Traditional Chinese medicine preparation with throat clearing and nourishing functions as well as preparation method and application of traditional Chinese medicine preparation

Abstract

The invention relates to the technical field of throat health care products, in particular to a traditional Chinese medicine preparation with throat clearing and nourishing functions and a preparation method and application thereof. The composition is composed of honeysuckle, radix scrophulariae, Chinese olive and other medicines, animal tests and human body tests prove that the product is safe and non-toxic, achieves a good effect in the aspect of clearing and nourishing the throat, and is suitable for popularization and application.

Description

Technical Field

[0001] The invention relates to a traditional Chinese medicine preparation with the function of clearing the throat and moistening the throat, a preparation method and an application thereof. The components contained in the composition are all varieties listed in the catalogue of traditional Chinese medicines with medicinal and edible properties, and the composition belongs to the field related to food and health products. Background Art

[0002] Chronic pharyngitis is one of the most common diseases in otolaryngology. It refers to diffuse pharyngeal lesions caused by chronic infection. It mostly occurs in adults and is often accompanied by other upper respiratory tract diseases. Chronic pharyngitis is often caused by repeated attacks of acute pharyngitis, pus from rhinitis and sinusitis irritating the pharynx, or nasal congestion and mouth breathing.

[0003] Generally, there are no obvious systemic symptoms. The main clinical manifestations are pharyngeal discomfort, dryness, a foreign body sensation or mild pain, a dry cough, nausea, dark red pharyngeal congestion, and visible lymphoid follicles on the posterior pharyngeal wall. Patients with chronic pharyngitis often clear their throats and produce white sputum due to increased pharyngeal secretions.

[0004] Chronic pharyngitis often has a history of recurrent acute pharyngitis, or may be caused by long-term mouth breathing due to nasal disease, excessive smoking and drinking, dry air, dust and irritant gas pollution, etc. Symptoms include: discomfort in the throat, or pain, itching, dryness, burning, smoky sensation, foreign body sensation, etc.; irritating cough, forceful coughing up secretions in the morning, or even nausea. The course of the disease is more than 2 months, and symptoms are often caused by colds, fatigue, and excessive talking. Examination can reveal chronic and worsening congestion in the pharynx. It is dark red or has dendritic congestion; there is hyperplasia of the lymphoid follicles on the posterior pharyngeal wall, or enlargement of the lateral pharyngeal cords; hyperplasia and hypertrophy of the pharyngeal mucosa, or dryness, atrophy, thinning, and adherence of secretions.

[0005] Western medicine generally focuses on treating the cause and applying topical medications. For example, mouthwashes such as gargling with a 2% boric acid solution, 3% saline, and a 1:5000 nitrofurazone solution, or applying a 2% iodine glycerol or 5% strong protein silver solution to the pharyngeal wall, or oral administration of iodine lozenges, domiphene, or mint, can alleviate symptoms to some extent. Local symptomatic treatment can also include traditional Chinese medicines such as coral lozenges and watermelon frost. Quitting smoking and alcohol is crucial.

[0006] Traditional Chinese Medicine (TCM) treatment of this disease focuses on addressing the root cause, using medications based on syndrome differentiation, resulting in good efficacy. The general medications are: For Yin deficiency and fire inflammation, use Yangyin Qingfei Decoction (with ingredients like Ophiopogon japonicus, Radix Rehmanniae, Scrophulariae ningpoensis, White Peony Root, Licorice Root, Gypsum Fibrosum, Menthol, Fritillaria cirrhosa Powder, Cortex Moutan, and Morus alba); for phlegm-blocking and blood stasis, use Xiaolu Pills (with ingredients like Scrophulariae ningpoensis, Oyster, Fritillaria cirrhosa, Radix Rehmanniae, Ophiopogon japonicus, Trillium, Laminaria japonica, and Seaweed); for Yin deficiency and fluid depletion, use Shengmai Yin (with ingredients like White Sun-dried Ginseng, Ophiopogon japonicus, Schisandra chinensis, Dendrobium officinale, Polygonatum odoratum, Imperata cylindrica, and Rhizoma Anemarrhenae). Take daily, frequently, to moisten the throat.

[0007] Chronic pharyngitis is a difficult disease to treat and has a long course. Long-term use of medication, whether Western or traditional Chinese medicine, can cause toxic side effects. Therefore, the market urgently needs a long-term health food with throat-clearing and soothing effects for chronic pharyngitis patients.

[0008] The present invention provides a traditional Chinese medicine preparation having the health-care functions of clearing the throat and moistening the throat, in particular to the use of the preparation in preparing medicines or health-care foods for treating or improving chronic pharyngitis. Summary of the Invention

[0009] The present invention aims to provide a traditional Chinese medicine preparation with the function of clearing the throat and moistening the throat.

[0010] The traditional Chinese medicine preparation of the present invention is prepared from the following raw materials in parts by weight: 36 parts of honeysuckle; 36 parts of Scrophularia ningpoensis; 30 parts of green fruit; 18 parts of Platycodon grandiflorum; 18 parts of Emblica officinalis; 3 parts of Momordica grosvenori; 0.12 parts of menthol; 30 parts of lactose; 16.4 parts of dextrin; 6 parts of erythritol; 0.4 parts of magnesium stearate; and 0.04 parts of sucralose. The dosage form of the traditional Chinese medicine preparation is tablets, and each 100 grams of the tablets contains a total amount of not less than 400 mg of total saponins and not less than 240 mg of chlorogenic acid.

[0011] Furthermore, the total saponins are determined by UV-visible spectrophotometry, comprising the following steps:

[0012] 1) Preparation of standard solution:

[0013] Weigh 0.020 g of ginsenoside Re standard, dissolve it in methanol and dilute to 10.0 mL, i.e., 2.0 mg of ginsenoside Re per mL;

[0014] 2) Preparation of test solution:

[0015] Take 10 tablets, grind them into powder, take 1.5 g, place them in a 100 mL volumetric flask, add 70 mL of water, sonicate for 40 minutes, then dilute to 100 mL with water, let stand, and the supernatant is used as the test solution;

[0016] 3) Column chromatography:

[0017] Use a glass chromatography column filled with 3 cm of Amberlite-XAD-2 macroporous resin and topped with 1 cm of neutral alumina. Wash the column with 25 mL of 70% ethanol, discard the eluent, then wash the column with 25 mL of water, discard the eluent, add 1.0 mL of the test solution, wash the column with 25 mL of water, discard the eluent, and elute the ginsenosides with 25 mL of 70% ethanol. Collect the eluent in an evaporating dish and evaporate to dryness in a 60°C water bath.

[0018] 2) Color reaction:

[0019] Add 0.2 mL of 5% vanillin glacial acetic acid solution to the dried evaporating dish to dissolve the residue. Then add 0.8 mL of perchloric acid and transfer to a 5 mL graduated centrifuge tube with a stopper. Heat in a 60°C water bath for 10 minutes. After cooling in an ice bath, add 5.0 mL of glacial acetic acid to obtain the product.

[0020] 3) Determination

[0021] The same method as steps 3) and 4) is used for column chromatography and color reaction of the standard solution. The absorbance is measured at 560 nm by UV-visible spectrophotometry to calculate the total saponin content in the test sample.

[0022] Furthermore, the chlorogenic acid was determined by high performance liquid chromatography, and the liquid chromatography conditions were as follows:

[0023] Chromatographic column: ODS C18 column, 250 mm × 4.6 mm, 5 μm;

[0024] Mobile phase: 0.5% acetic acid solution: acetonitrile = 9:1 (V / V);

[0025] Flow rate: 1.0 mL / min;

[0026] Column temperature: 35°C;

[0027] Detection wavelength: 327nm;

[0028] Injection volume: 10 μL.

[0029] Furthermore, in the determination of chlorogenic acid by high performance liquid chromatography, the following steps are also included:

[0030] Preparation of chlorogenic acid standard solution: Weigh 0.02 g of chlorogenic acid standard into a 100.0 mL volumetric flask, dissolve it with mobile phase and make up to volume, then shake well, i.e., each mL contains 0.2 mg of chlorogenic acid;

[0031] Preparation of sample solution: Take 10 tablets, grind them into powder, weigh 0.2 g, add 20 mL of 70% methanol to a 25.0 mL volumetric flask, sonicate for 30 minutes, dilute to volume with 70% methanol, filter through a 0.45 μm microporous membrane, and use the filtrate as the sample solution;

[0032] Preparation of standard curve: Pipette chlorogenic acid standard solution, dilute with mobile phase and prepare a series of standard solutions with concentrations of 2.00, 10.0, 20.0, 40.0, and 80.0 μg / mL in a volumetric flask for HPLC determination. Draw a standard curve with the measured chromatographic peak area as the ordinate and the corresponding standard solution concentration as the abscissa.

[0033] Sample determination: inject the standard solution and sample solution into the liquid chromatograph, determine, and calculate the chlorogenic acid content in the sample.

[0034] Furthermore, the preparation method of the tablet having the function of clearing the throat and moistening the throat comprises the following steps:

[0035] (1) Menthol grinding: Weigh menthol, grind it, pass it through a 100-mesh sieve, and set aside;

[0036] (2) Extraction: Take honeysuckle, Scrophularia, Cibotrys, Platycodon, Emblica, and Momordica grosvenori according to the formula ratio, add water and extract twice, each time for 1 hour, add 12 times the amount of water for the first time and 10 times the amount of water for the second time, filter the extract and set aside:

[0037] (3) Concentration: The extract is concentrated under reduced pressure at a temperature of 60-80°C and a vacuum degree of 0.04-0.08 MPa to form a clear paste with a relative density of 1.08-1.12; an appropriate amount of ethanol is added to the clear paste to make the alcohol content 70%, the mixture is stirred evenly, and the mixture is allowed to stand at 2-10°C for 48 hours; the mixture is filtered, the filtrate is collected, the ethanol is recovered under reduced pressure, and the mixture is concentrated at a temperature of 60-80°C and a vacuum degree of 0.04-0.08 MPa to form a thick paste with a relative density of 1.20-1.30;

[0038] (4) Drying: Use vacuum belt drying with a heating plate temperature of 110-120°C and a vacuum degree of 0.08-0.10 MPa to collect the dry paste for later use;

[0039] (5) Grinding: Grind the dry paste obtained by vacuum belt drying, pass it through a 100-mesh sieve, and set aside;

[0040] (6) Granulation: Place the above dry powder in a mixer, add lactose, dextrin and erythritol, mix well, add 80-90% ethanol to make a soft material, granulate with a 16-mesh sieve, and dry at below 60°C.

[0041] (7) Granulation: Take the dried granules and place them in a crusher, and sieve them through a 1.5 mm conical sieve;

[0042] (8) Total mixing: Add menthol powder, magnesium stearate, and sucralose to the granules after granulation and mix them evenly in a mixer at 8 rpm for 20 minutes;

[0043] (9) Tablet pressing: Tablets are pressed into tablets, and each tablet weighs 0.8 g.

[0044] The present invention also provides the use of the above-mentioned traditional Chinese medicine preparation with the function of clearing the throat and moistening the throat in preparing the throat-clearing and moistening medicine or health food.

[0045] Furthermore, the medicine or health food is a medicine or health food for treating or improving chronic pharyngitis.

[0046] Chronic pharyngitis is a chronic disease characterized by dry, burning pain in the throat, a foreign body sensation, discomfort, itchy throat with scant pharyngitis, and a dry cough. Clinically, it is categorized as chronic simple pharyngitis, chronic hypertrophic pharyngitis, and atrophic or dry pharyngitis. Patients often experience discomfort in the throat, a foreign body sensation, or an irritating cough in the morning, particularly when brushing their teeth, often accompanied by nausea. Chronic pharyngitis falls under the category of "throat bi" in traditional Chinese medicine. The term "throat bi" was first mentioned in the Yellow Emperor's Classic of Internal Medicine, my country's first medical treatise. "Suwen: Discussion on Yin and Yang" states: "When one yin and one yang clash, it causes throat bi." The onset of chronic pharyngitis is often due to visceral yin deficiency. Lung yin deficiency prevents the upward transport of body fluids, leading to internal heat and a burning sensation in the throat. Kidney yin deficiency, with insufficient body fluids, deprives the throat of nourishment, causing internal heat to accumulate along the meridians and rise to the throat. Yin deficiency leads to insufficient body fluids, which deprive the throat of nourishment. This leads to the accumulation of deficient fire along the meridians in the throat. Alternatively, wind-heat attacks the lungs and stomach, triggering stagnant heat, which rises and stagnates in the throat, causing the disease. Furthermore, liver dysfunction leads to upward flow of qi, causing stagnation in the throat. Splenic dysfunction leads to internal retention of dampness, which in turn generates phlegm, which accumulates in the throat. Fire, qi, and phlegm interfere with each other, leading to prolonged blood stasis in the throat. Therefore, the pathogenesis of this condition is primarily due to excessive fire caused by yin deficiency, but may also be accompanied by the accumulation of qi stagnation, phlegm coagulation, and blood stasis, leading to localized accumulation of pathological products in the throat.

[0047] In the Chinese medicine preparation of the present invention, the Latin names of the raw materials used as active ingredients and their processing methods are from the "Chinese Materia Medica Dictionary" (first edition, July 1977, Shanghai Science and Technology Press) and the "Chinese Pharmacopoeia" (2005 edition, Chemical Industry Press).

[0048] Honeysuckle: Sweet and cold, it enters the lung, heart, and stomach meridians, clearing away heat and detoxifying, and dispersing wind-heat. It was first mentioned in Tao Hongjing's "Ming Yi Bie Lu," which states that it "treats cold and heat, swelling, and long-term use of it can reduce weight and prolong life." Tang Shenwei's "Zheng Lei Ben Cao" from the Northern Song Dynasty also states that it "treats cold and heat, swelling, and long-term use can reduce weight and prolong life." "Materia Medica" says that it "tastes spicy, mainly treats abdominal distension, and can stop diarrhea." "Kaibao Materia Medica" records that it "treats cold and heat and body swelling." "Compendium of Materia Medica" says: "Honeysuckle... people in the past called it the key medicine for treating wind and distension, relieving dysentery and expelling corpses... Later generations called it the key medicine for reducing swelling, dispersing toxins and treating sores." "Explanation of Medicinal Properties" records that it: "Honeysuckle... treats heat toxins and bloody dysentery, eliminates carbuncles and toxins, replenishes deficiency and treats wind." "Bencao Guizhen" says that it: "clears lung heat and resolves carbuncle toxins." It can be seen that honeysuckle clears away heat and dissipates wind, detoxifies and relieves sore throat, thereby achieving the effect of relieving throat discomfort.

[0049] Scrophularia: It tastes sweet, bitter and salty, and is slightly cold in nature. It enters the lung, stomach and kidney meridians and has the effects of clearing heat and cooling blood, purging fire and detoxifying, and nourishing yin. The Compendium of Materia Medica says: "It nourishes yin and reduces fire, relieves spots and toxins, benefits the throat, and relieves urinary blood stasis." The Bencao Zhengyi says: "Scrophularia, with the nature of extreme yin, is specialized in treating fever. The bitter taste can purge and descend, so it can treat symptoms such as heat accumulation in the internal organs. It tastes spicy and slightly salty, so it goes straight to the blood and relieves blood stasis. It can also flow externally to the meridians and marrow, and eliminate carbuncles and swelling caused by heat accumulation. It is cold but not harsh, moist but not greasy... It can dispel evil heat, but it is not a nourishing product after all." The Yuzhou Yaojie says: "Scrophularia, clears gold and replenishes water, and is good for all symptoms of sores, heat and pain, dryness and thirst in the chest and diaphragm, red and sticky urine, and bladder retention." "Xinglun" records that it: "can treat sudden heat, hot wind headache, typhoid fever, fatigue, and disperse tumors, goiters, and scrofula." "Bencao Gangmu" records that it: "treats wind-heat, toxic wind in the head, replenishes deficiency and fatigue, palpitations and irritability, fatigue and bone steaming, corpse evil spirits, stops forgetfulness, and eliminates swelling and poison." "Medical Origin" says that it: "treats heart annoyance and insomnia, confusion and despair, blood stasis and urination." "Pinhui Jingyao" says that it: "eliminates swelling of the throat and drains rootless fire." It can be seen that Scrophularia ningpoensis is an important medicine for nourishing yin, clearing heat, cooling blood and detoxifying. It can be used together with honeysuckle to enhance the effects of clearing heat, detoxifying and relieving sore throat.

[0050] Green fruit: It is mild in nature, sweet, sour and astringent in taste, and enters the lung and stomach meridians. It has the effects of clearing away heat and detoxifying, and relieving sore throat and promoting salivation. "Compendium of Materia Medica" says that it: "produces body fluid, relieves thirst, and treats sore throat. Chewing and swallowing the juice can detoxify all fish and turtle poisons." "Ben Jing Feng Yuan" records that it: "produces body fluid and quenches thirst, stimulates appetite and eliminates phlegm. It is suitable for coughs after being drunk or full, and for those with cold phlegm. People with smallpox should eat more, because it can detoxify and help the mild air in the stomach, causing the smallpox to break out." "Bencao Zaixin" says: "It calms the liver and stimulates appetite, moistens the lungs and nourishes yin, eliminates phlegm and regulates qi, stops coughs, and treats vomiting blood." "Lingnan Collection of Medicinal Materials" records: "The fruit can be boiled in water and washed to detoxify malnutrition and carbuncle." "Southern Yunnan Materia Medica" says: "It can treat all kinds of throat inflammation and big head plague. It can relieve damp heat, spring fever, produce body fluid and quench thirst, eliminate phlegm, and detoxify fish poison, alcohol, and stagnation." The green fruit can detoxify various poisons and has the effect of relieving sore throat and producing body fluid. It can be used together with honeysuckle and Scrophularia to enhance the detoxification and sore throat effects.

[0051] Platycodon grandiflorum: Pungent and bitter in flavor, neutral in nature, enters the Lung Meridian and has the effects of invigorating and promoting lung qi, dispelling phlegm and pus. "Ming Yi Bie Lu" states: "It benefits the five internal organs, intestines, and stomach, replenishes blood and qi, eliminates cold and heat, rheumatism, warms the middle and digests food, and treats sore throat and pharynx." "Zhenzhu Nang" states: "It treats sore throat, promotes lung qi, and treats nasal congestion." Platycodon grandiflorum invigorates and promotes lung qi, guiding other herbs upward to the pharynx. The formula primarily clears heat and fire, detoxifies and relieves sore throat, and is supplemented with herbs that nourish yin and cool blood, promote fluid production and relieve cough, and strengthen the spleen and transport fluid. This formula, combining both food and medicine, is formulated to aid in the treatment of chronic pharyngitis and alleviate various symptoms.

[0052] Phyllanthus emblica: Sweet, sour, astringent, and cooling, it enters the lung and stomach meridians. It has the effects of clearing heat and cooling blood, promoting digestion and strengthening the stomach, and promoting fluid production and relieving coughs. The "Compendium of Materia Medica" states that it "tonic and strengthens the body." The "Haiyao Bencao" states that it "tastes bitter, sour, sweet, and slightly cold, and treats lung damage caused by calamities and stones, shortness of breath and coughs. Long-term use can help reduce weight and prolong life." The "Compendium of Materia Medica" states that it "detoxifies metal and stone poisons and can be taken as a decoction." The "Compendium of Materia Medica" states that it "quenches thirst and promotes fluid production." The "Nanning City Pharmacopoeia" states that it "cools and detoxifies, and treats sore throat." Phyllanthus emblica, combined with Scrophularia ningpoensis and Cibotium barometz, enhances its heat-clearing and detoxifying effects, promotes lung qi, promotes fluid production and moistens the lungs, and relieves coughs and sore throats. The combined use of these herbs clears heat, detoxifies, relieves sore throats, promotes fluid production and moistens the lungs, and relieves coughs, effectively alleviating throat discomfort.

[0053] Luo Han Guo: Cool in nature, sweet in taste, enters the lung and large intestine meridians, and has the effects of clearing heat and moistening the lungs, relieving coughs and sore throats, and lubricating the intestines. "Lingnan Herbal Collection Records" records: "It treats coughs caused by phlegm and fire, and can be boiled with pork and taken as a decoction." "Guangxi Chinese Materia Medica" states: "It relieves coughs and clears heat, cools blood and moistens the intestines. It also treats coughs, dry blood, stomach heat, and constipation."

[0054] Menthol: aromatic, flavoring and carminative, mainly used to produce a cooling sensation on the skin or mucous membranes to relieve discomfort, especially pain. "Compendium of Materia Medica" records: "Mint is pungent and dispersing, cool and clearing, and is specialized in eliminating wind and dissipating heat. Therefore, it is an important medicine for headaches, head wind, eye, throat, mouth and teeth diseases, children's convulsions, scrofula, and scabies." "Pharmaceutical Chemistry" says: "Mint is pungent and dispersing, cool and clear in nature, and can clear the six yang meridians and eliminate wind evils caused by heat. It is sharp and light in nature, and is good at moving on the head and face. It is used to treat aphonia, mouth and teeth, and clear the throat." "Materia Medica Treatise" records that it: "Removes anger, induces toxic sweat, breaks blood and stops dysentery, and clears the joints." "Southern Yunnan Materia Medica" records that it: "Treats all typhoid headaches, cholera vomiting and diarrhea, carbuncle, ulcer, scabies, epilepsy and other sores." It can be seen that mint clears the head and eyes, eliminates swelling and relieves pain, and is used together with Momordica grosvenori to clear the throat, assisting other medicines in clearing heat and detoxifying, relieving sore throat and pain, and promoting fluid production and relieving cough.

[0055] The edible and medicinal Chinese medicine preparation of the present invention is based on the traditional Chinese medicine's understanding of the etiology and pathogenesis of chronic pharyngitis, formulates the treatment principles of clearing heat and relieving sore throat, nourishing yin and cooling blood, moistening the lungs and relieving cough, selects edible and medicinal Chinese medicines that are of the same origin as medicine and can clear heat and detoxify, nourish yin and reduce fire, promote lung function and relieve sore throat and cough, and is intended to assist in the treatment of chronic pharyngitis by clearing heat, promoting the production of body fluids, and relieving sore throat, thereby alleviating the symptoms of pharyngeal discomfort.

[0056] At the same time, the present invention uses total saponins and chlorogenic acid as the hallmark component indicators of the traditional Chinese medicine preparation, and adopts specific ultraviolet-visible spectrophotometry and high-performance liquid chromatography with good stability, good repeatability, high accuracy, and high precision to determine the content, and performs quality control on the traditional Chinese medicine preparation product, thereby ensuring the quality stability of the traditional Chinese medicine preparation and the effectiveness of the throat-clearing and throat-moistening function. The traditional Chinese medicine preparation with the throat-clearing and throat-moistening function provided by the present invention is suitable for people susceptible to chronic pharyngitis, such as teachers, tour guides, and specific personnel working in dusty environments, and can be taken for a long time. Its good effect replaces the side effects that may be caused by taking drugs, and a satisfactory effect is achieved.

[0057] To confirm the stability of the quality of the traditional Chinese medicine preparation of the present invention, the tablets prepared according to the method of Example 1 (hereinafter referred to as the composition of the present invention) were subjected to hygiene, effective ingredient (total saponins, chlorogenic acid) and stability tests (batches 20191201, 20191202, and 20191203). The results are shown in Tables 1 to 4 below.

[0058] Table 1 Test results at month 0

[0059]

[0060] Table 2 Test results in the first month

[0061]

[0062] Table 3 Test results in the second month

[0063]

[0064] Table 4 Test results at the third month

[0065]

[0066] In order to verify the scientificity, applicability and reproducibility of the detection method of the characteristic components (effective components) of the traditional Chinese medicine preparation of the present invention, the present invention conducted a methodological investigation on the ultraviolet spectrophotometry method for total saponins and the high performance liquid chromatography method for chlorogenic acid.

[0067] Investigation on the applicability of the determination method to this product

[0068] The inspection was conducted in reference to the Guiding Principles for Validation of Analytical Methods for Drug Quality Standards (General Rules 9101 of Part IV of the Pharmacopoeia of the People's Republic of China 2015 edition).

[0069] 1. Instrument:

[0070] 1.1 UV-visible spectrophotometer: Agilent Technologies Cary 60 UV-Vis;

[0071] 1.2 Ultrasonic cleaner: KQ-250B model from Kunshan Ultrasonic Instrument Co., Ltd.

[0072] 1.3 Analytical balance: METTLER TOLEDO XS204;

[0073] 1.4 Glass chromatography column: inner diameter 1.5 cm (same as the inner diameter of a 10 mL syringe)

[0074] 2 Reagents and materials

[0075] 2.1Amberlite-XAD-2 macroporous resin, Sigma Chemical Company, USA.

[0076] 2.2 Analytical grade ethanol

[0077] 2.3 For neutral alumina chromatography, 100-200 mesh.

[0078] 2.4 Vanillin solution Weigh 5 g of vanillin, dissolve it in glacial acetic acid and dilute to 100 mL.

[0079] 2.5 Perchloric acid analytical grade

[0080] 2.6 Glacial acetic acid analytical grade

[0081] 2.7 Ginsenoside Re was purchased from the China Food and Drug Administration, batch number: 110754-201626, for content determination, with a content of 97.4%. Tablets of the present invention (prepared according to Example 1): batch numbers 170201, 170202, and 170203.

[0082] 3. Preparation of Ginsenoside Re Standard Solution

[0083] Accurately weigh 20.45 mg of ginsenoside Re standard, dissolve it in methanol and make up to the volume in a 10 mL volumetric flask, that is, each mL contains 1.9918 mg of ginsenoside Re.

[0084] 4 wavelength scanning

[0085] The ginsenoside Re standard solution after color development was taken for spectral scanning at a wavelength of 400-800nm. The spectrum showed that there was strong absorption at 540-560nm, which is close to the detection wavelength of 560nm in the "Determination of total saponins in health foods" in the "Technical Specifications for Inspection and Evaluation of Health Foods" (2003 edition). Therefore, this product was determined at 560nm, and the method is feasible.

[0086] 5. Sample processing

[0087] 5.1 Investigation of sampling volume

[0088] Take an appropriate amount of this product, grind it into powder, mix it evenly, take 0.5, 1.0, 1.5, 2.0g, accurately weigh it, put it into a 100mL volumetric flask, add 70mL of water, ultrasonically treat it for 30 minutes, then dilute it to 100mL with water, shake it well, let it stand, and draw 1.0mL of the supernatant for column chromatography. At the same time, draw 50μL of ginsenoside Re standard solution (1.9918mg / mL) into an evaporating dish, evaporate it in a water bath (below 60℃), and dissolve it with a small amount of water. Column chromatography was then performed, and the sample and standard eluate were collected in an evaporating dish and evaporated in a 60°C water bath. 0.2 mL of 5% vanillin glacial acetic acid solution and 0.8 mL of perchloric acid were added, mixed, and transferred to a 5 mL stoppered graduated centrifuge tube. The tube was heated in a 60°C water bath for 10 minutes, removed, cooled in an ice bath, and 5.0 mL of glacial acetic acid was accurately added. After shaking, colorimetry was performed at a wavelength of 560 nm using a 1 cm colorimetric cell. The absorbance value was determined and the total saponin content was calculated. The results are shown in Table 5.

[0089] Table 5 Results of sample weighing

[0090]

[0091]

[0092] From the above results, it can be seen that the sample weight has little effect on the total saponins, but the sample absorbance values ​​are lower than 0.3 when the sample weight is 0.5g and 1.0g, which affects the accuracy of the measurement results. When the sample weight is 1.5g, the sample absorbance value exceeds 0.3. Therefore, in order to ensure the accuracy of the measurement, the sample weight is selected as 1.5g.

[0093] 5.2 Ultrasonic time investigation

[0094] Take an appropriate amount of this product, grind it into powder, mix it evenly, weigh 1.5g, a total of 4 parts, accurately weigh it, put it in a 100mL volumetric flask, add 70mL of water, ultrasonically treat it for 20, 30, 40, and 50 minutes respectively, then dilute it to 100mL with water, shake it well, let it stand, and draw 1.0mL of the supernatant for column chromatography respectively. At the same time, draw 50μL of ginsenoside Re standard solution (1.9918mg / mL) into an evaporating dish, evaporate it in a water bath (below 60℃), and add a small amount of water. After dissolution, column chromatography was performed, and the sample and standard eluate were collected in an evaporating dish and evaporated in a 60°C water bath. 0.2 mL of 5% vanillin glacial acetic acid solution and 0.8 mL of perchloric acid were added, mixed, and transferred to a 5 mL stoppered graduated centrifuge tube. The tube was heated in a 60°C water bath for 10 minutes, removed, cooled in an ice bath, and 5.0 mL of glacial acetic acid was accurately added. After shaking, colorimetry was performed at a wavelength of 560 nm using a 1 cm colorimetric cell. The absorbance was measured and the total saponin content was calculated. The results are shown in Table 6.

[0095] Table 6 Ultrasound time investigation results

[0096]

[0097] From the above results, it can be seen that the content of total saponins no longer increases after 40 minutes of ultrasound, indicating that the total saponins are completely extracted. Therefore, the ultrasound time is determined to be 40 minutes.

[0098] 6 Linear relationship investigation

[0099] Accurately pipette 30, 40, 50, 60, 80, and 100 μL of ginsenoside Re standard solution (1.9918 mg / mL) into an evaporating dish, evaporate in a water bath (below 60°C), dissolve with a small amount of water, and perform column chromatography. Collect the sample and standard eluate into an evaporating dish, evaporate in a 60°C water bath, add 0.2 mL of 5% vanillin glacial acetic acid solution and 0.8 mL of perchloric acid, mix well, and transfer into a 5 mL stoppered graduated centrifuge tube, heat in a 60°C water bath for 10 minutes, take out, cool in an ice bath, accurately add 5.0 mL of glacial acetic acid, shake well, and perform colorimetry at a wavelength of 560 nm using a 1 cm colorimetric cell to measure the absorbance. The results are shown in Table 7.

[0100] Table 7 Standard curve determination results

[0101]

[0102] The standard curve was drawn with the absorbance value as the vertical axis and the ginsenoside Re amount as the horizontal axis, and the regression equation was calculated: Y = 0.0045X + 0.0040, r = 0.9987. The results showed that the ginsenoside Re amount had a good linear relationship in the range of 59.754-199.180 μg.

[0103] 7. Precision inspection

[0104] Take an appropriate amount of this product, grind it finely, mix it evenly, weigh 1.5 g, accurately weigh it, place it in a 100 mL volumetric flask, add 70 mL of water, ultrasonicate it for 40 minutes, then dilute it to 100 mL with water, shake it well, let it stand, and draw 1.0 mL of the supernatant for column chromatography. At the same time, draw 50 μL of ginsenoside Re standard solution (1.9918 mg / mL) into an evaporating dish, evaporate it in a water bath (below 60°C), dissolve it with a small amount of water and then perform column chromatography. Collect the sample and standard eluate in an evaporating dish, evaporate it in a water bath at 60°C, add 0.2 mL of 5% vanillin glacial acetic acid solution and 0.8 mL of perchloric acid, mix well, transfer it to a 5 mL stoppered graduated centrifuge tube, heat it in a 60°C water bath for 10 minutes, take it out, cool it in an ice bath, accurately add 5.0 mL of glacial acetic acid, shake it well, and perform colorimetry at a wavelength of 560 nm using a 1 cm colorimetric cell. Measure the absorbance value 5 times in a row. The results are shown in Table 8.

[0105] Table 8 Precision test results

[0106]

[0107] From the above results, it can be seen that the absorbance values ​​of the standard and sample are stable, the RSD% value of the standard is 0.09%, and the RSD% value of the sample is 0.23%, indicating that the instrument has good precision.

[0108] 8. Stability investigation

[0109] The color-developed ginsenoside Re standard solution and sample solution were taken, and the absorbance values ​​were measured at a wavelength of 560 nm at regular intervals using a 1 cm colorimetric cell. The test results are shown in Table 9.

[0110] Table 9 Stability test results

[0111]

[0112] From the above results, it can be seen that the absorbance values ​​of the standard solution and sample solution decrease slightly with time. Therefore, the measurement should be carried out as soon as possible after color development to ensure the accuracy of the measurement results.

[0113] 9 Repeatability study

[0114] Take an appropriate amount of this product, grind it into powder, mix it evenly, weigh 1.5g, a total of 6 parts, accurately weigh it, put it in a 100mL volumetric flask, add 70mL of water, ultrasonically treat it for 40 minutes, then dilute it to 100mL with water, shake it well, let it stand, and draw 1.0mL of the supernatant for column chromatography. At the same time, draw 50μL of ginsenoside Re standard solution (1.9918mg / mL) and put it in an evaporating dish, evaporate it in a water bath (below 60℃), dissolve it with a small amount of water and then column chromatography. After chromatography, the sample and standard eluate were collected in an evaporating dish, evaporated to dryness in a 60°C water bath, 0.2 mL of 5% vanillin glacial acetic acid solution and 0.8 mL of perchloric acid were added, mixed, and transferred to a 5 mL stoppered graduated centrifuge tube, heated in a 60°C water bath for 10 minutes, removed, cooled in an ice bath, and 5.0 mL of glacial acetic acid was accurately added. After shaking, colorimetry was performed at a wavelength of 560 nm using a 1 cm colorimetric cell, the absorbance was determined, and the total saponin content was calculated. The results are shown in Table 10.

[0115] Table 10 Repeatability test results

[0116]

[0117] From the determination results, it can be seen that the RSD of total saponins is 2.21%, indicating that the method has good repeatability and is suitable for this product.

[0118] 10 Accuracy inspection

[0119] The sample recovery test was adopted. The same batch of samples were taken, ground and mixed evenly. Six portions were weighed, each of which was 0.75 g. The portions were accurately weighed and placed in a 100 mL volumetric flask. 25 mL of ginsenoside Re aqueous solution with a concentration of 0.114 μg / mL was accurately added. 45 mL of water was added and shaken. Ultrasonic treatment was performed for 40 minutes. The volume was then made up to 100 mL with water. The mixture was shaken and placed. 1.0 mL of the supernatant was respectively drawn for column chromatography. At the same time, 50 μL of ginsenoside Re standard solution (1.9918 mg / mL) was accurately drawn and placed in an evaporating dish. , evaporate in a water bath (below 60°C), dissolve in a small amount of water and perform column chromatography, collect the sample and standard eluate in an evaporating dish, evaporate in a water bath at 60°C, add 0.2mL of 5% vanillin glacial acetic acid solution and 0.8mL of perchloric acid, mix well and transfer into a 5mL stoppered graduated centrifuge tube, heat in a 60°C water bath for 10 minutes, take out, cool in an ice bath, accurately add 5.0mL of glacial acetic acid, shake well, perform colorimetry at a wavelength of 560nm with a 1cm colorimetric cell, determine the absorbance value, calculate the total saponin content, calculate the recovery rate, and the results are shown in Table 11.

[0120] Table 11 Recovery determination results

[0121]

[0122] From the determination results, it can be seen that the average recovery rate of total saponins is 96.65% (n=6) and the RSD is 2.96%. The determination method is accurate and the results are close to the true value. The method is suitable for this product.

[0123] 11 Investigation of detection limit

[0124] Accurately pipette 5, 10, 15, and 50 μL of ginsenoside Re standard solution (1.9918 mg / mL) into an evaporating dish, evaporate to dryness in a water bath (below 60°C), dissolve with a small amount of water, and perform column chromatography. Collect the eluate in an evaporating dish, evaporate to dryness in a 60°C water bath, add 0.2 mL of 5% vanillin glacial acetic acid solution and 0.8 mL of perchloric acid, mix well, and transfer to a 5 mL stoppered graduated centrifuge tube, heat in a 60°C water bath for 10 minutes, take out, cool in an ice bath, accurately add 5.0 mL of glacial acetic acid, shake well, and perform colorimetry at a wavelength of 560 nm using a 1 cm colorimetric cell to measure the absorbance. The results are shown in Table 12.

[0125] Table 12 Detection limit determination results

[0126]

[0127]

[0128] It can be seen from the measurement results that when the amount of ginsenoside Re is 19.918 μg, the measurement result deviates slightly from the actual content and can be reliably detected. When the amount of ginsenoside Re is 9.959 μg, the absorbance value is only 0.0535, and the measurement result deviates greatly from the actual content and cannot be reliably detected.

[0129] Therefore, based on the above test results, the detection limit of total saponins in this product was determined as follows: when the sample size was 1.5 g and the fixed volume was 100 mL, the detection limit of the method was 0.14 g / 100 g.

[0130] 12 Sample measurement

[0131] Take appropriate amount of three batches of samples, grind them respectively, mix them evenly, take 1.5g, take 2 parts of each batch, accurately weigh them, put them in 100mL volumetric flasks, add 70mL of water to each, ultrasonically treat for 40 minutes, then dilute to 100mL with water, shake well, let it stand, and draw 1.0mL of supernatant for column chromatography. At the same time, accurately draw 50μL of ginsenoside Re standard solution (1.9918mg / mL) and put it in an evaporating dish, evaporate it in a water bath (below 60℃), and use a small amount of water to dissolve it. After decomposition, column chromatography was performed, and the sample and standard eluate were collected in an evaporating dish and evaporated in a 60°C water bath. 0.2 mL of 5% vanillin glacial acetic acid solution and 0.8 mL of perchloric acid were added, mixed, and transferred to a 5 mL stoppered graduated centrifuge tube, heated in a 60°C water bath for 10 minutes, taken out, cooled in an ice bath, and 5.0 mL of glacial acetic acid was accurately added. After shaking, colorimetry was performed at a wavelength of 560 nm using a 1 cm colorimetric cell, the absorbance value was determined, and the total saponin content was calculated. The results are shown in Table 13.

[0132] Table 13 Measurement results of three batches of samples

[0133]

[0134] The average contents of the three batches of samples were 0.4516g / 100g, 0.4310g / 100g and 0.4558g / 100g respectively, and the method is feasible.

[0135] Conclusion: The method for determining the total saponins content of this product was validated by referring to the guidelines for validation of analytical methods for pharmaceutical quality standards (9101, Part IV, General Rules of the Pharmacopoeia of the People's Republic of China, 2015 edition) and carrying out methodological validation tests such as sample sampling amount, linear relationship investigation, precision investigation, stability investigation, repeatability investigation, and accuracy investigation. The results showed that the detection method adopted was suitable for the determination of total saponins of this product.

[0136] Methodological investigation on the determination of chlorogenic acid by high performance liquid chromatography

[0137] Investigation on the applicability of the determination method to this product

[0138] The inspection was conducted in reference to the Guiding Principles for Validation of Analytical Methods for Drug Quality Standards (General Rules 9101 of Part IV of the Pharmacopoeia of the People's Republic of China 2015 edition).

[0139] 1 Instrument

[0140] High performance liquid chromatograph (with UV detector): Agilent 1100.

[0141] Ultrasonic cleaner: KQ-250B produced by Kunshan Ultrasonic Instrument Co., Ltd.

[0142] Analytical balance: METTLER TOLEDO XS204.

[0143] 2 Reagents and materials

[0144] Column: Waters Symmetry C 18 5μm 4.6×250mm.

[0145] The mobile phase used acetonitrile of chromatographic grade, other reagents were of analytical grade, and water was ultrapure water.

[0146] Tablets of the present invention: batch numbers 170201 170202 170203.

[0147] Chlorogenic acid standard: purchased from China Food and Drug Inspection Institute, batch number: 110753-201314 for content determination, the content is calculated as 96.6%.

[0148] 3. Preparation of standard solution

[0149] Take an appropriate amount of chlorogenic acid standard, add mobile phase to make a solution containing 80.6610 μg of chlorogenic acid per 1 mL, shake well, and obtain the solution.

[0150] 4. Detection wavelength investigation

[0151] The standard solution was taken and spectral scanning was performed in the wavelength range of 200-800nm. The results showed that the maximum absorption at a wavelength of 329nm was not much different from the wavelength included in the national standard, and the detection wavelength was determined to be 327nm.

[0152] 5 Chromatographic conditions

[0153] Mobile phase: 0.5% acetic acid solution + acetonitrile = 9 + 1

[0154] Flow rate: 1.0 mL / min

[0155] Column temperature: 35°C

[0156] Detection wavelength: 327nm

[0157] Injection volume: 10 μL

[0158] 6 Linear relationship investigation

[0159] Accurately pipette 10 μl of chlorogenic acid standard solution with concentrations of 2.0165 μg / mL, 10.0825 μg / mL, 20.1653 μg / mL, 40.3305 μg / mL, and 80.6610 μg / mL, respectively, inject into the liquid chromatograph, and measure the peak area according to the specified chromatographic conditions. The results are shown in Table 14.

[0160] Table 14 Standard curve determination results

[0161]

[0162]

[0163] The standard curve was drawn with the peak area integral value as the ordinate and the injection volume as the abscissa: the regression equation was calculated as: Y = 2.803908X-14.076693, r = 0.9999. The results showed that the injection volume of the chlorogenic acid standard had a good linear relationship in the range of 20.165ng-806.610ng.

[0164] 7. Specificity inspection

[0165] (1) Preparation of negative samples

[0166] Take the other medicinal ingredients in the prescription except honeysuckle, prepare negative samples according to the preparation process, treat the negative samples according to the sample processing method, prepare negative sample solutions, and perform peak detection.

[0167] (2) Test results

[0168] Accurately pipette 10 μl of each of the standard solution, sample solution, and negative sample solution, and perform the determination under the selected chromatographic conditions. The results show that the chromatographic peaks have good separation, and the negative sample has no interference, indicating that the other medicinal flavors in the prescription, except honeysuckle, do not interfere with the determination of chlorogenic acid. The results are as follows: Figure 1 shown.

[0169] 8. Sample processing

[0170] (1) Investigation of sample weight

[0171] The tablets of the present invention were taken, ground into powder, and mixed evenly. 0.1, 0.2, 0.3, 0.4, and 0.5 g were respectively weighed, accurately weighed, and placed in a 25 mL volumetric flask. 20 mL of 70% methanol was added and ultrasonicated for 30 minutes. The volume was adjusted to the scale with 70% methanol, shaken, and filtered through a 0.45 μm microporous membrane. The filtrate was subjected to liquid chromatography analysis, and the chlorogenic acid content was determined according to the method. The results are shown in Table 15.

[0172] Table 15 Results of sample weighing

[0173]

[0174] From the above results, it can be seen that the sample weight has little effect on chlorogenic acid. When the sample weight is 0.2 g, the peak area of ​​the sample is moderate and basically in the middle of the standard curve. Therefore, the sample weight is selected as 0.2 g.

[0175] (2) Investigation of ultrasound time

[0176] The tablets of the present invention were ground into powder and mixed evenly. 0.2 g of each was weighed and accurately weighed into four portions. The mixture was placed in 25.0 mL volumetric flasks, and 20 mL of 70% methanol was added to each portion. The mixture was ultrasonically treated for 10, 20, 30, and 40 minutes, respectively. The mixture was taken out, cooled, and the volume was adjusted to the mark with 70% methanol. The mixture was shaken and filtered through a 0.45 μm filter membrane. The filtrate was analyzed by liquid chromatography. The content of chlorogenic acid was determined according to the method. The results are shown in Table 16.

[0177] Table 16 Ultrasonic time investigation results

[0178]

[0179] From the test results, it can be seen that the content difference at different ultrasonic times is not much, but the content is highest when ultrasonicated for 30 minutes. In order to ensure the complete extraction of chlorogenic acid, the ultrasonic extraction time is selected as 30 minutes.

[0180] 9. Precision inspection

[0181] 10 μL of chlorogenic acid standard solution and sample solution were taken and injected 5 times continuously. The peak area was measured according to the determined chromatographic conditions. The test results are shown in Table 17.

[0182] Table 17 Precision test results

[0183]

[0184] From the above results, it can be seen that the peak areas of the standard and sample are stable, the RSD% value of the standard is 0.42%, and the RSD% value of the sample is 0.36%, indicating that the instrument has good precision.

[0185] 10. Stability Investigation

[0186] The standard solution and the corresponding sample solution were aspirated and injected once at regular intervals to determine the peak area. The test results are shown in Table 18.

[0187] Table 18 Stability test results

[0188]

[0189]

[0190] From the above results, it can be seen that within 24 hours of detection, the peak areas of the standard and sample were stable, with RSD values ​​of 1.65% and 1.69%, respectively.

[0191] 11 Repeatability Study

[0192] Take the same batch of samples, grind them into powder, mix them evenly, take three sample sizes of high, medium and low respectively, 3 portions of each sample size, prepare them separately according to the sample preparation method, and measure them according to the method. The measurement results are shown in Table 19.

[0193] Table 19 Repeatability test results

[0194]

[0195] From the determination results, it can be seen that the RSD of chlorogenic acid content is 1.27%, indicating that the method has good repeatability and is suitable for this product.

[0196] 12 Accuracy inspection

[0197] A sample recovery experiment was conducted using nine samples from the same batch, divided into three groups. 20 ml of a high, medium, and low concentration chlorogenic acid reference solution was precisely added to each group. Ultrasonic extraction was performed, and the sample was measured and the recovery rate was calculated. The results are shown in Table 20.

[0198] Table 20 Recovery determination results

[0199]

[0200]

[0201] From the determination results, it can be seen that the average recovery rate of chlorogenic acid is 97.68% (n=9) and the RSD is 1.85%. The determination method is accurate and the results are close to the true value. The method is suitable for this product.

[0202] 13 Durability inspection

[0203] 13.1 Effect of Detection Wavelength Change on Content

[0204] Take the same sample and change the detection wavelength for measurement. The test results are shown in Table 21.

[0205] Table 21 Content determination results of the same sample with different detection wavelengths

[0206]

[0207] From the above results, it can be seen that the RSD of the content of the same sample measured by changing the detection wavelength is 0.17%.

[0208] 13.2 Effect of Column Temperature Change on Assay

[0209] Take the same sample and change the column temperature for measurement. The test results are shown in Table 22.

[0210] Table 22 Column temperature changes for the same sample content determination results

[0211]

[0212] From the above results, it can be seen that the RSD of the content of the same sample measured with column temperature change is 0.42%.

[0213] 13.3 Effect of Changing the Mobile Phase Ratio on Assay

[0214] Take the same sample and change the ratio of mobile phase for determination. The test results are shown in Table 23.

[0215] Table 23 Content determination results of the same sample with changes in mobile phase ratio

[0216]

[0217] From the above results, it can be seen that when the mobile phase ratio changes slightly, the RSD of the content change of the same sample is 0.32%.

[0218] 13.4 Effect of Different Chromatographic Columns on Content

[0219] The same sample was taken and chromatographic columns from different manufacturers were used for determination. The test results are shown in Table 24.

[0220] Table 24 Content determination results of the same sample using different chromatographic columns

[0221]

[0222] From the above results, it can be seen that when different chromatographic columns are used to measure the same sample, the RSD of the content is 2.74%.

[0223] The results of the durability test showed that the detection results of chlorogenic acid content were slightly affected by different flow rates, different mobile phase ratios, different column temperatures and different chromatographic columns, indicating that the method is feasible.

[0224] 14 Detection limit investigation

[0225] A chlorogenic acid standard solution (C = 85.9740 μg / mL) was taken and diluted with the mobile phase to prepare solutions with concentrations of 42.9870 μg / mL, 21.4935 μg / mL, 10.7468 μg / mL, 2.1494 μg / mL, 1.0747 μg / mL, and 0.5374 μg / mL, respectively. The detection limit was investigated. 10 μl of the chlorogenic acid standard solution of each concentration was accurately aspirated and injected into the liquid chromatograph. The peak area was determined according to the specified chromatographic conditions.

[0226] When the injection volume was 5.374 ng, the peak area was only 7.104, which deviated greatly from the actual value and could not be reliably detected.

[0227] When the injection volume was 10.747 ng, the peak area was 22.837, and regression calculation could be performed. The data was reliable, and the regression equation was Y=2.34602X-8.93993, r=0.99996.

[0228] Therefore, based on the above test results, the detection limit of chlorogenic acid in this product is determined as follows: when the sample size is 0.2 g and the fixed volume is 25 mL, the detection limit of the method is 0.014 g / 100 g.

[0229] 15 Sample measurement

[0230] Take 10 tablets of each of three batches of samples, grind them into powder, mix them evenly, take 0.2 g of each, weigh them accurately, place them in 25.0 mL volumetric flasks, add 20 mL of 70% methanol to each, treat them ultrasonically for 30 minutes, take them out, cool them, make up to the volume with 70% methanol, shake them well, pass them through a 0.45 μm filter membrane, and analyze the filtrate by liquid chromatography. The results are shown in Table 25.

[0231] Table 25 Three batches of sample content determination results

[0232]

[0233]

[0234] The average contents of the three batches of samples were 4.062 mg / g, 4.050 mg / g and 4.080 mg / g, respectively. The method is feasible.

[0235] Conclusion: The method for determining the chlorogenic acid content of this product was verified by referring to the Guiding Principles for Validation of Analytical Methods for Pharmaceutical Quality Standards (General Rules 9101 of Part IV of the Pharmacopoeia of the People's Republic of China 2015 edition), and methodological validation tests including sample spectrum scanning, sampling volume, ultrasonic time, linear relationship investigation, specificity investigation, precision investigation, stability investigation, repeatability investigation, accuracy investigation, and durability investigation were carried out. The results showed that the detection method adopted was suitable for the determination of chlorogenic acid in this product.

[0236] To confirm the effects of the Chinese medicinal preparation of the present invention in clearing the throat, moistening the throat, treating chronic pharyngitis, and relieving pharyngeal discomfort, the following functional tests were conducted using the active component prepared according to the method of Example 1 (hereinafter referred to as the composition of the present invention).

[0237] Throat clearing animal function test:

[0238] The Analytical and Evaluation Center of West China School of Public Health, Sichuan University was commissioned to conduct a throat clearing animal function test on the composition of the present invention. The test results showed that:

[0239] The mice (Balb / c) were orally administered with the composition of the present invention at doses of 0.8 g / kg.bw, 1.6 g / kg.bw and 2.4 g / kg.bw (equivalent to 10 times, 20 times and 30 times of the recommended daily dosage recommended by the submitting party respectively), and the SD rats were administered at doses of 0.4 g / kg.bw, 0.8 g / kg.bw and 1.6 g / kg.bw (equivalent to 5 times, 10 times and 20 times of the recommended daily dosage recommended by the submitting party respectively). After 30 days, the tests were carried out. In the mouse ear swelling test, compared with the control group of mice, the auricle swelling rate of the 2.4 g / kg.bw dose group was significantly reduced. In the rat cotton ball implantation test, compared with the control group, the net amount of granuloma in the 1.6 g / kg.bw dose group was significantly reduced; in the rat foot swelling test, compared with the control group, the swelling rate of the toe volume at 1 h in the 0.8 g / kg.bw dose group was significantly reduced. According to the evaluation criteria, it can be considered that the throat-clearing function test of this sample is a positive result.

[0240] 1 Materials and Methods

[0241] 1.1 The composition of the present invention was provided by Shijiazhuang Yiling Pharmaceutical Co., Ltd., and the recommended daily dosage recommended by the submitting party was 4.8 g / person / day.

[0242] 1.2 Experimental animals

[0243] SPF-grade male Ba1b / c mice, weighing from 18 g to 22 g, were provided by Chengdu Dashuo Biotechnology Co., Ltd. (license number: SCXK (Sichuan) 2013-24). They were raised in a barrier-level animal house (license number: SYXK (Sichuan) 2013-011 of the Sichuan Experimental Animal Management Committee). The drinking water was purified water. During the whole test process, the animals were allowed to freely feed and drink water. The room temperature was 20°C - 24°C, and the relative humidity was 45% - 65%.

[0244] SPF-grade male SD rats, weighing from 180 to 220 g, were provided by Chengdu Dashuo Biotechnology Co., Ltd. (license number: SCXK (Sichuan) 2013-24). They were raised in a barrier-level animal house (license number: SYXK (Sichuan) 2013-011 of the Sichuan Experimental Animal Management Committee). The drinking water was purified water. During the whole test process, the animals were allowed to freely feed and drink water. The room temperature was 20°C - 24°C, and the relative humidity was 45% - 65%.

[0245] 1.3 Dose selection

[0246] Mice were purchased and fed a basal diet before being randomly divided into 0.8g / kg.bw, 1.6g / kg.bw, and 2.4g / kg.bw groups (equivalent to 10, 20, and 30 times the recommended daily dose, respectively) and a control group, with 12 mice in each group. The experimental groups were gavaged with the corresponding dose of sample (12g of the sample was weighed, thoroughly ground with a small amount of sterile deionized water, and then mixed with sterile deionized water to 100mL. This served as the high-dose group gavage solution: 1 part sterile water and 2 parts high-dose group gavage solution were mixed to form the medium-dose group gavage solution; the medium-dose group gavage solution was diluted 1 / 2 with sterile water and mixed to form the low-dose group gavage solution). The control group was gavaged with sterile water, with a volume of 20ml / kg.bw, once daily for 30 consecutive days. Rats were purchased and fed a basal diet before being randomly divided into 0.4g / kg.bw, 0.8g / kg.bw, and 1.6g / kg.bw dose groups (equivalent to 5, 10, and 20 times the recommended daily dose, respectively) and a control group, with 24 rats in each group. The experimental groups received the corresponding dose of the sample (16g of the sample was weighed, thoroughly ground with a small amount of sterile deionized water, and then sterile deionized water was added to 100°C and mixed thoroughly to serve as the high-dose group gavage solution; 1 part sterile water was mixed with 2 parts of the high-dose group gavage solution to serve as the medium-dose group gavage solution; the medium-dose group gavage solution was diluted 1 / 2 times with sterile water and mixed thoroughly to serve as the low-dose group gavage solution). The control group received sterile water gavage at a volume of 10ml / kg.bw once daily for 30 consecutive days.

[0247] 1.4 Main instruments and reagents

[0248] 1.4.1 Main instruments: BSA224S electronic balance; Sartorius AG, Germany; toe volume measuring instrument; Chengdu Taimeng Technology Co., Ltd.; 9mm diameter hole punch, etc.

[0249] 1.4.2 Main reagents: Dextran 40,000: Shanghai Future Industrial Co., Ltd.; Xylene: Tianjin Ruijinte Chemical Co., Ltd.

[0250] 1.5 Experimental methods

[0251] 1.5.1 Body weight determination: The animals were gavage-fed for 30 consecutive days, and their body weights were measured on the 1st, 8th, 15th, 23rd, and 30th day after gavage.

[0252] 1.5.2 Mouse Ear Swelling Test: On the 30th day of continuous gavage, 20 μl of xylene was dripped onto the center of the auricle on the outer side of the right ear of the mouse and allowed to diffuse freely. After 30 minutes, the mouse was euthanized, and both ears were clipped. A 9 mm diameter punch was used to punch out the same part of each ear and weighed. The difference in weight between the two ears was used as the auricle swelling value, and the auricle swelling rate was calculated. If the auricle swelling rate in the experimental group was significantly reduced compared to the control group, the difference was statistically significant, indicating that the test sample had a positive result in the mouse ear swelling test.

[0253] Auricle swelling rate (%) = auricle swelling value / control ear piece weight x 100%

[0254] 1.5.3 Rat Cotton Ball Implantation Test: Cotton balls of approximately 20-25 mg were made into compact balls. After autoclaving, the balls were placed in a constant temperature drying oven at 60°C for 3 h, taken out and weighed under aseptic conditions, and dried for later use. Eight days before the end of the experiment, the hair on both sides of the rat's groin was removed with an epilator. The rats were lightly anesthetized with ether and disinfected with iodine. The skin on both sides of the rat's groin was incised under aseptic conditions, and spare cotton balls were implanted. The incisions were sutured, and the test substance was continued to be administered. On the day of the end of the experiment, the rats were euthanized 1 hour after the test substance was administered. The skin was cut open at the original suture site, and the cotton ball granulation tissue was peeled off and removed. The granuloma was placed in a weighed clean dish and dried in a constant temperature drying oven at 60°C with the lid open for 1 hour. The weight was weighed and the net amount of granuloma was calculated.

[0255] Compared with the blank control group, the net amount of granuloma in the experimental group was significantly reduced. The difference was statistically significant, which indicated that the cotton ball implantation test result of the test sample was positive.

[0256] Net amount of granuloma (mg) = weight of granuloma on dried cotton ball minus weight of original cotton ball

[0257] 1.5.4 Rat toe swelling test: The test sample was administered once by gavage on the day the experiment ended. One hour later, the volume of the right hind toes of each group of rats was measured with a toe volume meter, which was used as the toe volume at 0 hours. Then, 40,000 0.1 mL / rat of 1% dextran prepared with sterile steamed water was injected subcutaneously into the right hind toes of the rats. The volume of the rat toes was measured at 1, 2, 4, and 6 hours respectively. The same part was measured 3 times and the average value was taken. The difference between the toe volume measured at different times and the toe volume before the action of the inflammatory agent was taken as the swelling value, and the toe swelling rate of each time period was calculated. Compared with the control group, the toe volume swelling rate of the experimental group before and after stimulation at any time point was significantly reduced, and the difference was significant. It can be determined that the test result of the rat toe swelling test of the test sample was positive. Swelling rate (%) = swelling value / toe volume before inflammation x 100%

[0258] 1.6 Experimental data statistics

[0259] The data were processed by analysis of variance comparing the means of the experimental and control groups. The software used was the statistical software package of the "Chinese Medical Encyclopedia Medical Statistics" (PEMS3.1, Department of Health Statistics, West China Public Health, Sichuan University).

[0260] 2 Results

[0261] 2.1 Effect of the Composition of the Present Invention on Body Weight

[0262] 2.1.1 Mouse ear swelling group Variance analysis showed that there were no significant differences in initial body weight, mid-term body weight, final body weight, and body weight gain between each dose group and the control group (P>0.05), see Table 26.

[0263] Table 26 Effect of the composition of the present invention on body weight of mice with ear swelling

[0264]

[0265] 2.1.2 Cotton ball implantation group of rats According to variance analysis, there were no significant differences in initial body weight, mid-term body weight, final body weight, and body weight gain between each dose group and the control group (P>0.05), see Table 27.

[0266] Table 27 Effect of the composition of the present invention on body weight of rats in the cotton ball implantation group

[0267]

[0268] 2.1.3 Rat toe swelling group Variance analysis showed that there were no significant differences in initial body weight, mid-term body weight, final body weight, and body weight gain between each dose group and the control group (P>0.05), see Table 28.

[0269] Table 28 Effect of the composition of the present invention on body weight of rats with swollen toes

[0270]

[0271] 2.2 Effect of the composition of the present invention on the swelling rate of the mouse ear

[0272] After variance analysis, the ear swelling rate of the test substance 0.8g / kg.bw and 1.6g / kg.bw groups were not significantly reduced compared with the control group mice (P>0.05), and the ear swelling rate of the test substance 2.4g / kg.bw group was significantly reduced (P<0.01), see Table 29.

[0273] Table 29 Effect of the composition of the present invention on the swelling rate of the mouse ear

[0274]

[0275] ** indicates P < 0.01 compared with the control group.

[0276] 2.3 Effect of the composition of the present invention on the net amount of granuloma in rats

[0277] After variance analysis, there was no significant difference in the net volume of granulomas between the 0.4g / kg.bw and 0.8g / kg.bw g / kg.bw dose groups and the control group rats (P>0.05), and the net volume of granulomas in the 1.6g / kg.bw dose group was significantly reduced compared with the control group rats (P<0.05), see Table 30.

[0278] Table 30 Effect of the composition of the present invention on the net amount of granuloma in rats

[0279]

[0280] * indicates P < 0.05 compared with the control group.

[0281] 2.4 Effect of the composition of the present invention on the swelling rate of rat toes

[0282] After variance analysis, there was no significant difference in the 2h swelling rate, 4h swelling rate, and 6h swelling rate between each dose group and the control group (P>0.05); in the 1h swelling rate determination, the 0.8g / kg.bw dose group was significantly lower than the control group (P<0.05), and there was no significant difference between the other two dose groups and the control group (P>0.05), see Table 31.

[0283] Table 31 Effect of the composition of the present invention on rat toe swelling rate

[0284]

[0285] * indicates P < 0.05 compared with the control group.

[0286] 3 Summary

[0287] Oral administration of the composition of the present invention to mice (Balb / c) at doses of 0.8g / kg.bw, 1.6g / kg.bw, and 2.4g / kg.bw for 30 days showed no adverse effects on body weight and weight gain; administration of the composition of the present invention to SD rats at doses of 0.4g / kg.bw, 0.8g / kg.bw, and 1.6g / kg.bw for 30 days showed no adverse effects on body weight and weight gain. In the mouse ear swelling test, the auricle swelling rate of the 2.4g / kg.bw dose group was significantly reduced compared with the control group (P < 0.01). In the rat cotton ball implantation test, the net volume of granulomas in the 1.6g / kg.bw dose group was significantly reduced compared with the control group (P < 0.05); in the rat toe swelling test, the 1h toe volume swelling rate was significantly reduced in the 0.8g / kg.bw dose group compared with the control group (P < 0.05). According to the evaluation criteria, the test product was considered to have a positive result in the animal experimental throat clearing function test.

[0288] Human trials on throat clearing function:

[0289] To verify the throat-clearing function of the present invention's composition, a human throat-clearing test was conducted at the Affiliated Hospital of West China University of Medical Sciences. The results are as follows: 101 eligible subjects were randomly divided into a trial group and a control group; 100 effective cases were found, with 50 in the trial group and 50 in the control group. After the trial group took the present invention's composition as directed for 30 days (the control group was a blank control), the improvement rates of the main clinical symptoms and signs in the trial group were 18.00% and 84.00%, respectively, while the improvement rates in the control group were 0.06% and 38.00%, respectively. The Z-comparison between the two groups showed significant differences (P < 0.05). The main clinical symptom and sign scores in the trial group decreased after the trial, and the difference was statistically significant compared with the control group before and after the trial (P < 0.05). Blood, urine, stool, and biochemical parameters in both groups were within normal ranges before and after the trial. No allergic reactions or other adverse reactions were observed during the trial. This indicates that this product has no adverse effects on the subjects' health.

[0290] According to the criteria of the throat clearing function evaluation method (National Food and Drug Administration

[2012] No. 107), the composition of the present invention has the throat clearing function.

[0291] 1 Materials and Methods

[0292] 1.1 Sample

[0293] The sample is the composition of the present invention, provided by Shijiazhuang Yiling Pharmaceutical Co., Ltd. and manufactured by the same company. The sample is a brown to tan tablet, weighing 0.8g per tablet, 18 tablets per bag, and the sample quantity is 600 bags. Batch number: 140701. Storage: Store sealed in a dry place. Shelf life: 24 months. The recommended dosage for humans is 2 tablets three times daily, for a daily dosage of 4.8g.

[0294] 1.2 Subjects

[0295] 101 subjects were selected on a voluntary basis.

[0296] 1.2.1 Subject inclusion criteria

[0297] Voluntary subjects who meet at least one of the following requirements 1.2.1.1 and 1.2.1.1.2 upon examination.

[0298] 1.2.1.1 Signs: Chronic pharyngitis, subjective symptoms include sore throat, itchy throat, dry throat, dry cough, foreign body sensation, aggravated by talking too much, etc.

[0299] 1.2.1.2 Pharyngeal symptoms: edema of the pharyngeal mucosa, congestion of the mucosa, hyperplasia of lymphoid follicles on the posterior pharyngeal wall, and adhesion of secretions.

[0300] 1.2.2 Subject Exclusion Criteria

[0301] Patients with any of the following conditions cannot be included in this trial.

[0302] 1.2.2.1 Acute attack of chronic pharyngitis or caused by acute pharyngitis, vocal cord nodules, cold or smoking.

[0303] 1.2.2.2 Those caused by lesions of the nasopharynx, pharynx, larynx, nose, larynx, esophagus, neck, tuberculosis, and metastatic lung cancer.

[0304] 1.1.2.3 Individuals under 18 years old or over 65 years old, pregnant or lactating women, and those who are allergic to the test product.

[0305] 1.2.2.4 Patients with serious diseases of the heart, cerebrovascular system, liver, kidney, hematopoietic system, bronchus and lungs, as well as mental illnesses and sleep disorders.

[0306] 1.2.2.5 Those who have taken items related to the function being tested in the short term, which may affect their judgment of the results.

[0307] 1.2.2.6 Those who fail to take the test sample as prescribed or take other drugs during the test, and the efficacy cannot be determined or the data is incomplete.

[0308] 1.3 Experimental Design and Grouping

[0309] Both self-controlled and intergroup controlled designs were used. Participants were randomly divided into a trial group and a control group based on their pharyngeal symptoms and signs. When grouping, factors that may influence the results, such as disease duration, age, and gender, were considered to the greatest extent possible. Balance tests were performed to ensure comparability between groups. Each group had at least 50 valid subjects.

[0310] 1.4 Dosage and time of consumption

[0311] The trial group took the composition of the present invention orally as recommended, three times daily, two tablets each time. A blank control group was used, i.e., no special requirements were made for the control group. The test substance was taken for 30 consecutive days. During the trial period, the subjects did not change their original dietary habits, maintained a normal diet, or take any medications or other health foods that promote throat clearing.

[0312] 1.5 Requirements for main instruments, reagents and testing environment: Hitachi 7180 fully automatic biochemical analyzer (Hitachi High-Tech Co., Ltd., Japan); DIRUI H-300 eight-item urine analyzer (Changchun Bianrui Company); Sysmex XN-I0[B2] three-category hematology analyzer (Csysmex Company); Sysmex blood cell analysis diluent (Sysmex Company); KX-21 fully automatic hematology analyzer (Sysmex Company, Japan); BS-800 fully automatic biochemical analyzer (Shenzhen Mindray Company); DIRUI urine analysis reagent strip (Changchun Dirui Company); Leadman biochemical test kit (Beijing Leadman Biochemical Co., Ltd.).

[0313] 1.6 Observation indicators

[0314] 1.6.1 General Conditions

[0315] Including spirit, sleep, diet, urination and defecation, heart rate, blood pressure, etc.

[0316] 1.6.2 Safety Observation

[0317] 1.6.2.1 Routine examination of blood, urine and stool: red blood cell count, hemoglobin, white blood cell count, platelet count, urine routine, stool microscopy, etc.

[0318] 1.6.2.2 Liver and kidney function tests: serum albumin (ALB), total protein (TP), liver and kidney function (aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), creatine urea (CRE), blood glucose (GLU), blood lipids (total cholesterol (TC), triglycerides (TG).

[0319] 1.6.2.3 Chest X-ray, electrocardiogram, and abdominal ultrasound: Perform one examination before the food tasting.

[0320] 1.6.3 Efficacy Observation

[0321] 1.6.3.1 Clinical symptom observation

[0322] Accurately record the subjective pharyngeal symptoms of the subjects before and after the food trial. The main pharyngeal symptoms include: pharyngeal pain, pharyngeal itching, pharyngeal dryness, dry cough, foreign body sensation aggravated by talking, etc. Calculate the score according to the severity of the symptoms (1 degree - 1 point, 2 degrees - 2 points, 3 degrees - 3 points), and calculate the change in score and symptom improvement rate.

[0323] 1.6.3.2 Physical signs observation

[0324] All subjects underwent a pharyngeal examination before and after the trial. Pharyngeal examinations detected signs such as pharyngeal mucosal congestion, mucosal edema, hyperplasia of the posterior pharyngeal lymphoid follicles, and secretions. Results were classified as mild, moderate, or severe, and were categorized as I, II, or III. Changes in physical signs before and after the trial were recorded, and the physical sign score and improvement rate were calculated.

[0325] 1.6.3.3 Efficacy determination

[0326] Effective: Symptoms were alleviated by 1 degree, and pharyngeal signs were alleviated by 1 grade.

[0327] Ineffective: There was no significant change in symptoms and signs.

[0328] 1.7 Data Statistical Analysis

[0329] Quantitative data were analyzed using the t-test. Paired t-tests were used for self-comparison, and two-group means were compared using the two-sample t-test. For data with non-normal distribution or uneven variance, appropriate variable transformation was performed. Once normality and homogeneity of variance were achieved, the transformed data were used for the t-test. If the transformed data still did not meet the normality and homogeneity of variance requirements, the rank sum test was used instead.

[0330] The improvement rate is count data, using x 2 test.

[0331] 1.8 Result determination

[0332] Comparison between the trial group before and after the trial and between the trial group and the control group after the trial showed that the clinical symptom scores and physical sign scores of the pharynx were significantly reduced, and the improvement rate of symptoms and signs was significantly increased compared with the control group. The differences were statistically significant, and it can be determined that the test sample has the function of clearing the pharynx.

[0333] 2 Results

[0334] 2.1 General Condition

[0335] This trial enrolled 101 subjects, who were randomly divided into a trial group (51 subjects) and a control group (50 subjects). One subject dropped out of the trial group during the trial, resulting in a total of 100 subjects, including 50 in the trial group and 50 in the control group. Pre-trial blood, urine, stool, liver and kidney function, chest X-ray, electrocardiogram, and ultrasound examinations were all within normal ranges. There were no significant differences in age, gender, disease duration, or scores of major clinical symptoms between the two groups before the trial (P > 0.05). A comparison of general conditions before the trial is shown in Table 32.

[0336] Table 32 Comparison of general conditions before tasting

[0337]

[0338] 2.2 Impact on efficacy indicators

[0339] 2.2.1 Symptom score and clinical symptom improvement rate

[0340] Comparisons of clinical symptom scores before and after the trial and between groups are shown in Table 33. There was no significant difference in clinical symptom scores between the trial group and the control group before the trial (P>0.05). There was no statistically significant difference in clinical symptom scores between the control group before and after the trial (P>0.05). The scores of major clinical symptoms in the trial group were significantly reduced before and after the trial and after the trial compared with the control group (P<0.05). After the trial, the overall improvement rates of clinical symptoms in the trial group and the control group were 0.06% and 18.00%, respectively, with the improvement rate in the trial group being significantly higher than that in the control group (P<0.05). The results are shown in Table 34.

[0341] Table 33 Changes in the main clinical symptom scores (integral value, )

[0342]

[0343] * indicates P < 0.05 for comparison before and after itself; # indicates P < 0.05 for comparison with the control group.

[0344] Table 34 Improvement of clinical symptoms

[0345]

[0346] * indicates P < 0.05 compared with the control group.

[0347] 2.2.2 Physical sign scores and physical sign improvement rate

[0348] The results of the physical sign score analysis are shown in Table 35. After the trial diet, the physical sign scores of the trial group were significantly reduced, with significant differences compared to the control group before and after the trial diet (P < 0.05). The overall improvement rates of major physical signs in the control group and the trial diet group were 38.00% and 84.00%, respectively, with the improvement rate in the trial diet group being significantly higher than that in the control group (P < 0.05). The results are shown in Table 36.

[0349] Table 35 Volume integral changes (integral value, )

[0350]

[0351] * indicates P < 0.05 for comparison between before and after the test; # indicates P < 0.05 for comparison with the control group.

[0352] Table 36 Improvement of main physical signs

[0353]

[0354] * indicates P < 0.05 compared with the control group.

[0355] 2.3 Impact on human safety indicators

[0356] The participants experienced no adverse reactions or allergic reactions during the trial period, and no significant changes in mental state, sleep, diet, urination, or bowel movements were observed. The results of routine blood, urine, and stool tests, as well as blood biochemical parameters before and after the trial are shown in Table 37. All parameters were within normal range before and after the trial.

[0357] Table 37 Results of blood, urine, stool routine and blood biochemical indexes before and after the tasting

[0358]

[0359] 3 Summary

[0360] 101 subjects who met the requirements were randomly divided into a trial group and a control group; the effective number of cases was 100, with 50 in the trial group and 50 in the control group. The trial group took the composition of the present invention for 30 days as required, and the control group served as a blank control. The improvement rates of the main clinical symptoms and signs in the trial group were 18.00% and 84.00%, respectively, and the improvement rates of the main clinical symptoms and signs in the control group were 0.06% and 38.00%, respectively. There were significant differences between the two groups (P<0.05); the main clinical symptom scores and sign scores in the trial group decreased after the trial, and the difference was statistically significant compared with the control group before and after the trial (P<0.05), indicating that the composition of the present invention has the function of clearing the throat. The blood routine, urine routine, stool routine and blood biochemical indicators of the two groups before and after the trial were all within the normal range, and no allergies or other adverse reactions were observed during the trial. This shows that this product has no adverse effects on the health of the subjects. According to the criteria of the throat clearing function evaluation method (National Food and Drug Administration

[2012] No. 107), the composition of the present invention has the throat clearing function. BRIEF DESCRIPTION OF THE DRAWINGS

[0361] Figure 1 This is a specific chromatogram of the chlorogenic acid high performance liquid chromatography methodology investigation of the present invention. DETAILED DESCRIPTION

[0362] Example 1:

[0363] 360g honeysuckle; 360g Scrophularia ningpoensis; 300g green fruit; 180g Platycodon grandiflorum; 180g Phyllanthus emblica; 30g Momordica grosvenori; 1.2g menthol. (1) Menthol grinding: Weigh menthol and grind it, pass through a 100-mesh sieve, and set aside;

[0364] (2) Extraction: Take honeysuckle, Scrophularia, Cibotrys, Platycodon, Emblica, and Momordica grosvenori according to the formula ratio, add water and extract twice, each time for 1 hour, add 12 times the amount of water for the first time and 10 times the amount of water for the second time, filter the extract and set aside:

[0365] (3) Concentration: The extract is concentrated under reduced pressure (temperature 60-80°C, vacuum degree 0.04-0.08 MPa) to a clear paste with a relative density of 1.08-1.12 (60°C); an appropriate amount of ethanol is added to the clear paste to make the alcohol content 70%, stirred evenly, and allowed to stand at 2-10°C for 48 hours; filtered, the filtrate is collected, the ethanol is recovered under reduced pressure and concentrated (temperature 60-80°C, vacuum degree 0.04-0.08 MPa) to a thick paste with a relative density of 1.20-1.30 (60°C);

[0366] (4) Drying: Use vacuum belt drying with a heating plate temperature of 110-120°C and a vacuum degree of 0.08-0.10 MPa to collect the dry paste for later use;

[0367] (5) Grinding: Grind the dry paste obtained by vacuum belt drying, pass it through a 100-mesh sieve, and set aside. The dry paste powder and menthol powder together constitute the active component of the Chinese medicine preparation and Chinese medicine composition of the present invention;

[0368] (6) Granulation: Place the above dry powder in a mixer, add 300g of lactose, 164g of dextrin and 60g of erythritol, mix well, add 80-90% ethanol to make a soft material, granulate with a 16-mesh sieve, and dry at below 60°C.

[0369] (7) Granulation: Take the dried granules and place them in a crusher, and sieve them through a 1.5 mm conical sieve;

[0370] (8) Total mixing: Add menthol powder, 4 g of magnesium stearate, and 0.4 g of sucralose to the granules after granulation and mix them evenly in a mixer at 8 rpm for 20 minutes;

[0371] (9) Tablet pressing: Tablets are pressed into tablets, and each tablet weighs 0.8 g.

[0372] Example 2

[0373] Determination of total saponin content in the tablets of the present invention

[0374] Ultraviolet spectrophotometry was used to determine the total saponins contained in the samples. The specific contents are as follows:

[0375] Detection method

[0376] 1 reagent

[0377] 1.1Amberlite-XAD-2 macroporous resin.

[0378] 1.2 Ethanol of analytical grade.

[0379] 1.3 For neutral alumina chromatography, 100-200 mesh.

[0380] 1.4 Ginsenoside Re standard: purchased from China Food and Drug Inspection Institute.

[0381] 1.5 Perchloric acid analytical grade.

[0382] 1.6 Glacial acetic acid analytical grade.

[0383] 1.7 Vanillin solution Weigh 5 g of vanillin, dissolve it in glacial acetic acid and dilute to 100 mL.

[0384] 1.8 Preparation of ginsenoside Re standard solution: Accurately weigh 0.020 g of ginsenoside Re standard solution, dissolve it in methanol and make up to 10.0 mL, i.e., each mL contains 2.0 mg of ginsenoside Re.

[0385] 2 Instruments

[0386] 2.1 UV-Visible Spectrophotometer

[0387] 2.2 Glass chromatography column

[0388] 3 Test steps

[0389] 3.1 Sample processing

[0390] Take 10 tablets of the present invention, grind them into powder, mix them evenly, take about 1.5 g, accurately weigh them, place them in a 100 mL volumetric flask, add 70 mL of water, ultrasonicate for 40 minutes, then dilute to 100 mL with water, shake well, let it stand, and draw 1.0 mL of the supernatant for column chromatography.

[0391] 3.2 Column chromatography

[0392] Use a glass chromatography column (1.5 cm inner diameter) filled with 3 cm of Amberlite-XAD-2 macroporous resin and topped with 1 cm of neutral alumina. First, wash the column with 25 mL of 70% ethanol, discard the eluent, then wash the column with 25 mL of water, discard the eluent, accurately add 1.0 mL of the treated sample solution, wash the column with 25 mL of water, discard the eluent, and elute the ginsenosides with 25 mL of 70% ethanol. Collect the eluent in an evaporating dish and evaporate to dryness in a 60°C water bath. This will be used for color development.

[0393] 3.3 Color Rendering

[0394] Accurately add 0.2 mL of 5% vanillin glacial acetic acid solution to the above-mentioned evaporated evaporating dish, rotate the evaporating dish to dissolve the residue, then add 0.8 mL of perchloric acid, mix well, and transfer to a 5 mL stoppered graduated centrifuge tube. Heat in a 60°C water bath for 10 minutes, remove from the tube, cool in an ice bath, accurately add 5.0 mL of glacial acetic acid, shake well, and perform colorimetric determination at a wavelength of 560 nm using a 1 cm colorimetric cell together with a standard tube.

[0395] 3.4 Standard tube

[0396] Pipette 50 μL of ginsenoside Re standard solution (2.0 mg / mL) into an evaporating dish and evaporate in a water bath (below 60°C). The following procedures, starting from "3.2 Column Chromatography...", are the same as for the sample, and the absorbance value is measured.

[0397] 4 Result calculation

[0398]

[0399] Where:

[0400] X: total saponin content in the sample (calculated as ginsenoside Re), g / 100g;

[0401] A1: absorbance value of the liquid being tested;

[0402] A2: absorbance value of the standard solution;

[0403] C: Amount of ginsenoside Re in the standard tube; μg;

[0404] V1: sample volume; mL;

[0405] V2: sample loading volume; mL;

[0406] m: sample mass; g.

[0407] Example 3

[0408] Determination of chlorogenic acid content in the tablets of the present invention

[0409] High performance liquid chromatography was used to determine the characteristic component chlorogenic acid contained in the sample. The specific contents are as follows:

[0410] Detection method

[0411] 1 reagent

[0412] 1.1 Acetonitrile chromatographically pure

[0413] 1.2 Acetic acid analytical grade

[0414] 1.3 Methanol analytical grade

[0415] 1.4 Water Ultrapure Water

[0416] 1.5 Chlorogenic acid standard: purchased from China Food and Drug Inspection Institute for content determination.

[0417] 1.6 Preparation of Chlorogenic Acid Standard Solution: Weigh 0.02 g (accurate to 0.0001 g) of Chlorogenic Acid Standard into a 100.0 mL volumetric flask, dissolve it with mobile phase, dilute to volume, and shake well. This means each mL contains 0.2 mg of Chlorogenic Acid.

[0418] 2 Instruments

[0419] 2.1 High-performance liquid chromatography

[0420] 2.2 Ultrasonic Cleaner

[0421] 3 Test steps

[0422] 3.1 Sample processing

[0423] Take 10 tablets of the present invention, grind them into powder, mix them evenly, weigh 0.2 g (accurate to 0.0001 g), add 20 mL of 70% methanol to a 25.0 mL volumetric flask, ultrasonicate for 30 minutes, dilute to the mark with 70% methanol, shake well, filter through a 0.45 μm microporous membrane, and analyze the filtrate by liquid chromatography.

[0424] 3.2 Preparation of standard curve

[0425] The chlorogenic acid standard solutions were drawn separately, diluted with mobile phase and made up to the concentrations of 2.00, 10.0, 20.0, 40.0 and 80.0 μg / mL standard series in volumetric flasks.

[0426] 3.3 Liquid chromatography conditions

[0427] 3.3.1 Chromatographic column: ODSC18 column, 250 mm × 4.6 mm, 5 μm;

[0428] 3.3.2 Mobile phase: 0.5% acetic acid solution: acetonitrile = 9:1;

[0429] 3.3.3 Flow rate: 1.0 mL / min;

[0430] 3.3.4 Column temperature: 35°C;

[0431] 3.3.5 Detection wavelength: 327nm;

[0432] 3.3.6 Injection volume: 10 μL;

[0433] 3.4 Sample determination

[0434] Inject the standard solution and sample solution into the liquid chromatograph and quantify by comparing the peak area of ​​the sample with that of the standard.

[0435] 4 Result calculation

[0436]

[0437] Where:

[0438] X—chlorogenic acid content in the sample, g / 100g;

[0439] c—the concentration of chlorogenic acid in the injection solution obtained from the standard curve, μg / mL;

[0440] V—sample constant volume, mL;

[0441] m—sample mass, g.

Claims

1. A Chinese medicine preparation having the function of clearing the throat and moistening the throat, characterized in that: The Chinese medicine preparation is prepared from the following raw materials in parts by weight: 36 parts of honeysuckle; 36 parts of Scrophularia; 30 parts of green fruit; 18 parts of Platycodon; 18 parts of Emblica; 3 parts of Momordica grosvenori; 0.12 parts of menthol; 30 parts of lactose; 16.4 parts of dextrin; 6 parts of erythritol; 0.4 parts of magnesium stearate; and 0.04 parts of sucralose. The dosage form of the Chinese medicine preparation is tablets, and each 100 grams of the tablets contains a total amount of not less than 400 mg of total saponins and not less than 240 mg of chlorogenic acid.

2. The Chinese medicine preparation having the function of clearing the throat and moistening the throat according to claim 1, wherein The total saponins are determined by UV-visible spectrophotometry, which includes the following steps: 1) Preparation of standard solution: Weigh 0.020 g of ginsenoside Re standard, dissolve it in methanol and dilute to 10.0 mL, i.e., 2.0 mg of ginsenoside Re per mL; 2) Preparation of test solution: Take 10 tablets, grind them into powder, take 1.5 g, place them in a 100 mL volumetric flask, add 70 mL of water, sonicate for 40 minutes, then dilute to 100 mL with water, let stand, and the supernatant is used as the test solution; 3) Column chromatography: Use a glass chromatography column filled with 3 cm of Amberlite-XAD-2 macroporous resin and topped with 1 cm of neutral alumina. Wash the column with 25 mL of 70% ethanol, discard the eluent, then wash the column with 25 mL of water, discard the eluent, add 1.0 mL of the test solution, wash the column with 25 mL of water, discard the eluent, and elute the ginsenosides with 25 mL of 70% ethanol. Collect the eluent in an evaporating dish and evaporate to dryness in a 60°C water bath. 4) Color reaction: Add 0.2 mL of 5% vanillin glacial acetic acid solution to the dried evaporating dish to dissolve the residue. Then add 0.8 mL of perchloric acid and transfer to a 5 mL graduated centrifuge tube with a stopper. Heat in a 60°C water bath for 10 minutes. After cooling in an ice bath, add 5.0 mL of glacial acetic acid to obtain the product. 5) Determination The same method as steps 3) and 4) is used for column chromatography and color reaction of the standard solution. The absorbance is measured at 560 nm by UV-visible spectrophotometry to calculate the total saponin content in the test sample.

3. The Chinese medicine preparation having the function of clearing the throat and moistening the throat according to claim 1, wherein The chlorogenic acid is determined by high performance liquid chromatography, and the liquid chromatography conditions are as follows: Chromatographic column: ODS C18 column, 250 mm × 4.6 mm, 5 μm; Mobile phase: 0.5% acetic acid solution: acetonitrile = 9:1 (V / V); Flow rate: 1.0 mL / min; Column temperature: 35°C; Detection wavelength: 327nm; Injection volume: 10 μL.

4. The Chinese medicine preparation having the function of clearing the throat and moistening the throat according to claim 3, characterized in that The chlorogenic acid is determined by high performance liquid chromatography, which also includes the following steps: Preparation of chlorogenic acid standard solution: Weigh 0.02 g of chlorogenic acid standard into a 100.0 mL volumetric flask, dissolve it with mobile phase and make up to volume, i.e., 0.2 mg of chlorogenic acid per mL; Preparation of sample solution: Take 10 tablets, grind them into powder, weigh 0.2 g, add 20 mL of 70% methanol to a 25.0 mL volumetric flask, sonicate for 30 minutes, dilute to volume with 70% methanol, filter through a 0.45 μm microporous membrane, and use the filtrate as the sample solution; Preparation of standard curve: Take the chlorogenic acid standard solution, dilute it with mobile phase and prepare a series of standard solutions with concentrations of 2.00, 10.0, 20.0, 40.0, and 80.0 μg / mL in a volumetric flask for HPLC determination. Draw the standard curve with the measured chromatographic peak area as the ordinate and the corresponding standard solution concentration as the abscissa; Sample determination: inject the standard solution and sample solution into the liquid chromatograph, determine, and calculate the chlorogenic acid content in the sample.

5. The Chinese medicine preparation having the function of clearing the throat and moistening the throat according to claim 1, wherein The preparation method of the tablet comprises the following steps: (1) Menthol grinding: Weigh menthol, grind it, pass it through a 100-mesh sieve, and set aside; (2) Extraction: Take honeysuckle, Scrophularia, Cibotrys, Platycodon, Emblica, and Momordica grosvenori according to the formula ratio, add water and extract twice, each time for 1 hour, add 12 times the amount of water for the first time and 10 times the amount of water for the second time, filter the extract and set aside: (3) Concentration: The extract is concentrated under reduced pressure at a temperature of 60-80°C and a vacuum degree of 0.04-0.08 MPa to form a clear paste with a relative density of 1.08-1.12; an appropriate amount of ethanol is added to the clear paste to make the alcohol content 70%, the mixture is stirred evenly, and the mixture is allowed to stand at 2-10°C for 48 hours; the mixture is filtered, the filtrate is collected, the ethanol is recovered under reduced pressure, and the mixture is concentrated at a temperature of 60-80°C and a vacuum degree of 0.04-0.08 MPa to form a thick paste with a relative density of 1.20-1.30; (4) Drying: Use vacuum belt drying with a heating plate temperature of 110-120°C and a vacuum degree of 0.08-0.10 MPa to collect the dry paste for later use; (5) Grinding: Grind the dry paste obtained by vacuum belt drying, pass it through a 100-mesh sieve, and set aside; (6) Granulation: Place the above dry powder in a mixer, add lactose, dextrin and erythritol, mix well, add 80-90% ethanol to make a soft material, granulate with a 16-mesh sieve, and dry at below 60°C. (7) Granulation: Take the dried granules and place them in a crusher, and sieve them through a 1.5 mm conical sieve; (8) Total mixing: Add menthol powder, magnesium stearate, and sucralose to the granules after granulation and mix them evenly in a mixer at 8 rpm for 20 minutes; (9) Tablet pressing: Tablets are pressed into tablets, and each tablet weighs 0.8 g.

6. Use of the traditional Chinese medicine preparation with throat clearing and moistening function according to any one of claims 1 to 5 in the preparation of throat clearing and moistening medicines or health foods.

7. The use according to claim 6, characterized in that The medicine or health food is a medicine or health food for treating or improving chronic pharyngitis.

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