CN101696166B - Preparation method for danshen root salvianolic acid A - Google Patents
Preparation method for danshen root salvianolic acid A Download PDFInfo
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- CN101696166B CN101696166B CN2009101699008A CN200910169900A CN101696166B CN 101696166 B CN101696166 B CN 101696166B CN 2009101699008 A CN2009101699008 A CN 2009101699008A CN 200910169900 A CN200910169900 A CN 200910169900A CN 101696166 B CN101696166 B CN 101696166B
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- China
- Prior art keywords
- salvianolic acid
- extract
- danshen root
- preparation
- root salvianolic
- Prior art date
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- YMGFTDKNIWPMGF-AGYDPFETSA-N 3-(3,4-dihydroxyphenyl)-2-[(e)-3-[2-[(e)-2-(3,4-dihydroxyphenyl)ethenyl]-3,4-dihydroxyphenyl]prop-2-enoyl]oxypropanoic acid Chemical compound C=1C=C(O)C(O)=C(\C=C\C=2C=C(O)C(O)=CC=2)C=1/C=C/C(=O)OC(C(=O)O)CC1=CC=C(O)C(O)=C1 YMGFTDKNIWPMGF-AGYDPFETSA-N 0.000 title claims abstract description 84
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- Medicinal Preparation (AREA)
Abstract
The invention discloses a preparation method for danshen root salvianolic acid A, which is characterized in that total phenolic acid in danshen root is converted into the salvianolic acid A by using a certain method so as to greatly improve the extraction efficiency of the salvianolic acid A. The danshen root salvianolic acid A is prepared into tablets, capsules, granules, soft capsules, pellets, pills and oral liquid. Pharmacological experiments show that the salvianolic acid A has good pharmacological effects.
Description
The present invention is that application number is 200710001055.4 divides an application, and the applying date of original bill is on January 23rd, 2007, and application number is 200710001055.4, and denomination of invention is: a kind of preparation method of danshen root salvianolic acid A.
Technical field
The present invention relates to medicine, health food technology field, be specifically related to a kind of preparation method of danshen root salvianolic acid A.
Background technology
The red sage root is the root of the Labiatae salvia red sage root.Bitter, cold nature; The red sage root is one of the most frequently used medicinal material; Being used to treat cardiovascular and cerebrovascular diseases such as stenocardia, hypertension, coronary heart disease and apoplexy, having the effect of promoting blood circulation and removing blood stasis, nourishing blood to tranquillize the mind, cool blood row's carbuncle and toxin expelling myogenic, is Chinese medicine flavour of a drug commonly used promoting blood circulation and removing blood stasis.
The chemical ingredients of the red sage root mainly can be divided into water soluble component and fat-soluble component; Red sage root water soluble ingredient has phenolic acid property structure more; Be effective constituents such as salvianolic acid A, B, C, D, E, the highest in the Radix Salviae Miltiorrhizae total phenolic acids with content of danshinolic acid B, therefore; The emphasis of people's exploitation concentrates on salvianolic acid B, and content is lower, active stronger salvianolic acid A (salvianolic acid A) is ignored by people because can't carry out suitability for industrialized production.Consult document, we can learn therefore the content of salvianolic acid A in the red sage root is about 5/10000ths; Even it is extracted through a series of method; Can only obtain the trace danshen root salvianolic acid A, be applied to clinical in, can not be accepted by the patient because its price is too high; But danshen root salvianolic acid A has good pharmacologically active, can be applied in medicine, the protective foods, and therefore, a lot of medical workers hope salvianolic acid A is carried out suitability for industrialized production, are developed as the patent medicine preparation.
The process method of danshen root salvianolic acid A extraction purifying is disclosed in existing document and the patent; Though can access danshen root salvianolic acid A; But how its emphasis all is placed on danshen root salvianolic acid A and the separate impurities, and its extraction yield is very low, can't in suitability for industrialized production, realize; Also can't obtain the danshen root salvianolic acid A of " patent medicine " level, this is one difficulty of pendulum in face of a lot of researchers.
Summary of the invention
For these reasons, we find through long term studies, after Radix Salviae Miltiorrhizae total phenolic acids is extracted; Under certain condition; Can Radix Salviae Miltiorrhizae total phenolic acids be changed into danshen root salvianolic acid A, thereby improve the extraction yield of danshen root salvianolic acid A greatly, its extraction yield can reach 1%-2%; On the basis that has solved extraction yield, the formulation preparation of salvianolic acid A also is readily solved.
The invention reside in provides a kind of process method that improves the danshen root salvianolic acid A extraction yield;
The present invention also is to provide a kind of preparation method who contains the danshen root salvianolic acid A preparation.
The present invention realizes through following technical scheme.
One. process method
Danshen root salvianolic acid A extracts:
Method 1: red sage root water or ethanolic soln extract and obtain aqueous extract or alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 3.5-6.0,110-130 ℃ temperature; Heated 1-6 hour; Solution filters, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 2: red sage root water or ethanolic soln extract and obtain aqueous extract or alcohol extract; Alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 3.5-6.0, puts into the microwave extraction device; Frequency is that 915MHz-2450MHz, power are 1000-15000 watt microwave extraction 0.5-2 hour; Extracting liquid filtering, the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 3: red sage root water or ethanolic soln extract and obtain aqueous extract or alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 3.5-6.0,110-130 ℃ temperature, heats 1-6 hour; Solution filters, and filtrating is separated first water, 10-30% Diluted Alcohol wash-out through nonpolar or low-pole macroporous resin column chromatography; Remove impurity, use the ethanol elution of 30-70% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Method 4: red sage root water or ethanolic soln extract and obtain aqueous extract or alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 3.5-6.0; Put into the microwave extraction device, frequency is that 915MHz-2450MHz, power are 1000-15000 watt microwave extraction 0.5-2 hour, extracting liquid filtering; Filtrating is separated through nonpolar or low-pole macroporous resin column chromatography, and first water, 10-30% Diluted Alcohol wash-out are removed impurity; Use the ethanol elution of 30-70% concentration again, collect elutriant, reclaim ethanol to most; Drying obtains the danshen root salvianolic acid A extract;
Wherein said macroporous resin column is HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8.
Contain the pharmaceutical composition that the danshen root salvianolic acid A extract is an activeconstituents.
Formulation preparation: get above-mentioned danshen root salvianolic acid A, learn the conventional preparation that requires to be prepared into according to tablet, capsule, granule, soft capsule, pellet, pill, oral liquid medicine.
The solid preparation of the application's preparation carries out the dissulution experiment, and dissulution reaches more than 80% in the time of 45 minutes.
Two. detection method
Experimental technique:
Chromatographic column: C
18Reverse-phase chromatographic column, NUCLEODUR, 250*4.6mm, ODS;
Chromatographic condition and system suitability experiment: with the octadecylsilane chemically bonded silica is weighting agent; Flow velocity 1.0ml/min; 35 ℃ of column temperatures; Detect wavelength 286nm; Number of theoretical plate should be not less than 60000 by salvianolic acid A; With acetonitrile-0.2% aqueous acetic acid is moving phase, carries out gradient elution by following condition of gradient elution, moves 90 minutes;
In the time of 0-15 minute, the ratio of acetonitrile by 10% rise to 20%, 0.2% aqueous acetic acid ratio reduce to 80% by 90%; In the time of 15-55 minute, the ratio of acetonitrile by 20% rise to 30%, 0.2% aqueous acetic acid ratio reduce to 70% by 80%; In the time of 55-65 minute, the ratio of acetonitrile by 30% rise to 50%, 0.2% aqueous acetic acid ratio reduce to 50% by 70%; In the time of 65-72 minute, the ratio of acetonitrile by 50% rise to 80%, 0.2% aqueous acetic acid ratio reduce to 20% by 50%; In the time of 72-77 minute, the ratio 20% of ratio 80%, 0.2% aqueous acetic acid of acetonitrile; In the time of 77-80 minute, the ratio of acetonitrile by 80% reduce to 10%, 0.2% aqueous acetic acid ratio rise to 90% by 20%; In the time of 80-90 minute, keep acetonitrile-0.2% aqueous acetic acid to carry out wash-out with 10: 90 ratio;
The preparation of reference substance solution: precision takes by weighing the salvianolic acid A reference substance in volumetric flask, adds dissolve with methanol and shakes up, and be diluted to scale;
The preparation of sample solution: sample thief adds dissolve with methanol and shakes up; Or precision measures or takes by weighing preparation, adds dissolve with methanol and shakes up, and be diluted to scale;
Assay method: the accurate reference substance solution of drawing, inject liquid chromatograph, the record color atlas; The accurate sample solution of drawing injects liquid chromatograph, calculates peak area ratio.
Three. the extraction yield experiment
Experimental technique: get red rooted salvia,, measure danshen root salvianolic acid A content according to above-mentioned check and analysis experimental technique; Be divided into six equal portions, portion method (Chinese patent CN1397276A, scheme 1) traditionally obtains the danshen root salvianolic acid A extract; A according to process method (Chinese patent CN1830947; Scheme 2), other four parts obtain the danshen root salvianolic acid A extract according to the application's process method (being followed successively by scheme 3-6) respectively, according to salvianolic acid A weight in the above-mentioned check and analysis experimental calculation danshen root salvianolic acid A extract; According to the extraction yield of salvianolic acid A weight calculating different process method, experimental result is seen table 1:
The extraction yield of table 1 different process method salvianolic acid A
Experiment conclusion: show through above-mentioned experiment; The extraction yield that the application's process method obtains salvianolic acid A has improved more than 100 times than the extraction yield of salvianolic acid A in the prior art; This can't realize through simple extraction and purification process; Must Radix Salviae Miltiorrhizae total phenolic acids be changed into salvianolic acid A through certain conversion, could fundamentally improve the extraction yield of salvianolic acid A.
Four. pharmacological evaluation
Experiment 1
1. to the provide protection of anesthetized rat myocardial ischemia-reperfusion injury
Experimental technique:
Get the healthy SD rat, body weight 240-260g, random packet: blank group, FUFANG DANSHEN PIAN group, the application's solid preparation of salvianolic acid A of red sage root group.Place equivalent environment to raise 2 days free diet in advance.After raising end in advance, make an experiment, animal is weighed, and 20% urethane is pressed the 0.6ml/100g abdominal injection; After treating that anesthesia is satisfied, lie on the back and be fixed on the mouse plate, trachea cannula connects respirator; By 10～12ml Tidal volume, 70 times/minute frequency is exhaled, and continuous positive pressure breathing is inhaled: exhale than being 1: 1.According to respiratory rate and degree of depth adjustment respiration parameter.Connect electrocardiograph subsequently, survey normal ECG.Cut off front field of operation hair, iodine disinfection is cut off skin, subcutis, front muscle and manadesma 3～4cm; It is long to separate intercostal muscle 3cm with the 18# vascular clamp along the 3rd intercostal passivity, opens thoracic cavity and pericardium, recording ecg; Strut 3,4 ribs, refer to hold thoracic cavity, rat right side with left hand four, the assistant upwards pushes away thymus gland with the ophthalmology tweezer; Between left auricle of heart and pulmonary conus, find ligation sign blood vessel great cardiac vein, the 2mm place is not with there being wound roundlet pin band 6-0 silk thread threading below left auricle of heart, and depth of needle is 1～1.5mm; Wide 2～3mm, recording ecg behind the threading gives corresponding soup through the tail vein; Recording ecg behind the administration 10min, and with one the band groove little plastics pipe pad at the ligation position, the ligation above that of two rear line heads.At once recording ecg after the ligation is cyanosis or the II S-T section back of a bow that leads with left chamber antetheca and upwards raises greater than 0.1mv and be that ligation successfully indicates (it is superseded that the S-T section does not have the changer) more than the lasting 0.5h.10min recording ecg is once more cut off ligature behind the ligation 30min after the ligation, realizes perfusion again, and record pours into electrocardiogram(ECG at once again, removes in the thoracic cavity layer-by-layer suture wall of the chest behind the hematocele, removes respirator, animal recovery autonomous respiration, and incision of trachea does not process.Irritate again at once, 10min, 20min, 40min, 1h, 2h, 3h recording ecg respectively.With heart after refrigerator and cooled is frozen 10min, from the apex of the heart entad the parallel coronary sulcus direction in the end 5 of equal thickness are cut in left chamber, put into the 1%TTC dye liquor, 37 ℃ of dyeing 10min, the necrotic area is not a garnet, the necrotic area is pearl.Digital camera is taken pictures.Weighed respectively in necrotic area and non-necrotic area, calculate the per-cent that the necrotic area accounts for left ventricular mass, i.e. infarction size.
Table 4 solid preparation of salvianolic acid A is to the influence of myocardial ischemia myocardial infarct size (%) due to the logical rat ramus descendens anterior arteriae coronariae sinistrae of ligation/again (x ± s)
Annotate: compare with the blank group,
*P＜0.01; Compare #P＜0.05 with positive FUFANG DANSHEN PIAN group
Experiment 2
Research to the provide protection of intraluminal middle cerebral artery occlusion in rats ischemical reperfusion injury
Experimental technique:
Animal random packet: blank group, FUFANG DANSHEN PIAN group, the application's danshen root salvianolic acid A preparation group.The continuous gastric infusion of each dose groups 3 days was processed middle cerebral artery occlusion (MCAO) model with improvement line bolt method in 20 minutes behind the 4th day medicine.Behind the rat anesthesia, it is fixing that it is lain on the back.Separate right carotid (CCA), internal carotid artery (ICA) and external carotid artery (ECA), ligation ECA and CCA, close the ICA distal end with the bulldog clamp folder after; Make a kerf in ECA and ICA crotch rapidly; Insert the nylon wire (diameter is 0.25mm, and 18mm marks at the place apart from the pommel, is stained with heparin solution before the insertion) that an end is heated into smooth, spherical and has been coated with 0.1% poly-lysine; Depth of penetration is 18mm, realizes that middle cerebral artery occlusion causes cerebral ischemia.Ligation ingress, nylon wire are stayed about 1cm, skin suture outward.Lift extremely slightly resistance of institute's the end of a thread that stays after 2 hours gently, realize that arteria cerebri media pours into again, modeling is accomplished.At ischemic 2h with pour in the 1h body temperature of keeping rat with electric blanket again, body temperature maintains 36.5～37.5 ℃ of anus temperature.The animal inclusion criteria is pressed Longa Pyatyi point system, gets the neural function behavior scoring and be 1,2,3,4 minute animal, (0 minute: the impassivity defective symptom; 1 minute: the offside forelimb can not stretch fully; 2 minutes: to sideway swivel; 3 minutes: topple over to offside; 4 minutes: can not oneself walk or stupor).The cerebral infarction scope is measured, and rat model pours into 24h again, after the study of behaviour scoring; Broken end is got brain, removes olfactory bulb, cerebellum and low brain stem, and remainder is at-20 ℃ of freezing 10min of refrigerator; Crownly on ice pan be cut into 6; Rapidly the brain sheet is placed the TTC dye liquor, 37 ℃ of lucifuge temperature are incubated 1h, take out to be placed on the 24h that keeps in Dark Place in 10% formalin.The non-ischemic region in dyed back is a rose, and infarct is a white.White organized carefully to dig down weigh, account for full brain weight per-cent as the cerebral infarction scope with blocking tissue's weight.
Brain water content is measured: after TTC dyeing is weighed, brain placed oven dry 12h claims dry weight in 120 ℃ of vacuum driers.Brain water content=(brain weight in wet base-brain stem is heavy)/brain weight in wet base * 100%.Experimental result sees table 5 for details.
The influence of table 5 pair intraluminal middle cerebral artery occlusion in rats ischemical reperfusion injury rat cerebral infarction scope and brain water content (X ± S)
Annotate: compare with the blank group,
*P＜0.01; Compare #P＜0.05 with positive FUFANG DANSHEN PIAN group
Experiment 3
To CCl
4Cause the influence of rat liver fibrosis
Experimental technique: select 60 of male and healthy Wistar rats; Body weight 180-200g; All divide normal group, NSC-757. group (0.17mg/kg), the application's danshen root salvianolic acid A preparation group at random with rat, except that the normal control group, all the other respectively organize rat first in subcutaneous injection CCl
45ml/kg, 2 back subcutaneous injection 40%CCl4 sweet oil 3ml/kg, totally 6 weeks weekly later on.Except that normal group, the 1st～2 week, each group all gave the Semen Maydis powder feed that 20% lard adds 0.5% SUV at experimental session, and the 3rd～6 week raised with normal diet.Each administration group is irritated the soup that stomach gives corresponding dosage simultaneously when modeling begins, administration volume 1ml/100g, and administration time is totally 12 weeks.Each is cut open the belly under the etherization after organizing the completion of medication cycle, through the lower chamber dooor venous blood collection, detects Serum ALT, AST, Alb respectively, and gets a part of hepatic tissue and process LH, is used to detect oxyproline (Hyp) content.Other gets hepatic tissue and does HE dyeing and be used for pathological study.Experimental result is seen table 6:
The influence of Hyp in the table 6 pair Liver Fibrosis Model liver tissues of rats (X ± S)
Annotate: compare with the blank group,
*P＜0.01;
Experiment 4
Influence to the mouse pulmonary fibrosis model
Experimental technique:
Age in animal: 8-1 week, male, Kunming mouse, body weight 18-22g.Reagent: bleomycin for inj A5, Hebei, Tianjin pharmaceutical factory, prednisone acetate tablets: fairy house pharmaceutical Co. Ltd produces, and the time spent grinding powder is made into 50% suspension with dissolved in distilled water.PBS liquid, oneself is prepared.Adopt bleomycin A5 to duplicate animal diffuse interstitial pulmonary fibrosis model.Mouse is lain on the back on experiment table after with etherization, and fixing head and four limbs cut skin of neck, by the disposable injection bleomycin A5 of tracheae solution 0.05ml (pastille 0.1mg, 5mg/kg).Skin suture after injection finishes, mouse is upright, rotation make soup uniform distribution in lung as far as possible.The operation of surgical procedure Strict aseptic.The blank group is in kind injected equivalent physiologic saline for substitute bleomycin A5, the clear-headed back of animal ad libitum access.The mouse modeling is divided into 6 groups at random after 24 hours, be respectively blank group, Prednisone acetate group (6.5mg/kg), the application's solid preparation group.The blank group is irritated stomach and is given zero(ppm) water 0.2ml/10g, every day three times; Positive controls is irritated stomach and is given Prednisone acetate, 6.5mg/kg, 0.2ml/10g, every day three times after the modeling; The administration group is irritated the soup that stomach gives each corresponding dosage by 0.2ml/10g after the modeling.
More than each the group, successive administration 28 days.The result sees table 7 for details.
The influence of table 7 pair pulmonary fibrosis model mouse lung coefficient (X ± S)
Annotate: compare with the blank group,
*P＜0.01
Experiment 5
The MTT reduction method detects the preparation anti-tumor activity test
Experiment material:
MTT: with phosphate buffered saline buffer (PBS) the dissolving MTT final concentration 5mg/ml of 0.01mol/L, filtration sterilization, 4 ℃ keep in Dark Place after the packing.
The preparation of MTT lysate: the 80g sodium laurylsulfonate is dissolved in the N-N-dimethyl methyl phthalein amine of 200ml, and the heating in water bath hydrotropy adds 200ml zero(ppm) water, mixes with 1N hydrochloric acid (1: 1) with 80% acetate and transfers pH to 4.7.
Cell strain is selected for use: human normal cell line strain, human hepatoma cell strain and National People's Congress's sclc cell line.
Experimental technique: single cell suspension is inoculated in 96 orifice plates (with the RPM-1640 basic medium with cell dilution to 30000/ml, every hole adds the good cell of 200 μ l dilution), cultivates 24 hours under 37 ℃, 5% carbonic acid gas saturated humidity; Every group four parallel appearance; Remove substratum, get new preparing culture medium and prepare cancer therapy drug (the application's preparation) solution by series concentration, every hole 20 μ l cultivated 48 hours; Every hole adds the MTT20 μ l of 2mg/ml, hatches 4 hours; Nutrient solution in the sucking-off hole (as far as possible fully) adds DMSO liquid (150 μ l/ hole), vibrates 10 minutes, and crystallisate is fully dissolved; ELIASA detects each hole OD value, (detecting wavelength 560nm); Draw the cell viability graphic representation, obtain the IC50 value.Experimental result is as shown in table 8:
Table 8 cytotoxicity experiment result
Brief summary: the application's solid preparation has stronger cell toxicant, and its toxicity has certain selectivity to normal cell and cancer cells, and the application's solid preparation can be used for making each antitumor drug.
Experiment 6
Comparison to the rats'liver tumor suppression
Laboratory animal: rat, 150g-180g, male and female are regardless of.
Experiment medicine: saline water; The present invention respectively organizes preparation; Commercially available sodium cantharidinate tablet.
Experimental technique: get rat and be divided into saline water group, sodium cantharidinate tablet group, preparation group of the present invention, make W
256Liver in inoculation, inoculate after 7 days, press the dosage intraperitoneal injection of anesthesia of 35mg/kg with vetanarcol, fixing, cutting open the belly exposes liver, tumor surface maximum diameter (a) and path (b) are pressed (a*b on the measurement liver
2)/2=V (gross tumor volume).Separate stomach, arteria duodenalis, arteria hepatica communis and proper hepatic artery, ligation stomach, arteria duodenalis far-end are with silver brain clip blocking-up arteria hepatica communis; In sending into proper hepatic artery again at the gastroduodenal artery upper cut and after inserting external diameter 0.3mm conduit under the operating loupe, divide into groups to inject respectively to receive reagent thing, postoperative tube drawing ligation gastroduodenal artery by experiment then; Decontrol the arteria hepatica communis silver brain clip, sew up the incision again, place animal housing to wait to revive rat; Continue breeding observing; Performed the operation back 8 days, and detected gross tumor volume by last method, experimental result is seen table 9:
Table 9 is respectively organized preparation to the rejection ratio of tumour
Annotate: compare with the saline water group
*P＜0.01;
Experiment 7
Anti-ageing experiment
Experimental technique:
With aged Shanghai is the mouse random packet, 14/group, and male and female half and half, the administration group is irritated stomach preparation 15mg/kg of the present invention, 3 weeks of administration altogether every day.Blood 50 μ l are got in the mouse docking, measure the activity of SOD according to the autoxidizable method of pyrogallol.With the mouse sacrificed by decapitation, take out liver, inhale with filter paper and remove residual blood, shred and weigh, add saline water, be prepared into 1% homogenate, adopt the thiobarbituricacid method to measure the content of LPO in the hepatic tissue.Experimental result is seen table 10:
Table 10 solid preparation is to the influence of SOD, LPO
Annotate: compare with control group
*P＜0.01,
*P＜0.05; Compare #P＜0.05 with positive controls
Five. preparation embodiment
Embodiment 1
Method 1: the red sage root obtains aqueous extract with water extraction, transfers pH value to 3.5,110 ℃ of temperature, heats 1 hour, and solution filters, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 2: the red sage root obtains aqueous extract with water extraction, transfers pH value to 3.5, puts into the microwave extraction device, and frequency is that 915MHz, power are 1000 watts microwave extraction 0.5 hour, extracting liquid filtering, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 3: the red sage root obtains aqueous extract with water extraction, transfers pH value to 3.5,110 ℃ of temperature, heats 1 hour; Solution filters, and filtrating is separated through the HPD-100 macroporous resin column chromatography, first water, 10% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 30% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Method 4: the red sage root obtains aqueous extract with water extraction, transfers pH value to 3.5, puts into the microwave extraction device, and frequency is that 915MHz, power are 1000 watts microwave extraction 0.5 hour; Extracting liquid filtering, filtrating is separated through the HPD-100A macroporous resin column chromatography, first water, 10% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 30% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Formulation preparation: get above-mentioned danshen root salvianolic acid A, learn the conventional preparation that requires to be prepared into according to tablet, capsule, granule, soft capsule, pellet, pill, oral liquid medicine.
Embodiment 2
Method 1: the red sage root extracts with 70% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5,120 ℃ of temperature, heats 3 hours, and solution filters, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 2: the red sage root extracts with 70% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5; Put into the microwave extraction device, frequency is that 2450MHz, power are 10000 watts microwave extraction 1 hour, extracting liquid filtering; The filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 3: the red sage root extracts with 70% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5,120 ℃ of temperature, heats 3 hours; Solution filters, and filtrating is separated through the HPD-300 macroporous resin column chromatography, first water, 20% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 50% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Method 4: the red sage root extracts with 70% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5, puts into the microwave extraction device; Frequency is that 2450MHz, power are 10000 watts microwave extraction 1 hour, extracting liquid filtering, and filtrating is separated through the HPD-400 macroporous resin column chromatography, first water, 20% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 50% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Formulation preparation: get above-mentioned danshen root salvianolic acid A, learn the conventional preparation that requires to be prepared into according to tablet, capsule, granule, soft capsule, pellet, pill, oral liquid medicine.
Embodiment 3
Method 1: the red sage root extracts with 80% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 6.0,130 ℃ of temperature, heats 6 hours, and solution filters, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 2: the red sage root extracts with 80% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 6.0; Put into the microwave extraction device, frequency is that 2450MHz, power are 15000 watts microwave extraction 2 hours, extracting liquid filtering; The filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 3: the red sage root extracts with 80% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 6.0,130 ℃ of temperature, heats 6 hours; Solution filters, and filtrating is separated through the HPD-400A macroporous resin column chromatography, first water, 30% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 70% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Method 4: the red sage root extracts with 80% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 6.0, puts into the microwave extraction device; Frequency is that 2450MHz, power are 15000 watts microwave extraction 2 hours, extracting liquid filtering, and filtrating is separated through the HPD-450 macroporous resin column chromatography, first water, 30% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 70% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Formulation preparation: get above-mentioned danshen root salvianolic acid A, learn the conventional preparation that requires to be prepared into according to tablet, capsule, granule, soft capsule, pellet, pill, oral liquid medicine.
Embodiment 4
Method 1: the red sage root obtains aqueous extract with water extraction, transfers pH value to 4.0,115 ℃ of temperature, heats 2 hours, and solution filters, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 2: the red sage root extracts with 20% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.0; Put into the microwave extraction device, frequency is that 915MHz, power are 2000 watts microwave extraction 0.8 hour, extracting liquid filtering; The filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 3: the red sage root extracts with 25 ethanolic solns and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers PH value to 4.0,115 ℃ of temperature, heats 1.5 hours; Solution filters, and filtrating is separated through the D101 macroporous resin column chromatography, first water, 15% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 35% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Method 4: the red sage root obtains aqueous extract with water extraction, transfers pH value to 4.0, puts into the microwave extraction device, and frequency is that 915MHz, power are 2500 watts microwave extraction 0.8 hour; Extracting liquid filtering, filtrating is separated through the 1300-I macroporous resin column chromatography, first water, 15% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 35% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Formulation preparation: get above-mentioned danshen root salvianolic acid A, learn the conventional preparation that requires to be prepared into according to tablet, capsule, granule, soft capsule, pellet, pill, oral liquid medicine.
Embodiment 5
Method 1: the red sage root extracts with 75% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 5.5,125 ℃ of temperature, heats 5.5 hours, and solution filters, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 2: red sage root water obtains aqueous extract, transfers pH value to 5.0, puts into the microwave extraction device, and frequency is that 2450MHz, power are 14500 watts microwave extraction 1.8 hours, extracting liquid filtering, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 3: the red sage root obtains aqueous extract with water extraction, transfers pH value to 5.5,125 ℃ of temperature, heats 5 hours; Solution filters, and filtrating is separated through the AB-8 macroporous resin column chromatography, first water, 25% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 65% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Method 4: the red sage root extracts with 75% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 5.5, puts into the microwave extraction device; Frequency is that 2450MHz, power are 14500 watts microwave extraction 1.8 hours, extracting liquid filtering, and filtrating is separated through 1400 macroporous resin column chromatographies, first water, 25% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 65% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Formulation preparation: get above-mentioned danshen root salvianolic acid A, learn the conventional preparation that requires to be prepared into according to tablet, capsule, granule, soft capsule, pellet, pill, oral liquid medicine.
Embodiment 6
Method 1: the red sage root obtains aqueous extract with water extraction, transfers pH value to 4.5,120 ℃ of temperature, heats 4.5 hours, and solution filters, and the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 2: the red sage root extracts with 70% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5; Put into the microwave extraction device, frequency is that 2450MHz, power are 10000 watts microwave extraction 1 hour, extracting liquid filtering; The filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 3: the red sage root extracts with 70% ethanolic soln and obtains aqueous extract or alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5,120 ℃ of temperature, heats 3 hours; Solution filters, and filtrating is separated through 1400 macroporous resin column chromatographies, first water, 20% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 50% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Method 4: the red sage root extracts with 70% ethanolic soln and obtains alcohol extract, and alcohol extract concentrates ethanol to the greatest extent, transfers pH value to 4.5, puts into the microwave extraction device; Frequency is that 2450MHz, power are 9000 watts microwave extraction 1.5 hours, extracting liquid filtering, and filtrating 1300-I macroporous resin column chromatography separates first water, 20% Diluted Alcohol wash-out; Remove impurity, use the ethanol elution of 45% concentration again, collect elutriant; Reclaim ethanol to most, drying obtains the danshen root salvianolic acid A extract;
Formulation preparation: get above-mentioned danshen root salvianolic acid A, learn the conventional preparation that requires to be prepared into according to tablet, capsule, granule, soft capsule, pellet, pill, oral liquid medicine.
Claims (2)
1. the preparation method of a danshen root salvianolic acid A is characterized in that:
Danshen root salvianolic acid A extracts:
Method 1: red sage root water or ethanolic soln extract and obtain aqueous extract or alcohol extract; Alcohol extract concentrates ethanol to most, and adjust pH is put into the microwave extraction device to 3.5-6.0; Frequency is that 915MHz-2450MHz, power are 1000-15000 watt microwave extraction 0.5-2 hour; Extracting liquid filtering, the filtrating concentrate drying obtains the danshen root salvianolic acid A extract;
Method 2: red sage root water or ethanolic soln extract and obtain aqueous extract or alcohol extract, and alcohol extract concentrates ethanol to most, and adjust pH is to 3.5-6.0; Put into the microwave extraction device, frequency is that 915MHz-2450MHz, power are 1000-15000 watt microwave extraction 0.5-2 hour, extracting liquid filtering; Filtrating is separated through nonpolar or low-pole macroporous resin column chromatography, and first water, 10-30% Diluted Alcohol wash-out are removed impurity; Use the ethanol elution of 30-70% concentration again, collect elutriant, reclaim ethanol to most; Drying obtains the danshen root salvianolic acid A extract.
2. the preparation method of a kind of danshen root salvianolic acid A according to claim 1, wherein said macroporous resin column are HPD-100, HPD-100A, HPD-300, HPD-400, HPD-400A, HPD-450, D101,1300-I, 1400 or AB-8.
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| CN1397276A (en) * | 2002-08-07 | 2003-02-19 | 沈阳药科大学 | Preparing process and medical application of SLA-A contained in red sage root and its composition |
| CN1830947A (en) * | 2006-04-21 | 2006-09-13 | 王国振 | Method for extracting 'Danfen' phenolic acid-A |
| CN1887849A (en) * | 2006-07-13 | 2007-01-03 | 正大青春宝药业有限公司 | Salvianolic acid A preparing process |
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| CN1397276A (en) * | 2002-08-07 | 2003-02-19 | 沈阳药科大学 | Preparing process and medical application of SLA-A contained in red sage root and its composition |
| CN1830947A (en) * | 2006-04-21 | 2006-09-13 | 王国振 | Method for extracting 'Danfen' phenolic acid-A |
| CN1887849A (en) * | 2006-07-13 | 2007-01-03 | 正大青春宝药业有限公司 | Salvianolic acid A preparing process |
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