CN1981811B - Danshen effective component, its preparation, making method and usage - Google Patents
Danshen effective component, its preparation, making method and usage Download PDFInfo
- Publication number
- CN1981811B CN1981811B CN2005101223489A CN200510122348A CN1981811B CN 1981811 B CN1981811 B CN 1981811B CN 2005101223489 A CN2005101223489 A CN 2005101223489A CN 200510122348 A CN200510122348 A CN 200510122348A CN 1981811 B CN1981811 B CN 1981811B
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- solution
- ethyl acetate
- sample
- acetonitrile
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
An active component of red sage root is just dihydrotanshinone I. Its preparing process includes such steps as breaking red sage root, adding ethyl acetate and alcohol, thermal extracting, concentrating to obtain extract, mixing it with silicon gel, separating with positive-phase silicon gel column, washing with petroether and ethyl acetate, eluting with chloroform and methanol, concentrating theeluting liquid, drying, and separating by liquid-phase chromatography. Its application is also disclosed.
Description
Technical field
The present invention relates to a kind of Chinese medicine extract for the treatment of cardiovascular disease, relate in particular to the active component that from Radix Salviae Miltiorrhizae, extracts, preparation and preparation method thereof and purposes, this active component mainly contains dihydrotanshinone I (Dihydrotanshinone I).
Background technology
Coronary heart disease and angina pectoris are harm humans health " first killers ", in recent years, along with the variation of China's population senescence and people's work, life, dietary structure and environment etc., the incidence rate of cardiovascular and cerebrovascular diseases such as coronary heart disease also increases year by year, and the people's physical and mental health in serious threat.In the natural product many active substances have resist myocardial ischemia, anoxia functions, some of them have been developed to treatment coronary heart disease and anginal new drug, as treat the medicine DIAOXINXUE KANG of coronary heart disease, it is by 8 kinds of pure Chinese medicinal preparations that steroid saponin is made that extract from Chinese medicine; Flavonihippophae is to be that raw material extracts the pure natural medical that processes with the Fructus Hippophae, and its effective ingredient is a Fructus Hippophae total flavones.Thereby from natural product, seek have resist myocardial ischemia, the active substance of anoxia physiologically active, be find, one of effective way of developing new drug.China's medicinal organism resource is very abundant, and its biological active substances is research and finds new drug guide chemicals, the natural treasure-house of developing new drug.At present, China extracts active substance from natural product, be used to be developed to treatment coronary heart disease, safety is good, toxicity is low new drug also seldom, from natural product, extract active substance, be developed to have and resist myocardial ischemia, be used for the treatment of coronary heart disease and anginal new drug, have significant application value and wide development prospect.
Radix Salviae Miltiorrhizae, another name blood is taken root, red, Herba Wedeliae Wallichii, Arisaema balansae Engl..Radix Salviae Miltiorrhizae is from plant Salvia miltiorrhiza rhizome.Radix Salviae Miltiorrhizae is used as medicine and begins to be stated from the Shennong's Herbal of Han dynasty.The traditional Chinese medical science is thought, its bitter in the mouth cold nature has promoting blood circulation to restore menstrual flow, an effect of the silt pain relieving of dispelling, the relieving restlessness that clears away heart-fire, removing heat from blood eliminating carbuncle, is applicable to the treatment menoxenia, coronary heart disease, arteriosclerosis, hyperlipidemia, angina pectoris, dysmenorrhea, amenorrhea, metrorrhagia, leukorrhagia, traumatic injury blood stasis, insomnia, hepatitis B, neurasthenia, the schistosomicide hepatosplenomegaly, diseases such as arthralgia, particularly gynecological, internal medicine and external wounds card belongs to the blood stasis hot person that holds concurrently and uses always.Modern study shows, mainly contain Tanshinone I, IIA, IIB (Tanshinone I in the Radix Salviae Miltiorrhizae, IIA, IIB), iso tanshinone I, IIA (Isotanshinone I, IIA), cryptotanshinone (Cryptotanshinone), different cryptotanshinone (Isocryptotanshinone), methyltanshinone (Methyltanshinone), hydroxyl TANSHINONES etc.
Radix Salviae Miltiorrhizae is the common drug of treatment cardiovascular disease, and in recent decades, research institution has carried out a large amount of research to Radix Salviae Miltiorrhizae both at home and abroad, and the various patent medicine that contain red rooted salvia are developed diseases such as being used for the treatment of coronary heart disease, angina pectoris, cerebral infarction.Radix Salviae Miltiorrhizae has the influence of cardiovascular system: (1) increases coronary flow.But composition such as Radix Salviae Miltiorrhizae, element coronary blood flow increasing, promote collateral circulation, improve myocardium microcirculation, do not increase ventricular stroke work and myocardial oxygen consumption simultaneously.(2) microcirculation improvement.Radix Salviae Miltiorrhizae has the effective ingredient of microcirculation improvement.In addition, effective ingredient can influence multiple thrombin, improves hemorheological property, can reduce plasma viscosity, regulate cell electrophoresis rate and packed cell volume patients with coronary heart disease in the Radix Salviae Miltiorrhizae, microcirculation improvement has therapeutical effect preferably to blood stasis patient's blood " sticking, poly-, stagnate " tendency.Studies show that Radix Salviae Miltiorrhizae can anticoagulant and antithrombotic effect.Just because of Radix Salviae Miltiorrhizae has above pharmacological action to cardiovascular system, therefore, from Radix Salviae Miltiorrhizae, isolate active component, the modern Chinese medicine of developing the treatment cardiovascular disease has great importance.
Summary of the invention
The object of the present invention is to provide a kind of Danshen effective component.
Another object of the present invention is to provide the preparation method of this Danshen effective component.
The present invention also provides the preparation that contains this Danshen effective component and the purposes of this component.
The invention provides following technical scheme:
Danshen effective component of the present invention mainly contains dihydrotanshinone I.Wherein, by weight percentage, the content of dihydrotanshinone I is 96.5～98%.
Preferably, in the Danshen effective component of the present invention, the content of dihydrotanshinone I is 96.8～97.7%.
Best, in the Danshen effective component of the present invention, the content of dihydrotanshinone I is 97.0～97.5%.
The preparation method of Danshen effective component of the present invention, comprise the following steps: that red rooted salvia is pulverized the back adds ethyl acetate and ethanol, heating extraction gets extracting solution, and extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate, get eluent I as mobile phase, abandon it, change chloroform and methanol then as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a semi-preparative column, and mobile phase is water and acetonitrile, gradient elution.
Preferably, Danshen effective component preparation method of the present invention, comprise the following steps: to get red rooted salvia, it is pulverized the back add ethyl acetate and ethanol (1: 0.8～1.2), reflux 0.8～1.2 hour, extract 1～3 time, merging filtrate gets extracting solution, and extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate (47～53:, get eluent I 1) as mobile phase, abandon it, change chloroform and methanol (8～13: 1) as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying then; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a semi-preparative column, and mobile phase is water and acetonitrile, and gradient elution, flow velocity are 2.8～3.2ml/min, and column temperature is a room temperature.
Best, Danshen effective component preparation method of the present invention, comprise the following steps: to get red rooted salvia, it is pulverized the back add ethyl acetate and ethanol (1: 1), reflux 1 hour, extract 2 times, merging filtrate gets extracting solution, and extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate (50: 1), get eluent I as mobile phase, abandon it, change chloroform and methanol (10: 1) then as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is semi-preparative column (Lichrospher C18; 10.0mm * 250mm, 5 μ m), mobile phase is water (A) and acetonitrile (B), and the gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 50% aqueous solution, Mobile phase B are 50% acetonitrile solution; In the time of 35 minutes, mobile phase A is that 20% aqueous solution, Mobile phase B are 80% acetonitrile solution; In the time of 45 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; In the time of 55 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; Flow velocity is 3ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 21.7～30.3min, and solution obtains active component behind concentrate drying.
Danshen effective component of the present invention can be used as active component, adds the adjuvant of accepting on the pharmaceutics, makes preparation according to the preparation method of the preparation of putting down in writing on the pharmaceutics.
Danshen effective component of the present invention can also add the adjuvant of accepting on the pharmaceutics as one of active component, makes preparation according to the preparation method of the preparation of putting down in writing on the pharmaceutics.
Described preparation comprises injection, drip liquid, injectable powder, granule, tablet, electuary, powder, oral liquid, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, sucks agent, granule, pill, unguentum, sublimed preparation, spray, drop pill, disintegrating agent, oral cavity disintegration tablet, micropill etc.
Danshen effective component of the present invention and preparation thereof can be used in preparation treatment, angiocardiopathy preventing medicine.
Beneficial effect of the present invention is:
1. use normal phase silicagel column in the extraction and separation process of the present invention, can remove impurity such as desaccharide, protein, aminoacid effectively, improved content of effective, adopted preparative hplc simultaneously, can obtain effective ingredient fast and accurately.
2. Danshen effective component chemical constituent provided by the invention is simply clear and definite, is easier to illustrate its mechanism of action on pharmacological research, is easier to the quality control of medicine aborning.
Description of drawings
Fig. 1 is the HPLC analysis chart of Danshen effective component of the present invention.
The specific embodiment
Further describe flesh and blood of the present invention and beneficial effect below in conjunction with embodiment, this embodiment only is used to the present invention is described but not limitation of the present invention.
The preparation of embodiment one Danshen effective component
The 200g red rooted salvia is pulverized the back add ethyl acetate and ethanol, heating extraction gets extracting solution I, abandons it.Medicinal residues add ethanol, heating extraction gets extracting solution II, extracting solution II is condensed into extractum 12.74g, use dissolve with methanol extractum, adopt the ODS-C18 post that it is separated, at first use low concentration methanol as mobile phase, get eluent I, change high concentration methanol then as mobile phase, get eluent II, get sample 1.85g behind the concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a semi-preparative column, and mobile phase is water and acetonitrile, gradient elution, sample 100% dissolve with ethanol, separate through preparative liquid chromatography, collect solution, obtain active component 0.008g behind the concentrate drying at time period 21.7～30.3min.
The preparation of embodiment two Danshen effective components
The 500g red rooted salvia is pulverized the back add ethyl acetate and ethanol (1: 0.8～1.2), reflux 0.8～1.2 hour is extracted 1～3 time, and merging filtrate gets extracting solution I, abandons it.In medicinal residues, add 60%～80% ethanol, reflux 0.8～1.2 hour is extracted 1～3 time, filtrate merge extracting solution II, extracting solution II is condensed into extractum 31.82g, with 40%～60% dissolve with methanol extractum, adopt the ODS-C18 post that it is separated, at first use 3%～8% methanol as mobile phase, get eluent I, change 50%～70% methanol then as mobile phase, get eluent II, get sample 4.62g behind the concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a semi-preparative column, and mobile phase is water and acetonitrile, gradient elution, flow velocity is 2.8～3.2ml/min, column temperature is a room temperature, collects solution at time period 21.7～30.3min, and solution obtains active component 0.019g behind concentrate drying.
The preparation of embodiment three Danshen effective components
Get red rooted salvia 250g, it is pulverized the back add ethyl acetate and ethanol (1: 1), reflux 1 hour is extracted 2 times, filtrate merge extracting solution.Extracting solution is condensed into extractum 17.2g, with extractum and silica gel mixed sample, separate with normal phase silicagel column, at first use petroleum ether and ethyl acetate (50: 1) as mobile phase, get eluent I, change chloroform and methanol (10: 1) then as mobile phase, get eluent II, get sample 2.5g behind the concentrate drying.Continue to separate the sample that obtains with preparative liquid chromatography.The separation condition of preparative hplc: chromatographic column is semi-preparative column (the Lichrospher C18 of Chinese nation; 10.0mm * 250mm, 5 μ m), mobile phase is water (A) and acetonitrile (B), and the gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 50% aqueous solution, Mobile phase B are 50% acetonitrile solution; In the time of 35 minutes, mobile phase A is that 20% aqueous solution, Mobile phase B are 80% acetonitrile solution; In the time of 45 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; In the time of 55 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; Flow velocity is 3ml/min, and column temperature is a room temperature.Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 21.7～30.3min, and solution obtains Radix Salviae Miltiorrhizae extract 0.0104g behind concentrate drying
The analysis of embodiment four Danshen effective components
(1) assay of dihydrotanshinone I (high performance liquid chromatography) in the Radix Salviae Miltiorrhizae extract
Chromatographic conditionChromatographic column Agilent Zorbax SB-C18 post (4.6mm * 150mm, 5 μ m); Adopt gradient elution, mobile phase A is 0.2% glacial acetic acid aqueous solution mutually, and Mobile phase B is mutually for containing the acetonitrile solution of 0.2% glacial acetic acid; The gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 90% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 10% 0.2% glacial acetic acid; In the time of 10 minutes, mobile phase A is that 30% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 70% 0.2% glacial acetic acid; In the time of 30 minutes, mobile phase A is that 5% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 95% 0.2% glacial acetic acid; Flow velocity 0.5mL min-1; It is long to detect the wavelength all-wave; 30 ℃ of column temperatures; DAD detects in the interscan of 190-400nm scope; Sample size 5 μ L.
The preparation of need testing solutionTake by weighing this product, in volumetric flask, be diluted to scale, shake up, promptly with dissolve with methanol solution.
Assay methodThe accurate need testing solution of drawing injects chromatograph of liquid, measures, promptly.
(2) above-mentioned Radix Salviae Miltiorrhizae extract HPLC-ELSD finger printing
Assay methodReferring to the dihydrotanshinone I assay (high performance liquid chromatography) in (1) above-mentioned Radix Salviae Miltiorrhizae extract.The record chromatograph time is 25 minutes.
Analysis resultThe HPLC-ELSD analysis of spectra of Danshen effective component is seen Fig. 1.
The preparation of embodiment five drop pill
Get Danshen effective component 0.01g and 10.5g Polyethylene Glycol-6000 mix homogeneously of embodiment three, heating and melting moves in the drop pill drip irrigation behind the change material, and in ℃ liquid paraffin of medicine liquid droplet to 6～8, oil removing makes 200 of drop pill.
The preparation of embodiment six lyophilized injectable powders
Get Danshen effective component 0.01g, glucose 4.5g, sodium thiosulfate 0.9g and the distilled water 1ml of embodiment three, behind the said components mix homogeneously, lyophilization, 300 of packing, promptly.
The preparation of embodiment seven lyophilized injectable powders
Get Lignum Dalbergiae Odoriferae oil 1.5g, join in the saturated hydroxypropyl of 13ml, stirring and dissolving filters, and the filtrate cold drying gets the clathrate powder of Lignum Dalbergiae Odoriferae oil and hydroxypropyl.Except that above-mentioned Lignum Dalbergiae Odoriferae oil closes the clathrate powder of hydroxypropyl, get Radix Salviae Miltiorrhizae extract 0.01g, mannitol 5.5g, calcium disodium edetate 0.9g and the distilled water 2ml of embodiment three again, behind the said components mixing, lyophilization, 250 of packing, promptly.
The pharmacological evaluation of embodiment eight Danshen effective components
1 pharmacological model: neonatal rat myocardial cell hypoxic-ischemic model
Former generation myocardial cell cultivationAfter 1～3 day SD neonatal rat that is born was put to death, it was dirty to core, and subtracts into 1mm after PBS liquid cleans
3About fragment.With 0.125% trypsin Sigma, USA)+0.05% (Gibco is USA) in 37 degree digestion 3～4 times for collagenase II.The differential adherent method was separated into fibrocyte after 1.5 hours, and cell suspension is transferred to (every hole about 4 * 10 in 48 orifice plates
5Cell).(Gibco, (Falcon, USA) cell of cultivating 3-6 days supplies medicine efficacy screening USA) to add 10% hyclone through DMEM.Screen and replaced with serum-free medium in preceding 1 day.Cultivate 3～6 days myocardial cell, PBS washing back adds the pastille culture fluid.Place 95%N
2And 5%CO
2Anoxia is 6 hours in the incubator.Then at CO
2Reoxygenation is 3 hours in the incubator.Get cell conditioned medium liquid and measure LDH content.
Dosage regimenThe Danshen effective component that extraction obtains is made into 10 with sugar-free Hanks liquid
-4G/mL test liquid, liposoluble constituent can add a small amount of DMSO hydrotropy (the DMSO final concentration is controlled at below 0.2%).The final concentration of each component in culture fluid is controlled at 10
-5G/mL.
The lactic acid dehydrogenase assayLactic acid dehydrogenase content can be represented the cell injury degree in the cell culture fluid.The lactic acid dehydrogenase content that bleeds in the supernatant is high more, shows that cell injury is also remarkable.The determination of lactate dehydrogenase test kit builds up bio-engineering research institute available from Nanjing.Assay method is: get 30 microlitre cell conditioned medium liquid, add 50 microlitre substrate buffer and 10 microlitre lactic dehydrogenase enzyme cofactors, 37 degree vibrations 15 minutes add 50 microlitre 2,4 dinitrophenyl hydrazines again, 37 degree vibrations 15 minutes.Parallel blank.Extract reaction solution 50 microlitres and add 200 microlitre 0.4mol/L sodium hydroxide solutions, leave standstill 3～4 minutes after, measure shading value with microplate reader in 450nm.
Drug effect result
All data are represented with the mean value variance.Adopt monolateral variance analysis and T check carrying out statistical analysis.Compare with model group, P＜0.05 is thought significantly effectively.Drug effect the results are shown in Table 1.According to myocardial cell lactic acid dehydrogenase (LDH) testing result, Danshen effective component has the highly significant effect to reducing LDH release.
Table 1
2 pharmacological models: huve cell anoxia reoxygenation model
Former generation huve cell cultivationGet the healthy newborn umbilical cord, clean the interior residual bloodstain of vein blood vessel with 30～50mlHanks liquid.Look the 0.1% collagenase I that umbilical cord length adds respective amount, change incubator digestion 15～20 minutes over to.Subsequently Digestive system in the umbilical cord is carefully collected in the beaker, and with Hanks liquid with beaker rinse twice, collect in the lump and be sub-packed in centrifuge tube in the beaker, centrifugal 10 minutes of 900rpm.Supernatant is removed in suction, with the abundant dissolution precipitation of M199 culture fluid (contain 10% hyclone, 100U/ml is two anti-, 0.15mg/ml Heparin, 10uM VC).Gained cell suspension is sub-packed in 5cm
2Culture bottle places in the incubator and cultivates.Changed in second day and adopt the M199 culture fluid that contains 30ug/ml ECGS, changed a culture fluid and be paved with the bottom surface in per afterwards three days until cell.Used cell is former generation endotheliocyte.The culture bottle inner cell is seeded to 96 orifice plates before the modeling and cultivated one day, used cell concentration is 3.5～40,000/hole.During modeling, inhale and abandon old M199 culture fluid, adding pastille does not have phenol red M199 culture fluid, and every hole total amount is 100ul.Place 95%N2 and 5%CO
2Anoxia is 12 hours in the incubator, then at CO
2Reoxygenation is 2 hours in the incubator.Get cell conditioned medium liquid and measure LDH content and NO content.
Dosage regimenThe Danshen effective component that extraction obtains is made into 10 with sugar-free Hanks liquid
-4G/mL test liquid, liposoluble constituent can add a small amount of DMSO hydrotropy (the DMSO final concentration is controlled at below 0.2%).The final concentration of each component in no phenol red M199 culture fluid is controlled at 10
-5G/mL.
The NO assayAdopt Greiss reagent method to measure, get 90ul cell conditioned medium liquid, add equivalent Greiss reagent, left standstill under the room temperature 10 minutes, measure absorbance in 550nm with microplate reader.
Drug effect resultAll data are represented with the mean value variance.Adopt monolateral variance analysis and T check carrying out statistical analysis.Compare with model group, P＜0.05 is thought significantly effectively.Drug effect the results are shown in Table 2.According to endotheliocyte NO content detection result, the Fructus Aurantii active component has the highly significant effect to NO overexpression after suppressing endotheliocyte anoxia-reoxygenation.
Table 2
Claims (6)
1. Danshen effective component, it is characterized in that, by weight percentage, the content of dihydrotanshinone I is 96.5～98%, adopts following method to prepare: get red rooted salvia, it is pulverized the back add ethyl acetate and ethanol, reflux 1 hour, extract 2 times, merging filtrate gets extracting solution, and extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate, get eluent I as mobile phase, abandon it, change chloroform and methanol then as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is semi-preparative column LichrospherC18; 10.0mm * 250mm, 5 μ m, mobile phase is water A and acetonitrile B, the gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 50% aqueous solution, Mobile phase B are 50% acetonitrile solution; In the time of 35 minutes, mobile phase A is that 20% aqueous solution, Mobile phase B are 80% acetonitrile solution; In the time of 45 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; In the time of 55 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; Flow velocity is 3ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 21.7～30.3min, and solution obtains active component behind concentrate drying; Wherein ethyl acetate and alcoholic acid volume ratio are 1: 1, and the volume ratio of petroleum ether and ethyl acetate 50: 1, the volume ratio of chloroform and methanol are 10: 1.
2. Danshen effective component as claimed in claim 1 is characterized in that, by weight percentage, the content of dihydrotanshinone I is 96.8～97.7%.
3. Danshen effective component as claimed in claim 1 is characterized in that, by weight percentage, the content of dihydrotanshinone I is 97.0～97.5%.
4. the preparation method of Danshen effective component comprises the following steps: to get red rooted salvia, it is pulverized the back add ethyl acetate and ethanol, reflux 1 hour is extracted 2 times, and merging filtrate gets extracting solution, extracting solution is condensed into extractum, and itself and silica gel are mixed sample, it is separated with normal phase silicagel column, at first use petroleum ether and ethyl acetate as mobile phase, get eluent I, abandon it, change chloroform and methanol then as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is semi-preparative column Lichrospher C18; 10.0mm * 250mm, 5 μ m, mobile phase is water A and acetonitrile B, the gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 50% aqueous solution, Mobile phase B are 50% acetonitrile solution; In the time of 35 minutes, mobile phase A is that 20% aqueous solution, Mobile phase B are 80% acetonitrile solution; In the time of 45 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; In the time of 55 minutes, mobile phase A is that 5% aqueous solution, Mobile phase B are 95% acetonitrile solution; Flow velocity is 3ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 21.7～30.3min, and solution obtains active component behind concentrate drying; Wherein ethyl acetate and alcoholic acid volume ratio are 1: 1, and the volume ratio of petroleum ether and ethyl acetate 50: 1, the volume ratio of chloroform and methanol are 10: 1.
5. contain the pharmaceutical formulation of the described Danshen effective component of the arbitrary claim of claim 1-3, it is characterized in that, make arbitrary pharmaceutical formulation that pharmaceutics is accepted according to the preparation method that pharmaceutics allows.
6. the application of the described Danshen effective component of the arbitrary claim of claim 1-3 in preparation treatment and angiocardiopathy preventing medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2005101223489A CN1981811B (en) | 2005-12-14 | 2005-12-14 | Danshen effective component, its preparation, making method and usage |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2005101223489A CN1981811B (en) | 2005-12-14 | 2005-12-14 | Danshen effective component, its preparation, making method and usage |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1981811A CN1981811A (en) | 2007-06-20 |
| CN1981811B true CN1981811B (en) | 2010-12-15 |
Family
ID=38164841
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2005101223489A Expired - Fee Related CN1981811B (en) | 2005-12-14 | 2005-12-14 | Danshen effective component, its preparation, making method and usage |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1981811B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1394870A (en) * | 2001-07-05 | 2003-02-05 | 北京天纯维通生物技术有限公司 | Method for separating and purifying tanshinone |
| CN1670019A (en) * | 2004-03-17 | 2005-09-21 | 天津天士力现代中药资源有限公司 | Process for extracting tanshinone |
-
2005
- 2005-12-14 CN CN2005101223489A patent/CN1981811B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1394870A (en) * | 2001-07-05 | 2003-02-05 | 北京天纯维通生物技术有限公司 | Method for separating and purifying tanshinone |
| CN1670019A (en) * | 2004-03-17 | 2005-09-21 | 天津天士力现代中药资源有限公司 | Process for extracting tanshinone |
Non-Patent Citations (2)
| Title |
|---|
| DONG-SUN LEE,SANG-HAN LEE.Biological Activity of Dihydrotanshinone I: Effect on Apoptosis.Journal of Bioscience and Bioengineering.2000,89(3),292-293. * |
| 杨勤,赵朝伟.丹参的药理作用研究现状.中国药业.2003,12(10),78-79. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1981811A (en) | 2007-06-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102362971B (en) | A kind of preparation method and preparation for the treatment of coronary heart disease Chinese medicine and effective chemical constituent thereof | |
| CN101874841A (en) | Total glycosides extractive of morinda plants, as well as preparation method and application thereof | |
| CN102920964A (en) | Traditional Chinese medicine preparation for curing cough | |
| CN1989984B (en) | Chuanxiong rhizome effective ingredient, preparing method, preparation and use thereof | |
| CN1981810B (en) | Danshen component, preparation, making method and usage | |
| CN1981809B (en) | Danshen effective component, its preparation, making method and usage | |
| CN1981811B (en) | Danshen effective component, its preparation, making method and usage | |
| CN101785799B (en) | Extract of total triterpenes in dracocephalum moldavica and preparation method and application thereof | |
| CN102100737A (en) | Medicinal composition containing general ginsenoside and total salvianolic acid and preparation method thereof | |
| CN103977336A (en) | Pharmaceutical composition with myocardial ischemia resisting effect and preparation method and application thereof | |
| CN1887324B (en) | Chinese medicine composition for treating liver and kidney defect, and its preparation process and analysis method | |
| CN113577123A (en) | Preparation method and application of dandelion extract | |
| CN1989985B (en) | Chuanxiong rhizome effective ingredient, preparing method, preparation and use thereof | |
| CN1981822B (en) | Tree-peony root bark effective component, its preparation, making method and use | |
| CN1981819B (en) | Tree-peony root bark component, its preparation, making method and use | |
| Rahhal et al. | Phytochemical investigation and diuretic activity of the Palestinian rataegus aronia in mice using an aqueous extract | |
| CN1989986B (en) | Chuanxiong rhizome effective ingredient, preparing, preparation method and use thereof | |
| CN102125590B (en) | Ginsenoside Rg1-containing pseudo-ginseng active ingredient as well as preparation method and application thereof | |
| CN101474264A (en) | Effective component of white peony root as well as preparation method and use thereof | |
| CN100427115C (en) | Effective part and preparation method of traditional Chinese medicine paeonol for treating cardiovascular disease | |
| CN1981790B (en) | Rhizoma Chuanxiong effective components and production thereof | |
| CN1981821A (en) | Tree-peony root bark effective component, its preparation, making method and use | |
| CN1981820A (en) | Tree-peony root bark effective component, its preparation, making method and use | |
| CN102133262A (en) | Preparation method and application of Astragalus membranaceus active constituent with effect of myocardium protection | |
| CN1981818B (en) | Chinese-medicinal composition, preparation and use for treating cardiovascular diseases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| PB01 | Publication | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20070511 Address after: Hedong District of Beichen Puji road in Tianjin No. 2 city of the modern Chinese Medicine Applicant after: Tianjin Tianshili Pharmaceutical Co., Ltd. Address before: White road, Beichen science and Technology Park in Tianjin City Xinyi Liaohe Road No. 1 Applicant before: Tianshili Modern Traditional Chinese Medicine Research & Devleopment Co., Ltd., |
|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: TASLY PHARMACEUTICAL GROUP CO., LTD. Free format text: FORMER NAME: TIANJIN TASLY PHARMACEUTICAL CO., LTD. |
|
| CP01 | Change in the name or title of a patent holder |
Address after: 300402 Tianjin City Hedong District of Beichen Puji Road No. 2 city of the modern Chinese Medicine Patentee after: Tasly Pharmaceutical Group Co., Ltd. Address before: 300402 Tianjin City Hedong District of Beichen Puji Road No. 2 city of the modern Chinese Medicine Patentee before: Tianjin Tianshili Pharmaceutical Co., Ltd. |
|
| CP01 | Change in the name or title of a patent holder |
Address after: 300402 Tianjin City Hedong District of Beichen Puji Road No. 2 city of the modern Chinese Medicine Patentee after: Tasly Pharmaceutical Group Limited by Share Ltd Address before: 300402 Tianjin City Hedong District of Beichen Puji Road No. 2 city of the modern Chinese Medicine Patentee before: Tasly Pharmaceutical Group Co., Ltd. |
|
| CP01 | Change in the name or title of a patent holder | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20101215 Termination date: 20201214 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |