CN1754554A - Traditional Chinese medicine compositions - Google Patents
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Abstract
The invention discloses a medicinal composition for inhibiting influenza virus, upper respiratory tract infection virus, SARS virus and bacteria. The composition comprises earthworm, baikal skullcap root glycoside and licorice root, and can be made into various pharmaceutically acceptable dosage types including tablets, capsule, granules, dripping pills, oral liquids and sprays.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition, particularly be used to suppress the Chinese medicine composition of influenza virus, upper respiratory tract infection virus, SARS virus and antibacterial.
Background technology
Influenza claims influenza again, it is a kind of acute respiratory infectious disease that causes by influenza virus, the influenza morbidity is not subject to seasonal restrictions, all can take place throughout the year, but with autumn and winter season sickness rate the highest, because influenza virus is present in the secretions such as patient's nasal mucus, mouthful saliva, sputum, very easily infects and cause popular.According to the statistics of World Health Organization (WHO), the annual world has 600,000,000 to 1,200,000,000 people to infect influenza, dead 250,000 to 500,000 people, and the sickness rate of influenza is 15.6%.So far still needleless to the desirable Western medicine of virus.Influenza belongs to the category of pestilence in the traditional Chinese medical science, and the Chinese medicine prevention influenza has certain advantage, but does not also act on clear and definite Chinese patent medicine.
Upper respiratory tract infection is by due to antibacterial and the virus, and particularly viral upper respiratory tract infection is one of human modal disease, and its symptom can gently can weigh, and at present medical circle does not still have specific drug, in the treatment only to support symptomatic treatment.Upper respiratory tract infection belongs to the exterior syndrome category in the traditional Chinese medical science, its disease be found in " TAI YANG syndrome ", " Shaoyang disease " in " pathogenic wind-warm ", " spring-warm syndrome " in the pestilence, " fever disease in summer " etc. and the typhoid fever, " YANG MING syndrome " various in.Chinese medicine has certain advantage in the anti-disease of effecting a permanent cure than doctor trained in Western medicine.Though at present the domestic Chinese patent medicine that has had certain this disease of treatment also exists pathogenesis not have unified understanding, particularly at virus, the new drug that has killing and suppress virus function extremely lacks.
Severe acute respiratory syndrome (Infectious Atypical Pneumonia, IAP) WHO is called severe acute respiratory syndrome (Severe Acute Respiratory Syndroma again, SARS), be the extremely strong respiratory system disease of a kind of infectiousness, belong to " science of epidemic febrile disease of Chinese medicine " category of the traditional Chinese medical science.The primary disease winter-spring season that starts is characterized in morbidity sharply, and the change of disease is rapid, is dangerously ill, and infectiousness is extremely strong, the general susceptible of crowd and cause popular.Popular in more than 30 countries of the China and the world, SARS epidemic period in 2003, case 8422 examples take place in the whole world altogether, dead 916 examples, China's Mainland 5327 examples wherein, dead 349 examples, the serious harm people's life and health, and social stability and economic construction brought negative effect.
Pathogenesis at the SARS traditional Chinese medical science lapses to, and implements determination of treatment based on pathogenesis obtained through differentiation of symptoms and signs, and SARS invades lung for the epidemic disease poison in early days, the wet thermal resistance of holding back, suitable heat-clearing and toxic substances removing, removing dampness expelling pathogenic factors from the exterior; Mid-term, pulmonary's excess pathogen filled stagnantly, and pyretic toxicity, stasis of blood poison, noxious dampness are stopped up and held back the lung network, and the mechanism of qi obturation is because of the real void that causes, suitable benefiting qi and removing blood stasis, eliminating damp, detoxifying; The later stage asthenia of pulmonosplenic qi, painstaking effort consumes focuses on invigorating the spleen and regulating the stomach, spleen rises stomach and falls, in be so incensed that to answer, painstaking effort are spontaneous.Prescription mostly is decoction, and recommending with Chinese patent medicine is QINKAILING ZHUSHEYE, Herba Erigerontis injection, HUOXIANG ZHENGQI YE, berberine, SHUANGHUANGLIAN KOUFUYE etc.Do not have at present in the Chinese patent medicine at the special-purpose medicine of the control of SARS virus, and not at the use of SARS virus control, the dosage form of carrying convenience.
Western medical treatment at SARS, at the treatment of heating, mainly be antipyretic analgesic, in early days as aspirin, ibuprofen, diclofenac, indomethacin etc., but in various degree gastrointestinal reaction is arranged more, or anaphylaxis arranged, general medication 3 days, body temperature are not fallen and should not be continued to use, and life-time service also can suppress hemopoietic function of bone marrow.Cause leukocyte and hematoblastic minimizing.Treatment for SARS hyperpyrexia and respiratory distress syndrome, doctor trained in Western medicine is generally used Physical temperature-lowering, antibiotic and hormone therapy, the particularly use of hormone though can temporarily stablize the state of an illness, also may be covered some symptoms, after the drug withdrawal, body temperature can go up rapidly, simultaneously because the infection deficiency causes the rapid diffusion of infection focus.
In view of the difficulty of above-mentioned harm humans life and health diseases prevention and treatment, this research is to develop has the compound Chinese medicinal preparation that suppresses influenza virus, upper respiratory tract infection virus, SARS virus and antibacterial.
Summary of the invention
One object of the present invention is to disclose a kind of new Chinese medicine composition that is used to suppress influenza virus, upper respiratory tract infection virus, SARS virus and antibacterial; Another object of the present invention is to disclose a kind of new Chinese medicine composition composition spray that is used to suppress influenza virus, upper respiratory tract infection virus, SARS virus and antibacterial; Another object of the present invention is a kind of new Chinese medicine composition method that is used to suppress influenza virus, upper respiratory tract infection virus, SARS virus and antibacterial of open preparation; Another object of the present invention is to disclose a kind of new being used to and prepares the medicinal herb spray method of quality control that suppresses influenza virus, upper respiratory tract infection virus, SARS virus and antibacterial.
The crude drug of pharmaceutical composition of the present invention is formed and proportioning following (by weight): Pheretima 100-500 weight portion baicalin 1-10 weight portion Radix Glycyrrhizae 100-500 weight portion.
The crude drug of pharmaceutical composition of the present invention is formed and proportion optimization following (by weight): Pheretima 200-400 weight portion baicalin 3-8 weight portion Radix Glycyrrhizae 200-400 weight portion.
The crude drug of pharmaceutical composition of the present invention is formed and proportion optimization following (by weight): Pheretima 300 weight portion baicalins 6.5 weight portion Radix Glycyrrhizaes 300 weight portions.
The preparation method of Chinese medicine composition of the present invention is:
Get Pheretima after the pretreatment, acetic acid-the sodium-acetate buffer that adds 0.1mol/L, mixing was placed after 15-25 hour under the room temperature, 15-25 ℃ of 400-600rpm homogenate 10-20 minute, 8000-12000rpm is centrifugal, gets supernatant and puts placement in 50-65 ℃, and 8000-12000rpm is centrifugal, get supernatant and carry out ultrafiltration dialysis, concentrated solution cold preservation is standby; Radix Glycyrrhizae decocts with water 1-3 time, each 1-3 hour, collecting decoction filtered, filtrate is concentrated into relative density 1.10, add ethanol and make and contain the alcohol amount and reach 60-70%, left standstill 10-14 hour, filter, reclaim ethanol, add the Pheretima extracting solution, stir evenly, suitable with the ammonia adjust pH, add baicalin, it is an amount of to add water, stirs evenly cold preservation, spend the night, filter, directly or indirectly add pharmaceutically acceptable excipient through conventional operation and make clinical acceptable forms, as tablet, capsule, granule, drop pill, oral liquid, spray.
Chinese medicine composition spray preparing process of the present invention is:
Pheretima 100-500 weight portion baicalin 1-10 weight portion Radix Glycyrrhizae 100-500 weight portion.
Get Pheretima after the pretreatment, acetic acid-the sodium-acetate buffer that adds 0.1mol/L, mixing was placed after 15-25 hour under the room temperature, 15-25 ℃ of 400-600rpm homogenate 10-20 minute, 8000-12000rpm is centrifugal, gets supernatant and puts placement in 50-65 ℃, and 8000-12000rpm is centrifugal, get supernatant and carry out ultrafiltration dialysis, concentrated solution cold preservation is standby; Radix Glycyrrhizae decocts with water 1-3 time, and each 1-3 hour, collecting decoction filtered, filtrate is concentrated into relative density 1.10, adds ethanol and makes and contain the alcohol amount and reach 60-70%, leaves standstill 10-14 hour, filters, reclaim ethanol, add the Pheretima extracting solution, stir evenly, suitable with the ammonia adjust pH, add baicalin, it is an amount of to add water, stirs evenly cold preservation, spend the night, filter, add 0.2% sodium benzoate and 0.6% apple essence, stir evenly, filter, add water and make 1000 parts by volume, fill, promptly.Weight portion/parts by volume described in the description of the present invention is corresponding with grams per milliliter.
The method of quality control of Chinese medicine composition composition spray of the present invention comprises following discriminating and/or assay: discrimination method comprises a kind of and/or several in the following method:
1. Radix Scutellariae: get this product liquid 10ml, water bath method, residue add methanol 5ml thermosol, are concentrated into 1ml, as need testing solution; Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (appendix VIB), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, with 4-6: 1-4: 1-2: 1-2 ethyl acetate-butanone-formic acid-water developing solvent, pre-equilibration 25-35 minute, launch, take out, dry, spray is with 1% ferric chloride alcoholic solution; In the test sample chromatograph with baicalin reference substance chromatograph relevant position on show an identical dirty-green speckle;
2. Radix Glycyrrhizae: get this product liquid 10ml, add methanol 50ml reflux, extract, 1 hour, reclaim methanol, add 30ml water and make dissolving to doing, with n-butanol extraction 2-4 time, each 20ml, merging n-butyl alcohol liquid, evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g in addition, with the test sample method operate solution 0.5ml, medical material solution in contrast.Test according to thin layer chromatography (appendix VIB), draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same prefabricated silica gel g thin-layer plate with the 1%NaOH solution spray, with 13-17: 1-2: 1-2: 1-3 ethyl acetate-formic acid-glacial acetic acid-water developing solvent, launch, take out, dry, spray is put under the ultra-violet lamp and is inspected with 10% ethanol solution of sulfuric acid; In the test sample chromatograph with control medicinal material chromatograph relevant position on show the fluorescence speckle of a same color;
3. Pheretima: get this product 10ml, add chloroform 20ml, close plug, supersound process 15-25 minute, filter, filtrate is concentrated into about 1ml, as need testing solution.Other gets Pheretima medical material 3g, makes medical material solution with method; According to thin layer chromatography (appendix VIB) test, draw each 5 μ l of above-mentioned two clock solution, put respectively on same silica gel g thin-layer plate, with 7-11: 1-2 benzene one ethyl acetate is developing solvent, launches, and takes out, and dries, and puts under the ultra-violet lamp and inspects; In the test sample chromatograph, with the corresponding position of medical material chromatograph on, show the fluorescence speckle of same color.
Assay:
Baicalin: measure according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000)
Chromatographic condition and system suitability test: with octadecylsilane chemically bonded silica is filler; 45-50: 50-55: the 0.1-0.4 methanol-water-phosphoric acid is a mobile phase; The detection wavelength is 280nm; Number of theoretical plate calculates by the baicalin peak should be not less than 2500;
The preparation of reference substance solution: precision takes by weighing at 4 hours baicalin reference substance of 60 ℃ of drying under reduced pressure an amount of, adds methanol and makes the solution that every 1ml contains 64.40 μ g;
The preparation of need testing solution: precision is measured this product solution 1ml and is put in the 100ml measuring bottle, adds methanol to scale, shakes up, and puts slight fever in the water-bath, promptly;
Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject hplc determination, external standard method is calculated, promptly; The every 25ml liquid of this product contains baicalin (C
21H
18O
11) must not be less than 67.5mg.
Present composition preparation is mainly used in the control of influenza, upper respiratory tract infection and SARS, effect with good heat-clearing and toxic substances removing, blood circulation promoting and blood stasis dispelling, expelling phlegm for arresting cough reaches and prevents and treats influenza, upper respiratory tract infection and SARS purpose in order to improve respiratory tract and pulmonary function by antibiotic, antiviral, antiinflammatory, antiallergic, immunomodulating and work such as analgesic.
The made liquid spray dosage form of the present composition has that onset is rapid, and toxic and side effects is little, accumulating, carry, characteristics easy to use.The medicine finished product is the water soluble preparation of effective ingredient, satisfy pharmacodynamics to the preparation requirement in, have easy to prepare, the industrialization advantage that waits with low cost.
The method of quality control of present composition spray makes the spray steady quality, is easy to control.Following experimental example further specifies the present invention.
Experimental example 1 present composition composition screening experiment
After the equilibrium of 160 male mice body weight, random group, press orthogonal array L16 (215) intension, five factors, two levels that adapt to this research are observed requirement, and five kinds of influence factors are: A Pheretima, B baicalin glycoside, C glycyrrhizic acid, D naloxone, E gardenoside and Radix Notoginseng total glucosides.Each factor all has two states to be: not medication (representing with 1) and medication (representing with 2).Shown in the specific design statistics form, 15 row on project hurdle are represented 5 kinds of factors or its reciprocal action in twos, wherein the 01st, 02,04,08,15 row are represented 5 factor affecting effects such as A, B, C, D, E respectively, the reciprocal action in twos between all the other each each positions of row reflection; 16 row of laboratory animal are represented 16 treated animals; Ri represents the result of observation index, carries out the t check between 1 level, each index value of 2 levels, analyzes the difference of two levels, with the have statistical significance of P＜0.25 for effective trend
1. single factors is to the influence of pulmonary edema degree
1. Pheretima
80 lung weight in wet bases of not using the Pheretima mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.1048 ± 0.037286,0.022613 ± 0.008968,0.082188 ± 0.032733,0.774642 ± 0.088514,80 lung weight in wet bases of using the Pheretima mice, the lung dry weight, water is heavy, the mean values of water percentage ratio increases to 0.116613 ± 0.03751 respectively, 0.024288 ± 0.008728,0.092325 ± 0.031154,0.78954 ± 0.05269, the lung weight in wet base, water is heavy, the increase of water percentage ratio has significance meaning (P＜0.05), and the lung dry weight reduces no significance meaning (P＞0.05).Illustrate that Pheretima may have negative effects to chmice acute respiratory distress syndrome pulmonary edema.
2. baicalin
80 lung weight in wet bases of not using the baicalin mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.103638 ± 0.036055,0.022263 ± 0.009157,0.081375 ± 0.030975,0.77920 ± 0.088845,80 lung weight in wet bases of using the baicalin mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.117775 ± 0.038297 respectively, 0.024638 ± 0.008445,0.093138 ± 0.032627,0.784974 ± 0.053037, the lung weight in wet base, the lung dry weight, water heavily increases has the significance meaning (P＜0.05), and water percentage ratio reduces no significance meaning (P＞0.05).Illustrate that baicalin may have negative effects to chmice acute respiratory distress syndrome pulmonary edema.
3. Radix Glycyrrhizae
80 lung weight in wet bases of not using the glycyrrhizic acid mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.10865 ± 0.038944,0.021925 ± 0.008406,0.086725 ± 0.032603,0.795901 ± 0.053888,80 lung weight in wet bases of using the glycyrrhizic acid mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.112763 ± 0.036642 respectively, 0.024975 ± 0.009092,0.087788 ± 0.032102,0.768281 ± 0.086216, the lung dry weight, water percentage ratio reduces significance meaning (P＜0.05), and the lung weight in wet base, water heavily reduces no significance meaning (P＞0.05).Illustrate that glycyrrhizic acid has preventive and therapeutic effect to chmice acute respiratory distress syndrome pulmonary edema.
4. naloxone
80 lung weight in wet bases of not using the naloxone mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.108275 ± 0.039162,0.023088 ± 0.009192,0.085188 ± 0.033605,0.778869 ± 0.082838,80 lung weight in wet bases of using the naloxone mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.113138 ± 0.036362 respectively, 0.023813 ± 0.00856,0.089325 ± 0.03092,0.785313 ± 0.061969, the lung weight in wet base, the lung dry weight, water is heavy, meaning (P＞0.05) that water percentage ratio changes that there are no significant.Illustrate that naloxone does not have preventive and therapeutic effect to chmice acute respiratory distress syndrome pulmonary edema.
5. Radix Notoginseng Fructus Gardeniae (collateral dredging is rescued injection of brain liquid)
80 lung weight in wet bases of not using Radix Notoginseng Fructus Gardeniae mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.114975 ± 0.04063,0.023563 ± 0.009487,0.091413 ± 0.034418,0.791414 ± 0.061221,80 lung weight in wet bases of using Radix Notoginseng Fructus Gardeniae mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.106438 ± 0.034354 respectively, 0.023338 ± 0.008246,0.0831 ± 0.029572,0.772768 ± 0.082458, the lung weight in wet base, the lung dry weight, water is heavy, meaning (P＞0.05) that water percentage ratio changes that there are no significant.Illustrate that the Radix Notoginseng Fructus Gardeniae does not have preventive and therapeutic effect to chmice acute respiratory distress syndrome pulmonary edema.
2. alleviate the dual factors reciprocal action of pulmonary edema degree
1. the reciprocal action of Pheretima and Radix Glycyrrhizae
The lung weight in wet base of 80 Pheretimas and glycyrrhizic acid non-interaction action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.111463 ± 0.038428,0.022813 ± 0.009455,0.08865 ± 0.03106,0.796114 ± 0.058715,80 Pheretimas and glycyrrhizic acid have the lung weight in wet base of reciprocal action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.10995 ± 0.037283 respectively, 0.024088 ± 0.008234,0.085863 ± 0.033547,0.768068 ± 0.082931, water percentage ratio reduces significance meaning (P＜0.01), and the lung weight in wet base, the lung dry weight, water heavily reduces no significance meaning (P＞0.05).Illustrate that Pheretima and glycyrrhizic acid reciprocal action have preventive and therapeutic effect to chmice acute respiratory distress syndrome pulmonary edema.
2. the reciprocal action of baicalin and Radix Glycyrrhizae
The lung weight in wet base of 80 baicalins and Radix Glycyrrhizae non-interaction action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.112625 ± 0.037405,0.022838 ± 0.009678,0.089788 ± 0.030921,0.796264 ± 0.062072, the lung weight in wet base of 80 baicalins and Radix Glycyrrhizae reciprocal action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.108788 ± 0.038226 respectively, 0.024063 ± 0.007975,0.084725 ± 0.033541,0.767918 ± 0.080396, water percentage ratio reduces significance meaning (P＜0.01), and the lung weight in wet base, the lung dry weight, water heavily reduces no significance meaning (P＞0.05).Illustrate that baicalin and Radix Glycyrrhizae reciprocal action have preventive and therapeutic effect to chmice acute respiratory distress syndrome pulmonary edema.
3. the reciprocal action of naloxone and Radix Notoginseng Fructus Gardeniae
The lung weight in wet base of 80 naloxones and Radix Notoginseng Fructus Gardeniae non-interaction action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.107288 ± 0.041068,0.02165 ± 0.00948,0.085638 ± 0.034237,0.795372 ± 0.062714, the lung weight in wet base of 80 naloxones and Radix Notoginseng Fructus Gardeniae reciprocal action mice, dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.114125 ± 0.034024 respectively, 0.02525 ± 0.007847,0.088875 ± 0.030275,0.76881 ± 0.080206, the lung dry weight, water percentage ratio reduces significance meaning (P＜0.01), and the lung weight in wet base, water heavily reduces no significance meaning (P＞0.05).Illustrate that naloxone and Radix Notoginseng Fructus Gardeniae reciprocal action have preventive and therapeutic effect to chmice acute respiratory distress syndrome pulmonary edema.
4. the reciprocal action of Radix Glycyrrhizae and naloxone
The lung weight in wet base of 80 Radix Glycyrrhizaes and naloxone non-interaction action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.1088 ± 0.0371,0.0225 ± 0.0096,0.0863 ± 0.0303,0.7929 ± 0.0639, the lung weight in wet base of 80 Radix Glycyrrhizaes and naloxone reciprocal action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.1126 ± 0.0385 respectively, 0.0244 ± 0.0080,0.0882 ± 0.0342,0.7713 ± 0.0800, water percentage ratio reduces significance meaning (P＜0.05), and the lung weight in wet base, the lung dry weight, water heavily reduces no significance meaning (P＞0.05).Illustrate that Radix Glycyrrhizae and naloxone reciprocal action have preventive and therapeutic effect to chmice acute respiratory distress syndrome pulmonary edema.
3. do not influence the dual factors reciprocal action of pulmonary edema degree
1. the reciprocal action of Pheretima and baicalin
The lung weight in wet base of 80 Pheretimas and baicalin non-interaction action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.104075 ± 0.039398,0.023300 ± 0.009979,0.080775 ± 0.032726,0.770971 ± 0.087163,80 lung weight in wet bases of using the Pheretima mice, the lung dry weight, water is heavy, the mean values of water percentage ratio increases to 0.117338 ± 0.035023 respectively, 0.0236 ± 0.007642,0.093738 ± 0.030627,0.793211 ± 0.053624, the lung weight in wet base, water is heavy, the increase of water percentage ratio has significance meaning (P＜0.05), and the lung dry weight increases no significance meaning (P＞0.05).Illustrate that Pheretima and baicalin reciprocal action have negative effects to chmice acute respiratory distress syndrome pulmonary edema.
2. the reciprocal action of Pheretima and naloxone
The lung weight in wet base of 80 Pheretimas and naloxone non-interaction action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.106438 ± 0.037715,0.02245 ± 0.00911,0.083988 ± 0.032451,0.781732 ± 0.084837, the lung weight in wet base of 80 Pheretimas and naloxone reciprocal action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio increases to 0.114975 ± 0.03753 respectively, 0.02445 ± 0.008545,0.090525 ± 0.031927,0.78245 ± 0.059378, the lung weight in wet base, the lung dry weight, water is heavy, meaning (P＞0.05) that water percentage ratio changes that there are no significant.Illustrate that Pheretima and naloxone reciprocal action do not have preventive and therapeutic effect to chmice acute respiratory distress syndrome pulmonary edema
3. the reciprocal action of baicalin glycoside and naloxone
The lung weight in wet base of the reciprocal action mice of 80 baicalin glycosides and naloxone, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.106625 ± 0.037923,0.02315 ± 0.008828,0.083475 ± 0.032396,0.775055 ± 0.081643, the lung weight in wet base of the reciprocal action mice of 80 baicalin glycosides and naloxone, the lung dry weight, water is heavy, the mean values of water percentage ratio increases to 0.114788 ± 0.037363 respectively, 0.02375 ± 0.008939,0.091038 ± 0.031869,0.789127 ± 0.062908, the lung weight in wet base, water heavily increases has the significance meaning (P＜0.05), and the lung dry weight, water percentage ratio increases no significance meaning (P＞0.05).Illustrate that baicalin glycoside and naloxone reciprocal action are swollen with negative effects to chmice acute respiratory distress syndrome edema caused by the lung disorder.
4. the reciprocal action of Radix Glycyrrhizae and Radix Notoginseng Fructus Gardeniae
Lung weight in wet base, lung dry weight, the water mean values heavy, water percentage ratio of 80 Radix Glycyrrhizaes and Radix Notoginseng Fructus Gardeniae non-interaction action mice is respectively 0.1068 ± 0.0393,0.0226 ± 0.0089,0.0842 ± 0.0339,0.7802 ± 0.0845, lung weight in wet base, lung dry weight, the water mean values heavy, water percentage ratio of 80 Radix Glycyrrhizaes and Radix Notoginseng Fructus Gardeniae reciprocal action mice increases to 0.1146 ± 0.0359,0.0243 ± 0.0088,0.0904 ± 0.0305,0.7840 ± 0.0598 respectively, and lung weight in wet base, lung dry weight, water are heavy, the water percentage ratio meaning (P＞0.05) that increases that there are no significant.Illustrate that Radix Glycyrrhizae and Radix Notoginseng Fructus Gardeniae reciprocal action do not have influence to chmice acute respiratory distress syndrome pulmonary edema.
5. the reciprocal action of baicalin and Radix Notoginseng Fructus Gardeniae
The lung weight in wet base of 80 baicalins and Radix Notoginseng Fructus Gardeniae non-interaction action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.111275 ± 0.036446,0.023625 ± 0.008638,0.08765 ± 0.03059,0.784136 ± 0.058003, the lung weight in wet base of 80 baicalins and Radix Notoginseng Fructus Gardeniae reciprocal action mice, dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.110138 ± 0.039229 respectively, 0.023275 ± 0.009129,0.086863 ± 0.034029,0.780046 ± 0.085735, the lung dry weight, the lung weight in wet base, water is heavy, meaning (P＞0.05) that water percentage ratio reduces that there are no significant.Illustrate that baicalin and Radix Notoginseng Fructus Gardeniae reciprocal action do not have influence to chmice acute respiratory distress syndrome pulmonary edema tool.
6. the reciprocal action of Pheretima and Radix Notoginseng Fructus Gardeniae
The lung weight in wet base of 80 Pheretimas and Radix Notoginseng Fructus Gardeniae non-interaction action mice, the lung dry weight, water is heavy, the mean values of water percentage ratio is respectively 0.107488 ± 0.034436,0.0223 ± 0.008912,0.085188 ± 0.029367,0.789401 ± 0.066445, the lung weight in wet base of 80 Pheretimas and Radix Notoginseng Fructus Gardeniae reciprocal action mice, dry weight, water is heavy, the mean values of water percentage ratio is reduced to 0.113925 ± 0.040756 respectively, 0.024600 ± 0.008713,0.089325 ± 0.034971,0.774781 ± 0.078741, the lung dry weight, the lung weight in wet base, water is heavy, meaning (P＞0.05) that water percentage ratio reduces that there are no significant.Illustrate that Pheretima and Radix Notoginseng Fructus Gardeniae reciprocal action do not have influence to chmice acute respiratory distress syndrome pulmonary edema tool.
4. experiment conclusion
1. single factors reduces the degree of pulmonary edema: Pheretima, baicalin have negative effects, and Radix Glycyrrhizae has preventive and therapeutic effect, and naloxone, Radix Notoginseng Fructus Gardeniae do not have preventive and therapeutic effect.
2. factor alleviates the degree of pulmonary edema alternately: Pheretima and Radix Glycyrrhizae, baicalin and Radix Glycyrrhizae, naloxone and 4 kind of two factor reciprocal action such as Radix Notoginseng Fructus Gardeniae, Radix Glycyrrhizae and naloxone have the effect of control.
In sum, Pheretima, baicalin and Radix Glycyrrhizae prescription therapeutic SARS are best screening compatibility.
The experiment of experimental example 2 Study on extraction
One, extraction process of Glycyrrhiza uralensis research
With reference to " 2000 editions Radix Glycyrrhizae extractum preparation methoies of Chinese pharmacopoeia, it is solvent that Radix Glycyrrhizae is extracted with water, boils number of times, adds the water multiple and the time is factor A with decocting, and it is factor B that decocting liquid concentrates relative density, ethanol precipitation concentration is factor C, adopts four factors, three water-glass experiment arrangement; The detection index is a glycyrrhizic acid content; Detection method adopts the HPLC method.
Radix Glycyrrhizae content assaying method: with reference to content assaying method under pharmacopeia (Chinese Pharmacopoeia 2000 version one one 66 pages) the Radix Glycyrrhizae item)
Instrument: Waters2695 high performance liquid chromatograph, Waters2996 UV-detector
Chromatographic condition and system suitability test are filler with octadecylsilane chemically bonded silica; Methanol-0.2mol/L Spirit of Mindererus .-glacial acetic acid (67: 33: 1) is a mobile phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the monoammonium glycyrrhizinate peak should be not less than 2000.
The preparation of reference substance solution can only compare with existing monoammonium glycyrrhizinate (labelled content 98%) owing to do not bought the standard monoammonium glycyrrhizinate reference substance that Chinese pharmaceutical biological product is identified institute, with definite technology.Therefore, measured result is on the low side, and does not influence test trend.
Gained sample under each extraction conditions in the accurate absorption of the preparation of the need testing solution table 2, water bath method, fixed molten with mobile phase, sample introduction in the range of linearity calculates, promptly.
Factor level table and experimental result are as follows:
Table 1 extraction process of Glycyrrhiza uralensis orthogonal experiment factor level table
Table 2 extraction process of Glycyrrhiza uralensis Orthogonal experiment results
Table 3 Radix Scutellariae extraction process analysis of variance table
| Soruces of variation | Sum of squares between groups | Degree of freedom | Mean square | The F value | Factor affecting |
| A B C error | 0.8283 0.0896 0.1783 0.0334 | 2 2 2 2 | 0.4142 0.0448 0.0892 0.0167 | 24.80 2.68 5.34 | Significantly or not remarkable |
F
0.05(2，2)＝19
F
0.01(2，2)＝99
As above shown in the figure The results of analysis of variance, factor A influence is significant difference and A
2＞A
3＞A
1So select horizontal A
2B in factor B
3＞B
1＞B
2So, select B
3And in factor C C
3＞C
2＞C
1But, C
3With C
2Difference is little, and we select C according to experimental result and document
2Therefore should select A according to experimental result
2B
3C
2Be optimum process condition.But from level difference and variance analysis, it is not remarkable for result's influence that the B factor promptly concentrates density, therefore, from little test result, we select density 1.1 comparatively suitable, and promptly decocting boils three times, amount of water is respectively 10,8,8 times, and the time was respectively 2,1,1 hours; Decocting liquid is concentrated into relative density 1.1 after merging, and alcohol precipitation concentration is 65%.
Two, Pheretima Study on extraction
We adopt new extraction process, extract antiviral therapy and prevention upper respiratory tract infection effective ingredient that fresh Pheretima contains, we find that these compositions are class materials based on the DNA enzyme in prescription screening, so we have adopted biochemical method to carry out the extraction of protease blending constituent.
A. Pheretima Study on Preparation
Pretreatment
Fresh Pheretima is with the dirt of tap water rinsing body surface, puts in 18-22 ℃ the distilled water and soaks 8 hours, and rinsing in per 2 hours is also changed water, makes Pheretima tell earth in the clean gastrointestinal tract.Add 0.i%NaCl then, mixing stimulates it to secrete a large amount of yellow secretions, triplicate, and each 20 minutes, interim was with 10 times of volume distilled water rinsing 2 times.Placing diameter then is the nylon wire of 0.5mm, inhales with powerful water-absorption fiber and removes unnecessary liquid ,-20 ℃ of freezing preservations.
The preparation of crude protein liquid
Get the Pheretima of handling, add acetic acid-sodium-acetate buffer of 5 times of volume 0.1mol/L, mixing was placed 20 hours under the room temperature, foaming, and with industrial refiner 500rpm homogenate 15 minutes, per 3 minutes intermittently 1 minute, homogenized temperature was 20 ℃.Centrifugal 20 minutes of 10000rpm, it is standby to get supernatant.
The preparation of fine straining liquid
Get supernatant and put in 55 ℃ 36 hours, centrifugal 20 minutes of 10000rpm gets supernatant and carries out ultrafiltration dialysis, and outer water dialysis solution is 50 times of volumes of sample, and sample is concentrated 15-18 doubly after the ultrafiltration, is clear and bright colourless or little brown liquid, and is freezing standby.
B. craft screening
Proteic separation key of purifying is processing to raw material, thus we the Pheretima lytic agent after handling in the technology is swashed molten method and direct homogenate after column chromatography carried out comparative study.Concrete grammar is as follows:
Swash molten method
Pheretima after the pretreatment is made the earthworm protein self-dissolving with the acetic acid of 5 times of volume 0.1mol/L of lytic agent-sodium-acetate buffer foaming, carry out homogenate then, supernatant is crude protein solution behind the high speed centrifugation.After crude protein solution prepared fine straining liquid by preceding method, lyophilization was calculated productive rate with the ratio of lyophilized powder and raw material Pheretima.
Column chromatography
With the direct homogenate of dried Pheretima after the pretreatment, then centrifugal back supernatant is crossed post, the albumen eluent is crude protein solution.After crude protein solution prepared fine straining liquid by preceding method, lyophilization was calculated productive rate with the ratio of lyophilized powder and raw material Pheretima.
The experimental result of two kinds of technologies is as follows:
The finished product productive rate of table 4 different process, specific activity are
| Technology | Swash molten method | Column chromatography |
| Productive rate activity yied raising rate gross activity raising rate | 1.218% (83U/mg of Pheretima/Kg) 74% 44.42% | 0.7% (100U/mg of Pheretima/Kg)-- |
From above interpretation of result, swash molten method extraction Pheretima active component product yield and gross activity all are greatly improved.And the sharp molten method of employing not only can be extracted the active component in the raw material to greatest extent, and reduce impurity cleaning and problem of environmental pollution after centrifugal, and direct homogenate can not make protein all dissolve in the column chromatography, loss of effective components is more, and centrifugal back produces a large amount of garbages, the handling problem that has brought a lot of industrial dirts.
Swash molten method and omitted the column chromatography process, not only saved a large amount of production costs, and removed the rate-limiting step of production link, follow-up works such as ultrafiltration have been simplified, enhance productivity greatly, end-product is active simultaneously obviously improves, and is a kind of economical and practical production technology efficiently.
The research of experimental example 3 present composition pharmacological experiments
Used imperial a kind of reed mentioned in ancient books grass spray is present composition spray agent (preparation sees embodiment for details) in the experiment.
1. imperial a kind of reed mentioned in ancient books grass spray is separated the pharmacodynamic study of heat test
Experiment adopts escherichia coli endotoxin to cause Japanese white big ear rabbit fever model, rabbit body temperature begins to rise behind the intravenous injection endotoxin, reached first peak to 1-2 hour, after this body temperature is stable at this or slightly lifting, to thermoregulation secondary rising in about the 4th hour and above first body temperature peak, after this descend gradually, continue more than 8 hours, model group rabbit body temperature curve is the typical double-peak pattern of fever of endotoxin heating.Compare with model group, imperial a kind of reed mentioned in ancient books grass spray has the refrigeration function (P＜0.01) of highly significant, and is obvious dose-effect relationship, and its refrigeration function is better than SHUANGHUANGLIAN KOUFUYE.
2. the pharmacodynamic study that imperial a kind of reed mentioned in ancient books grass spray reduces phlegm and tests
Make the mice atomizing suck phenol red normal saline in the experiment, reflect the secretion capacity of its tunica mucosa tracheae by the phenol red absorbance in the detection mice trachea washing liquid.The result shows that the phenol red excretion amount of imperial a kind of reed mentioned in ancient books grass spray 5.91g/kg group mice obviously increases, the utmost point significant difference (p＜0.01) of having compared with the blank group, and other each group nothings obviously change (P＞0.05).
3. the pharmacodynamic study of imperial a kind of reed mentioned in ancient books grass spray antitussive test
Experiment is adopted mice ammonia to draw and is coughed model, observes imperial a kind of reed mentioned in ancient books grass spray to the mouse cough incubation period of this model and the influence of cough number of times.The result shows that the agent of imperial a kind of reed mentioned in ancient books grass spray influences not quite the cough latent period of ammonia induced mice cough, each group and the blank not statistically significant (P＞0.05) of comparing of organizing; And the cough number of times of imperial a kind of reed mentioned in ancient books grass spray 1.48g/kg group mice obviously reduces, and compares with the blank group, has utmost point significant difference (p＜0.01), and other each group nothings obviously change (P＞0.05).
4. imperial a kind of reed mentioned in ancient books grass spray is to the influence of the effect of the expression of Acute Lung Injury model lung tissue TNF, inflammatory factors such as IL-6, IL-2 due to the oleic acid
The expression of TNF, IL-6, IL-2 is lower in the normal control lung tissue of rats, the expression of interior TNF, IL-6 of acute lung injury rat model lung tissue, IL-2 is obviously increased due to the oleic acid, positive drug group, imperial a kind of reed mentioned in ancient books spray 2.85g/kg group, 0.71g/kg group can obviously reduce the expression of TNF, IL-6, IL-2 in the acute lung injury rat model lung tissue due to the oleic acid, relatively has statistical significance (p＜0.05 with model group, p＜0.01), small dose group reduces not obvious.
5. imperial a kind of reed mentioned in ancient books grass spray minimal bactericidal concentration is measured
Adopt the liquid diluting method to measure imperial a kind of reed mentioned in ancient books grass spray minimal bactericidal concentration.Dragon a kind of reed mentioned in ancient books grass spray is 1: 4 to the minimum sterilization dilution factor of staphylococcus aureus, is 1: 16 to the minimum sterilization dilution factor of beta hemolytic streptococcus, is 1: 2 to the minimum sterilization dilution factor of klepsiella pneumoniae; And escherichia coli and bacillus pyocyaneus are not had bactericidal action.
6. the agent of imperial a kind of reed mentioned in ancient books grass sprayization is to the influence of Acute Lung Injury model due to the oleic acid
Acute Lung Injury model due to the employing oleic acid.Dragon a kind of reed mentioned in ancient books grass spray can effectively promote oleic acid to cause the pulmonary function of acute lung injury rat, arteriotony and heart rate, arterial blood gas, the lung coefficient of surface tension, lung tissue surfactant content, lung coefficient and moisture percentage ratio, indexs such as lung tissue disease's change nature and norphometry thereof all are clearly better, and meet or exceed the Allylnoroxymorphone therapeutical effect.The result proves that the acute lung injury that imperial a kind of reed mentioned in ancient books grass spray causes oleic acid has significant preventive and therapeutic effect.
7. the agent of imperial a kind of reed mentioned in ancient books grass sprayization is to the influence of the chronic rat model of injury of lung due to the monocrotaline
Rat chronic injury of lung model due to the employing monocrotaline, dragon a kind of reed mentioned in ancient books grass spray can significantly promote monocrotaline to cause the pulmonary function of chronic pulmonary injury rats, the pulmonary artery blood pressure, arterial blood gas, lung coefficient of surface tension, lung tissue surfactant content, the lung tissue hydroxyproline content, Serum hyaluronic acid content, the organ coefficient of lung, heart, liver, indexs such as lung tissue pathological changes and collagen fiber deposition and norphometry thereof all take a turn for the better, and the part curative effect is better than nifedipine.Dragon a kind of reed mentioned in ancient books grass spray has significant preventive and therapeutic effect to the chronic pulmonary damage that monocrotaline causes.
8. the pharmacodynamic study of the external resisiting influenza virus of imperial a kind of reed mentioned in ancient books grass spray
Test is adopted to observe mouse lung pathological changes rate, dead protective rate after the administration and to prolong vital rates and is detected in imperial a kind of reed mentioned in ancient books grass spray body inhibitory action to influenza virus; the result shows that imperial a kind of reed mentioned in ancient books grass spray can significantly reduce the mouse invasion rate, alleviates the pulmonary lesion rate and prolong vital rates; learn processing difference by statistics and have significance (p＜0.01), the result shows that imperial a kind of reed mentioned in ancient books grass spray has the effect of remarkable inhibition influenza virus.
9. the external anti-3 type grippe virus of imperial a kind of reed mentioned in ancient books grass spray, respiratory syncytial virus, adenovirus type III, rhinoviral pharmacodynamic study
Adopt virus (CPE) method, in Testis et Pentis Canis passage cell (MDCK), pharyngeal cancer epithelial cell (Hep-2), human embryo kidney (HEK) passage cell (293) and cervical cancer passage cell (HeLa), detect the inhibitory action of the external anti-3 type grippe virus of imperial a kind of reed mentioned in ancient books grass spray, respiratory syncytial virus, adenovirus type III, rhinovirus 14 types, the result shows that imperial a kind of reed mentioned in ancient books grass spray has certain inhibitory action to 3 type grippe virus, respiratory syncytial virus, adenovirus type III, but not obvious to rhinovirus 14 type effects.
Following embodiment all can realize the effect of above-mentioned experimental example.
Embodiment 1 imperial a kind of reed mentioned in ancient books grass spray
Pheretima 300g baicalin 6.5g Radix Glycyrrhizae 300g
More than three the flavor, get Pheretima 300g after the pretreatment, acetic acid-the sodium-acetate buffer that adds 5 times of volume 0.1mol/L, mixing was placed after 20 hours under the room temperature, 20 ℃ of 500rpm homogenate 15 minutes, centrifugal 20 minutes of 10000rpm, getting supernatant put in 55 ℃ 36 hours, centrifugal 20 minutes of 10000rpm gets supernatant and carries out ultrafiltration dialysis, and concentrated solution cold preservation is standby; Radix Glycyrrhizae 300g decocts with water, and the first time, 10 times of water gagings were 2 hours, each 8 times of water gaging of second and third time 1 hour, collecting decoction filters, and filtrate is concentrated into relative density 1.10 (60 ℃ of surveys), add ethanol and make and contain the alcohol amount and reach 65%, left standstill 12 hours, filter, reclaim ethanol to there not being the alcohol flavor, add the Pheretima extracting solution, stir evenly, with ammonia adjust pH 5.5-6.5, add baicalin 6.5g, it is an amount of to add water, stir evenly, cold preservation (4-8 ℃) is spent the night, and filters, add 0.2% sodium benzoate and 0.6% apple essence, stir evenly, filter, add water and make 1000ml, fill, promptly.
[usage and consumption] nasal cavity, mouthspray, each 3-5 presses, every day 6-12 time or abide by testament.
[specification] 25ml/ bottle
Embodiment 2 oral liquids
Pheretima 400g baicalin 8.5g Radix Glycyrrhizae 400g
More than three the flavor, get Pheretima 400g after the pretreatment, acetic acid-the sodium-acetate buffer that adds 6 times of volume 0.1mol/L, mixing was placed after 24 hours under the room temperature, 24 ℃ of 500rpm homogenate 15 minutes, centrifugal 20 minutes of 10000rpm, getting supernatant put in 55 ℃ 36 hours, centrifugal 20 minutes of 10000rpm gets supernatant and carries out ultrafiltration dialysis, and concentrated solution cold preservation is standby; Radix Glycyrrhizae 400g decocts with water, and the first time, 12 times of water gagings were 2 hours, each 8 times of water gaging of second and third time 1 hour, collecting decoction, filter, filtrate is concentrated into relative density 1.10 (60 ℃ of surveys), adds ethanol and makes and contain the alcohol amount and reach 65%, leaves standstill 12 hours, filter, reclaim ethanol, add the Pheretima extracting solution to there not being the alcohol flavor, stir evenly,, add baicalin 8.5g with ammonia adjust pH 5.5--6.5, it is an amount of to add water, stirs evenly cold preservation (4-8 ℃), spend the night, filter, make oral liquid 1000ml. through conventional technology
Embodiment 3 tablets
Pheretima 250g baicalin 4.5g Radix Glycyrrhizae 300g
More than three the flavor, get Pheretima 250g after the pretreatment, acetic acid-the sodium-acetate buffer that adds 4 times of volume 0.1mol/L, mixing was placed after 20 hours under the room temperature, 20 ℃ of 500rpm homogenate 15 minutes, centrifugal 20 minutes of 10000rpm, getting supernatant put in 55 ℃ 36 hours, centrifugal 20 minutes of 10000rpm gets supernatant and carries out ultrafiltration dialysis, and concentrated solution cold preservation is standby; Radix Glycyrrhizae 300g decocts with water, and the first time, 9 times of water gagings were 1.5 hours, each 9 times of water gaging of second and third time 1.5 hours, collecting decoction, filter, filtrate is concentrated into relative density 1.10 (60 ℃ of surveys), adds ethanol and makes and contain the alcohol amount and reach 65%, leaves standstill 12 hours, filter, reclaim ethanol, add the Pheretima extracting solution to there not being the alcohol flavor, stir evenly,, add baicalin 4.5g with ammonia adjust pH 5.5--6.5, it is an amount of to add water, stirs evenly cold preservation (4-8 ℃), spend the night, drying is made tablet through conventional technology.
Embodiment 4 capsules
Pheretima 350g baicalin 6.5g Radix Glycyrrhizae 250g
More than three the flavor, get Pheretima 350g after the pretreatment, acetic acid-the sodium-acetate buffer that adds 9 times of volume 0.1mol/L, mixing was placed after 20 hours under the room temperature, 20 ℃ of 500rpm homogenate 15 minutes, centrifugal 20 minutes of 10000rpm, getting supernatant put in 55 ℃ 36 hours, centrifugal 20 minutes of 10000rpm gets supernatant and carries out ultrafiltration dialysis, and concentrated solution cold preservation is standby; Radix Glycyrrhizae 250g decocts with water, and the first time, 10 times of water gagings were 2 hours, each 8 times of water gaging of second and third time 1 hour, collecting decoction, filter, filtrate is concentrated into relative density 1.10 (60 ℃ of surveys), adds ethanol and makes and contain the alcohol amount and reach 65%, leaves standstill 12 hours, filter, reclaim ethanol, add the Pheretima extracting solution to there not being the alcohol flavor, stir evenly,, add baicalin 6.5g with ammonia adjust pH 5.5-6.5, it is an amount of to add water, stirs evenly cold preservation (4-8 ℃), spend the night, drying is made capsule through conventional technology.
Embodiment 5 imperial a kind of reed mentioned in ancient books grass spray method of quality control
Differentiate:
1. Radix Scutellariae: get this product liquid 10ml, water bath method, residue add methanol 5ml thermosol, are concentrated into 1ml, as need testing solution.Other gets the baicalin reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography (appendix VIB), draw each 5 μ l of above-mentioned two kinds of solution, put respectively in same be on the silica gel g thin-layer plate of adhesive with the carboxymethylcellulose sodium solution, with 5: 3: 1: 1 ethyl acetate-butanone-formic acid-water developing solvent, pre-equilibration 30 minutes launches, and takes out, dry, spray is with 1% ferric chloride alcoholic solution; In the test sample chromatograph with baicalin reference substance chromatograph relevant position on show an identical dirty-green speckle;
2. Radix Glycyrrhizae: get this product liquid 10ml, add methanol 50ml reflux, extract, 1 hour, reclaim methanol to doing, add 30ml water and make dissolving, with n-butanol extraction 3 times, each 20ml merges n-butyl alcohol liquid, and evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution; Extracting liquorice control medicinal material 0.5g in addition, with the test sample method operate solution 0.5ml, medical material solution in contrast.Test according to thin layer chromatography (appendix VIB), draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same prefabricated silica gel g thin-layer plate with the 1%NaOH solution spray, with 15: 1: 1: 2 ethyl acetates-formic acid-glacial acetic acid-water developing solvent, launch, take out, dry, spray is put under the 365nm ultra-violet lamp and is inspected with 10% ethanol solution of sulfuric acid; In the test sample chromatograph with control medicinal material chromatograph relevant position on show the fluorescence speckle of a same color;
3. Pheretima: get this product 10ml, add chloroform 20ml, close plug, supersound process 20 minutes filters, and filtrate is concentrated into about 1ml, as need testing solution.Other gets Pheretima medical material 3g, makes medical material solution with method; According to thin layer chromatography (appendix VIB) test, draw each 5 μ l of above-mentioned two clock solution, put respectively on same silica gel g thin-layer plate, be developing solvent with 9: 1 benzene one ethyl acetates, launch, take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of medical material chromatograph on, show the fluorescence speckle of same color.
Assay:
Baicalin: measure chromatographic condition and system suitability test according to high performance liquid chromatography (appendix VID of Chinese Pharmacopoeia version in 2000): with octadecylsilane chemically bonded silica is filler; 47: 53: 0.2 methanol-water-phosphoric acid are mobile phase; The detection wavelength is 280nm; Number of theoretical plate calculates by the baicalin peak should be not less than 2500; The preparation of reference substance solution: precision takes by weighing at 4 hours baicalin reference substance of 60 ℃ of drying under reduced pressure an amount of, adds methanol and makes the solution that every 1ml contains 64.40 μ g.The preparation of need testing solution: precision is measured this product solution 1ml and is put in the 100ml measuring bottle, adds methanol to scale, shakes up, and puts slight fever in the water-bath, promptly; Algoscopy: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject hplc determination, external standard method is calculated, promptly; The every 25ml liquid of this product contains baicalin (C
21H
18O
11) must not be less than 67.5mg.
Claims (10)
1, a kind of Chinese medicine composition is characterized in that said composition made by following bulk drugs:
Pheretima 100-500 weight portion baicalin 1-10 weight portion Radix Glycyrrhizae 100-500 weight portion.
2, Chinese medicine composition as claimed in claim 1 is characterized in that said composition made by following bulk drugs:
Pheretima 200-400 weight portion baicalin 3-8 weight portion Radix Glycyrrhizae 200-400 weight portion.
3, Chinese medicine composition as claimed in claim 2 is characterized in that said composition made by following bulk drugs:
Pheretima 300 weight portion baicalins 6.5 weight portion Radix Glycyrrhizaes 300 weight portions.
4, Chinese medicine composition as claimed in claim 2 is characterized in that said composition made by following bulk drugs:
Pheretima 400 weight portion baicalins 8.5 weight portion Radix Glycyrrhizaes 400 weight portions.
5, a kind of Chinese medical spray preparation is characterized in that this spray is mainly to be made by following bulk drugs:
Pheretima 100-500 weight portion baicalin 1-10 weight portion Radix Glycyrrhizae 100-500 weight portion.
6, spray agent as claimed in claim 5 is characterized in that this spray is mainly to be made by following bulk drugs:
Pheretima 200-400 weight portion baicalin 3-8 weight portion Radix Glycyrrhizae 200-400 weight portion.
7, spray agent as claimed in claim 6 is characterized in that this spray is mainly to be made by following bulk drugs:
Pheretima 300 weight portion baicalins 6.5 weight portion Radix Glycyrrhizaes 300 weight portions.
8,, it is characterized in that said composition directly or indirectly adds pharmaceutically acceptable excipient through conventional operation and makes clinical tablet, capsule, granule, drop pill, oral liquid as the described Chinese medicine composition of claim 1-4.
9,, be characterised in that said composition is used for suppressing the application of influenza virus, upper respiratory tract infection virus or SARS virus and antibacterial medicine in preparation as the described Chinese medicine composition of claim 1-4.
10,, be characterised in that said composition is used for suppressing the application of influenza virus, upper respiratory tract infection virus or SARS virus and antibacterial medicine in preparation as the described spray of claim 5-7.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101837046A (en) * | 2010-05-27 | 2010-09-22 | 辽宁中医药大学 | Anti-virus Chinese medicinal preparation and preparation method thereof |
| CN102236006A (en) * | 2011-06-24 | 2011-11-09 | 大连美罗中药厂有限公司 | Method for detecting earthworm in infantile lung-heat clearing granules |
| CN104758362A (en) * | 2015-04-26 | 2015-07-08 | 淄博齐鼎立专利信息咨询有限公司 | Application of Chinese traditional medicine composition in preparing medicine for resisting respiratory syncytial viruses |
| CN109260231A (en) * | 2017-07-18 | 2019-01-25 | 首都儿科研究所 | The preparation method of the anti-inflammatory antimicrobial extract of cough-relieving apophlegmatic in a kind of earthworm |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1054752C (en) * | 1994-10-10 | 2000-07-26 | 陈昌陶 | Anti-virus and antipyretic oral liquid |
| CN1364538A (en) * | 2001-01-11 | 2002-08-21 | 杨孟君 | Nano bronchitis treating medicine and its preparing method |
| CN1201793C (en) * | 2003-04-08 | 2005-05-18 | 陈杰元 | Medicine composition for treatment of cough and asthma and preparation thereof |
| CN1207301C (en) * | 2003-05-25 | 2005-06-22 | 韩桂茹 | Process of extracting baicalin from scutellaria baicalensis |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101837046A (en) * | 2010-05-27 | 2010-09-22 | 辽宁中医药大学 | Anti-virus Chinese medicinal preparation and preparation method thereof |
| CN102236006A (en) * | 2011-06-24 | 2011-11-09 | 大连美罗中药厂有限公司 | Method for detecting earthworm in infantile lung-heat clearing granules |
| CN104758362A (en) * | 2015-04-26 | 2015-07-08 | 淄博齐鼎立专利信息咨询有限公司 | Application of Chinese traditional medicine composition in preparing medicine for resisting respiratory syncytial viruses |
| CN109260231A (en) * | 2017-07-18 | 2019-01-25 | 首都儿科研究所 | The preparation method of the anti-inflammatory antimicrobial extract of cough-relieving apophlegmatic in a kind of earthworm |
| CN109260231B (en) * | 2017-07-18 | 2021-09-03 | 首都儿科研究所 | Preparation method of earthworm extract with cough stopping, phlegm eliminating, anti-inflammatory and antimicrobial functions |
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