CN102085282B - Traditional Chinese medicine composition for treating hypophrenia and senile dementia and preparation method and detection method thereof - Google Patents
Traditional Chinese medicine composition for treating hypophrenia and senile dementia and preparation method and detection method thereof Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine composition for treating hypophrenia and senile dementia and a preparation method and detection method thereof. The traditional Chinese medicine composition is prepared from ginseng, rhizoma acori graminei, chuanxiong rhizome, ginkgo leaf, prepared rehmannia root and senega, can be prepared into pills, granules, hard capsules, soft capsules, tablets, dripping pills, oral liquid or the like, has the characteristics of exact treatment effect, few side effects and low price, and can better improve clinical symptoms. The detection method of the oral liquid comprises the following steps: carrying out thin layer chromatography on rhizoma acori graminei, chuanxiong rhizome, ginseng and senega; inspecting the relative density and pH value; and measuring the content of ferulic acid.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition and method for making thereof and detection method, particularly a kind of Chinese medicine composition and method for making and detection method of treating mental retardation and senile dementia.
Background technology
Dementia is the acquired and persistent syndrome of intelligent disorder that produces because of disordered brain function.Senile dementia is a kind of chronic progressive external mental deterioration property disease, and along with the raising of living standards of the people, the significantly increase in life-span, and the quickening of aged tendency of population trend and significantly increasing keeps healthy with seniculture to socio-economic development and has brought huge challenge." the Chinese experimental pharmacology of Chinese medical formulae " magazine the 16th the 9th phase of volume of August in 2010 " brain-strengthening is given birth to the effect of intelligence oral liquid to the senile dementia mouse model " works such as () Pang Laixiang discloses a kind of Chinese medicine of treating mental retardation and senile dementia---brain-strengthening life intelligence oral liquid; Process by Radix Ginseng, Rhizoma Acori Graminei, Rhizoma Chuanxiong, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae; But concrete proportioning raw materials is not disclosed; On the basis of original achievement, we have found the proportioning of science through practice, and are evident in efficacy.
Summary of the invention
The object of the present invention is to provide a kind of existing definite curative effect, can improve the Chinese medicine composition of clinical symptoms, few side effects, low-cost treatment mental retardation and senile dementia again preferably.
Another purpose of the present invention is to provide the method for preparing of this Chinese medicinal composition preparation.
Another object of the present invention is to provide the detection method of this Chinese medicine composition oral liquid.
Technical scheme of the present invention is achieved in that
According to the weight portion meter, the raw material of processing this Chinese medicine composition active component is: Radix Ginseng 190-310, Rhizoma Acori Graminei 190-310, Rhizoma Chuanxiong 190-310, Folium Ginkgo 190-310, Radix Rehmanniae Preparata 225-375, Radix Polygalae 110-190.
Be preferably: Radix Ginseng 220-280, Rhizoma Acori Graminei 220-280, Rhizoma Chuanxiong 220-280, Folium Ginkgo 220-280, Radix Rehmanniae Preparata 260-340, Radix Polygalae 130-170.
Most preferably be: Radix Ginseng 250, Rhizoma Acori Graminei 250, Rhizoma Chuanxiong 250, Folium Ginkgo 250, Radix Rehmanniae Preparata 300, Radix Polygalae 150.
Chinese medicine composition of the present invention has benefiting QI for activating blood circulation, kidney tonifying, essence replenishing, the effect of eliminating the phlegm and having one's ideas straightened out.Be used to suffer from a deficiency of the kidney that marrow subtracts, the mental retardation due to the expectorant stasis of blood resistance key and the treatment of senile dementia.
Chinese medicine composition of the present invention can be an acceptable forms on the pharmaceutics, comprises pill, granule, hard capsule, soft capsule, tablet, drop pill, oral liquid etc., preferred oral liquid.
Rhizoma Chuanxiong, Rhizoma Acori Graminei two flavor distillating extracting oils; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae are carried out decocting; Add proper auxiliary materials, process preparation.
Adjuvant among the present invention can be acceptable any excipient or a carrier on the pharmaceutics.
Method for preparing is preferred: Rhizoma Chuanxiong, Rhizoma Acori Graminei add 8-12 times of water gaging distillation 2-9 hour, and extraction volatile oil is subsequent use or enclose is dry, and debris device is in addition collected; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 8-12 times of water gaging and decoct 1-3 time, and each 1-2 hour, merging filtrate and above-mentioned debris; Filter, being evaporated to 20 ℃ of relative densities is 1.00-1.20, adds ethanol and makes and contain the alcohol amount and reach 40-70%; Left standstill 8-30 hour, and got supernatant, reclaiming ethanol and being concentrated into 20 ℃ of relative densities is 1.00-1.10; Left standstill 12-30 hour, and got supernatant filtration or concentrate drying, adding volatile oil and proper auxiliary materials, process preparation.
Wherein oral liquid is preferred again: Rhizoma Chuanxiong, Rhizoma Acori Graminei add 10 times of water gaging distillations 6 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 10 times of water gagings and decoct three times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered; Being evaporated to 20 ℃ of relative densities is 1.08-1.10, adds ethanol and makes and contain alcohol amount and reach 50%, leaves standstill 24 hours, gets supernatant; Reclaiming ethanol and being concentrated into 20 ℃ of relative densities is 1.06, leaves standstill 24 hours, gets supernatant and filters, and filtrating adds above-mentioned volatile oil, an amount of solubilizing agent, correctives, antiseptic etc.; Filter, packing, sterilization promptly gets.
Particularly: Radix Ginseng 250g, Rhizoma Acori Graminei 250g, Rhizoma Chuanxiong 250g, Folium Ginkgo 250g, Radix Rehmanniae Preparata 300g, Radix Polygalae 150g; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 10 times of water gaging distillations 6 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 10 times of water gagings and decoct three times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered, and being evaporated to 20 ℃ of relative densities is 1.08-1.10; Add ethanol and make and contain alcohol amount and reach 50%, left standstill 24 hours, get supernatant, reclaiming ethanol and being concentrated into 20 ℃ of relative densities is 1.06; Left standstill 24 hours, and got supernatant and filter, filtrating adds above-mentioned volatile oil, tween 80 and an amount of correctives etc., and the adjustment volume is to 1000ml; Filter, packing, sterilization promptly gets.
The detection method of the oral liquid that makes for last method comprises: the thin layer of Rhizoma Acori Graminei, Rhizoma Chuanxiong, Radix Ginseng, Radix Polygalae differentiates that the inspection of relative density, pH value, content of ferulic acid are measured.
One, character: the present invention is the sepia supernatant liquid; The sweet little hardship of distinguishing the flavor of.
Two, differentiate:
(1) thin layer of Rhizoma Acori Graminei is differentiated: get 8ml of the present invention, with extracted with diethyl ether 2 times, 20ml obtains ether layer at every turn, and evaporate to dryness, residue add ethyl acetate 1ml, make dissolving, as need testing solution.Other gets Rhizoma Acori Graminei control medicinal material 2g, the 25ml that adds diethyl ether, and supersound process 10min filters, the filtrating evaporate to dryness, residue adds ethyl acetate 1ml, makes dissolving, as control medicinal material solution.According to thin layer chromatography (2010 version " an appendix VI of Chinese pharmacopoeia B) test, draw each 5 μ l of above-mentioned 2 kinds of solution, putting in the same 0.5%CMC-Na of containing solution respectively is on the silica gel g thin-layer plate of binding agent; With petroleum ether (60-90 ℃)-ethyl acetate (8: 2) is developing solvent, launches, and exhibition is apart from 8cm; 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 identical fluorescence speckle; Smoked clear with iodine vapor again to the speckle colour developing, inspect under the daylight, show identical sepia speckle.
(2) thin layer of Rhizoma Chuanxiong is differentiated: get 8ml of the present invention, with extracted with diethyl ether 2 times, 20ml obtains ether layer at every turn, and evaporate to dryness, residue add ethyl acetate 1ml, make dissolving, as need testing solution.Other gets Rhizoma Chuanxiong control medicinal material 2g, the 25ml that adds diethyl ether, and supersound process 10min filters, and gets the filtrating evaporate to dryness, and residue adds ethyl acetate 1ml makes dissolving, as control medicinal material solution.According to thin layer chromatography (2010 version " an appendix VI of Chinese pharmacopoeia B) test, draw each 10 μ l of above-mentioned 2 kinds of solution and put respectively on same silica gel g thin-layer plate, be developing solvent with normal hexane-ethyl acetate (9: 1); Launch, exhibition is taken out apart from 8cm; 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 identical fluorescence speckle.
(3) thin layer of Radix Ginseng is differentiated: get 2mL of the present invention, thin up is to 15ml, with water saturated n-butanol extraction 2 times, and 25mL at every turn; Merge butanol extraction liquid, n-butyl alcohol liquid is with 0.1%NaOH solution washing 2 times, 15mL at every turn; The saturated water washing of reuse n-butyl alcohol 2 times, each 20mL abandons or adopts water layer; N-butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methanol 1mL dissolving, as need testing solution.Other get the reference substance ginsenoside Rb1 add methanol process respectively contain 1mgmL-1 mixed solution as reference substance solution.According to thin layer chromatography (2010 version " an appendix VI of Chinese pharmacopoeia B) test, draw each 10 μ L of above-mentioned 2 kinds of solution, put respectively on same silica gel g thin-layer plate; (15: 40: 22: 10) 10 ℃ of lower floor's solution with held were developing solvent with chloroform ethyl acetate-methanol-water; Launch, take out airing; Spray is dried by the fire to clear spot in 105 ℃ with 10% ethanol solution of sulfuric acid.Put respectively under daylight and the ultra-violet lamp (365nm) and inspect, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the speckle of same color.
(4) thin layer of Radix Polygalae is differentiated: get 10ml of the present invention, divide 2 extractions with ether, each 20ml merges ether solution, washes 3 times, each 30ml, and ether solution low temperature evaporate to dryness, residue add ethyl acetate 0.5ml makes dissolving, as need testing solution.Other gets Radix Polygalae control medicinal material 1.5g, adds water 50ml and decocts 1h, filters, and filtrating is divided 2 extractions with ether, and merge extractive liquid, is concentrated into 1ml, as control medicinal material solution.According to thin layer chromatography (2010 version " an appendix VI of Chinese pharmacopoeia B) test; Drawing each 4 μ l of above-mentioned 2 kinds of solution puts respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; (30: 15: 5: 0.5) be developing solvent, expansion was taken out with cyclohexane extraction-chloroform-ethyl acetate-methanol; 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.
Three, inspection:
1, relative density: should be not less than 1.05 (" an appendix VII of Chinese pharmacopoeia version in 2010 A).
2, pH value: should be 4.0-6.0 (2010 version " an appendix VII of Chinese pharmacopoeia G).
3, other: should meet pertinent regulations under the mixture item (" an appendix I of Chinese pharmacopoeia version in 2010 J).
Four, assay
Measure according to HPLC (" an appendix VI of Chinese pharmacopoeia version in 2010 D).
Chromatographic condition and system suitability test: use octadecylsilane chemically bonded silica to be filler; Methanol-1% glacial acetic acid (28: 72) is a mobile phase; The detection wavelength is 321nm; Theoretical cam curve is pressed the ferulic acid peak and is calculated, and should be not less than 2000.
The preparation of reference substance solution: it is an amount of that precision takes by weighing the ferulic acid reference substance, adds methanol and process the solution that every ml contains 12 μ g, as reference substance solution.
The preparation of need testing solution: accurate absorption 1ml of the present invention, put in the volumetric flask of 25ml, add mobile phase and be diluted to scale, shake up, as need testing solution.
Algoscopy: accurate respectively reference substance solution and each 5ul of need testing solution of drawing, inject chromatograph of liquid, measure, promptly get.
The every ml of the present invention contains Rhizoma Chuanxiong with ferulic acid (C
10H
10O
4) meter, should be not less than 250ug.
Five, usage and dosage: oral, a 20ml, 2 times on the one.
Six, specification: every dress 20ml.
Slow-witted person is crazy about also, and also insane, also not intelligent, the meaning of lacking common sense also.Back physicians from a family for generations family is according to its characteristics appellation dementia again.Mainly show as dysmnesia, hypophrenia, the behavior personality change, severe patient can be lost self care ability, belongs to " melancholia ", " dementia ", " demented " category more.Primary disease is a kind of systemic disease, and sick position is at brain, and is closely related with the imbalance of conscience spleen renal function, the cause of disease with interior because main.Since internal injury caused by excess of seven emotions, the prolonged illness consume, body deficiency due to old age causes gas, blood, expectorant, the stasis of blood, strongly fragrant for suffering from, and gradually makes brains hollow, or insufficiency of vital energy and blood, the kidney essense loss, and the mutual resistance of the expectorant stasis of blood, it is primary disease that brains lose hair care." kidney be the vital organs of the human body this, the root of vigour ", kidney storing essence is given birth to marrow, deficiency of kidney-essence can not be given birth to marrow, " the soreness of the shank dizziness, order does not have finding, slack peacefully sleeping for deficiency of marrow-reservoir, vertigo and tinnitus then ".With age, be that master's the five internal organs void declines and takes place gradually to suffer from a deficiency of the kidney, cause mechanism of qi unsmooth unfavorable, the body fluid interruption of service, and give birth to the expectorant stasis of blood." dementia is all does not have expectorant usually, and or with pent-up, or with unsuccessful ... gradually cause dementia ", " control slow-witted strange method, control expectorant promptly control stay also ".Mostly senile dementia patient body of the tongue is that purple is dark, dim or petechia, ecchymosis are arranged, and the occurrence rate of greasy fur also is the highest, has also confirmed the existence of blood stasis and phlegm-damp.Clinically control and receive better curative effect from the blood stasis opinion, explain that also blood stasis is influence a brain aging and a key factor that causes the senile dementia generation diseases such as cerebral arteriosclerosis, cerebrovascular dementia are many.In a word, senile dementia is to be person in middle and old age's frequently-occurring disease of characteristic with the deficiency in origin and excess in superficiality, and its deficiency in origin mainly is deficiency of kidney-essence, and brain is lost empty, clear YANG failing to ascend; Its mark is turbid in expectorant really, blood stasis is hoodwinked the brain key, impatency brain network.
The present invention treats senile dementia and forms based on the heightened awareness prescription to its etiology and pathogenesis.Radix Ginseng strongly invigorating primordial QI in the side makes the prosperous then cloudy blood of gas spontaneous, and the Fructus Alpiniae Oxyphyllae of calming the nerves again, is monarch drug.Shennong's Herbal: " tonifying five ZANG-organs, peace spirit is decided soul, spasmolytic is throbbed with fear, and removes pathogen, makes eye bright, happy Fructus Alpiniae Oxyphyllae ".The Rhizoma Chuanxiong acrid in the mouth is warm in nature, can go the Shu Yu that looses, kind again promoting blood circulation, and fragrance is walked to scurry, and can rise to fall; Be good at the up head, go into brain and open and close, also be the key medicine of current QI and blood, " book on Chinese herbal medicine converge speech " pointed out: " the up head of Rhizoma Chuanxiong; following regulating menstruation water, middle relieving stagnation, gas medicine in the blood ... acrid in the mouth property sun; scurry under gas is kind and depletion of YIN coagulates viscid attitude, though go into blood system, can go a wind cutting again, transfer all gas "; The Radix Rehmanniae Preparata quality is soft and moist, and sweet in the mouth is and thick, warm in nature not dry, and it is Kidney-Yin to be longer than nourishing, promoting generation of vital essence and blood; Fill out bone marrow, fill brain, hearing-improing and eyesight improving, the Fructus Alpiniae Oxyphyllae of reposing " medicineization justice " to being summarized as of its effect " nourish Kidney-Yin, packing bone marrow; for panacea also ", two medicinal be minister, with the monarch drug compatibility, can make blood capable fluent, the brain ruton is smooth; Smart superior and fill marrow, blood gets up and foster brain,, play blood circulation promoting and blood stasis dispelling, the effect of the refreshment of having one's ideas straightened out.The hot temperature of Rhizoma Acori Graminei, fragrant sharp key, the expectorant of kind a surname's gas, the refreshment of having one's ideas straightened out, Shennong's Herbal is claimed its " happy hole, logical nine orifices ", the Sun Simiao speech: " clothes are made light of one's life by commiting suicide for a long time, and hearing-improing and eyesight improving is not forgotten, and does not confuse, beneficial intelligence, high will is not old "; Radix Polygalae is arduous to be slightly cold, and being longer than eliminates the phlegm has one's ideas straightened out, the beneficial will of calming the nerves, and it be " Fructus Alpiniae Oxyphyllae is intelligent, normal hearing and eyesight; do not forget " that Shennong's Herbal is summed up its effect, " property of medicine opinion " pointed out: " control forgetfully, pacify soul, make us not being confused "; Rhizoma Acori Graminei is hot partially to loose to declare its phlegm-damp, and the expectorant that the inclined to one side hardship of Radix Polygalae is fallen to let out superinverse stops up, and Folium Ginkgo property sweetness and bitterness is puckery, has that the beneficial heart is astringed the lung, an effect of removing dampness antidiarrheal, blood circulation promoting and blood stasis dispelling, TONGMAI SHULUO; Three medicines for assistant, are usefulness to help the monarch and his subjects altogether, make gas along and stop up from opening, QI and blood is not gone up aster again; Clear key is not covered in the turbid dissipation of expectorant, and mind is from can be clear and bright, and the up head of Rhizoma Chuanxiong is that ministerial drug and the messenger drug of holding concurrently are for also.The all medicine compatibilities in full side, the merit of playing benefiting QI for activating blood circulation, kidney tonifying, essence replenishing altogether, eliminating the phlegm and having one's ideas straightened out, mental retardation and the senile dementia that marrow subtracts, the expectorant stasis of blood hinders due to the key that be used to suffer from a deficiency of the kidney has good effect.If can informal dress, can protect clear-headed and clear-sighted, refreshing know salubrious, wisdom is intelligent, send out Yan Rutong, does not so then have the anxiety of forgetful dementia.
Chinese medicine preparation main pharmacodynamics experimental result of the present invention is following: with oral liquid of the present invention is example.
One, receive the reagent thing:
Radix Ginseng 250g, Rhizoma Acori Graminei 250g, Rhizoma Chuanxiong 250g, Folium Ginkgo 250g, Radix Rehmanniae Preparata 300g, Radix Polygalae 150g; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 10 times of water gaging distillations 6 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 10 times of water gagings and decoct three times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered; 20 ℃ are evaporated to relative density 1.08-1.10, add ethanol and make and contain alcohol amount and reach 50%, leave standstill 24 hours, get supernatant; Reclaim ethanol and 20 ℃ and be concentrated into relative density 1.06, left standstill 24 hours, get supernatant and filter, filtrating adds above-mentioned volatile oil; Filter, sterilization promptly gets.
Two, the test of pesticide effectiveness
1, scopolamine is caused the influence of Senile Dementia Model Rats brain AchE, brain protein content
1.1 method is got 60 of rats, male and female half and half are divided into 6 groups at random by body weight, are respectively the normal control group, model group, positive piracetam group (0.3gkg
-1), high dose group (16.8gkg of the present invention
-1), middle dose groups (8.4gkg
-1), low dose group (4.2gkg
-1), normal group, model group give the equal-volume distilled water.Irritate the long-pending 10mLkg of being of body of stomach
-1, 1 day 1 time, continuous 10d.Begin lost training simultaneously, 20min before training, except that normal group, each organizes lumbar injection scopolamine 2mgkg
-1, train 9d continuously.2h after the last administration, dislocation method put to death animal, take out rat cerebral tissue's (removal XIAONAO) fast, put rinsing in the pre-cooling normal saline, and filter paper is wiped away dried, after weighing, immerse in the cold saline (for 9 times of tissue weight) rapidly, in ice bath, process 100mgmL with homogenizer
-1Tissue homogenate, with 3000rmin
-1, centrifugal 10min gets supernatant and is used to measure AchE, brain protein content.
Obviously raise 1.2 the results model group AchE of rat cerebral tissue is active,, significant difference (P＜0.05) is arranged with normal group; The present invention is high, the AchE activity of middle dose groups also significantly reduces, and with model group, significant difference (P＜0.05) is arranged; Low dose group has the active trend of the AchE of reduction; The therapeutical effect of prompting piracetam and anti-senile dementia of the present invention is to realize through the effect that reduces AchE activity, rising Ach content and raising central cholinergic system.
Model group rat brain protein content obviously reduces, and with normal group significant difference (P＜0.05) is arranged; Compare the equal showed increased of brain protein content (P＜0.05) of height of the present invention, middle dose groups rat with model group; Show that the present invention maybe be synthetic relevant with brain albumen to the influence of the learning and memory function of senile dementia.
Table 1 pair scopolamine causes the influence (
n=10) of Senile Dementia Model Rats brain AchE, brain protein content
Annotate: compare P＜0.05 with model group
2, to AlCl
3Cause the influence of the learning and memory function of Senile Dementia Model Rats
2.1 method is got 60 of rats, male and female half and half are removed positive brain-invigorating capsule group (0.162gkg
-1) outside, divide into groups with 2.1 projects.Fasting 8h before the modeling, except that the normal control group, lumbar injection AlCl next day that each being organized
3100mgkg
-1, continuous 50d.20d after the modeling, gastric infusion are 10mLkg
-1, 1 day 1 time, be total to 30d.20min after the last administration measures the rat memory function through Mirros water maze method.After memory function is measured end back administration next day 2h, put to death 4 rats with each group of dislocation method, carry out pathological observation.Take out complete brain and use 100mgmL
-1Formalin is fixed, the routine paraffin wax section, and per 5 of crown serial section hippocampal tissue is got 1.Adopt HE dyeing and reticular fiber staining respectively, optical microscope is observed cerebral hippocampal structure senile plaque down, NFT changes.Remaining rat peels off cerebral tissue fast, claims its weight in wet base, calculates the heavy index of cutaneous horn (the heavy index of cutaneous horn=cutaneous horn weight/body weight * 100).Brain (removal XIAONAO) is processed 100mgmL under 2.1
-1Tissue homogenate is with 3000rmin
-1, centrifugal 10min gets supernatant and measures β-AP content.The result is carried out the t check.
2.2 each group compares zero difference before the modeling as a result; Model group and blank control group after the modeling have significant difference (P＜0.05).Respectively organize the used time of water maze after the medication and shorten than model group, wherein brain-invigorating capsule group, the high, medium and low dose groups of the present invention can obviously shorten the used time, with model group, significant difference (P＜0.05) are arranged.Explain: the present invention has the senile dementia of enhancing learning and memory function.See table 2 for details
Model group and blank control group compare, and the heavy index of cutaneous horn obviously reduces, and significant difference (P＜0.05) is arranged.The present invention is high, middle dose groups obviously raises, and heavy index of cutaneous horn and model group have significant difference (P＜0.05); The heavy index of low dose group cutaneous horn of the present invention all increases.Explain: the present invention is better to improving senile dementia rat brain weight in wet base index effect.
Model group and blank control group compare, and β in the cerebral tissue-AP content obviously increases, and significant difference (P＜0.05) is arranged.Each treatment group is compared with model group, and β in the cerebral tissue-AP content all has decline in various degree, and wherein the high, medium and low dose groups of the present invention can obviously reduce the β-AP content in the cerebral tissue, has compared significant difference (P＜0.05) with model group.Explain: thus the present invention maybe β-AP content reaches the effect of anti-senile dementia in the senile dementia rat cerebral tissue through reducing.See table 3 for details.
Table 2 couple AlCl
3Cause the effect of Senile Dementia Model Rats water maze influence (
N=10)
Annotate: compare with model group,
#P＜0.05
Table 3 couple AlCl
3Cause the heavy index of Senile Dementia Model Rats cutaneous horn, β-AP content influence (
N=10)
Annotate: compare with model group,
#P＜005
The pathological section result:
Blank control group: pyramidal cell shows clearly in the hippocampus brain tissue slice, and form is normal, does not see NFT and senile plaque.Model group: can know under the hippocampus brain tissue slice HE dyeing high power lens and see a large amount of NFTs; Can know under the reticular fiber staining high power lens and see several senile plaques, the modeling success is described.
The brain-invigorating capsule group: the down visible more preceding obvious minimizing of NFT number of hippocampus brain tissue slice HE dyeing high power lens, in the visual field, do not see senile plaque, explain that brain-invigorating capsule can reduce the number of the person in middle and old age of senile dementia rat cerebral tissue speckle.
The present invention is high, middle dose groups: hippocampus brain tissue slice HE dyeing and reticular fiber staining high power lens downward view inner cone cell clear display, all do not see NFT and senile plaque; Can see several senile plaques under the hippocampus brain tissue slice reticular fiber staining high power lens of low dose group, explain that the present invention is high, middle dose groups has the effect that reduces NFT and senile plaque number in the senile dementia rat hippocampus district cerebral tissue.
3, to alzheimer disease rat model cerebral tissue SOD, MDA content, Na
+-k
+The influence of-atpase activity
3.1 method is got 60 of rats, ♀ is divided into 6 groups at random by body weight, 10 every group, is respectively the normal control group, model group, positive piracetam group (0.301gkg
-1), high dose group (16.8gkg of the present invention
-1), middle dose groups (8.4gkg
-1), low dose group (4.2gkg
-1), normal group, model group give the equal-volume distilled water, and the administration volume is 10mLkg
-1, once a day, continuous 10d.Begin lost training simultaneously, 20min before training, except that normal group, each organizes lumbar injection scopolamine 2mgkg
-1, train 9d continuously, 2h after the last administration; Dislocation method is put to death animal, takes out rat cerebral tissue's (removal XIAONAO) fast, puts rinsing in the pre-cooling normal saline; Filter paper is wiped away dried, after weighing, immerses in the cold saline (for 9 times of tissue weight) rapidly; In ice bath, process 10% tissue homogenate with homogenizer, with 3000rmin
-1, centrifugal 10min gets supernatant and is used to measure SOD activity, active, the Na of MDA
+-k
+-atpase activity.
Obviously reduce 3.2 the results model group SOD of rat cerebral tissue is active, compare, significant difference (P＜0.05) is arranged with normal group; Piracetam group and the present invention are high, middle dose groups SOD activity raises gradually and is dose dependent; Compare with model group; Significant difference (P＜0.05) is all arranged; High dose group SOD value of the present invention has been compared significant difference (P＜0.05) apparently higher than the piracetam group with the piracetam group, prompting the present invention is superior to piracetam.Show that the present invention possibly be through SOD activity improving to the therapeutical effect that improves of alzheimer disease, the brain tissue oxygen free radical is reduced and realize.
The MDA of model group rat cerebral tissue is active obviously to raise, and compares with normal group, and significant difference (P＜0.05) is arranged; The high, medium and low dose groups of piracetam group and the present invention MDA of rat cerebral tissue content obviously reduces, and compares with model group, and significant difference (P＜0.05) is arranged; The high dose group MDA of rat cerebral tissue content of the present invention obviously reduces, and has compared significant difference (P＜0.05) with the piracetam group, and prompting the present invention is excellent than the piracetam group.Show that the present invention is relevant with the MDA activity to the influence of senile dementia patient learning and memory function.
The Na of model group
+-K
+-atpase activity obviously reduces, and compares with normal group, and significant difference (P＜0.05) is arranged; The high, medium and low dose groups of piracetam and the present invention can obviously improve Na
+-K
+-atpase activity relatively has significant difference (P＜0.05) with model group.High dose group Na of the present invention
+-K
+-atpase activity compares with the piracetam group apparently higher than the piracetam group, and significant difference (P＜0.05) is arranged, and prompting the present invention is superior to piracetam.Show influence and the Na of the present invention to alzheimer disease
+-K
+-atpase activity is relevant.
Table 4 couple alzheimer disease rat model cerebral tissue SOD, active, the Na of MDA
+-K
+The influence of-atpase activity (
N=10)
Annotate: compare with model group,
#P＜0.05;
4, the influence of, brain protein content active to AchE in alzheimer disease rat model blood and the brain
4.1 method is got 60 of rats, ♀ ♂ half and half removes positive brain-invigorating capsule group (0.162gkg
-1) outside, it is the same to divide into groups.Fasting 8h before the modeling, except that the normal control group, lumbar injection 1%AlCl next day that each being organized
3100mgkg
-1, continuous 50d.20d after the modeling, gastric infusion are 10mLkg
-1, once a day, be total to 30d.2h after the last administration gets brain (removal XIAONAO) and under 2.1, processes 10% tissue homogenate, with 3000rmin
-1, centrifugal 10min gets supernatant and measures brain AchE activity, brain protein content.Adopt the spss12.0 statistical software to handle the result, carry out the t check, the comparable group differences.
Obviously raise 4.2 the results model group AchE of rat cerebral tissue is active,, significant difference (P＜0.05) is arranged with normal group; The AchE activity of the high, medium and low dose groups of the present invention also significantly reduces, and with model group, significant difference (P＜0.05) is arranged; The therapeutical effect of prompting piracetam and anti-senile dementia of the present invention is to realize through the effect that reduces AchE activity, rising Ach content and raising central cholinergic system.
Total protein content obviously reduces in model group and the blank control group comparison cerebral tissue, and significant difference (P＜0.05) is arranged; Can obviously the raise content of total protein in the cerebral tissue of the present invention's height, middle dose groups, brain-invigorating capsule group has been compared significant difference (P＜0.05) with model; Low dose group of the present invention can increase the brain protein content, but compares not statistically significant (P＞0.05) with model group.Explain the present invention to anti-senile dementia maybe be in rising rat cerebral tissue total protein content relevant.
Table 5 couple alzheimer disease rat model brain AchE is active, the influence (
n=10) of brain protein content
Annotate: compare with model group,
#P＜0.05;
5, to senile dementia model mice learning and memory function and cerebral tissue SOD, MAO, the active influence of Na+-K+-ATPase
5.1 60 of healthy adult ICR mices are got in grouping and administration, are divided into 6 groups at random, are respectively normal control group, model control group, piracetam group, the high, medium and low dose groups of the present invention.All the other respectively organize the scl%D-of nape portion galactose 140mgkg every day except that normal control group injection equivalent normal saline
-1, totally 6 weeks.The back ig administration of 2 week of modeling, piracetam group (0.6gkg
-1), the high, medium and low dose groups of the present invention (is respectively 25,12.5,6.25g crude drug kg
-1), the distilled water of normal group, model group ig equivalent, totally 4 weeks.
5.2 index determining
5.2.1 this device of the mensuration of learning and memory function (diving tower method) is the case of 15cm * 30cm * 30cm; The shop is with the copper grid at the bottom of the case; Brass wire diameter is 2mm, and spacing is 5mm, and current intensity is controlled by an adjustable transformer; Pass to the 36V electric current, front left side is placed the valve rubber that high and a diameter is 4.5cm and is escaped the place of safety of clicking as mice in the case.Mice is put into the free movable 3min of case, be familiar with environment, pass to electric current at the bottom of the case then, jump onto platform during its normal reaction to hide noxious stimulation.To jump off the time that platform shocked by electricity for the 1st time be that the number of times that incubation period and 5min introskip lower platform are shocked by electricity is an errors to record from putting into, as school grade.Behind the 24h, Mus is placed on the platform, record is that the number of times that incubation period and 5min introskip lower platform are shocked by electricity is an errors from putting into to jumping off the time that platform shocked by electricity for the 1st time, as the memory achievement again.The result is through t check comparable group differences (down together).
5.2.2SOD, MAO, Na
+-K
+The ATPase Determination on content measure to finish back next day in memory function, takes off cervical vertebra and puts to death animal, on the ice platform, takes out little cerebral tissue fast and places the rinsing of ice normal saline; Remove bloodstain, filter paper is wiped away dried, after accurately weighing; Little cerebral tissue is immersed rapidly in 9 times of amount cold salines; Fully mill in ice bath with homogenizer, process 10% brain tissue homogenate, with 3000rmin
-1Centrifugal 10min, it is for use to get supernatant.Press the test kit description and measure SOD, MAO, Na
+-K
+-ATPase is active.
5.3 result:
5.3.1 the present invention is to the influence of model mice learning and memory function
Model group learning and memory of little mouse latent time obviously shortens, and learning and memory errors number showed increased relatively has significant difference (P＜0.01) with normal group; The high, medium and low dose groups of the present invention can obviously increase dementia mice learning and memory latent time, compares (P＜0.01) with model group, makes the learning and memory errors number obviously reduce (P＜0.01) simultaneously.
The influence of table 6 pair learning and memory of little mouse function (
n=10)
Annotate: compare with model group,
#P＜005,
*P＜0.01
5.3.2 the present invention is to model mice cerebral tissue SOD, MAO, Na
+-K
+The active influence of-ATPase
Model group SOD in Mice, Na
+-K
+-ATPase active obviously reduction, MAO activity increase, and with normal group significant difference (P＜0.01) are arranged relatively; The high, medium and low dose groups of the present invention can obviously raise senile dementia SOD in Mice, Na
+-K
+-ATPase is active, the activity of reduction MAO, with model group significant difference (P＜0.05, P＜0.01=is arranged relatively.
Table 7 pair mouse brain tissue SOD, MAO, Na
+-K
+-ATPase activity influence (
N=10)
Annotate: compare with model group,
#P＜0.05,
*P＜0.01
The specific embodiment
Embodiment 1: Radix Ginseng 220, Rhizoma Acori Graminei 190, Rhizoma Chuanxiong 220, Folium Ginkgo 190, Radix Rehmanniae Preparata 260, Radix Polygalae 110; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 8 times of water gaging distillations 9 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected.Volatile oil is processed clathrate with beta-cyclodextrin, drying under reduced pressure.Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 8 times of water gagings and decoct 2 times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered; Concentrating under reduced pressure (0.08MPa) to relative density 1.00-1.10 (20 ℃), adds ethanol and makes and contain alcohol amount and reach 70%, left standstill 24 hours, get supernatant; Reclaim ethanol and be concentrated into relative density 1.05 (20 ℃), left standstill 12 hours, get supernatant and filter, be concentrated into relative density 1.25; Drying under reduced pressure is ground into fine powder, with the beta-cyclodextrin clathrate mixing of microcrystalline Cellulose, volatile oil; Tabletting, film coating promptly gets tablet.
Embodiment 2: Radix Ginseng 280, Rhizoma Acori Graminei 220, Rhizoma Chuanxiong 280, Folium Ginkgo 220, Radix Rehmanniae Preparata 340, Radix Polygalae 130; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 12 times of water gaging distillations 2 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected.Volatile oil is processed clathrate with beta-cyclodextrin, drying under reduced pressure.Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 12 times of water gagings and decocted 2 hours, and merging filtrate and above-mentioned debris filter, and concentrating under reduced pressure is (0.08MPa) to relative density 1.05-1.10 (20 ℃); Add ethanol and make and contain alcohol amount and reach 40%, left standstill 30 hours, get supernatant; Reclaim ethanol and be concentrated into relative density 1.08 (20 ℃), left standstill 24 hours, get supernatant and filter; Be concentrated into relative density 1.25, drying under reduced pressure is ground into fine powder; With the beta-cyclodextrin clathrate mixing of microcrystalline Cellulose, volatile oil, encapsulated, promptly get hard capsule.
Embodiment 3: Radix Ginseng 190, Rhizoma Acori Graminei 310, Rhizoma Chuanxiong 190, Folium Ginkgo 310, Radix Rehmanniae Preparata 225, Radix Polygalae 190; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 9 times of water gaging distillations 8 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected.Volatile oil is processed clathrate with beta-cyclodextrin, drying under reduced pressure.Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 9 times of water gagings and decoct 3 times, and each 1 hour, merging filtrate and above-mentioned debris filtered; Concentrating under reduced pressure (0.08MPa) to relative density 1.00-1.05 (20 ℃), adds ethanol and makes and contain alcohol amount and reach 60%, left standstill 8 hours, get supernatant; Reclaim ethanol and be concentrated into relative density 1.07 (20 ℃), left standstill 30 hours, get supernatant and filter, be concentrated into relative density 1.25; Drying under reduced pressure is ground into fine powder, adds an amount of mannitol and dextrin; Mixing makes soft material with 60% ethanol, crosses 12 mesh sieves; Drying under reduced pressure, granulate adds cyclodextrin clathrate and 0.3% stevioside, and mixing promptly gets granule.
Embodiment 4: Radix Ginseng 310, Rhizoma Acori Graminei 280, Rhizoma Chuanxiong 310, Folium Ginkgo 280, Radix Rehmanniae Preparata 375, Radix Polygalae 170; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 11 times of water gaging distillations 7 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected.Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 11 times of water gagings and decocted 1.5 hours, and merging filtrate and above-mentioned debris filter, and concentrating under reduced pressure is (0.08MPa) to relative density 1.05-1.10 (20 ℃); Add ethanol and make and contain alcohol amount and reach 65%, left standstill 12 hours, get supernatant, reclaim ethanol and also be concentrated into relative density 1.08 (20 ℃); Left standstill 18 hours, and got supernatant and filter, be concentrated into relative density 1.25, drying under reduced pressure; Be ground into fine powder, adding Polyethylene Glycol substrate is an amount of, is heated to fusing, adds above-mentioned volatile oil again; Mixing splashes in the refrigerative liquid paraffin, processes drop pill.
Embodiment 5: Radix Ginseng 280, Rhizoma Acori Graminei 310, Rhizoma Chuanxiong 280, Folium Ginkgo 310, Radix Rehmanniae Preparata 340, Radix Polygalae 190; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 10 times of water gaging distillations 5 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected.Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 10 times of water gagings and decoct 3 times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered; Concentrating under reduced pressure (0.08MPa) to relative density 1.05-1.10 (20 ℃), adds ethanol and makes and contain alcohol amount and reach 55%, left standstill 20 hours, get supernatant; Reclaim ethanol and be concentrated into relative density 1.06 (20 ℃), left standstill 24 hours, get supernatant and filter, be concentrated into relative density 1.25; Drying under reduced pressure is ground into fine powder, adds an amount of soybean oil; With above-mentioned volatile oil mixing, process soft capsule again, promptly get.
Embodiment 6: Radix Ginseng 190g, Rhizoma Acori Graminei 220g, Rhizoma Chuanxiong 190g, Folium Ginkgo 220g, Radix Rehmanniae Preparata 225g, Radix Polygalae 130g; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 10 times of water gagings and decoct 2 times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered; Concentrating under reduced pressure (0.08MPa) to relative density 1.08-1.10 (20 ℃), adds ethanol and makes and contain alcohol amount and reach 50%, left standstill 24 hours, get supernatant; Reclaim ethanol and be concentrated into relative density 1.06 (20 ℃), left standstill 24 hours, get supernatant and filter, be concentrated into the thick paste of relative density 1.25; Rhizoma Chuanxiong, the Radix Angelicae Dahuricae are ground into fine powder, add above-mentioned thick paste, mixing, pill; Drying, polishing promptly gets.
Embodiment 7: Radix Ginseng 250g, Rhizoma Acori Graminei 250g, Rhizoma Chuanxiong 250g, Folium Ginkgo 250g, Radix Rehmanniae Preparata 300g, Radix Polygalae 150g; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 10 times of water gaging distillations 6 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 10 times of water gagings and decoct three times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered; 20 ℃ are evaporated to relative density 1.08-1.10, add ethanol and make and contain alcohol amount and reach 50%, leave standstill 24 hours, get supernatant; Reclaim ethanol and 20 ℃ and be concentrated into relative density 1.06, left standstill 24 hours, get supernatant and filter; Filtrating adds above-mentioned volatile oil, cyclamate 1.5g and potassium sorbate 2g, tween 80 20ml, ethyl hydroxybenzoate 0.3g, adds purified water, with the sodium hydroxide adjust pH of dilute hydrochloric acid or 40% to 4.0-6.0; Filter, sterilization promptly gets oral liquid.
Claims (12)
1. Chinese medicine composition of treating mental retardation and senile dementia; It is characterized in that according to the weight portion meter raw material of processing this Chinese medicine composition active component is: Radix Ginseng 190-310, Rhizoma Acori Graminei 190-310, Rhizoma Chuanxiong 190-310, Folium Ginkgo 190-310, Radix Rehmanniae Preparata 225-375, Radix Polygalae 110-190.
2. the Chinese medicine composition of treatment mental retardation as claimed in claim 1 and senile dementia; It is characterized in that according to the weight portion meter, process the said composition raw materials of effective components and be: Radix Ginseng 220-280, Rhizoma Acori Graminei 220-280, Rhizoma Chuanxiong 220-280, Folium Ginkgo 220-280, Radix Rehmanniae Preparata 260-340, Radix Polygalae 130-170.
3. the Chinese medicine composition of treatment mental retardation as claimed in claim 2 and senile dementia; It is characterized in that according to the weight portion meter, process the said composition raw materials of effective components and be: Radix Ginseng 250, Rhizoma Acori Graminei 250, Rhizoma Chuanxiong 250, Folium Ginkgo 250, Radix Rehmanniae Preparata 300, Radix Polygalae 150.
4. like the Chinese medicine composition of claim 1,2 or 3 described treatment mental retardation and senile dementia, it is characterized in that said Chinese medicine composition is pill, granule, hard capsule, soft capsule, tablet or oral liquid.
5. the Chinese medicine composition of treatment mental retardation as claimed in claim 4 and senile dementia is characterized in that said Chinese medicine composition is an oral liquid.
6. the oral liquid of the Chinese medicine composition of treatment mental retardation as claimed in claim 5 and senile dementia is characterized in that it prepares through following method: Radix Ginseng 250g, Rhizoma Acori Graminei 250g, Rhizoma Chuanxiong 250g, Folium Ginkgo 250g, Radix Rehmanniae Preparata 300g, Radix Polygalae 150g; Rhizoma Chuanxiong, Rhizoma Acori Graminei add 10 times of water gaging distillations 6 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 10 times of water gagings and decoct three times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered; Being evaporated to 20 ℃ of relative densities is 1.08-1.10, adds ethanol and makes and contain alcohol amount and reach 50%, leaves standstill 24 hours, gets supernatant; Reclaiming ethanol and being concentrated into 20 ℃ of relative densities is 1.06, leaves standstill 24 hours, gets supernatant and filters; Filtrating adds above-mentioned volatile oil, an amount of solubilizing agent, correctives, antiseptic, and the adjustment volume filters to 1000ml; Packing, sterilization promptly gets.
7. like the method for preparing of the Chinese medicine composition of described treatment mental retardation of the arbitrary claim of claim 1-5 and senile dementia, it is characterized in that: Rhizoma Chuanxiong, Rhizoma Acori Graminei two flavor distillating extracting oils; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae are carried out decocting; Add proper auxiliary materials, process preparation.
8. the method for preparing of the Chinese medicine composition of treatment mental retardation as claimed in claim 7 and senile dementia is characterized in that: Rhizoma Chuanxiong, Rhizoma Acori Graminei add 8-12 times of water gaging distillation 2-9 hour, and extraction volatile oil is subsequent use or enclose is dry, and debris device is in addition collected; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 8-12 times of water gaging and decoct 1-3 time, and each 1-2 hour, merging filtrate and above-mentioned debris; Filter, being evaporated to 20 ℃ of relative densities is 1.00-1.20, adds ethanol and makes and contain the alcohol amount and reach 40-70%; Left standstill 8-30 hour, and got supernatant, reclaiming ethanol and being concentrated into 20 ℃ of relative densities is 1.00-1.10; Left standstill 12-30 hour, and got supernatant filtration or concentrate drying, adding volatile oil and proper auxiliary materials, process preparation.
9. like the method for preparing of the Chinese medicine composition of claim 7 or 8 described treatment mental retardation and senile dementia, it is characterized in that: Rhizoma Chuanxiong, Rhizoma Acori Graminei add 10 times of water gagings distillations 6 hours, and it is subsequent use to extract volatile oil, and debris device is in addition collected; Radix Ginseng, Folium Ginkgo, Radix Rehmanniae Preparata, Radix Polygalae add 10 times of water gagings and decoct three times, and each 1.5 hours, merging filtrate and above-mentioned debris filtered; Being evaporated to 20 ℃ of relative densities is 1.08-1.10, adds ethanol and makes and contain alcohol amount and reach 50%, leaves standstill 24 hours, gets supernatant; Reclaiming ethanol and being concentrated into 20 ℃ of relative densities is 1.06, leaves standstill 24 hours, gets supernatant and filters, and filtrating adds above-mentioned volatile oil, an amount of solubilizing agent, correctives, antiseptic; Filter, packing, sterilization promptly gets oral liquid.
10. the discrimination method of the oral liquid of the Chinese medicine composition of treatment mental retardation as claimed in claim 6 and senile dementia is characterized in that this method comprises one or more of following method:
(1) thin layer of Rhizoma Acori Graminei is differentiated: get 8ml of the present invention, with extracted with diethyl ether 2 times, 20ml obtains ether layer at every turn, and evaporate to dryness, residue add ethyl acetate 1ml, make dissolving, as need testing solution; Other gets Rhizoma Acori Graminei control medicinal material 2g, the 25ml that adds diethyl ether, and supersound process 10min filters, the filtrating evaporate to dryness, residue adds ethyl acetate 1ml, makes dissolving, as control medicinal material solution; According to " each 5 μ l of above-mentioned 2 kinds of solution are drawn in the test of Chinese pharmacopoeia thin layer chromatography, and putting in the same 0.5%CMC-Na of containing solution respectively is on the silica gel g thin-layer plate of binding agent; With petroleum ether-ethyl acetate of 60-90 ℃=8: 2 was developing solvent, launched, and exhibition is apart from 8cm; Take out, dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show identical fluorescence speckle; Smoked clear with iodine vapor again to the speckle colour developing, inspect under the daylight, show identical sepia speckle;
(2) thin layer of Rhizoma Chuanxiong is differentiated: get 8ml of the present invention, with extracted with diethyl ether 2 times, 20ml obtains ether layer at every turn, and evaporate to dryness, residue add ethyl acetate 1ml, make dissolving, as need testing solution; Other gets Rhizoma Chuanxiong control medicinal material 2g, the 25ml that adds diethyl ether, and supersound process 10min filters, and gets the filtrating evaporate to dryness, and residue adds ethyl acetate 1ml makes dissolving, as control medicinal material solution; According to " Chinese pharmacopoeia thin layer chromatography test is drawn each 10 μ l of above-mentioned 2 kinds of solution and put respectively on same silica gel g thin-layer plate, is developing solvent with normal hexane-ethyl acetate=9: 1; Launch, exhibition is taken out apart from 8cm; Dry, put under the 365nm ultra-violet lamp and inspect, in the test sample chromatograph; With the corresponding position of control medicinal material chromatograph on, show identical fluorescence speckle;
(3) thin layer of Radix Ginseng is differentiated: get 2mL of the present invention, thin up is to 15ml, with water saturated n-butanol extraction 2 times, and 25mL at every turn; Merge butanol extraction liquid, n-butyl alcohol liquid is with 0.1%NaOH solution washing 2 times, 15mL at every turn; The saturated water washing of reuse n-butyl alcohol 2 times, each 20mL abandons or adopts water layer; N-butyl alcohol liquid is put evaporate to dryness in the water-bath, and residue adds methanol 1mL dissolving, as need testing solution; Other get the reference substance ginsenoside Rb1 add methanol process respectively contain 1mgmL-1 mixed solution as reference substance solution.According to " each 10 μ L of above-mentioned 2 kinds of solution are drawn in the test of Chinese pharmacopoeia thin layer chromatography, put respectively on same silica gel g thin-layer plate; With chloroform-ethyl acetate-methanol-water=15: 40: 22: 10 was developing solvent at 10 ℃ of lower floor's solution with held; Launch, take out airing; Spray is dried by the fire to clear spot in 105 ℃ with 10% ethanol solution of sulfuric acid; Put respectively under daylight and the 365nm ultra-violet lamp and inspect, in the test sample chromatograph, with control medicinal material chromatograph and the corresponding position of reference substance chromatograph on, show the speckle of same color;
(4) thin layer of Radix Polygalae is differentiated: get 10ml of the present invention, divide 2 extractions with ether, each 20ml merges ether solution, washes 3 times, each 30ml, and ether solution low temperature evaporate to dryness, residue add ethyl acetate 0.5ml makes dissolving, as need testing solution; Other gets Radix Polygalae control medicinal material 1.5g, adds water 50ml and decocts 1h, filters, and filtrating is divided 2 extractions with ether, and merge extractive liquid, is concentrated into 1ml, as control medicinal material solution; According to " Chinese pharmacopoeia thin layer chromatography test; Drawing each 4 μ l of above-mentioned 2 kinds of solution puts respectively on the silica gel g thin-layer plate of same usefulness 1% sodium hydroxide solution preparation; With cyclohexane extraction-chloroform-ethyl acetate-methanol=30: 15: 5: 0.5 was developing solvent, launches, and takes out; Dry, put under the 365nm ultra-violet lamp and inspect; In the test sample chromatograph, with the corresponding position of control medicinal material chromatograph on, show the fluorescence speckle of same color.
11. the content assaying method of the Chinese medicine composition of treatment mental retardation as claimed in claim 6 and senile dementia is characterized in that: use octadecylsilane chemically bonded silica to be filler; Methanol-1% glacial acetic acid=28: 72 is a mobile phase; The detection wavelength is 321nm; Theoretical cam curve is pressed the ferulic acid peak and is calculated, and should be not less than 2000; It is an amount of that precision takes by weighing the ferulic acid reference substance, adds methanol and process the solution that every ml contains 12ug, as reference substance solution; Accurate absorption 1ml of the present invention puts in the volumetric flask of 25ml, adds mobile phase and is diluted to scale, shakes up, as need testing solution; According to " the Chinese pharmacopoeia HPLC, accurate reference substance solution and each 5ul of need testing solution of drawing injects chromatograph of liquid respectively, measures, and promptly gets.
12. claim 1, the application of 2 or 3 described Chinese medicine compositions in preparation treatment mental retardation and medicine for senile dementia.
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| CN102526583B (en) * | 2012-03-12 | 2014-08-20 | 山西中医学院 | Chinese medicinal composition for treating senile dementia and preparation method and application thereof |
| CN103055014B (en) * | 2012-12-31 | 2014-09-17 | 青岛华仁技术孵化器有限公司 | Folium ginkgo health product for preventing senile dementia and preparation method thereof |
| CN103083558B (en) * | 2013-01-10 | 2015-07-22 | 山东凤凰制药股份有限公司 | Novel application of traditional Chinese medicine composition for treating cerebral infarction during acute stage and earlier restoration stage |
| CN105232972A (en) * | 2015-09-30 | 2016-01-13 | 郑梅 | Preparation for treating senile dementia and preparation method thereof |
| CN105920476B (en) * | 2016-04-13 | 2020-02-21 | 深圳市老年医学研究所 | Traditional Chinese medicine composition for preventing and treating Alzheimer disease and preparation method thereof |
| CN106491680B (en) * | 2016-12-14 | 2020-05-22 | 四川新绿色药业科技发展有限公司 | A Chinese medicinal composition for preventing or treating senile dementia, and its preparation method |
| CN114288353A (en) * | 2022-01-19 | 2022-04-08 | 宝莱福健康科技研究(中山)有限公司 | Preparation method and application of composition for preventing and treating Alzheimer's disease |
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