Isoflavones Lignanoids compounds extracted from camel thorn plants and uses thereof and extracting method
Technical field:
The present invention relates to a kind of isoflavones Lignanoids compounds and uses thereof and extracting method, particularly a kind of isoflavones Lignanoids compounds extracted and uses thereof and extracting method from camel thorn plants.
Background technology:
Isoflavones lignanoid (isoflavonolignan) compounds refers to the compounds that isoflavones parent and C6-C3 (being Ph-C3) fragment a pair of horses going side by side occur and form by the diox structure.In natural product, there is the compound of similar a pair of horses going side by side and mode that flavanolignan's compounds and coumarinolignan's compounds are arranged, and about the relevant report of this two compounds early, and in recent years just occur for the report of isoflavones Lignanoids compounds.
The isoflavones Lignanoids compounds distributes and to only limit to pulse family in vegitabilia, and its structure type just at present bibliographical information also only have two kinds, the compound of the upper lignanoid of parent compound a pair of horses going side by side fragment, but it is narrower to obtain at present the approach of this class material.
When the bioactive ingredients of leguminous plants camel thorn is carried out to isolation identification, isolated a kind of isoflavones Lignanoids compounds of new structure type.Although also few to the research of this compounds at present, with regard to current progress, this compounds has certain meaning on plant chemotaxonomy.Continue chemistry and the pharmacological action of this compound of further investigation, the isoflavones Lignanoids compounds of finding this new texture have as anticancer, protect the liver, antiviral, enzyme suppresses and the important physiologically active such as platelet activation factor antagonism.
Therefore, the purposes of a kind of isoflavones Lignanoids compounds and this compound and extracting method, particularly a kind of isoflavones Lignanoids compounds extracted from camel thorn plants and the purposes of this compound and the proposition of extracting method, for taking full advantage of plant resources, exploitation is efficient, the natural drug of low toxicity just has great importance.
Summary of the invention:
The purposes and the extracting method that the purpose of this invention is to provide a kind of isoflavones Lignanoids compounds and this compound, particularly purposes and the extracting method of a kind of isoflavones Lignanoids compounds extracted from camel thorn plants and this compound.
The structural formula of isoflavones Lignanoids compounds of the present invention is:
Its expression formula is:
2,3-trans-8-(3-hydroxy-4-methoxyphenyl)-3-(4-hydroxy-3-methoxyphenylmethanol)-2,3-dihydro-7H-1,4-dioxino[2,3-h]chrome-7-one。
Through experimental verification, above-claimed cpd is all inhibited to PDE4, PDE5, and Papaverine is strong than PDE4 to the PDE5 restraining effect, therefore, compound of the present invention all has active effect for treating the difficulty of erecing, increase immunity and antisepsis and anti-inflammation, but applied research is in treating the erection difficulty, increasing in immunity and antisepsis and anti-inflammation one class medicine.
The extracting method of above-claimed cpd is as follows:
Raw material is prepared: get the dry former plant of camel thorn pulverize or be cut into≤particle of 2 centimetres or section be as raw material;
Alcohol extracting: add the ratio refluxing extraction 1~2h of 10kg raw material in every 2500~3500ml, 50~70% ethanol, cross 38~180 μ m aperture sieve, get filtered liquid.Above-mentioned extraction preferably repeats 2~5 times.
Merging filtrate, be concentrated into alcohol concn≤2% and obtain concentrated solution;
Extraction: concentrated solution is extracted to 1~2h in the chloroform of concentrated solution volume 2-3 times, cross 38~180 μ m sieves;
Above-mentioned extraction preferably repeats 2-5 time,
The gained extraction liquid is concentrated into to chloroform concentration≤2%,
Silica gel column chromatography: by 8~12 times of silicagel column cylinder volumes, measure chloroform extraction liquid through silica gel column chromatography;
Described silicagel column particle diameter is preferably below 150 μ m, is preferably 74~150 μ m.
Wash-out: the sherwood oil that is 10~50% with acetone content-acetone mixed solution wash-out
Collect elutriant, process ODS high performance liquid phase preparative column, slough sherwood oil by gained moving phase and acetone obtain isoflavones Lignanoids compounds of the present invention.
Above-mentioned ODS high performance liquid phase preparative column is preferably anti-phase ODS high performance liquid phase preparative column.
Below the granularity requirements 61 μ m of the above-mentioned stationary phase of above-mentioned high performance liquid phase preparative column, preferably below 45 μ m.
Isoflavones Lignanoids compounds of the present invention is analyzed, and result is as follows:
1H(300MHz),
13C(600MHz),and?HMBC?NMR?data?of?compound(C
5H
5N,δppm)
Evidence to isoflavones Lignanoids compounds of the present invention:
Isoflavones lignin compound of the present invention is specific PDE5, PDE4 inhibitor, it can be developed into to oral preparations to be used for the treatment of the difficulty of erecing, to increase immunity and antibacterial and anti-inflammation functions.
Pharmacological testing is as follows:
PDE5 separates from human body platelet, PDE4 is that containing Uniform B uffer solution, [the 2mmolHEPES damping fluid comprises 0.25mol/L sucrose from the freezing blood sample of the every 1ml of rat liver tissue extraction, 1mmol/L EDTA, 1mmol/L (phenyl-methylsulfonyl-fluorochemical) PMSF PH=7.2] add 10mL thrombocyte or liver homogenate sample, the condition homogenate of 4 ℃ 60 minutes, and the material that will float on the surface is by the filter membrane of 0.2um, the FPLC system of the Pharmacia for tissue (Pharmacia) of soluble part, single passage Q-post wherein contains the EDTA of 1mmol/L and the PMSF solution pre-equilibration of 0.5mmol/L by the buffer solution (PH2.2) of 20mmol/L HEPES, then by the sample of solubility, then in the buffer liquid with 5mL, wash, the PDE isozyme carries out gradient elution with the Nacl (55ml altogether) in identical buffer liquid of 0~500mmol/L different concns, flow velocity 1mL/ minute, pillar is assembled the active part height of PDE, be stored in-80 degrees centigrade standby.
The cyclic nucleotide PDE that isoflavones lignin compound of the present invention and Papaverine are presented in FPLC by isotropic substance (Thomson, the Applemam) experimentation that uses two kinds the activity of PDE5 and PDE4 effect has activity, this mixture reaction (cumulative volume 100ml) comprise post part eluate (10ul-25ul) [
3h]-cGMP/[
3h]-cAMP (0.5umol/L, 2uCi/mL).
Experiment adds radio label substrate or hatching enzyme to put into 30 degrees centigrade of water-baths 20 minutes after starting, sample hose is put to experiment end after 2 minutes in boiling water, the metabolism product is separated with the ion exchange column that metabolism substrate is not processed by AG1-x2 resin and 0.1mol/NaOH wash-out, and determines the catalytic activity of PDE by the radioactivity of flicker pond detection eluate.
Research PDE isozyme restraining effect
In dimethyl sulphoxide liquid, isoflavones lignin compound of the present invention and Papaverine are added in the mixture of temperature bath.The representative reaction that adds of PDE biochemical enzyme starts, whole inhibition experiments are all carried out in the CGMP/CAMP hydrolysis is no more than 15%, and the multiple of enzyme and quantity is with linear growth, the restraining effect research kinetics by isoflavones lignin compound of the present invention, Papaverine to PDE5/PDE4, [
3h]-cGMP/[
3h]-the cAMP concentration of substrate concentrates on 0.3 μ mol/L to 10 μ mol/L, initial percent hydrolysis be at no sample or have under the existence of sample, carry out (10
-10mol/L~10
-4mol/L).
Statistical study
For data, the logarithm of enzymic activity and concentration are made to composes curve, calculate the inhibiting half inhibiting rate of PDE5 s curve as shown in Figure 1, Figure 2.
Test-results:
1. to specific PDE5 activity influence:
Isoflavones lignin compound of the present invention and Papaverine have the restraining effect of dose-dependently to PDE5.
The restraining effect of isoflavones lignin compound of the present invention is 10 at PDE5
-8mol/L (2%)~10
-4mol/L (99.70%).The restraining effect of Papaverine is 10 at PDE5
-8mol/L (4.1%)~10
-4mol/L (96.40%).
Isoflavones lignin compound of the present invention and Papaverine suppress the PDE5 activity and have concentration dependent P<0.01, and the two does not have significant difference;
2. to the PDE4 activity influence:
Isoflavones lignin compound of the present invention and Papaverine have the restraining effect of dose-dependently to PDE4.
Isoflavones lignin compound of the present invention is 10 to the restraining effect of PDE4
-8mol/L (1%)~10
-3mol/L (95%), Papaverine is 10 to the restraining effect of PDE4
-8mol/L (4%)~10
-3mol/L (95%).Isoflavones lignin compound of the present invention and Papaverine have concentration dependent (P<0.01) to the PDE restraining effect, and the restraining effect of isoflavones lignin compound of the present invention is significantly higher than Papaverine.
3.PDE5/PDE4 restraining effect:
Their result can be found out from the S curve
Isoflavones lignin compound of the present invention is 0.43 μ mol/L at the half inhibiting rate of PDE5, and Papaverine is 0.68 μ mol/L.To PDE4 half inhibiting rate concentration, be 0.50 μ mol/L and 3.07 μ mol/L, isoflavones lignin compound of the present invention and the Papaverine selection power (PDE4/PDE5 to PDE5, IC5o) be 1.04 times and 4.72 times, isoflavones lignin compound of the present invention and Papaverine are 0.96 times and 0.21 times to the selection power of PDE4, isoflavones lignin compound of the present invention is Papaverine (PDE4/PDE5, IC to the selectivity inhibiting rate of PDE5
50) 0.22 times, isoflavones lignin compound of the present invention is Papaverine (PDE5/PDE4, IC to the selectivity inhibiting rate of PDE4
50) 4.57 times.
This presentation of results:
Isoflavones lignin compound of the present invention all has restraining effect to PDE4, PDE5, and Papaverine is strong than PDE4 to the PDE5 restraining effect.
The present invention has obtained compound of the present invention from camel thorn plants first, found a kind of new approach for obtaining the isoflavones Lignanoids compounds, the natural drug that plant resources is efficient with exploitation for taking full advantage of, difficulty, increase immunity and antibacterial and anti-inflammation functions are erected in the treatment of low toxicity is significant.
The accompanying drawing explanation:
Fig. 1 is that isoflavones xylogen of the present invention is to the inhibiting half inhibiting rate of PDE5 s graphic representation.
Fig. 2 is that isoflavones xylogen of the present invention is to the inhibiting half inhibiting rate of PDE4 s graphic representation.
Embodiment:
Embodiment 1:
Get the dry former Plant Powder of camel thorn and be broken into the particle of 2 centimetres as raw material, ethanol by every 3500ml 65% adds 10kg raw material refluxing extraction 1.5h, cross 180 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 2 times in the ratio of the ethanol of 10kg material quantity/3000ml 65% again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 2h in the chloroform of 2 times of concentrated solution volumes, cross 100 μ m sieves, trapped substance repeats 2 extractions, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 150 μ m granularities by 8 times of silicagel column cylinder volumes, the sherwood oil that is 15% with acetone content-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary phase is silica gel, granularity 48 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 2:
Get the dry former Plant Powder of camel thorn and be broken into the particle of 1 centimetre as raw material, ethanol by every 3000ml 70% adds 10kg raw material refluxing extraction 2h, cross 150 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 3 times in the ratio of the ethanol of 10kg material quantity/3000ml 65% again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 1.5h in the chloroform of 2 times of concentrated solution volumes, cross 74 μ m sieves, trapped substance repeats 2 extractions, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 124 μ m granularities by 10 times of silicagel column cylinder volumes, the sherwood oil that is 30% with acetone content-acetone mixed solution wash-out, collect elutriant, through ODS high performance liquid phase preparative column, stationary-phase particle size degree 25 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 3:
Get the dry former Plant Powder of camel thorn and be broken into the particle of 0.5 centimetre as raw material, ethanol by every 3000ml 65% adds 10kg raw material refluxing extraction 2h, cross 150 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 2 times in the ratio of the ethanol of 10kg material quantity/2500ml 70% again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 1h in the chloroform of 2.5 times of concentrated solution volumes, cross 104 μ m sieves, trapped substance repeats 2 extractions, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 124 μ m granularities by 10 times of silicagel column cylinder volumes, the sherwood oil that is 50% with acetone content-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 15 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 4:
Getting the dry former plant of camel thorn is cut into 1 centimetre and usings interior section as raw material, ethanol by every 3000ml 65% adds 10kg raw material refluxing extraction 1.5h, cross 74 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 3 times in the ratio of 10kg material quantity/2800ml 65% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 1.5h in the chloroform of 2.5 times of concentrated solution volumes, cross 74 μ m sieves, trapped substance repeats 2 extractions by said ratio, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 74 μ m granularities by 12 times of silicagel column cylinder volumes, with pressing 8 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 25%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 23 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 5:
Getting the dry former plant of camel thorn is cut into 0.5 centimetre and usings interior section as raw material, ethanol by every 3200ml 50% adds 10kg raw material refluxing extraction 1.5h, cross 38 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 3 times in the ratio of 10kg material quantity/2800ml 65% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 1.5h in the chloroform of 2.5 times of concentrated solution volumes, cross 38 μ m sieves, trapped substance repeats 1 extraction by said ratio, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 104 μ m granularities by 9 times of silicagel column cylinder volumes, with pressing 12 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 45%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 13 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 6:
Get the dry former Plant Powder of camel thorn and be broken into particle below 0.8 centimetre as raw material, ethanol by every 3400ml 55% adds 10kg raw material refluxing extraction 1.5h, cross 124 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 3 times in the ratio of 10kg material quantity/2500ml 50% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 1h in the chloroform of 2 times of concentrated solution volumes, cross 44 μ m sieves, trapped substance repeats 2 extractions by said ratio, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 74 μ m granularities by 11 times of silicagel column cylinder volumes, with pressing 8 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 30%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 18 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 7:
Get the dry former Plant Powder of camel thorn and be broken into particle below 1.2 centimetres as raw material, ethanol by every 2500ml 50% adds 10kg raw material refluxing extraction 1.5h, cross 150 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 3 times in the ratio of 10kg material quantity/2500ml 50% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 1.5h in the chloroform of 2 times of concentrated solution volumes, cross 61 μ m sieves, trapped substance is by extraction 1h in the chloroform of 1.5 times of above-mentioned concentrated solution volumes and repeat 2 extractions, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 74 μ m granularities by 10 times of silicagel column cylinder volumes, with pressing 10 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 25%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 10 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 8:
Get the dry former Plant Powder of camel thorn and be broken into particle below 1.0 centimetres as raw material, ethanol by every 3300ml 50% adds 10kg raw material refluxing extraction 1.5h, cross 150 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 3 times in the ratio of 10kg material quantity/2500ml 50% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 1.8h in the chloroform of 2.2 times of concentrated solution volumes, cross 62 μ m sieves, trapped substance is by extraction 1h in the chloroform of 1.5 times of above-mentioned concentrated solution volumes and repeat 2 extractions, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 74 μ m granularities by 8 times of silicagel column cylinder volumes, with pressing 10 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 45%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 6.5 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 9:
Get the dry former Plant Powder of camel thorn and be broken into particle below 0.8 centimetre as raw material, ethanol by every 3500ml 50% adds 10kg raw material refluxing extraction 1.5h, cross 150 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 2 times in the ratio of 10kg material quantity/2800ml 50% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 1.5h in the chloroform of 2.8 times of concentrated solution volumes, cross 38 μ m sieves, trapped substance is by extraction 1.5h in the chloroform of 1.5 times of above-mentioned concentrated solution volumes and repeat 1 extraction, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 58 μ m granularities by 12 times of silicagel column cylinder volumes, with pressing 10 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 30%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 2.6 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 10:
Get the dry former Plant Powder of camel thorn and be broken into particle below 0.5 centimetre as raw material, ethanol by every 3000ml 50% adds 10kg raw material refluxing extraction 2h, cross 150 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 1 time in the ratio of 10kg material quantity/3000ml50% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 2h in the chloroform of 2.5 times of concentrated solution volumes, cross 149 μ m sieves, trapped substance is by extraction 2h in the chloroform of 1.5 times of above-mentioned concentrated solution volumes and repeat 1 extraction, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 106 μ m granularities by 10 times of silicagel column cylinder volumes, with pressing 10 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 30%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 58 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 11:
Get the dry former Plant Powder of camel thorn and be broken into particle below 0.6 centimetre as raw material, ethanol by every 3200ml 50% adds 10kg raw material refluxing extraction 1.2h, cross 124 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 2 times in the ratio of 10kg material quantity/2700ml50% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 2h in the chloroform of 2.2 times of concentrated solution volumes, cross 89 μ m sieves, trapped substance is by extraction 2h in the chloroform of 1.5 times of above-mentioned concentrated solution volumes and repeat 1 extraction, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 74 μ m granularities by 10 times of silicagel column cylinder volumes, with pressing 10 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 45%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 74 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.
Embodiment 12:
Get the dry former Plant Powder of camel thorn and be broken into particle below 0.3 centimetre as raw material, ethanol by every 3300ml 50% adds 10kg raw material refluxing extraction 1.5h, cross 124 μ m aperture sieve, get filtered liquid, trapped substance repeats refluxing extraction 2 times in the ratio of 10kg material quantity/2500ml50% ethanol again, merging filtrate, concentrating under reduced pressure dealcoholysis to alcohol concn≤2% obtains concentrated solution, concentrated solution is extracted to 2h in the chloroform of 2.5 times of concentrated solution volumes, cross 44 μ m sieves, trapped substance is by extraction 1.5h in the chloroform of 1.8 times of above-mentioned concentrated solution volumes and repeat 1 extraction, the gained extraction liquid is concentrated into to chloroform concentration≤2%, measure the silica gel column chromatography of chloroform extraction liquid through 74 μ m granularities by 10 times of silicagel column cylinder volumes, with pressing 10 times of amounts of silicagel column cylinder volume, the sherwood oil that acetone content is 40%-acetone mixed solution wash-out, collect elutriant, through anti-phase ODS high performance liquid phase preparative column, stationary-phase particle size degree 2.5 μ m, gained moving phase is sloughed to sherwood oil and acetone obtain isoflavones Lignanoids compounds of the present invention.