CN101380379B - Total flavone in leaves of Murraya paniculata (L.) Jack and preparation method and use thereof - Google Patents
Total flavone in leaves of Murraya paniculata (L.) Jack and preparation method and use thereof Download PDFInfo
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Abstract
The invention provides Murraya paniculata (L.)Jack leaves total flavone, a preparation method and the application thereof. The Murraya paniculata (L.)Jack leaves total flavone is effective part which is extracted and separated from leaves of the Chinese medicine Murraya paniculata (L.)Jack and which takes total flavone as a main component. The total flavone mainly comprises 5,7,3',4'-tetracarboxylic oxygen flavone, 5,7,3',4',5'-pentamethoxyl flavone, 5,6,7,3',4'-pentamethoxyl flavone, 5,6,7,3',4',5'-hexamethoxy flavone and 3-hydroxy-5,7',4'-trimethoxy-flavone. The Murraya paniculata (L.)Jack leaves total flavone has the therapeutic effect on the acute myocardial infarction.
Description
Technical field
The invention belongs to the Natural Medicine Chemistry research field.
Background technology
Murraya paniculata (L.) Jack. [Murraya paniculata (L.) Jack] is Rutaceae Murraya plant, mainly originates from the ground such as Yunnan Province of China, Guizhou, Hunan, Guangdong, Guangxi, Fujian, Taiwan.Being recorded by 2005 editions one one of Pharmacopoeia of People's Republic of China together with another medicinal plants Folium Et Cacumen Murrayae of Murraya paniculata (L.) Jack. and Murraya [Murraya exotica L.], is one of former plant of Folium Et Cacumen Murrayae medical material.
Up to the present having the people to be divided into to get 5 flavone compounds from Murraya paniculata (L.) Jack. [Murraya paniculata (L.) Jack] leaf, is respectively 3,5,6,7,8,3 ', 4 ', 5 '-exoticin (3,5,6,7,8,3 ', 4 ', 5 '-octamethoxyflavone) front-month Fructus Citri tangerinae element (Exotiein); 3,5,7,8,3 ', 4 ', 5 '-Heptamethoxyflavone (3,5,7,8,3 ', 4 ', 5 '-heptamethoxyflavone) be hibiscetin seven methyl ethers (hibiscetin heptamethyl ether); 3,5,6,7,3 ', 4 ', the 5--Heptamethoxyflavone (3,5,6,7,3 ', 4 ', 5 '-heptamethoxyflavone) (the new medical college in Jiangsu, the Chinese medicine voluminous dictionary, the first volume, Shanghai People's publishing house, 1977:44-45); 5,7,8,3 ', 4 ', 5-hexa methoxy flavone (5,7,8,3 ', 4 ', 5 '-hexamethoxyflavone) be Bannamurpanisin (bannamurpanisin) (Botany Gazette 1984,26 (2): 184-186) and 5,7,3 ', 4 ', 5 '-pentamethoxyl flavone (5,7,3 ', 4 ', 5 '-pentamethoxyflavone) (chemical journal 1984,42,1308-1311).
Summary of the invention
The purpose of this invention is to provide a kind of obtain from a thousand li Herba Pelargonii Graveolentis take newfound flavone compound as main component, the mixture with bioactive flavone compound is the Murraya paniculata (L.) Jack. leaf flavonoids, it is a kind of effective ingredient in Chinese.it is characterized in that wherein containing at least 5,7,3 ', 4 '-tetramethoxy flavones, 5,7,3 ', 4 ', 5 '-pentamethoxyl flavone, 5,6,7,3 ', 4 '-pentamethoxyl flavone, 5,6,7,3 ', 4 ', 5 '-hexa methoxy flavone, 7-hydroxyl-5,3 ', 4 '-in trimethoxy flavone two or more, and wherein 5, 7, 3 ', 4 '-tetramethoxy flavones, 5, 7, 3 ', 4 ', 5 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 ', 5 '-the hexa methoxy flavone, 7-hydroxyl-5, 3 ', 4 '-the content sum of trimethoxy flavone is greater than 50% or 5, 7, 3 ', 4 '-tetramethoxy flavones, 5, 7, 3 ', 4 ', 5 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 ', 5 '-the content sum of hexa methoxy flavone is greater than 50% or 5, 7, 3 ', 4 '-tetramethoxy flavones and 5, 7, 3 ', 4 ', 5 '-the content sum of pentamethoxyl flavone is greater than 50% or 5, 7, 3 ', 4 '-content of tetramethoxy flavones is greater than 50%.the present invention also provides the preparation method of Murraya paniculata (L.) Jack. leaf flavonoids, it is characterized in that providing following steps to obtain the Murraya paniculata (L.) Jack. leaf flavonoids: (1) a thousand li Herba Pelargonii Graveolentis decocting boils or the organic solvent reflux, extract, aqueous extract is directly crossed absorption with macroporous adsorbent resin, organic solvent extraction liquid first reclaims organic solvent, rear absorption with macroporous adsorbent resin (2) aqueous alkali excessively or low concentration organic solvent eluting after washing to neutral (3) organic solvent desorbing (4) eluent that is dissolved in water or suspends concentrates, get Murraya paniculata (L.) Jack. leaf extract (5) Murraya paniculata (L.) Jack. leaf extract with water dissolution or cross polyamide column absorption after suspending, the organic solvent eluting, eluent reclaims solvent, obtain Murraya paniculata (L.) Jack. leaf flavonoids (6) Murraya paniculata (L.) Jack. leaf extract or Murraya paniculata (L.) Jack. leaf flavonoids are carried out the higher total flavones of leaf of Folium Et Cacumen Murrayae of column chromatography purification acquisition purity.
Wherein said macroporous adsorbent resin is selected from one or more mixture of AB-8, D4020, D101, D102, D103,860021, HP20; Described column chromatography is selected from one or more of silica gel, aluminium oxide, ODS or polyamide column chromatography; Aqueous slkali is selected from the aqueous solution of alkali metal hydroxide or alkaline earth metal hydroxide.
The present invention also provides the purposes of above-mentioned Murraya paniculata (L.) Jack. leaf flavonoids in preparation treatment myocardial ischemia drug and the pharmaceutical composition of Murraya paniculata (L.) Jack. leaf flavonoids and medically acceptable pharmaceutic adjuvant composition; Its dosage form is tablet, capsule, granule, oral liquid or injection.
The present invention completes by following technical solution.(1) by extraction separation and Structural Identification to the Murraya paniculata (L.) Jack. study on chemical compositions of leaves, found first 5 from a thousand li Herba Pelargonii Graveolentis, 7, 3 ', 4 '-tetramethoxy flavones, 5, 6, 7, 3 ', 4 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 ', 5 '-the hexa methoxy flavone, 7-hydroxyl-5, 3 ', 4 '-trimethoxy flavone and 3 '-hydroxyl-5, 7, 4 '-trimethoxy flavone exist (2) by the research of extracting method, invented with 5, 7, 3 ', 4 '-tetramethoxy flavones, 5, 7, 3 ', 4 ', 5 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 ', 5 '-the hexa methoxy flavone, 3 '-hydroxyl-5, 7, 4 '-trimethoxy flavone is the preparation technology of the Murraya paniculata (L.) Jack. leaf flavonoids of main component.this preparation technology can obtain the Murraya paniculata (L.) Jack. leaf flavonoids (3) of content sum between 50% to 100% of above-mentioned flavone compound and set up 5, 7, 3 ', 4 '-tetramethoxy flavones, 5, 7, 3 ', 4 ', 5 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 ', 5 '-the hexa methoxy flavone, 3 '-hydroxyl-5, 7, 4 '-the HPLC content assaying method (4) of trimethoxy flavone found that by zoopery the medical usage (5) of Murraya paniculata (L.) Jack. leaf flavonoids is studied the pharmaceutical preparation take the Murraya paniculata (L.) Jack. leaf flavonoids as active component, specific as follows.
1, the extraction of Murraya paniculata (L.) Jack. study on chemical compositions of leaves separates
The present invention uses a thousand li Herba Pelargonii Graveolentis available from Yunnan, through being accredited as Rutaceae Murraya plant Murraya paniculata (L.) Jack. [Murraya paniculata (L.) Jack].Decoct with water after a thousand li Herba Pelargonii Graveolentis 2Kg pulverizing with drying and extract 3 times, amount of water is respectively 30 liters, and 24 liters, 20 liters, decocting time is respectively 2 hours, 1.5 hours, 1 hour, merges three times extracting solution.Filtrate is adsorbed by macroporous adsorptive resins, washing, and 95% ethanol desorbing is reclaimed ethanol and is obtained the Murraya paniculata (L.) Jack. leaf extract.With Al on the Murraya paniculata (L.) Jack. leaf extract
2O
3Post is used eluent ethyl acetate, collects eluent, and TLC detects, and merges same composition, obtains A, B, C three parts.A, B, C three parts are carried out silica gel column chromatography repeatedly, obtain flavonoid monomeric compound JA, JB, JC, JD, six monomeric compounds of JE and JF.
the Structural Identification 3 of Murraya paniculata (L.) Jack. study on chemical compositions of leaves ', 4 ', 5 '-pentamethoxyl flavone, 5,6,7,3 ', 4 '-pentamethoxyl flavone, 5,6,7,3 ', 4 ', 5 '-hexa methoxy flavone, 3 '-hydroxyl-5,7,4 '-in trimethoxy flavone two or more, and wherein 5, 7, 3 ', 4 '-tetramethoxy flavones, 5, 7, 3 ', 4 ', 5 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 ', 5 '-the hexa methoxy flavone, 3 '-hydroxyl-5, 7, 4 '-the content sum of trimethoxy flavone is greater than 50% or 5, 7, 3 ', 4 '-tetramethoxy flavones, 5, 7, 3 ', 4 ', 5 '-the pentamethoxyl flavone, 5, 6, 7, 3 ', 4 ', 5 '-the content sum of hexa methoxy flavone is greater than 50% or 5, 7, 3 ', 4 '-tetramethoxy flavones and 5, 7, 3 ', 4 ', 5 '-the content sum of pentamethoxyl flavone is greater than 50% or 5, 7, 3 ', 4 '-content of tetramethoxy flavones is greater than 50%.the present invention also provides the preparation method of Murraya paniculata (L.) Jack. leaf flavonoids, it is characterized in that obtaining as follows the Murraya paniculata (L.) Jack. leaf flavonoids: (1) a thousand li Herba Pelargonii Graveolentis decocting boils or the organic solvent reflux, extract, aqueous extract is directly crossed absorption with macroporous adsorbent resin, organic solvent extraction liquid first reclaims organic solvent, rear absorption with macroporous adsorbent resin (2) aqueous alkali excessively or low concentration organic solvent eluting after washing to neutral (3) organic solvent desorbing (4) eluent that is dissolved in water or suspends concentrates, get Murraya paniculata (L.) Jack. leaf extract (5) Murraya paniculata (L.) Jack. leaf extract with water dissolution or cross polyamide column absorption after suspending, the organic solvent eluting, eluent reclaims solvent, obtain Murraya paniculata (L.) Jack. leaf flavonoids (6) Murraya paniculata (L.) Jack. leaf extract or Murraya paniculata (L.) Jack. leaf flavonoids are carried out the higher Murraya paniculata (L.) Jack. leaf flavonoids of column chromatography purification acquisition purity.
Wherein said macroporous adsorbent resin is selected from one or more mixture of AB-8, D4020, D101, D102, D103,860021, HP20; Described column chromatography is selected from one or more of silica gel, aluminium oxide, ODS or polyamide column chromatography; Aqueous slkali is selected from the aqueous solution of alkali metal hydroxide or alkaline earth metal hydroxide.
The present invention also provides the purposes of above-mentioned Murraya paniculata (L.) Jack. leaf flavonoids in preparation treatment myocardial ischemia drug and the pharmaceutical composition of Murraya paniculata (L.) Jack. leaf flavonoids and medically acceptable pharmaceutic adjuvant composition; Its dosage form is tablet, capsule, granule, oral liquid or injection.
2.1 the Structural Identification of compound JA
JA is white powder, fusing point 195.5-196.5 ℃, show light tone fluorescence under uviol lamp, and be soluble in chloroform, be insoluble in methanol, ethanol, acetone, ethyl acetate, water insoluble.The HCl-Mg reacting positive shows lilac red.FeCl
3Reaction is negative, and prompting is phenolic hydroxy group not, AlCl
3Reacting positive is yellow-green fluorescence under uviol lamp.Above-mentioned response prompting JA may be flavone compound.
13Have 19 signals in CNMR spectrum, composed as can be known by 135 ° and 90 ° of DEPT, wherein δ
C55.70,55.98,56.01 and 56.36 be the methoxyl group carbon signal.
1δ in HNMR
H3.90, the unimodal of 3.94,3.95 places be the methoxyl group proton signal; 7.46 be 1H, dd peak, J=9.0Hz, 1.8Hz, illustrating has adjacent hydrogen on the phenyl ring of parent nucleus, and the position also has a hydrogen between this hydrogen; 6.58 be 1H, s peak, be 3 proton signals; 6.53,6.35 each 1H, be the d peak, J=2.4Hz, illustrate these two hydrogen between being on phenyl ring the position.By the HMBC spectrum as can be known, δ
H(6.35 1H, d, J=2.4Hz) and C
5, C
7, C
8, C
10Relevant, should be attributed to H-6; δ
H(6.53 1H, d, J=2.4Hz) and C
6, C
7, C
9, C
10Relevant, be attributed to H-8; δ
H(6.58 1H, s) and C
2, C
4, C
10, C
1 'Relevant, be attributed to H-3; 6
H(7.26 1H, d, J=1.8Hz) and C
2, C
1 ', C
3 ', C
4 ', C
6 'Relevant, be attributed to H-2 '; δ
H(6.92 1H, d, J=9.0Hz) and C
1 ', C
3 ', C
4 'Relevant, be attributed to H-5 '; δ
H(7.47 1H, dd, J=9.0Hz, 1.8Hz) and C
2, C
2 ', C
4 'Relevant, be attributed to H-6 '; δ
H(3.90 3H, s) and C
7Relevant, be attributed to 7-OCH
3And δ
H3.95 (3H,, s) and C
3 'Relevant, be attributed to 3 '-OCH
3δ
H(3.94 6H, brs) and C
5And C
4 'Relevant, be 4 ' position and 5 absorption signals that methoxyl group is common.It is 343 (M+H) that ES-MS provides molecular ion peak, proves that the JA molecular weight is 342.
According to above analysis, deterministic compound JA is 5,7,3 ', 4 ' ,-tetramethoxy flavones (5,7,3 ', 4 '-tetramethoxyflavone), this compound is got from a thousand li Herba Pelargonii Graveolentis first.
The structural formula of compound JA
Table 1 The
13CNMR and
1HNMR data for compound JA in CDCl
3
[0019]2.2 the Structural Identification of compound JB
JB is white powder, fusing point 199-200 ℃, show light tone fluorescence under uviol lamp, and be soluble in chloroform, be insoluble in methanol, ethanol, ethyl acetate, be insoluble to cold water.The HCl-Mg reacting positive shows lilac red.FeCl
3Reaction negative, prompting may not contain phenolic hydroxyl group.AlCl
3Reacting positive is yellow-green fluorescence under uviol lamp.Above-mentioned response prompting JB may be flavone compound.The Molish reaction negative shows in molecule not sugary.
13Have 17 signals, wherein δ in the CNMR spectrum
C153.43,103.27,56.28 approximately higher 2 times than normal carbon signal, showing has the identical carbon of chemical environment in molecule, namely contain symmetrical structure on the B of parent nucleus ring, and by 135 ° and 90 ° of DEPT as can be known, δ
C153.43 be the quaternary carbon signal, 103.27 is the tertiary carbon signal, 56.28 is the methoxyl group carbon signal, and 60.95,56.36,55.76 is also the methoxyl group signal.
1In HNMR 6
H3.92 (3H), the unimodal methoxyl group proton signal that is located of 3.93 (3H), 3.95 (6H), 3.96 (3H), 7.07 (s, 2H) should be the proton signal on non-adjacent symmetric position on parent nucleus B ring, 6.62 (s, 1H) are 3 proton signals, 6.57 (1H, d, J=2.4Hz), 6.38 (1H, d, J=2.4Hz), be position proton signal between on parent nucleus A ring.Compose as can be known δ by HMBC
H7.07 (2H, s) and C
2, C
1 ', C
3 ', C
4 ', C
6 'Relevant, be attributed to H-2 '; δ
H6.62 (iH, s) and C
2, C
4, C
10, C
1' relevant, be attributed to H-3; δ
H(6.38 d, J=2.4Hz) and C
5, C
7, C
8, C
10Relevant, be attributed to H-6; δ
H(6.57 1H, d, J=2.4Hz) and C
6, C
7, C
9, C
10Relevant, be attributed to H-8; δ
H(3.96 3H, s) and C
5Relevant, be attributed to 5-OCH
3δ
H(3.93 3H, s) and C
7Relevant, be attributed to 7-OCH
3δ
H(3.95 6H, s) and C
3 ', C
5 'Relevant, be attributed to 3 ' and 5 '-OCH
3δ
H(3.92 3H, s) and C
4 'Relevant, be attributed to 4 '-OCH
3It is 373 (M+H) that ES-MS provides molecular ion peak, proves that the JB molecular weight is 372.
According to above analysis, deterministic compound JB is 5,7,3 ', 4 ', 5 '-the pentamethoxyl flavone (5,7,3 ', 4 ', 5 '-pentamethoxyflavone).
The structural formula of compound JB
Table 2 The
13CNMR and
1HNMR data for compound JB in CDCl
3
2.3 the Structural Identification of compound JC
JC is white powder, fusing point 200-201 ℃, show red fluorescence under uviol lamp, and be soluble in chloroform, be insoluble in methanol, ethanol, ethyl acetate, water insoluble.The HCl-Mg reacting positive shows lilac red.FeCl
3Reaction negative, prompting may not contain phenolic hydroxyl group.ALCl
3Reacting positive is yellow-green fluorescence under uviol lamp.Above-mentioned response prompting JC may be flavone compound.The Molish reaction negative shows in molecule not sugary.
13Have 18 signals in the CNMR spectrum, JB compares with compound, and significant change does not occur the chemical displacement value of most of signal, but C
6By δ
C96.13 become 130.52, and at δ
C56.49 how located 1 methoxyl group signal,
1Between having lacked than compound JB in HNMR, digit pair is closed proton signal, δ
H3.97 (3H, s) is the methoxyl group proton signal, this shows that compound JC is the hexa methoxy flavone, and its methoxyl group that has more should be connected on 6 carbon.Compose as can be known δ by HMBC
H6.45 (1H, s) and C
6, C
7, C
9, C
10Relevant, be attributed to H-8; δ
H6.65 (1H, s) and C
2, C
4, C
10, C
1 'Relevant, be attributed to H-3; δ
H(7.19 2H, s) and C
2,, C
1 ', C
3 ', C
4 ', C
6 'Relevant, be attributed to H-2 ' and H-6 '; δ
H(3.92 3H, s) and C
4 'Relevant, be attributed to 4 '-OCH
3δ
H(3.96 6H, s) and C
3 ', C
5 'Relevant, be attributed to 3 ' and 5 '-OCH
3δ
H(3.97 3H, s) and C
5Relevant, be attributed to 5-OCH
3δ
H(4.00 3H, s) and C
6Relevant, be attributed to 6-OCH
3δ
H(4.02 3H, s) and C
7Relevant, be attributed to 7-OCH
3
It is 403 (M+H) that ES-MS provides molecular ion peak, proves that the JC molecular weight is 402.
According to above analysis, deterministic compound JC is 5,6,7,3 ', 4 ', 5 '-the hexa methoxy flavone (5,6,7,3 ', 4 ', 5 '-hexamethoxyflavone).This compound is also for getting from Murraya paniculata (L.) Jack. first.
The structural formula of compound JC
Table 3 The
13CNMR and
1HNMR data for compound JC in CDCl
3
2.4 the Structural Identification of compound JD
JD is white powder, fusing point 207-208 ℃, show light tone fluorescence under uviol lamp, and be soluble in chloroform, be insoluble in methanol, ethanol, ethyl acetate, water insoluble.The HCl-Mg reacting positive shows lilac red.FeCl
3Reacting positive produces blackish green precipitation, and prompting may contain phenolic hydroxyl group.AlCl
3Reacting positive is yellow-green fluorescence under uviol lamp.Above-mentioned response prompting JD may be flavone compound.The Molish reaction negative shows in molecule not sugary.
13Have 18 signals in the CNMR spectrum, composed as can be known by 135 ° of DEPT, wherein δ
C55.87,56.07,56.19 be the methoxyl group carbon signal, other each carbon signal meets the structure of flavone parent nucleus.Because 135 ° of DEPT spectrums show 9 quaternary carbons, and only have three methoxyl groups, illustrating should be with monohydroxy in a certain position.
1δ in HNMR
H3.805, the unimodal of 3.837,3.876 places be the methoxyl group proton signal; 7.46 be 1H, dd peak, J=9.0Hz, 1.8Hz, illustrating has adjacent hydrogen on the phenyl ring of parent nucleus, and the position also has a hydrogen between this hydrogen; 6.67 be 3 proton signals for 1H is unimodal; 6.37,6.57 each 1H, be the d peak, J=2.4Hz, illustrate these two hydrogen between being on phenyl ring the position.By the HMBC spectrum as can be known, δ
H(6.47 1H, d, J=2.4Hz) and C
5, C
7, C
8, C
10Relevant, should be attributed to H-6; δ
H(6.77 1H, d, J=2.4Hz) and C
6, C
7, C
9, C
10Relevant, be attributed to H-8; δ
H(6.58 1H, s) and C
2, C
4, C
10, C
1 'Relevant, be attributed to H-3; δ
H(7.37 1H, d, J=1.8Hz) and C
2, C
3 ', C
4 ', C
6 'Relevant, be attributed to H-2 '; δ
H(7.05 1H, d, J=9.0Hz) and C
1 ', C
3 ', C
4 'Relevant, be attributed to H-5 '; δ
H(7.47 1H, dd, J=9.0Hz, 1.8Hz) and C
2, C
2 ', C
4 'Relevant, be attributed to H-6 '; δ
H3.80 (3H, s) is attributed to 5-OCH
3, δ
H(3.84 3H, s) be attributed to 4 '-OCH
3, δ
H3.87 (3H, s) is attributed to 7-OCH
3It is 329 (M+H) that ES-MS provides molecular ion peak, proves that the JD molecular weight is 328.
According to above analysis, deterministic compound JD is 3 '-hydroxyl-5,7,4 '-trimethoxy flavone. (3 '-hydroxy-5,7,4 '-trimethoxyflavone).This compound is also for getting from Murraya paniculata (L.) Jack. first.
The structural formula of compound JD
Table 4 The
13CNMR and
1HNMR data for compound JD in DMSO
2.5 the structural analysis of compound JE
JE is white powder, fusing point 235-236 ℃, show light tone fluorescence under uviol lamp, and be soluble in chloroform, be insoluble in methanol, ethanol, ethyl acetate, be dissolved in hardly cold water.The HCl-Mg reacting positive shows lilac red.FeCl
3Reacting positive produces blackish green precipitation, and prompting may contain phenolic hydroxyl group.AlCl
3Reacting positive is yellow-green fluorescence under uviol lamp.Above-mentioned response prompting JE may be flavone compound.The Molish reaction negative shows in molecule not sugary.
13Have 18 signals in the CNMR spectrum, composed as can be known by 135 ° of DEPT, wherein δ
C55.99,55.93,55.82 be the methoxyl group carbon signal, other each carbon signal meets the structure of flavone parent nucleus.
1δ in HNMR
H3.786, the unimodal of 3.823,3.855 places be the methoxyl group proton signal; 7.58 be 1H, dd peak, J=9.0Hz, 1.8Hz, illustrating has adjacent hydrogen on the phenyl ring of parent nucleus, and the position also has a hydrogen between this hydrogen; 6.528,1H is unimodal is 3 proton signals; 6.47,6.77 each 1H, be the d peak, J=2.4Hz, illustrate these two hydrogen between being on phenyl ring the position.By the HMBC spectrum as can be known, δ
H(6.37 1H, d, J=2.4Hz) and C
5, C
7, C
8, C
10Relevant, should be attributed to H-6; δ
H(6.57 1H, d, J=2.4Hz) and C
6, C
7, C
9, C
10Relevant, be attributed to H-8; δ
H(6.67 1H, s) and C
2, C
4, C
10, C
1 'Relevant, be attributed to H-3; δ
H(7.47 1H, d, J=1.8Hz) and C
2, C
3 ', C
4 ', C
6 'Relevant, be attributed to H-2 '; δ
H(7.09 1H, d, J=9.0Hz) and C
1 ', C
3 ', C
4 'Relevant, be attributed to H-5 '; δ
H(7.58 1H, dd, J=9.0Hz, 1.8Hz) and C
2, C
2 ', C
4 'Relevant, be attributed to H-6 '; δ
H3.79 (3H, s) is attributed to 5-OCH
3, δ
H3.82 (3H, s) is attributed to 4 ' OCH
3, δ
H(3.86 3H, s) be attributed to 3 '-OCH
3It is 329 (M+H) that ES-MS also provides molecular ion peak, proves that the JE molecular weight is 328.
According to above analysis, deterministic compound JE be 7-hydroxyl-5,3 ', 4 '-trimethoxy flavone.This compound is also for getting from a thousand li Herba Pelargonii Graveolentis first.
The structural formula of compound JE
Table 5 The
13CNMR and
1HNMR data for compound JE in DMSO
*
13The C chemical shift is interchangeable
2.6 the Structural Identification of compound JF
JF is white powder, shows under uviol lamp to be soluble in chloroform by light tone fluorescence, is insoluble in methanol, ethanol, ethyl acetate, is insoluble to cold water.The HCl-Mg reacting positive shows lilac red.FeCl
3Reaction negative, prompting may not contain phenolic hydroxyl group.AlCl
3Reacting positive is yellow-green fluorescence under uviol lamp.Above-mentioned response prompting JB may be flavone compound.The Molish reaction negative shows in molecule not sugary.
13Have 17 signals in CNMR spectrum, by 135 ° and 90 ° of DEPT as can be known, 61.47,56.53,56.25,56.03,55.90 is the methoxyl group signal.
1δ in HNMR
H3.964 (6H), the unimodal methoxyl group proton signal that is located of 3.977 (3H), 3.996 (3H), 4.015 (3H).Compose as can be known 6.444 (s, 1H) and C by HMBC
6, C
7, C
10Relevant, be attributed to 8 proton signals; (6.621 s, 1H) and C
2, C
10, C
2 'Relevant, be attributed to 3 proton signals, 7.424 (1H, d, J=1.8Hz) and C
2, C
3 ', C
4 ', C
6 'Relevant, be attributed to 2 ' position proton signal, 7.597 (1H, dd, J=8.4, J=1.8Hz) and C
2, C
2 ', C
4 'Relevant 6 ' position proton signal, 7.002 (1H, dd, J=8.4) and the C of being attributed to
1 ', C
3 ', C
4 'Relevant, be attributed to 5 ' position proton signal.δ
H(4.015 3H, s) and C
5Relevant, be attributed to 5-OCH
3δ
H(3.996 3H, s) and C
7Relevant, be attributed to 7-OCH
3δ
H(3.964 6H, s) and C
6, C
4 'Relevant, ownership 6 and 4 ' OCH
3δ
H(3.977 3H, s) and C
3 'Relevant, be attributed to 3 '-OCH
3
According to above analysis, deterministic compound JF is 5,6,7,3 ', 4 '-the pentamethoxyl flavone (5,6,7,3 ', 4 '-pentamethoxyflavone).This compound is also for getting from a thousand li Herba Pelargonii Graveolentis first.
The structural formula of compound JF
Table 6 The
13CNMR and
1HNMR data for compound JF in CDCl
3
Compound JA, JB, JC, JD, JE, JF's
1H-NMR,
13C-NMR, DEPT,
1H-
1The spectrograms such as H COSY, HMQC and HMBC are as shown in Fig. 1-37.
3, Murraya paniculata (L.) Jack. leaf flavonoids Study on Preparation
In order to obtain the low total flavones of flavones content high impurity content from a thousand li Herba Pelargonii Graveolentis, the present invention is prepared as follows technique through repeatedly studying finally to have completed.Flavone compound in a thousand li Herba Pelargonii Graveolentis can boil or the method such as organic solvent backflow is extracted by decocting.Because the flavone compound in a thousand li Herba Pelargonii Graveolentis is all low polar compounds, the water boiling and extraction effect is bad, extraction time reaches 7-8 time and could extract fully, therefore preferably adopt organic solvent reflux, extract, method to extract, the reflux, extract, such as concrete available chloroform, ethyl acetate, acetone, methanol, ethanol.Because alcohol toxicity is little, low price, therefore the most handy alcohol reflux.Research finds that therefore the flavone compound in a thousand li Herba Pelargonii Graveolentis can be adopted the absorption with macroporous adsorbent resin method to carry out preliminary purification by absorption with macroporous adsorbent resin, separates the flavones ingredient in extracting solution with impurity such as albumen, polysaccharide, chlorophylls.Aqueous extract can be directly crossed absorption with macroporous adsorbent resin, and organic solvent extraction liquid reclaims solvent thin up to the certain volume, then crosses absorption with macroporous adsorbent resin.Adsorb complete rear low concentration organic solvent or the aqueous slkali eluting first used, purpose is being eluted by the partial impurities of absorption with macroporous adsorbent resin.Wash with water to neutrality after the aqueous alkali eluting is complete, then use the organic solvent desorbing, flavones ingredient is eluted, eluent is concentrated through decompression or normal pressure, gets the Murraya paniculata (L.) Jack. leaf extract.Wherein also contain coumarin and alkaloids composition except flavones ingredient, the content of flavones ingredient can not directly be used for developing new Chinese medicine lower than 50%.Macroporous adsorbent resin described in above-mentioned preparation technology can be selected one or more mixture of AB-8, D4020, D101, D102, D103,860021, HP20 or have the adsorbent resin of other manufacturer's brands of same or similar function; Described column chromatography can be selected one or more of silica gel, aluminium oxide, ODS or polyamide column chromatography.The removal of impurity or eluting can use the mixed solution of the larger organic solvent of methanol, ethanol, acetone, normal propyl alcohol, isopropyl alcohol, n-butyl alcohol isopolarity or they and water with organic solvent, preferably use the aqueous solution of ethanol.When utilizing the aqueous solution eluting removal of impurity of low concentration organic solvent, so-called low concentration is flavone compound is not eluted as the upper limit, generally not higher than 20%.During the removal of impurity of aqueous slkali eluting, operable aqueous slkali has solution or other solution inorganic or organic base of alkali metal hydroxide, alkaline earth metal hydroxide, sodium hydrate aqueous solution preferably, and concentration is advisable with one thousandth.The Murraya paniculata (L.) Jack. leaf extract is crossed polyamide column absorption with water dissolution or after suspending, the organic solvent eluting, eluent reclaims solvent, namely can obtain the Murraya paniculata (L.) Jack. leaf flavonoids, wherein also contain the Coumarins composition except flavones ingredient but the content of flavones ingredient greater than 50%.With said method gained Murraya paniculata (L.) Jack. leaf extract, the rear organic solvent extraction of using that is dissolved in water or suspends, extract concentrating under reduced pressure, drying also can obtain the Murraya paniculata (L.) Jack. leaf flavonoids, the Murraya paniculata (L.) Jack. leaf flavonoids that obtains like this, wherein the content of flavones ingredient increases, and impurity content decreases.The organic solvent that is used for extracting can be chloroform, ethyl acetate, n-butyl alcohol etc. and the not miscible solvent of water, preferably chloroform or ethyl acetate.The Murraya paniculata (L.) Jack. leaf extract that at last said method is obtained or Murraya paniculata (L.) Jack. leaf flavonoids further carry out column chromatography purification and can obtain the higher refining Murraya paniculata (L.) Jack. leaf flavonoids of purity, detect through thin layer chromatography and high performance liquid chromatography, wherein main component is compound JA, JB, JC, JD, JF, their content sum is between 50% to 100%, and can obtain not containing the total flavones of other compositions except flavones ingredient, comprising the Murraya paniculata (L.) Jack. leaf flavonoids that only contains compound JA, JB, JC, JD, JE, JF.Described column chromatography can be selected one or more of silica gel, aluminium oxide, ODS, polyamide column chromatography.
4, the content assaying method of flavone compound research in the Murraya paniculata (L.) Jack. leaf flavonoids
Utilize spectrophotography can measure the total content of flavone compound, but its result does not have the HPLC method accurate, therefore set up the HPLC content assaying method.
(1) instrument, reagent, reference substance
Agilent 1100 Series high performance liquid chromatographs
ZORBAX Extend-C18 4.6 * 250mm ODS, 5 μ m chromatographic columns
The VWD variable wavelength is from external detector
Chromatograph methanol, redistilled water
JA, JB, JC, JD, JF monomer that aforementioned separation is obtained obtain the assay reference substance by recrystallization or ODS purification.By this method chromatographic condition, normalization records purity all over 98%, meets related request.
(2) chromatographic condition
Chromatographic column Zorbax Extend, 4.6 * 250mm, 5 μ m, ODS; Column temperature is 25 ℃; Mobile phase is methanol: water=58: 42; Flow velocity is 1.2mL/min; The detection wavelength is 337nm; Sample size is 20 μ l.
(3) preparation of sample solution
Get dry Folium Et Cacumen Murrayae stem and leaf powder (crossing 60 mesh sieves) 1g, add 78 ℃ of water-bath reflux, extract, of methanol 3 times, add methanol 15ml at every turn, return time is respectively 2h, 1.5h, 1h, merge extractive liquid,, and after solvent evaporated, methanol constant volume is to 50.00ml, filter, get in filtrate 2.0ml to 10ml volumetric flask, standardize solution, and get final product.
(4) assay method
Accurate reference substance liquid and each 20 μ l of need testing solution of inhaling, inject high performance liquid chromatograph respectively, records peak area, utilizes one point external standard method to carry out cubage.
Accompanying drawing 37 compound JF's
1The H-NMR enlarged drawing
The specific embodiment
Get 2 kilograms of a thousand li Herba Pelargonii Graveolentiss, decoct with water and extract 7 times, each amount of water is 24 liters, decocting time is 1 hour, filter, merge extractive liquid, is crossed the AB-8 absorption with macroporous adsorbent resin, the aqueous solution that water washes down 50 liters of one thousandth sodium hydroxide of rear use rinses, wash with water be eluted to thin layer chromatography and can't detect JA with 85% alcoholic solution afterwards to neutrality till.The eluent decompression recycling ethanol gets Murraya paniculata (L.) Jack. leaf extract 153 grams.Detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 23%.Polyamide column absorption will be crossed after this Murraya paniculata (L.) Jack. leaf extract water heating for dissolving, water washes down rear 85% ethanol elution of using, the eluent decompression recycling ethanol, get Murraya paniculata (L.) Jack. leaf flavonoids 74 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is that 51%, TLC shows and wherein also contains the Coumarins composition.Get this Murraya paniculata (L.) Jack. leaf flavonoids and carry out silica gel column chromatography, mixed solution with ethyl acetate and ethanol is that mobile phase is carried out gradient elution, TLC detects, collect the flavone part, reclaim solvent, must make with extra care Murraya paniculata (L.) Jack. leaf flavonoids 35 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 75%, and wherein the content sum of JA, JB, JC is 53%, does not contain other other compositions such as coumarin.
Get 2 kilograms of a thousand li Herba Pelargonii Graveolentiss, 85% alcohol reflux 3 times, amount of alcohol is respectively 10,8,6 times, and return time is 45 minutes, filter, merge extractive liquid, is evaporated to without ethanol flavor, thin up, cross 860021 absorption with macroporous adsorbent resin, till can't detect JA with methanol-eluted fractions to thin layer chromatography after rinsing with 40 liter of 10% ethanol water, eluent reclaim under reduced pressure methanol gets Murraya paniculata (L.) Jack. leaf extract 132 grams.Detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 28%.Polyamide column absorption will be crossed after this Murraya paniculata (L.) Jack. leaf extract water heating for dissolving, water washes down rear 80% ethanol elution of using, the eluent decompression recycling ethanol, get Murraya paniculata (L.) Jack. leaf flavonoids 83 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is that 55%, TLC detects and to show and contain coumarin.Get at last this Murraya paniculata (L.) Jack. leaf flavonoids and carry out alumina column chromatography, mixed solution with ethyl acetate and methanol is that mobile phase is carried out gradient elution, TLC detects, collect the flavone part, reclaim solvent, must make with extra care Murraya paniculata (L.) Jack. leaf flavonoids 38 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 84%, and wherein the content sum of JA, JB is 59%, does not contain the Coumarins composition.
Embodiment 3
Get 2 kilograms of a thousand li Herba Pelargonii Graveolentiss, methanol eddy extracts 3 times, and quantity of methyl alcohol is respectively 10,8,6 times, return time is 1 hour, filter, merge extractive liquid, is evaporated to dried, add the water heating for dissolving, cross the D4020 absorption with macroporous adsorbent resin, water washes down 50 liters of one thousandth potassium hydroxide aqueous solutions of rear use and rinses, wash with water to the neutrality be eluted to thin layer chromatography and can't detect JA with acetone till, eluent reclaim under reduced pressure acetone gets Murraya paniculata (L.) Jack. leaf extract 149 grams.Detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 21%.Polyamide column absorption will be crossed after this Murraya paniculata (L.) Jack. leaf extract water heating for dissolving, water washes down rear 80% methanol-eluted fractions of using, eluent reclaim under reduced pressure methanol, get Murraya paniculata (L.) Jack. leaf flavonoids 89 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is that 58%, TLC detection contains the Coumarins composition.Get at last this Murraya paniculata (L.) Jack. leaf flavonoids and carry out polyamide column chromatography, mixed solution with chloroform and methanol is that mobile phase is carried out gradient elution, TLC detects, collect the flavone part, reclaim solvent, must make with extra care Murraya paniculata (L.) Jack. leaf flavonoids 41 grams, detect through HPLC, wherein the content of JA, JB, JC, JD, JF is 65%, separately contains a small amount of Coumarins composition.
Get 2 kilograms of a thousand li Herba Pelargonii Graveolentiss, acetone reflux, extract, 3 times, amounts of acetone are respectively 10,8,6 times, and return time is 1 hour, filter, merge extractive liquid, is evaporated to driedly, adds the water heating for dissolving, cross the D101 absorption with macroporous adsorbent resin, till after water washes down, 75% isopropyl alcohol was eluted to thin layer chromatography and can't detect JA, eluent reclaim under reduced pressure isopropyl alcohol got Murraya paniculata (L.) Jack. leaf extract 146 grams.Detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 19%.Polyamide column absorption will be crossed after this Murraya paniculata (L.) Jack. leaf extract water heating for dissolving, water washes down rear 80% ethanol elution of using, the eluent decompression recycling ethanol, get Murraya paniculata (L.) Jack. leaf flavonoids 68 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is that 54%, TLC detection contains coumarin.The mixed solution of getting at last these Murraya paniculata (L.) Jack. leaf flavonoids 7 gram second alcohol and waters is that mobile phase is carried out the ODS column chromatography, TLC detects, collect the flavone part, reclaim solvent, must make with extra care Murraya paniculata (L.) Jack. leaf flavonoids 2.9 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 83%, does not contain coumarin.
Get 2 kilograms of a thousand li Herba Pelargonii Graveolentiss, alcohol reflux 3 times, amount of alcohol is respectively 10,8,6 times, return time is 1 hour, filter, merge extractive liquid, is evaporated to dried, add the water heating for dissolving, cross the HP20 absorption with macroporous adsorbent resin, water washes down 50 liters of one thousandth calcium hydroxide aqueous solutions of rear use and rinses, wash with water to the neutrality be eluted to thin layer chromatography and can't detect JA with n-butyl alcohol till, eluent reclaim under reduced pressure n-butyl alcohol gets Murraya paniculata (L.) Jack. leaf extract 172 grams.Detect through HPLC, wherein the content of JA, JB, JC, JD, JF is 18%.To use chloroform extraction after this Murraya paniculata (L.) Jack. leaf extract water heating for dissolving, chloroform layer reclaims solvent, gets Murraya paniculata (L.) Jack. leaf flavonoids 93 grams, detects through HPLC, and wherein the content sum of JA, JB, JC, JD, JF is that 50.5%, TLC detection contains coumarin.Getting at last this Murraya paniculata (L.) Jack. leaf flavonoids is that mobile phase is carried out silica gel column chromatography with the mixed solution of ethyl acetate and ethanol, TLC detects, collect the flavone part, reclaim solvent, must make with extra care Murraya paniculata (L.) Jack. leaf flavonoids 39 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 85%, does not contain coumarin.
Get 2 kilograms of a thousand li Herba Pelargonii Graveolentiss, 85% alcohol reflux 3 times, amount of alcohol is respectively 10,8,6 times, and return time is 45 minutes, filter, merge extractive liquid, is evaporated to without ethanol flavor, thin up, cross 860021 absorption with macroporous adsorbent resin, till can't detect JA with methanol-eluted fractions to thin layer chromatography after rinsing with 60 liter of 10% ethanol water, eluent reclaim under reduced pressure methanol gets Murraya paniculata (L.) Jack. leaf extract 132 grams.Detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 31%.Polyamide column absorption will be crossed after this Murraya paniculata (L.) Jack. leaf extract water heating for dissolving, water washes down rear 80% ethanol elution of using, the eluent decompression recycling ethanol, get Murraya paniculata (L.) Jack. leaf flavonoids 83 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is that 58%, TLC detects and to show and contain coumarin.Get at last this Murraya paniculata (L.) Jack. leaf flavonoids and carry out alumina column chromatography, mixed solution with ethyl acetate and methanol is that mobile phase is carried out gradient elution, TLC detects, collect the flavone part, carrying out purification by silica gel column chromatography after the recovery solvent, must make with extra care Murraya paniculata (L.) Jack. leaf flavonoids 30 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 94%, and wherein the content sum of JA is 51%, does not contain the Coumarins composition.
Embodiment 7
Get 2 kilograms of a thousand li Herba Pelargonii Graveolentiss, decoct with water and extract 7 times, each amount of water is 30 liters, decocting time is 1 hour, filter, merge extractive liquid, is crossed the AB-8 absorption with macroporous adsorbent resin, the aqueous solution that water washes down 70 liters of one thousandth sodium hydroxide of rear use rinses, wash with water be eluted to thin layer chromatography and can't detect JA with 85% alcoholic solution afterwards to neutrality till.The eluent decompression recycling ethanol gets Murraya paniculata (L.) Jack. leaf extract 143 grams.Detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 27%.Polyamide column absorption will be crossed after this Murraya paniculata (L.) Jack. leaf extract water heating for dissolving, water washes down rear 85% ethanol elution of using, the eluent decompression recycling ethanol, get Murraya paniculata (L.) Jack. leaf flavonoids 74 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is that 61%, TLC shows and wherein also contains the Coumarins composition.Get this Murraya paniculata (L.) Jack. leaf flavonoids and carry out silica gel column chromatography, mixed solution with ethyl acetate and ethanol is that mobile phase is carried out gradient elution, and TLC detects, and collects the flavone part, reclaim solvent, must make with extra care Murraya paniculata (L.) Jack. leaf flavonoids 35 grams, with this refining Murraya paniculata (L.) Jack. leaf flavonoids recrystallizing methanol, get Murraya paniculata (L.) Jack. leaf flavonoids 26 grams, detect through HPLC, wherein the content sum of JA, JB, JC, JD, JF is 95%, and wherein the content sum of JA is 53%, does not contain other other compositions such as coumarin.
Get refining Murraya paniculata (L.) Jack. leaf flavonoids 10 grams, add starch appropriate, granulate, incapsulate, make 1000, get Murraya paniculata (L.) Jack. leaf flavonoids capsule.
Embodiment 9
Get Murraya paniculata (L.) Jack. leaf flavonoids 10 grams, add pregelatinized Starch, carboxymethyl starch sodium is appropriate, granulate, tabletting is made 1000, gets Murraya paniculata (L.) Jack. leaf flavonoids sheet.
Embodiment 10
Get refining Murraya paniculata (L.) Jack. leaf flavonoids 10 grams, add 1000ml propylene glycol and 1000ml water for injection, stirring and dissolving is filtered, sterilization, and canned, every 2ml gets Murraya paniculata (L.) Jack. leaf flavonoids injection.
Get Murraya paniculata (L.) Jack. leaf flavonoids 5 grams, add Tween 80 appropriate, add 10 liters of heating for dissolving, high temperature sterilize is distributed into 100 bottles, gets Murraya paniculata (L.) Jack. leaf flavonoids oral liquid.
EXPERIMENTAL EXAMPLE 1
The protective effect research of Murraya paniculata (L.) Jack. leaf flavonoids to expeirmental myocardial ischemia
1 materials and methods
1.1. laboratory animal
The Wistar rat, male and female half and half, body weight 240～260g, the quality certification number are: the moving word 10-5112 of doctor, supplied with by high-new medical faunae research center, Changchun.
1.2. medicine and reagent
1. 2. creatine phosphokinase (CK), lactic acid dehydrogenase (LDH), aspartic acid aminotransferase (AST) reagent are import reagent to Murraya paniculata (L.) Jack. leaf flavonoids (QLXYZHT).
1.3. experimental technique
1. experiment grouping
100 Wistar rats are divided into 5 groups at random, 20 every group, are grouped as follows:
(1) sham operated rats: gavage 0.5% sodium carboxymethyl cellulose 5ml/kg.d * 3d
(2) Infarction Model group: gavage 0.5% sodium carboxymethyl cellulose 5ml/kg.d * 3d
(3) small dose group: gavage Murraya paniculata (L.) Jack. leaf flavonoids 25mg/kg.d * 3d
(4) dosage group in: gavage Murraya paniculata (L.) Jack. leaf flavonoids 50mg/kg.d * 3d
(5) heavy dose of group: gavage Murraya paniculata (L.) Jack. leaf flavonoids 100mg/kg.d * 3d
2. experimental technique and observation index
30min is with Wistar rat etherization after the last administration, facing upward the position is fixed on operating-table, mensuration rat II leads after normal ECG, hair is shaved in the center, do purse string suture with No. 10 line, with scalpel medisection, peel off intercostal muscle with mosquito forceps, 3～4 intercostals are opened breast from the left side, expose heart, find out ramus descendens anterior arteriae coronariae sinistrae between pulmonary conus and left auricle, under left auricle 2～3mm place with No. 0 line ligation immediately it, send heart back to the breast chamber, extrude rapidly gas in the thoracic cavity, tension purse string suture toe-in pricks to close the breast chamber, wholely open the breast time and be no more than 30 seconds, not ligation of sham operated rats and only put surgical thread.After ligation arteria coronaria 24h, with the anesthesia of pentobarbital sodium 30mg/kg abdominal vein, every group of half animal abdominal aortic cannulation got blood, surveys serum CK, AST and LDH with the COBAS-FARA automatic biochemistry analyzer active.Get to cut open after blood and get rat heart, clean hematocele in the chambers of the heart with normal saline, remove atrial tissue and fat, weigh, with 4～5 of myocardium of left ventricle crosscuts, then immerse in the N-BT phosphate buffer, put 37 ℃ of waters bath with thermostatic control, take out after dyeing fully, normal structure dyeing, ischemic tissue does not dye.The cutting-out ischemic myocardium is weighed, with the percentage calculation myocardial infarction area (MIS) of ischemic myocardium and left ventricle weight in wet base.
3. statistical method
Experimental data is processed with SPSS 10.0 statistical softwares, and (x ± s) expression is relatively adopted variance analysis between many groups, relatively adopts the t check between two groups with mean ± standard deviation.
2 results
2.1. the Murraya paniculata (L.) Jack. leaf flavonoids is to rats with acute myocardial infarction MIS, the impact of serum CK, AST and LDH activity
Compare with sham operated rats, Infarction Model group MIS, serum CK, AST and LDH activity all obviously increase (P＜0.05～P＜0.001), show that ami model builds up.Compare with the Infarction Model group, Murraya paniculata (L.) Jack. leaf flavonoids 25,50,100mg/kg group all can obviously be dwindled MIS, reduce serum CK, AST and LDH active (P＜0.05 or P＜0.01), the 100mg/kg group has no significant effect (P＞0.05) to These parameters, sees Table 1.
Tab.1 Effects of QLXYZHT on MIS and serum CK，AST and LDH
activities in acute myocardial infarct rats(x±s，n＝10)
*P＜0.05，**P＜0.01，***P＜0.001 compared with model.
Experimental result shows, the Murraya paniculata (L.) Jack. leaf flavonoids has the effect for the treatment of acute myocardial infarction.
Description of drawings
Accompanying drawing 1 compound JA's
1H-NMR
Accompanying drawing 2 compound JA's
13C-NMR
The DEPT spectrum of accompanying drawing 3 compound JA
Accompanying drawing 4 compound JA's
1H-
1H COSY
The HMQC of accompanying drawing 5 compound JA
The HMBC of accompanying drawing 6 compound JA
Accompanying drawing 7 compound JB's
1H-NMR
Accompanying drawing 8 compound JB's
13C-NMR
The DEPT spectrum of accompanying drawing 9 compound JB
Accompanying drawing 10 compound JB's
1H-
1H COSY
The HMQC of accompanying drawing 11 compound JB
The HMBC of accompanying drawing 12 compound JB
Accompanying drawing 13 compound JC's
1H-NMR
Accompanying drawing 14 compound JC's
13C-NMR
The DEPT spectrum of accompanying drawing 15 compound JC
Accompanying drawing 16 compound JC's
1H-
1H COSY
The HMQC of accompanying drawing 17 compound JC
The HMBC of accompanying drawing 18 compound JC
Accompanying drawing 19 compound JD's
1H-NMR
Accompanying drawing 20 compound JD's
13C-NMR
The DEPT spectrum of accompanying drawing 21 compound JD
Accompanying drawing 22 compound JD's
1H-
1H COSY
The HMQC of accompanying drawing 23 compound JD
The HMBC of accompanying drawing 24 compound JD
Accompanying drawing 25 compound JE's
1H-NMR
Accompanying drawing 26 compound JE's
13C-NMR
The DEPT spectrum of accompanying drawing 27 compound JE
Accompanying drawing 28 compound JE's
1H-
1H COSY
The HMQC of accompanying drawing 29 compound JE
The HMBC of accompanying drawing 30 compound JE
Accompanying drawing 31 compound JF's
1H-NMR
Accompanying drawing 32 compound JF's
13C-NMR
The DEPT spectrum of accompanying drawing 33 compound JF
Accompanying drawing 34 compound JF's
1H-
1H COSY
The HMQC of accompanying drawing 35 compound JF
The HMBC of accompanying drawing 36 compound JF
Claims (12)
1. Murraya paniculata (L.) Jack. leaf flavonoids is characterized in that boiling or organic solvent reflux, extract, a thousand li Herba Pelargonii Graveolentis obtains by decocting, and wherein 5,7,3 ', 4 '-tetramethoxy flavones, 5,7,3 ', 4 ', 5 '-pentamethoxyl flavone, 5,6,7,3 ', 4 '-pentamethoxyl flavone, 5,6,7,3 ', 4 ', 5 '-hexa methoxy flavone, 7-hydroxyl-5,3 ', 4 '-the content sum of trimethoxy flavone is greater than 50%.
2. Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 1, it is characterized in that wherein 5,7,3 ', 4 '-tetramethoxy flavones, 5,7,3 ', 4 ', 5 '-pentamethoxyl flavone, 5,6,7,3 ', 4 ', 5 '-the content sum of hexa methoxy flavone is greater than 50%.
3. Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 1, it is characterized in that wherein 5,7,3 ', 4 '-tetramethoxy flavones and 5,7,3 ', 4 ', 5 '-the content sum of pentamethoxyl flavone is greater than 50%.
4. Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 1, it is characterized in that wherein 5,7,3 ', 4 '-content of tetramethoxy flavones is greater than 50%.
5. Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 1, is characterized in that wherein not containing the Coumarins composition.
6. the preparation method of Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 1, it is characterized in that obtaining as follows corresponding Murraya paniculata (L.) Jack. leaf flavonoids: (1) a thousand li Herba Pelargonii Graveolentis decocting boils or the organic solvent reflux, extract, aqueous extract is directly crossed absorption with macroporous adsorbent resin, organic solvent extraction liquid first reclaims organic solvent, rear absorption with macroporous adsorbent resin (2) aqueous alkali or the low concentration organic solvent eluting crossed is dissolved in water or suspends, be washed to neutrality (3) organic solvent desorbing (4) eluent concentrated, get Murraya paniculata (L.) Jack. leaf extract (5) Murraya paniculata (L.) Jack. leaf extract with water dissolution or cross polyamide column absorption after suspending, the organic solvent eluting, eluent reclaims solvent, obtain Murraya paniculata (L.) Jack. leaf flavonoids (6) Murraya paniculata (L.) Jack. leaf extract or Murraya paniculata (L.) Jack. leaf flavonoids are carried out the higher Murraya paniculata (L.) Jack. leaf flavonoids of column chromatography purification acquisition purity.
7. the preparation method of Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 6 is characterized in that described macroporous adsorbent resin is selected from a kind of of AB-8, D4020, D101, D102, D103,860021, HP20 or uses their mixture.
8. the preparation method of Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 6, is characterized in that described column chromatography is selected from one or more of silica gel, aluminium oxide, ODS or polyamide column chromatography.
9. the preparation method of Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 6, is characterized in that described aqueous slkali is selected from the aqueous solution of alkali metal hydroxide or alkaline earth metal hydroxide.
10. the purposes of Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 1 in preparation prevention and treatment myocardial ischemia drug.
11. the pharmaceutical composition that Murraya paniculata (L.) Jack. leaf flavonoids claimed in claim 1 and medically acceptable pharmaceutic adjuvant form.
12. the described pharmaceutical composition of claim 11 is characterized in that its dosage form is selected from tablet, capsule, granule, oral liquid or injection.
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| CN102669195A (en) * | 2012-06-07 | 2012-09-19 | 海南大学 | Method for extracting total flavonoids from murraya paniculata stems and leaves |
| CN102696699A (en) * | 2012-06-07 | 2012-10-03 | 海南大学 | Murraya paniculata (L.) total flavones soluble powder and preparation method thereof |
| CN102697770A (en) * | 2012-06-13 | 2012-10-03 | 长春瑞德医药科技有限公司 | Murraya paniculata leaf and application of mono compound in preparation of sedative and hypnotic drugs |
| CN103446294B (en) * | 2013-09-18 | 2015-06-24 | 海南华拓天涯制药有限公司 | Murrayae folium ET Cacumen extract preparation method, Murrayae folium ET Cacumen extract obtained thereby and application thereof |
| CN111198235B (en) * | 2020-01-13 | 2021-05-11 | 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 | Method for detecting content of isosinensetin in plasma |
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