CN112194690B - 3 compounds in radix Rubiae and extraction and separation method - Google Patents
3 compounds in radix Rubiae and extraction and separation method Download PDFInfo
- Publication number
- CN112194690B CN112194690B CN202011078958.4A CN202011078958A CN112194690B CN 112194690 B CN112194690 B CN 112194690B CN 202011078958 A CN202011078958 A CN 202011078958A CN 112194690 B CN112194690 B CN 112194690B
- Authority
- CN
- China
- Prior art keywords
- column chromatography
- compound
- compounds
- components
- extraction
- 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.)
- Active
Links
- 150000001875 compounds Chemical class 0.000 title claims abstract description 27
- 238000000605 extraction Methods 0.000 title claims abstract description 13
- 238000000926 separation method Methods 0.000 title claims abstract description 11
- RRHGJUQNOFWUDK-UHFFFAOYSA-N Isoprene Chemical compound CC(=C)C=C RRHGJUQNOFWUDK-UHFFFAOYSA-N 0.000 claims abstract description 4
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 claims abstract description 3
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- 241000123069 Ocyurus chrysurus Species 0.000 claims abstract 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 60
- 238000004440 column chromatography Methods 0.000 claims description 22
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 17
- 238000010898 silica gel chromatography Methods 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- 238000004809 thin layer chromatography Methods 0.000 claims description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 12
- 229940126214 compound 3 Drugs 0.000 claims description 10
- 238000010828 elution Methods 0.000 claims description 10
- UREBWPXBXRYXRJ-UHFFFAOYSA-N ethyl acetate;methanol Chemical compound OC.CCOC(C)=O UREBWPXBXRYXRJ-UHFFFAOYSA-N 0.000 claims description 10
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 claims description 10
- 229940125782 compound 2 Drugs 0.000 claims description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- 229940125904 compound 1 Drugs 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 239000003208 petroleum Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical group O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- 125000001033 ether group Chemical group 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims 1
- 238000013375 chromatographic separation Methods 0.000 claims 1
- 238000010992 reflux Methods 0.000 claims 1
- 239000000741 silica gel Substances 0.000 claims 1
- 229910002027 silica gel Inorganic materials 0.000 claims 1
- 239000000463 material Substances 0.000 abstract description 8
- 238000011160 research Methods 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 6
- 238000001228 spectrum Methods 0.000 abstract description 5
- 238000001819 mass spectrum Methods 0.000 abstract description 3
- 239000000178 monomer Substances 0.000 abstract description 3
- NIFMQAWFYKQXFE-GARIZIQBSA-N OC=1C(C(C=C2C(C3=CC(=CC=C3C(C=12)=O)O)=O)(O[C@]1(O)[C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)O)CO Chemical compound OC=1C(C(C=C2C(C3=CC(=CC=C3C(C=12)=O)O)=O)(O[C@]1(O)[C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)O)CO NIFMQAWFYKQXFE-GARIZIQBSA-N 0.000 abstract description 2
- 230000000857 drug effect Effects 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000003908 quality control method Methods 0.000 abstract description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 45
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 12
- 238000005160 1H NMR spectroscopy Methods 0.000 description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 6
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 5
- 241001149655 Rubia tinctorum Species 0.000 description 5
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 4
- 238000012306 spectroscopic technique Methods 0.000 description 4
- 241001103643 Rubia Species 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- -1 naphthoquinone compounds Chemical class 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 229930192627 Naphthoquinone Natural products 0.000 description 2
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Chemical class CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 description 2
- 239000000539 dimer Chemical class 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 208000034507 Haematemesis Diseases 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 241001107098 Rubiaceae Species 0.000 description 1
- ABRVLXLNVJHDRQ-UHFFFAOYSA-N [2-pyridin-3-yl-6-(trifluoromethyl)pyridin-4-yl]methanamine Chemical compound FC(C1=CC(=CC(=N1)C=1C=NC=CC=1)CN)(F)F ABRVLXLNVJHDRQ-UHFFFAOYSA-N 0.000 description 1
- 150000004056 anthraquinones Chemical class 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 208000001780 epistaxis Diseases 0.000 description 1
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229940124600 folk medicine Drugs 0.000 description 1
- 238000003929 heteronuclear multiple quantum coherence Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 150000002791 naphthoquinones Chemical class 0.000 description 1
- AICOOMRHRUFYCM-ZRRPKQBOSA-N oxazine, 1 Chemical compound C([C@@H]1[C@H](C(C[C@]2(C)[C@@H]([C@H](C)N(C)C)[C@H](O)C[C@]21C)=O)CC1=CC2)C[C@H]1[C@@]1(C)[C@H]2N=C(C(C)C)OC1 AICOOMRHRUFYCM-ZRRPKQBOSA-N 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 230000008736 traumatic injury Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/20—Carbocyclic rings
- C07H15/24—Condensed ring systems having three or more rings
- C07H15/244—Anthraquinone radicals, e.g. sennosides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses 3 compounds in madder and an extraction and separation method, wherein the compounds provided by the invention are named as follows: 2-methyl-1, 6-dihydroxy-9, 10-anthraquinone-3-xylose- (1.fwdarw.2) and preparation method thereofβD-glucose (1), 1,3, 6-trihydroxy-2-hydroxymethyl-9, 10-anthraquinone-3-O‑β-D-glucose (2), 1-hydroxy-2-hydroxymethyl-9, 10-anthraquinone-11-O- β -D-glucose (3). The invention adopts modern spectrum technology such as 1 H NMR、 13 C NMR, two-dimensional nuclear magnetic spectrum, high resolution mass spectrum and physical and chemical properties of the compound are used for carrying out structural identification on the monomer compound obtained by separation, deducing the molecular structure of the compound, and providing a material basis for further quality control and drug effect research of the madder.
Description
Technical Field
The invention relates to the field of traditional Chinese medicine extraction and separation, in particular to a compound extracted, separated and identified from a medicinal material of a hook Mao Qiancao and an extraction and separation method thereof.
Background
The hook Mao Qiancao is Rubiaceae Rubia (Rubiaceae)Rubia) Plant hook Mao Qiancao%Rubia oncotrichaHand-mazz.), is distributed in southwest, northeast to southwest and northwest and southwest of Guizhou, etc., and is collected in "quality standard of Chinese medicinal materials and national medicinal materials of Guizhou" (2003 edition), which is a minority medicine of Guizhou province, and has effects of cooling blood, stopping bleeding, relieving cough and eliminating phlegm, etc., and can be used for treating hematemesis, epistaxis, traumatic injury, etc. Modern researches have found that the main ingredients of the madder are naphthoquinone, anthraquinone, terpenoid, dimer compounds and the like.
At present, the research on the chemical components of the madder is less, only Itawa Hideji group and Tan Ninghua group are used for researching the madder, and the main components are naphthoquinone compounds, anthraquinone compounds, terpenoid compounds and dimer compounds, however, the number of the separated and identified compounds is less.
Disclosure of Invention
The invention combines folk medicine method (water decoction or wine soaking) to perform chemical composition research on the n-butyl alcohol extraction part of 70% ethanol extract of the medicinal material of the hook Mao Qiancao, provides three compounds in the radix Rubiae, and simultaneously provides a simple and rapid extraction and separation method for the compounds of the invention, so as to realize the purpose of enriching the substance components of the medicinal material of the hook Mao Qiancao and providing theoretical basis for research on the later-period pharmacodynamic substance basis.
In order to achieve the above object of the present invention, the present invention provides the following technical solutions:
the compounds provided by the invention are named: 2-methyl-1, 6-dihydroxy-9, 10-anthraquinone-3-xylose- (1.fwdarw.2) and preparation method thereofβD-glucose (1), 1,3, 6-trihydroxy-2-hydroxymethyl-9, 10-anthraquinone-3-O-β-D-glucose (2), 1-hydroxy-2-hydroxymethyl-9, 10-anthraquinone-11-O- β -D-glucose (3), having the structural formula:
/> />。
the invention also provides a method for extracting and separating the compound in the madder, which comprises the following specific steps:
step 1, cutting dry Mao Qiancao roots and rhizomes of the hooks into small sections, reflux-extracting with 70% ethanol for 3 times, extracting with 10 times of volume for 2 h for the first time, extracting with 8 times of volume for 1.5 h for the second time, extracting with 8 times of volume for 1.5 h for the third time, mixing the 3 times of extracting solutions, recovering ethanol under reduced pressure, and volatilizing until no alcohol smell exists to obtain extract.
And 2, dissolving and dispersing the extract with water, and then extracting with petroleum ether, ethyl acetate and n-butanol respectively to obtain a petroleum ether part, an ethyl acetate part, an n-butanol part and a water part respectively.
And 3, taking n-butanol sections, separating by normal phase silica gel column chromatography, gradient eluting by methylene dichloride-methanol (50:1-1:1), detecting by thin layer chromatography, and combining similar components by different color development modes to obtain 7 components Fr.1-Fr.7.
And 4, subjecting Fr.2 to normal phase silica gel column chromatography, eluting with ethyl acetate-methanol (15:1-1:1), and detecting and combining the same components by thin layer chromatography to obtain a plurality of components. Eluting one of the components with ethyl acetate-methanol (8:1) to obtain different sub-components, and subjecting one of the sub-components to normal phase silica gel column chromatography, such as dichloromethane: gradient eluting with methanol (8:1-4:1), and subjecting to Toyopearl HW-40F column chromatography with methanol and chloroform respectively: eluting with methanol (1:1) to obtain compound 1.
And 5, subjecting Fr.4 to Sephadex LH-20 column chromatography, eluting with methanol, and detecting and combining the same fractions by thin layer chromatography to obtain a plurality of components. Subjecting one of the components to Toyopearl HW-40F column chromatography and methanol elution to obtain different subfractions, and subjecting one of the subfractions to MCI column chromatography, methanol-water elution and Toyopearl HW-40F column and methanol elution to obtain compound 2.
And 6, eluting Fr.6 by normal phase silica gel column chromatography and ethyl acetate-methanol (15:1-1:1), detecting and combining the same fractions by thin layer chromatography to obtain a plurality of components. Subjecting one of the components to Sephadex LH-20 column chromatography, chloroform: methanol (1:1) elution yields the different subfractions. Then, a certain subfraction is subjected to Sephadex LH-20 column chromatography and methanol elution to obtain a plurality of small fractions, and then, the fractions are subjected to Sephadex LH-20 column chromatography and chloroform: methanol (1:1) to obtain 2 components, subjecting one component to normal phase silica gel column chromatography, eluting with ethyl acetate-methanol (15:1-1:1), and eluting with Toyopearl HW-40F column chromatography to obtain compound 3.
The invention has the following technical effects: the invention adopts modern spectrum technology such as 1 H NMR、 13 C NMR, two-dimensional nuclear magnetic spectrum, high resolution mass spectrum and physical and chemical properties of the compound are used for carrying out structural identification on the monomer compound obtained by separation, deducing the molecular structure of the compound, and providing a material basis for further quality control and drug effect research of the madder.
Drawings
FIG. 1 HR-ESI-MS of compound 1 of the present invention;
FIG. 2 HR-ESI-MS of compound 2 of the present invention;
FIG. 3 HR-ESI-MS of compound 3 of the present invention;
FIG. 4 Compound 1 of the present invention 1 H-NMR chart (DMSO);
FIG. 5 Compound 1 of the present invention 13 C-NMR chart (DMSO);
FIG. 6 Compound 2 1 H-NMR chart (DMSO);
FIG. 7 Compound 2 1 H-NMR chart (DMSO);
FIG. 8 HMBC pattern (DMSO) of Compound 2;
FIG. 9 Compound 3 1 H-NMR chart (DMSO);
FIG. 10 Compound 3 13 C-NMR chart (DMSO);
FIG. 11 HMBC pattern (DMSO) of Compound 3;
FIG. 12 HMQC map (DMSO) of Compound 3.
Description of the embodiments
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. The technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other. The ingredients or materials involved in the methods described below, unless otherwise specified, are commercially available. The related experimental methods are all conventional methods in the technical field unless specifically described. Wherein the numerical values or numerical proportions, unless noted otherwise, refer to the mass values or mass proportions.
Examples
Step 1: cutting dried root and rhizome of ramulus Uncariae cum Uncis Mao Qiancao and 5 kg into small segments of about 1-2cm, reflux-extracting with 70% ethanol for 3 times, extracting with 10 times of volume for 2 h times, 8 times of volume for 1.5 h times, and 8 times of volume for 1.5 h times, mixing the 3 times of extractive solutions, recovering ethanol under reduced pressure, and volatilizing to obtain extract.
Step 2: the extract is dissolved and dispersed by water, and then is extracted by petroleum ether, ethyl acetate and n-butanol respectively in sequence to obtain petroleum ether part (55 g), ethyl acetate part (65 g), n-butanol part (215 g) and water part (257 g) respectively.
Step 3: separating n-butanol segment by normal phase silica gel column chromatography, gradient eluting with dichloromethane-methanol (50:1-1:1), detecting by thin layer chromatography, and combining similar components to obtain 7 components Fr.1-Fr.7.
Step 4: fr.2 (44.08 g) was eluted with normal phase silica gel column chromatography, ethyl acetate-methanol (15:1-1:1), and the same fractions were combined by thin layer chromatography detection to give 9 fractions (Fr.2.1 to Fr.2.9). Fr.2.4 was subjected to normal phase silica gel column chromatography eluting with ethyl acetate-methanol (8:1) to give 5 fractions (Fr.2.4.1-Fr.2.4.5), wherein Fr.2.4.5 was subjected to normal phase silica gel column chromatography, dichloromethane: gradient eluting with methanol (8:1-4:1), and subjecting to Toyopearl HW-40F column chromatography with methanol and chloroform respectively: methanol (1:1) to give compound 1 (10 mg).
Step 5: fr.4 (10.72 g) was subjected to Sephadex LH-20 column chromatography, methanol elution, and thin layer chromatography to detect the combined same fractions, yielding 6 fractions (Fr.4.1-Fr.4.6). Fr.4.4 was subjected to Toyopearl HW-40F column chromatography and methanol was eluted to obtain 3 fractions (Fr.4.4.1 to Fr.4.4.3), then Fr.4.4.2 was subjected to MCI column chromatography, methanol-water elution, toyopearl HW-40F column and methanol was eluted to obtain compound 2 (9 mg).
Step 6: fr.6 (14.00 g) was subjected to normal phase silica gel column chromatography, ethyl acetate-methanol (15:1-1:1) gradient elution, and the same fractions were combined by thin layer chromatography detection to obtain 5 fractions (Fr.6.1 to Fr.6.5). Fr.6.3 was purified by Sephadex LH-20 column chromatography, chloroform: methanol (1:1) to obtain 6 components (Fr.6.3.1-Fr.6.3.6). Then Fr.6.3.1 is subjected to Sephadex LH-20 column chromatography and methanol is eluted to obtain 6 components (Fr.6.3.1.1-Fr.6.3.1.6), and Fr.6.3.1.4 is subjected to Sephadex LH-20 column chromatography and chloroform: methanol (1:1) is eluted to obtain 2 components (Fr.6.3.1.4.1-Fr.6.3.1.4.2), wherein Fr.6.3.1.4.1 is subjected to normal phase silica gel column chromatography, ethyl acetate-methanol (15:1-1:1) gradient elution is carried out, toyopearl HW-40F column chromatography is carried out, and the methanol is eluted to obtain a compound 3 (8.7 mg).
Conditions for TLC detection performed in the present invention: color developer a: fluorescence was observed under an ultraviolet lamp (254 nm,365 nm); developer b: iodine color development; color developer c:10% sulfuric acid ethanol.
And (3) structural identification: using modern spectroscopic techniques, e.g. 1 H NMR、 13 C NMR, two-dimensional nuclear magnetic spectrum and high-resolution mass spectrum to carry out structural identification on the monomer compound obtained by separation.
Compound 1: pale yellow amorphous powder. HR-ESI-MSm/z: 563.1400 [M–H] – (calculated value: 563.1399), molecular formula C 26 H 28 O 14 The compound was determined by spectroscopic techniques to be 2-methyl-1, 6-dihydroxy-9, 10-anthraquinone-3-xylose- (1→2) - β -D-glucose. The nuclear magnetic data are shown in Table 1.
Compound 2: pale yellow amorphous powder. HR-ESI-MSm/z: 447.0920 [M-H] - (calculated value: 447.0921), molecular formula C 21 H 20 O 11 Determination of the Compound as 1,3, 6-trihydroxy-2-hydroxymethyl-9, 10-anthraquinone-3-by spectroscopic techniquesO-β-D-glucose. The nuclear magnetic data are shown in Table 2.
Compound 3: pale yellow amorphous powder. HR-ESI-MSm/z: 415.1025 [M-H] - (calculated value: 415.1023), molecular formula C 21 H 20 O 9 Determination of the Compound as 1-hydroxy-2-hydroxymethyl-9, 10-anthraquinone-11-by spectroscopic techniquesO-β-D-glucose. The nuclear magnetic data are shown in Table 3.
TABLE 1 Compound 1 1 H-NMR (DMSO, 400 MHz) and 13 C-NMR (DMSO, 100 MHz) data
| Pos. | δ C | δ H |
| 1 | 163.8 | |
| 2 | 120.6 | |
| 3 | 160.4 | |
| 4 | 105.6 | 7.38, 1H, s |
| 4a | 135.3 | |
| 5 | 112.7 | 7.46, 1H, s |
| 6 | 161.3 | |
| 7 | 121.5 | 7.22, 1H, d, J = 8.0 Hz |
| 8 | 129.7 | 8.09, 1H, d, J = 8.8 HZ |
| 8a | 124.3 | |
| 9 | 186.4 | |
| 9a | 110.6 | |
| 10 | 181.7 | |
| 10a | 132.0 | |
| CH 3 | 2.13, 3H, s | |
| 1′ | 97.9 | 4.49, 1H, d, J = 7.6 Hz |
| 2′ | 82.4 | |
| 3′ | 76.2 | |
| 4′ | 68.9 | |
| 5′ | 75.9 | |
| 6′ | 60.2 | |
| 1″ | 105.1 | 5.32, 1H, d, J = 5.6 Hz |
| 2″ | 74.6 | |
| 3″ | 77.1 | |
| 4″ | 69.4 | |
| 5″ | 68.9 |
TABLE 2 Compound 2 1 H-NMR (DMSO, 400 MHz) and 13 C-NMR (DMSO, 100 MHz) data
| Pos. | δ C | δ H | HMBC |
| 1 | 161.6 | ||
| 2 | 123.8 | ||
| 3 | 161.8 | ||
| 4 | 106.1 | 7.41, 1H, s | C-2, 3, 10, 4a, 9a |
| 4a | 133.8 | ||
| 5 | 112.9 | 7.48, 1H, d, J = 2.4 Hz | C-6, 7, 10, 8a |
| 6 | 164.3 | ||
| 7 | 121.7 | 7.24, 1H, dd, J = 2.8, 8.8 Hz | C-5, 8a |
| 8 | 129.7 | 8.09, 1H, d, J = 8.8 HZ | C-6, 9, 10a |
| 8a | 124.1 | ||
| 9 | 186.3 | ||
| 9a | 111.1 | ||
| 10 | 181.7 | ||
| 10a | 135.3 | ||
| 11 | 50.9 | 4.62, 1H, d, J = 11.2 Hz | C-1, 2, |
| 4.54, 1H, d, J = 11.2 Hz | C-1, 2, | ||
| 1′ | 100.9 | 5.06, 1H, d, J = 7.6 Hz | C-3 |
| 2′ | 73.4 | ||
| 3′ | 77.4 | ||
| 4′ | 69.4 | ||
| 5′ | 76.0 | ||
| 6′ | 60.4 |
TABLE 3 Compound 3 1 H-NMR (DMSO, 400 MHz) and 13 C-NMR (DMSO, 100MHz)
| Pos. | δ C | δ H | HMBC |
| 1 | 158.5 | ||
| 2 | 134.1 | ||
| 3 | 134.6 | 8.04, 1H, d, J = 7.6 Hz | C-1, 4a, 11 |
| 4 | 118.6 | 7.74, 1H, d, J = 7.6 Hz | C-2, 9a, 10 |
| 4a | 131.8 | ||
| 5 | 126.9 | 8.19, 1H, m | C-7, 10 |
| 6 | 135.2 | 7.95, 1H, m | C-5, 8, 8a, 10a |
| 7 | 134.7 | 7.95, 1H, m | C-5, 8, 8a, 10a |
| 8 | 126.6 | 8.24, 1H, m | C-6, 9 |
| 8a | 133.2 | ||
| 9 | 188.6 | ||
| 9a | 115.2 | ||
| 10 | 181.8 | ||
| 10a | 132.8 | ||
| 11 | 64.0 | 4.91, 1H, d, J = 15.2 Hz | C-1, 2, 1′ |
| 4.76, 1H, d, J = 15.2 Hz | C-1, 2, 1′ | ||
| 1′ | 102.7 | 4.34, 1H, d, J = 7.6 Hz | C-11 |
| 2′ | 73.6 | ||
| 3′ | 77.1 | ||
| 4′ | 70.0 | ||
| 5′ | 76.7 | ||
| 6′ | 61.0 |
although the present invention has been described with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing examples, or equivalents may be substituted for elements thereof, and any modifications, equivalents, improvements and changes may be made without departing from the spirit and principles of the present invention.
Claims (3)
1. The extraction and separation method of 3 compounds in the madder is characterized by comprising the following steps:
step 1: repeatedly reflux-extracting ramulus Uncariae cum Uncis Mao Qiancao with ethanol, mixing the extractive solutions, and concentrating to obtain extract;
step 2: extracting the extract obtained in the step 1 with petroleum ether, ethyl acetate and n-butanol in sequence to obtain petroleum ether part, ethyl acetate part, n-butanol part and water part respectively;
step 3: taking the n-butanol part obtained in the step 2, performing crude separation by normal phase silica gel column chromatography, detecting by thin layer chromatography, developing, and combining the developed elution parts to obtain 7 components Fr.1-Fr.7;
step 4: subjecting Fr.2 obtained in step 3 to normal phase column chromatography, eluting with ethyl acetate-methanol, and detecting and mixing the same flow components by thin layer chromatography to obtain a plurality of components; wherein, a certain component is separated by normal phase silica gel column chromatography and Toyopearl HW-40F column chromatography to obtain the compound 1;
step 5: subjecting Fr.4 obtained in step 3 to Sephadex LH-20 column chromatography, eluting with methanol, and detecting and mixing the same flow components by thin layer chromatography to obtain a plurality of components; wherein, a certain component is repeatedly eluted by a Toyopearl HW-40F column chromatography, MCI and Toyopearl HW-40F column chromatography to obtain a compound 2;
step 6: subjecting Fr.6 obtained in step 3 to normal phase silica gel column chromatography, eluting with ethyl acetate-methanol, and detecting and mixing the same components by thin layer chromatography to obtain a plurality of components; wherein, a certain component is repeatedly eluted by Sephadex LH-20 column chromatography and normal phase silica gel column chromatography to obtain a compound 3;
extraction to obtain 3 compounds in the madder, wherein the names of the 3 compounds are as follows: 2-methyl-1, 6-dihydroxy-9, 10-anthraquinone-3-xylose- (1.fwdarw.2) and preparation method thereofβD-glucose (1), 1,3, 6-trihydroxy-2-hydroxymethyl-9, 10-anthraquinone-3-O-βD-glucose (2), 1-hydroxy-2-hydroxymethyl-9, 10 anthraquinone-11-O-β-D-glucose (3); the corresponding structural formula is as follows:
。
2. the method for preparing three compounds in the madder root according to claim 1, characterized in that: in step 1, the reflux extraction is performed 3 times with 70% ethanol, wherein the first 10 times of volume extraction is 2. 2 h, the second 8 times of volume extraction is 1.5 h, and the third 8 times of volume extraction is 1.5 h.
3. The method for preparing three compounds in the madder root according to claim 1, characterized in that: in the steps 3,4 and 6, silica gel with 100-300 meshes is selected for chromatographic separation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011078958.4A CN112194690B (en) | 2020-10-10 | 2020-10-10 | 3 compounds in radix Rubiae and extraction and separation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011078958.4A CN112194690B (en) | 2020-10-10 | 2020-10-10 | 3 compounds in radix Rubiae and extraction and separation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112194690A CN112194690A (en) | 2021-01-08 |
| CN112194690B true CN112194690B (en) | 2023-10-20 |
Family
ID=74012860
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011078958.4A Active CN112194690B (en) | 2020-10-10 | 2020-10-10 | 3 compounds in radix Rubiae and extraction and separation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112194690B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1397541A (en) * | 2001-07-20 | 2003-02-19 | 中国人民解放军军事医学科学院放射医学研究所 | Trijuganone and its derivative, and its preparing process and application |
| CN103550237A (en) * | 2013-11-11 | 2014-02-05 | 南京中医药大学 | Composition of active ingredients of rubia cordifolia as well as application of composition in medicines |
-
2020
- 2020-10-10 CN CN202011078958.4A patent/CN112194690B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1397541A (en) * | 2001-07-20 | 2003-02-19 | 中国人民解放军军事医学科学院放射医学研究所 | Trijuganone and its derivative, and its preparing process and application |
| CN103550237A (en) * | 2013-11-11 | 2014-02-05 | 南京中医药大学 | Composition of active ingredients of rubia cordifolia as well as application of composition in medicines |
Non-Patent Citations (2)
| Title |
|---|
| Zhe Wang 等.Rubipodanones A-D, naphthohydroquinone dimers from the roots and rhizomes of Rubia podantha.Phytochemistry.2018,第145卷第153-160页. * |
| 王素贤 等.茜草总蒽醌类成分的研究.药学学报.1992,第27卷(第10期),第743-747页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112194690A (en) | 2021-01-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103739586A (en) | Method for extracting diterpenoid compounds from Blumea aromatic DC. | |
| CN112409422B (en) | Method for extracting ethyl-alpha-D-arabinofuranose from seeds of Nigella sativa | |
| CN112194690B (en) | 3 compounds in radix Rubiae and extraction and separation method | |
| CN105131063B (en) | From Meconopsis integrifolia spend in and meanwhile the method that isolates and purifies a variety of flavones ingredients | |
| CN101845037B (en) | Method for separating xanthione chemical component in Swertia mussoti | |
| CN108047166B (en) | Carpesium nasutum extract and extraction method thereof | |
| CN109232681A (en) | The separation of verbascoside and identification method in a kind of sweet osmanthus | |
| CN112920151B (en) | Isopentene-based flavonoid compound and preparation method and application thereof | |
| CN109053641B (en) | Dineolignan compound, and separation preparation method and application thereof | |
| CN113666894A (en) | Method for extracting and separating furanone compounds from Litsea coreana and application thereof | |
| CN103113439A (en) | Method for preparing kaempferol-3-O-Beta-D-glucuronide in euphorbia sororia | |
| CN105384784A (en) | Screening and separating preparation method for three stilbene polyphenol substances with antioxidant activity in polygonum multiflorum polygonum multiflorumcultivated in Qinghai | |
| CN102603833B (en) | Extraction and separation process of apigenin-7-O-beta-D-glucopyranside from garden balsam stem | |
| CN107043363B (en) | A method of extracting scopoletin from poppyhead stool fruit | |
| CN101318981B (en) | Method for separating benzoxazole oxazinone glycoside compounds from acanthus | |
| CN105085498B (en) | The method and system of extraction separation isovitexin from Desmodium styracifolium | |
| CN113735922B (en) | Method for extracting lignans or terpenoids from cymbidium sinense | |
| CN108586396A (en) | - 2 (5H)-furanones of 3- (4 '-hydroxyls -3 '-methoxy-benzyl) and its extracting method | |
| CN104650053A (en) | Flavonoids compound with novel structure, as well as preparation method and applications thereof | |
| CN109956984B (en) | Method for extracting and separating nicotiflorin from China rose | |
| CN112390833B (en) | Method for separating and extracting sugar and sugar derivatives from Nigellan sativa seeds | |
| CN115304616B (en) | Gastrodia elata active compound and preparation method and application thereof | |
| CN108892698A (en) | A method of utilizing compound in high speed adverse current chromatogram separation Fructus Aurantii | |
| CN110003293B (en) | Method for extracting parasitoside from Chinese rose | |
| CN114805383B (en) | Method for extracting dimer furanone compounds from hawk tea and application of dimer furanone compounds |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |