CN102908410A - Chinese rhubarb extract for treating osteoporosis and climacteric syndrome - Google Patents
Chinese rhubarb extract for treating osteoporosis and climacteric syndrome Download PDFInfo
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Abstract
The invention relates to a Chinese rhubarb extract for treating osteoporosis and climacteric syndrome. The Chinese rhubarb is rheum likiangense, wherein the content of stilbene total glycoside in the extract is more than or equal to 50 percent, and the stilbene total glycoside is rhaponticin, deoxyrhapontin or mixture of the rhaponticin and deoxyrhapontin.
Description
One. technical field
The invention belongs to field of medicaments, particularly a kind of Radix Et Rhizoma Rhei extract that is used for the treatment of osteoporosis and climacteric syndrome.
Two. technical background
Osteoporosis is a kind of systemic osteopathia, it is characterized in that unit volume bone amount descends, and the normal bone structure changes, and the osseous tissue microstructure is unusual, and showing as bone fragility increases, and the risk that causes fracturing increases, even slight wound is also fractured easily.Show as clinically lumbago and skelalgia, bow-backed deformity, and can fracture because of slight wound.Osteoporosis sickness rate in the crowd of old people and climacteric syndrome is higher, and especially postmenopausal women's sickness rate is higher, and this disease has become the especially frequently-occurring disease of elderly woman after the menopause of old people, and serious threat human healthy.
At present, the medicine for the treatment of osteoporosis mainly contains: 1. heavy dose of calcium preparation vitaminize D.2. hormone medicine.3. calcitonin.4. compound Chinese medicinal preparation.The therapy curative effect of heavy dose of calcium preparation vitaminize is poor, and complication is many; Life-time service estrogen can increase the risk that suffers from breast cancer with carcinoma of endometrium; Calcitonin needs injecting drug use, and is expensive, and side effect is many; Compound Chinese medicinal preparation treatment mechanism is not bery clear and definite, and curative effect is also unsatisfactory.Therefore, natural drug monomer or the effective site of exploitation treatment osteoporosis have important practical significance.
Climacteric syndrome is one of commonly encountered disease and frequently-occurring disease, mainly is the ovarian function decline, causes dyssecretosis, immunity degradation, autonomic nervous dysfunction causes a plurality of systems of whole body function and pathology pathological changes to occur, and the ovarian function decline is its core pathogenesis and pathologic basis.At present the climacteric syndrome prevalence is about about 90% in the middle aged and aged women, and wherein 50% needs Drug therapy, has become one of disease that affects middle aged and aged women quality of life.
Modern medicine study is thought, climacteric syndrome is because age growth, fertility, work hard, weak and sickly, the factors such as oophorectomize or radiotherapy, the excessive reduction of hormonal readiness causes the neurotransmitter due to hypothalamic-pituitary-ovarian axis or the dysadrenalism, hormone, cytokine etc. are unbalance to be the main cause of climacteric morbidity.So the relation of neuro-endocrine-immunoregulatory and climacteric syndrome is the closest.Doctor trained in Western medicine often adopts Hormone Replacement Therapy, although preferably curative effect is arranged, clinical practice finds that life-time service estrogen can increase the sickness rate of carcinoma of endometrium and breast carcinoma.Reproductive system Proliferation Lesion not only also can increase the risk of cardiovascular and cerebrovascular disease morbidity, thus significant limitation the clinical practice of this method.
Climacteric syndrome is commonly encountered diseases and frequently-occurring disease, also is a kind of disease for the treatment of and repeatedly showing effect than refractory, but also relates to the fields such as internal medicine, psychiatric department.At present, the drug main for the treatment of climacteric syndrome will be take estrogen and compound Chinese medicinal preparation as main, particularly life-time service estrogen can increase the risk that suffers from breast cancer with carcinoma of endometrium, and exploitation has bioactive natural drug monomer or effective site is significant for the treatment of climacteric syndrome.
Radix Et Rhizoma Rhei is one of Chinese medicine commonly used such as treatment constipation, inflammation and cancer.The main relieving constipation composition of genuine rhubarb is sennoside (dianthracene ketone) and anthraquinone glycoside.Compare with genuine rhubarb, non-genuine rhubarb anthraquinone content is lower and do not contain sennoside, so the relieving constipation effect is relatively poor.But non-genuine rhubarb diphenylethylene compounds content is higher, research is in recent years found, master in " non-certified products " Radix Et Rhizoma Rhei contains composition---ponticin and deoxidation ponticin, has multiple significant pharmacologically active, for example (Li JM, deoxidation ponticin suppress the glucose uptake of intestinal and kidney to blood sugar lowering: in vitro and in vivo research, pharmacological evaluation therapy (U.S.), 2007,320:38-46.); Blood fat reducing (Tadashi K, the natural grease polysaccharide inhibitor of from the Radix Et Rhizoma Rhei plant, finding, biological organic and pharmaceutical chemistry (U.S.), 2001,9:1887-1893.); Antitumor (Lee SH, induce human leukemia HL-60 cell's apoptosis, plant amedica (Germany), 2002.68:123-127 by the deoxidation ponticin.) etc.
Radix et Rhizoma Rhei lijiangensis is non-genuine rhubarb, is a kind of perennial medium-sized herbaceous plant.Be born under the height above sea level 2500-4000 rice alpine forest or shrub meadow.Be distributed in the ground such as Sichuan, Yunnan and Tibet.Contain emodin (emodin) in the Radix et Rhizoma Rhei lijiangensis root and rhizome, aloe-emodin (aloe-enodin), physcione (physcion), anthraquinone component and the tannins such as chrysophanol (chrysopharol).But the root hyoscine of plant has promoting blood circulation and hemostasis, eliminating stasis to stop pain, the effects such as dysentery relieving.
Chinese patent: 201110404970.4 disclose a kind of hepatoprotective effect of Radix et Rhizoma Rhei lijiangensis extract, and its main component is diphenyl ethene compounds.Other has bibliographical information, and stilbene glucoside has ability and the anti-allergic effects of removing ultra-oxygen anion free radical (O), but stilbene total glycosides treatment osteoporosis and climacteric syndrome yet there are no report both at home and abroad.The present invention has found the new purposes of this kind of Radix et Rhizoma Rhei lijiangensis extract through research.
Radix Et Rhizoma Rhei has had the use of more than one thousand years historical as a kind of medicinal herbs most in use in China, aboundresources, safety is without any problem, extract the treatment that the effective site stilbene total glycosides is used for osteoporosis and climacteric syndrome from Radix et Rhizoma Rhei lijiangensis, it is significant that it is developed as this sick medicine for the treatment of.The stilbene glycosides compound is different from the medicine that is used for the treatment of at present osteoporosis and climacteric syndrome, is a kind of this sick natural organic-compound that is used for the treatment of of recruit's structure type, and is significant to the further investigation of this compounds pharmacologically active.
Three. summary of the invention
Main purpose of the present invention provides the application of a kind of Radix Et Rhizoma Rhei extract in treatment osteoporosis and climacteric syndrome.
Radix Et Rhizoma Rhei extract of the present invention, the weight content of stilbene total glycosides 〉=50% in the extract.Stilbene total glycosides content 〉=70% in the preferred extract.Stilbene total glycosides content 〉=90% in the preferred extract.Stilbene total glycosides content 〉=95% in the most preferred extract.
In the stilbene total glycosides of the present invention, contain ponticin, deoxidation ponticin, perhaps both mixture.The mixture of ponticin and deoxidation ponticin wherein, both part by weight are: 1:10 to 10:1, the ratio of preferred ponticin and deoxidation ponticin is 1-3:1, particularly preferred is 1.5-2.5:1.
Radix Et Rhizoma Rhei extract of the present invention, wherein said Radix Et Rhizoma Rhei are any Radix Et Rhizoma Rhei class plant or its analog that contains above-mentioned stilbene total glycosides composition, such as Radix Polygoni Multiflori etc.The Radix Et Rhizoma Rhei class plant that preferably the stilbene total glycosides component content is higher, preferably Radix et Rhizoma Rhei lijiangensis, so Radix Et Rhizoma Rhei extract of the present invention is the Radix et Rhizoma Rhei lijiangensis extract most preferably.
Radix Et Rhizoma Rhei extract of the present invention, wherein said extract is standby in order to the below legal system:
Get the rhizome of Radix et Rhizoma Rhei lijiangensis, use ethanol extraction, concentrated through Recycled ethanol, to filter, drying and other steps obtains the crude product Radix Et Rhizoma Rhei extract, and through using dissolve with ethanol, activated carbon decolorizing filters again, and concentrated, the steps such as crystallization obtain elaboration Radix Et Rhizoma Rhei extract of the present invention,
The preparation method of preferred Radix Et Rhizoma Rhei extract of the present invention is as follows:
Radix Et Rhizoma Rhei is pulverized, 50-90% ethanol (v/v) with 5-10 times of weight extracts 1-3 time, and extracting solution is concentrated into without ethanol, crystallization, filtration drying, get the Radix Et Rhizoma Rhei crude extract, with 70-80% ethanol (v/v) dissolving of Radix Et Rhizoma Rhei crude extract with the 10-30 times of weight, add the active carbon reflux decolour, filter to get destaining solution, destaining solution is concentrated into without ethanol, and the room temperature spontaneous nucleation namely gets Radix Et Rhizoma Rhei extract of the present invention after the drying.
The preparation method of particularly preferred Radix Et Rhizoma Rhei extract of the present invention is as follows:
After Radix Et Rhizoma Rhei powder is broken to 40 orders, with 8 times of 70% ethanol room temperature supersound extraction 1 hour, extract again once by above method, extracted twice liquid is merged rear in being evaporated to without ethanol below 50 ℃, room temperature is placed crystallization in 24 hours, filtering for crystallizing and in vacuum drying below 45 ℃, get the Radix Et Rhizoma Rhei crude extract, the Radix Et Rhizoma Rhei crude extract is dissolved with 20 times of 75% alcohol reflux, added the active carbon reflux decolour 30 minutes, and filtered to get destaining solution, destaining solution is in being evaporated to without ethanol below 50 ℃, room temperature spontaneous nucleation 24 hours, crystallization namely gets Radix Et Rhizoma Rhei extract in vacuum drying below 45 ℃.
The present invention also comprises provides a kind of pharmaceutical composition that contains Radix Et Rhizoma Rhei extract of the present invention.Described pharmaceutical composition is any pharmaceutically useful dosage form, comprise oral, part, the dosage form of the administering modes such as injection, the preferred oral dosage form, as be selected from: the dosage form of tablet, capsule, suppository, granule, powder, patch, unguentum, pill or drop pill.Pharmaceutical composition of the present invention can according to the routine techniques preparation of pharmaceutics, as Radix Et Rhizoma Rhei extract of the present invention is mixed with the medicine acceptable carrier, be prepared into corresponding preparation.
Main purpose of the present invention is, a kind of new medical application is provided, i.e. the application of Radix Et Rhizoma Rhei extract of the present invention in the medicine of preparation treatment and prevention of osteoporosis disease.And, the application of Radix Et Rhizoma Rhei extract of the present invention in the medicine of preparation treatment and prevention climacteric syndrome.The application of Radix Et Rhizoma Rhei extract of the present invention in the medicine of preparation promotion Oesteoblast growth.Preferably treat and prevent postmenopausal osteoporosis.
Radix Et Rhizoma Rhei extract of the present invention, as active constituents of medicine be wherein stilbene total glycosides compound, mainly comprise two kinds of compositions in this compounds: ponticin and deoxidation ponticin, wherein the molecular structural formula of ponticin is as follows:
The molecular structural formula of deoxidation ponticin is as follows:
Carbon spectrum and hydrogen spectrum to the deoxidation ponticin are measured nuclear magnetic resonance data such as following table:
The nuclear magnetic resonance data of deoxidation ponticin
For the content of stilbene total glycosides in the labor extract of the present invention and wherein ponticin and deoxidation ponticin content, the invention also discloses a kind of content assaying method, said method comprising the steps of:
(1) required instrument and equipment
Agilent1260 high performance liquid chromatograph (being furnished with: quaternary pump, standard type automatic sampler, column oven, Agilent DAD detector); Sai Duolisi TE313S-DS type electronic balance; Excellent general UPT-2-40L type pure water preparation machine; KQ-500B type ultrasonic degas machine.
(2) required reagent
The Rhapontin(ponticin) (content 〉=98% is available from the Wei Keqi of Sichuan Province bio tech ltd, lot number: 120420) for standard substance; Desoxyrhaponticin(deoxidation ponticin) standard substance (content 〉=98%, self-control, lot number: 110804); Ethanol is analytical pure, Tianjin chemical industry all generations company limited; Methanol, acetonitrile are chromatographically pure, chemical industry branch company of Shandong YuWang Industry Co.,Ltd; Water is ultra-pure water, self-control.
(3) solution preparation
Standard solution: precision takes by weighing Rhapontin (ponticin) and Desoxyrhaponticin(deoxidation ponticin respectively) reference substance 3.0 mg and 5.3 mg, place 10 mL volumetric flasks with 50% dissolve with ethanol and be settled to scale, shake up, namely get reference substance solution.
The extract sample solution: the sample that precision takes by weighing certain mass places volumetric flask, with 50% dissolve with ethanol standardize solution, is mixed with certain density sample solution, makes it concentration within the range of linearity.
(4) determination step
1. high-efficient liquid phase chromatogram condition
Chromatographic column: Agilent ZORBAX Eclipse Plus C18(4.6mm * 100mm, 3.5 μ m); Column temperature: 25 ℃; Mobile phase: methanol-acetonitrile-water gradient elution, elution program sees the following form; Flow velocity: 1.0 mL/min; Detect wavelength: 320 nm.
Table 1 elution program
Table1?Elution?program
2. the preparation of standard curve
With the high performance liquid chromatograph automatic sampler respectively accurate sample introduction 2,4,6,8,10 μ L standard solutions analyze mensuration, with peak area Y sample size X (μ g) is carried out linear regression, get regression equation.Rhapontin (ponticin): Y=3910.7X-25.85(R
2=1); Desoxyrhaponticin(deoxidation ponticin): Y=4204.8X+50.137(R
2=0.9997).
3. test liquid assay
With the filtering with microporous membrane of sample solution with 0.45 μ m, 10 μ L analyze mensuration according to the high-efficient liquid phase chromatogram condition sample introduction, with external standard method Rhapontin(ponticin in the calculation sample solution respectively) and Desoxyrhaponticin(deoxidation ponticin) content.
4. computing formula
Rhapontin(ponticin in the sample) and Desoxyrhaponticin(deoxidation ponticin) content (g/100g) computing formula be:
C（g/100g）=（Cx×V）×100/Ms
In the formula: the Rhapontin(ponticin in Cx-sample solution) and Desoxyrhaponticin(deoxidation ponticin) concentration (mg/mL);
V-sample solution volume, mL;
Ms-take by weighing sample quality (mg).
Below data further specify beneficial effect of the present invention by experiment.
For the medical application to Radix Et Rhizoma Rhei extract of the present invention provides support, the present invention has carried out following Research experiment, and Radix Et Rhizoma Rhei extract of the present invention is used for external osteoblast increment experiment.Radix Et Rhizoma Rhei extract of the present invention is used for the experiment of castration mouse autonomic activities.Radix Et Rhizoma Rhei extract of the present invention is used for Ovariectomized Rat Serum estradiol impact experiment.Below be experiment content:
The external osteoblastic proliferation experiment of Radix Et Rhizoma Rhei extract of the present invention is as follows:
1, experiment purpose
Take estrogen receptor as action target spot, take the strain of MC3T3-E1 osteoblast as the active model of investigating, estrogen with synthetic---diethylstilbestrol is as positive control drug, take osteoblastic propagation level as index, the study tour Radix Et Rhizoma Rhei extract is for the effect that promotes osteoblastic proliferation and the ability of formation, and clear and definite its dose-effect relationship, thereby provide foundation for preventing and treating osteoporotic natural drug research and development.
2, the preparation of tested material
2.1 positive control drug:
Diethylstilbestrol is purchased from Shanghai and grinds the territory bio tech ltd.Storing solution concentration 1 * 10
-2Mol/L, cell sample solution final concentration 1 * 10
-7Mol/L.
2.4 Radix Et Rhizoma Rhei extract: (preparation of the embodiment of the invention 1 method)
Made by Qinghai Yi Naweikang bio tech ltd, stilbene total glycosides content is 95.68%, and wherein, ponticin content is 62.77%, and deoxidation ponticin content is 32.91%.
Storing solution concentration 1mg/mL, cell sample solution final concentration are respectively 100 μ g/mL, 10 μ g/mL, 1 μ g/mL.
3, experimental technique
The MC3T3-E1 cell is purchased from Shanghai and grinds brilliant industry Science and Technology Ltd..Get the MC3T3-E1 cell and be inoculated in 96 well culture plates, behind the constant temperature culture 24h, the blank group is established in administration simultaneously, continues to cultivate 48h, cultivates and finishes the front MTT of adding, measures the OD value in every hole after the dyeing in 490nm.
4, experimental result and discussion
Annotate: compare * p＜0.05 with the blank group, * * p＜0.01
The result shows:
The Radix Et Rhizoma Rhei extract group of 100 μ g/mL, 10 μ g/mL, 1 μ g/mL all has proliferation function to osteoblast, and wherein, concentration is that the Radix Et Rhizoma Rhei extract group of 100 μ g/mL (P＜0.05) has remarkable proliferation function to osteoblast.
Other experiments of Radix Et Rhizoma Rhei extract of the present invention are as follows:
1. on the impact of castration mouse autonomic activities
1.1 medicine: Radix Et Rhizoma Rhei extract (preparation of the embodiment of the invention 1 method), made by Qinghai Yi Naweikang bio tech ltd; Nilestriol Tablets is made by the Beijing Sihuan Pharmaceutical Co., Ltd.
1.2 animal: the Kunming small mouse, body weight 18-22 gram, female, provided by Guangdong Medical Lab Animal Center.
1.3 ZZ-6 mice autonomic activities analyzer, Chengdu TME Technology Co., Ltd. produces.
1.4 method
Method of emasculation: get healthy mice, behind etherization, extract after the bilateral ovaries and to carry out one by one vaginal smear examination on the 5th day, once a day, continuous five days, get ovariectomy and be used for experiment without variation person's (castration group) emotionally fully.False castration treated animal operation is except not extracing the ovary, and is remaining with the castration group.
Female mice is got 12 castration groups that rehearse, and the animal of passed examination is used for experiment after the remaining castration.Castration mouse is divided into five groups at random, 12 every group.False castration group, castration group all give normal saline, the positive controls gavage gives nilestriol 20 microgram per kilograms, high, medium and low dosage group respectively gavage gives 40 milligrams of per kilograms of Radix Et Rhizoma Rhei extract, 20 milligrams of per kilograms, 10 milligrams of per kilograms, in postoperative beginning in the 9th day administration, once a day, continuous 30 days, after the last administration one hour, mice is put into ZZ-6 mice autonomic activities analyzer, measure the autonomic activities number of times in ten minutes.Statistical analysis is carried out in the data t check between group.
1.5 experimental result is as follows
Table 1 Radix Et Rhizoma Rhei extract on the impact of castration mouse autonomic activities (n=12,
)
Compare * p＜0.05, * * p＜0.01 with the castration group
Experimental result shows, compares with the castration group, and the autonomic activities number of times of false castration group mice obviously reduces, and has very significant meaning (* * p＜0.01), proves the modeling success.Compare with the castration group, the autonomic activities number of times of positive controls, Radix Et Rhizoma Rhei extract high dose group mice obviously reduces, and has very significant meaning (* * p＜0.01); The autonomic activities number of times of amount group mice obviously reduces in the Radix Et Rhizoma Rhei extract, and has significant (* p＜0.05).The above results shows that Radix Et Rhizoma Rhei extract can significantly reduce the castration mouse autonomic activities, illustrates that it has good sedation.
2. on the impact of Ovariectomized Rat Serum estradiol
2.1 medicine: extract (preparation of the embodiment of the invention 1 method), produced by Qinghai Yi Naweikang bio tech ltd; Nilestriol Tablets is made by the Beijing Sihuan Pharmaceutical Co., Ltd; Human serum estradiol (E2) radioimmunoassay kits, Tianjin Xin Wan bio tech ltd is produced.
2.2 animal: female Wistar rats, at 12 monthly ages, body weight is the 260-320 gram, is provided by Guangdong Medical Lab Animal Center.
2.3 UV759 type ultraviolet-visible spectrophotometer, Shanghai Precision Scientific Apparatus Co., Ltd makes.
2.4 method
Method of emasculation: get healthy rat, behind the etherization, extract after the bilateral ovaries and to carry out one by one vaginal smear examination on the 5th day, once a day, continuous five days, get ovariectomy and be used for experiment without variation person's (castration group) emotionally fully.False castration treated animal operation is except not extracing the ovary, and is remaining with the castration group.
Female rats is got 12 castration groups that rehearse, and the passed examination animal is used for experiment after the remaining castration.Castrated rats is divided into five groups at random, 12 every group.False castration group, castration group, all give normal saline, the positive controls gavage gives nilestriol 10 microgram per kilograms, and high, medium and low dosage group respectively gavage gives Radix Et Rhizoma Rhei extract 40 milligrams of per kilograms, 20 milligrams of per kilograms, 10 milligrams of per kilograms are in postoperative beginning in the 9th day administration, once a day, continuous 30 days, one hour excision eyeball is got blood after the last administration, measures serum estradiol concentration by the requirement of test kit description, and experimental data adopts the t check to carry out statistical analysis between group.
2.5 experimental result is as follows
Radix Et Rhizoma Rhei extract to Ovariectomized Rat Serum (E2) concentration as influencing factor (n=12,
)
Compare * p＜0.05, * * p＜0.01 with the castration group
The results show is compared with the castration group, and false castration group rat blood serum estradiol (E2) concentration obviously raises, and has very significant meaning (* * p＜0.01), shows the modeling success.Compare with the castration group, positive controls, Radix Et Rhizoma Rhei extract high dose group serum estradiol (E2) concentration obviously raise, and has very significant meaning (* * p＜0.01), dosage group Ovariectomized Rat Serum estradiol (E2) concentration that can obviously raise in the Radix Et Rhizoma Rhei extract, and have significant (* p＜0.05).Above-mentioned experimental result shows, Radix Et Rhizoma Rhei extract Ovariectomized Rat Serum estradiol (E2) concentration that can significantly raise.
3 in sum, and Radix Et Rhizoma Rhei extract can significantly improve the estrogen level of castrated rats, has simultaneously good sedation, and these effects are pharmacological basis for the treatment of climacteric syndrome.
Four, the specific embodiment:
Further specify by the following examples the present invention, but not as limitation of the present invention.
Embodiment 1
The preparation method of Radix Et Rhizoma Rhei extract is as follows:
50 kilograms of Radix et Rhizoma Rhei lijiangensis are ground into 40 orders, add 400 kilogram of 70% ethanol and use multifunction ultrasonic extraction pot supersound extraction 1 hour, extract again once by above method, merge extracted twice liquid, alcohol-free in being evaporated to below 50 ℃, the concentrated solution room temperature is placed 24 hours spontaneous nucleations, filtering for crystallizing and in ℃ below vacuum drying, get 4.6 kilograms of Radix Et Rhizoma Rhei crude extracts, this crude extract is added 92 kilogram of 75% reflow of alcohol dissolving, added 4.6 kilogram of 767 type active carbon reflux decolour 30 minutes, filtered while hot, filtrate is alcohol-free in being evaporated to below 50 ℃, concentrated solution room temperature spontaneous nucleation 24 hours, filter to get crystallization, this crystallization in vacuum drying below 45 ℃ 8 hours, is got 4.1 kilograms of Radix Et Rhizoma Rhei extracts.
Embodiment 2
Radix Et Rhizoma Rhei extract to embodiment 1 detects:
One, Radix Et Rhizoma Rhei extract of the present invention has following character:
(1) character: this product is that off-white color is to khaki crystallization or amorphous powder
(2) dissolubility: be slightly soluble in water, be dissolved in methanol, ethanol, be insoluble to benzene, ether, chloroform
(3)pH：?6.5-7.5
(4) ultra-violet (UV) band absorbs: λ max=325nm
(5) color reaction:
Aminoacid (ninhydrin reaction): feminine gender
Protein (trichloroacetic acid method): feminine gender
Phenol (ferric chloride test method): the positive
Two, the assay of effective ingredient in the Radix Et Rhizoma Rhei extract of the present invention:
Adopt detection method provided by the invention, the result of detection is as follows:
Stilbene total glycosides content: 95.68%
Ponticin content: 62.77%
Deoxidation ponticin content: 32.91%
Embodiment 3
Radix et Rhizoma Rhei lijiangensis is ground into granule, 50% ethanol (v/v) diafiltration with 5 times of weight is extracted, extracting solution is evaporated to without ethanol and distinguishes the flavor of, cool to room temperature, place 12 hours crystallizations, filtration drying, the Radix Et Rhizoma Rhei crude extract, the Radix Et Rhizoma Rhei crude extract is dissolved with 70% ethanol (v/v) of 10 times of weight, add the active carbon reflux decolour 1 hour of 1% weight, filter to get destaining solution, destaining solution is concentrated into without ethanol flavor, room temperature natural cooling 12 hours, crystallization, filter, namely get Radix Et Rhizoma Rhei extract of the present invention after the drying.
Embodiment 4
Radix et Rhizoma Rhei lijiangensis is ground into granule, with 90% ethanol (v/v) reflux, extract, of 10 times of weight 3 times, the each backflow 1 hour, three times extracting solution merges, and is evaporated to without the ethanol flavor, is cooled to 4 ℃, place 24 hours crystallizations, filtration drying gets the Radix Et Rhizoma Rhei crude extract, with 80% ethanol (v/v) dissolving of Radix Et Rhizoma Rhei crude extract with 30 times of weight, the active carbon reflux decolour 0.5 hour that adds 0.5% weight, filter to get destaining solution, destaining solution is concentrated into without the ethanol flavor, is cooled to 4 ℃, cooled off 24 hours, crystallization filters, and namely gets Radix Et Rhizoma Rhei extract of the present invention after the drying.
Embodiment 5
Radix Et Rhizoma Rhei powder is broken into fine powder, with 80% ethanol (v/v) reflux, extract, of 6 times of weight 3 times, merge extractive liquid,, be concentrated into without the ethanol flavor, be cooled to 10 ℃, placed 48 hours, crystallization, filtration drying gets the Radix Et Rhizoma Rhei crude extract, the Radix Et Rhizoma Rhei crude extract with the dissolving of 75% ethanol (v/v) of 15 times of weight, is added the active carbon reflux decolour 2 hours of 1% weight, filter to get destaining solution, destaining solution is concentrated into without ethanol and distinguishes the flavor of, cool to room temperature, spontaneous nucleation namely gets Radix Et Rhizoma Rhei extract of the present invention after the drying.
Claims (10)
1. a Radix Et Rhizoma Rhei extract is characterized in that, stilbene total glycosides content 〉=50% in this extract.
2. according to claim 1 Radix Et Rhizoma Rhei extract is characterized in that described stilbene total glycosides is ponticin, deoxidation ponticin or both mixture, wherein stilbene total glycosides content 〉=70%.
3. according to claim 1 Radix Et Rhizoma Rhei extract is characterized in that in the mixture of described ponticin and deoxidation ponticin, both ratios are: 1:10 to 10:1, wherein stilbene total glycosides content 〉=90%.
4. according to claim 1 Radix Et Rhizoma Rhei extract is characterized in that wherein said Radix Et Rhizoma Rhei is Radix et Rhizoma Rhei lijiangensis, wherein stilbene total glycosides content 〉=95%.
5. according to claim 1 Radix Et Rhizoma Rhei extract is characterized in that wherein said extract is standby in order to the below legal system:
Radix Et Rhizoma Rhei is pulverized, with the 50-90% ethanol extraction of 5-10 times of weight 1-3 time, extracting solution is concentrated into without ethanol, crystallization, filtration drying, get the Radix Et Rhizoma Rhei crude extract, with the 70-80% dissolve with ethanol of Radix Et Rhizoma Rhei crude extract with the 10-30 times of weight, add the active carbon reflux decolour, filter to get destaining solution, destaining solution is concentrated into without ethanol, and the room temperature spontaneous nucleation namely gets Radix Et Rhizoma Rhei extract of the present invention after the drying.
6. according to claim 1 Radix Et Rhizoma Rhei extract is characterized in that wherein said extract is standby in order to the below legal system:
After Radix Et Rhizoma Rhei powder is broken to 40 orders, with 8 times of 70% ethanol room temperature supersound extraction 1 hour, extract again once by above method, extracted twice liquid is merged rear in being evaporated to without ethanol below 50 ℃, room temperature is placed crystallization in 24 hours, filtering for crystallizing and in vacuum drying below 45 ℃, get the Radix Et Rhizoma Rhei crude extract, the Radix Et Rhizoma Rhei crude extract is dissolved with 20 times of 75% alcohol reflux, added the active carbon reflux decolour 30 minutes, and filtered to get destaining solution, destaining solution is in being evaporated to without ethanol below 50 ℃, room temperature spontaneous nucleation 24 hours, crystallization namely gets Radix Et Rhizoma Rhei extract in vacuum drying below 45 ℃.
7. pharmaceutical composition that contains the described Radix Et Rhizoma Rhei extract of claim 1 of effective dose.
8. according to claim 1 pharmaceutical composition is characterized in that described pharmaceutical composition is selected from: tablet, capsule, suppository, granule, powder, patch, unguentum, pill or drop pill.
9. the application of the Radix Et Rhizoma Rhei extract of claim 1 in the medicine of preparation treatment osteoporosis and promotion Oesteoblast growth.
10. the application of the Radix Et Rhizoma Rhei extract of claim 1 in the medicine of preparation treatment climacteric syndrome.
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