CN101648936A - Chemical composition separation method of traditional Chinese medicine smilax composition and clinical application thereof - Google Patents
Chemical composition separation method of traditional Chinese medicine smilax composition and clinical application thereof Download PDFInfo
- Publication number
- CN101648936A CN101648936A CN200910018907A CN200910018907A CN101648936A CN 101648936 A CN101648936 A CN 101648936A CN 200910018907 A CN200910018907 A CN 200910018907A CN 200910018907 A CN200910018907 A CN 200910018907A CN 101648936 A CN101648936 A CN 101648936A
- Authority
- CN
- China
- Prior art keywords
- composition
- smilax
- ethyl acetate
- compound
- rhamnoside
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Saccharide Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a chemical composition separation method of a traditional Chinese medicine smilax composition and a clinical application thereof. The method comprises the preparation of the water extractive extractum of the traditional Chinese medicine smilax composition, the ethyl acetate extraction of the water extractive material of the traditional Chinese medicine smilax composition,and the separation and measurement of the ethyl acetate extract by silicagel column chromatography. By the method, dihydro-kaempferol, dihydro-kaempferol 3-O-alpha-L-rhamnoside, and 3,3,5,5,7-pentahydroxy-flavanone 3-O-alpha-L-rhamnoside can be obtained, wherein the 3,3,5,5,7-pentahydroxy-flavanone 3-O-alpha-L-rhamnoside is separated from the smilax composition for the first time.
Description
Technical field
The invention belongs to the separation method of medicinal plant chemical ingredients, be specifically related to a kind of chemical composition separation method and clinical application thereof of Chinese medicine smilax composition.
Background technology
Smilax composition mainly adds appropriate amount of auxiliary materials by Chinese medicinal materials chinaroot greenbrier and Rhizome of Glabrous Greenbrier through the medicinal extract of water extract-alcohol precipitation system and makes.The clinical treatment psoriatic that is mainly used in; Though clinical application for many years, curative effect is better, and is unclear always to its pharmaceutical cpd.
Chinaroot greenbrier (Smilax china L.) is a Liliaceae smilax chinaroot greenbrier. root stock; (Leaf of Chinaroot Greenbrier also is used as medicine).
The prior art report, Radix Smilacis Chinae contains smilacin (Smilaxin), Qi glucoside (Isoengeletin) in different, Oleanolic Acid (Oleanolic acid), kaempferol (Kaempferide), dihydro kaempferol (Dihydrokaempferide), β-Gu Zaichun (β-Sitosterol), the β-Gu Zaichun glycoside (β-Sitosteroylglucoside), the former sapogenin A of dioscin (Prosapogenin A of dioscin), dioscin (Dioscin), Taurin (Gracillin), methyl protogracillin (Methylprotogracillin), methyl Protodioscin (Methylprotodioscin) etc.
Its nature and flavor are sweet, temperature; Return sufficient faint the moon, shaoyin channel.Its pharmacological action has diuresis, detoxifcation and anti-trypanosome effect.Function cures mainly, expelling wind and removing dampness, detoxify and promote the subsdence of swelling.Root stock: dispelling the wind and dampness pathogens arthrodynia, wound, gastro-enteritis, dysentery, maldigestion, diabetes, chyluria, leukorrhea, cancer; Leaf: carbuncle furuncle furunculosis is controlled in external application, scalds.
Rhizome of Glabrous Greenbrier (RHIZOMA SMILACIS GLABRAE) is the dry rhizome of liliaceous plant smilacis glabra Smilax glabra Roxb..
The prior art report, Rhizome of Glabrous Greenbrier contains smilax saponin class (Smilax saponins), still contains Tigogein (Tigogenin) and tannin etc.Its nature and flavor are sweet, light, and are flat, return liver, stomach warp; Function with cure mainly dehumidifying, detoxifcation, easing joint movement.Be used for damp and hot stranguria with turbid discharge, band down, carbuncle is swollen, the limbs contraction due to the scrofula, mange, syphilis and mercury poisoning, arthralgia and myalgia.
Summary of the invention
Purpose of the present invention is to separate, measure with silica gel column chromatography to ethyl acetate extract by the ethyl acetate extraction of research Chinese medicine smilax composition the water extracted immersing paste; Explore the chemical active ingredient of smilax composition.
The present invention implements by following technical solution:
Study a kind of chemical composition separation method of Chinese medicine smilax composition, it is characterized in that mainly comprising Chinese medicine smilax composition the water extracted immersing paste preparation, the ethyl acetate extraction of smilax composition the water extracted immersing paste is separated, is measured with silica gel column chromatography to ethyl acetate extract:
(1) smilax composition the water extracted immersing paste preparation
Chinese medicinal materials chinaroot greenbrier and Rhizome of Glabrous Greenbrier mix by 1: 1 weight ratio, add to decoct water gaging decoction three times, 3h for the first time, each 2h of second and third time merges decoction liquor, filter, it is 1.16～1.20/60 ℃ that filtrate is concentrated into relative density, and adding ethanol is 55～60% to containing the alcohol amount, leaves standstill, filter, reclaim ethanol, be condensed into relative density and be 1.27～1.30/60 ℃ smilax composition medicinal extract, standby;
(2) ethyl acetate extraction of smilax composition the water extracted immersing paste
The water extracted immersing paste of smilax composition is dispersed in is hybrid state in the water, carry out ethyl acetate extraction, its weight proportion is:
Smilax composition medicinal extract: ethyl acetate=1: 10;
Water-bath refluxing extraction 2 times, each 1h, extracting solution is concentrated into dried, obtains the ethyl acetate extract of the water extracted immersing paste of smilax composition, and is standby;
(3) silica gel column chromatography to the ethyl acetate extract of the water extracted immersing paste of smilax composition separates, measures to divide and make following steps
1. mix sample
Weight ratio: smilax composition medicinal extract ethyl acetate extract weight: anhydrous silica gel=1: 3
Anhydrous silica gel joins in the smilax composition medicinal extract ethyl acetate extract solution of anhydrous alcohol solution, and heating in water bath volatilizes dehydrated alcohol, and smilax composition medicinal extract ethyl acetate extract sample is adsorbed in the silica gel, and is as gross sample, standby;
2. wet method upper prop
The initial flow phase composite is a volume ratio, ethyl acetate: hexanaphthene=1: 3; Gross sample is mixed thoroughly mutually with the initial flow of moistening amount, adds post footpath 100mm, in the glass chromatography column of the high 1000mm of post;
3. gradient elution under the room temperature
Elution speed is set to 250ml/0.5h, normal pressure, wash-out under the room temperature
Bring into use the initial flow phase, collect elutriant, label is deposited respectively;
Every 250ml collects one bottle, and water-bath refluxes and concentrates, and the inspection of some thin layer plate;
After 59 bottles, change moving phase No. 1: consist of volume ratio, ethyl acetate: hexanaphthene=1: 2;
After 133 bottles, change moving phase No. 2: consist of volume ratio, ethyl acetate: hexanaphthene=1: 1;
After 216 bottles, change moving phase No. 3: consist of volume ratio, ethyl acetate: hexanaphthene=2: 1;
After 248 bottles, change moving phase No. 4: consist of volume ratio, No. 3 moving phases: 95% ethanol=10: 1;
After 275 bottles, change moving phase No. 5: consist of volume ratio, No. 3 moving phases: 95% ethanol=5: 1;
4. crystallization
Detect by thin-layer chromatography, the essentially identical composition of composition is merged, wherein have 221～No. 229 and 230～No. 238 41～No. 68;
After leaving standstill, 41～No. 46 elutriant is separated out light yellow crystal, and suction filtration takes out solid matter, the called after Compound I;
After leaving standstill, 221～No. 229 elutriant is separated out the white powder material, and suction filtration takes out solid matter, called after Compound I I;
After leaving standstill, separate out the white powder material 230～No. 238, suction filtration takes out solid matter, the called after compound III;
5. compound identification
Adopt physicochemical constant, ultraviolet, infrared, hydrogen nuclear magnetic resonance spectrum and nuclear magnetic resonance of carbon spectrum are measured respectively separating obtained compound, determine structure, title;
6. the above-mentioned information of synthesization compound, this separation method obtains dihydrokaempferol, Dihydrokaempferol 3-O-alpha-L-rhamnoside, 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside.Wherein 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside separates from chinaroot greenbrier first and obtains.
A kind of application of Chinese medicine smilax composition is characterized in that the application in preparation treatment psoriatic pharmaceutical preparation, comprises granule, tablet, the application in capsule or the pill.
Advantage of the present invention is:
It is simple and easy that the ethyl acetate extraction of smilax composition the water extracted immersing paste and silica gel column chromatography to ethyl acetate extract separate, measure, and reproducibility is good.
Description of drawings
Fig. 1 is; The infared spectrum of compound Dihydrokaempferol;
Fig. 2 is; The uv-spectrogram of compound Dihydrokaempferol;
Fig. 3 is the compound Dihydrokaempferol
1The H-NMR spectrogram;
Fig. 4 is the compound Dihydrokaempferol
1H-NMR spectrogram (the local amplification);
Fig. 5 is the compound Dihydrokaempferol
1H-NMR spectrogram (the local amplification);
Fig. 6 is the compound Dihydrokaempferol
13The C-NMR spectrogram;
Fig. 7 is the compound Dihydrokaempferol
13C-NMR spectrogram (the local amplification);
Fig. 8 is the compound Dihydrokaempferol
13C-NMR spectrogram (the local amplification);
Fig. 9 is the infared spectrum of compound Dihydrokaempferol 3-O-alpha-L-rhamnoside;
Figure 10 is the uv-spectrogram of compound Dihydrokaempferol 3-O-alpha-L-rhamnoside;
Figure 11 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram;
Figure 12 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
1H-NMR spectrogram (the local amplification);
Figure 13 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
1H-NMR spectrogram (the local amplification);
Figure 14 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
1H-NMR spectrogram (the local amplification);
Figure 15 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
13The C-NMR spectrogram;
Figure 16 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
13C-NMR spectrogram (the local amplification);
Figure 17 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
13C-NMR spectrogram (the local amplification);
Figure 18 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
13C-NMR spectrogram (the local amplification);
Figure 19 is a compound 3,3,5,5, the infared spectrum of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside;
Figure 20 is a compound 3,3,5,5, the uv-spectrogram of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside;
Figure 21 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram;
Figure 22 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
1H-NMR spectrogram (the local amplification);
Figure 23 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
1H-NMR spectrogram (the local amplification);
Figure 24 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
13The C-NMR spectrogram
Figure 25 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
13C-NMR spectrogram (the local amplification);
Figure 26 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
13C-NMR spectrogram (the local amplification);
Figure 27 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
13C-NMR spectrogram (the local amplification).
Embodiment
Below in conjunction with drawings and Examples, the application is further elaborated:
Fig. 1 is the infared spectrum of compound Dihydrokaempferol; Show infrared absorption peak among the figure.
Fig. 2 is the uv-spectrogram of compound Dihydrokaempferol; Show the uv-absorbing bands of a spectrum among the figure.
Fig. 3 is the compound Dihydrokaempferol
1The H-NMR spectrogram; The collection of illustrative plates that shows 0～12ppm section among the figure.
Fig. 4 is that amplify the part of compound Dihydrokaempferol
1The H-NMR spectrogram; The collection of illustrative plates that shows 4.5～7.5ppm section among the figure..
Fig. 5 is that amplify the part of compound Dihydrokaempferol
1The H-NMR spectrogram; The collection of illustrative plates that shows 9.0～12.5ppm section among the figure.
Fig. 6 is the compound Dihydrokaempferol
13The C-NMR spectrogram; The collection of illustrative plates that shows 10～210ppm section among the figure.
Fig. 7 is that amplify the part of compound Dihydrokaempferol
13The C-NMR spectrogram; The collection of illustrative plates that shows 130～220ppm section among the figure.
Fig. 8 is that amplify the part of compound Dihydrokaempferol
13The C-NMR spectrogram; The collection of illustrative plates that shows 65～135ppm section among the figure..
Fig. 9 is the infared spectrum of compound Dihydrokaempferol 3-O-alpha-L-rhamnoside; Show infrared absorption peak among the figure.
Figure 10 is the uv-spectrogram of compound Dihydrokaempferol 3-O-alpha-L-rhamnoside; Show the uv-absorbing bands of a spectrum among the figure.
Figure 11 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram; The collection of illustrative plates that shows 0～12ppm section among the figure.
Figure 12 is that amplify the part of compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram; The collection of illustrative plates that shows 0～3.7ppm section among the figure..
Figure 13 is that amplify the part of compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram; The collection of illustrative plates that shows 4～7.5ppm section among the figure.
Figure 14 is that amplify the part of compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram; The collection of illustrative plates that shows 9.1～12.2ppm section among the figure.
Figure 15 is a compound Dihydrokaempferol 3-O-alpha-L-rhamnoside
13The C-NMR spectrogram; The collection of illustrative plates that shows 10～210ppm section among the figure.
Figure 16 be compound Dihydrokaempferol 3-O-alpha-L-rhamnoside the part amplify
13The C-NMR spectrogram; The collection of illustrative plates that shows 67～82ppm section among the figure..
Figure 17 be compound Dihydrokaempferol 3-O-alpha-L-rhamnoside the part amplify
13The C-NMR spectrogram; The collection of illustrative plates that shows 92～132ppm section among the figure.
Figure 18 be compound Dihydrokaempferol 3-O-alpha-L-rhamnoside the part amplify
13The C-NMR spectrogram; The collection of illustrative plates that shows 155～195ppm section among the figure.
Figure 19 is a compound 3,3,5,5, the infared spectrum of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside; Show infrared absorption peak among the figure.
Figure 20 is a compound 3,3,5,5, the uv-spectrogram of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside; Show the uv-absorbing bands of a spectrum among the figure.
Figure 21 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram; The collection of illustrative plates that shows 0～12.5ppm section among the figure.
Figure 22 is a compound 3,3,5,5, and amplify the part of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram; The collection of illustrative plates that shows 4.1～7.0ppm section among the figure.
Figure 23 is a compound 3,3,5,5, and amplify the part of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
1The H-NMR spectrogram; The collection of illustrative plates that shows 0.6～3.6ppm section among the figure.
Figure 24 is a compound 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
13The C-NMR spectrogram; The collection of illustrative plates that shows 10～210ppm section among the figure..
Figure 25 is a compound 3,3,5,5, and amplify the part of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
13The C-NMR spectrogram; The collection of illustrative plates that shows 140～198ppm section among the figure.
Figure 26 is a compound 3,3,5,5, and amplify the part of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
13The C-NMR spectrogram; The collection of illustrative plates that shows 77～120ppm section among the figure.
Figure 27 is a compound 3,3,5,5, and amplify the part of 7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
13The C-NMR spectrogram; The collection of illustrative plates that shows 65～76ppm section among the figure.
1 reagent and instrument
Reagent name manufacturer or specifications and models
Methyl alcohol (analytical pure) Tianjin Kermel Chemical Reagent Co., Ltd.
Chloroform Laiyang City economic and technological development zone precise treatment factory
Ethyl acetate Yantai three and chemical reagent company limited
Hexanaphthene Tianjin good fortune chemical reagent in morning factory
Dehydrated alcohol Tianjin good fortune chemical reagent in morning factory
Sherwood oil Tianjin good fortune chemical reagent in morning factory
Glacial acetic acid Chemical Reagent Co., Ltd., Sinopharm Group
Methyl alcohol (chromatographically pure)
Instrument title manufacturer or specifications and models
The VARIAN type prepares liquid phase U.S. Varian Inc.
AVANCE-400 type nuclear magnetic resonance analyser Brooker,Switzerland company
ZF-I type ultraviolet lamp Shanghai Gu Cun electric light instrument plant
UV-2550 type ultraviolet spectrophotometer; Day island proper Tianjin company
X-4 type micro melting point apparatus Beijing Tyke Instr Ltd.
DK-98-1 type electric heating constant temperature Shui bath Tianjin Tai Site Instr Ltd.
R-1002N type Rotary Evaporators Zhengzhou Greatwall Scientific Industrial ﹠ Trading Co., Ltd.
The circulation ability of swimming is used vacuum pump Zhengzhou Greatwall Scientific Industrial ﹠ Trading Co., Ltd. more
PE FT1730 infrared chromatograph Perkin-Elmer company
Silica gel column chromatography Yantai chemical industry institute, the 200-300 order; The 100-200 order
Silica gel thin-layer chromatography Yantai Chemical Industry Research Inst.;
Gel filtration chromatography Pharmacia company
Chemical plant, the Changjiang river, polymeric amide Zhengjiang City
Electronic balance AR2130 U.S. Ao Haosi company
2. the selection of developping agent:
Select CHCl according to document and experience
3: EtOAc: MeOH: H
2O=15: 40: 15: 10.
Four kinds of solvents are mixed according to the above ratio, and static 4h takes off layer.
3. smilax composition the water extracted immersing paste preparation
(1) smilax composition the water extracted immersing paste preparation
Chinese medicinal materials chinaroot greenbrier, Rhizome of Glabrous Greenbrier mix by 1: 1 weight ratio, add the convention amount decocting and boil three times, 3h for the first time, each 2h of second and third time merges decoction liquor, filter, it is 1.16～1.20 (60 ℃) that filtrate is concentrated into relative density, and adding ethanol is 55～60% to containing the alcohol amount, leaves standstill, filter, reclaim ethanol, be condensed into relative density and be the medicinal extract of 1.27～1.30 (60 ℃), standby;
4. extract:
Smilax composition medicinal extract is added in the 2000ml round-bottomed flask, add the dosage ethyl acetate,, respectively extracting solution is concentrated into complete drying on Rotary Evaporators, weigh respectively in 100 ℃ of following refluxing extraction twice of water-bath.
A.. mix sample
With anhydrous alcohol solution sample 20g, remain about 1g and make reference substance.
(the sample gross weight: mixed sample in silica gel) 1: 3, and promptly took by weighing 100～200 order anhydrous silica gel 61g, join in the dissolved sample, heating in water bath volatilizes dehydrated alcohol, makes sample powdered, as gross sample with weight ratio.
B. wet method upper prop:
Take by weighing 200～300 order anhydrous silica gel 343g, with ethyl acetate: hexanaphthene=1: 3 is as the initial flow phase.Add gross sample, should slowly add, put one deck absorbent cotton again and protect cylinder.
C. gradient elution:
The about 250ml/30min of elution speed, normal pressure, wash-out under the room temperature (about 20-25 ℃).
Bring into use the initial flow phase, collect elutriant, label is deposited respectively;
Every 250ml collects one bottle, and water-bath refluxes and concentrates, and the inspection of some thin layer plate;
After 59 bottles, change moving phase No. 1: consist of volume ratio, ethyl acetate: hexanaphthene=1: 2;
After 133 bottles, change moving phase No. 2: consist of volume ratio, ethyl acetate: hexanaphthene=1: 1;
After 216 bottles, change moving phase No. 3: consist of volume ratio, ethyl acetate: hexanaphthene=2: 1;
After 248 bottles, change moving phase No. 4: consist of volume ratio, No. 3 moving phases: 95% ethanol=10: 1;
After 275 bottles, change moving phase No. 5: consist of volume ratio, No. 3 moving phases: 95% ethanol=5: 1;
D. crystallization
Detect by thin-layer chromatography, the essentially identical composition of composition is merged, wherein have 221～No. 229 and 230～No. 238 41～No. 68;
After leaving standstill, 41～No. 46 elutriant is separated out light yellow crystal, and suction filtration takes out solid matter, the called after Compound I;
After leaving standstill, 221～No. 229 elutriant is separated out the white powder material, and suction filtration takes out solid matter, called after Compound I I;
After leaving standstill, separate out the white powder material 230～No. 238, suction filtration takes out solid matter, the called after compound III;
E. compound identification
Adopt physicochemical constant, ultraviolet, infrared, hydrogen nuclear magnetic resonance spectrum and nuclear magnetic resonance of carbon spectrum are measured respectively separating obtained compound, determine structure, title;
1.. the structure of Compound I is identified
The crystallization of pale yellow powder shape, is soluble in methyl alcohol, the ethanol equal solvent by mp257-260 ℃.Light green fluorescence is arranged under the UV-light of 365nm wavelength.
UV shows that this compound has two absorption bands, absorption band I maximum absorption wavelength 291.5nm, and absorption band II maximum absorption wavelength 216.5nm can infer tentatively that by the figure spectral shape it is a flavonoid compound.
IR shows that this compound has provided 3432,1635,1517,1450cm
-1Deng absorption peak, can infer hydroxyl in this compound (formation hydrogen bond) thus, carbonyl (conjugation), phenyl ring groups such as (conjugation).
1H-NMR (400MHz, DMSO-d
6) in the collection of illustrative plates, can observe: a hydroxyl proton signal that forms hydrogen bond, δ: 11.91 (1H, s, C
5-OH); Three phenolic hydroxyl group proton signal δ: 10.83 (1H, s, C
7-OH), 9.57 (1H, s, C
4 '-OH), 5.77 (1H, d, C
3-OH); One cover forms A
2B
2The fragrant hydrogen proton signal of coupled system, δ: 7.31 (2H, d, J=8.5Hz, H-2 ' and H-6 '), 6.78 (2H, d, J=8.5Hz, H-3 ' and H-5 '); The fragrant hydrogen proton signal of position between two, δ: 5.91 (1H, d, J=2.04Hz, H-8), 5.85 (1H, d, J=2.04Hz, H-6); In addition can observed hydrogen proton signal: δ: 5.75 (1H, d, J=11.44Hz, H-2), 4.58 (1H, dd, J
1=5.92Hz, J
2=11.44Hz, 3-H).
C-NMR (400MHz DMSO-d6) in the collection of illustrative plates, can observe following carbon signal,
δ：82.85(C-2)，71.43(C-3)，197.86(C-4)，163.28(C-5)，95.99(C-6)，166.76C-7)，94.97(C-8)，162.55(C-9)，100.44(C-1O)，129.42(C-1′)，127.53(C-2′C-6′)，157.71(C-4′)，114.87(C-3′and?C-5′)。
Above spectroscopic data and document
[15]The Dihydrokaempferol data consistent of report is so Compound I I is a Dihydrokaempferol
The above-mentioned figure spectrum information of synthesization compound can judge that this compound is Dihydrokaempferol, spectroscopic data and document
[15]The NMR data consistent of report Dihydrokaempferol.Its structural formula is:
Dihydrokaempferol
2.. the structure of Compound I I is identified
The white powder solid, mp323-325 ℃, be dissolved in methyl alcohol, DMSO is insoluble to acetone, chloroform, ethyl acetate.Light blue fluorescence is arranged under the UV-light of 365nm wavelength.
UV shows that this compound has two absorption bands, absorption band I maximum absorption wavelength 295.5nm, and absorption band II maximum absorption wavelength 217.5nm can infer tentatively that by the figure spectral shape it is a flavonoid compound.
IR shows that this compound has provided 3426,1625,1520,1465cm
-1Deng absorption peak, can infer hydroxyl in this compound (formation hydrogen bond) thus, carbonyl (conjugation), phenyl ring groups such as (conjugation).
1H-NMR (400MHz, DMSO-d
6) in the collection of illustrative plates, can observe: a hydroxyl proton signal that forms hydrogen bond, δ: 11.77 (1H, s, C
5-OH); Two phenolic hydroxyl group proton signal δ: 10.97 (1H, s, C
7-OH), 9.45 (1H, s, C
4 '-OH); One cover forms A
2B
2The fragrant hydrogen proton signal of coupled system, δ: 7.31 (2H, d, J=8.5Hz, H-2 ' and H-6 ') 6.77 (2H, d, J=8.5Hz, H-3 ' and H-5 '); The fragrant hydrogen proton signal of position between two, δ: 5.91 (1H, d, J=2.04Hz, H-8), 5.85 (1H, d, J=2.04Hz, H-6); In addition can observed hydrogen proton signal, δ: 5.61 (1H, d, J=11.44Hz, 2-H), 4.80 (1H, d,, J=11.44Hz, 3-H), conform to substantially with the hydrogen spectrum data of Dihydrokaempferol, wherein the resonance signal of 2-H is compared with Dihydrokaempferol, to low field displacement 0.14ppm, explanation becomes pure glycosides at 3, and coupling constant is 11.4Hz, shows that two protons are two upright reverse couplings.
1H-NMR δ 4.59 (1H, br.s, 1 " H), 1.09 (3H, d, J=6.1Hz, 6-H) and
13The methyl signals of C-NMR δ 17.63 (6-C) is the rhamnosyl characteristic signal.Above spectroscopic data and document
[15]The Dihydrokaempferol 3-O-alpha-L-rhamnoside data consistent of report is so Compound I I is Dihydrokaempferol 3-O-alpha-L-rhamnoside, i.e. engelitin.
(400MHz DMSO-d6) in the collection of illustrative plates, can observe following carbon signal at C-NMR, δ: 80.04 (C-2), 71.12 (C-3), 192.86 (C-4), 164.03 (C-5), 96.21 (C-6), 167.08C-7), 95.16 (C-8), 162.58 (C-9), 100.18 (C-1O), (127.80 C-1 '), (125.53 C-2 ' and C-6 '), 157.25 (C-4 '), 114.83 (C-3 ' and C-5 ').
The above-mentioned figure spectrum information of synthesization compound can judge that this compound is dihydrokaempferol 3-O-alpha-L-rhamnoside, spectroscopic data and a document
[15]The NMR data consistent of report Dihydrokaempferol 3-O-alpha-L-rhamnoside.Its structural formula is:
Dihydrokaempferol 3-O-alpha-L-rhamnoside
3.. the structure of compound III is identified
The white powder solid, mp294-298 ℃, be dissolved in methyl alcohol, DMSO is insoluble to acetone, chloroform, ethyl acetate.Light blue fluorescence is arranged under the UV-light of 365nm wavelength.
UV shows that this compound has two absorption bands, absorption band I maximum absorption wavelength 293.5nm, and absorption band II maximum absorption wavelength 217.5nm can infer tentatively that by the figure spectral shape it is a flavonoid compound.
IR shows that this compound has provided 3436,1622,1521,1461cm
-1Deng absorption peak, can infer hydroxyl in this compound (formation hydrogen bond) thus, carbonyl (conjugation), phenyl ring groups such as (conjugation).
1H-NMR (400MHz, DMSO-d
6) in the collection of illustrative plates, can observe: a hydroxyl proton signal that forms hydrogen bond, δ: 11.77 (1H, s, C
5-OH); Four phenolic hydroxyl group proton signal δ: 10.96,9.47,8.92,8.91; One cover forms AB
2The fragrant hydrogen proton signal of coupled system, δ: 6.49 (2H, s, H-2 ' and H-6 '), 6.85 (1H, s, H-4 '); The fragrant hydrogen proton signal of position between two, δ: 5.96 (1H, d, J=2.04, H-8), 5.92 (1H, d, J=2.04, H-6); Can observe the hydrogen proton signal in addition, δ: 5.61 (1H, d, J=11.44Hz, 2-H), 4.80 (1H, d, J=11.44Hz, 3-H).Wherein the resonance signal of 2-H is compared with Dihydrokaempferol, to low field displacement 0.14ppm, illustrate at 3 to become pure glycosides that coupling constant is 11.4Hz, shows that two protons are two upright reverse couplings.
1H-NMR δ 4.59 (1H, br.s, 1 " H), 1.09 (3H, d, J=6.1Hz, 6-H) and
13The methyl signals of C-NMR δ 17.63 (6-C) is the rhamnosyl characteristic signal.
C-NMR (400MHz, DMSO-d6) in the collection of illustrative plates, can observe following carbon signal, δ:
79.92(C-2)，71.20(C-3)，193.06(C-4)，163.94(C-5)，96.15(C-6)，167.01C-7)，95.15(C-8)，162.45(C-9)，100.28(C-1O)，126.34(C-1′)，114.5(C-2′and?C-6′)，117.56(C-4′)，145.55(C-3′and?C-5′)。
The above-mentioned figure spectrum information of synthesization compound can judge that this compound is 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside, its structure be for:
3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside
4. the above-mentioned information of synthesization compound, this separation method obtains Dihydrokaempferol, Dihydrokaempferol 3-O-alpha-L-rhamnoside, 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside.Wherein 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside separates from smilax composition first and obtains.
Application in preparation treatment psoriatic pharmaceutical preparation comprises granule, tablet, the application in capsule or the pill.
Applicating example: make granule
Chinaroot greenbrier and Rhizome of Glabrous Greenbrier two flavors, boiling three times, 3 hours for the first time, second and third time each 2 hours, collecting decoction, filter, filtrate is concentrated into relative density and is about 1.20, and adding ethanol is 55～60% to containing the alcohol amount, leave standstill, filter, reclaim ethanol, be condensed into relative density and be 1.24～126 medicinal extract.Get 0.8 part of medicinal extract, 3 parts of cane sugar powders, 1 part in dextrin, mixing is made particle, drying, promptly.
All the other formulations are pressed the preparation ordinary method and are got final product.
When treatment,, follow the doctor's advice according to the state of an illness.
Claims (2)
1. the chemical composition separation method of a Chinese medicine smilax composition is characterized in that mainly comprising the preparation of Chinese medicine smilax composition the water extracted immersing paste, and the ethyl acetate extraction of smilax composition the water extracted immersing paste is separated, measured with silica gel column chromatography to ethyl acetate extract:
(1) smilax composition the water extracted immersing paste preparation
Chinese medicinal materials chinaroot greenbrier and Rhizome of Glabrous Greenbrier mix by 1: 1 weight ratio, add to decoct water gaging decoction three times, 3h for the first time, each 2h of second and third time merges decoction liquor, filter, it is 1.16～1.20/60 ℃ that filtrate is concentrated into relative density, and adding ethanol is 55～60% to containing the alcohol amount, leaves standstill, filter, reclaim ethanol, be condensed into relative density and be 1.27～1.30/60 ℃ smilax composition medicinal extract, standby;
(2) ethyl acetate extraction of smilax composition the water extracted immersing paste
The water extracted immersing paste of smilax composition is dispersed in is hybrid state in the water, carry out ethyl acetate extraction, its weight proportion is:
Smilax composition medicinal extract: ethyl acetate=1: 10;
Water-bath refluxing extraction 2 times, each 1h, extracting solution is concentrated into dried, obtains the ethyl acetate extract of the water extracted immersing paste of smilax composition, and is standby;
(3) silica gel column chromatography to the ethyl acetate extract of the water extracted immersing paste of smilax composition separates, measures to divide and make following steps
1. mix sample
Weight ratio: smilax composition medicinal extract ethyl acetate extract weight: anhydrous silica gel=1: 3
Anhydrous silica gel joins in the smilax composition medicinal extract ethyl acetate extract solution of anhydrous alcohol solution, and heating in water bath volatilizes dehydrated alcohol, and smilax composition medicinal extract ethyl acetate extract sample is adsorbed in the silica gel, and is as gross sample, standby;
2. wet method upper prop
The initial flow phase composite is a volume ratio, ethyl acetate: hexanaphthene=1: 3;
Gross sample is mixed thoroughly mutually with the initial flow of moistening amount, adds post footpath 100mm, in the chromatography column of the high 1000mm of post;
3. gradient elution under the room temperature
Elution speed is set to 250ml/0.5h, normal pressure,
Bring into use the initial flow phase, collect elutriant, label is deposited respectively;
Every 250ml collects one bottle, and water-bath refluxes and concentrates, and the inspection of some thin layer plate;
After 59 bottles, change moving phase No. 1: consist of volume ratio, ethyl acetate: hexanaphthene=1: 2;
After 133 bottles, change moving phase No. 2: consist of volume ratio, ethyl acetate: hexanaphthene=1: 1;
After 216 bottles, change moving phase No. 3: consist of volume ratio, ethyl acetate: hexanaphthene=2: 1;
After 248 bottles, change moving phase No. 4: consist of volume ratio, No. 3 moving phases: 95% ethanol=10: 1;
After 275 bottles, change moving phase No. 5: consist of volume ratio, No. 3 moving phases: 95% ethanol=5: 1;
4. crystallization
Detect by thin-layer chromatography, the essentially identical composition of composition is merged, wherein have 221～No. 229 and 230～No. 238 41～No. 68;
After leaving standstill, 41～No. 46 elutriant is separated out light yellow crystal, and suction filtration takes out solid matter, the called after Compound I;
After leaving standstill, 221～No. 229 elutriant is separated out the white powder material, and suction filtration takes out solid matter, called after Compound I I;
After leaving standstill, separate out the white powder material 230～No. 238, suction filtration takes out solid matter, the called after compound III;
5. compound identification
Adopt physicochemical constant, ultraviolet, infrared, hydrogen nuclear magnetic resonance spectrum and nuclear magnetic resonance of carbon spectrum are measured respectively separating obtained compound, determine structure, title;
6. the above-mentioned information of synthesization compound, this separation method obtains dihydrokaempferol, Dihydrokaempferol 3-O-alpha-L-rhamnoside, 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside.Wherein 3,3,5,5,7-penta hydroxy group flavanone 3-O-alpha-L-rhamnoside separates from chinaroot greenbrier first and obtains.
2. the application of a Chinese medicine smilax composition is characterized in that the application in preparation treatment psoriatic pharmaceutical preparation, comprises granule, tablet, the application in capsule or the pill.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910018907XA CN101648936B (en) | 2009-09-17 | 2009-09-17 | Chemical composition separation method of traditional Chinese medicine smilax composition and clinical application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910018907XA CN101648936B (en) | 2009-09-17 | 2009-09-17 | Chemical composition separation method of traditional Chinese medicine smilax composition and clinical application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101648936A true CN101648936A (en) | 2010-02-17 |
| CN101648936B CN101648936B (en) | 2012-08-15 |
Family
ID=41671287
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910018907XA Active CN101648936B (en) | 2009-09-17 | 2009-09-17 | Chemical composition separation method of traditional Chinese medicine smilax composition and clinical application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101648936B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101845467A (en) * | 2010-05-10 | 2010-09-29 | 苏州瑞蓝博中药技术开发有限公司 | Method for extracting kaempferol |
| CN102659853A (en) * | 2012-05-08 | 2012-09-12 | 济南圣泉唐和唐生物科技有限公司 | Method for purifying substances by utilizing chromatographic column |
| CN103239653A (en) * | 2012-02-13 | 2013-08-14 | 陕西兴邦药业有限公司 | Traditional Chinese medicine composition for psoriasis and preparation method thereof |
| CN106492069A (en) * | 2016-12-09 | 2017-03-15 | 南方医科大学南方医院 | Prevent and treat effective ingredient in Chinese of chronic pelvic inflammatory disease and its preparation method and application |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1686521A (en) * | 2005-03-28 | 2005-10-26 | 北京正大绿洲医药科技有限公司 | Psoriasis drip pill and its preparation method |
-
2009
- 2009-09-17 CN CN200910018907XA patent/CN101648936B/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101845467A (en) * | 2010-05-10 | 2010-09-29 | 苏州瑞蓝博中药技术开发有限公司 | Method for extracting kaempferol |
| CN103239653A (en) * | 2012-02-13 | 2013-08-14 | 陕西兴邦药业有限公司 | Traditional Chinese medicine composition for psoriasis and preparation method thereof |
| CN103239653B (en) * | 2012-02-13 | 2014-11-12 | 陕西兴邦药业有限公司 | Traditional Chinese medicine composition for psoriasis and preparation method thereof |
| CN102659853A (en) * | 2012-05-08 | 2012-09-12 | 济南圣泉唐和唐生物科技有限公司 | Method for purifying substances by utilizing chromatographic column |
| CN102659853B (en) * | 2012-05-08 | 2014-10-08 | 济南圣泉唐和唐生物科技有限公司 | Method for purifying substances by utilizing chromatographic column |
| CN106492069A (en) * | 2016-12-09 | 2017-03-15 | 南方医科大学南方医院 | Prevent and treat effective ingredient in Chinese of chronic pelvic inflammatory disease and its preparation method and application |
| CN106492069B (en) * | 2016-12-09 | 2019-10-11 | 南方医科大学南方医院 | Chinese medicine composition and the application for preventing and treating chronic pelvic inflammatory disease |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101648936B (en) | 2012-08-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110455965A (en) | The preparation method and its HPLC fingerprint of pharmaceutical composition | |
| CN101851261B (en) | Polygonum perfoliatum medicinal material, method for preparing reference substance of active constituents in preparation thereof as well as content determination method | |
| Preethi et al. | Screening of natural polysaccharides extracted from the fruits of Pithecellobium dulce as a pharmaceutical adjuvant | |
| CN101648936B (en) | Chemical composition separation method of traditional Chinese medicine smilax composition and clinical application thereof | |
| CN103071006A (en) | Preparation method and quality inspection method of traditional Chinese medicine for treating kidney failure | |
| CN102100818A (en) | Quality control method for lophanthus antifebrile tablets | |
| CN105348192A (en) | Antiviral-activity isoquinoline alkaloid compound in Cassia alata L. and preparation method of antiviral-activity isoquinoline alkaloid compound | |
| Li et al. | Characterization of chemical constituents in Shuanghuanglian oral dosage forms by ultra‐high performance liquid chromatography coupled with time‐of‐flight mass spectrometry | |
| CN108186474A (en) | Loranthus plant Loranthus tanakae extractive of general flavone, preparation method and application | |
| CN104610219B (en) | A kind of xanthones compound with oxidation isopentene group and its preparation method and application | |
| CN105106252A (en) | Method for extracting and separating flavonoid compounds from hypericum japonicum | |
| CN104155371A (en) | HPLC (High Performance Liquid Chromatography) fingerprint spectrum of Chinese herbal medicine compound preparation, namely fluid promoting pills, for treating diabetes and establishing method thereof | |
| CN100388940C (en) | Quality control method of Chinese medicinal preparation for treating child hyperpyrexia | |
| CN101912594A (en) | Traditional Chinese medicine preparation for treating gastrointestinal diseases | |
| CN100542523C (en) | Capsule with hairy vein agrimony and arenobufagin and preparation method thereof, method of quality control | |
| CN102161649A (en) | Polyphenol type derivatives extracted from mulberry leaves and preparation method and applications thereof | |
| CN104257768A (en) | Application of total anthraquinone with various ingredients in stable and uniform proportion and composition of total anthraquinone to treatment of pancreatitis | |
| Reddy et al. | High pressure liquid chromatographic determination of bergenin and (+)‐afzelechin from different parts of Paashaanbhed (Bergenia ligulata yeo) | |
| CN110272459A (en) | Two kinds of noval chemical compounds and its antioxidant activity position in root of Paeonia sinjiangensis | |
| CN104224951A (en) | Total anthraquinones with stable and uniform proportions of various components and composition of total anthraquinones for treating cholecystitis | |
| CN101332216B (en) | Xingsiang Tuerfeng total phenolic acid extract and preparation method thereof | |
| CN112083097B (en) | Thin-layer identification method for simultaneously identifying ferulic acid, calycosin glucoside and hesperidin | |
| CN101647942A (en) | Quality testing method of Yindanxinnaotong soft capsules | |
| CN101284030B (en) | Quality control methods of hairy holly root medicinal materials, extract or hairy holly root preparation | |
| CN1935199A (en) | Quality control method for Chinese medicine compound preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Assignee: Yantai Rongchang Pharmaceutical Co., Ltd. Assignor: RONGCHANG PHARMACEUTICAL (ZIBO) CO., LTD. Contract record no.: 2011370000436 Denomination of invention: Chemical composition separation method of traditional Chinese medicine smilax composition and clinical application thereof License type: Common License Open date: 20100217 Record date: 20110923 |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EC01 | Cancellation of recordation of patent licensing contract |
Assignee: Yantai Rongchang Pharmaceutical Co., Ltd. Assignor: RONGCHANG PHARMACEUTICAL (ZIBO) CO., LTD. Contract record no.: 2011370000436 Date of cancellation: 20120822 |
|
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20100217 Assignee: Yantai Rongchang Pharmacy Co., Ltd. Assignor: RONGCHANG PHARMACEUTICAL (ZIBO) CO., LTD. Contract record no.: 2013370000256 Denomination of invention: Chemical composition separation method of traditional Chinese medicine smilax composition and clinical application thereof Granted publication date: 20120815 License type: Exclusive License Record date: 20131205 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model | ||
| EC01 | Cancellation of recordation of patent licensing contract |
Assignee: Yantai Rongchang Pharmacy Co., Ltd. Assignor: RONGCHANG PHARMACEUTICAL (ZIBO) CO., LTD. Contract record no.: 2013370000256 Date of cancellation: 20141226 |
|
| LICC | Enforcement, change and cancellation of record of contracts on the licence for exploitation of a patent or utility model |