CN102145046B - Preparation method and applications of gallicin-containing red peony root active component - Google Patents
Preparation method and applications of gallicin-containing red peony root active component Download PDFInfo
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- CN102145046B CN102145046B CN2011100576370A CN201110057637A CN102145046B CN 102145046 B CN102145046 B CN 102145046B CN 2011100576370 A CN2011100576370 A CN 2011100576370A CN 201110057637 A CN201110057637 A CN 201110057637A CN 102145046 B CN102145046 B CN 102145046B
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- paeoniae rubra
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Abstract
The invention provides a gallicin-containing red peony root active component, which is prepared by the following steps: extracting under heating, concentrating, separating with a reverse phase silica gel column, eluting, concentrating and drying an eluent, separating with a liquid chromatography, collecting a solution, and concentrating and drying to obtain the active component. The active component mainly comprises gallicin and pentagalloylglucose. The red peony root active component provided by the invention can be used in preparing a medicine for treating and preventing cardiovascular diseases. The method provided by the invention is reasonably designed, can rapidly and accurately obtain the active component, and is more favorable for quality control of the medicine in production.
Description
Technical field
The invention belongs to the field of Chinese medicines; Relate to a kind of Chinese medicine extract of treating cardiovascular disease; Relate in particular to the method for preparing and the pharmaceutical applications thereof of a kind of Radix Paeoniae Rubra active component of from Radix Paeoniae Rubra extraction, this active component mainly contains gallicin, five galloyl glucoses.
Background technology
Cardiovascular disease is the first healthy killer of harm humans; In recent years; Along with the variation of China's aged tendency of population 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.Many active substances have resisting myocardial ischemia, anoxia functions in the natural product, and some of them have been developed to treatment coronary heart disease and anginal new drug.Thereby from natural product, seek active substance with resisting myocardial ischemia, anoxia physiologically active, be one of effective way of discovery, 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 and have resisting myocardial ischemia; Be used to treat coronary heart disease and anginal new drug, have significant application value and wide development prospect.
Radix Paeoniae Rubra be the Ranunculaceae Radix Paeoniae (
Paeonia lactifloraPall.) or river Radix Paeoniae Rubra (
P.veitchii Lynch.) dry root.Its bitter in the mouth is slightly cold, and returns Liver Channel, has clearing away heat and cooling blood, the effect of dissipating blood stasis analgesia.Modern study shows; Mainly contain glycosides compounds such as peoniflorin, Hydroxy peoniflorin, benzoylpaeoniflorin, oxypaeoniflorin, peonin, galloylpaeoniflorin in the Radix Paeoniae Rubra, also have materials such as iridoids, triterpenes, phenol and glycoside, sterol and glycoside thereof, tannin, acids, esters, flavonoid, ketone, alkanes, volatile oil, sugar, aminoacid, protein, fatty oil, resin, lymphatic temperament, alkaloid, pigment, trace element in addition.
Radix Paeoniae Rubra is traditional blood-activating stasis-removing kind Chinese medicine, and stronger study of anti-atherogenic effect is arranged, and this medicine also can dilating coronary blood vessel; Thereby coronary blood flow increasing increases myocardial nutrition property blood flow, and the protection ischemic myocardium improves the toleration of cardiac muscle to hypoxia; Reduce pulmonary vascular resistance, alleviate afterload.Radix Paeoniae Rubra can reduce portal hypertension simultaneously, and is also influential to microcirculation.Pharmaceutical research shows Radix Paeoniae Rubra ability anticoagulant, improves erythrocytic permeability, increases erythrocyte to hypotonic tensile resistance, and the effect of necessarily stablizing the erythrocyte membrane structure is arranged.In addition, the clotting time of obvious prolong rats of Radix Paeoniae Rubra total glycosides ability and mice, and can prolong the thrombotic time of electricity irritation rat carotid artery, the prompting Radix Paeoniae Rubra total glycosides produces anti thrombotic action through the influence to blood coagulation system and platelet function.Radix Paeoniae Rubra has above obvious pharmacological action to cardiovascular system, therefore, from Radix Paeoniae Rubra, isolates active component, and 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 method for preparing that contains the Radix Paeoniae Rubra active component of gallicin, realize: with adding ethyl acetate and ethanol, heating extraction after the Radix Paeoniae Rubra pulverizing medicinal materials through following steps; Medicinal residues add ethanol water and extract, and extracting solution is condensed into extractum, with the reverse phase silica gel post it are separated; At first use low concentration methanol as mobile phase; Get eluent I, abandon it, use high concentration methanol then instead as mobile phase; Get eluent II, with getting 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, and gradient elution mainly contains gallicin, five galloyl glucoses in this active component.
Preferred manufacturing procedure of the present invention, realize through following steps: get the Radix Paeoniae Rubra medical material, it is pulverized the back, and to add volume ratio be ethyl acetate and the ethanol of 1:1, and reflux 1 hour is extracted 2 times, filtrate merge extracting solution I.Medicinal residues add 70% ethanol, and reflux 1 hour is extracted 2 times, filtrating merge extracting solution II, II is condensed into extractum with extracting solution, with 50% dissolve with methanol extractum, the employing ODS-C
18Post separates it, at first uses 5% methanol as mobile phase, gets eluent I, uses 60% methanol then instead as mobile phase, gets eluent II, gets 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 C
18; 10.0mm
250mm, 5
M), mobile phase is water A and acetonitrile B, and the gradient elution program is following: during 0min, mobile phase A is 90% water, and Mobile phase B is 10% acetonitrile solution; During 15min, mobile phase A is 75% water, and Mobile phase B is 25% acetonitrile solution; During 34min, mobile phase A is 70% water, and Mobile phase B is 30% acetonitrile solution; Flow velocity is 3ml/min, and column temperature is a room temperature; Sample separates through preparative liquid chromatography, collects solution at time period 26.8 ~ 31.0min, and solution obtains active component behind concentrate drying, mainly contain gallicin, five galloyl glucoses in this active component.
Another object of the present invention provides the application of said Radix Paeoniae Rubra active component in the medicine of preparation treatment, angiocardiopathy preventing.Prove that through pharmacological evaluation Radix Paeoniae Rubra active component provided by the invention has protective effect to myocardial cell.
Radix Paeoniae Rubra active component of the present invention can be used as active component, adds the adjuvant of accepting on the pharmaceutics, processes preparation according to the method for preparing 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.
Radix Paeoniae Rubra active component provided by the invention can be in the application in preparation treatment, the angiocardiopathy preventing medicine.Method for preparing provided by the invention is reasonable in design, can obtain effective ingredient rapidly and accurately, is easier to the quality control of medicine aborning.
Description of drawings
Fig. 1 is the HPLC chromatogram of Radix Paeoniae Rubra active component.
Fig. 2 is No. 1 chromatographic peak mass spectrum.
Fig. 3 is No. 2 chromatographic peak mass spectruies.
The specific embodiment
To combine embodiment further explain flesh and blood of the present invention and beneficial effect below, this embodiment only is used to the present invention is described but not limitation of the present invention.
The preparation of embodiment one Radix Paeoniae Rubra active component
With adding ethyl acetate and ethanol after the Radix Paeoniae Rubra pulverizing medicinal materials, heating extraction gets extracting solution I, abandons it.Medicinal residues add ethanol water, and heating extraction gets extracting solution II, and II is condensed into extractum with extracting solution, with 50% dissolve with methanol extractum, adopt ODS-C
18Post separates it, at first uses low concentration methanol as mobile phase, gets eluent I, uses high concentration methanol then instead as mobile phase, gets eluent II, gets sample behind the concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography, separate, collect solution, obtain active component behind the concentrate drying in 26.8 ~ 31.0 time periods through preparative liquid chromatography.
The preparation of embodiment two Radix Paeoniae Rubra active components
Get Radix Paeoniae Rubra medical material 250g, it is pulverized the back, and to add volume ratio be ethyl acetate and the ethanol of 1:1, and reflux 1 hour is extracted 2 times, filtrate merge extracting solution I.Medicinal residues add 70% ethanol, and reflux 1 hour is extracted 2 times, filtrating merge extracting solution II, II is condensed into extractum with extracting solution, with 50% dissolve with methanol extractum, the employing ODS-C
18Post separates it, at first uses 5% methanol as mobile phase, gets eluent I, uses 60% methanol then instead as mobile phase, gets eluent II, gets 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 C
18; 10.0mm
250mm, 5
), mobile phase is water A and acetonitrile B, the gradient elution program is following: during 0min, mobile phase A is 90% water, and Mobile phase B is 10% acetonitrile solution; During 15min, mobile phase A is 75% water, and Mobile phase B is 25% acetonitrile solution; During 34min, mobile phase A is 70% water, and Mobile phase B is 30% acetonitrile solution; Flow velocity is 3ml/min, and column temperature is a room temperature; Sample separates through preparative liquid chromatography, collects solution at time period 26.8 ~ 31.0min, and solution obtains active component behind concentrate drying.This active component mainly contains gallicin, five galloyl glucoses, and its chromatogram and mass spectrum are seen Fig. 1 ~ 3.
The preparation of embodiment three dropping pill formulations
Get Radix Paeoniae Rubra active component 0.5g and 10.5g Polyethylene Glycol-20000 mix homogeneously, 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 400 of drop pill.
The preparation of embodiment four lyophilized injectable powders
Get Radix Paeoniae Rubra active component 0.5g, glucose 4.5g, sodium thiosulfate 0.9g and distilled water 1000ml, behind the said components mix homogeneously, 400 of packing, lyophilization promptly gets.
The active appraisal experiment of embodiment five Radix Paeoniae Rubra components
1. the Radix Paeoniae Rubra component is to H
2O
2The protective effect of injury of myocardium cell
In vitro culture H
9C
2Myocardial cell, culture fluid are that DMEM (high sugar)+10%FBS (G)+1% non essential amino acid+0.1% pair is anti-.The trophophase cell of taking the logarithm places on the 96 porocyte culture plates, behind constant temperature culture 24 h, adds 200 μ mol/mL H
2O
2Injury of myocardium cell 30 min, the Radix Paeoniae Rubra component that adds final concentration again and be 50 μ g/mL is hatched 24 h, adopts mtt assay to measure cell viability.The result shows that this component is to H
2O
2The protective rate of injury of myocardium cell is 14.95%.
2. the Radix Paeoniae Rubra component is to the protective effect of hypoxia-reoxygenation induced injured myocardium cell
In vitro culture H
9C
2Myocardial cell, culture fluid are that DMEM (high sugar)+10%FBS (G)+1% non essential amino acid+0.1% pair is anti-.The trophophase cell of taking the logarithm places on the 96 porocyte culture plates, behind constant temperature culture 24 h, in the anoxia cell, cultivates 6 h.After anoxia finished, taking out the Tissue Culture Plate adding was the Radix Paeoniae Rubra component of 50 μ g/mL through the final concentration of sugar-free Hank ' s dilution, under the normal cultured condition, hatches 6 h, gets cell culture supernatant and measures lactic acid dehydrogenase (LDH) content.The result shows that this component is 44.20% to the myocardial cell protection rate of anoxia reoxygenation injury.
Claims (4)
1. a Radix Paeoniae Rubra active component is characterized in that, obtains through following steps: get the Radix Paeoniae Rubra medical material, it is pulverized the back, and to add volume ratio be 1: 1 ethyl acetate and ethanol, and reflux 1 hour is extracted 2 times, filtrate merge extracting solution I; Medicinal residues add 70% alcoholic solution, and reflux 1 hour is extracted 2 times, filtrating merge extracting solution II, II is condensed into extractum with extracting solution, with 50% dissolve with methanol solution extractum, the employing ODS-C
18Post separates it, at first uses 5% methanol solution as mobile phase, gets eluent I, uses 60% methanol solution then instead as mobile phase, gets eluent II, gets sample 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, Lichrospher C
1810.0mm * 250mm, 5 μ m, mobile phase is water A and acetonitrile B, the gradient elution program is following: 0min, 10%B; 15min, 25%B; 34min, 30%B; Flow velocity is 3ml/min, and column temperature is a room temperature; Sample separates through preparative liquid chromatography, collects solution in 26.8～31.0min time period, and solution obtains active component behind concentrate drying; Mainly contain gallicin, five galloyl glucoses in this active component.
2. the application of a kind of Radix Paeoniae Rubra active component according to claim 1 in the medicine of preparation treatment cardiovascular disease.
3. application according to claim 2 is characterized in that, said medicine adds acceptable auxiliary on the pharmaceutics by the Radix Paeoniae Rubra active component, processes according to the formulation preparation method of putting down in writing on the pharmaceutics.
4. application according to claim 3 is characterized in that described medicine comprises liquid preparation or solid preparation.
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| CN2011100576370A CN102145046B (en) | 2011-03-10 | 2011-03-10 | Preparation method and applications of gallicin-containing red peony root active component |
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| CN2011100576370A CN102145046B (en) | 2011-03-10 | 2011-03-10 | Preparation method and applications of gallicin-containing red peony root active component |
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| CN102145046B true CN102145046B (en) | 2012-06-27 |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101347518A (en) * | 2007-07-20 | 2009-01-21 | 天津天士力制药股份有限公司 | Effective component of red paeonia and preparation and use thereof |
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2011
- 2011-03-10 CN CN2011100576370A patent/CN102145046B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101347518A (en) * | 2007-07-20 | 2009-01-21 | 天津天士力制药股份有限公司 | Effective component of red paeonia and preparation and use thereof |
Non-Patent Citations (1)
| Title |
|---|
| 刘虹等.大孔树脂纯化赤芍提取物的工艺考察.《天津中医药大学学报》.2009,第28卷(第3期),第142-145页. * |
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