CN106620498A - Preparation method, quality control and use of traditional Chinese medicine composition for treating postinfectious cough - Google Patents

Abstract

The invention provides a traditional Chinese medicine composition for treating cough after a cold, which is composed of the following raw materials in parts by weight: 6-20 grams of Clematis, 6-20 grams of Curcuma, 3-10 grams of Shichangpu, and 3-6 grams of Peppermint gram. Preparation method: Clematis clematis, peppermint, calamus and turmeric medicinal materials, add 6-12 times the total amount of medicinal materials with water, extract volatile oil by steam distillation for 5-9 hours, collect volatile oil, add Tween-80 to solubilize or ethanol Dissolve and set aside; filter the extract by steam distillation to obtain oil extraction mother liquor and filter residue; add 6-10 times water to the filter residue and decoct 1-3 times, each time for 1-3 hours, combine the decoction and oil extraction mother liquor, and After the supernatant is concentrated, add 75-95% ethanol to make the alcohol content reach 65-85%, take it out after refrigerating and standing still, filter, and recover the ethanol from the filtrate under reduced pressure until it has no alcohol smell; add the solubilized volatile oil and conventional medicinal The excipients are made into suitable dosage forms by conventional preparation methods in traditional Chinese medicine pharmacy. The invention also provides the application of the traditional Chinese medicine composition in the preparation of medicines for treating infectious cough and various chronic coughs.

Description

Preparation method, quality control and use of a traditional Chinese medicine composition for treating post-cold cough

technical field

The invention belongs to the field of traditional Chinese medicines and relates to a traditional Chinese medicine composition, in particular to a traditional Chinese medicine composition for treating post-cold cough and its preparation method and application.

Background technique

Cold, also known as acute upper respiratory tract infection, is one of the most common infectious diseases in humans. Adults have an average of 2-4 colds per year. Acute cough is the most common symptom associated with cold and the most common symptom in primary healthcare institutions. one.

According to statistics, about 11%-25% of patients experience post-influenza cough (PIC) every year. The cough is still protracted, mostly manifested as irritating dry cough or coughing a small amount of white mucus sputum, which can last for several weeks, or even longer. Strong negative impact, while traditional Chinese medicine treatment of this disease has certain advantages.

PIC belongs to the category of "exogenous cough disease" in traditional Chinese medicine. Traditional Chinese medicine believes that the lung governs qi, governs breathing, connects the airway, opens to the nose, and the throat is its portal. It is not resistant to cold and heat, and is easily invaded by evils. Exogenous pathogenic factors of the six evils, led by wind pathogens, may be mixed with cold, heat, dryness, dampness, etc., which cannot be dispersed in time, or miscellaneous cough and phlegm drugs, or blind use of bitter cold and heat-clearing drugs, or astringent lungs Misuse of astringent medicines can cause exogenous pathogens to linger, and the lungs lie in the lungs, and the lungs fail to ventilate and descend, leading to coughing. Therefore, one of the TCM syndrome characteristics of cough after a cold is that the external pathogens are not exhausted.

Another feature of PIC is love between righteousness and evil, and lingering illness. Cough after a cold does not heal for a long time, and can turn into internal injury cough. "Su Wen·Cough Theory" pointed out that "all internal organs and six internal organs make people cough, not only the lung." In addition to lung cough, there are also heart cough, liver cough, spleen cough, kidney cough, etc. At this time, the cough is long and sick. The deeper the bit, the more difficult it is to treat. Guan Qiuhong et al. believe that inappropriate tonification is also a common cause of cough after a cold. Over-supplementation or improper diet can internally injure the spleen and stomach, water dampness stops internally, and is stored in the lungs. Some people think that this disease is mostly caused by wind-heat invading the lungs, violent rage and liver damage, prolonged emotional damage, abnormal qi mechanism, relapse of exogenous pathogens, transformation of heat and blazing body fluid; caused by the throat. It is also believed that the disease is caused by stagnation of liver qi, dysfunction of the spleen, and weak stomach qi, so that body fluid cannot go up to the throat. There are also many doctors who discuss treatment from the perspective of pathological products, advocate treatment from phlegm and blood stasis, and believe that "phlegm" and "stasis" are the most common pathological factors in the pathogenesis of chronic pharyngitis, and phlegm and blood stasis coexist. Some studies have divided PIC into 5 types: the treatment of wind-evil love lung type is mainly to eliminate pathogens, the treatment of cold drink subduing the lung type is to warm the lungs and transform drink to dispel cold, and the treatment of phlegm-heat accumulation in the lung type is mainly to clear the lungs and resolve phlegm. Liver-fire invading the lungs clears the lungs, relieves coughs, softens the liver and relieves urgency, while the type of deficiency of the lung-spleen spleen mainly nourishes the lungs and spleen. "Traditional Chinese Medicine Clinical Diagnosis and Treatment Terminology and Syndrome Part" divides post-cold cough into three types: yin deficiency and internal heat type, qi deficiency and phlegm dampness type, and liver depression and qi stagnation type.

The traditional Chinese medicine composition of the present invention is an effective empirical prescription derived from clinical practice. A large number of clinical cases have been accumulated in long-term clinical practice. It is a small compound prescription composed of four traditional Chinese medicines. quality control and process research. in:

Clematis: property pungent, Cheng, warm. Return bladder channel. Have dispelling rheumatism, dredge meridian effect, be used for rheumatism arthralgia, numbness of limbs, spasms of muscles and vessels, dysflexion and extension. For this reason, the monarch drug in this prescription, due to the rheumatism-dispelling effect of Clematis, is very symptomatic for the most common type of wind-evil attacking the lungs in post-cold cough, and plays the role of dispersing wind and promoting lungs.

Herba Acorus: property is pungent, bitter, warm. GUIXIN, Stomach Meridian. Have one's ideas straightened out expel phlegm, refresh the mind and improve intelligence, eliminate dampness and whet the appetite. For stupor epilepsy, forgetful insomnia, tinnitus and deafness, gastral cavity ruffian is not hungry, and dysentery. As an adjuvant in this prescription, Shichangpu has the effect of reducing phlegm and relieving throat in the treatment of cough after a cold.

Radix Curcumae: acrid, bitter, cold in nature. Return liver, heart, lung meridian. It has the effects of promoting blood circulation and relieving pain, promoting qi circulation and relieving depression, clearing away heart-fire and cooling blood, promoting gallbladder and reducing jaundice. For thoracic hypochondrium tingling pain, obstruction of the chest and heartache. This is the ministerial medicine in the side. In the treatment of cough due to liver depression and qi stagnation, turmeric can play a very good role in promoting qi and relieving depression.

Peppermint: pungent in nature, cool. Return lung, Liver Channel. It has the effects of dispersing wind-heat, clearing the head, relieving the throat, clearing the rash, soothing the liver and promoting qi. It is used for wind-heat and cold, the beginning of wind-temperature, headache, red eyes, sore throat, aphtha, rubella, measles, chest and hypochondrium, and it is the medicine in this prescription.

Contents of the invention

The object of the present invention is to provide a kind of Chinese medicine composition;

The object of the present invention is also to provide a preparation method of the composition;

The object of the present invention is also to provide the purposes of this traditional Chinese medicine composition;

The object of the present invention is also to provide a quality control method of the traditional Chinese medicine composition.

The purpose of the present invention is achieved in the following manner:

The traditional Chinese medicine composition for treating post-cold cough is composed of raw materials in the following weight ratios: 6-20 grams of Clematis, 6-20 grams of Curcuma, 3-10 grams of Shichangpu, and 3-10 grams of Peppermint.

The above raw materials, Clematis chinensis Osbeck, Clematis hexapetala Pall. or Clematis manshurica Rupr. dry roots and rhizomes of Clematis chinensis Osbeck of Ranunculaceae; Dry roots of Curcuma wenyujin Y.H.Chen et C.Ling, Curcuma longa L., Curcuma kwangsiensis S.G.Lee et C.F.Liang or Curcuma phaeocaulis Val. The first two are commonly called "warm turmeric" and "yellow silk turmeric", and the rest are commonly called "osmanthus turmeric" or "green turmeric" according to their different characters; Acorus tatarinowii Schott is derived from the dried rhizome. Mint is derived from the dry aerial part of the mint Metha haplocalyx Briq.

The traditional Chinese medicine composition is preferably composed of raw materials in the following weight ratio: 10 grams of Clematis, 10 grams of Curcuma, 6 grams of Shichangpu, and 6 grams of Peppermint.

The traditional Chinese medicine composition is preferably composed of raw materials in the following weight ratios: 6 grams of Clematis, 15 grams of Curcuma, 6 grams of Shichangpu, and 9 grams of Peppermint.

The traditional Chinese medicine composition is preferably composed of raw materials in the following weight ratio: 12 grams of Clematis, 10 grams of Curcuma, 6 grams of Shichangpu, and 6 grams of Peppermint.

The traditional Chinese medicine composition is preferably composed of raw materials in the following weight ratios: 15 grams of Clematis clematis, 12 grams of curcuma, 9 grams of Shichangpu, and 3 grams of peppermint.

The traditional Chinese medicine composition is preferably composed of raw materials in the following weight ratios: 20 grams of Clematis, 15 grams of Curcuma, 3 grams of Shichangpu, and 9 grams of Peppermint.

The preparation method of the above-mentioned Chinese medicine composition is as follows: Clematis clematis, mint, calamus and turmeric medicinal materials, adding 6-12 times the water of the total amount of medicinal materials, extracting volatile oil by steam distillation for 5-9 hours, collecting volatile oil, adding Tween Solubilize at -80, set aside; filter the steam distillation extract to obtain the oil mother liquor and filter residue; add 6-10 times the water to the filter residue and decoct 1-3 times, 1-3 hours each time, combine the decoction and oil extraction Mother liquor, after concentrating the supernatant, add 75-95% ethanol to make the alcohol content reach 65-85%, take it out after refrigerating and standing still, filter, and recover the ethanol from the filtrate under reduced pressure until it has no alcohol smell; add the solubilized volatile oil and Conventional pharmaceutical excipients are made into suitable dosage forms by conventional preparation methods in traditional Chinese medicine pharmacy.

The above preparation method is preferably: Clematis clematis, peppermint, calamus and turmeric medicinal materials, adding 12 times the total amount of medicinal materials, extracting volatile oil by steam distillation for 7 hours, collecting volatile oil, filtering the mother liquor, adding Tween-80 for solubilization , and set aside; the steam distillation extract was filtered to obtain the oil extraction mother liquor and the filter residue; the filter residue was added to 10 times of water and decocted for 3 times, each time for 2 hours, the decoction and the oil extraction mother liquor were combined, and the supernatant was concentrated and then added with 95% Ethanol makes the alcohol content reach 65%, take it out after refrigerating and standing for 15 hours, filter, and recover the ethanol from the filtrate under reduced pressure until it has no alcohol smell; add the solubilized volatile oil and conventional pharmaceutical excipients, and adopt the conventional preparation method in traditional Chinese medicine pharmacy , made into granules, oral tablets, buccal tablets, dispersible tablets, hard capsules, soft capsules, oral liquids, sprays, powder sprays.

The preferred dosage form of the Chinese medicine composition of the present invention is oral liquid, and its preparation method is: Clematis clematis, peppermint, Shichangpu and turmeric medicinal materials, adding 12 times of water of the total amount of medicinal materials, extracting volatile oil with steam distillation for 7 hours, collecting volatile oil , add Tween-80 to solubilize, and set aside; filter the steam distillation extract to obtain oil extraction mother liquor and filter residue; add 10 times water to decoct the filter residue 3 times, each time for 2 hours, combine the decoction and oil extraction mother liquor, After concentrating the supernatant, add 95% ethanol to make the alcohol content reach 65%, take it out after refrigerating and standing for 15 hours, filter, and recover the ethanol from the filtrate under reduced pressure until it has no alcohol taste; add the solubilized volatile oil, simple syrup and stevioside And sodium benzoate, constant volume, sub-packed in oral liquid bottles, to get.

The Chinese medicine composition of the present invention can also be prepared by the following method: Clematis clematis, peppermint, calamus and turmeric medicinal materials are added to reflux extraction with 50-80% ethanol 5-15 times the total amount of medicinal materials, and the extraction times are 1-4 times. Each time for 0.5-4 hours, the alcohol extracts are combined, filtered, the filtrate is concentrated, conventional pharmaceutical excipients are added, and a suitable dosage form is prepared by adopting a conventional preparation method in traditional Chinese medicine pharmacy.

The above preparation method is preferably: Clematis clematis, mint, calamus and turmeric medicinal materials, add 70% ethanol to reflux extraction twice, each time for 3 hours, combine the alcohol extracts, filter, concentrate, add conventional pharmaceutical excipients, use traditional Chinese medicine The conventional preparation methods in pharmacy are made into granules, oral tablets, buccal tablets, dispersible tablets, hard capsules, soft capsules, oral liquids, sprays, and powder sprays.

The preferred dosage form of the traditional Chinese medicine composition of the present invention is oral liquid, and its preparation method is: Clematis clematis, peppermint, calamus and turmeric medicinal materials, add 70% ethanol and reflux extraction twice, each time for 3 hours, the ethanol extracts are combined, filtered , concentrate, add simple syrup, stevioside and sodium benzoate, dilute to volume, pack in oral liquid bottles, and get ready.

The Chinese medicine composition of the present invention can also be prepared by the following method: Clematis clematis, mint, calamus and turmeric medicinal materials, add 6-12 times of water of the total amount of medicinal materials, extract volatile oil for 5-9 hours with steam distillation, collect volatile oil , β-cyclodextrin inclusion complex, to obtain inclusion compound for later use: filter the steam distillation extract to obtain oil mother liquor and filter residue; add 6-10 times of water to the filter residue and decoct 1-3 times, 1-3 hours each time , combine the decoction liquid and oil extraction mother liquor, concentrate the supernatant, add 75-95% ethanol to make the alcohol content reach 65-85%, take it out after refrigerating and standing still, filter, and recover the ethanol from the filtrate under reduced pressure until it has no alcohol smell , and then continue to concentrate to a thick paste, dry, and mix the obtained dried product with the volatile oil clathrate evenly, add conventional pharmaceutical excipients, and adopt the conventional preparation method in traditional Chinese medicine pharmacy to make a suitable dosage form.

The above preparation method is preferably: Clematis clematis, peppermint, calamus and turmeric medicinal materials, adding 12 times the total amount of medicinal materials, extracting volatile oil by steam distillation for 7 hours, collecting volatile oil, β-cyclodextrin inclusion, and refrigerating overnight , filter, and dry at low temperature to obtain β-cyclodextrin inclusion compound for later use; filter the steam distillation extract to obtain oil mother liquor and filter residue; add 10 times water to the filter residue and decoct 3 times, 2 hours each time, Combine the decoction liquid and the oil extraction mother liquid, concentrate the supernatant, add 95% ethanol to make the alcohol content reach 65%, refrigerate and stand for 15 hours, take it out, filter, and recover the ethanol from the filtrate under reduced pressure until there is no alcohol smell, and then continue to concentrate To a thick paste, dry, and mix the obtained dried product with the volatile oil inclusion compound evenly, add conventional pharmaceutical excipients, and adopt conventional preparation methods in traditional Chinese medicine pharmacy to make granules, oral tablets, buccal tablets, dispersible tablets, and hard capsules Agents, soft capsules, oral liquids, sprays, powder sprays.

The preferred dosage form of the Chinese medicine composition of the present invention also has buccal tablets, and its preparation method is: Clematis clematis, mint, calamus and turmeric medicinal materials, add 12 times of water of the total amount of medicinal materials, extract volatile oil with steam distillation for 7 hours, Collect volatile oil, β-cyclodextrin inclusion complex, refrigerate overnight, filter, and dry at low temperature to obtain β-cyclodextrin inclusion complex for later use; filter the steam distillation extract to obtain oil mother liquor and filter residue; add the filter residue Decoct with 10 times water for 3 times, 2 hours each time, combine the decoction liquid and the mother liquid for extracting oil, concentrate the supernatant, add 95% ethanol to make the alcohol content reach 65%, refrigerate and stand for 15 hours, take it out, filter, The filtrate is decompressed to recover ethanol until it has no alcohol smell, then continue to concentrate to a thick paste, dry, and mix the obtained dried product with volatile oil clathrate evenly, add dextrin, sucrose powder, stevioside, essence, wet granulation, and granulate after drying , add 0.5% magnesium stearate, mix evenly, and compress into buccal tablets.

The quality control method of the traditional Chinese medicine composition of the present invention includes one or more of the following identification methods and/or content determination:

Identification:

Take 2-10mL of the content of the traditional Chinese medicine composition preparation of the present invention and place it in a round-bottomed sesame cake, add 2mol/L HCl30-100mL, reflux in a water bath for 1-3 hours, put it in a separatory funnel after cooling, add ethyl acetate Extract 1-3 times, 15-30mL each time, combine the extracts and evaporate to dryness under pressure, dilute to 5-10mL with methanol, and use it as the test solution. Take another oleanolic acid and hedera saponin reference substance plus methanol to make each 1mL containing 1mg, as the reference solution. Test according to thin-layer chromatography ("Chinese Pharmacopoeia" 2010 edition appendix VI B), draw each 1-5 μ l of the above-mentioned two kinds of solutions, point respectively on the same silica gel G thin-layer plate, add petroleum ether-ethyl acetate-formic acid (5 -20∶1-10∶0.1-1) as the developing agent, place the thin-layer plate in the developing cylinder and pre-saturate for 30 minutes, develop, take out, dry in the air, spray with 10% sulfuric acid ethanol solution, and heat at 105°C until the spots develop color clear. In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference product.

Take 2-10mL of the content of the traditional Chinese medicine composition preparation of the present invention and place it in a separatory funnel, add petroleum ether to extract 1-3 times, 5-15mL each time, combine ether, evaporate to dryness under reduced pressure, add 2-10mL methanol to constant volume , as the test solution. Another mint reference medicinal material was taken, crushed through a 40-mesh sieve, 0.5 g was added to 5-15 mL of petroleum ether and ultrasonicated for 30 minutes, filtered, the filtrate was evaporated to dryness and the volume was adjusted to 1 mL, as the reference medicinal material solution. Another menthol reference substance plus methanol made per 1ml containing 2mg, as the reference substance solution. Test according to thin-layer chromatography (appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 1-5 μ l of each of the above two solutions, and place them on the same silica gel G thin-layer plate respectively, and use toluene-ethyl acetate (5-20: 1-5) is the developing agent. After pre-saturation for 30 minutes, develop, take out, dry in the air, spray with 2% vanillin sulfuric acid-ethanol (1:4) color developer, and heat at 105°C until the spots are clearly colored. In the chromatogram of the test product, there are spots of the same color at the positions corresponding to the chromatogram of the reference substance and the reference medicinal material.

Take 2-10mL of the content of the traditional Chinese medicine composition preparation of the present invention, add 25-50mL of absolute ethanol and sonicate for 15-30min, filter, recover the solvent under reduced pressure, and dissolve the residue with 1mL of ethanol as the test solution. Take another curcuma reference medicinal material, crush it through a 40-mesh sieve, take 0.5-2 g, add 5-50 mL of ethanol and sonicate for 30 minutes, filter, and dilute to 1 mL as the reference medicinal material solution. According to the test of thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 1-5 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and use petroleum ether (60-90 ° C)-acetic acid Ethyl ester (10-30:5-10) was used as the developing agent. After pre-saturation for 30 minutes, develop, take out and dry in the air, spray with 10% concentrated sulfuric acid ethanol color developer, and heat at 105°C until the spots are clearly colored. In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the control medicinal material.

Take 2-10mL of the content of the traditional Chinese medicine composition preparation of the present invention and place it in a separatory funnel, add petroleum ether to extract 1-3 times, 5-15mL each time, combine ether, evaporate to dryness under reduced pressure, add 2-10mL methanol to constant volume , as the test solution. Another reference medicinal material, Shichangpu, was crushed through a 40-mesh sieve, and 0.5-2 g was taken, and 10-40 mL of ether was added for ultrasonication for 30 minutes. After filtration, the filtrate was evaporated to dryness and the volume was adjusted to 1 mL, as the test solution. Another α-asarone reference substance plus methanol prepared to contain 2mg per mL, as the reference substance solution. According to the test of thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 1-5 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and use petroleum ether (60-90 ° C)-acetic acid Ethyl ester (5-20:2-20) was used as the developer, pre-saturated for 30 minutes, developed, taken out, dried in the air, and inspected under an ultraviolet lamp (365nm). In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference substance solution.

Content determination:

Chromatographic conditions: octadecylsilane bonded silica gel as filler; mobile phase is acetonitrile (A)-0.1-0.5% phosphoric acid (B), gradient elution (0-15min: 70%A, 15-20min: 70% →90%A, 20-30min: 90%A, 30-35: 90%→70%); column temperature 30°C; flow rate 1.0ml·min-1, detection wavelength 205nm. Quantification of peak area by external standard method.

Preparation of reference substance solution: Weigh the reference substances of hedera saponin and oleanolic acid dried under reduced pressure at 60°C for 3-5 hours, add methanol to obtain a concentration of 20ug per 1ml.

Preparation of the test solution: Measure 1-5ml of the content of the traditional Chinese medicine composition preparation, add 10-50ml of 2mol/L hydrochloric acid solution, heat and reflux for 1-3 hours, cool immediately, transfer to a separatory funnel, add ethyl acetate to shake Extract 1-3 times, 10-30ml each time, combine the ethyl acetate solution, evaporate to dryness, add methanol to dissolve, transfer to a 5ml measuring bottle, and set to volume.

Determination method: draw respectively 10ul of the reference substance solution and the test solution, inject it into the liquid chromatograph, measure it, and obtain it.

The oral liquid of the traditional Chinese medicine composition of the present invention contains not less than 0.60-1.00 mg of Clematis per milligram in terms of hedera saponin, and not less than 0.10-0.40 mg in terms of oleanolic acid.

The quality control method of the Chinese medicine composition of the present invention is preferably one or more of the following identification methods and/or content determination:

Identification:

Take 2 mL of the Chinese medicine composition preparation of the present invention and place it in a round-bottomed sesame seed cake, add 30 mL of 2mol/L HCl, reflux in a water bath for 2 hours, put it in a separating funnel after cooling, add ethyl acetate to extract 3 times, 15 mL each time , The combined extracts were evaporated to dryness under pressure, and the volume was adjusted to 10 mL with methanol as the test solution. Take another oleanolic acid and hedera saponin reference substance plus methanol to make each 1mL containing 1mg, as the reference solution. According to the thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 5 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and use petroleum ether-ethyl acetate-formic acid (10:3 : 0.1) as the developing agent, the thin-layer plate was placed in a developing cylinder and pre-saturated for 30 minutes, developed, taken out, dried in the air, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored. In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference product.

Take 2 mL of the Chinese medicine composition preparation of the present invention and place it in a separatory funnel, add petroleum ether to extract 3 times, 5 mL each time, combine ether, evaporate to dryness under reduced pressure, add 2 mL of methanol to constant volume, as the test solution. Take mint as a control medicinal material, crush it through a 40-mesh sieve, take 0.5 g and add 10 mL of petroleum ether to sonicate for 30 minutes, filter, evaporate the filtrate to dryness and dilute to 1 mL, and use it as the reference medicinal material solution. Another menthol reference substance plus methanol made per 1ml containing 2mg, as the reference substance solution. According to the thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 2 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and develop with toluene-ethyl acetate (8:18) After pre-saturation for 30 minutes, develop, take out, dry in the air, spray with 2% vanillin sulfuric acid-ethanol (1:4) color developer, heat at 105°C until the spots are clear in color. In the chromatogram of the test product, there are spots of the same color at the positions corresponding to the chromatogram of the reference substance and the reference medicinal material.

Get 2mL of the Chinese medicine composition preparation of the present invention, add 25mL of absolute ethanol and sonicate for 15min, filter, recover the solvent under reduced pressure, and dissolve the residue with 1mL of ethanol as the test solution. Take another curcuma reference medicinal material, crush it through a 40-mesh sieve, take 2 g, add 25 mL of ethanol to ultrasonic for 30 minutes, filter, and dilute to 1 mL as the reference medicinal material solution. According to the thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 1-5 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and use petroleum ether (60-90 ° C)-formic acid Ethyl ester (15:5) was used as the developing agent. After pre-saturation for 30 minutes, develop, take out and dry in the air, spray with 10% concentrated sulfuric acid ethanol color developer, and heat at 105°C until the spots are clearly colored. In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the control medicinal material.

Get 5 mL of the Chinese medicine composition preparation of the present invention and place it in a separatory funnel, add petroleum ether to extract 3 times, 5 mL each time, combine ether, evaporate to dryness under reduced pressure, add 5 mL methanol to constant volume, as the test solution. Another reference medicinal material, Shichangpu, was crushed through a 40-mesh sieve, and 1 g was taken, and 10 mL of ether was added for ultrasonication for 30 minutes. After filtration, the filtrate was evaporated to dryness and the volume was adjusted to 1 mL, as the test solution. Another α-asarone reference substance plus methanol prepared to contain 2mg per mL, as the reference substance solution. According to the test of thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 1-5 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and use petroleum ether (60-90 ° C)-acetic acid Ethyl ester (6:4) was used as the developer, pre-saturated for 30min, developed, taken out, dried in the air, and inspected under ultraviolet light (365nm). In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference substance solution.

Content determination:

Chromatographic conditions: octadecylsilane bonded silica gel as filler; mobile phase is acetonitrile (A)-0.1% phosphoric acid (B), gradient elution (0~15min: 70%A, 15~20min: 70%→90 %A, 20-30min: 90%A, 30-35: 90%→70%); column temperature 30°C; flow rate 1.0ml·min ^-1 , detection wavelength 205nm. Quantification of peak area by external standard method.

Preparation of reference substance solution: Weigh the reference substance of hedera saponin and oleanolic acid dried under reduced pressure at 60°C for 4 hours, add methanol to make the concentration of 20ug per 1ml.

Preparation of the test solution: Measure 1ml of the content of the traditional Chinese medicine composition preparation, add 30ml of 2mol/L hydrochloric acid solution, heat and reflux for 3 hours, cool immediately, transfer to a separatory funnel, add ethyl acetate to shake and extract 3 times, each time 15ml, combined ethyl acetate solution, evaporated to dryness, dissolved in methanol, transferred to a 5ml measuring bottle, and made to volume.

Determination method: draw respectively 10ul of the reference substance solution and the test solution, inject it into the liquid chromatograph, measure it, and obtain it.

The oral liquid of the traditional Chinese medicine composition of the present invention contains not less than 0.70 mg of Clematis per milligram in terms of hedera saponin, and not less than 0.25 mg in terms of oleanolic acid.

The assay method of the above claims, characterized in that methanol can be replaced by 70% ethanol or ethyl acetate.

The content determination method of the above claims, characterized in that the ultrasonic treatment method can be replaced by the reflux extraction method.

The content determination method of the above claims, characterized in that the ultrasonic treatment method can be replaced by the reflux extraction method.

The assay method of the above claims is characterized in that the mobile phase acetonitrile-0.1% phosphoric acid can be replaced by acetonitrile-water.

Application of the traditional Chinese medicine composition in preparing medicine for treating infectious cough.

Application of the traditional Chinese medicine composition in preparing various chronic cough medicines.

The following pharmaceutical experiment examples are used to further illustrate but not limit the present invention.

Experimental Example 1: Is it necessary to soak before oil extraction?

According to the prescription, 170g of the medicinal material was weighed in 2 parts, 10 times of water was added, and the volatile oil was extracted by steam distillation. One part was soaked, and the other part was not soaked. The amount of volatile oil collected after 10 hours is shown in the table below. From Table 1 below, it can be seen that the total amount of volatile oil collected by soaking medicinal materials is 0.56ml, and the total amount of volatile oil collected without soaking is 0.48ml. Therefore, medicinal materials need to be soaked before extracting oil.

Table 1 Effect of soaking or not on the amount of volatile oil

Experimental Example 2: Investigation of Water Adding Multiples

According to the prescription, 170g of medicinal materials were weighed in 4 parts, and 8, 10, 12, 14 times of water were added respectively, and the volatile oil was extracted by steam distillation, and the cumulative amount of volatile oil extracted within 10 hours was observed. Table 2 shows the amount of volatile oil extracted at different times of adding water.

Table 2 Effect of water addition ratio on the amount of volatile oil

From the total amount of volatile oil extracted in the table, the amount of volatile oil extracted by adding 12 times of water without soaking is 0.51ml, which is better than the amount of oil extracted by adding water multiples. After 7 hours of extraction, the amount of volatile oil is about 0.51ml, which is basically the same. Therefore, the optimal process for extracting volatile oil is determined as adding 12 times the amount of medicinal materials in water, without soaking, and the oil extraction time is 7 hours.

Experimental Example 3: Extraction Process Optimization Research

After the volatile oil is extracted from the prescribed medicinal materials, the dregs are decocted in water. Orthogonal design method was used to optimize the decoction process parameters. The transfer rate and ointment rate of hedera saponin and oleanolic acid were used as indicators, and the water extraction process was investigated by selecting the water extraction time, the number of water extraction times, and the multiple of water addition. The results are shown in Table 3 and Table 4.

Table 3 Factor levels of water extraction process

Table2-6 Factors and levels of water extraction of compound radix clematidis sprays

Table 4 Orthogonal design experiment results table

Through the analysis of the data results of the orthogonal experiment, it is found that the factors affecting the transfer rate of helexin in Clematis clematis in the water extraction experiment are in the order of primary and secondary: times > water amount > time, that is, the effect of extraction times on the transfer rate of helexin in Clematis clematis The most significant impact, followed by the amount of water added, the least impact of extraction time. Investigate the changes of the three factors on the three levels, the average value of the three levels of the time factor, the number of times factor and the amount of water added is the largest, which is the optimal condition, so the best process condition is determined as adding water and decocting three times, Every 2 hours, add 10 times the amount of water.

Experimental Example 4: Research on Alcohol Precipitation Process

Orthogonal design method was adopted, and hedera saponin was used as an index, and the concentrated solution concentration, alcohol deposition fraction, and standing time were selected to investigate the alcohol precipitation process. The factor levels are shown in Table 5 and Table 6.

Table 5 Factor levels of alcohol precipitation experiments

Table 6 Orthogonal design experiment results table

Through the analysis of the data results of the orthogonal experiment, it is found that in the alcohol precipitation experiment, the factors affecting the transfer rate of helexin in Clematis clematis are in the order of primary and secondary: concentrated solution concentration > standing time > alcohol precipitation volume fraction, that is, the concentration of the concentrated solution has a significant effect on the transfer rate of Clematis clematis The transfer rate of helexin in Xianzhong had the most significant effect, followed by the standing time, and the alcohol precipitation volume fraction had the least effect. Investigate the changes of the three factors on the 3 levels, the average value of the 2 levels of the concentrated solution concentration factor, the 1 level of the alcohol precipitation volume fraction factor and the 1 level of the standing time is the largest, which is the optimal condition, so the optimum process conditions are determined as The concentrated solution concentration (g/ml) was 1.0, the alcohol concentration was 60%, and the standing time was 12 hours.

Experimental example 5: the choice of flavoring agent and dosage

Since the liquid taste of this traditional Chinese medicine composition is bitter and difficult to swallow, and the tongue has an obvious thick feeling after swallowing, it is necessary to consider adding an appropriate amount of corrective agent to improve the mouthfeel of the preparation and improve the compliance of patients with medication. In the choice of flavoring agent, the taste was slightly worse after adding sucrose, honey, brown sugar, aspartame, etc., and finally stevioside was selected, and the dosage was screened after several people tasted it. In the end, adding 0.33% stevia was chosen as a more suitable prescription. The results are shown in Table 7.

Table 7 Screening results of stevioside dosage

Experimental Example 6: Screening of preservatives

In order to avoid deterioration and rancidity caused by microbial contamination, it is often necessary to add an appropriate amount of preservatives to the extract of traditional Chinese medicine. Sodium benzoate has no toxic and side effects, and has a good taste. It is an ideal preservative for traditional Chinese medicine liquid preparations. According to the usual dosage of sodium benzoate as antiseptic Liu, its dosage is determined to be 0.30% through preliminary tests.

Experimental Example 7: Qualitative identification of the main components contained in Clematis clematis

Take 2 mL of the Chinese medicine composition preparation of the present invention and place it in a round-bottomed sesame seed cake, add 30 mL of 2mol/L HCl, reflux in a water bath for 2 hours, put it in a separating funnel after cooling, add ethyl acetate to extract 3 times, 15 mL each time , The combined extracts were evaporated to dryness under pressure, and the volume was adjusted to 10 mL with methanol as the test solution.

Take another oleanolic acid and hedera saponin reference substance plus methanol to make each 1mL containing 1mg, as the reference solution. According to the thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 5 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and use petroleum ether-ethyl acetate-formic acid (10:3 : 0.1) as the developing agent, the thin-layer plate was placed in a developing cylinder and pre-saturated for 30 minutes, developed, taken out, dried in the air, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored. In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference product.

Experimental Example 8: Identification of main components contained in peppermint

Take 2 mL of the Chinese medicine composition preparation of the present invention and place it in a separatory funnel, add petroleum ether to extract 3 times, 5 mL each time, combine ether, evaporate to dryness under reduced pressure, add 2 mL of methanol to constant volume, as the test solution.

Take mint as a control medicinal material, crush it through a 40-mesh sieve, take 0.5 g and add 10 mL of petroleum ether to sonicate for 30 minutes, filter, evaporate the filtrate to dryness and dilute to 1 mL, and use it as the reference medicinal material solution.

Another menthol reference substance plus methanol made per 1ml containing 2mg, as the reference substance solution. According to the thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 2 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and develop with toluene-ethyl acetate (8:18) After pre-saturation for 30 minutes, develop, take out, dry in the air, spray with 2% vanillin sulfuric acid-ethanol (1:4) color developer, heat at 105°C until the spots are clear in color. In the chromatogram of the test product, there are spots of the same color at the positions corresponding to the chromatogram of the reference substance and the reference medicinal material.

Experimental Example 9: Identification of Main Components in Curcuma

Take 2mL of the Chinese medicine composition preparation of the present invention, add 25mL of absolute ethanol and sonicate for 15min, filter, recover the solvent under reduced pressure, and dissolve the residue with 1mL of ethanol as the test solution.

Take another curcuma reference medicinal material, crush it through a 40-mesh sieve, take 2 g, add 25 mL of ethanol to ultrasonic for 30 minutes, filter, and dilute to 1 mL as the reference medicinal material solution. According to the thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 1-5 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and use petroleum ether (60-90 ° C)-formic acid Ethyl ester (15:5) was used as the developing agent. After pre-saturation for 30 minutes, develop, take out and dry in the air, spray with 10% concentrated sulfuric acid ethanol color developer, and heat at 105°C until the spots are clearly colored. In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the control medicinal material.

Experimental Example 10: Identification of main components contained in Shichangpu

Get 5 mL of the Chinese medicine composition preparation of the present invention and place it in a separatory funnel, add petroleum ether to extract 3 times, 5 mL each time, combine ether, evaporate to dryness under reduced pressure, add 5 mL methanol to constant volume, as the test solution.

Another reference medicinal material, Shichangpu, was crushed through a 40-mesh sieve, and 1 g was taken, and 10 mL of ether was added for ultrasonication for 30 minutes. After filtration, the filtrate was evaporated to dryness and the volume was adjusted to 1 mL, as the test solution.

Another α-asarone reference substance plus methanol prepared to contain 2mg per mL, as the reference substance solution. According to the thin-layer chromatography (Appendix VI B of "Chinese Pharmacopoeia" 2010 edition), draw 1-5 μl of each of the above two solutions, respectively spot on the same silica gel G thin-layer plate, and use petroleum ether (60-90 ℃)-acetic acid Ethyl ester (6:4) was used as the developer, pre-saturated for 30min, developed, taken out, dried in the air, and inspected under ultraviolet light (365nm). In the chromatogram of the test product, there are spots of the same color at the position corresponding to the chromatogram of the reference substance solution.

Experimental Example 11: Determination of Hedera Saponin and Oleanolic Acid Contents in Traditional Chinese Medicine Composition Preparations

Selection of detection wavelength: take an appropriate amount of the mixed reference solution, and conduct ultraviolet scanning at a wavelength of 200-400nm. The three components have characteristic absorption at a wavelength of 205nm, so 205nm is selected as the detection wavelength.

Chromatographic determination conditions: acetonitrile (A)-0.1% phosphoric acid (B), gradient elution (0～15min: 70%A, 15～20min: 70%→90%A, 20～30min: 90%A, 30～35 : 90%→70%); column temperature 30°C; flow rate 1.0ml·min-1; The total shall not be less than 1000.

Linear relationship investigation:

Accurately weigh 5.73 mg of hedera saponin and oleanolic acid reference substance, put it in a 25ml volumetric flask, and set it to volume with chromatographic methanol; Concentrate to the scale, pass through a 0.22 μm organic filter membrane, inject 10 μl into a high-performance liquid chromatograph to measure the peak area according to the above-mentioned chromatographic conditions, use the integral value of the peak area as the ordinate, and use the injection volume of hedera saponin and oleanolic acid respectively (ug) is the abscissa, draws the standard curve, calculates the regression equation, and the results are shown in Table 8.

Table 8 Investigation of the linear relationship between hedera saponin and oleanolic acid

Standard curve: helexin Y=6E+0.6X+40997, R2=0.9999; standard curve: oleanolic acid Y=6E+0.6X+61056, R2=0.9996;

Reproducibility test:

Precisely draw 2ml of the Chinese medicine composition preparation sample of the present invention, parallel 8 parts, add 2mol/L 30ml HCL respectively, heat and reflux for 2h, let cool, transfer to a separatory funnel, extract 5 times with ethyl acetate, combine the extracts, and recover the solvent , dried under reduced pressure, the residue was dissolved in chromatographic methanol, transferred to a constant volume of 10ml, passed through a 0.22μm organic filter membrane, and 10μl was injected into a high performance liquid chromatograph to measure the peak area according to the above chromatographic conditions.

Table 9 Repeatability investigation of hedera saponin and oleanolic acid

Conclusion: The RSD values of hedera saponin and oleanolic acid are less than 5%, indicating that the method has good repeatability.

Precision test:

Precisely draw 10 μL of the above-mentioned reproducibility experiment sample 8 solution, repeat the injection 6 times, and obtain the relative standard deviation RSD of the peak areas of helexin and oleanolic acid<1%. The results are shown in Table 10.

Table 10 Helexin precision test results

Stability test

Precisely draw 10 μL of the above-mentioned reproducibility test sample, inject 1 sample at 0, 2, 4, 6, 10, 12 respectively, and inject 6 samples in total. The measurement results are shown in Table 11. Hederinin and oleanolic acid The peak area was used to test the stability of the sample.

Table 11 Stability test of hedera saponin and oleanolic acid

Conclusion: The peak areas of helexin and oleanolic acid of the same sample are within 12 hours, and the RSD value is less than 1%, indicating that the test solution is stable within 12 hours.

Recovery test:

According to the repeated investigation results, the content of helexin was 0.90 mg/ml; the content of oleanolic acid was 0.40 mg/ml. Therefore, accurately weigh 9.40 mg of hedera saponin and 4.00 mg of oleanolic acid in a 10 ml volumetric flask, constant volume with methanol for chromatography, and fully dissolve them to obtain a mixed standard solution.

Accurately draw 1ml of the above mixed standard solution in the Erlenmeyer flask, parallel 7 parts, add 1ml of the pilot test sample precisely drawn, add 2mol/L 30ml HCL, heat and reflux for 2h, let cool, transfer to a separatory funnel, and extract with ethyl acetate for 5 Once, 15ml each time, combine the extracts, dry the solvent under reduced pressure, dissolve the residue with chromatographic methanol, set the volume to 10ml, pass through a 0.22μm organic filter membrane, inject 10μl into the high performance liquid chromatograph and measure the peak area according to the above chromatographic conditions.

Calculate the sample recovery rate according to the following formula:

Table 12 Recovery rate of hedera saponin and oleanolic acid

Conclusion: the recovery rates of 6 samples are all between 95%-105%, and the RSD values are all less than 5%, which shows that the method is stable and reliable, and can be used for helexin and oleanolic acid in the Chinese medicine composition of the present invention content determination.

The following pharmacodynamic experiment results are used to further illustrate but not limit the present invention.

Experimental Example 11: Experimental observation on the efficacy of cough medicine

1. The purpose of the experiment: to compare the antitussive effects of ammonia water spray through the cough test.

2. Experimental materials

2.1 Drugs under test Process 1, Process 2, and Process 3 of the cough medicine were provided by the Preparation Center of the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, and the contents were all 1.0 g crude drug/ml.

2.2 Animal CD-1 (ICR) mice, 18-22g, provided by Beijing Weitong Lihua Experimental Animal Technology Co., Ltd., the certificate number is SCXK (Beijing) 2012-0001.

2.3 Reagent Ammonia water, Sinopharm Chemical Reagent Co., Ltd., batch number: 20120918.

2.4 Instrument 402A ultrasonic nebulizer, produced by Jiangsu Yuyue Medical Equipment Co., Ltd.

2.5 Animal grouping

Get healthy mice and divide into 10 groups at random according to body weight, be respectively model control group, embodiment 1 drug high, middle and low dose groups, embodiment 2 high, middle and low dose groups, embodiment 3 high, middle and low dose groups Low-dose group, 10 rats in each group, half male and half male.

3. Experimental method

Ammonia water spray was used to induce cough, and the drugs of Example 1, Example 2, and Example 3 were given to mice by intragastric administration of 12.54, 6.27, and 3.14 g of crude drug/kg, respectively, and their antitussive effects were compared through an antitussive test. Each group was intragastrically given the corresponding drug one by one, the volume of intragastric administration was 20ml/kg, and the model control group was intragastrically administered with the same volume of distilled water. One hour after the administration, the mice were put into a 1000ml glass beaker one by one. Cover it, put 40ml of 12.5% ammonia water into the atomizing cup, connect the atomizer and the glass beaker with the mist delivery pipe, turn on the machine with a certain power of moderate air supply and maximum mist delivery, stop spraying for 5 seconds, observe and record the cause The time required for mice to cough (cough latency) and the number of coughs within 2 minutes were compared between groups by t test.

4. Experimental results

Table 13 The effect of ammonia water spray on coughing in mice

Note: Compared with the control group, *P<0.05; **P<0.01

5. Conclusions of the observation experiment on the efficacy of cough medicine

As seen from the test results, the high, middle and low dosage groups of Chinese medicine composition embodiment 1 of the present invention and the high and middle dosage groups of embodiments 2 and 3 all can obviously prolong the cough latency of mice; the high and middle dosage groups of embodiment 1 , low-dose group, the high-dose group of embodiment 2 and the high-dose and middle-dose groups of embodiment 3 can significantly reduce the number of mice coughing, compared with the model control group, there is a significant difference.

detailed description:

Embodiment 1: the preparation of oral liquid

Medicinal materials Clematis 300g, Curcuma 250g, Shichangpu 150g, Peppermint 150g, add 10 times of water, extract volatile oil by steam distillation, collect volatile oil after 8 hours for later use, filter out the mother liquor of oil extraction, add 10 times of medicinal residue to decoct Boil for 1 hour, filter, combine the filtrate with oil extraction mother liquor, and centrifuge at 3500r/min for 20 minutes to remove the precipitate. After concentration, add 95% ethanol to make the alcohol content reach 65%, take it out after refrigerating and standing overnight, filter, and recover the ethanol from the filtrate under reduced pressure, add Tween 80 to solubilize the volatile oil, and mix it with correctives (such as sucrose, honey, wood One or more of sugar alcohol, stevia sugar, etc.), preservatives (such as one or more of sodium sorbate or potassium salt, benzoic acid, etc.) and liquid medicine are mixed evenly, and the volume is set to 1000ml, and the filling is ready.

Embodiment 2: the preparation of common tablet

Medicinal materials Clematis 150g, Curcuma 375g, Shichangpu 150g, Peppermint 225g, add 10 times the amount of water, extract the volatile oil by steam distillation, collect the volatile oil for 7 hours for later use, filter out the mother liquor of oil extraction, add 8 times the amount of medicinal residue 60% ethanol, reflux twice, 1 hour each time, combine the ethanol extracts, recover the ethanol until it has no alcohol smell, after standing overnight, remove the precipitate by suction filtration, concentrate to dryness, and pulverize. The volatile oil is clathrated with β-cyclodextrin, refrigerated overnight, filtered, dried at low temperature, added with dextrin, powdered sugar and magnesium stearate, mixed with extract powder, granulated, and pressed into 1000 tablets.

Embodiment 3: the preparation of spray

Add 225g of medicinal materials Acorus calamus and 75g of mint with 14 times of water, extract the volatile oil by steam distillation, collect the volatile oil for 7 hours for later use, filter out the oil extraction mother liquor, add 10 times of water to the medicinal residue, decoct for 1 hour, and filter the decoction Afterwards, it was combined with the oil extraction mother liquor, and the precipitate was removed by centrifugation at 3500r/min for 20min. After the supernatant was concentrated, 95% ethanol was added to make the alcohol content reach 65%. spare.

Add 6 times the amount of 70% ethanol to 375 g of Clematis clematis and 300 g of turmeric, reflux twice for 1 hour each time, combine the ethanol extracts, and recover ethanol. And combined with the medicinal solution after water extraction and alcohol precipitation of Shichangpu and peppermint, and concentrated under reduced pressure. Add Tween 80 to solubilize the volatile oil, mix it evenly with the liquid medicine, set the volume to 1000ml, and fill it.

Embodiment 4: the preparation of spray

Clematis 500g, Curcuma 375g, Shichangpu 75g, Peppermint 225g, add 10 times the amount of 70% ethanol to reflux extraction 3 times, each time for 3 hours, the extracts are combined, filtered, the filtrate is added with 0.05% ethyl paraben, 1.5 % stevioside, fixed to 1000ml, distributed in spray bottles, and sterilized.

Embodiment 5: the preparation of buccal tablet

Medicinal materials Clematis 150g, Curcuma 375g, Shichangpu 150g, Peppermint 225g, add 10 times the amount of water, extract the volatile oil by steam distillation, collect the volatile oil in 7 hours, dissolve it with a small amount of ethanol for later use; Add 8 times the amount of 60% ethanol to the residue, reflux twice, 1 hour each time, combine the ethanol extracts, recover the ethanol until it has no alcohol smell, and obtain a concentrated solution, add appropriate amount of mannitol, aspartame, magnesium stearate and volatile oil ethanol solution, mixed evenly, granulated, and compressed into 1000 tablets.

Embodiment 6: Application 1 of Chinese medicine invention composition

Clematis 500g, turmeric 375g, calamus 75g, mint 225g, dextrin 110g, micronized silica gel 15g, made into 1000 capsules, used for the treatment of various acute and chronic coughs.

Embodiment 7: Application 2 of the invention composition of traditional Chinese medicine

Medicinal materials Clematis 300g, Curcuma 250g, Shichangpu 150g, Peppermint 150g, sucrose 90g, stevioside 5g, benzoic acid 10g, make oral liquid 1000ml, for the treatment of infectious cough.

Claims (25)

1. a kind of Chinese medicine composition for treating cough after common cold, is made up of the bulk drug of following weight portion：6-20 gram of the root of Chinese clematis, 6-20 gram of root tuber of aromatic turmeric, 3-10 gram of grass-leaved sweetflag, 3-10 gram of peppermint.

2. Chinese medicine composition according to claim 1, it is characterised in that the Chinese medicine composition by following weight portion bulk drug Composition：10 grams of the root of Chinese clematis, 10 grams of root tuber of aromatic turmeric, 6 grams of grass-leaved sweetflag, 6 grams of peppermint.

3. Chinese medicine composition according to claim 1, it is characterised in that the Chinese medicine composition by following weight portion bulk drug Composition：6 grams of the root of Chinese clematis, 15 grams of root tuber of aromatic turmeric, 6 grams of grass-leaved sweetflag, 9 grams of peppermint.

4. Chinese medicine composition according to claim 1, it is characterised in that the Chinese medicine composition by following weight portion bulk drug Composition：12 grams of the root of Chinese clematis, 10 grams of root tuber of aromatic turmeric, 6 grams of grass-leaved sweetflag, 6 grams of peppermint.

5. Chinese medicine composition according to claim 1, it is characterised in that the Chinese medicine composition by following weight portion bulk drug Composition：15 grams of the root of Chinese clematis, 12 grams of root tuber of aromatic turmeric, 9 grams of grass-leaved sweetflag, 3 grams of peppermint.

6. Chinese medicine composition according to claim 1, it is characterised in that the Chinese medicine composition by following weight portion bulk drug Composition：20 grams of the root of Chinese clematis, 15 grams of root tuber of aromatic turmeric, 3 grams of grass-leaved sweetflag, 9 grams of peppermint.

7. the Chinese medicine composition according to any claim in claim 1 to 6, it is characterised in that the Chinese medicine composition Preparation method be：The root of Chinese clematis, peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, add medicinal material total amount 6-12 times water, with steam distillation Method extracts volatile oil 5-9 hours, collects volatile oil, adds Tween-80 solubilising, standby；Steam distillation extract is filtered, Oily mother liquor and filter residue must be carried；Filter residue adds 6-10 times of decocting to boil 1-3 time, and each 1-3 hours merge decoction liquor and carry oily mother liquor, 75-95% ethanol will be added after supernatant concentration to make alcohol content up to 65-85%, refrigeration is taken out after standing, and is filtered, and filtrate decompression is reclaimed Ethanol is to without alcohol taste；Volatile oil and conventional pharmaceutical adjuvants after solubilising are added, using formulation method conventional in pharmacy of Chinese materia medica, Make suitable dosage forms.

8. Chinese medicine composition according to claim 7, it is characterised in that the preparation method of the Chinese medicine composition is：The root of Chinese clematis, Peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, add medicinal material 12 times of water of total amount, with extraction by steam distillation volatile oil 7 hours, collect Volatile oil, mother liquor is filtered, and adds Tween-80 solubilising, standby；Steam distillation extract is filtered, and must carry oily mother liquor and filter residue： Filter residue adds 10 times of decoctings to boil 3 times, 2 hours every time, merges decoction liquor and carries oily mother liquor, will Jia 95% after supernatant concentration Ethanol makes alcohol content up to 65%, and refrigeration stands and taken out after 15h, filtration, and decompression filtrate recycling ethanol is to without alcohol taste；Add solubilising Volatile oil afterwards and conventional pharmaceutical adjuvants, using formulation method conventional in pharmacy of Chinese materia medica, make granule, oral tablet, Buccal tablet, dispersible tablet, hard capsule, soft capsule, oral liquid, spray, powder-type spray.

9. Chinese medicine composition oral liquid according to claim 8, it is characterised in that the preparation method of the Chinese medicine composition is： The root of Chinese clematis, peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, add medicinal material 12 times of water of total amount, little with extraction by steam distillation volatile oil 7 When, volatile oil is collected, Tween-80 solubilising is added, it is standby；Steam distillation extract is filtered, oily mother liquor and filter residue must be carried； Filter residue adds 10 times of decoctings to boil 3 times, 2 hours every time, merges decoction liquor and carries oily mother liquor, will Jia 95% after supernatant concentration Ethanol makes alcohol content up to 65%, and refrigeration stands and taken out after 15h, filtration, and decompression filtrate recycling ethanol is to without alcohol taste；Add solubilising Volatile oil afterwards and simple syrup, stevioside and Sodium Benzoate, constant volume in being sub-packed in oral liquid bottle, is obtained final product.

10. the Chinese medicine composition according to any claim in claim 1 to 6, it is characterised in that the Chinese medicine composition Preparation method be：The root of Chinese clematis, peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, the 50-80% ethanol for adding medicinal material total amount 5-15 times is returned Stream is extracted, extraction time 1-4 time, each 0.5-4 hours, and alcohol extract merges, and is filtered, filtrate concentration, is added conventional medicinal auxiliary Material, using formulation method conventional in pharmacy of Chinese materia medica, makes suitable dosage forms.

11. Chinese medicine compositions according to claim 10, it is characterised in that the preparation method of the Chinese medicine composition is：Wheeling Celestial being, peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, plus 70% alcohol reflux extract 2 times, 3 hours every time, alcohol extract merge, filter, Concentration, adds conventional pharmaceutical adjuvants, using formulation method conventional in pharmacy of Chinese materia medica, makes granule, oral tablet, mouth containing Piece, dispersible tablet, hard capsule, soft capsule, oral liquid, spray, powder-type spray.

12. Chinese medicine composition oral liquids according to claim 11, it is characterised in that the preparation method of the Chinese medicine composition is： The root of Chinese clematis, peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, plus 70% alcohol reflux extract 2 times, 3 hours every time, alcohol extract merge, Filter, concentration adds simple syrup, stevioside and Sodium Benzoate, constant volume, in being sub-packed in oral liquid bottle, obtains final product.

13. Chinese medicine compositions according to any claim in claim 1 to 6, it is characterised in that the Chinese medicine composition Preparation method be：The root of Chinese clematis, peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, add medicinal material total amount 6-12 times water, with steam distillation Method extracts volatile oil 5-9 hours, collects volatile oil, and beta-cyclodextrin inclusion compound obtains inclusion compound standby；Steam distillation is extracted Liquid is filtered, and must carry oily mother liquor and filter residue；Filter residue add 6-10 times of decocting boil 1-3 time, each 1-3 hours, merge decoction liquor and Oily mother liquor is carried, 75-95% ethanol will be added to make alcohol content up to 65-85% after supernatant concentration, refrigeration is taken out after standing, and is filtered, filter Liquid decompression recycling ethanol then proceedes to be concentrated into thick paste to without alcohol taste, is dried, and gained dried object mixes with volatile oil clathrate compound It is even, conventional pharmaceutical adjuvants are added, using formulation method conventional in pharmacy of Chinese materia medica, make suitable dosage forms.

14. Chinese medicine compositions according to claim 13, it is characterised in that the preparation method of the Chinese medicine composition is：Wheeling Celestial being, peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, add medicinal material 12 times of water of total amount, with extraction by steam distillation volatile oil 7 hours, Volatile oil is collected, beta-cyclodextrin inclusion compound, refrigerated overnight, filtration, low temperature drying obtains Benexate Hydrochloride standby； Steam distillation extract is filtered, oily mother liquor and filter residue must be carried；Filter residue adds 10 times of decoctings to boil 3 times, 2 hours every time, closes And decoction liquor with carry oily mother liquor, will be after supernatant concentration plus 95% ethanol makes alcohol content up to 65%, refrigeration stands and taken out after 15h, Filtration, decompression filtrate recycling ethanol then proceedes to be concentrated into thick paste to without alcohol taste, is dried, gained dried object and inclusion essential oil Thing is well mixed, and adds conventional pharmaceutical adjuvants, using formulation method conventional in pharmacy of Chinese materia medica, make granule, oral tablet, Buccal tablet, dispersible tablet, hard capsule, soft capsule, oral liquid, spray, powder-type spray.

15. Chinese medicine compositions according to claim 14, it is characterised in that the preparation method of the Chinese medicine composition is：Wheeling Celestial being, peppermint, grass-leaved sweetflag and root tuber of aromatic turmeric medicinal material, add medicinal material 12 times of water of total amount, with extraction by steam distillation volatile oil 7 hours, Volatile oil is collected, beta-cyclodextrin inclusion compound, refrigerated overnight, filtration, low temperature drying obtains Benexate Hydrochloride standby； Steam distillation extract is filtered, oily mother liquor and filter residue must be carried；Filter residue adds 10 times of decoctings to boil 3 times, 2 hours every time, closes And decoction liquor with carry oily mother liquor, will be after supernatant concentration plus 95% ethanol makes alcohol content up to 65%, refrigeration stands and taken out after 15h, Filtration, decompression filtrate recycling ethanol then proceedes to be concentrated into thick paste to without alcohol taste, is dried, gained dried object and inclusion essential oil Thing is well mixed, and adds dextrin, cane sugar powder, stevioside, essence, wet granulation, whole grain after being dried to add 0.5% stearic acid Magnesium, is mixed, and is pressed into buccal tablet and obtains final product.

The 16. Chinese medicine composition method of quality control according to any claim in claim 1 to 6, it is characterised in that The discrimination method of the Chinese medicine composition includes one or more in following method：

(1) Chinese medicinal composition preparation 2-10mL for taking the present invention is placed in round bottom sesame seed cake, adds the HCl 30-100mL of 2mol/L, Water-bath backflow 1-3 hours, are placed in separatory funnel after cooling, and add ethyl acetate to extract 1-3 time, and each 15-30mL merges Extract pressurization is evaporated, methanol constant volume to 5-10mL, as need testing solution.Separately take oleanolic acid and the normal aglycon pair of ivy soap Every 1mL is made containing 1mg, according to product plus methyl alcohol as reference substance solution.According to thin-layered chromatography (《Chinese Pharmacopoeia》2010 editions annex VI B) test, each 1-5 μ l of above two solution are drawn, put respectively on same silica gel g thin-layer plate, with petroleum ether-ethyl acetate - formic acid (5-20: 1-10: 0.1-1) is solvent, and lamellae puts presaturation 30 minutes in expansion cylinder, is launched, and is taken out, and is dried, Spray is heated to spot development clear with 10% ethanol solution of sulfuric acid at 105 DEG C.In test sample chromatogram, with reference substance chromatogram phase On the position answered, show the spot of same color.

(2) Chinese medicinal composition preparation 2-10mL for taking the present invention is placed in separatory funnel, addition petroleum ether extraction 1-3 time, every time 5-15mL, merges ether, 2-10mL methanol constant volumes is added after evaporated under reduced pressure, as need testing solution.Separately take peppermint comparison medicine Material, crushed 40 mesh sieves, took 0.5g plus 5-15mL petroleum ethers ultrasound 30min, and after filtration, filtrate is evaporated and is settled to 1mL, As control medicinal material solution.Separately take menthol reference substance plus methyl alcohol makes every 1ml containing 2mg, as reference substance solution.According to thin layer Chromatography (《Chinese Pharmacopoeia》2010 editions annex VI B) test, each 1-5 μ l of above two solution are drawn, put respectively in same silica gel On G lamellaes, with toluene-ethyl acetate (5-20: 1-5) as solvent, after presaturation 30min, launch, take out, dry, With 2% vanillin-sulfuric acid-ethanol (1: 4) developer, 105 DEG C to be heated to spot development clear for spray.In test sample chromatogram, with On reference substance, the corresponding position of control medicinal material chromatogram, show the spot of same color.

(3) Chinese medicinal composition preparation 2-10mL of the present invention is taken, absolute ethyl alcohol 25-50mL ultrasound 15-30min are added, is filtered, Decompression and solvent recovery, residue 1mL ethanol dissolves, used as need testing solution.Root tuber of aromatic turmeric control medicinal material is separately taken, 40 mesh were crushed Sieve, takes 0.5-2g, plus 5-50mL EtOH Sonicate 30min, filters, and is settled to 1mL as control medicinal material solution.According to thin layer color Spectrometry (《Chinese Pharmacopoeia》2010 editions annex VI B) test, each 1-5 μ l of above two solution are drawn, put respectively in same silica G On lamellae, with petroleum ether (60-90 DEG C)-ethyl acetate (10-30: 5-10) as solvent, after presaturation 30min, exhibition Open, taking-up is dried, spray with 10% concentrated sulfuric acid ethanol developer, 105 DEG C to be heated to spot development clear.In test sample chromatogram, On position corresponding with control medicinal material chromatogram, show the spot of same color.

(4) Chinese medicinal composition preparation 2-10mL for taking the present invention is placed in separatory funnel, addition petroleum ether extraction 1-3 time, every time 5-15mL, merges ether, 2-10mL methanol constant volumes is added after evaporated under reduced pressure, as need testing solution.Separately take grass-leaved sweetflag control Medicinal material, crushed 40 mesh sieves, took 0.5-2g, plus 10-40mL ether ultrasound 30min, and after filtration, filtrate is evaporated and is settled to 1mL, as need testing solution.Separately take α-asarone reference substance plus methyl alcohol is prepared into every mL containing 2mg, as reference substance solution. According to thin-layered chromatography (《Chinese Pharmacopoeia》2010 editions annex VI B) test, each 1-5 μ l of above two solution are drawn, put in same respectively On one silica gel g thin-layer plate, with petroleum ether (60-90 DEG C)-ethyl acetate (5-20: 2-20) as solvent, presaturation 30min Afterwards, launch, take out, dry, put and inspected under ultraviolet lamp (365nm).In test sample chromatogram, with reference substance solution chromatogram phase On the position answered, show the spot of same color.

17. Chinese medicine composition method of quality control according to claim 16, it is characterised in that the discriminating of the Chinese medicine composition Method includes one or more in following method：

(1) Chinese medicinal composition preparation 2mL for taking the present invention is placed in round bottom sesame seed cake, adds the HCl 30mL of 2mol/L, water-bath Backflow 2 hours, is placed in separatory funnel after cooling, and adds ethyl acetate to extract 3 times, each 15mL, combining extraction liquid pressurization It is evaporated, methanol constant volume to 10mL, as need testing solution.Separately take oleanolic acid and the normal aglycon reference substance of ivy soap plus methyl alcohol system Contain 1mg into every 1mL, as reference substance solution.According to thin-layered chromatography (《Chinese Pharmacopoeia》2010 editions annex VI B) test, inhale The each 5 μ l of above two solution are taken, is put respectively on same silica gel g thin-layer plate, with petroleum ether-ethyl acetate-formic acid (10: 3: 0.1) For solvent, lamellae puts presaturation 30 minutes in expansion cylinder, launches, and takes out, and dries, and sprays with 10% ethanol solution of sulfuric acid, It is clear spot development to be heated at 105 DEG C.In test sample chromatogram, on position corresponding with reference substance chromatogram, show same color Spot.

(2) Chinese medicinal composition preparation 2mL for taking the present invention is placed in separatory funnel, addition petroleum ether extraction 3 times, each 5mL, Merge ether, 2mL methanol constant volumes are added after evaporated under reduced pressure, as need testing solution.Peppermint control medicinal material is separately taken, 40 were crushed Mesh sieve, takes 0.5g plus 10mL petroleum ethers ultrasound 30min, and after filtration, filtrate is evaporated and is settled to 1mL, used as control medicinal material Solution.Separately take menthol reference substance plus methyl alcohol makes every 1ml containing 2mg, as reference substance solution.According to thin-layered chromatography (《China Pharmacopeia》2010 editions annex VI B) test, each 2 μ l of above two solution are drawn, put respectively on same silica gel g thin-layer plate, with Toluene-ethyl acetate (8: 18) is solvent, after presaturation 30mmin, is launched, and is taken out, and is dried, and is sprayed with 2% vanillic aldehyde sulphur Acid-ethanol (1: 4) developer, 105 DEG C to be heated to spot development clear.In test sample chromatogram, with reference substance, control medicinal material On the corresponding position of chromatogram, show the spot of same color.

(3) Chinese medicinal composition preparation 2mL of the present invention is taken, adds absolute ethyl alcohol 25mL ultrasound 15min, filtration to reduce pressure back Solvent is received, residue 1mL ethanol dissolves, used as need testing solution.Root tuber of aromatic turmeric control medicinal material is separately taken, 40 mesh sieves were crushed, is taken 2g, plus 25mL EtOH Sonicate 30min, filter, and are settled to 1mL as control medicinal material solution.According to thin-layered chromatography (《China Pharmacopeia》2010 editions annex VI B) test, each 1-5 μ l of above two solution are drawn, put respectively on same silica gel g thin-layer plate, With petroleum ether (60-90 DEG C)-Ethyl formate (15: 5) as solvent, after presaturation 30min, launch, taking-up is dried, and is sprayed With 10% concentrated sulfuric acid ethanol developer, 105 DEG C to be heated to spot development clear.In test sample chromatogram, with control medicinal material chromatogram phase On the position answered, show the spot of same color.

(4) Chinese medicinal composition preparation 5mL for taking the present invention is placed in separatory funnel, addition petroleum ether extraction 3 times, each 5mL, Merge ether, 5mL methanol constant volumes are added after evaporated under reduced pressure, as need testing solution.Grass-leaved sweetflag control medicinal material is separately taken, was crushed 40 mesh sieves, take 1g, plus 10mL ether ultrasound 30min, and after filtration, filtrate is evaporated and is settled to 1mL, molten as test sample Liquid.Separately take α-asarone reference substance plus methyl alcohol is prepared into every mL containing 2mg, as reference substance solution.According to thin-layered chromatography (《In State's pharmacopeia》2010 editions annex VI B) test, each 1-5 μ l of above two solution are drawn, put respectively on same silica gel g thin-layer plate, With petroleum ether (60-90 DEG C)-ethyl acetate (6: 4) as solvent, after presaturation 30min, launch, take out, dry, Put and inspected under ultraviolet lamp (365nm).In test sample chromatogram, on position corresponding with reference substance solution chromatogram, show same color Spot.

The 18. Chinese medicine composition method of quality control according to any claim in claim 1 to 6, it is characterised in that The assay of the Chinese medicine composition include following assay method:

Chromatographic condition：Octadecylsilane chemically bonded silica is filler；Mobile phase is acetonitrile (A) -0.1-0.5% phosphoric acid (B), Gradient elution (0～15min：70%A, 15～20min：70% → 90%A, 20～30min：90%A, 30～35：90% → 70%)； 30 DEG C of column temperature；Flow velocity 1.0mlmin-1, Detection wavelength 205nm.External standard method peak area quantification.

It is prepared by reference substance solution：Hederagenin, the oleanolic acid reference substance in 60 DEG C of drying under reduced pressure 3-5 hours is weighed, plus Methyl alcohol is made concentration of every 1ml containing 20ug and is obtained final product.

It is prepared by need testing solution：Chinese medicinal composition preparation content 1-5ml, plus 2mol/L hydrochloric acid solution 10-50ml are measured, plus Heat backflow 1-3 hours, cool down immediately, and in moving into separatory funnel, plus ethyl acetate shaking is extracted 1-3 time, each 10-30ml, Combined ethyl acetate liquid, is evaporated, plus methyl alcohol dissolving, and in being transferred to 5ml measuring bottles, constant volume is obtained final product.

Determination method：Reference substance solution and each 10ul of need testing solution are drawn respectively, liquid chromatograph is injected, is determined, obtain final product.

Per milliliter gram must not be less than containing the root of Chinese clematis in terms of hederagenin in the oral liquid of Chinese medicine composition of the present invention 0.60-1.00mg, must not be less than 0.10-0.40mg in terms of oleanolic acid.

19. content assaying methods according to claim 18, it is characterised in that the method includes following assay method：

Chromatographic condition：Octadecylsilane chemically bonded silica is filler；Mobile phase be the phosphoric acid (B) of acetonitrile (A) -0.1%, gradient Wash-out (0～15min：70%A, 15～20min：70% → 90%A, 20～30min：90%A, 30～35：90% → 70%)；Post 30 DEG C of temperature；Flow velocity 1.0mlmin-1, Detection wavelength 205nm.External standard method peak area quantification.

It is prepared by reference substance solution：Weigh in 60 DEG C of drying under reduced pressure hederagenin of 4 hours, oleanolic acid reference substance, plus first Alcohol is made concentration of every 1m1 containing 20ug and is obtained final product.

It is prepared by need testing solution：Chinese medicinal composition preparation content 1ml, plus 2mol/L hydrochloric acid solution 30ml are measured, is heated back Stream 3 hours, cools down immediately, and in moving into separatory funnel, plus ethyl acetate shaking is extracted 3 times, and each 15ml merges acetic acid second Ester liquid, is evaporated, plus methyl alcohol dissolving, and in being transferred to 5ml measuring bottles, constant volume is obtained final product.

Determination method：Reference substance solution and each 10ul of need testing solution are drawn respectively, liquid chromatograph is injected, is determined, obtain final product.

Per milliliter gram must not be less than 0.70mg containing the root of Chinese clematis in terms of hederagenin in the oral liquid of Chinese medicine composition of the present invention, with Oleanolic acid meter must not be less than 0.25mg.

20. content assaying methods as described in claim 18 or 19, it is characterised in that wherein methyl alcohol can with 70% ethanol or Ethyl acetate is substituted.

21. content assaying methods as described in claim 18 or 19, it is characterised in that wherein ultrasonic processing method can be used back Stream extracting method is substituted.

22. content assaying methods as described in claim 18 or 19, it is characterised in that wherein mobile phase ultrasonic processing method can To be substituted with reflux extraction method.

23. content assaying methods as described in claim 18 or 19, it is characterised in that the wherein phosphoric acid of mobile phase acetonitrile -0.1% Can be substituted with acetonitrile-water.

24. Chinese medicine compositions according to any claim in claim 1 to 6, it is characterised in that the Chinese medicine composition exists Prepare the application in the infectious cough medicine for the treatment of.

25. Chinese medicine compositions according to any claim in claim 1 to 6, it is characterised in that the Chinese medicine composition Application in all kinds of chronic cough medicines for the treatment of are prepared.