CN101926950B - Brain-invigorating traditional Chinese medicine composite and preparation method and detection method thereof - Google Patents
Brain-invigorating traditional Chinese medicine composite and preparation method and detection method thereof Download PDFInfo
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Abstract
The invention relates to a brain-invigorating traditional Chinese medicine composite and a preparation method thereof. The traditional Chinese medicine composite of the invention comprises the traditional Chinese medicine raw materials in parts by weight: 20-40 parts of Chinese angelica, 5-15 parts of Tabasheer, 15-35 parts of cistanche, 5-15 parts of dens draconis, 15-35 parts of Chinese yam, 5-15 parts of amber, 10-25 parts of Schisandra chinensis, 3-10 parts of gastrodia elata, 2-8 parts of semen boitae, 3-10 parts of Salvia miltiorrhiza, 10-25 parts of fructus alpiniae oxyphyllae, 3-10 parts of ginseng, 5-15 parts of polygala tenuifolia, 3-10 parts of chrysanthemum, 5-15 parts of Irkutsk Anemone Rhizome, 5-15 parts of ochre, 5-15 parts of arisaema cum bile, 35-60 parts of spina date seed and 15-30 parts of barbary wolfberry fruit. The invention also provides a detection method for the composite according to the components of the traditional Chinese medicine composite. The traditional Chinese medicine composite of the invention has good brain-invigorating effect. The animal testing proves that the traditional Chinese medicine composite of the invention has the functions of accelerating sleep, resisting fatigue and strengthening humoral immunity, can be used for invigorating brain, improving intellectual development, nourishing heart and soothing the nerves, and can be used for treating brain overstrain, fading memory, neurasthenia, light-headedness, fright insomnia, feeling of oppression over the chest, easy tiredness and Senile Dementia. The detection method has good ruggedness and high sensitivity.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition and preparation method thereof and detection method, particularly Chinese medicine composition of a kind of nourishing the brain and improving intelligence and preparation method thereof and detection method.
Background technology
Along with development of modern society, have the medicine of brain-invigorating and attention and the favor that food more and more receives people.At present, the brain-strengthening drug variety is various, but curative effect is unsatisfactory, and most medicines need be taken the long time, could be slightly effective in cure.
It is little that China's Chinese medicine has side effect, and the characteristics that conditioning functions is outstanding are taken the course of its own in the brain-strengthening field.
The present invention combine pharmacy, pharmacodynamics and the theory of Chinese medical science of modern medicine aim to provide a kind of fast and effectively, have nourishing the brain and improving intelligence, the effects such as mending body of sleeping peacefully, can be used for brain overwork, amnesia, neurasthenia, the insomnia of having a dizzy spell, palpitate with fear, vexedly be prone to tired, fear of cold body void, body loses the Chinese medicine composition of diseases such as lumbago and senile dementia.
Summary of the invention
The present invention gets purpose and is to provide a kind of brain-strengthening Chinese medicine composition, and another object of the present invention is to provide this pharmaceutical composition to get method for preparing and detection method.
The present invention seeks to realize through following technical scheme:
Chinese medicine composition of the present invention comprises following parts by weight of Chinese traditional medicine crude drug:
Radix Angelicae Sinensis: 20-40; Concretio Silicea Bambusae: 5-15;
Herba Cistanches: 15-35; Dens Draconis: 5-15;
Rhizoma Dioscoreae: 15-35; Succinum: 5-15;
Fructus Schisandrae Chinensis: 10-25; Rhizoma Gastrodiae: 3-10;
Semen Platycladi: 2-8; Radix Salviae Miltiorrhizae: 3-10;
Fructus Alpiniae Oxyphyllae: 10-25; Radix Ginseng: 3-10;
Radix Polygalae: 5-15; Flos Chrysanthemi: 3-10;
Rhizoma Anemones Altaicae: 5-15; Haematitum: 5-15;
Arisaema Cum Bile: 5-15; Semen Ziziphi Spinosae: 35-60;
Fructus Lycii: 15-30.
Wherein, said Herba Cistanches is the salt Herba Cistanches (processed); Said Dens Draconis is a calcined DENS DRACONIS; Said Fructus Schisandrae Chinensis is the wine Fructus Schisandrae Chinensis (steamed); Said Semen Platycladi is for frying Semen Platycladi; Said Fructus Alpiniae Oxyphyllae is the salt stir-baked FRUCTUS ALPINIAE OXY-PHYLLAE; Said Radix Polygalae is the liquorice beverage Cortex et Radix Polygalae (processed); Said Semen Ziziphi Spinosae is a Semen Ziziphi Spinosae (parched);
The present invention proposes especially, and Fructus Schisandrae Chinensis according to the invention is the dry mature fruit of magnoliaceae schisandra, and practise and claim " Radix Schisandrae Bicoloris ", but not " Fructus Schisandrae Sphenantherae "; Said Rhizoma Anemones Altaicae is adopted " Shandong Province's Chinese medicine processing standard 2002 " (" Shandong Province's Chinese medicine processing standard " (version in 2002) Shandong Province Drug Administration compiles, Shandong friendship publishing house, the concocting method of the 12nd page of Rhizoma Anemones Altaicae), concocts.
Because the active ingredient of Fructus Schisandrae Sphenantherae and Radix Schisandrae Bicoloris has difference, the preferred Radix Schisandrae Bicoloris of the present invention, the crude drug source of Rhizoma Anemones Altaicae is also relatively more chaotic simultaneously, and is more reliable and more stable for the curative effect that makes Chinese medicine composition of the present invention, and the Chinese medicine that adopts has been carried out standard.
The card that prescription according to the invention cures mainly is that Chinese Angelica blood replonishing is aided with Rhizoma Dioscoreae, Radix Salviae Miltiorrhizae to make strengthening spleen and nourishing blood, blood circulation promoting and enriching by due to the heart kidney yin blood deficiency damage; The Fructus Lycii kidney-yang nourishing, Herba Cistanches temperature and not dry, the kidney invigorating and essence nourishing.In the side, Radix Ginseng QI invigorating mind calming; Semen Ziziphi Spinosae, Fructus Schisandrae Chinensis, Dens Draconis the is sour and astringent convergence motive are with tranquilizing mind; Allay excitement in succinum, Haematitum important city; Semen Platycladi, Radix Polygalae, Fructus Alpiniae Oxyphyllae the heart nourishing and intellengence improving; Rhizoma Anemones Altaicae, Rhizoma Gastrodiae, Concretio Silicea Bambusae, Arisaema Cum Bile, Flos Chrysanthemi YANG hyperactivity suppressing clear away heart-fire.This prescription heart kidney giving young employees remedial-courses in general knowledge and vocational skills can be treated deficiency of YIN-blood originally, the forgetful index of sleeping less, and treating both the principal and secondary aspects of a disease, natural brain-strengthening is opened intelligence, mind tranquilizing and the heart calming.
Preferably, Chinese medicine composition of the present invention comprises following parts by weight of Chinese traditional medicine crude drug:
Radix Angelicae Sinensis: 20-35; Zhu is yellow: 10-15;
Herba Cistanches: 20-27; Dens Draconis: 10-15;
Rhizoma Dioscoreae: 20-27; Succinum: 10-15;
Fructus Schisandrae Chinensis: 15-21; Rhizoma Gastrodiae: 5-8;
Semen Platycladi: 4-6; Radix Salviae Miltiorrhizae: 5-8;
Fructus Alpiniae Oxyphyllae: 15-21; Radix Ginseng: 5-8;
Radix Polygalae: 10-15; Flos Chrysanthemi: 5-8;
Rhizoma Anemones Altaicae: 10-15; Haematitum: 7-11;
Arisaema Cum Bile: 10-15; Semen Ziziphi Spinosae: 40-55;
Fructus Lycii: 20-27.
Special, the preferred following two kinds of Chinese medicine compositions of the present invention:
1) comprise the Chinese medicine composition of following parts by weight of Chinese traditional medicine crude drug:
Radix Angelicae Sinensis: 33.3; Concretio Silicea Bambusae: 13.3;
Herba Cistanches (salt is processed): 26.7; Dens Draconis (forging): 13.3;
Rhizoma Dioscoreae: 26.7; Succinum: 13.3;
Fructus Schisandrae Chinensis (wine steaming): 20; Rhizoma Gastrodiae: 6.7;
Semen Platycladi (stir-fry): 5.3; Radix Salviae Miltiorrhizae: 6.7;
Fructus Alpiniae Oxyphyllae (salt stir-fry): 20; Radix Ginseng: 6.7;
Radix Polygalae (liquorice beverage is processed): 13.3; Flos Chrysanthemi: 6.7;
Rhizoma Anemones Altaicae: 13.3; Haematitum: 10;
Arisaema Cum Bile: 13.3; Semen Ziziphi Spinosae (stir-fry): 53.3;
Fructus Lycii: 26.7.
2) and the Chinese medicine composition that comprises following parts by weight of Chinese traditional medicine crude drug:
Radix Angelicae Sinensis: 25; Concretio Silicea Bambusae: 10;
Herba Cistanches (salt is processed): 20; Dens Draconis (forging): 10;
Rhizoma Dioscoreae: 20; Succinum: 10;
Fructus Schisandrae Chinensis (wine steaming): 15; Rhizoma Gastrodiae: 5;
Semen Platycladi (stir-fry): 4; Radix Salviae Miltiorrhizae: 5;
Fructus Alpiniae Oxyphyllae (salt stir-fry): 15; Radix Ginseng: 5;
Radix Polygalae (liquorice beverage is processed): 10; Flos Chrysanthemi: 5;
Rhizoma Anemones Altaicae: 10; Haematitum: 7.5;
Arisaema Cum Bile: 10; Semen Ziziphi Spinosae (stir-fry): 40;
Fructus Lycii: 20.
Compositions of the present invention is through studying and adjust the consumption of prescription taste of Chinese medicine, more than 2 prescription proportionings demonstrate beyond thought effect, with prior art relatively, significant effect is arranged aspect pharmacodynamics.
Chinese medicine composition of the present invention can adopt the known technology of this area to process various dosage forms, like pharmaceutically acceptable dosage forms such as tablet, pill, powder, capsule, granule and liquid agent: preferred capsule, pill or tablet.
The method for preparing of Chinese medicine composition of the present invention is: get the raw material of Chinese medicine medicine; Haematitum, succinum, Concretio Silicea Bambusae are ground into fine powder respectively, and all the other ten Six-elements are ground into fine powder, with above-mentioned fine powder facing-up; Sieve; Mixing is prepared into the acceptable various dosage forms of pharmacy by technology well known in the art then, as processes capsule, pill, powder, granule or tablet.
In addition; The method for preparing of the pill of Chinese medicine composition of the present invention is: with Haematitum, succinum, the single porphyrize powder of Concretio Silicea Bambusae difference, all the other ten Six-elements are ground into fine powder, sieve with 30%-60% Haematitum, succinum, Concretio Silicea Bambusae fine powder again; Mixing; Add the sodium alginate mixing with water pill or machine pill, polishing, drying; With Resina persicae, residue Haematitum coating, drying is processed pill; Preferred earlier with 50% Haematitum, succinum, Concretio Silicea Bambusae fine powder and other ten Six-elements mixing pills, then 50% Haematitum is used for coating.
Special, the present invention proposes the method for preparing of above-mentioned Chinese medicinal composition capsules, get the raw material of Chinese medicine medicine of following weight:
Radix Angelicae Sinensis: 33.3g; Concretio Silicea Bambusae: 13.3g;
Herba Cistanches (salt is processed): 26.7g; Dens Draconis (forging): 13.3g;
Rhizoma Dioscoreae: 26.7g; Succinum: 13.3g;
Fructus Schisandrae Chinensis (wine steaming): 20g; Rhizoma Gastrodiae: 6.7g;
Semen Platycladi (stir-fry): 5.3g; Radix Salviae Miltiorrhizae: 6.7g;
Fructus Alpiniae Oxyphyllae (salt stir-fry): 20g; Radix Ginseng: 6.7g;
Radix Polygalae (liquorice beverage is processed): 13.3g; Flos Chrysanthemi: 6.7g;
Rhizoma Anemones Altaicae: 13.3g; Haematitum: 10g;
Arisaema Cum Bile: 13.3g; Semen Ziziphi Spinosae (stir-fry): 53.3g;
Fructus Lycii: 26.7g;
Haematitum, succinum, Concretio Silicea Bambusae are ground into fine powder respectively, and all the other ten Six-elements are ground into fine powder, with above-mentioned fine powder facing-up, sieve, and mixing, the system granule, drying, encapsulated, process 1000.
The invention allows for the method for preparing of above-mentioned Chinese medicine composition pill, get the Chinese medicine composition that comprises following parts by weight of Chinese traditional medicine crude drug:
Radix Angelicae Sinensis: 25; Concretio Silicea Bambusae: 10;
Herba Cistanches (salt is processed): 20; Dens Draconis (forging): 10;
Rhizoma Dioscoreae: 20; Succinum: 10;
Fructus Schisandrae Chinensis (wine steaming): 15; Rhizoma Gastrodiae: 5;
Semen Platycladi (stir-fry): 4; Radix Salviae Miltiorrhizae: 5;
Fructus Alpiniae Oxyphyllae (salt stir-fry): 15; Radix Ginseng: 5;
Radix Polygalae (liquorice beverage is processed): 10; Flos Chrysanthemi: 5;
Rhizoma Anemones Altaicae: 10; Haematitum: 7.5;
Arisaema Cum Bile: 10; Semen Ziziphi Spinosae (stir-fry): 40;
Fructus Lycii: 20.
With Haematitum, succinum, the single porphyrize powder of Concretio Silicea Bambusae difference, all the other ten Six-elements are ground into fine powder, sieve with 50% Haematitum, succinum, Concretio Silicea Bambusae fine powder again, and mixing adds the sodium alginate mixing with water pill or machine pill, polishing, drying; With Resina persicae, residue Haematitum coating, drying is processed pill.
But Chinese medicine composition nourishing the brain and improving intelligence of the present invention, tranquilizing by nourishing the heart is used for brain overwork, hypomnesis, the neurasthenia has a dizzy spell, the palpitation with fear insomnia, vexed easy to be tired, the fear of cold body is empty, and body loses diseases such as lumbago and senile dementia.
Compositions of the present invention aspect pharmacy with immediate compared with techniques, have following advantage: resistance to water soak is better, is beneficial to storage, quality is more stable; Significant effect is also arranged aspect pharmacodynamics in addition.
The consumption of Chinese medicine composition of the present invention was generally 0.1-20g/ days, preferred 1-3g/ days, looked crowd's the decision that needs.
The inventor detects each key component of Chinese medicine composition, has proposed the detection method of said Chinese medicine composition, and said detection method comprises discrimination method and content assaying method.
Existing discrimination method comprises the thin layer liquid chromatograph of microscopy and Radix Angelicae Sinensis, specifically, differentiates as follows:
(1) get these article, put microscopically and observe: plant skin palisade cells brownish red, norma lateralis is the string palisade to be arranged, and polygon is seen on the surface, and cell is big slightly, includes the dark-brown thing; The pale brown color of endotesta cell, surface see rectangle or class is square, and the anticlinal wall beaded thickens (Semen Ziziphi Spinosae).Plant the pale brown color of skin epidermis stone cell, a type polygon is seen on the surface, and wall is thicker, and the hole ditch is fine and closely woven, and cell contains burgundy thing (Fructus Schisandrae Chinensis).Plant skin stone cell surface and see irregular polygon, wall thickness, wavy bending, laminated striation clear (Fructus Lycii).
(2) get these article 4g, porphyrize adds ethanol 40ml, and supersound process 20 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution.Other gets Radix Angelicae Sinensis control medicinal material 1g, adds ethanol 10ml, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VI B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with cyclohexane extraction-ethyl acetate (9: 1), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
In addition; The inventor proposes to be applicable to the discrimination method of Chinese medicine composition of the present invention through experimentation, is respectively Fructus Lycii, Herba Cistanches, Fructus Schisandrae Chinensis, Semen Ziziphi Spinosae and Radix Ginseng; Experimentation is found; The separating effect of said medicine in Chinese medicine composition is better, and feminine gender do not see interference, is highly suitable for differentiating Chinese medicine composition of the present invention.
For Chinese medicine composition of the present invention, its discrimination method can also comprise following one or more detection methods, and concrete grammar is following:
(3) sample thief porphyrize, decocting in water is put coldly, and supernatant is got in slagging-off, with ethyl acetate extraction 1-3 time, 20-60ml at every turn, combined ethyl acetate liquid, evaporate to dryness, residue adds acetic acid ethyl dissolution, as need testing solution (every 2.5g sample ligand is set to the 1ml need testing solution); Other gets the Fructus Lycii control medicinal material, shines medical material solution (every 0.5g control medicinal material is configured to the 1ml need testing solution) in pairs with legal system; According to thin layer chromatography (appendix VI B) test, draw each 1-10 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate; Launch with developing solvent, take out, dry; Put under the ultra-violet lamp and inspect, said developing solvent is selected from following a kind of: ethyl acetate-chloroform-formic acid (3: 2: 1); Ethyl acetate-chloroform-formic acid (1: 2: 1); Toluene-ethyl acetate-formic acid (6: 4: 0.2); Ethyl acetate-chloroform-glacial acetic acid (25: 10: 1);
(4) sample thief porphyrize, the supersound process that adds diethyl ether is put coldly, filters, and discards ether solution; Residue volatilizes solvent, adds the methanol supersound process, puts coldly, filters; The filtrating evaporate to dryness, residue adds the water slight fever makes dissolving, through the absorption of D101 macroporous adsorptive resins, eluting; Collect eluent, evaporate to dryness, residue adds dissolve with methanol, as need testing solution (every 5g sample ligand is set to the 1ml need testing solution); Other gets the echinacoside reference substance, adds the solution that methanol is processed 1mg/ml, as reference substance solution; According to thin layer chromatography (appendix VI B) test, draw each 1-5 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, be that developing solvent launches with methanol-acetic acid-water (2: 1: 7), take out, dry, put under the ultra-violet lamp and inspect;
(5) sample thief porphyrize adds the ethanol supersound process, filters, and concentrated filtrate is as need testing solution (every 2.0g sample ligand is set to the 1ml need testing solution); Get the Fructus Schisandrae Chinensis control medicinal material and add the ethanol supersound process, filter, concentrated filtrate is as control medicinal material solution (every 0.5g control medicinal material is configured to the 1ml need testing solution); According to thin layer chromatography (appendix VI B) test, draw each 5-10 μ l of need testing solution and above-mentioned control medicinal material solution respectively, put respectively on same silica GF254 lamellae, launch with developing solvent, take out, dry, put under the ultra-violet lamp and inspect; Said developing solvent is the upper solution of petroleum ether-ethyl acetate-formic acid (15: 5: 1); Or cyclohexane extraction-ethyl acetate (7: 2.5);
(6) get need testing solution under the discrimination method (4), evaporate to dryness, residue is dissolved in water; Extract 1-3 time with water saturated n-butyl alcohol jolting, each 5-20ml merges n-butyl alcohol liquid; With ammonia solution washing 1-3 time, each 5-20ml, n-butyl alcohol liquid evaporate to dryness; Residue adds dissolve with methanol, as need testing solution (every 10g sample ligand is set to the 1ml need testing solution); Other gets jujuboside A and jujuboside B reference substance, adds methanol and processes the mixed solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography (appendix VI B) test, draw above-mentioned reference substance solution and need testing solution 5-10 μ l, put respectively on same silica gel g thin-layer plate; With water saturated n-butyl alcohol is developing solvent, launches, and takes out; Dry, spray is inspected with 2% vanillin sulfuric acid solution immediately;
(7) method according to discrimination method (6) prepares need testing solution; Get the Radix Ginseng control medicinal material, add the methanol supersound process, put cold; Filter, the filtrating evaporate to dryness, residue adds the water slight fever makes dissolving; Through the absorption of D101 macroporous adsorptive resins, eluting is collected eluent; Evaporate to dryness, residue adds dissolve with methanol, as control medicinal material solution (every 0.5g reference substance medical material is configured to the 1ml reference substance solution); According to thin layer chromatography (appendix VI B) test, draw need testing solution and each 1-10 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate; With 10 ℃ of lower floor's solution with held of chloroform-methanol-water (13: 7: 2) is developing solvent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.
Wherein, in the authentication method (5), can also (or) get schisandrin B, schisandrin reference substance simultaneously, get the schisandrin B reference substance, the schisandrin reference substance adds methanol respectively and processes the solution that every 1ml contains 1mg, as reference substance solution as these article.
In the authentication method (7), can also (or simultaneously) get the reference substance of ginsenoside Rg1, get ginsenoside Rg1's reference substance, add methanol and process the solution that every 1ml contains 1mg, as reference substance solution as these article.
In authentication method (3), developing solvent is preferably toluene-ethyl acetate-formic acid of 6: 4: 0.2.
In authentication method (5), developing solvent is preferably 7: 2.5 cyclohexane extraction-ethyl acetate solutions.
Preferably, the discrimination method of Chinese medicine composition of the present invention is:
(1) get these article, put microscopically and observe: plant skin palisade cells brownish red, norma lateralis is the string palisade to be arranged, and polygon is seen on the surface, and cell is big slightly, includes the dark-brown thing; The pale brown color of endotesta cell, surface see rectangle or class is square, and the anticlinal wall beaded thickens (Semen Ziziphi Spinosae).Plant the pale brown color of skin epidermis stone cell, a type polygon is seen on the surface, and wall is thicker, and the hole ditch is fine and closely woven, and cell contains burgundy thing (Fructus Schisandrae Chinensis).Plant skin stone cell surface and see irregular polygon, wall thickness, wavy bending, laminated striation clear (Fructus Lycii).
(2) get these article 4g, porphyrize adds ethanol 40ml, and supersound process 20 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution.Other gets Radix Angelicae Sinensis control medicinal material 1g, adds ethanol 10ml, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VI B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with cyclohexane extraction-ethyl acetate (9: 1), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.
(3) get these article 5g, porphyrize adds water 100ml, and heated and boiled 15 minutes is put coldly, centrifugal, gets supernatant, adds ethyl acetate extraction 3 times, each 50ml, and combined ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate 2ml makes dissolving, as need testing solution.Other gets Fructus Lycii control medicinal material 1g, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VI B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with toluene-ethyl acetate-formic acid (6: 4: 0.2), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.
(4) get these article 10g, porphyrize, the 60ml that adds diethyl ether, supersound process 30 minutes is put cold; Filter, discard ether solution, residue volatilizes solvent, adds methanol 100ml, supersound process 30 minutes; Put coldly, filter, the filtrating evaporate to dryness, residue adds water 10ml slight fever makes dissolving, through D101 macroporous adsorptive resins (internal diameter 1.5cm; Long 15cm), water 70ml eluting discards eluent, and reuse 20% ethanol 70ml eluting discards eluent; Continue with 60% methanol 100ml eluting, collect eluent, evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.Other gets the echinacoside reference substance, adds methanol and processes the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography (appendix VI B) test, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, be developing solvent with methanol-acetic acid-water (2: 1: 7), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.
(5) get Fructus Schisandrae Chinensis control medicinal material 1g, add ethanol 10ml, supersound process 20 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution.Get schisandrin B reference substance, schisandrin reference substance again, add methanol respectively and process the solution that every 1ml contains 1mg, as reference substance solution.Test according to thin layer chromatography (appendix VI B); Drawing need testing solution 10 μ l and above-mentioned control medicinal material solution and each 5 μ l of reference substance solution under the item of [discriminating] (2), put respectively on same silica GF254 lamellae, is developing solvent with cyclohexane extraction-ethyl acetate (7: 2.5); Launch; Take out, dry, put under the ultra-violet lamp (254nm) and inspect.
(6) get discriminating (4) need testing solution down, evaporate to dryness, residue add water 15ml makes dissolving, with water saturated n-butyl alcohol jolting extraction 3 times; Each 15ml merges n-butyl alcohol liquid, with ammonia solution washing 2 times, and each 15ml; N-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets jujuboside A and jujuboside B reference substance, adds methanol and processes the mixed solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography (appendix VI B) test, draw above-mentioned need testing solution 10 μ l and reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate; With water saturated n-butyl alcohol is developing solvent, launches, and takes out; Dry, spray is inspected with 2% vanillin sulfuric acid solution immediately.
(7) get Radix Ginseng control medicinal material 1g, add methanol 50ml, supersound process 30 minutes is put coldly, filters; The filtrating evaporate to dryness, residue adds water 10ml slight fever makes dissolving, through D101 macroporous adsorptive resins (internal diameter 1.5cm, long 15cm), water 70ml eluting; Abandon eluent, reuse 20% ethanol 70ml eluting discards eluent, continues with 60% methanol 100ml eluting; Collect eluent, evaporate to dryness, residue add methanol 2ml makes dissolving, as control medicinal material solution.Other gets ginsenoside Rg1's reference substance, adds methanol and processes the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography (appendix VI B) test, draw need testing solution 10 μ l and above-mentioned control medicinal material solution and each 5 μ l of reference substance solution of differentiating under (6) item, put respectively on same silica gel g thin-layer plate; With 10 ℃ of lower floor's solution with held of chloroform-methanol-water (13: 7: 2) is developing solvent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.
The invention allows for the detection method of content of this Chinese medicine composition, said detection method of content is a detection method of measuring the Fructus Schisandrae Chinensis active constituent content, comprising:
1) preparation of need testing solution: the sample porphyrize, the accurate title, decide, and adds methanol, supersound process; Filter, discard filtrating just, get the subsequent filtrate evaporate to dryness, residue adds dissolve with methanol; Shake up, filter, get subsequent filtrate, promptly get (0.5-1.5g sample configuration 10ml sample solution) with microporous filter membrane.
2) preparation of reference substance solution: get the schisandrin reference substance, add methanol and process the solution that every 1ml contains 25 μ g, promptly get;
3) measure: draw each 1-20 μ l of reference substance solution and need testing solution respectively, inject chromatograph of liquid, measure.
Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; With methanol-water (53: 47) is mobile phase; The detection wavelength is 250nm; Number of theoretical plate calculates by the schisandrin peak and is not less than 3000.
Preferably, the content assaying method of Chinese medicine composition of the present invention is:
The preparation of reference substance solution: it is an amount of that the schisandrin reference substance decided in accurate title, adds methanol and process the solution that every 1ml contains 25 μ g, promptly gets.
The preparation of need testing solution: it is an amount of to get these article, and porphyrize is got about 1g, and accurate the title decides, and the accurate methanol 50ml that adds claims to decide weight; Supersound process (power 500W, frequency 40kHz) 1 hour is put coldly, claims to decide weight again, supplies the weight that subtracts mistake with methanol, shakes up; Filter, discard filtrating just, precision is measured subsequent filtrate 40ml, and evaporate to dryness, residue add dissolve with methanol and be transferred in the 10ml measuring bottle; Add methanol and be diluted to scale, shake up, filter, get subsequent filtrate, promptly get with microporous filter membrane (0.45 μ m).
Measure: according to accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of HPLC (appendix VI D), inject chromatograph of liquid, measure.
Chromatographic condition: with the octadecylsilane chemically bonded silica is filler; With methanol-water (53: 47) is mobile phase; The detection wavelength is 250nm; Number of theoretical plate calculates by the schisandrin peak and is not less than 3000.
Identify through above-mentioned thin layer liquid chromatograph, in the test sample chromatograph of Chinese medicine composition of the present invention, with control medicinal material and the corresponding position of reference substance chromatograph on, the speckle of apparent same color; During assay, every 1g contains Fructus Schisandrae Chinensis with schisandrin (C
24H
32O
7) meter, be no less than 0.15mg, preferably be no less than 0.20mg.
Chinese medicine composition of the present invention has the effect of nourishing the brain and improving intelligence, hypnotic, resisting fatigue, cold-resistant, anoxia enduring, enhancing humoral immunization, can be used for nourishing the brain and improving intelligence, tranquilizing by nourishing the heart, treatment brain overwork; Amnesia, the neurasthenia has a dizzy spell; The palpitation with fear insomnia, vexed easy to be tired, the fear of cold body is empty; Body loses pain in the lumbar region and senile dementia, looks patient's needs, but takes 1-10g every day.
The brain-strengthening of Chinese medicine composition of the present invention is effective, compares with existing brain-strengthening medicine, and its drug effect is better, and zoopery proves, brain-strengthening Chinese medicine composition of the present invention has the effect of hypnotic, resisting fatigue, cold-resistant, anoxia enduring, enhancing humoral immunization; The favorable reproducibility of the detection effect of quality determining method, highly sensitive.
Description of drawings
Fig. 1 is a Semen Ziziphi Spinosae cell photo, and wherein (a) is the palisade cells side; (b) be the palisade cells surface; (c) be the endotesta cell;
Fig. 2 is a Fructus Schisandrae Chinensis cell photo;
Fig. 3 is a Fructus Lycii cell photo;
Fig. 4 is the thin layer liquid chromatogram of Radix Angelicae Sinensis; Wherein 1. lack the Radix Angelicae Sinensis negative control; 2. embodiment 1 sample; 3. embodiment 2 samples; 4. embodiment 3 samples; 5. Radix Angelicae Sinensis control medicinal material;
Fig. 5 is the thin layer liquid chromatogram of Fructus Lycii, and wherein 1. lack the Fructus Lycii negative control; 2. embodiment 1 sample; 3. embodiment 2 samples; 4. embodiment 3 samples; 5. Fructus Lycii control medicinal material;
Fig. 6 is the thin layer liquid chromatogram of Herba Cistanches, wherein 1. misrun Herba Cistanches negative controls; 2. embodiment 1 sample; 3. embodiment 2 samples; 4. embodiment 3 samples; 5. echinacoside reference substance;
Fig. 7 is the thin layer liquid chromatogram of Fructus Schisandrae Chinensis, wherein 1. embodiment, 1 sample; 2. embodiment 2 samples; 3. embodiment 3 samples; 4. deoxyschizandrin reference substance; 5. schisandrin B reference substance; 6. schisandrin reference substance; 7. schisantherin A reference substance; 8. Fructus Schisandrae Sphenantherae control medicinal material; 9. Fructus Schisandrae Chinensis control medicinal material; 10. lack the Fructus Schisandrae Chinensis negative control;
Fig. 8 is the thin layer liquid chromatogram of Semen Ziziphi Spinosae, 1. acid deficient Semen Ziziphi Spinosae negative control; 2. embodiment 1 sample; 3. embodiment 2 samples; 4. embodiment 3 samples; 5. jujuboside A and jujuboside B reference substance;
Fig. 9 is the thin layer liquid chromatogram of Radix Ginseng; 1. the shortage of staff joins negative control; 2. embodiment 1 sample; 3. embodiment 2 samples; 4. embodiment 3 samples; 5. Radix Ginseng control medicinal material; 6. ginsenoside Rg1's reference substance.
The specific embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.
1, prescription:
Radix Angelicae Sinensis 33.3g, Concretio Silicea Bambusae 13.3g, Herba Cistanches (salt is processed) 26.7g, Dens Draconis (forging) 13.3g, Rhizoma Dioscoreae 26.7g, succinum 13.3g, Fructus Schisandrae Chinensis (wine steaming) 20g, Rhizoma Gastrodiae 6.7g, Semen Platycladi (stir-fry) 5.3g, Radix Salviae Miltiorrhizae 6.7g, Fructus Alpiniae Oxyphyllae (salt stir-fry) 20g, Radix Ginseng 6.7g, Radix Polygalae (liquorice beverage is processed) 13.3g, Flos Chrysanthemi 6.7g Rhizoma Anemones Altaicae 13.3g, Haematitum 10g, Arisaema Cum Bile 13.3g, Semen Ziziphi Spinosae (stir-fry) 53.3g, Fructus Lycii 26.7g.
Fructus Schisandrae Chinensis is the dry mature fruit of magnoliaceae schisandra in the prescription.Practise and claim " Radix Schisandrae Bicoloris ", but not " Fructus Schisandrae Sphenantherae "; Rhizoma Anemones Altaicae is concocted according to standard " Shandong Province's Chinese medicine processing standard 2002 (12 pages) " in the prescription.
2. method for making:
More than 19 the flavor, Haematitum, succinum, Concretio Silicea Bambusae are ground into fine powder respectively, ten Six-elements such as all the other Radix Angelicae Sinensis are ground into fine powder, with above-mentioned fine powder facing-up, sieve, mixing, the system granule, drying, encapsulated, process 1000, promptly get.
1, prescription:
Radix Angelicae Sinensis 25g; Concretio Silicea Bambusae 10g; Herba Cistanches (salt is processed) 20g; Dens Draconis (forging) 10g; Rhizoma Dioscoreae 20g; Succinum 10g; Fructus Schisandrae Chinensis (wine steaming) 15g; Rhizoma Gastrodiae 5g; Semen Platycladi (stir-fry) 4g; Radix Salviae Miltiorrhizae 5g; Fructus Alpiniae Oxyphyllae (salt stir-fry) 15g; Radix Ginseng 5g; Radix Polygalae (liquorice beverage is processed) 10g; Flos Chrysanthemi 5g; Rhizoma Anemones Altaicae 10g; Haematitum 7.5g; Arisaema Cum Bile 10g; Semen Ziziphi Spinosae (stir-fry) 40g; Fructus Lycii 20g.
Fructus Schisandrae Chinensis is the dry mature fruit of magnoliaceae schisandra in the prescription, practise and claim " Radix Schisandrae Bicoloris ", but not " Fructus Schisandrae Sphenantherae ".Rhizoma Anemones Altaicae is concocted according to standard " Shandong Province's Chinese medicine processing standard 2002 (12 pages) " in the prescription.
2, technology:
More than 19 the flavor, except that Haematitum, succinum, Concretio Silicea Bambusae list porphyrize powder, ten Six-elements such as all the other Radix Angelicae Sinensis are ground into fine powder; Sieve with Haematitum (50%), succinum, Concretio Silicea Bambusae fine powder, mixing adds an amount of sodium alginate mixing and uses water pill again; Or adopt the machine pill, polish drying.With Resina persicae, residue Haematitum coating, be drying to obtain.
1, prescription:
Radix Angelicae Sinensis 20g, Concretio Silicea Bambusae 10g, Herba Cistanches (salt is processed) 20g, Dens Draconis (forging) 10g, Rhizoma Dioscoreae 20g, succinum 15g, Fructus Schisandrae Chinensis (wine steaming) 15g, Rhizoma Gastrodiae 5g, Semen Platycladi (stir-fry) 4g, Radix Salviae Miltiorrhizae 5g, Fructus Alpiniae Oxyphyllae (salt stir-fry) 15g, Radix Ginseng 5g, Radix Polygalae (liquorice beverage is processed) 10g, Flos Chrysanthemi 5g Rhizoma Anemones Altaicae 10g, Haematitum 7g, Arisaema Cum Bile 10g, Semen Ziziphi Spinosae (stir-fry) 40g, Fructus Lycii 20g.
Fructus Schisandrae Chinensis is the dry mature fruit of magnoliaceae schisandra in the prescription.Practise and claim " Radix Schisandrae Bicoloris ", but not " Fructus Schisandrae Sphenantherae "; Rhizoma Anemones Altaicae is concocted according to standard " Shandong Province's Chinese medicine processing standard 2002 (12 pages) " in the prescription.
2. method for making:
More than 19 the flavor, Haematitum, succinum, Concretio Silicea Bambusae are ground into fine powder respectively, ten Six-elements such as all the other Radix Angelicae Sinensis are ground into fine powder, with above-mentioned fine powder facing-up, sieve, mixing, the system granule, drying, encapsulated, process 1000, promptly get.
1, prescription:
Radix Angelicae Sinensis 35g, Concretio Silicea Bambusae 15g, Herba Cistanches (salt is processed) 27g, Dens Draconis (forging) 12g, Rhizoma Dioscoreae 24g, succinum 12g, Fructus Schisandrae Chinensis (wine steaming) 18g, Rhizoma Gastrodiae 8g, Semen Platycladi (stir-fry) 5.5g, Radix Salviae Miltiorrhizae 8g, Fructus Alpiniae Oxyphyllae (salt stir-fry) 21g, Radix Ginseng 8g, Radix Polygalae (liquorice beverage is processed) 15g, Flos Chrysanthemi 8g Rhizoma Anemones Altaicae 15g, Haematitum 8g, Arisaema Cum Bile 15g, Semen Ziziphi Spinosae (stir-fry) 50g, Fructus Lycii 25g.
Fructus Schisandrae Chinensis is the dry mature fruit of magnoliaceae schisandra in the prescription.Practise and claim " Radix Schisandrae Bicoloris ", but not " Fructus Schisandrae Sphenantherae "; Rhizoma Anemones Altaicae is concocted according to standard " Shandong Province's Chinese medicine processing standard 2002 (12 pages) " in the prescription.
2. method for making:
More than 19 the flavor, Haematitum, succinum, Concretio Silicea Bambusae are ground into fine powder respectively, ten Six-elements such as all the other Radix Angelicae Sinensis are ground into fine powder, with above-mentioned fine powder facing-up, sieve, mixing, the system granule, drying, encapsulated, process 1000, promptly get.
The test of pesticide effectiveness of embodiment 5 Chinese medicinal composition capsules
[test material]
Receive the Chinese medicinal composition capsules of reagent thing: embodiment 1 preparation: chocolate brown powder, every capsules contain crude drug powder powder 0.3g crude drug, are made into required concentration with preceding with distilled water.
The dosage design: according to the clinical consumption 1.8g crude drug of people/sky, calculate by " pressing the dose,equivalent ratio of body surface area conversion between humans and animals ", mice is a low dosage with 0.3g crude drug/kg, is equivalent to clinical equivalent dosage; Middle dosage is 2 times of low dosage, and high dose is 1.5 times of middle dosage.Cavia porcellus is a low dosage with 0.2g crude drug/kg, is equivalent to clinical equivalent dosage, and middle dosage is 2 times of low dosage, and high dose is 1.5 times of middle dosage.
Positive control drug: pentobarbital sodium, China Drug Co.'s Beijing purchasing and supply station import packing, lot number 01062222; Radix Ginseng panax gineeng purchases the pharmacy of Tongrentang in Qingdao; Propranolol, Shanghai City the 15 pharmaceutical factory of pharmaceuticals produces, lot number 010908; Levamisole hydrochloride, Junan, Shandong Province pharmaceutical factory produces, lot number 010602
Animal subject: Kunming mouse 19-21g, totally 340, wherein male 204, available from Qingdao of Shandong province laboratory animal and zoopery center, the quality certification number: Shandong kinoplaszm word 200002001.Cavia porcellus 170-200g, totally 3, male and female half and half, available from Institute of Experimental Animals, Chinese Academy of Medical Sciences's breeding field, the quality certification number: the capital is moving is betrothed to 017.
Reagent: anhydrous glucose, Shanghai chemical reagent purchasing and supply station chemical reagent work produces, lot number 010320; Sodium citrate .2H
2O produces 010414 by chemical industry station, Qingdao chemical reagent work, lot number; 20010705; Citric acid is produced lot number 010314 by chemical industry station, Qingdao chemical reagent work; Sodium chloride is produced lot number 010411 by Beijing chemical reagents corporation; Normal saline is produced lot number 010522 by Hangzhou animal drug factory; Sodica calx, analytical pure, west, Shantou City Long Huagongchang, lot number 0106201; Pentobarbital sodium, China Drug Co.'s Beijing purchasing and supply station import packing, lot number 01062222
The glass jar of instrument: 50cm * 30cm * 25cm, aluminium flake (2-3g), thermometer, stopwatch; The 250ml wide mouthed bottle; Refrigerator; Germany produces Sartorius electronic balance, model: BP110S; Pressure cooker, refrigerator, conical flask, 721-ultraviolet spectrophotometer (a day island proper Tianjin produces), liquid-transfering gun, test tube.
[test method and result]
Chinese medicinal composition capsules to pentobarbital sodium sub-threshold lull dosage influence
Test method: get 40 of healthy Kunming white mice, body weight 18～20g, male and female dual-purpose.Experimental animal is divided into 4 groups at random by body weight and sex: (1) matched group, n=10; (2) Chinese medicinal composition capsules group, 0.3g/kg, n=10; (3) Chinese medicinal composition capsules group 0.6g/kg, n=10; (4) Chinese medicinal composition capsules group, 0.9g/kg, n=10 is made into desired concn with preceding with distilled water.
Each administration group is according to dosage irritated stomach every day to Chinese medicinal composition capsules once, continuous seven days.Matched group is irritated stomach and is given distilled water.After the last administration 1 hour, by the pentobarbital sodium of lumbar injection subliminal hypnosis dosage.Before experiment, do the preliminary experiment of pentobarbital sodium sub-threshold lull dosage with same batch of white mice of laboratory animal, subliminal hypnosis dosage is 32mg/kg.Observing in 30 minutes behind the injection pentobarbital sodium has how many animals areflexia anyway, anyway areflexia reaches the animal of animal for sleeping more than 1 minute.Matched group is lumbar injection pentobarbital sodium 32mg/kg only, compares the sleeping percentage rate of administration group and mice in control group, with 2 groups of significance test of x.
Experiment is carried out under 24 ℃ of-25 ℃ of quiet environment.
Anyway the standard of areflexia is: animal back is placed on the black sheet iron down gently, and iron sheet keeps more than 30 ℃, and vibration gently, and animal back posture down keeps person more than 30 seconds.
The result sees table 1, and the result shows: Chinese medicinal composition capsules and pentobarbital sodium have synergism, can significantly increase the sleeping percentage rate of the inductive mice of pentobarbital sodium sub-threshold dose (with the matched group comparison,
△P＜0.05,
△ △P＜0.01), thus show that Chinese medicinal composition capsules has the effect of hypnotic.
Table 1 Chinese medicinal composition capsules is to the sleeping percentile influence of pentobarbital sodium mice of injection subliminal hypnosis dosage
| Group | Example number (n) | Dosage | Sleeping percentage rate (%) |
| Matched group Chinese medicinal |
10 10 10 10 | -- 0.3g/kg 0.6g/kg 0.9g/kg | 0 30 60 △ 90 △△ |
Annotate: compare with matched group,
△P＜0.05,
△ △P＜0.01.
2. Chinese medicinal composition capsules is to the antifatigue effect of mice
Test method: get 50 of healthy Kunming white mice, body weight 18～20g, male.Experimental animal is weighed respectively, and numbering is divided equally 5 groups by body weight: (1) matched group, normal saline, n=10; (2) Radix Ginseng, 0.4g/kg, n=10; (3) Chinese medicinal composition capsules group, 0.3g/kg, n=10; (4) Chinese medicinal composition capsules group, 0.6g/kg, n=10; (5) Chinese medicinal composition capsules group, 0.9g/kg, n=10 is made into desired concn with preceding with distilled water.
The administration group is according to dosage irritated stomach every day to Chinese medicinal composition capsules once; Continuous seven days. fasting is 12 hours before the last administration; The last administration experimentized after 30 minutes. before after the last administration, experimentizing; To claim the mice body weight, and give each mouse tail root bundle one aluminium flake by body weight 10%. it is that (27 ± 0.5) ℃, the depth of water are in the glass jar of 25cm that the mice that bears a heavy burden is dropped into water temperature, lets its swimming; Clocking immediately. matched group is irritated stomach to normal saline every day; The positive drug group is irritated stomach every day and is given Radix Ginseng. and can not swim out of the water surface 10 seconds with the mouse head submerged serves as that muscle power is exhausted, the observed and recorded mice was hauled oneself willingly in the entry to the time that muscle power is exhausted, was swimming time. and the result organizes a significance test with the t check.
Result of the test is seen table 2; The result shows: middle and high two dose groups of Chinese medicinal composition capsules are ability significant prolongation mice swimming time all; Average swimming time all relatively has significant differences (P＜0.01) than matched group, but low dose group and matched group compare there was no significant difference.The antifatigue effect that the presentation of results Chinese medicinal composition capsules has.
Table 2 Chinese medicinal composition capsules is to the antifatigue effect of mice
| Group | Example number (n) | Dosage (g/kg) | Swimming time (min) |
| Matched group Radix Ginseng group Chinese medicinal |
10 10 10 10 10 | --- 0.4 0.3 0.6 0.9 | 6.10±1.48 11.7±1.64 △△ 6.2±1.47 9.5±1.67 △△ 9.5±1.55 △△ |
Annotate: compare with matched group,
△P＜0.05,
△ △P＜0.01.
3. Chinese medicinal composition capsules is to the cold-resistant function of mice
Test method: get 50 of healthy Kunming white mice, body weight 18～20g, male and female half and half.Test mice is weighed respectively, and numbering is divided equally 5 groups by the body weight sex: (1) matched group, distilled water, n=10; (2) Radix Ginseng, 0.4g/kg.(3) Chinese medicinal composition capsules group, 0.3g/kg, n=10; (4) Chinese medicinal composition capsules group, 0.6g/kg, n=10; (5) Chinese medicinal composition capsules group, 0.9g/kg, n=10 is made into desired concn with preceding with distilled water.
The administration group is according to dosage irritated stomach every day to Chinese medicinal composition capsules once, continuous seven days. after the last administration, clock immediately. and matched group is irritated stomach to distilled water every day; The positive drug group is irritated stomach every day and is given Radix Ginseng. and the last administration is after 30 minutes, mice is positioned in the refrigerator of (5 ± 1) ℃; Every cage is put a mice, after clocking .2 hour mice is taken out the survival of record mice; Calculate the percentage survival of each group, organize a significance test with x 2 checks.
Result of the test is seen table 3, and the result shows: Chinese medicinal composition capsules 0.9g/kg dose groups can improve the tolerance to cold of mice in cold environment, and percentage survival and matched group relatively have significant difference (P＜0.05); Chinese medicinal composition capsules 0.6g/kg dose groups and 0.3g/kg dose groups and matched group be there was no significant difference relatively.The result shows that the Chinese medicinal composition capsules heavy dose has been taken for a long time and sets upright the effect of dispeling cold.
Table 3: Chinese medicinal composition capsules is to the cold-resistant function of mice
| Group | Example number (n) | Dosage (g/kg) | Percentage survival (%) |
| Matched group Radix Ginseng Chinese medicinal |
10 10 10 10 10 | --- 0.4 0.3 0.6 0.9 | 30 100 △△ 60 80 90 △ |
Annotate: compare with matched group,
△P＜0.05,
△ △P＜0.01
4. Chinese medicinal composition capsules is to the influence of mice normal pressure anoxia enduring
Test method: get 50 of healthy Kunming white mice, body weight 18～20g, male and female half and half.Experimental animal is weighed respectively, and numbering is divided equally 5 groups by the body weight sex: (1) matched group, normal saline, n=10; (2) positive drug group, propranolol, 20mg/kg, n=10 (3) Chinese medicinal composition capsules group, 0.3g/kg, n=10; (4) Chinese medicinal composition capsules group, 0.6g/kg, n=10; (5) Chinese medicinal composition capsules group, 0.9g/kg, n=10
The administration group is according to dosage irritated stomach every day to Chinese medicinal composition capsules once, after the administration in continuous seven days, the 8th day, clocks immediately; Matched group is irritated stomach to normal saline every day, and the positive drug group is irritated stomach to propranolol every day, and administration is after 30 minutes; Mice is positioned in the 250ml wide mouthed bottle that fills the 15g sodica calx, one every bottle, tight with vaseline applicating bottle flap; Make it air tight; With respiratory arrest is dead index, and the record mice is organized a significance test from enclosing the time (time-to-live) extremely dead in the bottle with the t check.
Result of the test is seen table 4, and the result shows: high, medium and low three dose groups of Chinese medicinal composition capsules all can significantly improve mice normal pressure hypoxia-bearing capability, and mean survival time is all than matched group significant prolongation (P＜0.05, P＜0.01); Dose groups and Chinese medicinal composition capsules low dose group relatively have significant difference (P＜0.05) in the Chinese medicinal composition capsules; Dose groups relatively has significant difference (P＜0.01) in Chinese medicinal composition capsules high dose group and the Chinese medicinal composition capsules.Presentation of results, Chinese medicinal composition capsules can improve mice normal pressure hypoxia-bearing capability, and present tangible dose-effect relationship between each dose groups.
Table 4: Chinese medicinal composition capsules is to the influence of mice normal pressure anoxia enduring
| Group | Example number (n) | Dosage (g/kg) | Time-to-live (min) |
| Matched group propranolol Chinese medicinal |
10 10 10 10 10 | --- 0.02 0.3 0.6 0.9 | 21.75±0.96 34.46±1.51 △△ 23.41±1.47 △ 28.99±1.80 △△ 31.20±1.90 △△ |
Annotate: compare with matched group,
△P＜0.05,
△ △P＜0.01.
5. Chinese medicinal composition capsules is to the influence of mice serum hemolysin
Test method:
Experimental animal divides into groups: get 50 of healthy Kunming white mice, body weight 19～21g, male and female half and half.Cavia porcellus 170-200g, totally 3, male and female dual-purpose, healthy tame chicken.Will, 50 mices are divided 5 groups at random by body weight, sex, i.e. matched group, levamisole group, Chinese medicinal composition capsules group (0.3g/kg, 0.6g/kg, 0.9g/kg)
The preparation of Alsever ' s liquid: citric acid trisodium 2H2O 8.0g, citric acid 0.5g, anhydrous glucose 18.7g, sodium chloride 4.2g, distilled water adds to 1000ml, filters packing, 10 pounds of high pressure 20min (4 ℃ of refrigerators are preserved subsequent use)
The preparation of chicken erythrocyte suspension: the sterile working, from venous blood collection, put the 100ml conical flask, add Alsever ' the s liquid that is equivalent to 5 times of Sanguis Gallus domesticus volumes; Mixing is put in 4 ℃ of freezer storages. and face with preceding and give a baby a bath on the third day after its birth time preceding 2 centrifugal 1500r/min with normal saline; Each 5min abandons the leukocytic cream at supernatant and interface, at last continuous 2 centrifugal 2000r/min; Each 5min, constant to the packed cell volume value, be made into 5% suspension.
The complement preparation: 3 Cavia porcelluss are drawn blood from heart, and separation of serum mixes, and is made into 10% (1: 10) with normal saline.
The mensuration of mice serum hemolysin: every Mus lumbar injection 5% normal saline chicken red blood cell suspension 0.2ml carries out immunity, gastric infusion simultaneously, and successive administration 7 days after the last administration 30 minutes, is plucked eyeball and is got blood, centrifugalize serum.Get serum, with 100 times of normal saline dilutions.Get dilute serum 1ml, with 5% chicken erythrocyte suspension 0.5ml, 10% complement 0.5ml mixes; After being incubated 30min in 37 ℃ of calorstats, stopped reaction in 0 ℃ of refrigerator, centrifugal; Get supernatant in the 540nm place colorimetric; Measure optical density (OD), other establishes the not blank of increase serum, transfers the benchmark of " 0 " when getting its supernatant as colorimetric.As the index of judging serum hemolysin, carry out the t check, comparable group differences property with optical density (OD) reading.
Experimental result is seen table 5.The result shows: compare with matched group; Middle and high two dose groups of Chinese medicinal composition capsules all can increase the quantity of mice serum hemolytic antibody; The generation of enhancing antibody; Average OD value through statistical procedures matched group and middle and high two dose groups of Chinese medicinal composition capsules all has significant differences (p＜0.01), but Chinese medicinal composition capsules low dose group and matched group compare not statistically significant.The result shows that Chinese medicinal composition capsules has the effect of the humoral immunization that strengthens mice.
Table 5: Chinese medicinal composition capsules is to the influence of mice serum hemolysin (X ± SD)
Annotate: compare with matched group,
*P＜0.05,
*P＜0.01
Brief summary:
Chinese medicinal composition capsules is made up of 19 flavor Chinese medicines such as Radix Angelicae Sinensis, Fructus Lycii, Semen Ziziphi Spinosae; Have nourishing the brain and improving intelligence, the effects such as mending body of sleeping peacefully, be used for brain overwork, amnesia, neurasthenia, the insomnia of having a dizzy spell, palpitate with fear, vexedly be prone to tired, fear of cold body void, body loses diseases such as lumbago and senile dementia.Existing cure mainly according to its function, Chinese medicinal composition capsules has been carried out it influences test, anti-fatigue test, low temperature resistant test, hypoxia endurance test and test for humoral immunity to sleep.Result of the test shows, Chinese medicinal composition capsules has effects such as tangible hypnotic, resisting fatigue, cold-resistant, anoxia enduring, enhancing body fluid, and it provides the pharmacodynamic experiment foundation for clinical practice.
The nourishing the brain and improving intelligence mechanism of embodiment 6 Chinese medicine composition pills
Research thinks that choline can system play facilitation in study, memory, so the neural maincenter of concentrating of choline ability is impaired or use anticholinergic agents that study, memory ability are descended.Hyoscine is injected the ventricles of the brain or Hippocampus can make memory keep weakening, pace of learning slows down, and it can disturb obtaining of memory, and can cause retrograde amnesia.It is closely similar with normal old people's amnesia that healthy young people is taken the dysmnesia that hyoscine causes, old people's amnesia is likely then that cholinergic systemic-function in the brain is weak and causes.
Hippocampus is considered to relevant with recent memory, damages the bilateral Hippocampus and can make rat diving tower avoidance response be difficult to consolidate, the recent memory forfeiture.Hippocampus one edge system has abundant cholinergic fiber and cholinoceptive cell and receptor, and septal area-Hippocampus-limbic system is one of path of its effect.Therefore adopt the brain-strengthening mechanism of following experiment discussion Chinese medicine composition of the present invention:
One, experiment material:
91 of healthy SD bull rat, body weight 180-220g is provided by animal housing of Shandong Medical University, raises drink tap water, the standard feed that food animal housing provides under the room temperature; Adopt the pill of embodiment 2 preparations, the unconcerned sour hyoscine of hydrogen is produced by Guangdong emigrant's light pharmaceutical factory.Key instrument has MG-2 type labyrinth, remittance gulf l type G Stereotaxic apparatus, and Switzerland produces AVL one 945 blood gas analyzers.
Two, research index:
Obtain and reproduce the obstacle index: select the reaction times before four index 1 maze learnings reach standard for use, during the overall reaction motion, initiatively rate of avoidance, correct response rate.
Septal area, Hippocampus breaking test are selected the preceding index of reaction times of the standard of reaching for use.
The influence of brain blood oxygen is selected for use three indexs: blood oxygen pressure, oxygen content and blood oxygen saturation.
Three, experiment is divided into groups
1, to remembering the influence of the acquisition and the process of reproduction: 50 of rats, be divided into 5 groups at random, irritate stomach (being equivalent to clinical dosage) with 0.9% normal saline or 0.75% Chinese medicine composition suspension respectively; 0.1ml/10g/ it is inferior; 3 times/day, the labyrinth test is carried out in row labyrinth training after continuous 5 days behind the 24h.See table 6.
Table 6 Chinese medicine composition pill is to dividing into groups from the experiment that the Mus memory obtains and reproduction influences greatly
2, the district is accompanied in two surveys, Hippocampus is destroyed the influence of rat memory: 25 of rats are divided into 5 groups at random; Carry out labyrinth training respectively, undergo surgery after reaching standard (continuous 10 secondary responses 9 times are correct, other with); Treat to irritate stomach 3 times/day with 0.9% normal saline or 0.75% Chinese medicine composition pill suspension after animal regains consciousness 0.1ml/10g/ time (septal area destruction group adopts the method for the administration of drinking water, and dosage is suitable); After continuous 7 days, carry out the labyrinth test more respectively.Observe its memory state.
3, the Chinese medicine composition pill is to the influence of brain blood oxygen: 16 of SD rats, be divided into four groups at random, and irritate stomach with 0.9%NS and 0.75%, 7.5%, 37.5% Chinese medicine composition pill suspension respectively; 3 times/day; 0.1ml/IOg/ inferior, after continuous 20 days, carotid artery is got blood and is carried out blood gas analysis.
Four, result of study
1, the Chinese medicine composition pill is to remembering the influence of the acquisition and the process of reproduction: the Chinese medicine composition pill can make the preceding reaction times of the standard that reaches of memory acquisition disturbance rat reduce the accuracy raising.Not remarkable to reproducing the effect of obstacle rat, make the active rate of avoidance decline that obtains and reproduce the obstacle rat.See table 7.
The influence that table 7 Chinese medicine composition pill obtains and reproduces memory
Explain: check with variance analysis when reaction times and overall reaction move before up to standard: initiatively rate of avoidance and correct response rate are used X
2Check α=0.05
*Compare P＜0.05 △ with the blank group and compare P＜0.05 with the hyoscine group
*Compare P＜0.01 △ △ with the blank group and compare P＜0.01 with the hyoscine group
2, the Chinese medicine composition pill is to two surveys septal area; Hippocampus is destroyed the influence of the memory of rat: the destruction of septal area and Hippocampus has all successfully been set up the model of memory impairment; The reaction times that septal area and Hippocampus destruction group reaches before the study standard during test all is higher than the blank group, uses the effect of then having played this infringement of antagonism behind the Chinese medicine composition pill Chinese medicine composition pill.See table 8.
Table 8 Chinese medicine composition pill destroys the influence of rat memory to septal area, Hippocampus
Explain: statistical method is the t check, α=0.05
*The blank group is than P＜0.01,
△ △ organizes than P＜0.01 with Hippocampus destruction than P＜0.01 with the blank group.
3, the Chinese medicine composition pill is to the influence of brain blood oxygen: have only 0.75% Chinese medicine composition pill group to compare blood oxygen pressure and blood oxygen saturation with the blank group and improve, oxygen content is not had influence.See table 4.
Obviously, the memory acquisition disturbance that the Chinese medicine composition pill can the antagonism hyoscine causes makes the bilateral septal area, and the memory that Hippocampus is destroyed rat improves, and explain that the Chinese medicine composition pill promotes to learn, memory impairments maybe be relevant with cholinergic system; And the Chinese medicine composition pill can improve blood oxygen pressure and blood oxygen saturation.
The experimentation of embodiment 7 Chinese medicine composition pills to influencing from Mus study, memory behavior greatly
One, test material: 30 of healthy SD bull rat, body weight 180-220g is provided by animal housing of Shandong Medical University, raises drink tap water, food standard feed under the room temperature.
The Chinese medicine composition pill of embodiment 2 preparations; Piracetam (2-Pyrrolidone acetamide) is that Weihai Xinhua pharmaceutical factory produces, and key instrument is MG-2 type trisection radiant type labyrinth.
Two, experimental technique: 30 SD rats are divided into three groups at random, 10 every group.
1, blank group: irritate stomach, 3 times/day 0.1ml/10g/ time with 0.9% normal saline.
2, piracetam group (experiment contrast group): irritate stomach, 3 times/day 0.03ml/10g/ time with 10% piracetam solution.
3, contain Chinese medicine composition pill (experimental group): with 0.75% ' contain Chinese medicine composition pill suspension to irritate stomach, 3 times/day 0.1ml/10g/ time.
The consumption of Chinese medicine composition pill and piracetam is the clinical consumption according to the people, according to the body surface area conversion of people and rat.
Above-mentioned three groups of rats carry out the learning training in labyrinth before respectively at medication.Experiment is quietly being carried out in the environment, during experiment rat is put into the labyrinth and adapts to 5 minutes earlier, begins experiment then, and the support arm that light is arranged is the place of safety no power, and the place of safety is with random order conversion, and energising voltage is 30-50V.Directly escape to the place of safety person after rat is shocked by electricity and be " correct response ", otherwise belong to " wrong reaction ".There are 9 correct responses promptly to calculate the standard that reaches association continuous 10 times.
Three, observation index:
1, reach the number of times of required reaction before association's standard: desired times is few more, and learning and memory is good more;
2, overall reaction when motion: refer to each secondary response of accumulative total, escape the summation of required time till the bright place of safety of lamp to rat from bright beginning of signal lights;
3, active avoidance response rate: rat was accomplished the percentage rate that the number of times that reacts accounts for the overall reaction number of times in 5 seconds.
4, correct response rate: refer to that " correct response " number of times accounts for the percentage rate of overall reaction number of times.
Four, experimental result:
After the medication, experimental group is compared with the blank group with the piracetam group, and the preceding reaction times of the standard that reaches reduces, and the correct response rate improves, and experimental group active rate of avoidance is higher; There is not significant difference when overall reaction moves between each group.See table 9-12 for details.
Table 9:-reaches required reaction times (unit: inferior) before the standard
During the motion of table 10 overall reaction unit: second
Table 11:-active avoidance response rate (%)
Table 12 correct response rate (%)
Explain: statistical method table 1, table 2 are variance analysis; Table 3, table 4 are t check, α=0.05
*For with the blank group than P＜0.05;
*For with the blank group than P＜0.01; △ △ and blank group are than P＜0.01.
Piracetam is the medicine of known enhancement memory, and this experiment finds that the Chinese medicine composition pill can make rat maze learning number of times and correct response rate improve, and explains that this medicine has the study of the rat of promotion, memory impairments.
The clinical trial of embodiment 8 Chinese medicine compositions
By Zhongxin Hospital, Qingdao, Qingdao Eighth People's Hospital, 3 tame hospitals of Shibei District, Qingdao City hospital carried out this Chinese medicinal composition capsules treatment Cure for insomnia hyperactivity of fire caused by deficiency of YIN, syndrome of deficiency of both heart and spleen has carried out clinical trial.Final report is following.
One, test objective
This clinical trial is treated the front and back sleep quality through observing, memory, and expression, attention is had a dizzy spell, the situation of change of soreness of the waist and knees tongue arteries and veins, the clinical efficacy of evaluation Chinese medicinal composition capsules Cure for insomnia card.
Two, test method
(1) EXPERIMENTAL DESIGN type, principle
1, EXPERIMENTAL DESIGN type: parallel-group control design at random.
2, contrast: the contrast medicine is selected ANSHEN BUXIN WAN for use.
3, blind method design: adopt single blind method.
(2) case is selected
1, diagnostic criteria
Insomnia's Western medicine diagnose standard (" Chinese psychiatric department association's mental sickness classification schemes and diagnostic criteria ");
Tcm diagnosis standard: insomnia;
(1) classical symptom of insomnia is arranged: difficulty falling asleep, awakening often, sleep and unstable or wake up after can not sleep again; Wake up morning too early; Can not fall asleep night, daytime lethargic and drowsy state; Do not have enough sleep 5 hours.
(2) history of repeated attack is arranged.
(3) Clinical typing
Slightly: sleep is awakened often or is slept and unstable, and it is too early to wake up morning, but does not influence work.
Moderate: do not have enough sleep 4 hours, but still can adhere to operate as normal.
Severe: lie awake all night, be difficult to adhere to operate as normal.
(3) therapeutic scheme
1. test medication title and specification
The Chinese medicinal composition capsules of test medication: embodiment 1 preparation, specification: every 0.3g, lot number 030501.
Contrast medicine: ANSHEN BUXIN WAN, Yantai, the Shandong pharmaceutical factory of traditional Chinese medicine.Specification: 2g/ ball, lot number 030404.
(4) observation index
1. main curative effect index
Primary symptom: the keeping the score of insomnia.
Three, result of the test
(1) observation index result
1. primary symptom
Difference no statistical significance between the group that the insomnia baseline is kept the score has comparability between two groups.Treat the equal not statistically significant of difference between the group of the 14th day, the 28th day drop-out value of keeping the score and keep the score.Treat the difference of keeping the score in the 14th day, the 28th day for two groups the height statistical significance is all arranged with the baseline comparison.See table 14,15.
Table 13: two groups of situations of change (PPS) of keeping the score of having a sleepless night
Table 14: two groups of situations of change (FAS) of keeping the score of having a sleepless night
2. inferior disease
Difference no statistical significance between the group that the hypomnesis baseline is kept the score has comparability between two groups.Treat the equal not statistically significant of difference between the group of the 14th day, the 28th day drop-out value of keeping the score and keep the score.Treat the difference of keeping the score in the 14th day, the 28th day for two groups the height statistical significance is all arranged with the baseline comparison.
Difference no statistical significance between the group that absent minded baseline is kept the score has comparability between two groups.Treat the equal not statistically significant of difference between the group of the 14th day, the 28th day drop-out value of keeping the score and keep the score.Treat the difference of keeping the score in the 14th day, the 28th day for two groups the height statistical significance is all arranged with the baseline comparison.
Difference no statistical significance between the group that the physical fatigue and lassitude of spirit baseline is kept the score has comparability between two groups.Treat the equal not statistically significant of difference between the group of the 14th day, the 28th day drop-out value of keeping the score and keep the score.Treat the difference of keeping the score in the 14th day, the 28th day for two groups the height statistical significance is all arranged with the baseline comparison.
Difference no statistical significance between the group that the baseline of having a dizzy spell is kept the score has comparability between two groups.Treat the equal not statistically significant of difference between the group of the 14th day, the 28th day drop-out value of keeping the score and keep the score.Treat the difference of keeping the score in the 14th day, the 28th day for two groups the height statistical significance is all arranged with the baseline comparison.
Difference no statistical significance between the group that the soreness of the waist and knees baseline is kept the score has comparability between two groups.Treat the equal not statistically significant of difference between the group of the 14th day, the 28th day drop-out value of keeping the score and keep the score.Treat the difference of keeping the score in the 14th day, the 28th day for two groups the height statistical significance is all arranged with the baseline comparison.
Difference has statistical significance between the group of the 14th day dizzy dizzy disappearance rate.
3. symptom and sign integration
Difference no statistical significance between the group before the total mark treatment has comparability between two groups.Treat the equal not statistically significant of difference between the group of the 14th day, the 28th day total mark drop-out value.Two groups of difference of treating the 14th day, the 28th day total mark and baseline comparison all have the height statistical significance.
4. comprehensive therapeutic effect analysis
The difference no statistical significance that meets two groups of comprehensive therapeutic effects of scheme set, total effective rate, the difference not statistically significant of cure rate.The 95%CI that total effective rate differs (7.04,18.99).
The difference no statistical significance of two groups of comprehensive therapeutic effects of complete analysis collection, total effective rate, the difference of cure rate has statistical significance.The 95%CI that total effective rate differs (6.58,19.57).
For getting rid of center effect, with the difference not statistically significant of two groups of total effective rates of center layering, the difference between each center is not statistically significant also.
Four, sum up
1. effectiveness
Treat the 14th day dizzy dizzy disappearance rate test group and be superior to matched group.
Meet scheme set test group total effective rate 60.38%, matched group total effective rate 66.36%.Complete analysis collection test group total effective rate 59.25%, matched group total effective rate 65.74%.The difference not statistically significant of two groups of comprehensive therapeutic effects.
2. safety
The difference not statistically significant of two groups of safety evaluatios.
The quality testing of embodiment 9 Chinese medicinal composition capsules
[assay] measured according to HPLC (appendix VI D).
Chromatographic condition and system suitability test: with the octadecylsilane chemically bonded silica is filler; With methanol-water (53: 47) is mobile phase; The detection wavelength is 250nm.Number of theoretical plate calculates by the schisandrin peak should be not less than 3000.
The preparation of reference substance solution: it is an amount of that the schisandrin reference substance decided in accurate title, adds methanol and process the solution that every 1ml contains 25 μ g, promptly gets.
The preparation of need testing solution: the sample of getting embodiment 1-4 preparation is an amount of, and porphyrize is got about 1g, and accurate the title decides, the accurate methanol 50ml that adds; Claim decide weight, supersound process (power 500W, frequency 40kHz) 1 hour is put coldly, and weight decided in title again; Supply the weight that subtracts mistake with methanol, shake up, filter, discard filtrating just; Precision is measured subsequent filtrate 40ml, and evaporate to dryness, residue add dissolve with methanol and be transferred in the 10ml measuring bottle, add methanol and are diluted to scale; Shake up, filter, get subsequent filtrate, promptly get with microporous filter membrane (0.45 μ m).
Measure: accurate respectively reference substance solution and each 10 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly get.
Table 15 sample determination result
[evaluation of thin layer liquid chromatograph]
(1) get these article, put microscopically and observe: plant skin palisade cells brownish red, norma lateralis is the string palisade to be arranged, and polygon is seen on the surface, and cell is big slightly, includes the dark-brown thing; The pale brown color of endotesta cell, surface see rectangle or class is square, and the anticlinal wall beaded thickens (Semen Ziziphi Spinosae), sees Fig. 1.Plant the pale brown color of skin epidermis stone cell, a type polygon is seen on the surface, and wall is thicker, and the hole ditch is fine and closely woven, and cell contains burgundy thing (Fructus Schisandrae Chinensis), sees Fig. 2.Plant skin stone cell surface and see irregular polygon, wall thickness, wavy bending, laminated striation clear (Fructus Lycii) is seen Fig. 3.
(2) get each 4g of sample of embodiment 1,2,3, porphyrize adds ethanol 40ml, and supersound process 20 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution.Other gets Radix Angelicae Sinensis control medicinal material 1g, adds ethanol 10ml, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VI B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with cyclohexane extraction-ethyl acetate (9: 1), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.Collection of illustrative plates is seen Fig. 4.
(3) get the sample 5g of embodiment 1,2,3, porphyrize adds water 100ml, and heated and boiled 15 minutes is put cold; Centrifugal, get supernatant, add ethyl acetate extraction 3 times, each 50ml; Combined ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate 2ml makes dissolving, as need testing solution.Other gets Fructus Lycii control medicinal material 1g, shines medical material solution in pairs with legal system.According to thin layer chromatography (appendix VI B) test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with toluene-ethyl acetate-formic acid (6: 4: 0.2), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.Collection of illustrative plates is seen Fig. 5.
(4) get the sample 10g of embodiment 1,2,3, porphyrize, the 60ml that adds diethyl ether, supersound process 30 minutes is put cold; Filter, discard ether solution, residue volatilizes solvent, adds methanol 100ml, supersound process 30 minutes; Put coldly, filter, the filtrating evaporate to dryness, residue adds water 10ml slight fever makes dissolving, through D101 macroporous adsorptive resins (internal diameter 1.5cm; Long 15cm), water 70ml eluting discards eluent, and reuse 20% ethanol 70ml eluting discards eluent; Continue with 60% methanol 100ml eluting, collect eluent, evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution.Other gets the echinacoside reference substance, adds methanol and processes the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography (appendix VI B) test, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, be developing solvent with methanol-acetic acid-water (2: 1: 7), launch, take out, dry, put under the ultra-violet lamp (365nm) and inspect.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the fluorescence speckle of same color.Collection of illustrative plates is seen Fig. 6.
(5) get Fructus Schisandrae Chinensis control medicinal material 1g, add ethanol 10ml, supersound process 20 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution.Get schisandrin B reference substance, schisandrin reference substance again, add methanol respectively and process the solution that every 1ml contains 1mg, as reference substance solution.Test according to thin layer chromatography (appendix VI B); Drawing need testing solution 10 μ l and above-mentioned control medicinal material solution and each 5 μ l of reference substance solution under the item of [discriminating] (2), put respectively on same silica GF254 lamellae, is developing solvent with cyclohexane extraction-ethyl acetate (7: 2.5); Launch; Take out, dry, put under the ultra-violet lamp (254nm) and inspect.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.Collection of illustrative plates is seen Fig. 7.
(6) get the need testing solution under the item of [discriminating] (4), evaporate to dryness, residue add water 15ml makes dissolving, extracts 3 times with water saturated n-butyl alcohol jolting; Each 15ml merges n-butyl alcohol liquid, with ammonia solution washing 2 times, and each 15ml; N-butyl alcohol liquid evaporate to dryness, residue add methanol 1ml makes dissolving, as need testing solution.Other gets jujuboside A and jujuboside B reference substance, adds methanol and processes the mixed solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography (appendix VI B) test, draw above-mentioned need testing solution 10 μ l and reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate; With water saturated n-butyl alcohol is developing solvent, launches, and takes out; Dry, spray is inspected with 2% vanillin sulfuric acid solution immediately.In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color.Collection of illustrative plates is seen Fig. 8.
(7) get Radix Ginseng control medicinal material 1g, add methanol 50ml, supersound process 30 minutes is put coldly, filters; The filtrating evaporate to dryness, residue adds water 10ml slight fever makes dissolving, through D10l macroporous adsorptive resins (internal diameter 1.5cm, long 15cm), water 70ml eluting; Abandon eluent, reuse 20% ethanol 70ml eluting discards eluent, continues with 60% methanol 100ml eluting; Collect eluent, evaporate to dryness, residue add methanol 2ml makes dissolving, as control medicinal material solution.Other gets ginsenoside Rg1's reference substance, adds methanol and processes the solution that every 1ml contains 1mg, as reference substance solution.According to thin layer chromatography (appendix VI B) test, draw need testing solution 10 μ l and above-mentioned control medicinal material solution and each 5 μ l of reference substance solution under the item of [discriminating] (6), put respectively on same silica gel g thin-layer plate; With 10 ℃ of lower floor's solution with held of chloroform-methanol-water (13: 7: 2) is developing solvent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.In the test sample chromatograph, with control medicinal material and the corresponding position of reference substance chromatograph on, show the speckle of same color.Collection of illustrative plates is seen Fig. 9.
Claims (18)
1. a brain-strengthening Chinese medicine composition is characterized in that, this Chinese medicine composition is processed by following parts by weight of Chinese traditional medicine:
Radix Angelicae Sinensis: 20-35; Concretio Silicea Bambusae: 10-15;
Herba Cistanches: 20-27; Dens Draconis: 10-15;
Rhizoma Dioscoreae: 20-27; Succinum: 10-15;
Fructus Schisandrae Chinensis: 15-21; Rhizoma Gastrodiae: 5-8;
Semen Platycladi: 4-6; Radix Salviae Miltiorrhizae: 5-8;
Fructus Alpiniae Oxyphyllae: 15-21; Radix Ginseng: 5-8;
Radix Polygalae: 10-15; Flos Chrysanthemi: 5-8;
Rhizoma Anemones Altaicae: 10-15; Haematitum: 7-11;
Arisaema Cum Bile: 10-15; Semen Ziziphi Spinosae: 40-55;
Fructus Lycii: 20-27;
Said Herba Cistanches is the salt Herba Cistanches (processed); Said Dens Draconis is a calcined DENS DRACONIS; Said Fructus Schisandrae Chinensis is the wine Fructus Schisandrae Chinensis (steamed); Said Semen Platycladi is for frying Semen Platycladi; Said Fructus Alpiniae Oxyphyllae is the salt stir-baked FRUCTUS ALPINIAE OXY-PHYLLAE; Said Radix Polygalae is the liquorice beverage Cortex et Radix Polygalae (processed); Said Semen Ziziphi Spinosae is a Semen Ziziphi Spinosae (parched).
2. Chinese medicine composition according to claim 1 is characterized in that, this Chinese medicine composition is processed by following parts by weight of Chinese traditional medicine:
Radix Angelicae Sinensis: 33.3; Concretio Silicea Bambusae: 13.3;
Salt Herba Cistanches (processed): 26.7; Calcined DENS DRACONIS: 13.3;
Rhizoma Dioscoreae: 26.7; Succinum: 13.3;
Wine Fructus Schisandrae Chinensis (steamed): 20; Rhizoma Gastrodiae: 6.7;
Fry Semen Platycladi: 5.3; Radix Salviae Miltiorrhizae: 6.7;
Salt stir-baked FRUCTUS ALPINIAE OXY-PHYLLAE: 20; Radix Ginseng: 6.7;
Liquorice beverage Cortex et Radix Polygalae (processed): 13.3; Flos Chrysanthemi: 6.7;
Rhizoma Anemones Altaicae: 13.3; Haematitum: 10;
Arisaema Cum Bile: 13.3; Semen Ziziphi Spinosae (parched): 53.3;
Fructus Lycii: 26.7.
3. Chinese medicine composition according to claim 1 is characterized in that, this Chinese medicine composition is processed by following parts by weight of Chinese traditional medicine:
Radix Angelicae Sinensis: 25; Concretio Silicea Bambusae: 10;
Salt Herba Cistanches (processed): 20; Calcined DENS DRACONIS: 10;
Rhizoma Dioscoreae: 20; Succinum: 10;
Wine Fructus Schisandrae Chinensis (steamed): 15; Rhizoma Gastrodiae: 5;
Fry Semen Platycladi: 4; Radix Salviae Miltiorrhizae: 5;
Salt stir-baked FRUCTUS ALPINIAE OXY-PHYLLAE: 15; Radix Ginseng: 5;
Liquorice beverage Cortex et Radix Polygalae (processed): 10; Flos Chrysanthemi: 5;
Rhizoma Anemones Altaicae: 10; Haematitum: 7.5;
Arisaema Cum Bile: 10; Semen Ziziphi Spinosae (parched): 40;
Fructus Lycii: 20.
4. according to each described Chinese medicine composition of claim 1-3, it is characterized in that said Fructus Schisandrae Chinensis is the dry mature fruit of magnoliaceae schisandra, practise and claim " Radix Schisandrae Bicoloris "; Said Rhizoma Anemones Altaicae adopts " Shandong Province's Chinese medicine processing standard 2002 " to concoct.
5. according to each described Chinese medicine composition of claim 1-3, it is characterized in that this Chinese medicine composition is pharmaceutically acceptable dosage form: tablet, pill, powder, capsule, granule and liquid agent.
6. Chinese medicine composition according to claim 5 is characterized in that, this Chinese medicine composition is capsule, pill or tablet.
7. Chinese medicine composition according to claim 6 is characterized in that, its dosage form is a capsule.
8. Chinese medicine composition according to claim 6 is characterized in that, its dosage form is a pill.
9. one kind according to each described Chinese medicine composition of claim 1-4, it is characterized in that, in every 1g Chinese medicine composition, contains Fructus Schisandrae Chinensis in schisandrin, is no less than 0.15mg.
10. Chinese medicine composition according to claim 9 is characterized in that, in every 1g Chinese medicine composition, contains Fructus Schisandrae Chinensis in schisandrin, is no less than 0.20mg.
11. the method for preparing of each described Chinese medicine composition of claim 1-4 is characterized in that, gets the raw material of Chinese medicine medicine; Haematitum, succinum, Concretio Silicea Bambusae are ground into fine powder respectively; All the other ten Six-elements are ground into fine powder, with above-mentioned fine powder facing-up, sieve; Mixing is processed capsule, powder, granule or tablet.
12. the method for preparing of each described Chinese medicine composition of claim 1-4 is characterized in that, with Haematitum, succinum, the single porphyrize powder of Concretio Silicea Bambusae difference; All the other ten Six-elements are ground into fine powder; Sieve with succinum fine powder, Concretio Silicea Bambusae fine powder, 30%-60% Haematitum fine powder, mixing adds the sodium alginate mixing with water pill or machine pill again; Polishing, drying; With Resina persicae, residue Haematitum fine powder coating, drying is processed pill.
13. method for preparing according to claim 12 is characterized in that, earlier with succinum fine powder, Concretio Silicea Bambusae fine powder, 50% Haematitum fine powder and other ten Six-elements mixing pills, then 50% Haematitum fine powder is used for coating.
14. method for preparing according to claim 11 is characterized in that, gets the raw material of Chinese medicine medicine of following weight:
Radix Angelicae Sinensis: 33.3g; Concretio Silicea Bambusae: 13.3g;
Salt Herba Cistanches (processed): 26.7g; Calcined DENS DRACONIS: 13.3g;
Rhizoma Dioscoreae: 26.7g; Succinum: 13.3g;
Wine Fructus Schisandrae Chinensis (steamed): 20g; Rhizoma Gastrodiae: 6.7g;
Fry Semen Platycladi: 5.3g; Radix Salviae Miltiorrhizae: 6.7g;
Salt stir-baked FRUCTUS ALPINIAE OXY-PHYLLAE: 20g; Radix Ginseng: 6.7g;
Liquorice beverage Cortex et Radix Polygalae (processed): 13.3g; Flos Chrysanthemi: 6.7g;
Rhizoma Anemones Altaicae: 13.3g; Haematitum: 10g;
Arisaema Cum Bile: 13.3g; Semen Ziziphi Spinosae (parched): 53.3g;
Fructus Lycii: 26.7g;
Haematitum, succinum, Concretio Silicea Bambusae are ground into fine powder respectively, and all the other ten Six-elements are ground into fine powder, with above-mentioned fine powder facing-up, sieve, and mixing, the system granule, drying, encapsulated, process 1000 capsules.
15. method for preparing according to claim 13 is characterized in that, gets the raw material of Chinese medicine medicine of following weight:
Radix Angelicae Sinensis: 25g; Zhu is yellow: 10g;
Salt Herba Cistanches (processed): 20g; Calcined DENS DRACONIS: 10g;
Rhizoma Dioscoreae: 20g; Succinum: 10g;
Wine Fructus Schisandrae Chinensis (steamed): 15g; Rhizoma Gastrodiae: 5g;
Fry Semen Platycladi: 4g; Radix Salviae Miltiorrhizae: 5g;
Salt stir-baked FRUCTUS ALPINIAE OXY-PHYLLAE: 15g; Radix Ginseng: 5g;
Liquorice beverage Cortex et Radix Polygalae (processed): 10g; Flos Chrysanthemi: 5g;
Rhizoma Anemones Altaicae: 10g; Haematitum: 7.5g;
Arisaema Cum Bile: 10g; Semen Ziziphi Spinosae (parched): 40g;
Fructus Lycii: 20g;
With Haematitum, succinum, the single porphyrize powder of Concretio Silicea Bambusae difference, all the other ten Six-elements are ground into fine powder, sieve with succinum fine powder, Concretio Silicea Bambusae fine powder, 50% Haematitum fine powder again, and mixing adds the sodium alginate mixing with water pill or machine pill, polishing, drying; With Resina persicae, residue Haematitum fine powder coating, drying is processed pill.
16. the quality determining method of each described Chinese medicine composition of claim 1-4 is characterized in that, this quality determining method comprises microscopical identification and thin layer chromatography discrimination method:
(1) microscopical identification: get Chinese medicine composition, put microscopically and observe: plant skin palisade cells brownish red, norma lateralis is the string palisade to be arranged, and polygon is seen on the surface, and cell is big slightly, includes the dark-brown thing; The pale brown color of endotesta cell, surface see rectangle or class is square, and the anticlinal wall beaded thickens; Plant the pale brown color of skin epidermis stone cell, a type polygon is seen on the surface, and wall is thicker, and the hole ditch is fine and closely woven, and cell contains the burgundy thing; Plant skin stone cell surface and see irregular polygon, wall thickness, wavy bending, laminated striation is clear;
(2) Radix Angelicae Sinensis characteristic thin layer chromatography is differentiated: get Chinese medicine composition 4g, porphyrize adds ethanol 40ml, and supersound process 20 minutes filters, and filtrating is concentrated into about 2ml, as need testing solution; Other gets Radix Angelicae Sinensis control medicinal material 1g, adds ethanol 10ml, shines medical material solution in pairs with legal system; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be that 9: 1 developing solvent launches with cyclohexane extraction-ethyl acetate, take out, dry, put under the ultra-violet lamp 365nm and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color;
(3) get Chinese medicine composition 5g, porphyrize adds water 100ml, and heated and boiled 15 minutes is put coldly, centrifugal, gets supernatant, adds ethyl acetate extraction 3 times, each 50ml, and combined ethyl acetate liquid, evaporate to dryness, residue add ethyl acetate 2ml makes dissolving, as need testing solution; Other gets Fructus Lycii control medicinal material 1g, shines medical material solution in pairs with legal system; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be that 6: 4: 0.2 developing solvent launches with toluene-ethyl acetate-formic acid, take out, dry, put under the ultra-violet lamp 365nm and inspect;
(4) get Chinese medicine composition 10g, porphyrize, the 60ml that adds diethyl ether, supersound process 30 minutes is put coldly, filters; Discard ether solution, residue volatilizes solvent, adds methanol 100ml, and supersound process 30 minutes is put cold; Filter, the filtrating evaporate to dryness, residue adds water 10ml slight fever makes dissolving, through the D101 macroporous adsorptive resins, its internal diameter 1.5cm; Long 15cm, water 70ml eluting discards eluent, and reuse 20% ethanol 70ml eluting discards eluent; Continue with 60% methanol 100ml eluting, collect eluent, evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the echinacoside reference substance, adds methanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to thin layer chromatography test, draw each 2 μ l of above-mentioned two kinds of solution, put respectively on same polyamide film, be that 2: 1: 7 developing solvent launches with methanol-acetic acid-water, take out, dry, put under the ultra-violet lamp 365nm and inspect;
(5) get Fructus Schisandrae Chinensis control medicinal material 1g, add ethanol 10ml, supersound process 20 minutes filters, and filtrating is concentrated into about 2ml, as control medicinal material solution; Get schisandrin B reference substance, schisandrin reference substance again, add methanol respectively and process the solution that every 1ml contains 1mg, as reference substance solution; Test according to thin layer chromatography; Draw need testing solution 10 μ l and above-mentioned control medicinal material solution and each 5 μ l of reference substance solution under discrimination method (2) item; Putting respectively on same silica GF254 lamellae, is that 7: 2.5 developing solvent launches with cyclohexane extraction-ethyl acetate, takes out; Dry, put under the ultra-violet lamp 254nm and inspect;
(6) get need testing solution under the discrimination method (4), evaporate to dryness, residue add water 15ml makes dissolving; Extract 3 times with water saturated n-butyl alcohol jolting, each 15ml merges n-butyl alcohol liquid; With ammonia solution washing 2 times, each 15ml, n-butyl alcohol liquid evaporate to dryness; Residue adds methanol 1ml makes dissolving, as need testing solution; Other gets jujuboside A and jujuboside B reference substance, adds methanol and processes the mixed solution that every 1ml contains 1mg, as reference substance solution; According to the thin layer chromatography test, draw above-mentioned need testing solution 10 μ l and reference substance solution 5 μ l, put respectively on same silica gel g thin-layer plate, be developing solvent with water saturated n-butyl alcohol, launch, take out, to dry, spray is inspected with 2% vanillin sulfuric acid solution immediately; In the test sample chromatograph, with the corresponding position of reference substance chromatograph on, show the speckle of same color;
(7) get Radix Ginseng control medicinal material 1g, add methanol 50ml, supersound process 30 minutes is put coldly, filters; The filtrating evaporate to dryness, residue adds water 10ml slight fever makes dissolving, through the D101 macroporous adsorptive resins, its internal diameter 1.5cm, long 15cm; Water 70ml eluting is abandoned eluent, and reuse 20% ethanol 70ml eluting discards eluent, continues with 60% methanol 100ml eluting; Collect eluent, evaporate to dryness, residue add methanol 2ml makes dissolving, as control medicinal material solution; Other gets ginsenoside Rg1's reference substance, adds methanol and processes the solution that every 1ml contains 1mg, as reference substance solution; According to the thin layer chromatography test, draw need testing solution 10 μ l and above-mentioned control medicinal material solution and each 5 μ l of reference substance solution under discrimination method (6) item, put respectively on same silica gel g thin-layer plate; With chloroform-methanol-water be 13: 7: 2,10 ℃ of lower floor's solution with held are developing solvent; Launch, take out, dry; Spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃.
17. the effect of each described Chinese medicine composition of claim 1-8 in the medicine of preparation treatment nourishing the brain and improving intelligence, hypnotic, resisting fatigue, cold-resistant, anoxia enduring, enhancing humoral immunization.
18. application according to claim 17 is characterized in that, said application is at preparation treatment brain overwork, amnesia, and the neurasthenia has a dizzy spell, and the palpitation with fear insomnia vexedly is prone to tiredly, and the fear of cold body is empty, and body loses the application in the medicine of pain in the lumbar region and senile dementia.
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| CN102846755B (en) * | 2012-03-23 | 2014-04-09 | 曲延丽 | Traditional Chinese medicine for treating cerebral concussion sequelae |
| CN103048410B (en) * | 2013-01-24 | 2014-09-24 | 贵州百灵企业集团制药股份有限公司 | Detection method of thin membrane coated tablet for treating sleep disorders |
| CN103300148A (en) * | 2013-05-16 | 2013-09-18 | 吴永金 | Fruity brain strengthening granule and preparation method thereof |
| CN104288396A (en) * | 2014-09-27 | 2015-01-21 | 隋荣玲 | Traditional Chinese medicine decoction for treating senile dementia |
| CN104833661A (en) * | 2015-01-30 | 2015-08-12 | 吉林修正药业新药开发有限公司 | Quality control method of drug for nourishing blood and quieting spirit |
| CN105250642A (en) * | 2015-10-28 | 2016-01-20 | 雷少华 | Traditional Chinese medicine composition for treating insomnia |
| CN108210818B (en) * | 2018-04-16 | 2021-03-02 | 成都中医药大学 | Pharmaceutical composition for preventing and treating neurodegenerative diseases and preparation method and application thereof |
| CN110478456B (en) * | 2019-09-18 | 2022-02-22 | 上海医药集团青岛国风药业股份有限公司 | Preparation method of brain-strengthening medicinal pills |
| CN112083090B (en) * | 2020-08-27 | 2022-03-22 | 四川新绿色药业科技发展有限公司 | Thin-layer identification method for traditional Chinese medicine preparation containing concretio silicea bambusae or artificial concretio silicea bambusae |
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