CN103316151B - Combined drug of rhodiola rosea extract, medlar extract and sea-buckthorn fresh pulp powder extract as well al preparation and application thereof - Google Patents
Combined drug of rhodiola rosea extract, medlar extract and sea-buckthorn fresh pulp powder extract as well al preparation and application thereof Download PDFInfo
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Abstract
The invention provides a combined drug of rhodiola rosea extract, medlar extract and sea-buckthorn fresh pulp powder extract as well al preparation and an application thereof, relates to an extraction and combination method and a preparation method of a medicinal preparation and in particular relates to an extraction and combination method of rhodiola rosea extract, medlar extract and sea-buckthorn fresh pulp powder extract and a preparation of rhodiola rosea extract, medlar extract and sea-buckthorn fresh pulp powder extract. The combined drug is composed of rhodiola rosea extract, medlar extract and sea-buckthorn fresh pulp powder extract, wherein weight ratio of rhodiola rosea extract to medlar extract to sea-buckthorn fresh pulp powder extract is (322-644):(85-340):(2.41-9.66). Salidroside content in rhodiola rosea extract is not less than 1.2%, lycium barbarum polysaccharide content in medlar extract, metered by glucose, is not less than 3.5%, and total flavone content in sea-buckthorn fresh pulp powder extract, metered by rutin, is not less than 20.0%. Each dose unit of the prepared combined drug of rhodiola rosea extract, medlar extract and sea-buckthorn fresh pulp powder extract contains 4.32-8.64mg of salidroside, 3.25-12.99mg of lycium barbarum polysaccharide and 0.483-1.931mg of total flavone.
Description
Technical field
The composition of medicine of a kind of Radix Rhodiolae extract of the present invention, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts and preparation thereof, application relate to a kind of extraction of pharmaceutical preparation, combined method and preparation method, more particularly, the extraction of a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, combined method and preparation thereof is related to.
Background technology
Make commodity by Radix Rhodiolae, Fructus Lycii and fresh syrup of Fructrs Hippophae through suitable process and have XINNAOXIN JIAONANG etc., its quality standard is WS-10628 (ZD-0628)-2002; Function is supplementing QI and nourishing YIN, blood circulation promoting and blood stasis dispelling; For deficiency of both vital energy and Yin, the dizziness caused by stagnation of blood stasis, headache, cardiopalmus, asthma, weak; The erythrocytosis that anoxia causes
.
Radix Rhodiolae system Crassulaceae Radix Rhodiolae belongs to the dry root welding technology of Rhodiola crenulata (Hook.f.Thoms) H.Ohba.Successively isolate from rhodiola plant and add multiple chemical substance, comprise flavonoid, saponins, amino acids, phenols, Coumarins, terpenoid, mineral, enzyme, volatile oil and other some common macromolecular substances.Pharmaceutical research shows that its primary pharmacological activity composition is rhodioside and tyrosol etc. thereof.
Fructus Lycii is the dry mature fruit of plant of Solanaceae lycium barbarum Lycium barbarum L..It is clinical the most frequently used traditional tonic medicine, containing compositions such as a large amount of saccharide, aminoacid and betanins in Fructus Lycii, the polysaccharide Fructus Lycii that test proves in saccharide can improve the lymhocyte transformation rate of laboratory animal and macrophage phagocytic merit marrow, lycium barbarum polysaccharide (LBP) have remarkable synergism to the spleen beautiful jade bar cell transformation function that T cell mitogenic agent ConA induce, and the antibody response caused dependent cells antigen SRBC reacts and also has facilitation.
Main containing flavone compound and triterpene, steroid compound in fresh syrup of Fructrs Hippophae, wherein flavone compound is isorhamnetin, isorhamnetin-3-B-D-glucoside, isorhamnetin-3-p-rutinoside and Quercetin etc., there is anti-heart failure effect, also have antiinflammatory, anti-allergic effects, have obvious curative effect to digestive system and blood system.
XINNAOXIN JIAONANG production technology is more extensive, because Chinese crude drug is by the impact of the environment such as the place of production, implantation methods and weather, the effective ingredient often criticizing medical material may be inconsistent, the medical material amount of this direct regulation makes capsule with simple technique, do not quantize index composition, dose-effect relationship is unclear, causes the quality often criticizing product inconsistent, and fresh syrup of Fructrs Hippophae has been gathered seasonality and not easily preserving, thus affect pharmaceutical effectiveness.
Summary of the invention
The present invention seeks to provide the composition of medicine of a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts and preparation thereof, application for above-mentioned weak point, is a kind of supplementing QI and nourishing YIN, medicine of blood circulation promoting and blood stasis dispelling and preparation method thereof, application.1, by the effective site research in Radix Rhodiolae, Fructus Lycii, fresh syrup of Fructrs Hippophae medical material, 70% ethanol refluxing process is adopted to be extracted rhodioside constituents in Radix Rhodiolae; Decoction and alcohol sedimentation technique is extracted Fructus Lycii crude polysaccharides active component in Fructus Lycii; Because fresh syrup of Fructrs Hippophae has been gathered seasonality, after taking fresh syrup of Fructrs Hippophae to cross 80 mesh sieves, concentrate drying pulverizes the method for preserving, solving the problem that fresh syrup of Fructrs Hippophae is not easily preserved, providing crude drug for producing again throughout the year, and the main pharmacodynamics position flavonoid with 70% ethanol extraction in fresh syrup of Fructrs Hippophae.2, medical material through above-mentioned extract for effective ingredient after make extract, formulated extract Testing index simultaneously, and worked out quality standard.3, through pharmacodynamics orthogonal experiment, show that the amount (that is to say the amount of rhodioside) of Radix Rhodiolae extract in the drug effect of each prescription group is principal element, the amount of fresh syrup of Fructrs Hippophae powder extracts is secondary cause.Finally by the optimal proportion determining each extract after the test of pesticide effectiveness, so both improve the curative effect of medicine, ensure again often to criticize the basically identical property of curative effect of finished product simultaneously.Through pharmacological experiment, the preparation adopting the present invention to make has exceeded the drug effect of XINNAOXIN JIAONANG.
The optimum proportioning that the invention provides a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts combines, and combines the pharmaceutical preparation of preparation thus.
Composition of medicine of a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts and preparation method thereof, to apply be take following scheme to realize:
The composition of medicine of a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, it is characterized in that, be made up of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts weight proportion are 322-644:85-340:2.41-9.66.
The composition of medicine of described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, it is characterized in that, must not 1.2% be less than containing rhodioside in Radix Rhodiolae extract, must not 3.5% be less than containing lycium barbarum polysaccharide with glucose meter in Fructus Lycii extract, in fresh syrup of Fructrs Hippophae powder extracts, 20 .0% must not be less than containing total flavones in rutin.
The composition of medicine of described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, it is characterized in that, after Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts make composition of medicine in each minimum dose unit (i.e. every a slice, grain, bag etc.) containing rhodioside, lycium barbarum polysaccharide, total flavones amount be respectively following weight scope: 4.32-8.64(mg), 3.25-12.99(mg), 0.483-1.931(mg), instructions of taking is oral, 2 times on the one, each 2 minimum dose units.
Described a kind of Radix Rhodiolae extract, the composition of medicine of Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, it is characterized in that, Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts make each minimum dose unit (i.e. every a slice after composition of medicine, grain, bag etc.) in containing rhodioside, lycium barbarum polysaccharide, the amount of total flavones is respectively in following weight combination any one group: 4.32:6.49:1.931, 8.64:3.25:1.931, 8.64:6.49:0.483, 8.64:12.99:0.965, instructions of taking is oral, 2 times on the one, each 2 minimum dose units.
The composition of medicine preparation method of described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, is characterized in that,
(1) Radix Rhodiolae extract technological process: after selecting clean, fragmentation, weigh, put in extraction pot, add 70% ethanol 10 times amount, 8 times amount respectively, reflux, extract, twice.3 hours first times, second time 2 hours, merges twice reflux extracting liquid, reclaims after ethanol, is concentrated into thick paste, dry, be ground into fine powder and namely obtain extract.
(2) Fructus Lycii extract technological process: through selecting only, weighing, putting in extraction pot, decoct with water twice.First time adds water 10 times amount, and second time adds water 8 times amount, and the time is respectively 2 hours, merges twice decocting liquid, filters, and filtrate concentrates, and with 80% ethanol precipitate with ethanol, centrifugal filtering, gets solid content drying, is ground into fine powder and namely obtains extract.
(3) fresh syrup of Fructrs Hippophae powder extracts technological process: fresh syrup of Fructrs Hippophae crosses concentrate drying after 80 mesh sieves, is ground into coarse powder, gets coarse powder and add 70% ethanol 10 times amount, 8 times amount respectively, reflux, extract, twice.3 hours first times, second time 2 hours, merges twice reflux extracting liquid, reclaims after ethanol, is concentrated into thick paste, dry, be ground into fine powder and namely obtain extract.
(4) said extracted thing is after assay, takes Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts by weight ratio respectively, make preparation after adding pharmaceutic adjuvant by preparation process according to content data.
Described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts composition of medicine, is characterized in that, it is characterized in that its preparation is oral formulations.
The composition of medicine of described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, is characterized in that the pharmaceutic adjuvant containing oral formulations.
The composition of medicine of described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, is characterized in that oral formulations comprises hard capsule, soft capsule, granule, tablet.
The composition of medicine of described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts treats deficiency of both vital energy and Yin preparing, dizziness caused by stagnation of blood stasis, headache, cardiopalmus, asthma, weak, apply in the medicine of the erythrocytosis that anoxia causes.
The composition of medicine of described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts is applied to treatment deficiency of both vital energy and Yin, the dizziness caused by stagnation of blood stasis, headache, cardiopalmus, and asthma is weak; The erythrocytosis that anoxia causes; Treatment, prevention myocardial infarction and cerebral infarction.
The amount of formulation effect relationship that the present invention makes is analysed clearly, definite ingredients, and the quality of the pharmaceutical preparations is stablized controlled, and curative effect is better, can meet clinical higher level demand.
The composition of medicine of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts and preparation thereof, an application are a kind of supplementing QI and nourishing YIN, medicine of blood circulation promoting and blood stasis dispelling and preparation method thereof, application.1, by the effective site research in Radix Rhodiolae, Fructus Lycii, fresh syrup of Fructrs Hippophae medical material, 70% ethanol refluxing process is adopted to be extracted rhodioside constituents in Radix Rhodiolae; Decoction and alcohol sedimentation technique is extracted Fructus Lycii crude polysaccharides active component in Fructus Lycii; Because fresh syrup of Fructrs Hippophae has been gathered seasonality, after taking fresh syrup of Fructrs Hippophae to cross 80 mesh sieves, concentrate drying pulverizes the method for preserving, solving the problem that fresh syrup of Fructrs Hippophae is not easily preserved, providing crude drug for producing again throughout the year, and the main pharmacodynamics position flavonoid with 70% ethanol extraction in fresh syrup of Fructrs Hippophae.2, medical material through above-mentioned extract for effective ingredient after make extract, formulated extract Testing index simultaneously, and worked out quality standard.3, through pharmacodynamics orthogonal experiment, show that the amount (that is to say the amount of rhodioside) of Radix Rhodiolae extract in the drug effect of each prescription group is principal element, the amount of fresh syrup of Fructrs Hippophae powder extracts is secondary cause.Finally by the optimal proportion determining each extract after the test of pesticide effectiveness, so both improve the curative effect of medicine, ensure again often to criticize the basically identical property of curative effect of finished product simultaneously.Through pharmacological experiment, the preparation adopting the present invention to make has exceeded the drug effect of XINNAOXIN JIAONANG.
Detailed description of the invention
By the following examples, in conjunction with results of pharmacodynamic test, further illustrate the present invention, following embodiment does not only limit the present invention for illustration of the present invention.
A composition of medicine preparation method for Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, its method is as follows:
(1) Radix Rhodiolae extract technological process: after selecting clean, fragmentation, weigh, put in extraction pot, add 70% ethanol 10 times amount, 8 times amount respectively, reflux, extract, twice, 3 hours first times, second time 2 hours, merges twice reflux extracting liquid, after reclaiming ethanol, be concentrated into thick paste, dry, be ground into fine powder and namely obtain Radix Rhodiolae extract;
(2) Fructus Lycii extract technological process: through selecting only, weighing, putting in extraction pot, decoct with water twice, add water 10 times amount for the first time, and add water 8 times amount for the second time, and the time is respectively 2 hours, merge twice decocting liquid, filter, filtrate concentrates, with 80% ethanol precipitate with ethanol, centrifugal filtering, gets solid content drying, is ground into fine powder and namely obtains Fructus Lycii extract;
(3) fresh syrup of Fructrs Hippophae powder extracts technological process: fresh syrup of Fructrs Hippophae crosses concentrate drying after 80 mesh sieves, be ground into coarse powder, get coarse powder and add 70% ethanol 10 times amount, 8 times amount respectively, reflux, extract, twice, 3 hours first times, second time 2 hours, merge twice reflux extracting liquid, reclaim after ethanol, be concentrated into thick paste, dry, be ground into fine powder and namely obtain fresh syrup of Fructrs Hippophae powder extracts;
(4) said extracted thing is after assay, takes Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts by weight ratio respectively, make preparation after adding pharmaceutic adjuvant by preparation process according to content data.
Embodiment 1
Prescription (prescription 7):
Radix Rhodiolae extract (containing rhodioside 8.64mg × 1000) 644g
Fructus Lycii extract (containing lycium barbarum polysaccharide 3.25mg × 1000) 85g
Fresh syrup of Fructrs Hippophae powder extracts (containing total flavones 1.931mg × 1000) 9.66g
Method for making: take Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts by prescription, add appropriate amount of auxiliary materials mix homogeneously, obtain 1000 with capsule fill.
Embodiment 2
Prescription (prescription 7):
Radix Rhodiolae extract (containing rhodioside 8.64mg × 1000) 644g
Fructus Lycii extract (containing lycium barbarum polysaccharide 3.25mg × 1000) 85g
Fresh syrup of Fructrs Hippophae powder extracts (containing total flavones 1.931mg × 1000) 9.66g
Cera Flava 45g
Gelatin is appropriate
Glycerol is appropriate
Paraffin oil is appropriate
Method for making: get Cera Flava, slight fever makes it dissolve, and add Radix Rhodiolae extract, Fructus Lycii extract, fresh syrup of Fructrs Hippophae powder extracts and paraffin wet goods proper auxiliary materials, stir evenly, colloid mill grinds well, and decompression degasification, medicinal liquid is pressed into soft capsule 1000.
Embodiment 3
Prescription (prescription 7):
Radix Rhodiolae extract (containing rhodioside 8.64mg × 1000) 644g
Fructus Lycii extract (containing lycium barbarum polysaccharide 3.25mg × 1000) 85g
Fresh syrup of Fructrs Hippophae powder extracts (containing total flavones 1.931mg × 1000) 9.66g
Sucrose 600g
Starch 350g
Making beating starch 60g
Essence 2 ml
Method for making: take the Radix Rhodiolae extract of recipe quantity, Fructus Lycii extract, fresh syrup of Fructrs Hippophae powder extracts, sugarcane sugar and starch, pulverized 100 mesh sieves, and fully mixed half an hour; Making beating starch is washed into 10% starch slurry, lets cool for subsequent use.Starch slurry is joined in the material mixed under stirring, make soft material, cross 14 mesh sieves, the wet grain of system.Wet grain, in 55 DEG C of dryings 3 hours, sprays into essence after granulate, and place half an hour, subpackage obtains 1000 bags.
Embodiment 4
Prescription (prescription 7):
Radix Rhodiolae extract (containing rhodioside 8.64mg × 1000) 644g
Fructus Lycii extract (containing lycium barbarum polysaccharide 3.25mg × 1000) 85g
Fresh syrup of Fructrs Hippophae powder extracts (containing total flavones 1.931mg × 1000) 9.66g
Starch 80g
Microcrystalline Cellulose 85g
Carboxymethyl starch sodium 70g
Magnesium stearate 3g
Polyvidone
k30(PVP
k30) appropriate
Method for making: starch, microcrystalline Cellulose, carboxymethylstach sodium, magnesium stearate cross 80 mesh sieves respectively; Get the solution that PVP K30 anhydrous alcohol solution makes 5%, mix for subsequent use.Take Radix Rhodiolae extract, Fructus Lycii extract, fresh syrup of Fructrs Hippophae powder extracts, starch, microcrystalline Cellulose, carboxymethyl starch sodium by recipe quantity, mix homogeneously, add 5% PVP K30 liquid and make soft material, cross 20 mesh sieves, the wet grain of system, wet grain is put after 50 DEG C of dryings, crosses 18 mesh sieves, add magnesium stearate, after mixing, tabletting, film coating, make 1000, obtain tablet.
Embodiment 5
Prescription (prescription 5):
Radix Rhodiolae extract (containing rhodioside 4.32mg × 1000) 322g
Fructus Lycii extract (containing lycium barbarum polysaccharide 6.49mg × 1000) 170g
Fresh syrup of Fructrs Hippophae powder extracts (containing total flavones 1.931mg × 1000) 9.66g
Starch 60g
Microcrystalline Cellulose 50 g
Carboxymethyl starch sodium 70g
Magnesium stearate 2g
Polyvidone
k30(PVP
k30) appropriate
Method for making: starch, microcrystalline Cellulose, carboxymethylstach sodium, magnesium stearate cross 80 mesh sieves respectively; Get the solution that PVP K30 anhydrous alcohol solution makes 5%, mix for subsequent use.Take Radix Rhodiolae extract, Fructus Lycii extract, fresh syrup of Fructrs Hippophae powder extracts, starch, microcrystalline Cellulose, carboxymethyl starch sodium by recipe quantity, mix homogeneously, add 5% PVP K30 liquid and make soft material, cross 20 mesh sieves, the wet grain of system, wet grain is put after 50 DEG C of dryings, crosses 18 mesh sieves, add magnesium stearate, after mixing, tabletting, film coating, make 1000, obtain tablet.
Embodiment 6
Prescription (prescription 8):
Radix Rhodiolae extract (containing rhodioside 8.64mg × 1000) 644g
Fructus Lycii extract (containing lycium barbarum polysaccharide 6.49mg × 1000) 170g
Fresh syrup of Fructrs Hippophae powder extracts (containing total flavones 0.483mg × 1000) 2.41g
Starch 70g
Microcrystalline Cellulose 65g
Carboxymethyl starch sodium 70g
Magnesium stearate 3g
Polyvidone
k30(PVP
k30) appropriate
Method for making: starch, microcrystalline Cellulose, carboxymethylstach sodium, magnesium stearate cross 80 mesh sieves respectively; Get the solution that PVP K30 anhydrous alcohol solution makes 5%, mix for subsequent use.Take Radix Rhodiolae extract, Fructus Lycii extract, fresh syrup of Fructrs Hippophae powder extracts, starch, microcrystalline Cellulose, carboxymethyl starch sodium by recipe quantity, mix homogeneously, add 5% PVP K30 liquid and make soft material, cross 20 mesh sieves, the wet grain of system, wet grain is put after 50 DEG C of dryings, crosses 18 mesh sieves, add magnesium stearate, after mixing, tabletting, film coating, make 1000, obtain tablet.
Embodiment 7
Prescription (prescription 9):
Radix Rhodiolae extract (containing rhodioside 8.64mg × 1000) 644g
Fructus Lycii extract (containing lycium barbarum polysaccharide 12.99mg × 1000) 340g
Fresh syrup of Fructrs Hippophae powder extracts (containing total flavones 0.965mg × 1000) 4.83g
Starch 80g
Microcrystalline Cellulose 85g
Carboxymethyl starch sodium 70g
Magnesium stearate 3g
Polyvidone
k30(PVP
k30) appropriate
Method for making: starch, microcrystalline Cellulose, carboxymethylstach sodium, magnesium stearate cross 80 mesh sieves respectively; Get the solution that PVP K30 anhydrous alcohol solution makes 5%, mix for subsequent use.Take Radix Rhodiolae extract, Fructus Lycii extract, fresh syrup of Fructrs Hippophae powder extracts, starch, microcrystalline Cellulose, carboxymethyl starch sodium by recipe quantity, mix homogeneously, add 5% PVP K30 liquid and make soft material, cross 20 mesh sieves, the wet grain of system, wet grain is put after 50 DEG C of dryings, crosses 18 mesh sieves, add magnesium stearate, after mixing, tabletting, film coating, make 1000, obtain tablet.
radix Rhodiolae extract quality standard
differentiate:test according to thin layer chromatography, draw the reference substance solution under (assay) item and each 10 μ l of need testing solution, put respectively on same silica gel G lamellae, with lower floor's solution of chloroform-methanol-acetone-water (6:3:1:1) for developing solvent, launch, exhibition, apart from 18cm, is taken out, dry, put in iodine vapor smoked.In test sample chromatograph, on the position corresponding to reference substance chromatograph, the speckle of aobvious same color.
moisture:get this product 2 ~ 5g, put 100 ~ 105 DEG C and be dried to constant weight (double weight differential is no more than 5mg), should 10.0% be crossed.
assay:chromatographic condition and system suitability with octadecyl silicon protective embankment bonded silica gel for filler; With methanol-water (15:85) for mobile phase; Determined wavelength is 275nm.Number of theoretical plate calculates should be not less than 2000 by rhodioside peak.
The preparation of reference substance solution gets rhodioside reference substance in right amount, accurately weighed, adds methanol and makes the solution of every l m l containing 0.5mg, to obtain final product.
This product 0.2g is got in the preparation of need testing solution, accurately weighed, and put in tool plug conical flask, precision adds methanol 10ml, close plug, weighed weight, supersound process 30 minutes, lets cool, more weighed weight, supplies the weight of less loss with methanol, shake up, filter, get subsequent filtrate, to obtain final product.
Algoscopy is accurate respectively draws reference substance solution and each 10 μ l of need testing solution, injection liquid chromatography, measures, to obtain final product.
This product is pressed dry product and is calculated, containing rhodioside (C
14h
20o
7) must not 1.2% be less than.
fructus Lycii extract quality standard
differentiate:take extract and be about 0.25g, add water 35ml, and heated and boiled 15 minutes, lets cool, and filters, and filtrate is extracted with ethyl acetate 15ml jolting, divides and get acetic acid ethyl fluid, be concentrated into lml, as need testing solution.Separately get Fructus Lycii control medicinal material 0.5g, be made in the same way of control medicinal material solution.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, with ethyl acetate-chloroform-formic acid, (3:2:1 >, for developing solvent, launches, and takes out, dry, inspect under putting ultra-violet lamp (365nm).In test sample chromatograph, on the position corresponding to control medicinal material chromatograph, the fluorescence speckle of aobvious same color.
moisture:get this product 2 ~ 5g, put 100 ~ 105 DEG C and be dried to constant weight (double weight differential is no more than 5mg), should 10.0% be crossed.
assay:anhydrous glucose reference substance 25mg is got in the preparation of reference substance solution, accurately weighed, puts in 250ml measuring bottle, and add water appropriate dissolving, is diluted to scale, shakes up, and obtains (containing anhydrous glucose 0.lmg in every lml).
The preparation precision of standard curve measures reference substance solution 0.2ml, 0.4ml, 0.6ml, 0.8 m l, 1.0 m l, put in tool plug test tube respectively, add water respectively and mend to 2. 0ml, each precision adds 5% phenol solution lml, shake up, rapid precision adds sulphuric acid 5ml, shakes up, place 10 minutes, put in 40 DEG C of water-baths and be incubated 15 minutes, take out, be cooled to room temperature rapidly, with corresponding reagent for blank, according to ultraviolet visible spectrophotometry, measuring absorbance at the wavelength place of 490nm, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve.
Algoscopy is got this product and is about 0.25g, accurately weighed, and add diethyl ether 100ml, reflux 1 hour, leaves standstill, lets cool, carefully discard ether solution, and residue puts water-bath Back stroke ether to the greatest extent.Add 80 % ethanol 100 m l, reflux 1 hour, filters while hot, and filtering residue and the hot 80% ethanol 30ml gradation of filter are washed, and filtering residue is put in flask together with filter paper, and add water 150ml, reflux 2 hours.Filter while hot, with a small amount of hot wash filter, merging filtrate and washing liquid, let cool, move in 250ml measuring bottle, be diluted with water to scale, shake up, precision measures l m l, puts in tool plug test tube, add water 1.0ml, the method under the preparation of sighting target directrix curve, from " each precision adds 5% phenol solution 1ml ", measure absorbance in accordance with the law, read the weight (mg) containing glucose in need testing solution from standard curve, calculate, to obtain final product.This product is pressed dry product and is calculated, containing lycium barbarum polysaccharide with glucose (C
6h
12o
6) meter, must not 3.5% be less than.
fresh syrup of Fructrs Hippophae powder extracts quality standard
differentiate:get this product powder and be about 0.2g, put in tool plug conical flask, add ethanol 50ml, reflux 1 hour, lets cool filtration.Measure subsequent filtrate 25m l, put in tool plug conical flask, add hydrochloric acid 3. 5ml, heating hydrolysis 1 hour in 75 DEG C of water-baths, cool immediately, be transferred in 50ml measuring bottle, use ethanol in proper amount washing container, washing liquid is incorporated in same measuring bottle, add ethanol to scale, shake up, filter, get subsequent filtrate 30ml, be concentrated into about 5m l, add water 25ml, 2 times are extracted, each 20ml, combined ethyl acetate liquid by ethyl acetate, evaporate to dryness, residue adds methanol lml makes dissolving, as need testing solution.Separately get isorhamnetin reference substance, Quercetin reference substance, add methanol and make every lml respectively containing the mixed solution of lmg, product solution in contrast.Test according to thin layer chromatography, draw each 2 μ l of above-mentioned two kinds of solution, put respectively and contain on silica gel G lamellae prepared by 3% sodium acetate solution in same, with toluene-ethyl acetate-formic acid (5:2: 1) for developing solvent, launch, take out, dry, spray, with aluminum chloride test solution, is inspected under putting ultra-violet lamp (3 65nm).In test sample chromatograph, on the position corresponding to reference substance chromatograph, the fluorescence speckle of aobvious same color.
moisture:get this product 2 ~ 5g, put 100 ~ 105 DEG C and be dried to constant weight (double weight differential is no more than 5mg), should 10.0% be crossed.
assay:control substance of Rutin 20mg is got in the preparation of reference substance solution, accurately weighed, puts in 50ml measuring bottle, adds 60% appropriate amount of ethanol, puts slight fever in water-bath and makes dissolving, let cool, and adds 60% ethanol to scale, shakes up.Precision measures 25ml, puts in 50ml measuring bottle, is diluted with water to scale, shake up, and obtains (every lml is containing rutin 0.2mg).
The preparation precision of standard curve measures reference substance solution l m l, 2 m l, 3 m l, 4ml, 5ml, 6ml, put in 25ml measuring bottle respectively, respectively add 30% ethanol to 6.0ml, add 5% sodium nitrite solution lml, mixing, places 6 minutes, then adds 10% aluminum nitrate solution lml, shake hook, place 6 minutes.Hydro-oxidation sodium test solution 10ml, then add 30% ethanol to scale, shake up, place 15 minutes, be blank with corresponding reagent, according to ultraviolet visible spectrophotometry, measure absorbance at the wavelength place of 500nm, take absorbance as vertical coordinate, concentration is abscissa, drawing standard curve.
Algoscopy gets this product about 0.2 g, accurately weighed, adds 60 % ethanol 30ml, reflux 2 hours, lets cool, and filters, residue adds 60% ethanol 25ml more respectively, reflux 2 times, each 1 hour, filter, merging filtrate, puts in 100ml measuring bottle, residue 60% washing with alcohol, washing liquid is incorporated in same measuring bottle, with 60% ethanol dilution to scale, shakes up.Precision measures 25ml, puts in 50ml measuring bottle, adds water to scale, shake up, as need testing solution.Precision measures need testing solution 3ml, puts in 25ml measuring bottle, adds 30% ethanol to 6ml, method under sighting target directrix curve preparation, from " adding sodium nitrite solution lml ", measure absorbance in accordance with the law, get need testing solution 3ml simultaneously, except not hydro-oxidation sodium test solution, all the other the same operations, as blank, read the weight (mg) containing rutin in need testing solution from standard curve, calculate, to obtain final product.
This product is pressed dry product and is calculated, containing total flavones with rutin (C
27h
30o
16) meter, 20 .0% must not be less than.
Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts composition of medicine effect experiment are studied and quadrature analysis
1. sketch:this experiment adopts rat arteriovenous shunt thrombosis formation model, measures wet weight of thrombus and dry weight; ADP induces rabbit platelet aggregation, measures maximum platelet aggregation rate.Result shows, in arteriovenous shunt thrombosis is formed, Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts composition of medicine (hereinafter referred to as composition of medicine) prescription 7 extremely significantly can suppress the formation (P<0.01) of wet weight of thrombus, and its inhibitory action is better than XINNAOXIN JIAONANG group and all the other prescription groups; Composition of medicine prescription 7 and XINNAOXIN JIAONANG group all have stronger inhibitory action to thrombosis dry weight, and both effects are similar; Induce in rabbit platelet aggregation experiment at ADP, composition of medicine prescription 7 can extremely remarkable anticoagulant rate (P<0.01), and its effect is better than compound Salviae Miltiorrhizae group, XINNAOXIN JIAONANG group and all the other prescription groups.Adopt direct-vision method to align friendship interpretation of result, in preliminary discovery composition of medicine prescription, the amount of Radix Rhodiolae extract plays a major role in the test of pesticide effectiveness, and fresh syrup of Fructrs Hippophae powder extracts plays a part secondary.
experiment purpose
According to the requirement of new drug preclinical study guideline, the Pharmacodynamics effect of research XINNAOXIN JIAONANG and composition of medicine prescription group, for clinical practice provides experimental basis.
3. experiment material
3.1 laboratory animal
Cleaning grade SD rat, male and female half and half, body weight 250 ~ 280g, Suzhou Industrial Park Ai Ermaite Science and Technology Ltd. provides, and the animal quality quality certification number: SCXK(revives) 2009-0001
Male rabbit, 1.8 ~ 2.0kg, Qinglongshan animal cultivation field, Jiangning county, the animal quality quality certification number: SCXK(revives) 2012-0008
3.2 medicines and reagent
XINNAOXIN JIAONANG: S&P Pharmaceutical Co., Ltd., lot number: 120803.Specification: 0.5g.Before use grind into powder and with 0.5% the even suspendible of CMC-Na.
Radix Rhodiolae extract: lot number: 20121221
Fructus Lycii extract: lot number: 20130111
Fresh syrup of Fructrs Hippophae powder extracts: lot number: 20130112
the list of composition of medicine people consumption experimental program
Each prescription be mixed in proportion rear grinding before use and with 0.5% the even suspendible of CMC-Na.
corresponding each index components content in the minimum dose unit of each prescription group and XINNAOXIN JIAONANG
FUFANG DANSHEN PIAN: Guangdong Jian Yile pharmaceutical Co. Ltd, lot number: 120909.Before use grind into powder and with 0.5% the even suspendible of CMC-Na.
Chloral hydrate: Chemical Reagent Co., Ltd., Sinopharm Group, lot number: 30037517, is mixed with 0.9% sodium chloride injection the settled solution that concentration is 3% during use.
ADP：Biosharp。Lot number: Amresco0160.The settled solution of desired concn is become before use with normal saline.
0.9% sodium chloride injection: Jiangsu Pengyao Pharmaceutical Co., Ltd., lot number: 0909242
3.3 experimental apparatus
Electronic balance: Beijing Sai Duolisi instrument system company limited product.
Table-type low-speed centrifuge: Shanghai Medical apparatus (group) company limited's operating theater instruments field
Baking oven: Shanghai Fuma Experiment Equipment Co., Ltd.
PAPER-I type platelet aggregation and blood clotting analyzer: Beijing Steellex Scientific Instrument Company's product
4. experimental technique and result
The impact that 4.1 XINNAOXIN JIAONANG prescriptions are formed rat arteriovenous shunt thrombosis
Cleaning grade rat is divided into 11 groups at random, often organizes 8, male and female half and half.Model control group, positive control XINNAOXIN JIAONANG 200 mg/kg, prescription group 1 ~ 9 dosage is respectively 99.36,134.33,204.26,164.73,200.66,265.76,295.46,326.56,395.53mg/kg, administration volume is 1ml/100g.Gavage every day (ig) is administered once, continuous gavage 7 days, 0.5h after last administration, rat 3% chloral hydrate (300mg/kg) ip is anaesthetized, cervical region median incision, be separated right carotid and left external jugular vein, 7 trumpeter's art silk threads of a long 6cm are put in the stage casing of three sections of polyethylene tubes, normal saline solution is full of polyethylene tube, right carotid is inserted in its one end, the other end inserts left external jugular vein, opens bulldog clamp, and blood returns left external jugular vein from right common carotid artery through polyethylene tube.Open blood flow 15 min, middle Herba Clinopodii, take out rapidly silk thread and weigh, gross weight deducts silk thread weight, is wet weight of thrombus.Then put into the roasting 20min of baking box (60 DEG C), weigh immediately, this weight deducts silk thread weight and is thrombosis dry weight.By following formulae discovery suppression ratio:
The results are shown in Table 1.
Note: data statistical approach: total data with
represent, statistical method is that between two groups, t inspection is compared, and P<0.05 represents that between two groups, data have significant difference, and P<0.01 represents that between two groups, data have pole significant difference.
From table 1, XINNAOXIN JIAONANG group and each prescription group all can alleviate weight in wet base and the dry weight of thrombosis.Compared with model control group, XINNAOXIN JIAONANG group and prescription 5,7,8,9 groups significantly can reduce wet weight of thrombus (P<0.05), prescription 7,8,9 extremely significantly can reduce wet weight of thrombus (P<0.01), prescription 1,2,3,4,6 groups does not have significant difference (P>0.05) compared with model control group, compared with XINNAOXIN JIAONANG group, prescription 7 significantly can reduce wet weight of thrombus (P<0.05); Compared with model control group, XINNAOXIN JIAONANG group, prescription 7 all significantly can suppress thrombosis dry weight (P<0.05), prescription 1,2,3,4,5,6,8,9 groups does not have significant difference (P>0.05) compared with model control group, prescription 7 significantly can suppress thrombosis dry weight (P<0.05) compared with XINNAOXIN JIAONANG group, and except 1,2 prescription groups, all the other prescription groups do not have significant difference (P>0.05) compared with XINNAOXIN JIAONANG group.
4.2 XINNAOXIN JIAONANG prescriptions are on the impact of the Rabbit Blood Platelets maximum agglutination rate that ADP induces
By male rabbit, be divided into 12 groups at random, often organize 8.Be respectively the CMC-Na group of 0.5%, positive drug compound Salviae Miltiorrhizae group 148mg/kg, positive drug XINNAOXIN JIAONANG 100 mg/kg, prescription group 1 ~ 9 dosage be respectively 50,67,102,82,100,133,148,163,198mg/kg.Gavage every day (ig) is administered once, continuous gavage 7 days, administration volume is 5ml/kg, 0.5h carotid artery intubate blood-letting after last administration, the sodium citrate 1:9 anticoagulant of 3.8%, with the centrifugal 10min of 800r/min, get platelet rich plasma (PRP), remainder, with the centrifugal 10min of 3000r/min, gets platelet poor plasma (PPP), and aggregation inducing agent is with ADP (30 μ l) (final concentration 30 μm of ol/L).Often pipe 270 μ l PRP incubation 3min, then adds ADP induced aggregation, detects maximum platelet aggregation rate with LG-PABER-1 type platelet aggregation instrument.By following formulae discovery platelet aggregation inhibition rate:
The results are shown in Table 2.
From table 2, compound Salviae Miltiorrhizae group, XINNAOXIN JIAONANG group, other groups except prescription 1, all have certain inhibitory action to maximum platelet aggregation rate.Compared with model group, compound Salviae Miltiorrhizae group, XINNAOXIN JIAONANG group, prescription 3 all significantly can suppress anticoagulant (P<0.05), and prescription 2,4,5,6,7,8,9 groups can extremely remarkable anticoagulant (P<0.01); Prescription 1 does not have significant difference (P>0.05) compared with model group; Compared with XINNAOXIN JIAONANG group, compound Salviae Miltiorrhizae group, prescription 7 can remarkable anticoagulant (P<0.05).
. experimental analysis and conclusion
This experiment, by inducing rabbit platelet aggregation model analysis to rat arteriovenous shunt thrombosis and ADP, compares FUFANG DANSHEN PIAN, XINNAOXIN JIAONANG and composition of medicine prescription group and concentrates inhibiting height in thrombosis and platelet aggregation.In the middle of the model that rat arteriovenous shunt thrombosis is formed, XINNAOXIN JIAONANG group and composition of medicine prescription group show the effect of stronger inhibition thrombosis.In reduction wet weight of thrombus, the effect of prescription 7 is better than XINNAOXIN JIAONANG group, and prescription 4,5 groups and heart and brain glad group of Capsules group is similar; In reduction thrombosis dry weight, the effect of prescription 7 is better than XINNAOXIN JIAONANG group, and prescription 1,2,3,4,5,6 groups is not obvious to thrombosis inhibitory action; In rabbit platelet aggregation model, glad group of compound Salviae Miltiorrhizae group, heart and brain, other groups except prescription 1 group, all have stronger inhibitory action to maximum platelet aggregation rate, the effect of prescription 7 is better than compound Salviae Miltiorrhizae group and glad group of heart and brain, and prescription 1 acts on not obvious.Totally it seems, the inhibitory action of prescription 7 pairs of thrombosiss and platelet aggregation is better than other each positive drug group and composition of medicine prescription group.
for further clear and definite Radix Rhodiolae extract, Fructus Lycii extract, the effect of fresh syrup of Fructrs Hippophae powder extracts in prescription, we adopt L
9(3
4) result of orthogonal test to effect experiment analyze, with thrombosis dry weight suppression ratio (%), platelet aggregation inhibition rate for index, data statistics and interpretation of result as follows:
Observation index: thrombosis dry weight suppression ratio (%), platelet aggregation inhibition rate (%).
Direct-vision method is adopted to align friendship interpretation of result:
Factor A(Radix Rhodiolae extract)---factor A has pole appreciable impact to thrombosis dry weight suppression ratio, has a significant effect to platelet aggregation inhibition rate, and its level 3 is all better than other two levels, and the increase of Radix Rhodiolae extract amount is described, may improve drug effect.
Factor B(Fructus Lycii extract)---factor B does not have a significant effect to thrombosis dry weight suppression ratio, platelet aggregation inhibition rate.The amount of Fructus Lycii extract is described, may be little with drug effect dependency.
Factor C(fresh syrup of Fructrs Hippophae powder extracts)---factor C has some effects to thrombosis dry weight suppression ratio, has a significant effect to platelet aggregation inhibition rate, and its level 2,3 is all better than level 1, but no significant difference between level 2,3, illustrate that the amount of fresh syrup of Fructrs Hippophae powder extracts has impact to drug effect, but be increased to and to a certain degree will can not improve drug effect more afterwards.
Claims (7)
1. the composition of medicine of a Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, it is characterized in that, be made up of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, wherein Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts weight proportion are any a group in following weight proportioning: 644:85:9.66,644:170:2.41,644:340:4.83;
The composition of medicine preparation method of described a kind of Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts is: (1) Radix Rhodiolae extract technological process: after selecting clean, fragmentation, weigh, put in extraction pot, add 70% ethanol 10 times amount, 8 times amount respectively, reflux, extract, twice, 3 hours first times, second time 2 hours, merges twice reflux extracting liquid, after reclaiming ethanol, be concentrated into thick paste, dry, be ground into fine powder and namely obtain Radix Rhodiolae extract;
(2) Fructus Lycii extract technological process: through selecting only, weighing, putting in extraction pot, decoct with water twice, add water 10 times amount for the first time, and add water 8 times amount for the second time, and the time is respectively 2 hours, merge twice decocting liquid, filter, filtrate concentrates, with 80% ethanol precipitate with ethanol, centrifugal filtering, gets solid content drying, is ground into fine powder and namely obtains Fructus Lycii extract;
(3) fresh syrup of Fructrs Hippophae powder extracts technological process: fresh syrup of Fructrs Hippophae crosses concentrate drying after 80 mesh sieves, be ground into coarse powder, get coarse powder and add 70% ethanol 10 times amount, 8 times amount respectively, reflux, extract, twice, 3 hours first times, second time 2 hours, merge twice reflux extracting liquid, reclaim after ethanol, be concentrated into thick paste, dry, be ground into fine powder and namely obtain fresh syrup of Fructrs Hippophae powder extracts;
(4) said extracted thing is after assay, takes Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts by weight ratio respectively, make preparation after adding pharmaceutic adjuvant by preparation process according to content data;
Wherein: in Radix Rhodiolae extract, must not 1.2% be less than containing rhodioside, in Fructus Lycii extract, must not 3.5% be less than containing lycium barbarum polysaccharide with glucose meter, in fresh syrup of Fructrs Hippophae powder extracts, 20 .0% must not be less than containing total flavones in rutin.
2. the composition of medicine of a kind of Radix Rhodiolae extract according to claim 1, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, it is characterized in that, be respectively following weight scope containing the amount of rhodioside, lycium barbarum polysaccharide, total flavones in each dosage unit after Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts make composition of medicine: 4.32-8.64 mg, 3.25-12.99 mg, 0.483-1.931 mg.
3. the composition of medicine of a kind of Radix Rhodiolae extract according to claim 1, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, it is characterized in that, after Radix Rhodiolae extract, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts make composition of medicine in each dosage unit containing rhodioside, lycium barbarum polysaccharide, total flavones amount be respectively following weight combine in any one group: 4.32:6.49:1.931,8.64:3.25:1.931,8.64:6.49:0.483,8.64:12.99:0.965.
4. the composition of medicine of a kind of Radix Rhodiolae extract according to claim 1, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, is characterized in that, it is characterized in that its preparation is oral formulations.
5. the composition of medicine of a kind of Radix Rhodiolae extract according to claim 4, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, is characterized in that the pharmaceutic adjuvant containing oral formulations.
6. the composition of medicine of a kind of Radix Rhodiolae extract according to claim 4, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts, is characterized in that oral formulations comprises hard capsule, soft capsule, granule, tablet.
7. the composition of medicine of a kind of Radix Rhodiolae extract according to claim 1, Fructus Lycii extract and fresh syrup of Fructrs Hippophae powder extracts treats deficiency of both vital energy and Yin preparing, dizziness caused by stagnation of blood stasis, headache, cardiopalmus, asthma, weak, apply in the medicine of the erythrocytosis that anoxia causes and treatment, prevention myocardial infarction and cerebral infarction.
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