and Codonopsis have the effects of tonifying spleen qi, replenishing qi and promoting the production of body fluid, replenishing lung qi, replenishing qi and promoting the production of blood, and strengthening the body and dispelling pathogenic factors, they can be used for the treatment of spleen and lung qi deficiency and thirst caused by fluid deficiency in the late stage of febrile diseases, quenching thirst, blood deficiency, and symptoms of qi deficiency and pathogenic factors excess. However, Codonopsis has a sweet and mild nature and a milder effect, and can be used for treating mild and chronic diseases of the above symptoms. Therefore, the present invention uses Codonopsis to replace ginseng, and does not adopt ginseng suitable for acute and severe diseases.

[0045] The second object of the present invention is to provide a method for preparing the above-mentioned pharmaceutical composition, which is prepared from the following raw materials in the following weight ratio: 19-21 parts of Forsythia suspensa; 14-16 parts of Pinellia ternata; 11-13 parts of Radix Glycyrrhizae; 11-13 parts of Radix Bupleuri; 9-11 parts of Codonopsis pilosula; 7-9 parts of dried ginger; 5-7 parts of Scutellaria baicalensis; 4-6 parts of Coptis chinensis; 5-7 parts of Rhubarb; and 18-22 parts of jujube. The preparation method comprises the following steps:

[0046] (1) Decoction the above raw materials with water twice, each time for 0.5-1.0 hours, filter, and combine the decoctions;

[0047] (2) Concentrating the filtrate under reduced pressure at 60°C to a clear paste with a relative density of 1.03 to 1.10;

[0048] (3) adding an appropriate amount of matrix to the clear paste obtained in step (2), stirring evenly, and drying and granulating to obtain a clear paste powder;

[0049] (4) Monitor the quality of the dried paste powder and guide standardized production of the product based on the quality monitoring results;

[0050] (5) Based on the quality control results, different disintegrants, fillers, binders, lubricants and / or bases are added to the dry extract powder that meets the quality standards to prepare granules, tablets, capsules, pills or powders;

[0051] Furthermore, in step (2), 8-10 times the amount of water is added for the first time, soaked for 0.5 hours, and boiled for 1.0 hours; 6-8 times the amount of water is added for the second time, boiled for 0.5 hours, the decoctions are combined, and filtered;

[0052] Furthermore, in step (3), the filtrate obtained in step (2) is concentrated under reduced pressure at a temperature of 60°C ± 5°C to obtain a clear paste with a density of 1.03 to 1.05, and the clear paste is centrifuged at 10,000 rpm / min;

[0053] Furthermore, in step (4), the above-mentioned centrifuged clear paste is taken, an appropriate amount of maltodextrin is added, dissolved, mixed, and spray-dried to obtain a dry extract powder;

[0054] Furthermore, the quality monitoring of step (5) includes testing and evaluating the quality of the product and judging whether the product quality is qualified, thereby determining whether to continue or interrupt the production process of the product and guiding the standardized production of the product. In a specific embodiment, granulated samples are randomly sampled and thin layer chromatography identification is performed using the aforementioned developer group to determine whether to continue or interrupt the production process of the product.

[0055] The second principle of the present invention is to boldly explore and try various traditional Chinese medicine ingredients of the present invention, abandon the quality monitoring of some ingredients that are not necessary or have little impact on product quality, and conduct combination tests on various developing agents for major ingredients such as Forsythia suspensa, Licorice, and Coptis chinensis, so as to achieve a small number of identification developing agent groups to simultaneously detect multiple traditional Chinese medicine ingredients, thereby optimizing and simplifying the quality monitoring process while ensuring the identification effect.

[0056] The third object of the present invention is to provide a method for quality testing or monitoring the above-mentioned Chinese medicine composition, comprising the steps of:

[0057] (1) taking the dried paste powder, dissolving it in water, adding an extractant to extract it, and combining the extracts;

[0058] (2) Evaporate the extract to dryness in a water bath, and dissolve the residue in methanol as the test solution; prepare positive and negative control solutions in parallel using the same method;

[0059] (3) Pipette the test solution, positive control solution, and negative control solution separately, place them on the same silica gel G thin layer plate, develop them on the thin layer identification developer, and take them out;

[0060] (4) Add color developing solution, heat with hot air until the spots are clearly colored, and inspect under ultraviolet light;

[0061] (5) Compare the spots of the test sample, positive control, and negative control. If the test sample and the positive control have spots of the same color at the corresponding positions, and there are no interfering spots on the negative control, then the test sample is judged to contain the drug component of the same or similar quality as the positive control;

[0062] For Forsythia suspensa, chloroform and methanol were mixed in a volume ratio of 8:1 as the developing solvent;

[0063] For Coptis chinensis, mix toluene: ethyl acetate: methanol: isopropanol: water in a volume ratio of 6:3:2:1.5:0.3 as a developing agent and place it in a developing cylinder that has been pre-saturated with concentrated ammonia solution for 20 minutes;

[0064] For licorice, ethyl acetate: glacial acetic acid: water was mixed in a volume ratio of 15:2:2 as the developing solvent;

[0065] The traditional Chinese medicine composition comprises the following components in parts by weight: 19-21 parts of forsythia suspensa, 14-16 parts of pinellia ternata, 11-13 parts of roasted licorice, 11-13 parts of bupleurum, 9-11 parts of codonopsis pilosula, 7-9 parts of dried ginger, 5-7 parts of scutellaria baicalensis, 4-6 parts of coptis chinensis, 5-7 parts of cooked rhubarb, and 18-22 parts of jujube, and can be used to treat oral ulcers, acute and chronic gastroenteritis, and fox-induced madness.

[0066] In one embodiment, for Forsythia suspensa, steps (1) and (2) are extracted twice with n-butanol, the extracts are combined, evaporated in a water bath, and the residue is dissolved in methanol as the test solution; in another embodiment, in step (4), 10% sulfuric acid ethanol solution is sprayed and dried at 105°C until the spots are clear. In a specific embodiment, step (1) is to take 0.5g of the finished granules, dissolve them in 5ml of water, extract them twice with n-butanol, each time with 10ml, combine the extracts, evaporated in a water bath, and dissolve the residue in 2ml of methanol as the test solution; step (2) is to take another 0.2g of the positive control medicinal material, add an appropriate amount of water, reflux extract for 60 minutes, cool, filter, and concentrate the filtrate to 10ml, and take 5ml of the negative sample solution without Forsythia suspensa, and extract them twice with n-butanol, each time with 15ml, combine the extracts, evaporated in a water bath, and dissolve the residue in 2ml of methanol as the control medicinal material solution.

[0067] In one embodiment, for Coptis chinensis, steps (1) and (2) are added with n-butanol for extraction twice, the extracts are combined, evaporated in a water bath, and the residue is dissolved in methanol as the test solution. In another embodiment, in step (4), the granules are placed in a development cylinder pre-saturated with concentrated ammonia solution for 20 minutes, developed, taken out, dried, and inspected under a 365nm ultraviolet lamp; in a specific embodiment, step (1) is to take 0.5g of the finished granules, dissolve them in 5ml of water, extract them twice with n-butanol, 10ml each time, combine the extracts, evaporated in a water bath, and the residue is dissolved in 2ml of methanol as the test solution; step (2) is to take 0.25g each of the positive and negative control medicinal materials, add n-butanol, ultrasonically treat for 30min, filter, evaporate the filtrate to dryness, and dissolve the residue in 2ml of methanol as the control medicinal material solution.

[0068] In one embodiment, for licorice, steps (1) and (2) are added with n-butanol for extraction twice, the extracts are combined, evaporated to dryness in a water bath, and the residue is dissolved in methanol as the test solution. In another embodiment, in step (4), 10% sulfuric acid ethanol solution is sprayed, heated at 105°C until the spots are clear, and inspected under a 365nm ultraviolet lamp; in a specific embodiment, 0.1g of the positive licorice in step (2) is taken, an appropriate amount of water is added, refluxed for 60 minutes, cooled, filtered, and the filtrate is concentrated to 20ml, extracted twice with n-butanol, 15ml each time, the extracts are combined, evaporated to dryness in a water bath, and the residue is dissolved in 2ml of methanol as the control medicinal material solution.

[0069] In any of the above embodiments, the traditional Chinese medicine composition comprises the following components in parts by weight: 19-21 parts of Forsythia suspensa, 14-16 parts of Pinellia ternata, 11-13 parts of Radix Glycyrrhizae, 11-13 parts of Radix Bupleuri, 9-11 parts of Codonopsis pilosula, 7-9 parts of dried ginger, 5-7 parts of Scutellaria baicalensis, 4-6 parts of Coptis chinensis, 5-7 parts of Rhubarb, and 18-22 parts of jujube.

[0070] In another embodiment, the Chinese medicine composition includes the following components in parts by weight: 20 parts of Forsythia suspensa, 15 parts of Pinellia ternata, 12 parts of Radix Glycyrrhizae, 12 parts of Radix Bupleuri, 10 parts of Codonopsis pilosula, 8 parts of Dried Ginger, 6 parts of Scutellaria baicalensis, 5 parts of Coptis chinensis, 6 parts of Rhubarb, and 20 parts of Jujube.

[0071] In other embodiments, the traditional Chinese medicine composition further comprises the components in the above-mentioned weight ratios, and pharmaceutically acceptable excipients.

[0072] In a preferred embodiment, the traditional Chinese medicine composition is an oral preparation, preferably granules, tablets, capsules, pills, powders, etc.

[0073] In another preferred embodiment, the excipients of the formulation are disintegrants, fillers, binders, lubricants and / or bases.

[0074] Furthermore, the disintegrant includes one or more of starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinyl pyrrolidone, low-substituted hydroxypropyl cellulose, and cross-linked sodium carboxymethyl cellulose;

[0075] Furthermore, the filler includes one or more of starch, mannitol, chitin, pregelatinized starch, lactose, microcrystalline cellulose, and sucrose;

[0076] Furthermore, the lubricant includes one or more of sodium lauryl sulfate, magnesium stearate, talc, and silicon dioxide;

[0077] Furthermore, the adhesive comprises one or more of starch slurry, hydroxypropyl methylcellulose and polyvinyl pyrrolidone;

[0078] Furthermore, the matrix includes one of maltodextrin or PEG4000 (polyethylene glycol 4000) and PEG6000 (polyethylene glycol 6000);

[0079] In any of the above embodiments, the silica gel G thin layer plate is selected from German Merck silica gel G plate or Qingdao Ocean Chemical silica gel G plate. In a preferred embodiment, the silica gel G thin layer plate is selected from German Merck silica gel G plate.

[0080] The third principle of the present invention is to further adjust and optimize process parameters such as reagent extraction parameters after determining the identification developer group, and verify the stability and repeatability of the optimized developer under different quality control environments (such as high and low temperature, high humidity and low humidity, etc.).

[0081] A fourth object of the present invention is to provide a method for preparing the above-mentioned traditional Chinese medicine composition, comprising decocting the above-mentioned 10 herbs twice with water, filtering, combining the decoctions, concentrating the filtrate under reduced pressure to a clear paste, adding an appropriate amount of maltodextrin, drying or crushing, and preparing granules of the traditional Chinese medicine composition, performing quality control on the prepared granules, and guiding standardized production of the product based on the quality control results, wherein the method is characterized in that:

[0082] (1) taking the dried paste powder, dissolving it in water, adding an extractant to extract it, and combining the extracts;

[0083] (2) Evaporate the extract to dryness in a water bath, and dissolve the residue in methanol as the test solution; prepare positive and negative control solutions in parallel using the same method;

[0084] (3) Pipette the test solution, positive control solution, and negative control solution separately, place them on the same silica gel G thin layer plate, develop them on the thin layer identification developer, and take them out;

[0085] (4) Add color developing solution, heat with hot air until the spots are clearly colored, and inspect under ultraviolet light;

[0086] (5) Compare the spots of the test sample, positive control, and negative control. If the test sample and the positive control have spots of the same color at the corresponding positions, and there are no interfering spots on the negative control, then the test sample is judged to contain the drug component of the same or similar quality as the positive control;

[0087] For Forsythia suspensa, chloroform and methanol were mixed in a volume ratio of 8:1 as the developing solvent;

[0088] For Coptis chinensis, mix toluene: ethyl acetate: methanol: isopropanol: water in a volume ratio of 6:3:2:1.5:0.3 as a developing agent and place it in a developing cylinder that has been pre-saturated with concentrated ammonia solution for 20 minutes;

[0089] For licorice, ethyl acetate: glacial acetic acid: water was mixed in a volume ratio of 15:2:2 as the developing solvent;

[0090] The traditional Chinese medicine composition comprises the following components in parts by weight: 20 parts of forsythia, 15 parts of pinellia, 12 parts of roasted liquorice, 12 parts of bupleurum, 10 parts of codonopsis, 8 parts of dried ginger, 6 parts of scutellaria, 5 parts of yacon, 6 parts of cooked rhubarb, and 20 parts of jujube, and can be used to treat oral ulcers, acute and chronic gastroenteritis, and fox-induced madness (Behcet's syndrome).

[0091] In one embodiment, for Forsythia suspensa, steps (1) and (2) are extracted twice with n-butanol, the extracts are combined, evaporated in a water bath, and the residue is dissolved in methanol as the test solution. Step (2) is to take another 0.2g of positive control medicinal material, add an appropriate amount of water, reflux extract for 60 minutes, cool, filter, and concentrate the filtrate to 10ml, and take 5ml of negative sample solution without Forsythia suspensa, and extract the two with n-butanol twice, each time 15ml, combine the extracts, evaporated in a water bath, and dissolve the residue in 2ml of methanol as the control medicinal material solution. In another embodiment, step (3) is to take 6μl of the test solution and 4μl of the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, and use chloroform: methanol mixed in a volume ratio of 8:1 as the developing solvent for development; in another embodiment, step (4) is sprayed with 10% sulfuric acid ethanol solution and dried at 105℃ until the spots are clear. In a specific embodiment, step (1) is to take 0.5 g of the finished granules, dissolve them in 5 ml of water, extract them twice with 10 ml of n-butanol each time, combine the extracts, evaporate to dryness in a water bath, and dissolve the residue in 2 ml of methanol as the test solution.

[0092] In one embodiment, for Coptis chinensis, steps (1) and (2) are performed by adding n-butanol for extraction twice, the extracts are combined, evaporated to dryness in a water bath, and the residue is dissolved in methanol as the test solution. In another embodiment, in step (3), 3 μl of the test solution and 2 μl of the Coptis chinensis control medicinal material solution are respectively applied to the same silica gel G thin layer plate, and a mixture of toluene: ethyl acetate: methanol: isopropanol: water is adjusted to a volume ratio of 6:3:2:1.5:0.3 as a developing agent, and the plate is placed in a developing cylinder pre-saturated with concentrated ammonia test solution for 20 minutes for development; in another embodiment, in step (4), the plate is placed in a developing cylinder pre-saturated with concentrated ammonia test solution for 20 minutes, developed, taken out, dried, and placed under a 365nm ultraviolet lamp for inspection. In a specific embodiment, the step (1) is to take 0.5 g of the finished granules, add 5 ml of water to dissolve, add n-butanol to extract twice, 10 ml each time, combine the extracts, evaporate to dryness in a water bath, and dissolve the residue in 2 ml of methanol as the test solution; the step (2) is to take 0.25 g each of the positive and negative control medicinal materials, add n-butanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate to dryness, and dissolve the residue in 2 ml of methanol as the control medicinal material solution.

[0093] In one embodiment, for licorice, steps (1) and (2) are performed by adding n-butanol for extraction twice, the extracts are combined, evaporated to dryness in a water bath, and the residue is dissolved in methanol as the test solution. In another embodiment, in step (3), 7 μl of the test solution and 3 μl of the control medicinal material solution are respectively aspirated and spotted on the same silica gel G thin layer plate, and developed using a mixture of ethyl acetate: glacial acetic acid: water in a volume ratio of 15:2:2 as a developing solvent.

[0094] In another embodiment, in step (4), the sample is sprayed with 10% sulfuric acid ethanol solution, heated at 105°C until the spots are clear, and then inspected under a 365nm ultraviolet lamp. In a specific embodiment, 0.1g of the positive licorice sample in step (2) is added with an appropriate amount of water and refluxed for 60 minutes. The sample is cooled, filtered, and the filtrate is concentrated to 20ml. The sample is extracted twice with n-butanol, 15ml each time. The extracts are combined and evaporated to dryness in a water bath. The residue is dissolved in 2ml of methanol as a control medicinal material solution.

[0095] In any of the above embodiments, the preparation step of the traditional Chinese medicine composition comprises weighing the following traditional Chinese medicine components in parts by weight: 19-21 parts of old forsythia, 14-16 parts of ginger pinellia, 11-13 parts of roasted licorice, 11-13 parts of bupleurum, 9-11 parts of codonopsis, 7-9 parts of dried ginger, 5-7 parts of kushen, 4-6 parts of yalian, 5-7 parts of cooked rhubarb, and 18-22 parts of jujube;

[0096] (1) Decoction the above raw materials with water twice, each time for 0.5-1.0 hours, filter, and combine the decoctions;

[0097] (2) Concentrating the filtrate under reduced pressure at 60°C to a clear paste with a relative density of 1.03 to 1.10;

[0098] (3) adding an appropriate amount of matrix to the clear paste obtained in step (3), stirring evenly, and drying and granulating to obtain a clear paste powder;

[0099] (4) Monitor the quality of the dried paste powder and guide standardized production of the product based on the quality monitoring results;

[0100] (5) Based on the quality control results, different disintegrants, fillers, binders, lubricants and / or bases are added to the dry extract powder that meets the quality standards to prepare granules, tablets, capsules, pills or powders;

[0101] Furthermore, in step (2), 8-10 times the amount of water is added for the first time, soaked for 0.5 hours, and boiled for 1.0 hours; 6-8 times the amount of water is added for the second time, boiled for 0.5 hours, the decoctions are combined, and filtered;

[0102] Furthermore, in step (3), the filtrate obtained in step (2) is concentrated under reduced pressure at a temperature of 60°C ± 5°C to obtain a clear paste with a density of 1.03 to 1.05, and the clear paste is centrifuged at 10,000 rpm / min;

[0103] Furthermore, in step (4), the above-mentioned centrifuged clear paste is taken, an appropriate amount of maltodextrin is added, dissolved, mixed, and spray-dried to obtain a dry extract powder;

[0104] Furthermore, the quality monitoring of step (5) includes randomly extracting granulated samples, performing thin layer chromatography identification using the aforementioned developer group, testing and evaluating the quality of the product, and judging whether the product quality is qualified, thereby determining whether to continue or interrupt the production process of the product.

[0105] In any of the above embodiments, the silica gel G thin layer plate is selected from German Merck silica gel G plate or Qingdao Ocean Chemical silica gel G plate. In a preferred embodiment, the silica gel G thin layer plate is selected from German Merck silica gel G plate.

[0106] The fourth principle of the present invention is to introduce the above-determined quality control results into the production of the Chinese medicine composition, thereby providing a standardized production method that can meet product quality requirements.

[0107] The fifth object of the present invention is to provide a Chinese medicine composition prepared by any of the above preparation methods.

[0108] Compared with the traditional process, the present invention has the following advantages:

[0109] 1. The composition of the present invention is prepared by extracting only ten medicinal materials through traditional technology. It has the effects of invigorating qi and harmonizing the stomach, eliminating abdominal distension and stopping vomiting, clearing away upper heat and warming the lower body, and is used to treat oral erosion (oral ulcer), especially for the treatment of recurrent oral ulcer.

[0110] 2. The present invention can treat stomach bloating, rumbling in the abdomen, dry retching, irritability, diarrhea, indigestion, etc., and it also has certain effects on acute and chronic gastroenteritis and fox-induced madness (Behcet's syndrome).

[0111] 3. The present invention also refers to the standards of the main medicinal materials in the prescriptions included in the "Chinese Pharmacopoeia", the relevant standards of related Chinese medicine preparations and literature, and conducts bold exploration and attempts, abandoning the quality control of some ingredients that are not necessary or have little impact on product quality, selecting a standard method different from the standard method recommended by the "Chinese Pharmacopoeia", and conducting combination tests on various developing agents for the main components of Forsythia suspensa, Licorice and Coptis chinensis, so as to achieve a small number of identification developing agent groups to simultaneously detect multiple Chinese medicine ingredients, while ensuring the identification effect and optimizing and simplifying the quality control process.

[0112] 4. As for this method, the invention boldly explored and tried various traditional Chinese medicine ingredients of the present invention, discarded the quality control of some ingredients that are not necessary or have little impact on product quality, and conducted combination tests on various developing agents for the main ingredients such as Forsythia suspensa, Licorice, and Coptis chinensis, so as to achieve a small number of identification developing agent groups to simultaneously detect multiple traditional Chinese medicine ingredients, while ensuring the identification effect and optimizing and simplifying the quality control process;

[0113] 5. After the identification developer group is determined, further adjust and optimize the process parameters such as the reagent extraction parameters, and verify the stability and repeatability of the optimized developer under different quality control environments (such as high and low temperature, high humidity and low humidity, etc.);

[0114] 6. Introduce the above-determined quality control results into the production of the Chinese medicine composition, thereby providing a standardized production method that can meet product quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0115] Figure 1 : Thin layer chromatography spectrum of the forsythia identification of the traditional Chinese medicine composition granules of the present invention in Example 2, wherein 1 is forsythia control medicinal material solution I; 2 is forsythia negative control solution I; 3 is test sample (20190901) solution I; 4 is forsythia negative control solution II; 5 is forsythia control medicinal material solution II; 6 is test sample (20190901) solution II;

[0116] Figure 2 : The thin layer chromatography spectrum of the sample amount of the forsythia suspensa identification of the traditional Chinese medicine composition granules of the present invention in Example 2, wherein 1-3 are 2μl, 4μl, and 6μl of the forsythia suspensa control medicinal material solution II; 4-6 are 2μl, 4μl, and 6μl of the test sample (20190901) solution II;

[0117] Figure 3 : The identification specificity of the Chinese medicine composition granules of the present invention forsythia suspensa in Example 2 was investigated by thin layer chromatography, wherein 1 is a forsythia suspensa control medicinal material solution, 2 is a forsythia suspensa negative control solution, 3 is 20190901, 4 is 20190902, and 5 is 20190903;

[0118] Figure 4 : The thin layer chromatography patterns of the TCM composition granules of the present invention forsythiae at different temperatures and humidities were examined in Example 2. The composition of the present invention (sample volume: 4 μl for control, 6 μl for negative control and test sample) was as follows: 1 was a control forsythiae; 2 was a negative forsythiae; 3 was 20190901; 4 was 20190902; and 5 was 20190903. In addition, A: 27°C / RH94%; B: 26°C / RH28%; C: 4°C / RH84%;

[0119] Figure 5 : The thin layer chromatography patterns of different thin layer plates for identifying the Chinese medicine composition granules of Forsythia suspensa of the present invention in Example 2 were examined. The composition of the present invention (spot volume: 4 μl for control, 6 μl for negative control and test sample), wherein 1 is a Forsythia suspensa control medicinal material; 2 is a Forsythia suspensa negative; 3 is 20190901; 4 is 20190902; 5 is 20190903; in addition, A: Qingdao plate (26°C / RH51%), B: Merck plate (26°C / RH51%);

[0120] Figure 6 : Thin layer chromatography of three batches of samples of the Chinese medicine composition granules of the present invention, Forsythia suspensa test in Example 2, wherein 1 is a Forsythia suspensa control medicinal material solution, 2 is a Forsythia suspensa negative control solution, 3 is a test sample (20190901); 4 is a test sample (20190902); 5 is a test sample (20190903); In addition, T: 26 ° C, RH: 51%, sample volume: 4 μl for control, 6 μl for negative control and test sample, Qingdao Marine Silica G prefabricated plate;

[0121] Figure 7 : Thin layer chromatography spectrum of identification of the Chinese medicine composition granules of Coptis chinensis of the present invention in Example 3, wherein 1 is Coptis chinensis control medicinal material solution I, 2 is Coptis chinensis negative control solution I, 3 is test sample (20190901) solution I, 4 is Coptis chinensis control medicinal material solution II, 5 is Coptis chinensis negative control solution II, and 6 is test sample (20190901) solution II; in addition, T: 26 ° C, RH: 53%, sample volume: 2 μl, Qingdao Marine Silica G prefabricated plate;

[0122] Figure 8 : In Example 3, the identification spot sample amount of the Chinese medicine composition granules of the present invention, Coptis chinensis, was examined by thin layer chromatography, wherein 1-3 were Coptis chinensis control medicinal material solution II 1 μl, 2 μl, and 3 μl, and 4-6 were test sample (20190901) solution II 2 μl, 3 μl, and 4 μl; in addition, T: 26°C, RH: 53%, Qingdao Marine Silica G prefabricated plate.

[0123] Figure 9 : Thin layer chromatography spectrum of the identification specificity of the Chinese medicine composition granules of Coptis chinensis of the present invention, wherein 1 is the Coptis chinensis control medicinal material solution, 2 is the Coptis chinensis negative control solution, 3 is the test sample (20190901); 4 is the test sample (20190902); 5 is the test sample (20190903); In addition, T: 26 ° C, RH: 53%, sample volume: control 2 μl, negative control and test sample 3 μl, Qingdao Ocean Silica G prefabricated plate;

[0124] Figure 10 : Thin layer chromatography spectra of the identification of the Chinese medicine composition granules of Coptis chinensis of the present invention at different temperatures and humidities in Example 3 (sample volume: control 4 μl, test sample 6 μl), wherein 1 is the control medicinal material of Coptis chinensis; 2 is the test sample (20190901); 3 is the test sample (20190902); 4 is the test sample (20190903); In addition, A: 27°C / RH94%, B: 26°C / RH28%, C: 4°C / RH84%;

[0125] Figure 11: In Example 3, the thin layer chromatography spectra of the Chinese medicine composition granules of the present invention were examined on different thin layer plates, wherein 1 is the control medicinal material of Coptis chinensis; 2 is the test sample (20190901); 3 is the test sample (20190902); 4 is the test sample (20190903); In addition, T: 26 ° C, RH: 53%, sample volume: control 2 μl, test sample 3 μl, Merck silica gel G precast plate;

[0126] Figure 12 : Thin layer chromatography spectra of three batches of samples of the Chinese medicine composition granules of the present invention, Coptis chinensis test in Example 3, wherein 1 is a Coptis chinensis control medicinal material solution, 2 is a Coptis chinensis negative control solution, 3 is a test sample (20190901), 4 is a test sample (20190902), and 5 is a test sample (20190903); In addition, T: 26 ° C, RH: 53%, sample volume: 2 μl for control, 3 μl for negative control and test sample, Qingdao Marine Silica G prefabricated plate;

[0127] Figure 13 : Thin-layer chromatography of the licorice root identification of the traditional Chinese medicine composition granules of the present invention in Example 4, wherein 1 is licorice control medicinal material solution I, 2 is licorice negative control solution I, 3 is test sample (20190901) solution I, 4 is licorice control medicinal material solution II, 5 is licorice negative control solution II, and 6 is test sample (20190901) solution II; In addition, temperature: 26°C, relative humidity: 53%, sample volume: 3μl, Qingdao Ocean Silica G precast plate. A: plate without 1% NaOH treatment; B: plate treated with 1% NaOH;

[0128] Figure 14 : The ethyl acetate-glacial acetic acid-water (15:2:2) expansion system in Example 4, wherein 1 is licorice control medicinal material solution I, 2 is licorice negative control solution I, 3 is test sample (20190901) solution I, 4 is licorice control medicinal material solution II, 5 is licorice negative control solution II, and 6 is test sample (20190901) solution II; in addition, T: 26°C, RH: 53%, sample volume: 3 μl, Qingdao Ocean Silica G prefabricated plate;

[0129] Figure 15 : In Example 4, the identification of the Chinese medicine composition granules of the present invention was examined by spotting the sample in a thin layer chromatographic pattern, wherein 1-3 were 1 μl, 3 μl, and 5 μl of licorice control medicinal material solution II, and 4-6 were 3 μl, 5 μl, and 7 μl of the test sample (20190901) solution II; in addition, T: 26°C, RH: 53%, spotting volume: 3 μl, and Qingdao Ocean Silica G prefabricated plate;

[0130] Figure 16: Thin layer chromatography spectrum of the identification specificity of the licorice granules of the Chinese medicine composition of the present invention in Example 4, wherein 1 is a licorice control medicinal material solution, 2 is a licorice negative control solution, 3 is a test sample (20190901), 4 is a test sample (20190902), and 5 is a test sample (20190903); in addition, T: 26 ° C, RH: 53%, spotting volume: 3 μl for control, 7 μl for negative control and test sample, Qingdao Marine Silica G prefabricated plate;

[0131] Figure 17 : Thin layer chromatography of the granular licorice composition of the present invention at different temperatures and humidities in Example 4 (sample volume: control 3 μl, test sample 7 μl), wherein 1 is a licorice control medicinal material, 2 is a test sample (20190901), 3 is a test sample (20190902), and 4 is a test sample (20190903); in addition, A: 28°C / RH94%, B: 26°C / RH28%, and C: 4°C / RH84%;

[0132] Figure 18 : The thin layer chromatography spectra of different thin layer plates for identifying the granular licorice of the Chinese medicine composition of the present invention in Example 4 were examined, wherein 1 is a licorice control medicinal material, 2 is a test sample (20190901), 3 is a test sample (20190902), and 4 is a test sample (20190903); in addition, T: 26 ° C, RH: 53%, sample volume: control 3 μl, test sample 7 μl, Merck silica gel G precast plate;

[0133] Figure 19 : Thin layer chromatography of three batches of samples of the Chinese medicine composition granules of the present invention, liquorice test in Example 4, wherein 1 is a liquorice control medicinal material solution, 2 is a liquorice negative control solution, 3 is a test sample (20190901), 4 is a test sample (20190902, and 5 is a test sample (20190903); in addition, T: 26°C, RH: 53%, sample volume: 3 μl for control, 7 μl for negative control and test sample, Qingdao Marine Silica G prefabricated plate;

[0134] Figure 20 : Example 6 relates to a schematic flow chart of the preparation of Qingfei Paidu Decoction for quality inspection. DETAILED DESCRIPTION

[0135] The present invention will be further described in detail with reference to the following specific examples and accompanying drawings. The protection content of the present invention is not limited to the following examples. Without departing from the spirit and scope of the inventive concept, any changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the appended claims. The process, conditions, reagents, experimental methods, etc. for implementing the present invention, except for the contents specifically mentioned below, are all common knowledge and common common sense in the art and are not particularly limited in the present invention.

[0136] Example 1: Test equipment and raw materials

[0137] 1. Instruments and test drugs

[0138] 1.1 Instrument

[0139] Thin layer automatic imaging instrument (CAMAG TLCⅤISUALIZER2, CAMAG, Switzerland), thin layer plate heating plate (CAMAG TLC PLATE HEATERⅢ, CAMAG, Switzerland), electronic balance (METTLER ME2002E, Mettler-Toledo Instrument (Shanghai) Co., Ltd.), electronic balance (METTLER MS 204TS, Mettler-Toledo Instrument (Shanghai) Co., Ltd.), ultrasonic cleaner (KQ-300DE, Kunshan Ultrasonic Instrument Co., Ltd.), medical centrifuge (Xiangyi H1850, Hunan Xiangyi Laboratory Instrument Development Co., Ltd.), constant temperature water bath (HWS-26), planar chromatography spotter (SPDY-1A, Nanjing Maikeliite Scientific Instrument Co., Ltd.), silica gel G thin layer precast plate (Qingdao Ocean Chemical Plant), silica gel G thin layer precast plate (Merck, Germany).

[0140] 1.2 Test drugs and reagents

[0141] Forsythia suspensa (batch number: 120955-201309), Glycyrrhiza uralensis (batch number: 120904-201519), and Coptis chinensis (batch number: 120913-201611) were all purchased from the China Food and Drug Administration.

[0142] The granules of the Chinese medicine composition of the present invention (batch numbers: 20190901, 20190902, 20190903) are provided by Guangdong Zhidao Pharmaceutical Technology Co., Ltd.

[0143] Negative control: According to the prescription: Forsythia suspensa 2g, Pinellia ternata 1.5g, Licorice 1.2g, Scutellaria baicalensis 0.6g, Coptis chinensis 0.5g, Dry ginger 0.8g, Jujube 2g, Codonopsis pilosula 1g, Rhubarb 0.6g, Bupleurum 1.2g, weigh the prescription amounts without Forsythia suspensa, Licorice Root and Coptis chinensis respectively, decoct them according to the preparation method specified in the standard, and concentrate to 60ml to obtain the negative controls without Forsythia suspensa, Licorice Root and Coptis chinensis.

[0144] Example 2: Thin layer chromatography analysis to determine the components of Forsythia suspensa

[0145] 1. Preparation of Forsythia suspensa Solution

[0146] 1.1 Preparation of test solution

[0147] (1) Take 0.5 g of this product (batch number: 20190901), add 5 ml of hot water to dissolve it, extract it twice with 10 ml of ethyl acetate each time, combine the ethyl acetate extracts, evaporate to dryness, and dissolve the residue with 2 ml of methanol to obtain test solution I.

[0148] (2) Take 0.5 g of this product (batch number: 20190901), add 5 ml of hot water to dissolve it, extract it twice with 10 ml of n-butanol each time, combine the n-butanol extracts, evaporate to dryness, and dissolve the residue with 2 ml of methanol to obtain test solution II.

[0149] 1.2 Preparation of control medicinal material solution

[0150] (1) Take 0.2 g of Forsythia suspensa reference medicinal material powder, add appropriate amount of water, reflux extraction for 1 hour, filter, concentrate the filtrate to 10 ml, extract with ethyl acetate twice, 15 ml each time, combine the extracts, evaporate to dryness, and dissolve the residue in 2 ml of methanol to obtain Forsythia suspensa reference medicinal material solution I.

[0151] (2) Take 0.2 g of Forsythia suspensa reference medicinal material powder, add appropriate amount of water, reflux extraction for 1 hour, filter, concentrate the filtrate to 10 ml, extract with n-butanol twice, 15 ml each time, combine the extracts, evaporate to dryness, and dissolve the residue in 2 ml of methanol to obtain Forsythia suspensa reference medicinal material solution II.

[0152] 1.3 Preparation of negative control solution

[0153] (1) Take 5 ml of the negative sample solution lacking Forsythia suspensa and extract it twice with 15 ml of ethyl acetate each time. Combine the ethyl acetate extracts and evaporate to dryness in a water bath. Dissolve the residue in 2 ml of methanol to obtain Forsythia suspensa negative control solution I.

[0154] (2) Take 5 ml of the negative sample solution without Forsythia suspensa, extract it twice with 15 ml of n-butanol each time, combine the n-butanol extracts, evaporate to dryness in a water bath, and dissolve the residue in 2 ml of methanol to obtain Forsythia suspensa negative control solution II.

[0155] 2. Thin layer board

[0156] It is planned to use the silica gel G prefabricated panels from Shandong Qingdao Ocean Chemical Plant to carry out experimental research.

[0157] 3. Results

[0158] Develop according to the above development conditions, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ until the spots are clear, where T: 26℃, RH: 51%, Qingdao Ocean Silica G prefabricated plate, the same below.

[0159] See the results Figure 1 .

[0160] The results show:

[0161] ①This expansion system can identify the medicinal materials of Forsythia suspensa in the sample;

[0162] ② Extraction with n-butanol is more complete than extraction with ethyl acetate.

[0163] Conclusion: This development system has strong specificity and the negative control has no interference, so it is proposed to be included in the standard.

[0164] 4. Sample quantity inspection

[0165] 2μl, 4μl, and 6μl of the test solution II and 2μl, 4μl, and 6μl of the Forsythia suspensa control solution II were respectively taken and spotted on the same silica gel G thin layer prefabricated plate. The plate was developed according to the above development method, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105℃ until the spots were clear. The results are shown in the table. Figure 2 .

[0166] The results showed that it was more appropriate to spot 6μl of the test solution II and 4μl of the Forsythia suspensa control medicinal material solution II.

[0167] 5. The present invention confirms the method of thin layer identification of Forsythia suspensa

[0168] Dissolve 0.5g of this product in 5ml of hot water. Extract twice with n-butanol (10ml each time). Combine the n-butanol extracts, evaporate to dryness, and dissolve the residue in 2ml of methanol to prepare the test solution. Separately, extract 0.2g of Forsythia suspensa control material with an appropriate amount of water. Reflux and extract for 1 hour. Filter. Concentrate the filtrate to 10ml and extract twice with n-butanol (15ml each time). Combine the n-butanol extracts, evaporate to dryness, and dissolve in 2ml of methanol to prepare the control solution. For thin-layer chromatography, spot 6μl of the test solution and 4μl of the control solution on the same silica gel G thin-layer plate using a mixture of chloroform and methanol (8:1 by volume) as the developing solvent. Develop, remove, air-dry, spray with 10% ethanolic sulfuric acid, and heat at 105°C until the spots are clear. The test sample chromatogram should show spots of the same color at the corresponding positions on the Forsythia suspensa control chromatogram.

[0169] 6. Verification of the Thin Layer Identification Method of Forsythia suspensa of the Present Invention

[0170] (1) Specificity inspection

[0171] Take 6μl of three batches of test sample solution, 4μl of Forsythia suspensa control medicinal material solution, and 6μl of Forsythia suspensa negative control solution, and perform identification under the same chromatographic conditions as above. The results are shown in Figure 3 ;

[0172] The results showed that there was no interference with the negative test and the method was feasible.

[0173] (2) Comparison of different temperatures and humidity

[0174] According to the proposed thin layer identification method, the samples were developed at different temperatures and humidities. The results can be found in Figure 4 .

[0175] The results showed that under different temperature and humidity environments, the chromatographic behavior of the test samples was consistent with that of the main spots of the control medicinal materials.

[0176] (3) Comparison of different thin layer plates

[0177] According to the proposed thin layer identification method, prefabricated panels from different manufacturers were used for development. The results can be found in Figure 5 .

[0178] The results showed that when developed on different thin-layer plates, the test samples all showed chromatographic behaviors consistent with the main spots of the control medicinal materials, and the spots were clearly separated, and all of them could be used for testing this method.

[0179] (4) Sample inspection

[0180] Take three batches of samples and test them according to the above confirmed method. The results are shown in Figure 6 .

[0181] (5) Conclusion

[0182] The thin-layer method for identifying Forsythia suspensa by the present invention is confirmed to have no negative interference, and the chromatographic behavior is consistent with the main spot of the positive control medicinal material. The spots are clearly separated, and under different temperature and humidity environments, the chromatographic behavior of the test sample spectrum is consistent with the main spot of the control medicinal material, indicating that the method has good stability and high sensitivity and has good versatility for different identification environments.

[0183] Example 3: Thin layer chromatography analysis to determine the components of Coptis chinensis

[0184] 1. Preparation of Coptis chinensis Solution

[0185] 1.1 Preparation of test solution

[0186] (1) Take 0.5 g of this product (batch number: 20190901), add 5 ml of hot water to dissolve it, extract it twice with 10 ml of ethyl acetate each time, combine the ethyl acetate extracts, evaporate to dryness, and dissolve the residue with 2 ml of methanol to obtain test solution I.

[0187] (2) Take 0.5 g of this product (batch number: 20190901), add 5 ml of hot water to dissolve it, extract it twice with 10 ml of n-butanol each time, combine the n-butanol extracts, evaporate to dryness, and dissolve the residue with 2 ml of methanol to obtain test solution II.

[0188] 1.2 Preparation of control medicinal material solution

[0189] (1) Add 25 ml of ethyl acetate to 0.25 g of Rhizoma Coptidis reference medicinal material, ultrasonically treat for 30 min, filter, and use the filtrate as Rhizoma Coptidis reference medicinal material solution I;

[0190] (2) 0.25 g of Rhizoma Coptidis reference medicinal material was added with 25 ml of methanol, and the mixture was ultrasonically treated for 30 min. The mixture was filtered and the filtrate was used as Rhizoma Coptidis reference medicinal material solution II.

[0191] 1.3 Preparation of negative control solution

[0192] (1) Take 2.5 ml of the negative sample solution lacking Coptis chinensis, extract twice with 10 ml of ethyl acetate each time, combine the ethyl acetate extracts, evaporate to dryness in a water bath, and dissolve the residue in 2 ml of methanol to obtain Coptis chinensis negative control solution I;

[0193] (2) Take 2.5 ml of the negative sample solution lacking Coptis chinensis, extract twice with 10 ml of n-butanol each time, combine the n-butanol extracts, evaporate to dryness in a water bath, and dissolve the residue in 2 ml of methanol to obtain Coptis chinensis negative control solution II.

[0194] 2. Thin layer board

[0195] It is planned to use the silica gel G prefabricated panels from Shandong Qingdao Ocean Chemical Plant to carry out experimental research.

[0196] 3. Results

[0197] Expand according to the above expansion conditions, take out, dry, and examine under 365nm ultraviolet light. For the results, see Figure 7 .

[0198] The results show:

[0199] ①This expansion system can identify the medicinal material Coptis chinensis in the sample;

[0200] ② Ethyl acetate can hardly extract the components of Coptis chinensis, while n-butanol can extract them more fully.

[0201] Conclusion: This development system has strong specificity and the negative control has no interference, so it is proposed to be included in the standard.

[0202] 4. Sample quantity inspection

[0203] Pipette 1μl, 2μl, and 3μl of the test solution II under 3.2.1 and 1μl, 2μl, and 3μl of the Rhizoma Coptidis control solution II respectively, and spot them on the same silica gel G thin layer prefabricated plate. Develop it according to the above development method, remove it, dry it, and examine it under a 365nm ultraviolet lamp. For the results, see Figure 8 ;

[0204] The results showed that it was more appropriate to spot 3μl of the test solution II and 2μl of the Coptis chinensis control medicinal material solution II.

[0205] 5. The present invention confirms the method of thin layer identification of Coptis chinensis

[0206] Take 1 ml of the identification test sample solution II and dilute it to 2 ml with methanol to prepare the test sample solution. Separately, take 0.25 g of the Rhizoma Coptidis (Coptidis Rhizoma) control material and add 25 ml of methanol. Ultrasonicate for 30 minutes, filter, and use the filtrate as the Rhizoma Coptidis control material solution. Apply 3 μl of the test sample solution and 2 μl of the Rhizoma Coptidis control material solution to the same silica gel G thin-layer plate. Use a mixture of toluene, ethyl acetate, methanol, isopropanol, and water (6:3:2:1.5:0.3 by volume) as the developing solvent. Place the plate in a developing chamber presaturated with concentrated ammonia solution for 20 minutes. Develop, remove, air dry, and examine under a 365 nm UV lamp. The test sample chromatogram should produce a fluorescent spot of the same color as the Rhizoma Coptidis control chromatogram at the corresponding position.

[0207] 6. Method Validation

[0208] (1) Specificity inspection

[0209] Take 3μl of three batches of test sample solution, 2μl of Coptis chinensis control medicinal material solution, and 3μl of Coptis chinensis negative control solution, and perform identification under the same chromatographic conditions as above. The results are shown in Figure 9 ;

[0210] The results showed that there was no interference with the negative test and the method was feasible.

[0211] (2) Comparison of different temperatures and humidity

[0212] According to the proposed thin layer identification method, the samples were developed at different temperatures and humidities. The results can be found in Figure 10 ;

[0213] The results showed that under different temperature and humidity environments, the chromatographic behavior of the test samples was consistent with that of the main spots of the control medicinal materials.

[0214] (3) Comparison of different thin layer plates

[0215] According to the proposed thin layer identification method, the German Merck silica gel G thin layer prefabricated plate was used for development. The results can be found in Figure 11 ;

[0216] The results showed that when developed on different thin-layer plates, the test samples all showed chromatographic behaviors consistent with the main spots of the control medicinal materials, and the spots were clearly separated, and all of them could be used for testing this method.

[0217] (4) Sample inspection

[0218] Take three batches of samples and test them according to the above confirmed method. The results are shown in Figure 12 .

[0219] (5) Conclusion

[0220] The present invention adjusts toluene: ethyl acetate: methanol: isopropanol: water in a volume ratio of 6:3:2:1.5:0.3 to be mixed as a developing agent, thereby confirming a thin-layer identification method for Coptis chinensis, with no interference in the negative state, consistent chromatographic behavior with the main spot of the positive control medicinal material, clear spot separation, and consistent chromatographic behavior with the main spot of the control medicinal material in the test sample spectrum under different temperature and humidity environments, indicating that the method has good stability and high sensitivity and has good versatility for different identification environments.

[0221] Example 4: Thin-layer chromatography analysis of liquorice components

[0222] 1. Preparation of Licorice Solution

[0223] 1.1 Preparation of test solution

[0224] (1) Take 0.5 g of this product (batch number: 20190901), add 5 ml of hot water to dissolve it, extract it twice with 10 ml of ethyl acetate each time, combine the ethyl acetate extracts, evaporate to dryness, and dissolve the residue with 2 ml of methanol to obtain test solution I;

[0225] (2) Take 0.5 g of this product (batch number: 20190901), add 5 ml of hot water to dissolve it, extract it twice with 10 ml of n-butanol each time, combine the n-butanol extracts, evaporate to dryness, and dissolve the residue with 2 ml of methanol to obtain test solution II.

[0226] 1.2 Preparation of control medicinal material solution

[0227] (1) Take 0.1 g of licorice control medicinal material powder, add appropriate amount of water, reflux extraction for 1 hour, filter, concentrate the filtrate to 10 ml, extract with ethyl acetate twice, 15 ml each time, combine the extracts, evaporate to dryness, and dissolve the residue in 2 ml of methanol to obtain Forsythia suspensa control medicinal material solution I;

[0228] (2) Take 0.1 g of licorice control medicinal material powder, add appropriate amount of water, reflux extraction for 1 hour, filter, concentrate the filtrate to 10 ml, extract with n-butanol twice, 15 ml each time, combine the extracts, evaporate to dryness, and dissolve the residue in 2 ml of methanol to obtain licorice control medicinal material solution II.

[0229] 1.3 Preparation of negative control solution

[0230] (1) Take 5 ml of the negative sample solution lacking licorice, extract it twice with ethyl acetate, 15 ml each time, combine the ethyl acetate extracts, evaporate to dryness in a water bath, and dissolve the residue in 2 ml of methanol to obtain licorice negative control solution I;

[0231] (2) Take 5 ml of the negative sample solution lacking licorice, extract it twice with n-butanol, 15 ml each time, combine the n-butanol extracts, evaporate to dryness in a water bath, and dissolve the residue in 2 ml of methanol to obtain licorice negative control solution II.

[0232] 2. Thin layer board

[0233] It is planned to use the silica gel G prefabricated panels from Shandong Qingdao Ocean Chemical Plant to carry out experimental research.

[0234] 3. Results

[0235] (1) Mix ethyl acetate, glacial acetic acid, and water in a volume ratio of 15:2:2 as a developing agent. Develop, remove, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clear, and examine under a 365nm ultraviolet lamp. The results are shown in the table. Figure 13 ;

[0236] The results show:

[0237] ① This expansion system identified licorice, and the negative had interference;

[0238] ② n-Butanol extraction has more interfering components than ethyl acetate extraction, so ethyl acetate extraction should be used for sample processing;

[0239] ③ The thin layer prefabricated board is treated with 1% NaOH solution and not treated. The only difference is that the interference components are higher or lower, which is basically no difference in nature. Therefore, the prefabricated board does not need to be treated;

[0240] ④The expansion system needs further optimization.

[0241] Conclusion: The negative control of this development system has interference and needs further optimization.

[0242] (2) Optimize the above conditions and use ethyl acetate: glacial acetic acid: water in a volume ratio of 15:2:2 as the developing agent. Develop, remove, dry, and heat with 10% sulfuric acid ethanol solution at 105°C until the spots are clear. Place under 365nm ultraviolet light for inspection. The results are shown in the table. Figure 14 ;

[0243] The results show:

[0244] ①This expansion system can identify licorice, with negative and no interference;

[0245] ② n-Butanol extraction has more interfering components than ethyl acetate extraction, but the licorice spots are also clearer. Under this development system, the interfering components do not affect the identification of licorice. Therefore, n-butanol extraction should be used for sample processing, and test sample II under item 3.1.6 can be used directly.

[0246] 4. Sample quantity inspection

[0247] According to the thin layer chromatography method (Chinese Pharmacopoeia 2015 edition, Part IV, General Chapter 0502), 3 μl, 5 μl, and 7 μl of the test sample solution II under 3.3.1 and 1 μl, 3 μl, and 5 μl of the licorice control drug solution II were respectively aspirated and spotted on the same silica gel G thin layer precast plate. Develop according to the above developing method, remove, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C until the spots are clear, and examine under 365nm ultraviolet light. For the results, see [see ]. Figure 15 ;

[0248] The results showed that it was more appropriate to spot 7μl of the test solution II and 3μl of the licorice control medicinal material solution II.

[0249] 5. The present invention confirms the method of thin layer identification of liquorice

[0250] Take 0.1 g of the licorice control herb, add an appropriate amount of water, and reflux extract for 1 hour. Filter, concentrate the filtrate to 10 ml, and extract twice with n-butanol (15 ml each time). Combine the n-butanol extracts, evaporate to dryness, and dissolve in 2 ml of methanol to prepare the licorice control herb solution. Apply 7 μl of the test sample solution II and 3 μl of the control herb solution (described in 3.1.6) to the same silica gel G thin-layer plate. Develop with a mixture of ethyl acetate: glacial acetic acid: water (15:2:2 by volume). Develop, air dry, spray with 10% ethanolic sulfuric acid, heat at 105°C until the spots are clear, and examine under 365 nm UV light. The test sample chromatogram should show a spot of the same color or a fluorescent spot at the corresponding position in the chromatogram of the licorice control herb.

[0251] 6. Method Validation

[0252] (1) Specificity inspection

[0253] Take 7μl of three batches of test sample solution, 3μl of licorice control medicinal material solution, and 7μl of licorice negative control solution, and perform identification under the same chromatographic conditions as above. The results are shown in Figure 16 ;

[0254] The results showed that there was no interference with the negative test and the method was feasible.

[0255] (2) Comparison of different temperatures and humidity

[0256] According to the proposed thin layer identification method, the samples were developed at different temperatures and humidities. The results can be found in Figure 17 ;

[0257] The results showed that under different temperature and humidity environments, the chromatographic behavior of the test samples was consistent with that of the main spots of the control medicinal materials.

[0258] (3) Comparison of different thin layer plates

[0259] According to the proposed thin layer identification method, the German Merck silica gel G thin layer prefabricated plate was used for development. The results can be found in Figure 18 ;

[0260] The results showed that when developed on different thin-layer plates, the test samples all showed chromatographic behaviors consistent with the main spots of the control medicinal materials, and the spots were clearly separated, and all of them could be used for testing this method.

[0261] (4) Sample inspection

[0262] Take three batches of samples and test them according to the above confirmed method. The results are shown in Figure 19 ;

[0263] The results show that it meets the standard requirements.

[0264] (5) Conclusion

[0265] The method for thin-layer identification of licorice confirmed by the present invention has negative interference-free results, chromatographic behavior consistent with the main spot of the positive control medicinal material, clear spot separation, and chromatographic behavior consistent with the main spot of the control medicinal material in the test sample spectrum under different temperature and humidity environments, indicating that the method has good stability and high sensitivity and has good versatility for different identification environments.

[0266] Example 5: Research on small-scale extraction process

[0267] In this study, the corresponding physical object of the Chinese medicine composition granules of the present invention was used as a reference for the research on the small-scale extraction process, and the extraction process parameters closest to the corresponding physical object were taken as the optimal process parameters; the amount of water added and decoction time were used as the investigation factors, and a single-factor experiment was used to optimize the small-scale extraction process.

[0268] 1. Experimental Design

[0269] To simulate large-scale extraction conditions, an electric heating jacket was used for heating and reflux. The optimal extraction process parameters were determined by comparing two water addition ratios: 8 times the first decoction and 6 times the second decoction, and 6 times the first decoction and 6 times the second decoction, and three extraction time parameters of 60 minutes, 40 minutes, and 30 minutes. The experiment was designed as follows:

[0270] Table 1: Small-scale extraction process optimization experiment

[0271]

[0272] 2. Feeding amount

[0273] Forsythia suspensa 40g, raw licorice 24g, ginger and pinellia 30g, astragalus 12g, coptis 10g, dried ginger 16g, jujube 40g, cooked rhubarb 12g, codonopsis pilosula 20g, bupleurum 24g.

[0274] 3. Equipment and medicinal materials

[0275] Equipment: Heating mantle (98-1-B electronic temperature-controlled heating mantle); 5 L round-bottom flask; 1210 high-performance liquid chromatograph (Agilent Technologies, Inc.); ME204 electronic balance (Mettler-Toledo); HWS-26 electronic constant-temperature water bath (Shanghai Yiheng Scientific Instrument Co., Ltd.); H1850 high-speed centrifuge (Changsha Xiangrui Centrifuge Co., Ltd.);

[0276] Pieces:

[0277] Table 2: Information on raw materials for pilot test

[0278] Name of medicinal material Prescription quantity (g) batch number Origin Forsythia 40 2017062101 Yuncheng, Shanxi Province Ginger and Pinellia 30 160603 Sichuan Raw licorice 24 170801 Inner Mongolia Scutellaria baicalensis 12 19100709 Yuncheng, Shanxi Province Coptis chinensis 10 HL-YP-1910-15 Chongqing Shizhu dried ginger 16 GJ-YP-2005-41 Qujing City, Yunnan Province jujube 40 DZ-YP-2001-24 Ruoqiang County, Xinjiang Codonopsis pilosula 20 20200801 Longxi County, Gansu Province Cooked rhubarb 12 20180404 Gansu Bupleurum 24 CH-YP-1912-02 Gansu

[0279] 4. Sample Preparation

[0280] The experiment was carried out according to the experimental sequence set above. The heating reflux extraction was performed according to the water addition amount, extraction number, and extraction time set in each experiment. The two extracts were combined and the volume was recorded. The results are as follows:

[0281] Table 3: Small-scale extraction process optimization experiment

[0282]

[0283]

[0284] 5. Sample testing and analysis

[0285] According to the pharmacopoeia dry paste rate determination method and baicalin content determination method, each sample was measured and compared with the corresponding physical object prepared from the same batch of decoction pieces as a reference. The results are as follows:

[0286] Table 4: Results of small-scale extraction process research

[0287]

[0288] From the above results, it can be seen that among the test results of the experimental samples of each serial number, the experimental serial number 4 is closest to the content of the index component baicalin in the corresponding physical object, and the experimental serial number 2 is the second closest; the experimental serial number 4 is closest to the dry paste rate in the corresponding physical object, and the experimental serial number 2 is the second closest; considering that the extraction tank is heated at the periphery but not at the bottom during industrial production, the extraction effect at the bottom is affected, therefore, appropriately increasing the amount of water added is beneficial to compensate for the effect caused by the lack of heating at the bottom of the extraction tank, and after comprehensive consideration, the extraction conditions of experimental serial number 2 are selected as the pilot extraction process conditions for the Chinese medicine composition granules of the present invention, which are as follows:

[0289] Take the prescribed pieces, add water and extract twice. The first time, add 8 times the amount of water, soak for 30 minutes, heat and extract for 40 minutes, and filter; the second time, add 6 times the amount of water, extract for 30 minutes, filter, and combine the filtrate to obtain the product.

[0290] 6. Determination of pilot extraction process

[0291] Based on the above research results, the small-scale extraction process of the Chinese medicine composition granules of the present invention is determined as follows:

[0292] Take the prescribed pieces, add water and extract twice. The first time, add 8 times the amount of water, soak for 30 minutes, heat and extract for 40 minutes, and filter; the second time, add 6 times the amount of water, extract for 30 minutes, filter, and combine the filtrate to obtain the product.

[0293] 7. Choice of impurity removal method

[0294] Compare the two methods of impurity removal: high-speed centrifugal removal and screen filtration removal, and choose the appropriate impurity removal process.

[0295] 7.1 Experimental Comparison

[0296] Take the QXKYKL-T-02 sample under Item 2.3.4, centrifuge it at 10,000 rpm / min for 3 minutes in a desktop centrifuge, and compare it with the sample passed through a 200-mesh filter bag screen to examine the differences in solid content and baicalin content of the samples before and after impurity removal.

[0297] 7.2 Results and Analysis

[0298] Table 5: Comparison of different impurity removal methods

[0299]

[0300] From the above results, it can be seen that there is no difference in the baicalin content and solid content of the samples treated by the two impurity removal methods. Considering the convenience of large-scale production equipment, it is planned to use a 200-mesh sieve for filtration.

[0301] 8. Concentration temperature investigation

[0302] It is intended to compare the concentration of the granule extract of the Chinese medicine composition of the present invention at three temperature conditions: 60°C, 75°C, and 90°C, and to investigate the effects of different concentration temperatures on the extract.

[0303] 8.1 Experimental Comparison

[0304] Take three portions of QXKYKL-T-02 sample, each 500 ml, and concentrate the sample to 100 ml on a rotary evaporator at 60°C, 75°C, and 90°C, respectively. Then dilute the sample to 500 ml again and take samples for testing. The test indicator is baicalin content.

[0305] 8.2 Results and Analysis

[0306] Table 6: Effects of different concentration temperatures on samples

[0307]

[0308] From the above results, it can be seen that when the samples were concentrated at 60℃, 75℃, and 90℃, there was no significant difference in the baicalin content in the samples before and after concentration, indicating that the sample quality was improved when the sample was concentrated at 60℃ to 90℃. Therefore, combined with large-scale production practice, it is proposed to determine the concentration temperature to be below 80℃.

[0309] 9. Extraction process pilot test

[0310] Based on the extraction process parameters determined by the small-scale extraction process research, pilot-scale scale-up tests were conducted. Furthermore, considering that the small-scale extraction process used a heating reflux method, while the pilot-scale scale-up process used industrially produced extraction tanks, the following two sets of conditions were set up for comparison to determine the optimal production process.

[0311] Table 7: Comparison of pilot extraction process parameters

[0312]

[0313]

[0314] 9.1 Ingredients and Pieces

[0315] Table 8: Extraction process pilot study material information

[0316]

[0317]

[0318] 9.2 Extraction equipment

[0319] Table 9: Extraction equipment table

[0320]

[0321] 9.3 Extraction process

[0322] See also Figure 9 , process flow chart.

[0323] 9.4 Sample extraction process

[0324] Table 10: Extraction process table

[0325]

[0326]

[0327] 9.5 Extract test results

[0328] Table 11: Content of index components in pilot extracts

[0329]

[0330] From the above results, it can be seen that the pilot extraction samples under the two groups of pilot extraction conditions were compared with the corresponding physical materials prepared from the same batch of medicinal pieces. The baicalin content in the samples of group 1 was 3.2% higher than that of the physical material corresponding to the material standard. The baicalin content in the samples of group 2 was 3.2% higher than that of the physical material corresponding to the material standard. Therefore, the extraction conditions of group 1 were selected as the pilot extraction process parameters, which are as follows:

[0331] Take the prescribed decoction pieces and extract them with water twice. The first time, add 8 times the amount of water, soak for 30 minutes, heat and extract for 60 minutes, and filter; the second time, add 6 times the amount of water, heat and extract for 30 minutes, filter, and combine the filtrate.

[0332] 10. Concentration and drying

[0333] 10.1 Concentration

[0334] According to the results of the concentration temperature investigation under 2.5, the concentration of the extract at 60℃ to 90℃ maintains the stable content of the index components. In this pilot concentration study, a double-effect concentrator was used for concentration at a temperature of 75℃ to 80℃, which can maintain stable product quality.

[0335] 10.2 Drying

[0336] Common drying methods for traditional Chinese medicine preparations include vacuum oven drying, spray drying, microwave drying, and vacuum belt drying. In recent years, spray drying has become a common method for drying traditional Chinese medicine extracts because it shortens the sample's heating time and is particularly suitable for drying heat-sensitive substances.

[0337] Spray drying is a process that uses an atomizer to disperse a liquid into fine droplets, then rapidly evaporates the solvent in a hot drying medium to form a dry powder product. Spray drying can directly dry solutions, emulsions, and suspensions into powders, granules, hollow spheres, or pellets, eliminating the need for evaporation, separation, and pulverization. It offers high drying efficiency and minimal damage to active ingredients. The resulting extract powder is uniform, fine, low in water content, and highly soluble, making it suitable for large-scale industrial production and improving production efficiency and product quality.

[0338] In order to determine the drying method, spray drying and vacuum drying were compared to determine the appropriate drying method.

[0339] 10.2.1 Comparative Study of Spray Drying and Vacuum Drying

[0340] 28.8 kg of the concentrated extract from the pilot test in Group 1 was mixed with 2.6 kg of maltodextrin and stirred evenly. 100 ml of the sample was taken and dried in a vacuum oven at 60°C. Another portion of the sample was spray dried in a spray dryer and the dried samples were compared.

[0341] 10.2.2 Results and Analysis

[0342] Table 12: Comparison of different drying methods

[0343]

[0344] From the above results, it can be seen that the sample dried by spray drying has a slightly higher baicalin content than that by vacuum drying, and is in powder form, light in color, and has good water solubility; while the sample dried by vacuum drying has a slightly decreased baicalin content, but no significant difference is observed, the sample is dark in color, and takes a long time to dissolve. Combining the two drying methods, spray drying was selected as the drying method for this product.

[0345] 10.3 Selection of excipients

[0346] This study adopted a method of adding excipients to the concentrated extract and then drying it. Common excipients used in traditional Chinese medicine granules include soluble starch, dextrin, maltodextrin, lactose, xylitol, mannitol, and other excipients. Maltodextrin, also known as water-soluble dextrin or enzymatic dextrin, is a starch hydrolyzate made from starch through a low-level controlled enzymatic hydrolysis process, purification, and drying. It has low sweetness, good solubility, low moisture absorption, good stability, and difficulty in deterioration, and is widely used in pharmaceutical production. Preliminary experiments in this study showed that granules made from maltodextrin have advantages such as good solubility and clarity. Therefore, maltodextrin was selected as the excipient for this product.

[0347] 10.4 Granulation

[0348] The granulation method for this product is proposed to be boiling granulation, which is a relatively commonly used granulation method. It has the characteristics of adding relatively few excipients, good solubility of the prepared sample, light appearance color, and relatively simple operation.

[0349] 10.4.1 Equipment

[0350] Table 13: Granulation equipment

[0351]

[0352] 10.4.2 Parameters

[0353] Table 14: Granulation parameters

[0354] Granulation parameters Inlet air temperature 75℃ Material temperature range 45-60℃ Spray speed 15~30rpm(30~60ml / min) Atomization pressure 0.05pa

[0355] 10.4.3 Yield

[0356] The yield of this product is calculated according to the following formula, and the result is 91.2%

[0357]

[0358] 10.4.4 Sample determination

[0359] The samples were tested according to the finished product quality standards and met the requirements of the finished product quality standards. The main indicators are as follows:

[0360] Table: Main indicators of the granule samples of the Chinese medicine composition of the present invention

[0361] Table 15: Sample measurement results

[0362]

[0363] 11. Pilot process verification

[0364] Based on the experimental research process, three batches of pilot verification experiments were carried out:

[0365] For each batch, 22.8 kg of the prescribed slices were decocted twice. The first decoction was performed with 8 times the amount of water, soaked for 30 minutes, and decocted for 60 minutes. The second decoction was performed with 6 times the amount of water, and decocted for 30 minutes. The extract was filtered, and the filtrate was concentrated under reduced pressure to a specific gravity of 1.03-1.10 (60°C). An appropriate amount of maltodextrin was added, stirred, spray-dried, and granulated. The specific control conditions and parameters are as follows.

[0366] 11.1 Extraction process

[0367] The extraction process and experimental results are shown in the table below:

[0368] Table 16: Extraction process and results

[0369]

[0370] From the above results, it can be seen that the extraction process is smooth, the extraction parameters are stable, and the extraction process is feasible.

[0371] 11.2 Concentration process

[0372] Table 17: Concentration process diagram

[0373]

[0374] From the above results, it can be seen that the concentration process is smooth, the concentration process parameters are stable, and the concentration method is feasible.

[0375] 11.3 Drying process

[0376] Table 18: Drying process table

[0377]

[0378] From the above results, it can be seen that the drying process is smooth, the spray drying yield is about 95%, the spray drying process parameters are stable, and the drying method is feasible.

[0379] 11.4 Preparation process

[0380] Table 19: Preparation process table

[0381]

[0382]

[0383] From the above results, it can be seen that the granulation process of the preparation is smooth, the granule yield is about 90%, the granulation process parameters are stable, and the granulation method is feasible.

[0384] 12. Process research summary

[0385] (1) The extraction, concentration, drying, and formulation processes of the three pilot production batches were stable and the method was feasible;

[0386] (2) The three batches of pilot production of the Chinese medicine composition granules of the present invention were stable, and the quality of the finished products met the quality standard requirements. The production and preparation process of the Chinese medicine composition granules of the present invention is feasible.

[0387] Example 6: Preparation method of Chinese medicine composition involving quality control process

[0388] In summary, if Figure 20 As shown, the preparation process of the Chinese medicine composition of the present invention is determined as follows:

[0389] Add water and boil twice, each time for 1 hour, filter, combine the decoctions, and concentrate the filtrate under reduced pressure to a clear paste with a relative density of 1.03-1.05 (70°C). Add appropriate amount of maltodextrin, dry, or crush to make 1000g of granules.

[0390] In a specific embodiment, 10 times the amount of water is added for the first time, soaked for 0.5 hours, and boiled for 1.0 hour; 10 times the amount of water is added for the second time, boiled for 1.0 hour, the decoctions are combined, and filtered.

[0391] In any of the above specific embodiments, the filtrate is concentrated under reduced pressure at 75°C ± 5°C to a clear paste with a density of 1.03 to 1.05 (70°C), and centrifuged at 10,000 rpm / min.

[0392] In any of the above specific embodiments, take the above centrifuged clear paste, add an appropriate amount of maltodextrin (the total amount of each prescription is 40g, and the solid content of the centrifuged clear paste is about 10.5% of the prescription amount), dissolve, mix, and spray dry to obtain extract powder.

[0393] In any of the above specific embodiments, extract powder is taken, an appropriate amount of binder (water) is added, granulated by boiling, and granulated to prepare 1000 g of granules.

[0394] In any of the above specific embodiments, the particles are packaged into 10 g / bag and packed with composite film.

[0395] After the granulation process is completed, samples are randomly selected and thin layer chromatography is performed on Scutellaria baicalensis, Glycyrrhiza uralensis and Coptis chinensis according to the developing agent combination determined in Examples 1-3.

[0396] Based on the identification results, determine whether the product meets the quality standards of the Chinese Pharmacopoeia. If the quality is determined to be qualified according to the thin layer chromatography results, proceed to the subsequent processes such as total mixing. If the quality is determined to be unqualified according to the thin layer chromatography results, stop production. Check all raw materials and production processes. For the full process, see Figure 20 .

[0397] Example 7: Applicability test of the microbiological examination method of the prepared Chinese medicine composition of the present invention

[0398] 1. Materials and equipment

[0399] 1.1 Test materials and strains

[0400] (1) Test materials

[0401] Trypticase soytone agar (lot number 20171103), trypticase soytone broth (lot number 20171202), Sabouraud dextrose agar (lot number 20171207), MacConkey broth (lot number 20171123), MacConkey agar (lot number 20170810), and pH 7.0 peptone-sodium chloride buffer (lot number 20171121) were all provided by Qingdao Hi-Tech Industrial Park Haibo Biotechnology Co., Ltd. Sodium chloride (lot number 20170121, Sinopharm Chemical Reagent Co., Ltd.) was also provided.

[0402] (2) Bacterial strains

[0403] Table 20 Bacteria species

[0404]

[0405] 1.2 Instruments and Equipment

[0406] Biochemical incubator (LRH-70, Shanghai Yiheng Scientific Instrument Co., Ltd.); water-proof constant temperature incubator (Shanghai Yiheng Scientific Instrument Co., Ltd.); water bath (HWS24, Shanghai Yiheng Scientific Instrument Co., Ltd.); electronic balance (YP20002, Shanghai Youke Instrument Co., Ltd.); precision blast drying oven (BPG-9140A, Shanghai Yiheng Scientific Instrument Co., Ltd.); high pressure sterilizer (G154TW Zhiwei (Xiamen) Instrument Co., Ltd.).

[0407] 2. Methods and Results

[0408] 2.1 Preparation of bacterial suspension

[0409] 2.1.1 Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Escherichia coli

[0410] The tryptic soy culture cultured at 33°C for 18 hours was diluted 10-fold with sterile 0.9% sodium chloride solution to prepare a bacterial suspension containing no more than 10,000 cfu / ml for later use.

[0411] 2.1.2 Candida albicans

[0412] The Sabouraud glucose culture cultured at 25°C for 48 hours was diluted with sterile 0.9% sodium chloride solution in 10-fold increments to prepare a bacterial suspension containing no more than 10,000 cfu / ml for later use.

[0413] 2.1.3 Aspergillus niger

[0414] To a Sabouraud dextrose agar slant culture incubated at 25°C for 10 days, add 5 ml of a sterile 0.9% sodium chloride solution containing 0.05% polysorbate 80 to elute Aspergillus niger spores. Pipette the bacterial suspension into a sterile test tube and dilute it in 10-fold increments with a sterile 0.9% sodium chloride solution containing 0.05% polysorbate 80 to prepare a bacterial suspension containing no more than 10,000 cfu / ml. Set aside.

[0415] 2.2 Preparation of test solution

[0416] Weigh about 10 g of the granules of the Chinese medicine composition of the present invention (batch numbers: 20190901, 20190902, 20190903), add pH 7.0 sterile sodium chloride-peptone buffer to 100 ml, mix evenly in a 45 ° C water bath to prepare a 1:10 test solution.

[0417] 2.3 Verification test for total aerobic bacteria, molds and yeast counts

[0418] 2.3.1 Test method (conventional method)

[0419] ① Test group: Take 0.1ml of Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans, and Aspergillus niger suspension and 9.9ml of test solution respectively into a 50ml test tube and mix well, then take 1ml of the mixed liquid and inject it into a plate, immediately pour it into tryptic soy peptone agar medium for culture, prepare 2 plates in parallel for each test bacteria, place them in a constant temperature incubator at 30-35℃ for 3-5 days, observe and record the number of colonies. In addition, take 0.1ml of Candida albicans suspension, Aspergillus niger suspension and 9.9ml of test solution respectively into a 50ml test tube and mix well, then take 1ml and inject it into a plate, immediately pour it into Sabouraud dextrose agar medium for culture, prepare 2 plates in parallel for each test bacteria, place them in a constant temperature incubator at 20-25℃ for 5-7 days, observe and record the number of colonies;

[0420] ② Bacterial liquid group: Take 0.1ml of each test bacterial suspension and 9.9ml of pH 7.0 sodium chloride peptone buffer into a 50ml test tube and mix well. Then take 1ml and inject it into a plate. Pour it into the corresponding agar medium for culture. Prepare two parallel plates for each test bacterial strain and culture them under the corresponding conditions. Observe and record the number of colonies.

[0421] ③ Test sample control group: Take 9.9 ml of the test solution and 0.1 ml of pH 7.0 sodium chloride peptone buffer solution into a 50 ml test tube and mix well. Then take 1 ml and inject it into a plate. Pour it into the corresponding agar medium for culture. Prepare 2 plates in parallel for each batch of test samples, culture them under the corresponding conditions, observe and record the number of colonies.

[0422] 2.3.2 Test results

[0423] Calculate the recovery rate using the following formula; the result should be within the range of 50% to 200%. The total aerobic bacteria count, mold count, and yeast count for this product can be tested using the conventional method (Chinese Pharmacopoeia 2015 Edition, Part IV).

[0424] The recovery rate of the experimental group = [(average colony count of the experimental group - average colony count of the test sample control group) / average colony count of the bacterial solution group] × 100%.

[0425] Table 21 Verification results of the applicability of the method for counting aerobic bacteria, molds and yeast colonies in the granules of the Chinese medicine composition of the present invention

[0426]

[0427]

[0428] Results: As can be seen from the above table, the plate method can be used to count the microorganisms of the granules of the Chinese medicine composition of the present invention. In the experiment, the colony recovery rates of Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans and Aspergillus niger were all in the range of 50% to 200%, indicating that the plate method is feasible for counting aerobic bacteria, molds and yeasts in the granules of the Chinese medicine composition of the present invention.

[0429] 2.4 Validation of control bacteria test methods

[0430] 2.4.1 Test method

[0431] ① Experimental group: 10 ml of the test solution was inoculated into 100 ml of trypticase soy broth, and a suspension of Escherichia coli with a colony count of no more than 100 cfu was added. The culture was incubated at 33°C for 24 h. 1 ml of the culture was inoculated into 100 ml of MacConkey broth and incubated at 44°C for 24 h. The culture was then streaked onto MacConkey agar and incubated at 35°C for 24 h.

[0432] ② Test Group: Inoculate 10 ml of the test solution into 100 ml of trypticase soytone broth. Incubation conditions and examination methods are the same as those for the experimental group. ③ Negative Control Group: Add 10 ml of sterile pH 7.0 sodium chloride-peptone buffer to 100 ml of trypticase soytone broth. Incubation conditions and examination methods are the same as those for the experimental group.

[0433] 2.4.2 Test results

[0434] The table below shows that the test group tested positive, with typical bright pink colonies. The colonies on the plate were isolated, cultured, and Gram-stained, confirming them as Escherichia coli. Both the test sample group and the negative control group tested negative. Therefore, conventional methods can be used to test for control bacteria in this product.

[0435] Table 22 Verification results of the applicability of the control method of bacteria in the Chinese medicine composition granules of the present invention

[0436] batch number experimental group Test sample group Negative control group 20190901 + - - 20190902 + - - 20190903 + - -

[0437] Note: Escherichia coli colony count: 63 / 57

[0438] 3. Experimental Results

[0439] 3.1 According to Table 2, the conventional method was used to check the recovery rates of three batches of the granules of the Chinese medicine composition of the present invention against five test bacteria, namely Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans (TSA culture), Candida albicans (SDA culture), Aspergillus niger (TSA culture), and Aspergillus niger (SDA culture), all of which were within the range of 50% to 200%. Therefore, this product has no antibacterial effect.

[0440] 3.2 According to Table 3, when the conventional method was used to conduct the methodological verification test and inspection of control bacteria, the positive bacteria grew well, indicating that this product had no antibacterial effect on Escherichia coli.

[0441] 3.3 In summary, this product can be tested for microbial limits and control bacteria using conventional methods.

[0442] Example 8: Clinical testing of Chinese medicine compositions

[0443] 1. Prepare the following proportions of the prescription pieces:

[0444] Test group 1: 19 parts of Forsythia suspensa, 16 parts of Pinellia ternata, 11 parts of Radix Glycyrrhizae Preparata, 11 parts of Radix Bupleuri, 9 parts of Codonopsis pilosula, 9 parts of Dry Ginger, 7 parts of Radix Scutellariae Baicalensis, 6 parts of Coptis chinensis, 7 parts of Rhubarb, and 22 parts of Jujube.

[0445] Test group 2: 21 parts of Forsythia suspensa, 14 parts of Pinellia ternata, 13 parts of Radix Glycyrrhizae Preparata, 13 parts of Radix Bupleuri, 11 parts of Codonopsis pilosula, 7 parts of Dry Ginger, 5 parts of Radix Scutellariae, 4 parts of Rhizoma Corydalis, 5 parts of Rhubarb, and 18 parts of Jujube.

[0446] Test group 3: 20 parts of Forsythia suspensa, 15 parts of Pinellia ternata, 12 parts of Radix Glycyrrhizae, 12 parts of Radix Bupleuri, 10 parts of Codonopsis pilosula, 8 parts of Dry Ginger, 6 parts of Radix Scutellariae, 5 parts of Coptis chinensis, 6 parts of Rhubarb, and 20 parts of Jujube.

[0447] Control group 1: commercially available watermelon frost spray (Guilin Sanjin Pharmaceutical Co., Ltd.).

[0448] Control group 2: commercially available Sanhuang tablets (Zhongjing Wanxi Pharmaceutical Co., Ltd.).

[0449] 2. Patient samples

[0450] The 276 patients selected were all treated at the National Medical Hall of Nanjing University of Chinese Medicine and met the diagnostic criteria for chronic oral ulcers. The diagnosis was based on the following:

[0451] (1) Oral ulcers that have not healed for more than 10 days, or long-term recurrent oral ulcers;

[0452] (2) Red tongue with teeth marks on the edges, greasy coating with white saliva, and a smooth and thready pulse; and / or;

[0453] (3) Long-term abdominal distension, abdominal pain or belching; and / or, accompanied by constipation or sensitive diarrhea.

[0454] 3. Treatment methods

[0455] Take the granules with water twice daily for a 7-day course of treatment. Avoid eating raw, cold, spicy, irritating, or rotten foods during this period. For the spray or tablets, administer according to the instructions. A 7-day course of treatment is considered a complete course.

[0456] 4. Efficacy standards

[0457] Markedly effective: After one course of treatment, there is no mucosal congestion or redness around the oral ulcer, the ulcer wound disappears, and / or the accompanying abdominal distension, abdominal pain or constipation, easy diarrhea and other symptoms disappear; there is no recurrence for more than 3 months.

[0458] Effective: After one course of treatment, the size and number of oral ulcers are reduced, there is a red halo around them, and the pain is significantly relieved; recurrence is delayed for more than 3 months, and the symptoms are significantly alleviated after recurrence, and / or, occasionally accompanied by abdominal distension, abdominal pain or constipation, easy diarrhea and other symptoms.

[0459] Basically effective: oral ulcers are reduced in size and pain is relieved after one course of treatment, but relapse occurs within 10 days after discontinuation of medication, and / or symptoms such as abdominal distension, abdominal pain, constipation, and easy diarrhea are not significantly improved.

[0460] Completely ineffective: After one course of treatment, the mucosa around the oral ulcer becomes congested or red, the ulcer wound surface does not change, the pain worsens or does not significantly improve, and / or symptoms such as abdominal distension, abdominal pain or constipation, and easy diarrhea worsen or do not significantly improve.

[0461] 5. Clinical Results

[0462] Several examples that met the above criteria were selected from each embodiment, and the test results after 7 days are shown in Table 23:

[0463] Table 23: Comparison of Effectiveness

[0464] experimental group Case Significantly effective efficient Basically effective Completely ineffective Efficiency Test Group 1 49 32 10 7 0 85.7％ Test Group 2 65 50 8 7 0 89.2％ Test Group 3 87 69 9 9 0 89.7％ Control group 1 40 4 6 9 21 25.0％ Control group 2 35 9 16 8 2 71.4％

[0465] Total effective rate = (markedly effective + effective) / total number of cases × 100%. Since the basically effective group is prone to relapse, it is not included in the statistics.

[0466] After analysis of clinical trial data, the efficacy of the drug of the present invention reached 85.7%-89.7%. Compared with the control group, the present invention can not only quickly treat recurrent oral ulcers, but also significantly improve symptoms such as abdominal distension, abdominal pain or constipation, and susceptible diarrhea.

[0467] 6. Specific case description

[0468] Patient Yuan had suffered from oral ulcers for 10 years and gallbladder polyps for 5 years, all related to chronic stress. He also suffered from mild insomnia, susceptibility to cold, and abdominal distension. His tongue was red with a greasy coating, and his pulse was thready. He took the granules in Test Group 3 with boiled water twice daily for a course of 7 days. During the medication period, he was advised to avoid raw, cold, spicy, irritating, or rotten foods. After two courses of treatment, his oral ulcers healed and his abdominal distension resolved. There was no recurrence six months later.

[0469] Patient Zhu had suffered from oral ulcers and constipation for many years, with scattered ulcers in her mouth. She also experienced a bitter taste in her mouth, a cough, poor appetite, a red tongue with a greasy coating, and a thready pulse. She took the granules in Test Group 3 with boiled water twice daily for a seven-day course of treatment. After two courses, her oral ulcers healed and her bowel movements returned to normal. Two years later, she sought medical attention due to menopause, and the results were satisfactory.

[0470] Patient Yang had been experiencing recurrent oral ulcers for three years, benign prostatic hyperplasia, and cerebral infarction for three years. He had a red and swollen nasal cavity, a sticky mouth, a bitter taste in the morning, and a burning sensation. He also had a tingling sensation in his mouth, a bloating in his stomach, and dry, hard stools every two days, like goat dung. He also had palpitations, weakness in his lower limbs, and incomplete urination. He also had occasional calf cramps. His tongue was red and had a greasy coating, and his pulse was weak and rapid. He took the granules in Test Group 3 with boiled water twice daily for seven days. After three courses of treatment, his oral ulcers healed, his bowel movements became normal, and his bloating disappeared.

[0471] Patient Xu suffered an allergic asthma attack during the Lunar New Year holiday, resulting in oral ulcers, cough, and left submandibular lymphadenopathy. He developed a fever after scraping. He also spoke rapidly and moved briskly. He took the granules in Test Group 2 with boiled water twice daily for a course of seven days. The oral ulcer healed after seven days, and after another course of treatment, there was no recurrence within a year.

[0472] Patient Wang suffered from recurrent oral ulcers and had long-term success with Aoke for gastritis. He also experienced fatigue, dull stomach pain, aggravated by cold, a red tip of the tongue, a greasy yellow coating, and a stringy pulse. He took the granules from Test Group 3 with water twice daily for a seven-day course. After one course, his oral ulcers healed, his restlessness and pressure were alleviated, and his red tongue, greasy coating, and stringy pulse disappeared. He continued taking the granules for another course, and his gastritis subsided and the pain disappeared.

[0473] Patient Zhu had recurring oral ulcers for 2-3 years. His bowel movements and appetite were normal, but he had bloating, a red tongue with teeth marks, and a greasy coating with white saliva. He took the granules in Test Group 3 with boiled water twice daily for a course of 7 days. After two courses, his oral ulcers healed and his bloating disappeared. He experienced no recurrence within six months.

Claims

1. A method for preparing a Chinese medicine composition based on thin layer chromatography identification, characterized in that: The following steps are involved: (1) Weigh the following Chinese medicinal ingredients in parts by weight: 19-21 parts of old forsythia root, 14-16 parts of ginger pinellia root, 11-13 parts of roasted liquorice root, 11-13 parts of bupleurum root, 9-11 parts of codonopsis pilosula, 7-9 parts of dried ginger, 5-7 parts of kushen, 4-6 parts of yalian, 5-7 parts of cooked rhubarb, and 18-22 parts of jujube; (2) Decoction the above Chinese herbal components with water twice, each time for 0.5-1.0 hours, filter, and combine the decoctions; (3) At 60°C, the filtrate was concentrated under reduced pressure to a clear paste with a relative density of 1.03 to 1.10; (4) adding an appropriate amount of matrix to the clear paste obtained in step (3), stirring evenly, and drying to granulate to obtain a clear paste powder; (5) Conduct quality inspection on the dried paste powder and guide the standardized production of the product based on the quality inspection results; (6) Based on the quality test results, different disintegrants, fillers, binders, lubricants and / or bases are added to the dry extract powder that meets the quality standards to prepare granules, tablets, capsules, pills or powders; The quality inspection step in step (5) is: ① Take the Chinese medicine composition granules, add water to dissolve, add n-butanol to extract, and combine the extracts; ② Evaporate the extract to dryness in a water bath and dissolve the residue in methanol as the test solution; prepare positive and negative control solutions in parallel using the same method; ③ Pipette the test solution, positive control solution, and negative control solution separately, place them on the same silica gel G thin layer plate, develop them on the thin layer identification developer, and take them out; ④ Add color developing solution, heat with hot air until the spots are clearly colored, and inspect under ultraviolet light; ⑤ Compare the spots of the test sample, positive control, and negative control. If the test sample and the positive control have spots of the same color at the corresponding positions, and there are no interfering spots in the negative control, the test sample is judged to contain the drug component with the same or similar quality as the positive control. For Forsythia suspensa, chloroform and methanol were mixed in a volume ratio of 8:1 as the developing solvent; For Coptis chinensis, a mixture of toluene, ethyl acetate, methanol, isopropanol, and water in a volume ratio of 6:3:2:1.5:0.3 was used as a developing agent and placed in a developing cylinder pre-saturated with concentrated ammonia solution for 20 minutes. For licorice, ethyl acetate: glacial acetic acid: water was mixed in a volume ratio of 15:2:2 as the developing solvent.

2. The method for preparing a Chinese medicine composition based on thin layer chromatography identification according to claim 1, characterized in that: For Forsythia suspensa, add n-butanol to extract twice in steps ① and ②, combine the extracts, evaporate to dryness in a water bath, and dissolve the residue in methanol as the test solution; Take 0.2 g of the positive control medicinal material, add appropriate amount of water, reflux extraction for 60 minutes, cool, filter, and concentrate the filtrate to 10 ml. Take 5 ml of the negative sample solution without Forsythia suspensa, extract the two with n-butanol twice, 15 ml each time, combine the extracts, evaporate to dryness in a water bath, and dissolve the residue in 2 ml of methanol as the control medicinal material solution; Step ③ is to draw 6 μl of the test solution and 4 μl of the control medicinal material solution, respectively spot them on the same silica gel G thin layer plate, and use chloroform and methanol mixed in a volume ratio of 8:1 as a developing solvent for development; In step ④, the surface is sprayed with 10% sulfuric acid ethanol solution and dried at 105° C. until the spots are clear.

3. The method for preparing a Chinese medicine composition based on thin layer chromatography identification according to claim 1, characterized in that: For Coptis chinensis, steps ① and ② were extracted twice with n-butanol, the extracts were combined, evaporated to dryness in a water bath, and the residue was dissolved in methanol as the test solution; Step ③ is to draw 3 μl of the test solution and 2 μl of the Rhizoma Coptidis control medicinal material solution, respectively, onto the same silica gel G thin layer plate, and adjust the volume ratio of toluene: ethyl acetate: methanol: isopropanol: water to 6:3:2:1.5:0.3 as a developing solvent, and place it in a developing cylinder pre-saturated with concentrated ammonia test solution for 20 minutes for development; In step ④, place the sample in a developing cylinder that has been pre-saturated with concentrated ammonia solution for 20 minutes, develop the sample, take it out, dry it, and examine it under a 365nm ultraviolet lamp.

4. The method for preparing a Chinese medicine composition based on thin layer chromatography identification according to claim 3, characterized in that: The step ① is to take 0.5g of the finished granules, add 5ml of water to dissolve, add n-butanol to extract twice, 10ml each time, combine the extracts, evaporate to dryness in a water bath, and dissolve the residue in 2ml of methanol as the test solution; In step ②, 0.25 g of each of the positive and negative control medicinal materials were taken, n-butanol was added, ultrasonic treatment was performed for 30 min, the mixture was filtered, the filtrate was evaporated to dryness, and the residue was dissolved in 2 ml of methanol as the control medicinal material solution.

5. The method for preparing a Chinese medicine composition based on thin layer chromatography identification according to claim 1, characterized in that: For licorice, add n-butanol to steps ① and ② and extract twice, combine the extracts, evaporate to dryness in a water bath, and dissolve the residue in methanol as the test solution; In step ③, 7 μl of the test solution and 3 μl of the control medicinal material solution were respectively aspirated and spotted on the same silica gel G thin layer plate, and a mixture of ethyl acetate: glacial acetic acid: water in a volume ratio of 15:2:2 was used as a developing solvent for development; In step ④, the sample was sprayed with 10% sulfuric acid ethanol solution, heated at 105° C. until the spots were clear, and then inspected under a 365 nm ultraviolet lamp.

6. The method for preparing a Chinese medicine composition based on thin layer chromatography identification according to claim 5, characterized in that: In step ②, 0.1 g of the positive liquorice was taken, an appropriate amount of water was added, and reflux extraction was performed for 60 minutes. The mixture was cooled, filtered, and the filtrate was concentrated to 20 ml. The mixture was extracted twice with n-butanol, 15 ml each time. The extracts were combined and evaporated to dryness in a water bath. The residue was dissolved in 2 ml of methanol as a control medicinal material solution.

7. The method for preparing a Chinese medicine composition based on thin layer chromatography identification according to any one of claims 1 to 6, characterized in that: In step (2), add 8-10 times the amount of water for the first time, soak for 0.5 hours, and boil for 1.0 hours; add 6-8 times the amount of water for the second time, boil for 0.5 hours, combine the decoctions, and filter; In the step (3), the filtrate obtained in the step (2) is concentrated under reduced pressure at 60° C. to obtain a clear paste with a density of 1.03 to 1.05, and the clear paste is centrifuged at 10,000 rpm / min; In the step (4), the above-mentioned centrifuged clear paste is taken, an appropriate amount of maltodextrin is added, dissolved, mixed, and spray-dried to obtain a dry extract powder; The quality inspection in step (5) includes randomly selecting granulated samples, performing thin layer chromatography identification using the aforementioned developer set, inspecting and evaluating the quality of the product, and judging whether the product quality is qualified, thereby determining whether to continue or interrupt the production process of the product.

8. The method for preparing a Chinese medicine composition based on thin layer chromatography identification according to claim 7, characterized in that: In step (6), the disintegrant includes one or more of starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, cross-linked polyvinyl pyrrolidone, low-substituted hydroxypropyl cellulose, and cross-linked sodium carboxymethyl cellulose; The filler includes one or more of starch, mannitol, chitin, pregelatinized starch, lactose, microcrystalline cellulose, and sucrose; The lubricant includes one or more of sodium lauryl sulfate, magnesium stearate, talc, and silicon dioxide; The adhesive comprises one or more of starch slurry, hydroxypropyl methylcellulose and polyvinyl pyrrolidone; The matrix includes maltodextrin or one of PEG4000 and PEG6000.

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