CN106008445A - Flavone and lignin compound and extracting method thereof - Google Patents
Flavone and lignin compound and extracting method thereof Download PDFInfo
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- CN106008445A CN106008445A CN201610317500.7A CN201610317500A CN106008445A CN 106008445 A CN106008445 A CN 106008445A CN 201610317500 A CN201610317500 A CN 201610317500A CN 106008445 A CN106008445 A CN 106008445A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/78—Ring systems having three or more relevant rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/40—Separation, e.g. from natural material; Purification
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Abstract
Elymus nutans is distributed in a large area in China and is one kind of main native herbs in Tibet plateau, and plant resources are abundant. The invention discloses a method for separating 10 flavone and lignin compounds from elymus nutans. The 3 of the 10 compounds are new compounds, and the purity of 8 of the 10 compounds is 95% or above. The compound has multiple physiological effects of liver protection, blood fat reduction, oxidation resistance and the like. In addition, the overground parts of the elymus nutans are utilized, the production cycle is show, less influence is caused on the ecological environment, the high-purity monomeric compound can be quickly prepared, the economic benefits are remarkable, and the method has very important significance on scientific and reasonable development of the elymus nutans resource.
Description
Technical field
The invention belongs to biological pharmacy technical field, be specifically related to a kind of extraction high-purity flavone lignanoids from Elymus nutans
The method of compound.
Background technology
Elymus is a big genus of grass family Tribe Triticeae, has 150 kinds of plants.They all have widely distributed in the whole world, mostly
It is important herbage.China is that this platymiscium is distributed one of relatively broad country.But the chemical constitution study of this platymiscium is the most not
Appear in the newspapers.
Wherein, Elymus nutans has large area to be distributed in China, all has distribution from Northeast plain to Inner Mongolia grassland again to Qinghai-Tibet Platean,
Plant resources enriches.Particularly as one of native ornamental tree that Qinghai-Tibet Platean is main, during long-term evolution, Elymus nutans is
Adapting to Qinghai-Tibet Platean height above sea level, day and night temperature is big, and solar radiation intensity is high, and annual cold and oxygen deficient defines the secondary metabolism of uniqueness
Thing.
According to the literature, flavone Lignanoids compounds have protect the liver, blood fat reducing, antioxidation, prevent diabetes, protection cardiac muscle,
The multiple physiological action such as antiplatelet aggregation and antitumor (progress of Han Baicui, flavone Lignanoids compounds, 2009
Year).This compounds is relatively low at nature relative amount, and this compounds of reported isolated only has 51 at present, distribution
In 20 kinds of plants.
Summary of the invention
This patent provides a kind of 10 flavone method of Lignanoids compounds of separating from Elymus nutans, wherein compound 1,
9 and 10 is noval chemical compound.
The extracting method of this patent, not only enriches the species structure of this compounds, and drapes over one's shoulders alkali for this compounds and vertical fringe
The exploitation of grass provide premise.
The present invention provides the extracting method of flavone lignan compound, comprises the following steps:
Gather Elymus nutans position on the ground, after pulverizing of drying in the shade, with 5-50 times of methanol or ethanol organic solvent extraction, ultrasound wave or
Heating and refluxing extraction 10-90 minute or at ambient temperature merceration, repeat to extract 2-4 time, concentrating under reduced pressure, dry in the shade, obtain the fringe that hangs down
Lyme grass extract；
By extract with after a small amount of aqueous dispersion, being extracted with ethyl acetate, extraction volume ratio is 1:1, extraction times 2-4 time,
Concentrating under reduced pressure, dries in the shade, and obtains ethyl acetate portion；
Ethyl acetate portion is separated by macroporous resin, and with methanol solution gradient elution 3 times, wherein the volume ratio of methanol is successively
For: 30%, 50%, 80%, each 9L；Take 80% methanol fractions concentrating under reduced pressure, dry in the shade and obtain ethyl acetate 80% part；
Ethyl acetate 80% part is separated by MCI resin, with methanol solution gradient elution 7 times, the wherein volume of methanol
Ratio is followed successively by 30%, and 40%, 50%, 60%, 70%, 80%, 90%,；Take 70%-90% part reduced pressure to concentrate, dry in the shade,
To Fr.3；
By the Fr.3 petroleum ether with volume ratio as 2:1, acetone mixed solvent as eluant, use 200-300 mesh analytical pure silica gel,
Carry out silica gel column chromatography；Taking chromatography thing is Fr.3d, and this mixture is the mixture of flavone Lignanoids compounds, concentrating under reduced pressure
Dry in the shade；
Half preparative high-performance liquid chromatographic is utilized to prepare flavone Lignanoids compounds 1-10 from Fr.3d part.
In addition to compound 7 and 8, remaining compound purity is all higher than 95%.Prepare the method detailed in Example of compound 2-8
2-4, the method preparing noval chemical compound 1,9 and 10 is as follows:
Fr.3d 20mL methanol solution is fully vibrated washing, cleaning mixture concentrating under reduced pressure, prepare Fr.3dr, remaining insoluble part
Dry in the shade prepared Fr.3dc.
By Fr.3dr through Sephadex LH-20 column chromatography, with methanol as eluant, remove compound tricin.Remainder
Merge through silica gel column chromatography, after chloroform/methanol=10:1 is eluent, be equipped with by Waters 1525 high performance liquid chromatograph
XBridge C18 chromatographic column (150 × 10mm, 10 μm) (preparation liquid phase below is same instrument), chromatographic solvent is water
And acetonitrile, chromatographic condition is: 25% acetonitrile is isocratic, flow velocity: 2mL/min, detects wavelength: 340nm, prepares new chemical combination
Thing 1 (4mg, retention time: 15.9min).
Taking Fr.3dc and pass through reversed phase chromatography (RP-C18, Mitsubishi Chemical Corporation), eluant is acetone/water (body
Long-pending ratio: 30%, 50%, 70%, 90%), each 2L.Taking 90% part is Fr.3dc3.
Fr.3dc3 part, through Sephadex LH-20 (with chloroform/methanol=1:1 as eluant, 50mL), utilizes thin layer chromatography
Merge similar component, respectively obtain the mixture of compound 9 and 10.The mixture of compound 9 and 10 passes through partly to prepare HPLC,
Chromatographic condition is: 37.5% acetonitrile is isocratic, flow velocity: 2mL/min, detects wavelength: 340nm, prepare noval chemical compound 9 (2mg,
Retention time: 26.7min) and 10 (2mg, retention time: 28.1min).
Elymus nutans is the native ornamental tree being distributed widely in Qinghai-xizang Plateau Region, and plant resources is the abundantest.This patent is utilized
It is the position, ground of this plant, with short production cycle, little to eco-environmental impact, and can quickly prepare high-purity flavone wood fat
Chlorins compound, remarkable in economical benefits, scientific and reasonable exploitation Elymus nutans resource is had very important significance.
This patent prepares gained flavone and Lignanoids compounds technique is simple, and purity is higher, and 3 new chemical combination of isolated first
Thing, enriches the structure type of this compounds, contributes to the deep research and development of this compounds.Additionally, the application of this patent will be
The development and utilization of Qinghai-xizang Plateau Region Elymus nutans provides technical support.
Detailed description of the invention
Below example facilitates a better understanding of the present invention, but does not limit the present invention.Experimental technique in following embodiment, as
Without specified otherwise, it is conventional method.Test material used in following embodiment, if no special instructions, is from routine biochemistry
Reagent Company is commercially available.
Embodiment 1
The flavone of the present invention the preparation method of lignanoids mixture are as follows:
Gather Elymus nutans aerial parts, pulverizing of drying in the shade, obtain fine powder by 100 mesh sieve.Take 3 times of first of 11.8kg fine powder
Alcoholic solution, at room temperature soaks 3 times, is followed successively by 3 days, 3 days, 7 days, and decompression distillation obtains crude extract about 980g.
The water-soluble rear ethyl acetate of crude extract 500mL extracts 3 times successively with volume ratio (1:1), obtains ethyl acetate phase 190g.
Ethyl acetate portion pass through macroporous resin (HP-20, Mitsubishi Chemical Corporation), eluant be methanol-
Water, volume ratio is followed successively by: 30%, and 50%, 80%, methanol, each 9L.Wherein take 80% methanol fractions concentrating under reduced pressure, dry in the shade
To 40g.
Ethyl acetate 80% part passes through MCI resin (CHP 20P, 75～150 μm, Mitsubishi Chemical Corporation)
Separate, with methanol/water (volume ratio is 30%, 40%, 50%, 60%, 70%, 80%, 90%, methanol, each 3L)
Gradient elution.Different component is merged and obtains Fr.1-4 tetra-part, i.e. 30%-40% part be Fr.1,50%-60% be partly Fr.2,
70%-90% part is Fr.3, and it is Fr.4 that methanol rushes post part.Wherein Fr.3 part reduced pressure concentrates, and dries in the shade and obtains 21g.
By Fr.3 part 21g with petroleum ether/acetone (volume ratio as 20:1,10:1,5:1,2:1,1:1, acetone, each 2L) it is
Eluant silicagel column (200-300 mesh analytical pure silica gel) chromatographs.20:1 part be Fr.3a, 10:1 be partly Fr.3b, 5:1 part
It is Fr.3d, 1:1 for Fr.3c, 2:1 part and partial acetone is Fr.3e.Wherein Fr.3d part is flavone Lignanoids compounds
Mixture, concentrating under reduced pressure, dry in the shade and obtain 12g, productivity 0.10%.
Embodiment 2
The preparation method of the compound 1 and 2 of the present invention is as follows:
Take Fr.3d part (12g) 20mL methanol solution fully to vibrate washing, cleaning mixture concentrating under reduced pressure, prepared Fr.3dr (2g),
Remaining part dries in the shade prepared Fr.3dc.
Take Fr.3dr sample segment (0.3g), through Sephadex LH-20 (Sephadex LH-20, Amersham Pharmacia
Biotech) column chromatography, with methanol as eluant (200mL), removes compound tricin (20mg).Remainder merges warp
Silica gel column chromatography, after chloroform/methanol=10:1 is eluent, after utilizing methanol to dissolve, by the efficient liquid phase of Waters 1525
Chromatograph, chromatographic solvent is water and acetonitrile, and chromatographic condition is: 25% acetonitrile is isocratic, flow velocity: 2mL/min, detects wavelength: 340nm,
Accumulation is repeated several times and prepares compound 1 (4mg, retention time: 15.9min) and 2 (2mg, retention time: 17.2min).
Utilize the modern spectral technique such as NMR (Nuclear Magnetic Resonance) spectrum, mass spectrum, infrared and ultraviolet, determine that the chemical formula of compound 1 is
The chemical formula determining compound 2 is
Spectral data is as follows:
Compound 1:(c=0.1, MeOH)；UV(MeOH)λmax272.3,363.9nm；IR(KBr)νmax cm-1:
3421,2924,1653,1567,1513.HRESIMS m/z 501.1152[M+Na]+(calcd for C26H22O9Na,
501.1156)；1H NMR(400MHz,Acetone-d6,δin ppm,J in Hz):δH6.22 (1H, d, J=1.6Hz, H-6),
6.55 (1H, d, J=1.6Hz, H-8), 3.69 (1H, dd, J=10.4,4.4Hz, H-11e), 3.23 (1H, t, J=10.0Hz,
H-11a), 3.55 (1H, dd, J=10.4,4.4Hz, H-12), 4.94 (1H, s, H-13), 6.88 (2H, d, J=8.4Hz, H-15,19),
6.64 (2H, d, J=8.4Hz, H-16,18), 7.51 (1H, s, H-6 '), 3.61 (3H, s, 3 '-OCH3),4.00(3H,s,5′-OCH3),
13.02(1H,s,5-OH)；13C NMR(125MHz,Acetone-d6,δin ppm):δC 159.9(C-2),112.0(C-3),181.5
(C-4),163.2(C-5),99.5(C-6),164.8(C-7),94.7(C-8),158.1(C-9),105.1(C-10),62.6(C-11),42.1
(C-12),36.7(C-13),138.8(C-14),129.2(C-15,19),115.9(C-16,18),156.6(C-17),118.8(C-1′),
128.6(C-2′),147.1(C-3′),148.8(C-4′),144.7(C-5′),103.6(C-6′),60.5(C-3′OMe),56.7(C-5′OMe)。
Compound 2:1H NMR(400MHz,Acetone-d6,δin ppm,J in Hz):δH6.22 (1H, d, J=1.6Hz, H-6), 6.54
(1H, d, J=1.6Hz, H-8), 3.69 (1H, dd, J=10.4,4.4Hz, H-11e), 3.24 (1H, t, J=10.0Hz, H-11a),
3.59 (1H, dd, J=10.4,4.4Hz, H-12), 4.95 (1H, s, H-13), 6.81 (1H, d, J=1.6Hz, H-15), 6.58 (1H, d,
J=1.6Hz, H-18), 6.64 (1H, dd, J=8.0,1.6Hz, H-19), 7.51 (1H, s, H-6 '), 3.14 (3H, s, 16-OCH3),
3.62(3H,s,3′-OCH3),4.00(3H,s,5′-OCH3),13.00(1H,s,5-OH)；13C NMR(125MHz,Acetone-d6,
δin ppm):δC 159.9(C-2),111.0(C-3),181.5(C-4),163.2(C-5),99.5(C-6),164.7(C-7),94.7(C-8),
158.1(C-9),105.1(C-10),62.6(C-11),42.1(C-12),37.1(C-13),136.4(C-14),112.2(C-15),148.2
(C-16),145.9(C-17),115.4(C-18),120.5(C-19),118.9(C-1′),128.4(C-2′),147.1(C-3′),148.8
(C-4′),144.7(C-5′),103.6(C-6′),56.2(C-16OMe),60.5(C-3′OMe),56.8(C-5′OMe)。
Embodiment 3
The preparation method of the compound 3-6 of the present invention is as follows:
Take Fr.3d part (12g) 20mL methanol solution fully to vibrate washing, cleaning mixture concentrating under reduced pressure, prepared Fr.3dr (2g),
Remaining insoluble part is dried in the shade prepared Fr.3dc.
Take Fr.3dc (10g) by anti-phase (RP-C18, Mitsubishi Chemical Corporation), eluant is acetone/water
(volume ratio: 30%, 50%, 70%, 90%), each 2L.30% and 50% part is Fr.3dc1, and 70% part is Fr.3bc2,
90% part is Fr.3dc3, and partial acetone is Fr.3dc4.
Fr.3dc2 part through silica gel column chromatography with chloroform: methanol, volume ratio is followed successively by 20:1,10:1,5:1, each 300mL, ladder
Degree eluting.Taking chloroform: methanol (10:1) part, use 20cm × 20cm preparation silica gel plate, chloroform/methanol=10:1 is eluant,
Obtain two parts, be utilized respectively Sephadex LH-20 column chromatography (with chloroform/methanol=1:1 as eluant, 100mL), purification
Obtain Fr.3dc2-1 and Fr.3dc2-2 two parts.
Taking Fr.3dc2-1 sample segment (50mg), dissolve with methanol, by partly preparing HPLC, chromatographic condition is: 25% acetonitrile
Isocratic, flow velocity: 2mL/min, detects wavelength: 340nm, accumulation is repeated several times and prepares compound 4 (9mg, during reservation
Between: 28.6min) and 3 (9mg, retention time: 30.4min).
Take Fr.3dc2-2 sample segment (60mg), dissolve with methanol, by partly preparing HPLC chromatogram condition be: 25% acetonitrile
Isocratic, flow velocity: 2mL/min, detects wavelength: 340nm, accumulation is repeated several times and prepares compound 5 (10mg, during reservation
Between: 30.4min) and 6 (10mg, retention time: 38.8min).
Utilize the modern spectral technique such as NMR (Nuclear Magnetic Resonance) spectrum, mass spectrum, infrared and ultraviolet, determine compound 3 and the chemistry of compound 4
Formula is:
Utilize the modern spectral technique such as NMR (Nuclear Magnetic Resonance) spectrum, mass spectrum, infrared and ultraviolet, determine compound 5 and the chemistry of compound 6
Formula is:
Spectral data is as follows:
Compound 3:1H NMR(400MHz,DMSO-d6,δin ppm,J in Hz):δH 7.03(1H,s,H-3),6.19(1H,s,H-6),
6.54 (1H, s, H-8), 7.31 (2H, s, H-2 ', 6 '), 7.19 (2H, d, J=8.4Hz, H-2 ", 6 "), 6.68 (2H, d, J=8.4Hz,
H-3 ", 5 "), 4.85 (1H, brs, H-7 "), 4.22 (1H, m, H-8 "), 3.64 (1H, dd, J=11.5,4.4Hz, H-9 " a), 3.23 (1H,
B), dd, J=11.2,4.8Hz, H-9 " 3.86 (6H, s, 3 ', 5 '-OCH3)；13C NMR(100MHz,DMSO-d6,δin ppm):
δC 163.0(C-2),104.9(C-3),181.8(C-4),161.4(C-5),99.2(C-6),164.2(C-7),94.5(C-8),157.5
(C-9),103.7(C-10),125.3(C-1′),104.3(C-2′,6′),153.0(C-3′,5′),140.0(C-4′),132.4(C-1″),127.9
(C-2″,6″),114.4(C-3″,5″),156.3(C-4″),71.6(C-7″),87.1(C-8″),60.4(C-9″),56.4(C-3′,
5′OMe)。
Compound 4:1H NMR(400MHz,DMSO-d6,δin ppm,J in Hz):δH7.02 (1H, s, H-3), 6.19 (1H, d, J=
1.8Hz, H-6), 6.54 (1H, d, J=1.8Hz, H-8), 7.16 (2H, d, J=8.8Hz, H-2 ", 6 "), 6.69 (2H, d, J=8.8
Hz,H-3″,5″),4.77(1H,brs,H-7″),4.31(1H,m,H-8″),3.72(1H,m,H-9″a),3.48(1H,m,H-9″b),
3.86(6H,s,3′,5′-OCH3)；13C NMR(100MHz,DMSO-d6,δin ppm):δC 163.1(C-2),104.9(C-3),
181.9(C-4),161.5(C-5),99.1(C-6),164.3(C-7),94.5(C-8),157.5(C-9),103.9(C-10),125.4(C-1′),
104.3(C-2′,6′),153.1(C-3′,5′),139.4(C-4′),132.6(C-1″),128.0(C-2″,6″),114.5(C-3″,5″),
156.3(C-4″),72.0(C-7″),86.5(C-8″),60.2(C-9″),56.4(C-3′,5′OMe)。
Compound 5:1H NMR(400MHz,Acetone-d6,δin ppm,J in Hz):δH 6.82(1H,s,H-3),6.28(1H,brs,
H-6), 6.58 (1H, brs, H-8), 7.42 (2H, brs, H-2 ', 6 '), 7.07 (1H, d, J=1.4Hz, H-2 "), 6.79 (1H, d, J=
8.1Hz, H-5 "), 6.87 (1H, dd, J=8.1,1.4Hz, H-6 "), 5.05 (1H, d, J=4.4Hz, H-7 "), 4.36 (1H, m,
H-8 "), 3.91 (1H, dd, J=12.0,5.6Hz, H-9 " a), 3.54 (1H, dd, J=12.0,3.2Hz, H-9 " b), 4.01 (6H, s, 3 ',
5′-OCH3),3.84(3H,s,3″-OCH3),12.91(1H,s,5-OH)；13C NMR(100MHz,Acetone-d6,δin ppm):
δC 164.4(C-2),106.0(C-3),183.1(C-4),163.4(C-5),99.9(C-6),165.2(C-7),95.0(C-8),158.9
(C-9),105.4(C-10),127.6(C-1’),105.1(C-2′,6′),154.7(C-3′,5′),140.1(C-4′),133.7(C-1″),111.0
(C-2″),148.0(C-3″),146.6(C-4″),115.2(C-5″),120.2(C-6″),73.6(C-7″),88.2(C-8″),61.2
(C-9″),57.0(C-3′,5′OMe),56.3(C-3″OMe)。
Compound 6:1H NMR(400MHz,Acetone-d6,δin ppm,J in Hz):δH6.82 (1H, s, H-3), 6.28 (1H, d, J=
2.0Hz, H-6), 6.58 (1H, d, J=2.0Hz, H-8), 7.43 (2H, brs, H-2 ', 6 '), 7.07 (1H, d, J=1.6Hz, H-2 "),
6.78 (1H, d, J=8.0Hz, H-5 "), 6.93 (1H, dd, J=8.0,1.6Hz, H-6 "), 5.04 (1H, d, J=7.2Hz, H-7 "),
B), a), 4.18 (1H, m, H-8 "), 3.74 (" 3.39 (1H, dd, J=12.0,3.2Hz, H-9 are " for 1H, dd, J=12.0,3.6Hz, H-9
4.04(6H,s,3′,5′-OCH3),3.82(3H,s,3″-OCH3),12.91(1H,s,5-OH).13C NMR(100MHz,
Acetone-d6,δin ppm):δC 164.6(C-2),106.1(C-3),183.1(C-4),163.4(C-5),99.9(C-6),165.3(C-7),
95.1(C-8),158.9(C-9),105.4(C-10),127.6(C-1’),105.1(C-2′,6′),154.4(C-3′,5′),140.6(C-4′),
133.7(C-1″),111.4(C-2″),148.0(C-3″),146.9(C-4″),115.2(C-5″),120.7(C-6″),73.9(C-7″),89.8
(C-8″),61.8(C-9″),57.0(C-3′,5′OMe),56.2(C-3″OMe)。
Embodiment 4
The preparation method of the compound 7-10 of the present invention is as follows:
Take Fr.3dc3 sample segment (60mg), through Sephadex LH-20 (with chloroform/methanol=1:1 as eluant, 50mL),
Utilize thin layer chromatography to merge similar component, respectively obtain compound 7 and the mixture of 8,9 and 10.
The mixture of compound 7 and 8, dissolves with methanol, and by partly preparing HPLC, chromatographic condition is: 37.5% acetonitrile is isocratic,
Flow velocity: 2mL/min, detects wavelength: 340nm, through being repeated several times that accumulation obtains that ratio is 1:2 and 2:1 7 (retention time:
28.3min) He 8 (retention time: 29.1min) mixture (12mg altogether).
The mixture of compound 9 and 10, dissolves with methanol, and by partly preparing HPLC, chromatographic condition is: 37.5% acetonitrile etc.
Degree, flow velocity: 2mL/min, detects wavelength: 340nm, prepares compound 9 (2mg, reservation through accumulation is repeated several times
Time: 26.7min) and 10 (2mg, retention time: 28.1min).
Utilize the modern spectral technique such as NMR (Nuclear Magnetic Resonance) spectrum, mass spectrum, infrared and ultraviolet, determine compound 7 and the chemistry of compound 8
Formula is:
The chemical formula of compound 9 and compound 10 is:
Spectral data is as follows:
Compound 7:1H NMR(400MHz,Acetone-d6,δin ppm,J in Hz):δH 6.79-6.89(3H,overlapped,H-3,5″,
6 "), 6.28 (1H, br s, H-6), 6.59 (1H, br s, H-8), 7.38 (2H, brs, H-2 ', 6 '), 7.01 (1H, d, J=1.6Hz, H-2 "),
4.61 (1H, d, J=6.8Hz, H-7 "), 4.39 (1H, m, H-8 "), 3.57 (1H, dd, J=12.4,2.8Hz, H-9 " a), 3.28 (1H,
B), dd, J=12.4,4.8Hz, H-9 " 3.99 (6H, s, 3 ', 5 '-OCH3),3.84(3H,s,3″-OCH3),3.18(3H,s,
7″-OCH3),12.92(1H,s,5-OH)；13C NMR(100MHz,Acetone-d6,δin ppm):δC 164.54(C-2),105.8
(C-3),183.0(C-4),163.0(C-5),99.8(C-6),165.2(C-7),95.1(C-8),158.9(C-9),105.4(C-10),126.9
(C-1′),105.1(C-2′,6′),154.5(C-3′,5′),141.7(C-4′),131.0(C-1″),111.8(C-2″),148.2(C-3″),
147.1(C-4″),115.4(C-5″),121.3(C-6″),84.9(C-7″),87.1(C-8″),62.2(C-9″),56.9(C-3′,5′OMe),
56.3(C-3″OMe),57.0(C-7″OMe)。
Compound 8:1H NMR(400MHz,Acetone-d6,δin ppm,J in Hz):δH 6.77-6.88(3H,overlapped,H-3,5″,
6 "), 6.28 (1H, br s, H-6), 6.58 (1H, br s, H-8), 7.34 (2H, brs, H-2 ', 6 '), 7.01 (1H, d, J=1.6Hz, H-2 "),
4.59 (1H, d, J=6.0Hz, H-7 "), 4.34 (1H, m, H-8 "), 3.57 (1H, dd, J=12.8,2.8Hz, H-9 " a), 3.28 (1H,
B), dd, J=12.8,5.2Hz, H-9 " 3.94 (6H, s, 3 ', 5 '-OCH3),3.86(3H,s,3″-OCH3),3.26(3H,s,
7″-OCH3),12.91(1H,s,5-OH)；13C NMR(100MHz,Acetone-d6,δin ppm):δC 164.3(C-2),105.9
(C-3),183.0(C-4),163.0(C-5),99.8(C-6),165.2(C-7),95.1(C-8),158.8(C-9),105.4(C-10),127.4
(C-1′),105.1(C-2′,6′),154.5(C-3′,5′),140.2(C-4′),131.4(C-1″),111.9(C-2″),148.1(C-3″),
147.0(C-4″),115.2(C-5″),121.7(C-6″),83.4(C-7”),86.8(C-8”),60.9(C-9″),56.9(C-3′,5′OMe),
56.3(C-3″OMe),57.2(C-7″OMe)。
Compound 9:UV (MeOH) λmax269.9,340.1nm；HRESIMS m/z 533.1414[M+Na]+(calcd for
C27H26O10Na,533.1418).1H NMR(400MHz,Acetone-d6,δin ppm,J in Hz):δH 6.83(1H,s,overlap,
H-3), 6.28 (1H, br s, H-6), 6.59 (1H, br s, H-8), 7.41 (2H, brs, H-2 ', 6 '), 7.23 (2H, d, J=8.4Hz,
H-2 ", 6 "), 6.84 (2H, d, J=8.4Hz, H-3 ", 5 "), 4.62 (1H, d, J=6.8Hz, H-7 "), 4.38 (1H, m, H-8 "),
B), a), 3.86 (1H, dd, J=11.6,4.0Hz, H-9 " 3.58 (1H, dd, overlapped, H-9 " 4.00 (6H, s, 3 ', 5 '-OCH3),
3.16(3H,s,7″-OCH3),12.92(1H,s,5-OH)；13C NMR(100MHz,Acetone-d6,δin ppm):δC 164.6
(C-2),106.0(C-3),183.1(C-4),163.3(C-5),99.9(C-6),165.2(C-7),96.6(C-8),158.9(C-9),105.4
(C-10),125.4(C-1′),105.1(C-2′,6′),154.5(C-3′,5′),141.7(C-4′),130.4(C-1″),129.8(C-2″,6″),
115.8(C-3″,5″),158.0(C-4″),84.7(C-7″),87.1(C-8″),62.2(C-9″),56.9(C-3′,5′OMe),57.0
(C-7″OMe)。
Compound 10:UV (MeOH) λmax269.9,340.1nm；HRESIMS m/z 511.1596[M+H]+(calcd for
C26H22O9H, 511.1599).1H NMR(400MHz,Acetone-d6,δin ppm,J in Hz):δH 6.77(1H,s,H-3),
6.26 (1H, br s, H-6), 6.55 (1H, br s, H-8), 7.35 (2H, brs, H-2 ', 6 '), 7.24 (2H, d, J=8.4Hz, H-2 ", 6 "),
6.84 (2H, d, J=8.4Hz, H-3 ", 5 "), 4.57 (1H, d, J=6.0Hz, H-7 "), 4.30 (1H, m, H-8 "), 3.86 (1H, dd,
J=12.0,4.0Hz, b), a), H-9 " 3.60 (1H, dd, J=12.0,2.8Hz, H-9 " 3.94 (6H, s, 3 ', 5 '-OCH3),3.24(3H,
s,7″-OCH3),12.90(1H,s,5-OH)；13C NMR(100MHz,DMSO-d6,δin ppm):δC 164.6(C-2),104.7
(C-3),181.6(C-4),163.3(C-5),99.9(C-6),165.2(C-7),96.6(C-8),156.8(C-9),103.2(C-10),125.4
(C-1′),104.1(C-2′,6′),153.0(C-3′,5′),139.0(C-4′),128.8(C-1″),128.6(C-2″,6″),114.7(C-3″,
5″),157.6(C-4″),82.1(C-7″),85.2(C-8″),62.2(C-9″),56.3(C-3′,5′OMe),59.8(C-7″OMe)。
Claims (6)
1. flavone lignan compound, it is characterised in that compound has a following structural formula:
2. flavone the extracting method of lignan compound, it is characterised in that comprise the following steps:
A, collection Elymus nutans position on the ground, after pulverizing of drying in the shade, with 5-50 times of methanol or ethanol organic solvent extraction, ultrasound wave
Or heating and refluxing extraction 10-90 minute or merceration at ambient temperature, repeat to extract 2-4 time, concentrating under reduced pressure, dry in the shade, hung down
Fringe lyme grass extract；
B, by extract with after a small amount of aqueous dispersion, be extracted with ethyl acetate, extraction volume ratio is 1:1, extraction times 2-4
Secondary, concentrating under reduced pressure, dries in the shade, obtains ethyl acetate portion；
C, ethyl acetate portion are separated by macroporous resin, by methanol solution gradient elution 3 times, the wherein volume ratio of methanol
It is followed successively by: 30%, 50%, 80%, takes 80% methanol fractions concentrating under reduced pressure, dry in the shade and obtain ethyl acetate 80% part；
D, ethyl acetate 80% part are separated by MCI resin, with methanol solution gradient elution 7 times, and wherein methanol
Volume ratio is followed successively by 30%, and 40%, 50%, 60%, 70%, 80%, 90%, take 70%-90% part reduced pressure and concentrate, dry in the shade,
Obtain Fr.3；
E, by the Fr.3 petroleum ether with volume ratio as 2:1, acetone mixed solvent as eluant, use 200-300 mesh to analyze pure silicon
Glue, carries out silica gel column chromatography；Taking chromatography thing is Fr.3d, and this mixture is the mixture of flavone Lignanoids compounds, decompression
Concentration is dried in the shade；
F, half preparative high-performance liquid chromatographic is utilized to prepare flavone Lignanoids compounds 1-10 from Fr.3d part.
3. flavone as claimed in claim 2 the extracting method of lignan compound, it is characterised in that described utilization half preparation
High performance liquid chromatography prepares flavone the method for Lignanoids compounds 1 from Fr.3d part, comprises the following steps:
Step one: take Fr.3d part methanol solution and fully vibrate washing, cleaning mixture concentrating under reduced pressure, prepare Fr.3dr；
Step 2: take Fr.3dr sample segment, through Sephadex LH-20 column chromatography, with methanol as eluant, removes chemical combination
Thing tricin；Remainder merges through silica gel column chromatography, after chloroform/methanol=10:1 is eluent, after utilizing methanol to dissolve,
By half preparative high-performance liquid chromatographic instrument, chromatographic solvent is water and acetonitrile, accumulation is repeated several times and prepares compound 1.
4. flavone as claimed in claim 2 the extracting method of lignan compound, it is characterised in that described utilization half system
Standby high performance liquid chromatography prepares flavone Lignanoids compounds 9 and the method for compound 10, including following step from Fr.3d part
Rapid:
Step one: washing that Fr.3d 20mL methanol solution is fully vibrated, cleaning mixture concentrating under reduced pressure, prepare Fr.3dr, remaining
Partly dry in the shade prepared Fr.3dc；
Step 2: taking Fr.3dc and pass through reversed phase chromatography gradient elution 4 times, eluant is volume ratio: 30%, 50%, 70%, 90%
Acetone soln, taking 90% part is Fr.3dc3；
Step 3: Fr.3dc3 part, through Sephadex LH-20 column chromatography, with chloroform/methanol=1:1 as eluant, utilizes thin layer
Chromatography merges similar component, respectively obtains the mixture of compound 9 and 10；The mixture of compound 9 and 10 is by half system
Standby High Performance Liquid Chromatography, chromatographic solvent is water and acetonitrile, prepares compound 9 and 10.
5. flavone as claimed in claim 3 the extracting method of lignan compound, it is characterised in that in described step 2,
High Performance Liquid Chromatography condition is: 25% acetonitrile is isocratic, flow velocity: 2mL/min, detects wavelength: 340nm, noval chemical compound
The retention time of 1 is 15.9min.
6. flavone as claimed in claim 4 the extracting method of lignan compound, it is characterised in that in described step 3,
High Performance Liquid Chromatography condition is: 37.5% acetonitrile is isocratic, flow velocity: 2mL/min, detects wavelength: 340nm, is prepared into
Retention time to noval chemical compound 9: 26.7min, prepares the retention time of noval chemical compound 10: 28.1min.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106436365A (en) * | 2016-10-16 | 2017-02-22 | 饶琼仙 | Production method of gambiered canton gauze discharge print adopting pure natural raw materials |
| CN106436366A (en) * | 2016-10-16 | 2017-02-22 | 饶琼仙 | Production method of discharge prints of gambiered Canton gauze |
| CN108191808A (en) * | 2018-03-27 | 2018-06-22 | 湖南新汇制药股份有限公司 | A kind of compound detached from hedgehog fungus mycelium |
| CN112335792A (en) * | 2019-08-08 | 2021-02-09 | 湖州星座食品有限公司 | Sow feed capable of remarkably increasing survival rate of piglets |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102459217A (en) * | 2009-04-17 | 2012-05-16 | 萨科隆创新有限公司 | Compounds affecting glycemic index |
| CN104447717A (en) * | 2013-09-18 | 2015-03-25 | 北京大学 | Demethylation derivatives of silybi fructus flavonolignan and preparation method and application of demethylation derivatives |
-
2016
- 2016-05-12 CN CN201610317500.7A patent/CN106008445B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102459217A (en) * | 2009-04-17 | 2012-05-16 | 萨科隆创新有限公司 | Compounds affecting glycemic index |
| CN104447717A (en) * | 2013-09-18 | 2015-03-25 | 北京大学 | Demethylation derivatives of silybi fructus flavonolignan and preparation method and application of demethylation derivatives |
Non-Patent Citations (2)
| Title |
|---|
| EVA WENZIG，等: "Flavonolignans from Avena sativa", 《J.NAT.PROD.》 * |
| WU LAN，: "Tricin, a Flavonoid Monomer in Monocot Lignification", 《PLANT PHYSIOLOGY》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106436365A (en) * | 2016-10-16 | 2017-02-22 | 饶琼仙 | Production method of gambiered canton gauze discharge print adopting pure natural raw materials |
| CN106436366A (en) * | 2016-10-16 | 2017-02-22 | 饶琼仙 | Production method of discharge prints of gambiered Canton gauze |
| CN106436366B (en) * | 2016-10-16 | 2019-07-02 | 江苏佳丝缘纺织有限公司 | A kind of Canton gauze gambiered pulls out the production method of print stamp |
| CN108191808A (en) * | 2018-03-27 | 2018-06-22 | 湖南新汇制药股份有限公司 | A kind of compound detached from hedgehog fungus mycelium |
| CN112335792A (en) * | 2019-08-08 | 2021-02-09 | 湖州星座食品有限公司 | Sow feed capable of remarkably increasing survival rate of piglets |
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