CN106431915A - Method for preparing salvianolic acid A - Google Patents
Method for preparing salvianolic acid A Download PDFInfo
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- CN106431915A CN106431915A CN201610806533.8A CN201610806533A CN106431915A CN 106431915 A CN106431915 A CN 106431915A CN 201610806533 A CN201610806533 A CN 201610806533A CN 106431915 A CN106431915 A CN 106431915A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/30—Preparation of carboxylic acid esters by modifying the acid moiety of the ester, such modification not being an introduction of an ester group
- C07C67/333—Preparation of carboxylic acid esters by modifying the acid moiety of the ester, such modification not being an introduction of an ester group by isomerisation; by change of size of the carbon skeleton
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
- C07C67/58—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
Abstract
The invention discloses a method for preparing salvianolic acid A. The method includes the following steps that firstly, salvianolic acid B is prepared into a solution with the concentration of 35-45 mg/mL by means of NaOH or NaHCO3 with the pH value of 3.5-4.5, the solution is placed in a subcritical water reaction kettle, after the temperature of a heating furnace reaches 170-190 DEG C and is stabilized, the reaction kettle is placed into the heating furnace, the reaction kettle is taken out after 50-70 min and placed in ice water bath or cold water to be cooled, the liquid is taken out and subjected to freeze-drying, and a crude product rich in salvianolic acid A is obtained; secondly, salvianolic acid A is separated and purified by means of high-speed countercurrent chromatography, wherein a solvent system is prepared from petroleum ether, ethyl acetate, n-butyl alcohol and water according to the ratio of 2:3:1:9, 10 mM of trifluoroacetic acid is added to an upper phase to form a stationary phase, 10 mM ammonia water is a lower phase and serves as a mobile phase, the volume of a high-speed countercurrent chromatography column is 200-400 mL, the sample loading amount is 1.0-1.2 g, the rotation speed is 600-1000 rpm, the flow speed is 1-4 mL/min, and the detection wavelength is 280 nm. The method is low in cost, easy to operate and high in efficiency, salvianolic acid crude extracts can be converted on a large scale, and a salvianolic acid A monomeric compound with the purity higher than 98% is separated and prepared.
Description
Technical field
The invention belongs to the efficient preparing technical field of Effective Component of Chinese Medicine, and in particular to a kind of preparation side of salvianolic acid A
Method.
Background technology
Radix Salviae Miltiorrhizae is dry root and the rhizome of labiate Radix Salviae Miltiorrhizae Salvia miltiorrhizaBge., dispels with promoting blood circulation
The stasis of blood, inducing menstruation to relieve menalgia, relieving restlessness that clears away heart-fire, the effects such as cool blood to disappear carbuncle.For obstruction of qi in the chest and cardialgia, gastral cavity abdomen hypochondriac pain, abdominal mass is gathered, hot numbness pain,
Dysphoria and insomnia, menoxenia, dysmenorrhea amenorrhea, the disease such as sore swell and ache curative.In the last few years, research found that Radix Salviae Miltiorrhizae had coronary artery dilating, resisted
Platelet aggregation, improves blood circulation, prevents thrombosiss and atheromatous plaque from being formed, promote pathological change to recover etc.
Effect, and it is widely used in the ischemic diseasess such as quality cardiovascular and cerebrovascular vessel.Radix Salviae Miltiorrhizae has become China's consumption maximum, sale at present
The most Chinese medicine of volume highest, preparation manufacturer, the red sage formulation of Clinical practice mainly includes：FUFANG DANSHEN DIWAN, compound recipe pellet
Ginseng injection, FUFANG DANSHEN PIAN, poly phenolic acid of Radix Salviae Miltiorrhizae salt injection etc..In Radix Salviae Miltiorrhizae, chemical composition is broadly divided into two big class：Water solublity
Composition, i.e. phenolic acid compound；Liposoluble constituent, i.e. diterpene quinone.The end of the sixties, research worker was to Salvia miltiorrhiza Bge water so far
Soluble components have made intensive studies, it was demonstrated that its principle active component is phenolic acid compound.The salvia-soluble that reports earliest
Composition is protocatechualdehyde, and report danshensu was the basic structure of various salvianolic acid compounds later.Isolated afterwards a series of
Pressure differential self, such as salvianolic acid A～K, rosmarinic acid, alkannic acid etc., they all have anti peroxidation of lipid and remove certainly
Acted on by base, wherein salvianolic acid A (Salvianolic acid A) activity is most strong.
The existing preparation method with regard to salvianolic acid A has direct preparation method and indirect reformer method.Directly preparation method is divided into two kinds,
A kind of is direct synthesis technique (as patent CN201410216134.7), using salvianolic acid A synthesis precursor through demethylating reaction system
Standby salvianolic acid A, then through preparation HPLC purification, purity can bring up to 97%；A kind of is direct method of isolation, (as patent
CN201010541651.3), with red rooted salvia as raw material, the method such as extracted, rough segmentation, decolouring, purification, obtain high-purity pellet phenol
Sour A monomer.Indirect reformer method (as patent CN201310487751.6) is will be heated to Radix Salviae Miltiorrhizae or salvianolic acid B, and acid adjustment alkali is anti-
Some hours are answered, and salvianolic acid B is converted into salvianolic acid A, then high-purity danshinolic acid A is obtained through preparative separation.Directly prepare method and
Indirect reformer method all has the shortcomings of preparation efficiency is low, and conversion rate is slow, while the component for obtaining rich in salvianolic acid A needs to combine
Polyamide chromatograph, macroporous resin, silica gel column chromatography and preparative liquid chromatography carry out separating, and waste time and energy, pollute environment, sample
Purity is low, and column chromatography has irreversibility adsorption to sample repeatedly, isolated salvianolic acid A monomer preparation efficiency is low,
Relatively costly, it is difficult to develop into the big isolation technics of preparation amount.
More than boiling point is heated water to, below critical point, and control system pressure makes water that liquid is remained, this state
Water is referred to as subcritical water.Under usual conditions, water is polar compound, under 505kPa pressure, raises (50～300 with temperature
DEG C), its dielectric constant is decreased to 1 by 70, that is to say, that its property by highly polar fade to nonpolar, can by solute by polarity by
High to Low extract.Under conditions of temperature and pressure is all higher, the polarity reduction of water, can be with extracting apolar compound；
Under conditions of temperature and pressure is all relatively low, the polarity of water is improved, and can extract polar compound.Due to be do not use acid, alkali and
Treatment technology of the water of catalyst under high thermal high, therefore the extracting method of subcritical water be referred to as " green process
Method ".Additionally, extract can complete within the short time of several seconds to several minutes, have the advantages that continuous processing can be carried out.With
When subcritical water there is " strong dissolved organic matter is in water " and " strong decomposing force " be equal to the different property of light water.Profit
This property is used, subcritical water is utilized to extract useful component (including extracting the analyte for producing with decomposition reaction).Sub-
Critical technology for hydrolyzing is the focus in natural product extraction and Study on Transformation field in recent years.Which is used for natural product extraction conversion tool
Have the advantages that fast, the solvent-free pollution of reaction rate, early stage and post-processing step be simple, high income, energy consumption are low unique.
High speed adverse current chromatogram (High-speed Counter-current Chromatography, HSCCC) is nearly 30 years
A kind of efficient, the quick liquid liquid partition chromatography isolation technics continuously without the need for any solid support for growing up, it keeps away
Sample that solid state adhesion body or carrier bring is exempted from easily by various problems such as dead absorption, loss and degeneration.The refined adverse current of pH- zone
Chromatograph (pH-Zone-Refining Countercurrent Chromatography) be then in common high speed adverse current chromatogram
On the basis of instrument, by the allotment of the composition to separating the solvent system used by sample, using the means of chemistry, sample sets are made
The chromatographic separation process for dividing adds the feature by the aggregation of pH zone, meanwhile, make the elution process of component show as being similar to displacement
The elution process of (replacement) chromatograph (Displacement Chromatography), therefore, its chromatogram is no longer that Gauss divides
The chromatographic peak profile series of cloth, and the precipitous rectangle zone series in the border that becomes the big minispread by pH value, its result is can handle
The separation preparation amount of the adverse current chromatogram instrument of same volume improves several times or even ten times.
Content of the invention
It is an object of the invention to provide a kind of preparation method of salvianolic acid A.
A kind of preparation method of salvianolic acid A, step is as follows：
(1) by the salvianolic acid B NaOH or NaHCO that pH is 3.5-4.5 (preferably 4.0)3(NaHCO3Advantage：Alkalescence is weaker,
Reaction is gentleer) water is configured to 35-45mg/mL (preferably：Solution 40mg/mL), is placed in subcritical water stainless steel cauldron,
Heating furnace reaches 170 DEG C -190 DEG C (preferably：180 DEG C) and stable after, reactor is put into heating furnace, 50min-70min is (preferably
60min) take out rapidly reactor afterwards and be put into ice-water bath or cold water punching (advantage：Fast cooling, prevents from continuing during cooling
Continuous reaction) middle cooling, liquid is taken out, lyophilization, obtain rich in salvianolic acid A crude product；
(2) application high speed adverse current chromatogram isolates and purifies salvianolic acid A：Solvent system is petroleum ether：Ethyl acetate：N-butyl alcohol：
Water=2:3:1:9, upper addition 10mM trifluoroacetic acid be fixing phase, lower mutually for 10mM ammonia be mobile phase, high-speed counter-current chromatograph
Column volume is 200-400 (preferably 300) mL, applied sample amount 1.0-1.2g (preferably 1.2g), and rotating speed 600-1000rpm is (preferably：
800rpm), upper is mutually fixing phase, and lower is mutually that mobile phase, flow velocity 1-4mL/min is (preferably：2.0mL/min), fixing phase retention rate
57%, Detection wavelength 280nm.
A kind of have coronary artery dilating, antiplatelet aggregation, improve blood circulation, prevent thrombosiss, atherosclerosiss
Speckle forms, promotes the preparation method of the medicine of pathological change recovery, the step of with above-mentioned salvianolic acid A preparation method.
Beneficial effects of the present invention：
1st, salvianolic acid B is converted into salvianolic acid A crude product using supercritical water conversion by the preparation method of the present invention, is eventually passed
High speed adverse current chromatogram isolates and purifies salvianolic acid A, and cost is less than prior art, easy to operate, efficiency high, can relatively high-volume by pellet
Phenolic acid crude extract is converted, and salvianolic acid A monomeric compound of the purity more than 98% is prepared in separation.
2nd, the method having at present using extracting fat soluble component in Chinese medicine red sage root by subcritical water etc., but these methods are all
Make use of subcritical water under conditions of temperature and pressure is all higher, the polarity of water reduces, the effect of extracting apolar compound,
And the present invention makes full use of subcritical water to Tanshin Water-soluble Ingredient dissolubility height, the extraction of Related Component is cannot be only used for,
While also using the feature of High Temperature High Pressure, salvianolic acid B is converted into the higher salvianolic acid A of biological activity.
3rd, by the salvianolic acid B NaHCO that pH is 4.03Water is configured to the solution purpose of 40mg/mL：The experiment proved that, in pellet
Add acid or alkali in phenolic acid B, can all accelerate the conversion rate of salvianolic acid B, but only when pH is 4.0 or so, most have
Convert to the direction of salvianolic acid A beneficial to salvianolic acid B, make the yield highest of salvianolic acid A.Advantage：The product of salvianolic acid A can both have been improved
Amount, can accelerate the conversion rate of salvianolic acid B again.
4th, after heating furnace reaches 180 DEG C and stablizes, reactor is put into heating furnace, the purpose after 60min：Heating furnace reacts
Before reach steady statue, be conducive to carrying out control on time and temperature to entirely reacting, be improved the repeatability of reaction.
Advantage：Efficient energy-saving, conversion are complete.
Description of the drawings
Fig. 1 is the process chart of embodiment 1；
Fig. 2 is that embodiment 1 is rich in the detached zone adverse current chromatogram figure of salvianolic acid A crude product, a：Danshensu；b：Salvianolic acid D；c：
Salvianolic acid A；d：Protocatechualdehyde；
Fig. 3 is the high-efficient liquid phase chromatogram of 1 salvianolic acid B sample of embodiment；
Fig. 4 is high-efficient liquid phase chromatogram of the embodiment 1 rich in salvianolic acid A crude product；
Fig. 5 is the high-efficient liquid phase chromatogram of 1 danshensu monomer of embodiment；
Fig. 6 is the high-efficient liquid phase chromatogram of 1 salvianolic acid D monomer of embodiment；
Fig. 7 is the high-efficient liquid phase chromatogram of 1 salvianolic acid A monomer of embodiment；
Fig. 8 is the high-efficient liquid phase chromatogram of 1 protocatechualdehyde monomer of embodiment；
Fig. 9 is the zone adverse current chromatogram figure of comparative example 1；
Figure 10 is the zone adverse current chromatogram figure of comparative example 2.
Specific embodiment
The invention will be further described with embodiment below in conjunction with the accompanying drawings.
Embodiment 1
As shown in Fig. 1 salvianolic acid A monomer preparation flow figure.
By salvianolic acid B (Fig. 3) NaHCO that pH is 4.03Water is configured to the salvianolic acid B solution of 40mg/mL, takes on 40mL
State solution to be placed in 50mL subcritical water stainless steel cauldron, after heating furnace reaches 180 DEG C and stablizes, reactor is put into heating
Stove, starts timing.Reactor being taken out rapidly after 60min and is put in ice-water bath and cool down, liquid is taken out, lyophilization, obtain rich
Crude product containing salvianolic acid A (Fig. 4).
2. application high speed adverse current chromatogram isolates and purifies salvianolic acid A
Solvent system is petroleum ether：Ethyl acetate：N-butyl alcohol：Water=2:3:1:9, upper addition 10Mm trifluoroacetic acid is for fixing
Phase, lower mutually for 10mM ammonia be mobile phase, high-speed counter-current chromatograph column volume be 300mL, applied sample amount 1.2g, rotating speed 800rpm,
Upper is mutually fixing phase, and lower is mutually mobile phase, flow velocity 2.0mL/min, fixing phase retention rate 57%, Detection wavelength 280nm.
Specifically operating procedure is：Solvent system is prepared by above-mentioned solvent ratios, be placed in separatory funnel, stand after shaking up
Layering, ready to balance for a period of time afterwards will be biphase separate up and down, and upper addition 10mM trifluoroacetic acid is fixing phase, and lower is mutually 10Mm ammonia
For mobile phase, 1.2g is taken rich in salvianolic acid A crude product, be dissolved in 5mL and add the upper phase of 10mM trifluoroacetic acid and 5mL to be not added with ammonia
Stand-by in phase.The semi-preparative high-speed counter-current chromatograph that is developed with field company using Shanghai, it be by plunger displacement pump, injection valve, purple
Outer detector, monitor and chromatography column (spiral tube for being formed by polyfluortetraethylene pipe multi-lay winding, capacity is 300mL)
Deng composition, make injection valve in sample introduction state first, fixing phase pump is filled chromatography column with certain flow rate, termination of pumping.Open
Speed control is opened, is rotated forward the chromatographic chromatography column of high velocity stream, during turn up 800rpm, the sample for having dissolved is noted
In the liquid storage tube of emitter injection counter-current chromatograph injection valve, rotation injection valve makes sample enter chromatography column for connecing column state.
Setting flow rate of mobile phase is 2.0mL/min, starts pump mobile phase, then receives target according to detector ultraviolet spectrogram (Fig. 2)
Composition, obtains danshensu (38.9mg, Fig. 5), salvianolic acid D (9.5mg, Fig. 6), salvianolic acid A (227.3mg, Fig. 7) and protocatechualdehyde
(32.8mg, Fig. 8), HPLC purity assay is more than 98%.
Using efficient liquid phase chromatographic analysis separator, liquid-phase condition：Kromasil 100-5C18Post (4.6 × 250mm) is purple
Outer Detection wavelength 286nm, column temperature：25 DEG C, flow velocity：1.0mL/min, sample size：10 μ L, mobile phase adopts acetonitrile (A) and 0.2%
Aqueous formic acid (B) gradient elution, gradient condition is as follows：0-9min, 10%-22%A；9-19min, 22%-24%A；19-
35min, 24%A；35-43min, 24%-36%A；43-48min, 36%-100%A；48-50min, 100%A.
Structural Identification：To isolated alkaloid application Agilent 5973N mass spectrograph and Varian 600MHz nuclear-magnetism
Resonance spectrometer carries out MS respectively,1The measure of HNMR spectrum, the data obtained is as follows：
Danshensu：ESI-MS,m/z,197[M-H]-,395[2M-H]-,135[M-H-H2O-CO2]-.1H-NMR(DMSO-
d6,400MHz)δ:2.51 (1H, dd, J=5.6,9.2Hz, H-7), 2.85 (1H, dd, J=2.4,9.2Hz, H-7), 3.90
(1H, dd, J=2.5,5.6Hz, H-8), 6.45 (1H, dd, J=1.3,5.3Hz, H-6), 6.61 (1H, d, J=5.3Hz, H-
5),6.65(1H,s,H-2).13C-NMR(DMSO-d6,100MHz)δ:49.1(C-7),72.6(C-8),115.7(C-5),
117.5(C-2),120.5(C-6),130.5(C-1),143.9(C-3),145.2(C-4),177.1(C-9).
Salvianolic acid D：ESI-MS,m/z,417[M-H]-,373[M-H-CO2]-,197[C9H10O5-H]-,175[M-H-CO2-
C9H10O5]-.1H-NMR(DMSO-d6,400MHz)δ:2.96 (2H, m, H-7 "), 4.85 (1H, dd, J=8.1,8.5Hz, H-
8″),3.58(2H,s,Ar-CH2), 6.24 (1H, d, J=16.0Hz, H-8), 7.86 (1H, d, J=16.0, H-7), 6.47-
7.07(5H,Ar-H).13C-NMR(DMSO-d6,100MHz)δ:37.5(C-2′),49.1(C-7″),75.0(C-8″),114.1
(C-5),115.8(C-3″),116.1(C-5″),117.4(C-8),118.6(C-6),119.9(C-2),124.44(C-6″),
125.8(C-1),128.9(C-1″),143.3(C-4″),144.0(C-7),144.4(C-4),145,7(C-3″),149.1(C-
3),166.3(C-9),172.0(C-9″),175.5(C-1′).
Salvianolic acid A：ESI-MS,m/z,494[M-H]-,295[M-H-C9H10O5]-.1H-NMR(DMSO-d6,400MHz)δ:
2.75 (1H, dd, J=14.3,8.5Hz, H-7 '), 2.98 (1H, dd, J=14.4/4.5Hz, H-7 '), 4.90 (1H, dd, J=
8.5,6.0Hz, H-8 '), 6.26 (1H, d, J=16.0Hz, H-8), 6.45 (1H, dd, J=8.3,2.0Hz, H-6 "), 6.54
(1H, d, J=8.5Hz, H-5 '), 6.53 (1H, d, J=16.0Hz, H-7 "), 6.63 (1H, d, J=2.0Hz, H-2 '), 6.72
(1H, d, J=8.7Hz, H-5), 7.12 (1H, d, J=8.6Hz, H-6), 6.77 (1H, d, J=8.0Hz, H-5 "), 6.86 (1H,
Dd, J=8.3,2.0Hz, H-6 "), 7.04 (1H, d, J=2.0Hz, H-2 "), 7.13 (1H, d, J=16.0Hz, H-8 ").13C-
NMR(DMSO-d6,100MHz)δ:36.8(C-7′),75.0(C-8′),112.8(C-2″),114.3(C-5),115.3(C-8),
115.7(C-5′),116.4(C-5″),118.6(C-2′),119.0(C-8″),119.1(C-6),119.8(C-6″),123.7
(C-6′),126.5(C-1),129.0(C-2),134.2(C-1′),135.3(C-3),143.6(C-7″),144.8(C-1″),
145.6(C-4′),145.6(C-3′),145.6(C-3″),145.7(C-4″),147.1(C-7),148.4(C-4),166.2
(C-9),171.8(C-9′)..
Protocatechualdehyde：ESI-MS,m/z,137[M-H]-,109[M-H-CO]-.1H-NMR(DMSO-d6,400MHz)δ:
9.69 (1H, s, H-7), 7.27 (1H, dd, J=1.3/5.4Hz, H-6), 7.23 (1H, d, J=1.2Hz, H-2), 6.89 (1H,
D, J=5.4Hz, H-5).13C-NMR(DMSO-d6,100MHz)δ:114.7(C-5),115.9(C-2),125.1(C-6),
129.1(C-1),146.4(C-3),153.0(C-4),191.3(-CHO).
Embodiment 2
By the salvianolic acid B NaHCO that pH is 3.53Water is configured to the salvianolic acid B solution of 45mg/mL, takes the above-mentioned solution of 40mL
It is placed in 50mL subcritical water stainless steel cauldron, after heating furnace reaches 182 DEG C and stablizes, reactor is put into heating furnace, is opened
Beginning timing.Reactor being taken out rapidly after 55min and is put in ice-water bath and cool down, liquid is taken out, lyophilization, obtain rich in red phenol
Sour A crude product.
Application high speed adverse current chromatogram isolates and purifies salvianolic acid A
Solvent system is petroleum ether：Ethyl acetate：N-butyl alcohol：Water=2:3:1:9, upper addition 10mM trifluoroacetic acid is for fixing
Phase, lower mutually for 10mM ammonia be mobile phase, high-speed counter-current chromatograph column volume be 300mL, applied sample amount 1.0g, rotating speed 750rpm,
Upper is mutually fixing phase, and lower is mutually mobile phase, flow velocity 2.0mL/min, fixing phase retention rate 55%, Detection wavelength 280nm.
Specifically operating procedure is：Solvent system is prepared by above-mentioned solvent ratios, be placed in separatory funnel, stand after shaking up
Layering, ready to balance for a period of time afterwards will be biphase separate up and down, and upper addition 10mM trifluoroacetic acid is fixing phase, and lower is mutually 10mM ammonia
For mobile phase, 1.2g is taken rich in salvianolic acid A crude product, be dissolved in 5mL and add the upper phase of 10Mm trifluoroacetic acid and 5mL to be not added with ammonia
Stand-by in phase.The semi-preparative high-speed counter-current chromatograph that is developed with field company using Shanghai, it be by plunger displacement pump, injection valve, purple
Outer detector, monitor and chromatography column (spiral tube for being formed by polyfluortetraethylene pipe multi-lay winding, capacity is 300mL)
Deng composition, make injection valve in sample introduction state first, fixing phase pump is filled chromatography column with certain flow rate, termination of pumping.Open
Speed control is opened, is rotated forward the chromatographic chromatography column of high velocity stream, during turn up 800rpm, the sample for having dissolved is noted
In the liquid storage tube of emitter injection counter-current chromatograph injection valve, rotation injection valve makes sample enter chromatography column for connecing column state.
Setting flow rate of mobile phase is 2.0mL/min, starts pump mobile phase, then receives target component according to detector ultraviolet spectrogram,
Danshensu (36.7mg), salvianolic acid D (9.2mg), salvianolic acid A (219.6mg) and protocatechualdehyde (32.1mg) is obtained, HPLC analysis is pure
Degree is more than 98%.
Comparative example 1：
Condition：Petroleum ether-ethyl acetate-methanol-water (2:3:3:9), upper phase 10mM trifluoroacetic acid, lower phase 10mM ammonia,
Flow velocity：2mL min-1, sample size：1000mg, fixing phase retains：52%, rotating speed：800rpm, other conditions with operation all with reality
Apply as in example 1.Separating resulting：Appearance time is too fast, and compound is separated, such as accompanying drawing 9.
Comparative example 2：
Condition：Petroleum ether-ethyl acetate-methanol-water (2:3:1:9), upper phase 10mM trifluoroacetic acid, lower phase 10mM ammonia,
Flow velocity：2mL min-1, sample size：900mg, fixing phase retains：56%, rotating speed：800rpm, other conditions with operation all with enforcement
As in example 1.Separating resulting：Separation condition makes moderate progress, and salvianolic acid A obtains part sterling, but major part is changed with other
Compound is mixed, and is efficiently separated not yet, such as accompanying drawing 10.
Comparative example 1 be can be seen that with embodiment 1 by Fig. 9 and Figure 10 is to have changed n-butyl alcohol into methanol in solvent system,
And the ratio of solvent system there occurs change, result in that appearance time is too fast, compound is separated, and comparative example 2 is only
It is only to have changed n-butyl alcohol into methanol in solvent system, the ratio of solvent system does not all change, and has again resulted in and has not obtained
Efficiently separate.
Although the above-mentioned accompanying drawing that combines is described to the specific embodiment of the present invention, not model is protected to the present invention
The restriction that encloses, one of ordinary skill in the art are should be understood that on the basis of technical scheme, and those skilled in the art are not
The various modifications that makes by needing to pay creative work or deformation are still within protection scope of the present invention.
Claims (9)
1. a kind of preparation method of salvianolic acid A, is characterized in that：Step is as follows：
(1) by the salvianolic acid B NaOH that pH is 3.5-4.5 or NaHCO3Water is configured to the solution of 35-45mg/mL, is placed in subcritical
In water reactor, after heating furnace reaches 170 DEG C -190 DEG C and stablizes, reactor is put into heating furnace, takes out after 50min-70min
Reactor is simultaneously put into ice-water bath or cold water punching cooling, liquid is taken out, lyophilization, obtains rich in salvianolic acid A crude product；
(2) application high speed adverse current chromatogram isolates and purifies salvianolic acid A：Solvent system is petroleum ether：Ethyl acetate：N-butyl alcohol：Water=
2:3:1:9, upper addition 10mM trifluoroacetic acid be fixing phase, lower mutually for 10mM ammonia be mobile phase, high-speed counter-current chromatograph cylinder
Product is 200-400mL, applied sample amount 1.0-1.2g, rotating speed 600-1000rpm, flow velocity 1-4mL/min, Detection wavelength 280nm.
2. preparation method as claimed in claim 1, is characterized in that：In step (1) salvianolic acid B is 4.0 with pH
NaHCO3Water is configured to the solution of 40mg/mL.
3. preparation method as claimed in claim 1, is characterized in that：In step (1), heating furnace reaches 180 DEG C.
4. preparation method as claimed in claim 1, is characterized in that：Reactor is taken out rapidly simultaneously after 60min in step (1)
It is put in ice-water bath and cools down.
5. preparation method as claimed in claim 1, is characterized in that：Step (2) high speed counter-current chromatograph column volume is
300mL.
6. preparation method as claimed in claim 1, is characterized in that：Applied sample amount 1.2g in step (2).
7. preparation method as claimed in claim 1, is characterized in that：Rotating speed 800rpm in step (2).
8. preparation method as claimed in claim 1, is characterized in that：Flow velocity 2.0mL/min in step (2).
9. a kind of have coronary artery dilating, antiplatelet aggregation, improve blood circulation, prevent thrombosiss, atherosclerotic plaque
Block forms, promotes the preparation method of the medicine of pathological change recovery, it is characterized in that：Including the arbitrary described system of claim 1-8
The step of Preparation Method.
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| CN201610806533.8A CN106431915B (en) | 2016-09-07 | 2016-09-07 | A kind of preparation method of salviandic acid A |
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| CN201610806533.8A CN106431915B (en) | 2016-09-07 | 2016-09-07 | A kind of preparation method of salviandic acid A |
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| CN106431915A true CN106431915A (en) | 2017-02-22 |
| CN106431915B CN106431915B (en) | 2019-04-23 |
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| CN201610806533.8A Active CN106431915B (en) | 2016-09-07 | 2016-09-07 | A kind of preparation method of salviandic acid A |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1995027A (en) * | 2006-12-30 | 2007-07-11 | 山东省分析测试中心 | Separating preparation process of salvianolic acid B |
| CN102249920A (en) * | 2011-05-30 | 2011-11-23 | 上海朗萨医药科技有限公司 | Preparation method of high-purity salvianolic acid A |
| CN105085266A (en) * | 2014-05-14 | 2015-11-25 | 南京虹桥医药技术研究所 | Method for preparing salvianolic acid A from a plurality of salvia plants |
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| CN1995027A (en) * | 2006-12-30 | 2007-07-11 | 山东省分析测试中心 | Separating preparation process of salvianolic acid B |
| CN102249920A (en) * | 2011-05-30 | 2011-11-23 | 上海朗萨医药科技有限公司 | Preparation method of high-purity salvianolic acid A |
| CN105085266A (en) * | 2014-05-14 | 2015-11-25 | 南京虹桥医药技术研究所 | Method for preparing salvianolic acid A from a plurality of salvia plants |
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