CN104370814A - Method for extracting glaucine from pachysandra terminalis Sieb.et Zucc. - Google Patents
Method for extracting glaucine from pachysandra terminalis Sieb.et Zucc. Download PDFInfo
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- CN104370814A CN104370814A CN201410280446.4A CN201410280446A CN104370814A CN 104370814 A CN104370814 A CN 104370814A CN 201410280446 A CN201410280446 A CN 201410280446A CN 104370814 A CN104370814 A CN 104370814A
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- glaucine
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- RUZIUYOSRDWYQF-HNNXBMFYSA-N (S)-glaucine Chemical compound CN1CCC2=CC(OC)=C(OC)C3=C2[C@@H]1CC1=C3C=C(OC)C(OC)=C1 RUZIUYOSRDWYQF-HNNXBMFYSA-N 0.000 title claims abstract description 37
- DABPOQZSGVNAAS-UHFFFAOYSA-N Glaucocalactone Natural products O=CC12C3C(C4)OC(=O)C2C(C)(C)CCC1OC(=O)C13CC4C(=C)C1OC(=O)C DABPOQZSGVNAAS-UHFFFAOYSA-N 0.000 title claims abstract description 37
- 229930004041 glaucine Natural products 0.000 title claims abstract description 37
- 229940113086 glaucine Drugs 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 32
- 241001279838 Pachysandra terminalis Species 0.000 title abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 51
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000000741 silica gel Substances 0.000 claims abstract description 32
- 229910002027 silica gel Inorganic materials 0.000 claims abstract description 32
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 claims abstract description 29
- 229930013930 alkaloid Natural products 0.000 claims abstract description 22
- 238000010828 elution Methods 0.000 claims abstract description 17
- 239000002904 solvent Substances 0.000 claims abstract description 17
- 229920005654 Sephadex Polymers 0.000 claims abstract description 6
- 239000000284 extract Substances 0.000 claims description 33
- 229960001866 silicon dioxide Drugs 0.000 claims description 30
- 239000007788 liquid Substances 0.000 claims description 28
- 239000011347 resin Substances 0.000 claims description 26
- 229920005989 resin Polymers 0.000 claims description 26
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 22
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 239000003153 chemical reaction reagent Substances 0.000 claims description 14
- 238000004064 recycling Methods 0.000 claims description 14
- 238000010992 reflux Methods 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 8
- 238000010829 isocratic elution Methods 0.000 claims description 7
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 6
- 230000003213 activating effect Effects 0.000 claims description 6
- 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 6
- 230000003252 repetitive effect Effects 0.000 claims description 6
- 238000005201 scrubbing Methods 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000012507 Sephadex™ Substances 0.000 claims description 5
- 150000001768 cations Chemical class 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- NSOXQYCFHDMMGV-UHFFFAOYSA-N Tetrakis(2-hydroxypropyl)ethylenediamine Chemical compound CC(O)CN(CC(C)O)CCN(CC(C)O)CC(C)O NSOXQYCFHDMMGV-UHFFFAOYSA-N 0.000 claims description 2
- 239000012141 concentrate Substances 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- 238000011068 loading method Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 239000003480 eluent Substances 0.000 abstract description 7
- 239000000126 substance Substances 0.000 abstract description 7
- 238000000605 extraction Methods 0.000 abstract description 4
- 229930014626 natural product Natural products 0.000 abstract description 2
- 238000005406 washing Methods 0.000 abstract 1
- 150000001875 compounds Chemical class 0.000 description 9
- -1 aporphine compound Chemical class 0.000 description 8
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 6
- GPTFURBXHJWNHR-UHFFFAOYSA-N protopine Chemical compound C1=C2C(=O)CC3=CC=C4OCOC4=C3CN(C)CCC2=CC2=C1OCO2 GPTFURBXHJWNHR-UHFFFAOYSA-N 0.000 description 6
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 5
- 150000003797 alkaloid derivatives Chemical group 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000007791 liquid phase Substances 0.000 description 4
- 235000012054 meals Nutrition 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 238000010898 silica gel chromatography Methods 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 238000005481 NMR spectroscopy Methods 0.000 description 3
- GTRPODKMSBFDOI-UHFFFAOYSA-N Protopine Natural products CN1Cc2c3OCOc3ccc2C4C1Cc5cc6OCOc6cc5C4=O GTRPODKMSBFDOI-UHFFFAOYSA-N 0.000 description 3
- ZAALQOFZFANFTF-UHFFFAOYSA-N Pseudoprotipine Natural products C1=C2C(=O)CC3=CC=4OCOC=4C=C3CN(C)CCC2=CC2=C1OCO2 ZAALQOFZFANFTF-UHFFFAOYSA-N 0.000 description 3
- CHWPMFMUQATVNK-ARYYTZDLSA-N dihydrosporogen AO-1 Natural products O[C@H]1[C@]2(C(C)=C)O[C@@H]2[C@]2(C)[C@@H](C)[C@H](O)CCC2=C1 CHWPMFMUQATVNK-ARYYTZDLSA-N 0.000 description 3
- 235000013399 edible fruits Nutrition 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 108090000312 Calcium Channels Proteins 0.000 description 1
- 102000003922 Calcium Channels Human genes 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 206010013954 Dysphoria Diseases 0.000 description 1
- 240000005086 Glaucium flavum Species 0.000 description 1
- 235000000629 Glaucium flavum Nutrition 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 241001279833 Pachysandra Species 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 240000000103 Potentilla erecta Species 0.000 description 1
- 235000016551 Potentilla erecta Nutrition 0.000 description 1
- QCMQEZNBBPGFKQ-UHFFFAOYSA-N Thalisopynine Natural products CN1CCC2=C(OC)C(OC)=C(OC)C3=C2C1CC1=C3C=C(OC)C(O)=C1 QCMQEZNBBPGFKQ-UHFFFAOYSA-N 0.000 description 1
- 206010052428 Wound Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000001466 anti-adreneric effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- BZKUYNBAFQJRDM-UHFFFAOYSA-N aporphine Natural products C12=CC=CC=C2CC2N(C)CCC3=CC=CC1=C32 BZKUYNBAFQJRDM-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940124600 folk medicine Drugs 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000006317 isomerization reaction Methods 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 150000002596 lactones Chemical class 0.000 description 1
- FVRABHGHBLRNNR-UHFFFAOYSA-N liriodenine Natural products O=C1C=CC=c2c1cc3nccc4cc5OCOc5c2c34 FVRABHGHBLRNNR-UHFFFAOYSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 208000007106 menorrhagia Diseases 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000005311 nuclear magnetism Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000019633 pungent taste Nutrition 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 230000011218 segmentation Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 125000003039 tetrahydroisoquinolinyl group Chemical class C1(NCCC2=CC=CC=C12)* 0.000 description 1
- 229940075420 xanthine Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
- C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
- C07D221/04—Ortho- or peri-condensed ring systems
- C07D221/18—Ring systems of four or more rings
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a method for extracting glaucine from pachysandra terminalis Sieb.et Zucc., and belongs to the technical field of extraction of natural products. The method comprises the following steps: 1) getting a pachysandra terminalis Sieb.et Zucc. total alkaloids extractum, applying to an alkaline silica gel column, taking a chloroform-methanol system as an eluent for gradient elution, merging fractions and distilling off the solvent, so as to obtain a first column-chromatograph substance; 2) applying the first column-chromatograph substance to a silica gel column, taking a chloroform-methanol system as an eluent for gradient elution, merging fractions with a same main spot, and distilling off the solvent, so as to obtain a second column-chromatograph substance; 3) applying the second column-chromatograph substance to a dextran gel column, taking a chloroform-methanol system as an eluent for gradient elution, merging fractions with a same main spot, and distilling off the solvent, so as to obtain a third column-chromatograph substance; and 4) repeatedly washing the third column-chromatograph substance with methanol, so as to obtain glaucine. The method is simple in operation, low in cost and high in extraction efficiency, and the obtained glaucine has the purity of 99% or more.
Description
Technical field
The invention belongs to technical field of natural product extraction, be specifically related to a kind of method extracting glaucine from the celestial rope of bundle.
Background technology
The celestial rope of Folk medicine bundle is the herb of Buxaceae Pachysandra plant poppyhead stool fruit (Pachysandraterminalis Sieb.et Zucc.).Poppyhead stool fruit has another name called top stamen Herba Pachysandrae Stylosae, grass green for a long time, bundle celestial being seven, convulsion grass, Pachysandra terminalis.This plant is alkaloid because of its main component, has effect of remove rheumatism, promoting blood circulation and stopping pain, therefore among the people being mainly used in Shaanxi treats rheumatism paralysis, numb limbs and tense tendons, wound, menorrhagia, dysphoria etc.Up to now, both at home and abroad 60-80 age in the last century Japan thorough scholar of Kikuchi and China Duan Hongquan are only limitted to the research of the celestial rope of bundle and teach report about poppyhead stool fruit tetrahydroisoquinoline alkaloid chemical composition, have no the report of other structure type alkaloid extraction and separation method of this plant.
Bibliographical information glaucine is present in bloodroot horned poppy and Yanhusuo, is aporphine compound, has another name called extra large small-mouthed jar blue.The structural formula of glaucine is:
Modern pharmacology research report, glaucine not only has calcium channel retardation thus produces the physiological effect based on relexation, and antioxygenation can be played by Scavenger of ROS and the mechanism such as living radical, suppression xanthoglobulin-xanthine oxidase system, also there is vitro inhibition cell proliferation and suppress transfer, anti-inflammatory, analgesia, cough-relieving, antithrombus formation, the effects such as antiadrenergic drug.This has good application prospect clinically.
Summary of the invention
The object of the present invention is to provide a kind of method extracting glaucine from the celestial rope of bundle, the method is with low cost, simple to operate, extracts the glaucine purity obtained high.
The present invention is achieved through the following technical solutions:
From the celestial rope of bundle, extract a method for glaucine, comprise the following steps:
1) bundle celestial rope total alkaloids medicinal extract is got, be splined on alkaline silica gel post, with chloroform-methanol system for elutriant, by chloroform: the volume ratio of methyl alcohol=(200:1) ~ (0:1) carries out gradient elution, effluent liquid is detected, by chloroform: the cut of the volume ratio wash-out of methyl alcohol=(25:1) ~ (10:1) merges, solvent evaporated, obtain crossing post part for the first time;
2) first time post part is excessively got, be splined on silicagel column, with chloroform-methanol system for elutriant, by chloroform: the volume ratio of methyl alcohol=(100:1) ~ (2:1) carries out gradient elution, effluent liquid is detected, after merging the identical cut of principal spot, solvent evaporated, obtains second time and crosses post part;
Wherein, the quadrol of 1% is amassed in described chloroform-methanol system elutions liquid containing chloroform-methanol system elutions liquid;
3) second time is crossed post part, be splined on sephadex column, with chloroform-methanol system for elutriant, 1:2 carries out isocratic elution by volume, detects effluent liquid, and after merging the identical cut of principal spot, solvent evaporated, obtains crossing post part for the third time;
4) third time is crossed post part with after methyl alcohol repetitive scrubbing, obtain glaucine.
Step 1) alkaline silica gel that loads in described alkaline silica gel post is in 1g:(2 ~ 5) ratio of mL, the NaOH solution of 0.1 ~ 0.3mol/L is added in silica gel, leave standstill after stirring, incline supernatant liquor, obtains after activating 5 ~ 8h at 100 ~ 120 DEG C.The granularity of used silica gel is 200 ~ 300 orders.
Described to effluent liquid carry out detection be adopt tlc effluent liquid is detected, develop the color with improvement Dragendorff's reagent.
Described sephadex column is Sephadex LH-20 post.
The preparation of described bundle celestial being rope total alkaloids medicinal extract, specifically comprises the following steps:
(1) powder massfraction of bundle celestial being being restricted is the alcohol reflux of 95%, concentrates and obtain medicinal extract after reclaiming extracting solution;
(2) be filter the mass concentration of medicinal extract bundle celestial rope powder quality 8 ~ 12 times of volumes after the dissolving with hydrochloric acid of 1 ~ 2%, by the filtrate that obtains by Zeo-karb, be washed to till effluent liquid is neutrality;
(3) then take out resin cation (R.C.), soak 10 ~ 14 hours with the ammoniacal liquor that the mass concentration of resin cation (R.C.) quality 3 ~ 5 times of volumes is 8 ~ 15%, obtain the resin that alkalizes;
(4) mass fraction adding alkalization resin quality 6 ~ 8 times of volumes in alkalization resin is 95% alcohol reflux 1 ~ 2 hour, recycling design, obtains tying celestial rope total alkaloids medicinal extract.
Described in step (1) is 2 ~ 4 times with the alcohol reflux number of times that massfraction is 95%, each 2 ~ 3h.
Described Zeo-karb is LS-12 strongly-acid macroporous type vinylbenzene Zeo-karb.
The filtrate obtained is 0.5 ~ 1.5BV/h by the loading speed of Zeo-karb by step (2).
Compared with prior art, the present invention has following useful technique effect:
The present invention extracts the method for glaucine from the celestial rope of bundle, be separated first from the celestial rope of Chinese medicinal materials bundle and obtain monomeric compound glaucine, the first parlkaline silicagel column of bundle celestial being rope total alkaloids medicinal extract is carried out first time gradient elution by present method, the identical cut collected is again through the gradient elution of silicagel column repeatedly, finally by sephadex column isocratic elution, be separated and obtain glaucine.The inventive method is simple, with low cost, and the purity being separated the glaucine obtained reaches more than 99%, compared with the alumina column adopted with traditional isolating alkaloids, easier on alkaline silica gel post dress post, bed board inspection identifying operation.The silicagel column adopted with current isolating alkaloids and add a certain amount of alkaline matter (ammoniacal liquor in elutriant, diethylamine, triethylamine etc.) separation method compare, alkaline silica gel post not only avoid pungency organic solvent (triethylamine in the process of initial a large amount of rough segmentation, ammoniacal liquor etc.) a large amount of uses, and saved cost.First time crosses post part and again carries out silicagel column when being separated, and add 1% diethylamine in moving phase, this is not only conducive to alkaline alkaloidal separation, and avoids the decomposition of compound separation and the isomerization of the types such as aldehyde, ketone, lactone, and separating effect is better.
Accompanying drawing explanation
Fig. 1 is that the present invention is separated the protopine obtained
1h-NMR collection of illustrative plates;
Fig. 2 is that the present invention is separated the protopine obtained
13c-NMR collection of illustrative plates;
Embodiment
Below in conjunction with specific embodiment, the present invention is described in further detail, and the explanation of the invention is not limited.
Embodiment 1
From the celestial rope of bundle, extract the method for glaucine, comprise the following steps:
1) bundle celestial being rope meal is about 7.5kg, with 95% alcohol reflux 3 times, each 2h, reclaim extracting solution, concentrating under reduced pressure obtains medicinal extract, by 1.0kg medicinal extract 10L, mass concentration is filter after the dissolving with hydrochloric acid of 2%, by filtrate according to the flow velocity of 1BV/h by after LS-12 strongly-acid macroporous type vinylbenzene Zeo-karb, be washed to till effluent liquid is neutrality.Take out resin resin quality 4 times of volumes, mass concentration be 10% ammoniacal liquor soak 12 hours, in the alkalization resin obtained, adding 95% alcohol reflux 1.5 hours of resin quality 8 times of volumes, recycling design, obtains tying celestial rope total alkaloids medicinal extract 61.8g.
2) get 50.0g bundle celestial rope total alkaloids medicinal extract and cross normal pressure alkaline silica gel column chromatography, with chloroform-methanol system for elutriant, the gradient being (200:1) ~ (0:1) by chloroform and methyl alcohol volume ratio carries out wash-out, equivalent collects elutriant, every part of 500mL, the cut TLC collected detects, with the colour developing of improvement Dragendorff's reagent, merge identical cut, solvent evaporated, obtain 30 parts altogether, called after A1-A30 respectively, collect the cut that elutriant gradient is chloroform-methanol (25:1) ~ (10:1), the i.e. cut of A13 ~ A15, recycling design, evaporate to dryness, obtain crossing post part for the first time.Wherein, described alkaline silica gel is add the NaOH solution that 300mL mass concentration is 0.2mol/L in 200 ~ 300 object silica gel to 100g granularity, and leave standstill after stirring, incline supernatant liquor, obtains after activating 5h at 110 DEG C.
3) first time is crossed post part and cross silicagel column, with chloroform-methanol (1% diethylamine) system for elutriant, (100:1) ~ (2:1) carries out gradient elution by volume, effluent liquid TLC is detected, with the colour developing of improvement Dragendorff's reagent, merge the cut that principal spot is identical, recycling design, evaporate to dryness, obtains second time and crosses post part;
4) second time is crossed post part after Sephadex LH-20 post, with chloroform-methanol system for eluent, 1:2 isocratic elution by volume, detects effluent liquid TLC, with the colour developing of improvement Dragendorff's reagent, merge principal spot cut, solvent evaporated, obtains crossing post part for the third time, after methyl alcohol repetitive scrubbing, obtain monomeric compound, by high-efficient liquid phase chromatogram technique analysis purity more than 99%.
Adopt the methods such as 1H-NMR, 13C-NMR to carry out Structural Identification by being separated the monomeric compound obtained, and in conjunction with bibliographical information, determine that this compound structure is glaucine.
Nuclear magnetic resonance map is with reference to accompanying drawing 1 ~ 2, and nuclear magnetic data is as follows: yellow needles (chloroform), molecular formula C
21h
25nO
4, relative molecular mass 355.
1H-NMR(400MHz,CDCl
3)δppm:8.09(s,1H),6.78(s,1H),6.59(s,1H),3.93(s,3H),3.90(s,3H),3.89(s,3H),3.65(s,3H),3.17(t,J＝14.2Hz,1H),3.03(m,3H),2.73–2.60(m,2H),2.57(s,3H),2.00(m,1H).
13C-NMR(100MHz,CDCl
3)δppm:152.00(C-2),148.00(C-9),147.48(C-10),144.31(C-1),129.19(C-7a),128.77(C-3a),126.93(C-3b,11b),124.45(C-11a),111.58(C-11),110.80(C-8),110.35(C-3),62.50(C-6a),60.20(1-OCH3),55.93(2-OCH3),55.80(9-OCH3),55.78(10-OCH3),53.21(1-OCH3),43.82(N-CH3),34.39(C-7),29.05(C-4)。
The C modal data reported in protopine the present invention obtained and reference is compared, and result is as shown in table 1: (wherein, document 1: Feng waits quietly. the research [J] of alkaloid component in Yanhusuo. Beijing: Chinese experimental pharmacology of traditional Chinese medical formulae magazine, 2013; Document 2: Xu Xianghong etc. the research [J] of alkaloid component in Yanhusuo. Nanjing: China Medicine University's journal, 2002)
The comparison of table 1 glaucine nuclear-magnetism C modal data
As can be seen from Table 1, the present invention is basically identical from tying the data being separated the C modal data of the monomeric compound obtained and the glaucine of bibliographical information celestial rope.
Embodiment 2
From the celestial rope of bundle, extract the method for glaucine, comprise the following steps:
1) bundle celestial being rope meal is about 7.5kg, with 95% alcohol reflux 2 times, each 3h, reclaim extracting solution, concentrating under reduced pressure obtains medicinal extract, by 1.0kg medicinal extract 8L, mass concentration is filter after the dissolving with hydrochloric acid of 1.5%, by filtrate according to the flow velocity of 0.5BV/h by after LS-12 strongly-acid macroporous type vinylbenzene Zeo-karb, be washed to till effluent liquid is neutrality.Take out resin resin quality 3 times of volumes, mass concentration be 15% ammoniacal liquor soak 10 hours, in the alkalization resin obtained, adding 95% alcohol reflux 1.5 hours of resin quality 8 times of volumes, recycling design, obtains tying celestial rope total alkaloids medicinal extract 60.9g.
2) get 50.0g bundle celestial rope total alkaloids medicinal extract and cross normal pressure alkaline silica gel column chromatography, with chloroform-methanol system for elutriant, the gradient being (200:1) ~ (0:1) by chloroform and methyl alcohol volume ratio carries out wash-out, equivalent collects elutriant, every part of 500mL, the cut TLC collected detects, with the colour developing of improvement Dragendorff's reagent, merge identical cut, solvent evaporated, obtain 30 parts altogether, called after A1-A30 respectively, collect the cut that elutriant gradient is chloroform-methanol (25:1) ~ (10:1), the i.e. cut of A13 ~ A15, recycling design, evaporate to dryness, obtain crossing post part for the first time.Wherein, described alkaline silica gel is add the NaOH solution that 3000ml mass concentration is 0.2mol/L in 200 ~ 300 object silica gel to 100g granularity, and leave standstill after stirring, incline supernatant liquor, obtains after activating 5h at 110 DEG C.
3) first time is crossed post part and cross silicagel column, with chloroform-methanol (1% diethylamine) system for elutriant, (1000:1) ~ (2:1) carries out gradient elution by volume, effluent liquid TLC is detected, with the colour developing of improvement Dragendorff's reagent, merge the cut that principal spot is identical, recycling design, evaporate to dryness, obtains second time and crosses post part;
4) second time is crossed post part after Sephadex LH-20 post, with chloroform-methanol system for eluent, 1:2 isocratic elution by volume, detects effluent liquid TLC, with the colour developing of improvement Dragendorff's reagent, merge principal spot cut, solvent evaporated, obtain crossing post part for the third time, after methyl alcohol repetitive scrubbing, obtain monomeric compound, i.e. glaucine, by high-efficient liquid phase chromatogram technique analysis purity more than 99%.
Embodiment 3
From the celestial rope of bundle, extract the method for glaucine, comprise the following steps:
1) bundle celestial being rope meal is about 7.5kg, with 95% alcohol reflux 4 times, each 2h, reclaim extracting solution, concentrating under reduced pressure obtains medicinal extract, by 1.0kg medicinal extract 12L, mass concentration is filter after the dissolving with hydrochloric acid of 1%, by filtrate according to the flow velocity of 1.5BV/h by after LS-12 strongly-acid macroporous type vinylbenzene Zeo-karb, be washed to till effluent liquid is neutrality.Take out resin resin quality 4 times of volumes, mass concentration be 8% ammoniacal liquor soak 14 hours, in the alkalization resin obtained, adding 95% alcohol reflux 1.5 hours of resin quality 8 times of volumes, recycling design, obtains tying celestial rope total alkaloids medicinal extract 60.2g.
2) get 50.0g bundle celestial rope total alkaloids medicinal extract and cross normal pressure alkaline silica gel column chromatography, with chloroform-methanol system for elutriant, the gradient being (200:1) ~ (0:1) by chloroform and methyl alcohol volume ratio carries out wash-out, equivalent collects elutriant, every part of 500mL, the cut TLC collected detects, with the colour developing of improvement Dragendorff's reagent, merge identical cut, solvent evaporated, obtain 30 parts altogether, called after A1-A30 respectively, collect the cut that elutriant gradient is chloroform-methanol (25:1) ~ (10:1), the i.e. cut of A13 ~ A15, recycling design, evaporate to dryness, obtain crossing post part for the first time.Wherein, described alkaline silica gel is add the NaOH solution that 300ml mass concentration is 0.2mol/L in 200 ~ 300 object silica gel to 100g granularity, and leave standstill after stirring, incline supernatant liquor, obtains after activating 5h at 110 DEG C.
3) first time is crossed post part and cross silicagel column, with chloroform-methanol (1% diethylamine) system for elutriant, (50:1) ~ (10:1) carries out gradient elution by volume, effluent liquid TLC is detected, with the colour developing of improvement Dragendorff's reagent, merge the cut that principal spot is identical, recycling design, evaporate to dryness, obtains second time and crosses post part;
4) second time is crossed post part after Sephadex LH-20 post, with chloroform-methanol system for eluent, 1:2 isocratic elution by volume, detects effluent liquid TLC, with the colour developing of improvement Dragendorff's reagent, merge principal spot cut, solvent evaporated, obtain crossing post part for the third time, after methyl alcohol repetitive scrubbing, obtain monomeric compound, i.e. glaucine, by high-efficient liquid phase chromatogram technique analysis purity more than 99%.
Embodiment 4
From the celestial rope of bundle, extract the method for glaucine, comprise the following steps:
1) bundle celestial being rope meal is about 7.5kg, with 95% alcohol reflux 3 times, each 2h, reclaim extracting solution, concentrating under reduced pressure obtains medicinal extract, by 1.0kg medicinal extract 12L, mass concentration is filter after the dissolving with hydrochloric acid of 1%, by filtrate according to the flow velocity of 0.8BV/h by after LS-12 strongly-acid macroporous type vinylbenzene Zeo-karb, be washed to till effluent liquid is neutrality.Take out resin resin quality 4 times of volumes, mass concentration be 8% ammoniacal liquor soak 12 hours, in the alkalization resin obtained, adding 95% alcohol reflux 1.5 hours of resin quality 8 times of volumes, recycling design, obtains tying celestial rope total alkaloids 59.9g.
2) get 50.0g total alkaloids and cross normal pressure alkaline silica gel column chromatography, with chloroform-methanol system for elutriant, the gradient being (200:1) ~ (0:1) by chloroform and methyl alcohol volume ratio carries out wash-out, equivalent collects elutriant, every part of 500mL, the cut TLC collected detects, with the colour developing of improvement Dragendorff's reagent, merge identical cut, solvent evaporated, obtain 30 parts altogether, called after A1-A30 respectively, collect the cut that elutriant gradient is chloroform-methanol (25:1) ~ (10:1), the i.e. cut of A13 ~ A15, recycling design, evaporate to dryness, obtain crossing post part for the first time.Wherein, described alkaline silica gel is add the NaOH solution that 300ml mass concentration is 0.2mol/L in 200 ~ 300 object silica gel to 100g granularity, and leave standstill after stirring, incline supernatant liquor, obtains after activating 5h at 110 DEG C.
3) first time is crossed post part and cross silicagel column, with chloroform-methanol (1% diethylamine) system for elutriant, (100:1) ~ (2:1) carries out gradient elution by volume, effluent liquid TLC is detected, with the colour developing of improvement Dragendorff's reagent, merge the cut that principal spot is identical, recycling design, evaporate to dryness, obtains second time and crosses post part;
4) second time is crossed post part after Sephadex LH-20 post, with chloroform-methanol system for eluent, 1:2 isocratic elution by volume, detects effluent liquid TLC, with the colour developing of improvement Dragendorff's reagent, merge principal spot cut, solvent evaporated, obtain crossing post part for the third time, after methyl alcohol repetitive scrubbing, obtain monomeric compound, i.e. glaucine, by high-efficient liquid phase chromatogram technique analysis purity more than 99%.
Claims (9)
1. from the celestial rope of bundle, extract a method for glaucine, it is characterized in that, comprise the following steps:
1) bundle celestial rope total alkaloids medicinal extract is got, be splined on alkaline silica gel post, with chloroform-methanol system for elutriant, by chloroform: the volume ratio of methyl alcohol=(200:1) ~ (0:1) carries out gradient elution, effluent liquid is detected, by chloroform: the cut of the volume ratio wash-out of methyl alcohol=(25:1) ~ (10:1) merges, solvent evaporated, obtain crossing post part for the first time;
2) first time post part is excessively got, be splined on silicagel column, with chloroform-methanol system for elutriant, by chloroform: the volume ratio of methyl alcohol=(100:1) ~ (2:1) carries out gradient elution, effluent liquid is detected, after merging the identical cut of principal spot, solvent evaporated, obtains second time and crosses post part; Wherein, the quadrol of 1% is amassed in described chloroform-methanol system elutions liquid containing chloroform-methanol system elutions liquid;
3) second time is crossed post part, be splined on sephadex column, with chloroform-methanol system for elutriant, 1:2 carries out isocratic elution by volume, detects effluent liquid, and after merging the identical cut of principal spot, solvent evaporated, obtains crossing post part for the third time;
4) third time is crossed post part with after methyl alcohol repetitive scrubbing, obtain glaucine.
2. a kind of method extracting glaucine from the celestial rope of bundle according to claim 1, it is characterized in that, step 1) alkaline silica gel that loads in described alkaline silica gel post is in 1g:(2 ~ 5) ratio of mL, the NaOH solution of 0.1 ~ 0.3mol/L is added in silica gel, leave standstill after stirring, incline supernatant liquor, obtains after activating 5 ~ 8h at 100 ~ 120 DEG C.
3. a kind of method extracting glaucine from the celestial rope of bundle according to claim 2, it is characterized in that, the granularity of used silica gel is 200 ~ 300 orders.
4. a kind of method extracting glaucine from the celestial rope of bundle according to claim 1, is characterized in that, described to carry out detection to effluent liquid be adopt tlc to detect effluent liquid, develops the color with improvement Dragendorff's reagent.
5. a kind of method extracting glaucine from the celestial rope of bundle according to claim 1, it is characterized in that, described sephadex column is Sephadex LH-20 post.
6. a kind of method extracting glaucine from the celestial rope of bundle according to claim 1, is characterized in that, the preparation of described bundle celestial being rope total alkaloids medicinal extract, specifically comprises the following steps:
(1) powder massfraction of bundle celestial being being restricted is the alcohol reflux of 95%, concentrates and obtain medicinal extract after reclaiming extracting solution;
(2) be filter the mass concentration of medicinal extract bundle celestial rope powder quality 8 ~ 12 times of volumes after the dissolving with hydrochloric acid of 1 ~ 2%, by the filtrate that obtains by Zeo-karb, be washed to till effluent liquid is neutrality;
(3) then take out resin cation (R.C.), soak 10 ~ 14 hours with the ammoniacal liquor that the mass concentration of resin cation (R.C.) quality 3 ~ 5 times of volumes is 8 ~ 15%, obtain the resin that alkalizes;
(4) mass fraction adding alkalization resin quality 6 ~ 8 times of volumes in alkalization resin is 95% alcohol reflux 1 ~ 2 hour, recycling design, obtains tying celestial rope total alkaloids medicinal extract.
7. a kind of method extracting glaucine from the celestial rope of bundle according to claim 6, it is characterized in that, described in step (1) is 2 ~ 4 times with the alcohol reflux number of times that massfraction is 95%, each 2 ~ 3h.
8. a kind of method extracting glaucine from the celestial rope of bundle according to claim 6, it is characterized in that, described Zeo-karb is LS-12 strongly-acid macroporous type vinylbenzene Zeo-karb.
9. a kind of method extracting glaucine from the celestial rope of bundle according to claim 6, is characterized in that, the filtrate obtained is 0.5 ~ 1.5BV/h by the loading speed of Zeo-karb by step (2).
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