CN102432419A - Method for extracting and separating beta-elemene from Eupatorium adenophorum - Google Patents
Method for extracting and separating beta-elemene from Eupatorium adenophorum Download PDFInfo
- Publication number
- CN102432419A CN102432419A CN2011104575235A CN201110457523A CN102432419A CN 102432419 A CN102432419 A CN 102432419A CN 2011104575235 A CN2011104575235 A CN 2011104575235A CN 201110457523 A CN201110457523 A CN 201110457523A CN 102432419 A CN102432419 A CN 102432419A
- Authority
- CN
- China
- Prior art keywords
- elemene
- beta
- volatile oil
- extraction
- solvent
- 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
- OPFTUNCRGUEPRZ-QLFBSQMISA-N (-)-beta-elemene Chemical compound CC(=C)[C@@H]1CC[C@@](C)(C=C)[C@H](C(C)=C)C1 OPFTUNCRGUEPRZ-QLFBSQMISA-N 0.000 title claims abstract description 230
- OPFTUNCRGUEPRZ-UHFFFAOYSA-N (+)-beta-Elemen Natural products CC(=C)C1CCC(C)(C=C)C(C(C)=C)C1 OPFTUNCRGUEPRZ-UHFFFAOYSA-N 0.000 title claims abstract description 115
- 238000000034 method Methods 0.000 title claims abstract description 67
- 244000062748 Eupatorium adenophorum Species 0.000 title abstract description 5
- 238000000605 extraction Methods 0.000 claims abstract description 103
- 239000000341 volatile oil Substances 0.000 claims abstract description 92
- 239000002994 raw material Substances 0.000 claims abstract description 61
- 239000002904 solvent Substances 0.000 claims abstract description 49
- 238000004440 column chromatography Methods 0.000 claims abstract description 20
- 238000002360 preparation method Methods 0.000 claims abstract description 9
- 238000007670 refining Methods 0.000 claims abstract description 5
- 238000000199 molecular distillation Methods 0.000 claims abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 167
- 244000025254 Cannabis sativa Species 0.000 claims description 53
- 241000735527 Eupatorium Species 0.000 claims description 53
- 235000012766 Cannabis sativa ssp. sativa var. sativa Nutrition 0.000 claims description 51
- 235000012765 Cannabis sativa ssp. sativa var. spontanea Nutrition 0.000 claims description 51
- 235000009120 camo Nutrition 0.000 claims description 51
- 235000005607 chanvre indien Nutrition 0.000 claims description 51
- 239000011487 hemp Substances 0.000 claims description 51
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 48
- 239000000284 extract Substances 0.000 claims description 42
- 238000009835 boiling Methods 0.000 claims description 38
- 239000003921 oil Substances 0.000 claims description 38
- 239000012074 organic phase Substances 0.000 claims description 36
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 35
- 239000003960 organic solvent Substances 0.000 claims description 29
- 238000000926 separation method Methods 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 26
- 238000004821 distillation Methods 0.000 claims description 21
- 239000003495 polar organic solvent Substances 0.000 claims description 20
- 238000012545 processing Methods 0.000 claims description 18
- 238000000526 short-path distillation Methods 0.000 claims description 13
- 239000007791 liquid phase Substances 0.000 claims description 9
- 229920005989 resin Polymers 0.000 claims description 9
- 239000011347 resin Substances 0.000 claims description 9
- 238000004587 chromatography analysis Methods 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 8
- 238000000746 purification Methods 0.000 claims description 7
- 238000010898 silica gel chromatography Methods 0.000 claims description 6
- 238000005554 pickling Methods 0.000 claims description 5
- 238000002953 preparative HPLC Methods 0.000 claims description 5
- 238000000194 supercritical-fluid extraction Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
- 238000000874 microwave-assisted extraction Methods 0.000 claims description 4
- 238000007127 saponification reaction Methods 0.000 claims description 4
- 229910052709 silver Inorganic materials 0.000 claims description 4
- 239000004332 silver Substances 0.000 claims description 4
- 238000000638 solvent extraction Methods 0.000 claims description 4
- 238000005187 foaming Methods 0.000 claims description 3
- -1 silver ions Chemical class 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 238000003815 supercritical carbon dioxide extraction Methods 0.000 claims 1
- 241000196324 Embryophyta Species 0.000 abstract description 7
- 239000003480 eluent Substances 0.000 abstract description 7
- 230000003211 malignant effect Effects 0.000 abstract description 4
- 239000012043 crude product Substances 0.000 abstract 2
- 238000004128 high performance liquid chromatography Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 description 50
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 48
- 239000003208 petroleum Substances 0.000 description 20
- 239000007788 liquid Substances 0.000 description 19
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 15
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 12
- 229910021536 Zeolite Inorganic materials 0.000 description 10
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 description 10
- 239000003814 drug Substances 0.000 description 10
- 239000010457 zeolite Substances 0.000 description 10
- 238000005194 fractionation Methods 0.000 description 8
- 101100313763 Arabidopsis thaliana TIM22-2 gene Proteins 0.000 description 6
- 238000010438 heat treatment Methods 0.000 description 6
- 240000007311 Commiphora myrrha Species 0.000 description 5
- 235000006965 Commiphora myrrha Nutrition 0.000 description 5
- 240000009138 Curcuma zedoaria Species 0.000 description 5
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 5
- 240000004784 Cymbopogon citratus Species 0.000 description 5
- 235000017897 Cymbopogon citratus Nutrition 0.000 description 5
- 235000007265 Myrrhis odorata Nutrition 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 238000005070 sampling Methods 0.000 description 5
- 238000001256 steam distillation Methods 0.000 description 5
- 230000009514 concussion Effects 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 238000003811 acetone extraction Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 238000005457 optimization Methods 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 238000013459 approach Methods 0.000 description 2
- 239000012501 chromatography medium Substances 0.000 description 2
- 238000010835 comparative analysis Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 206010065553 Bone marrow failure Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
- 241001513369 Triosteum perfoliatum Species 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000002389 essential drug Substances 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
Landscapes
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
本发明属于植物提取领域，目的是提供一种利用恶性杂草植物紫茎泽兰为原料，从中提取分离β-榄香烯的方法。本发明方法如下：A、原料准备：以紫茎泽兰的茎、叶为原料，鲜品打浆或干燥粉碎；B、挥发油提取：从原料中得到挥发油；C、柱层析：将步骤B得到的挥发油进行柱层析，收集洗脱液，除去洗脱液中的溶剂，得到β-榄香烯粗品；D、精制：将步骤B得到的挥发油或步骤C得到的粗品，采用分子蒸馏或制备型高效液相色谱系统进行精制获得高纯度的β-榄香烯。本发明提取所得β-榄香烯产品纯度高，产率高，成本低，为紫茎泽兰开发利用和治理以及获得β-榄香烯提供一种全新的、廉价的、有效的途径。The invention belongs to the field of plant extraction, and aims to provide a method for extracting and separating β-elemene from the malignant weed plant Eupatorium adenophorum as a raw material. The method of the present invention is as follows: A, raw material preparation: take the stems and leaves of Eupatorium adenophorum as raw materials, fresh products are beaten or dried and pulverized; B, volatile oil extraction: obtain volatile oil from raw materials; C, column chromatography: obtain step B The volatile oil obtained in step B is subjected to column chromatography, the eluent is collected, and the solvent in the eluent is removed to obtain the crude product of β-elemene; D, refining: the volatile oil obtained in step B or the crude product obtained in step C is subjected to molecular distillation or preparation High-purity β-elemene was purified by a high-performance liquid chromatography system. The extracted β-elemene obtained by the invention has high purity, high yield and low cost, and provides a new, cheap and effective way for the development, utilization and treatment of Eupatorium adenophorum and the acquisition of β-elemene.
Description
Technical field
The invention belongs to field of plant extraction, be specifically related to the method for extraction separation beta-elemene from malignant weed plant Hemp Eupatorium.
Background technology
Hemp Eupatorium (Eupatorium adenophorum Spreng), another name are destroyed grass, liberation grass, Herba Eupatorii Odorati etc., are the composite family feverwort.This plant distributes extensively on Yunnan Province of China, Guangxi, Sichuan and other places, and resource is very abundant, belongs to the torrid zone, subtropics invasive weed plant, causes very big harm for local husbandry production, and local ecological security is caused very big threat.
Beta-elemene, Latin is called β-Elemenum, English β-elemene by name, chemistry (1S, 2S, 4R) 1-methyl isophthalic acid-vinyl-2,4-diisopropenyl hexanaphthene by name.Structural formula is following:
Beta-elemene is that China passes through a series of State Commission for Restructuring the Economic Systems projects such as " the Seventh Five-Year Plan ", " eight or five ", " 95 "; It is the anti-cancer agent that independent research goes out from traditional Chinese medicine; Be applied to clinically, and be put into national essential drugs and national medical insurance medication catalogue.Clinical application shows; Beta-elemene not only has very strong killing tumour cell and suppress the tumor growth effect; Can also make body obtain tangible immunoprotection effect through activating immune system; Have characteristics such as anticancer spectrum is wide, efficient, no hepatorenal damage, no bone marrow depression, few side effects, be the mankind from now on one period the internal therapy tumour choice drug, the domestic and international market demand is big.At present, beta-elemene mainly depends on from Chinese medicine curcuma zedoary, myrrh, lemongrass, Herba Solidaginis and extracts, and finds to have from Hemp Eupatorium extraction separation to obtain the report of beta-elemene as yet.Because Chinese medicine curcuma zedoary, myrrh, lemongrass, Herba Solidaginis are Chinese medicine, cost is higher relatively, and wherein the content of beta-elemene is not high, and the cost that extracts purifying is higher, causes the price of beta-elemene very expensive, has seriously hindered applying of this medicine.
Contriver of the present invention finds in the Hemp Eupatorium beta-elemene content far above Chinese medicine curcuma zedoary, myrrh, lemongrass, Herba Solidaginis, and Hemp Eupatorium is the invasive weed plant, and aboundresources, its raw materials cost are also low more than these raw materials.So the present invention desires to provide a kind of method of from Hemp Eupatorium, effectively extracting beta-elemene, for the production cost and the exploitation that reduce beta-elemene significantly, utilize, administer this worldwide malignant weed plant of Hemp Eupatorium and opened up new approach.
Summary of the invention
The technical problem that the present invention solved provide a kind of from Hemp Eupatorium the method for extraction separation beta-elemene.
Detect discovery through the contriver: the beta-elemene in the Hemp Eupatorium mainly is distributed in its leaf and the stem and (accounts for more than 90% of herb total amount).Wherein, with content in the leaf the highest (being about 0.4-0.6%), and content is extremely low or do not have basically in other organs such as root, flower.Comparative analysis is the result show, beta-elemene content is far above the content of Chinese medicine curcuma zedoary, myrrh, lemongrass, Herba Solidaginis in the Hemp Eupatorium leaf.
Employed raw material is mainly the stem of Hemp Eupatorium, at least a or its mixing in the leaf in the process for extracting of the present invention; Can not influence the extraction separation beta-elemene if be mixed with other organs such as root, flower, fruit in the raw material yet; But can influence the yield of process for extracting, so preferably be mixed into raw material with at least a in the stem of Hemp Eupatorium, the leaf or its.
Process for extracting of the present invention is following:
A, raw material prepare: mainly be mixed into raw material with at least a in the stem of Hemp Eupatorium, the leaf or its, bright article are pulled an oar or drying and crushing;
B, volatile oil extract: from steps A gained raw material, extract and obtain volatile oil;
C, column chromatography: the volatile oil that step B is obtained carries out column chromatography, collects elutriant, removes the solvent in the elutriant, obtains the beta-elemene bullion.
Further, the beta-elemene bullion that volatile oil that can also step B be obtained or step C obtain carries out further refining, obtains the higher beta-elemene highly finished product of purity.
Be that purification step D is: the beta-elemene bullion that volatile oil that step B is obtained or step C obtain, adopt short-path distillation or preparative high performance liquid chromatography system make with extra care the highly purified beta-elemene of acquisition.
Adopting aforesaid method to handle the stem of Hemp Eupatorium, leaf step B extracts and can obtain beta-elemene content usually greater than 20% volatile oil; Adopt method such as column chromatography to handle and can obtain beta-elemene content usually greater than 40%, even up to 60% beta-elemene bullion; Finally, further refined volatile oil or beta-elemene bullion are collected the purifying composition that contains beta-elemene according to " target content ", can obtain beta-elemene content usually greater than 90%, even up to 98% beta-elemene highly finished product.
Can adopt one of following five kinds of methods to extract volatile oil when wherein, extracting volatile oil:
Method one: steam distillation:
(1) raw material through processing of step A carries out air distillation, obtains distillate;
(2) in distillate, add boiling point less than 60 ℃ non-polar organic solvent, upper organic phase is collected in extraction, removes the organic solvent in the upper organic phase, promptly gets volatile oil.Wherein, preferably primary solvent during distillation with water, add and improve the extraction system boiling point and prevent the blistered material of saponification.As, adding terepthaloyl moietie is the temperature that improves distillation azeotropic liquid, adding ethanol is to eliminate the saponification foaming phenomenon that produces in the distillation, adds zeolite and be bumping when preventing to distill.So existing, known, conventional, foreseeable, material replacement terepthaloyl moietie with above-mentioned function capable of using, ethanol, zeolite are to reach identical purpose.
Method two: pickling process:
(1) raw material through processing of step A adds at least a or its arbitrary proportion blended organic solvent lixiviate, filtration in acetone, sherwood oil or the methyl alcohol, gets vat liquor;
(2) vat liquor removes the ethanol that the back of desolvating adds concentration 20-80%v/v, and the adding boiling point extracts less than 60 ℃ non-polar organic solvent, collects upper organic phase, removes the organic solvent in the upper organic phase, promptly gets volatile oil.
Method three: Continuous Countercurrent Extraction method:
(1) raw material through processing of step A adds at least a or its arbitrary proportion blended organic solvent Continuous Countercurrent Extraction in acetone, sherwood oil or the methyl alcohol, gets extracting solution;
(2) ethanol of adding concentration 20-80%v/v behind the solvent in the removal extracting solution, the adding boiling point extracts less than 60 ℃ non-polar organic solvent, collects upper organic phase, removes the organic solvent in the upper organic phase, promptly gets volatile oil.
Method four: microwave extraction method:
(1) raw material through processing of step A adds at least a or its arbitrary proportion blended organic solvent in acetone, sherwood oil or the methyl alcohol, in microwave extracting apparatus, adopts power 300-800W extraction 300-800s, gets extracting solution;
(2) ethanol of adding concentration 20-80%v/v behind the extracting solution removal solvent, the adding boiling point extracts less than 60 ℃ non-polar organic solvent, collects upper organic phase, removes the organic solvent in the upper organic phase, promptly gets volatile oil.
Method five: supercritical extraction:
(1) through the raw material of processing of step A, uses supercritical CO
2The fluid extraction device extracts, extracting pressure 20-30MPa, and extraction temperature 30-50 ℃, extraction time 1.5-3h gets extraction product;
(2) ethanol of adding concentration 20-80%v/v in the extraction product, the adding boiling point extracts less than 60 ℃ non-polar organic solvent, collects upper organic phase, removes the organic solvent in the upper organic phase, promptly gets volatile oil.
Adopting boiling point during above-mentioned five kinds of volatile oil extraction methods extraction is because if boiling point is higher than 60 ℃ less than 60 ℃ non-polar organic solvent; Remove and can beta-elemene be removed in the lump when desolvating, so the selection boiling point is less than 60 ℃ non-polar organic solvent extracting and separating beta-elemene.
Wherein, the adoptable method of step C column chromatography is following:
Method 1: macroporous resin chromatography column on the volatile oil that step B is obtained, at least a or its arbitrary proportion blended solvent elution in water, methyl alcohol, ethanol, the acetone, flow velocity is 1-4% column volume/min; Collection contains the elutriant of beta-elemene, removes solvent, behind the ethanol of adding concentration 20-80%v/v; With the non-polar organic solvent extraction of boiling point less than 60 ℃; Collect upper organic phase, remove the organic solvent in the upper organic phase, promptly get the beta-elemene bullion.
Or employing method 2: silica gel column chromatography or silver ions silica gel column chromatography on the volatile oil that step B is obtained; With the non-polar organic solvent wash-out of boiling point less than 60 ℃, flow velocity is 1-4% column volume/min, collects the elutriant that contains beta-elemene; Remove and desolvate, obtain the beta-elemene bullion.
Wherein, when obtaining the beta-elemene highly finished product, the bullion that volatile oil that can step B be obtained or step D obtain adopts one of following method to make with extra care, and obtains highly purified beta-elemene.Normally collect the purifying composition contain beta-elemene when refining, can obtain beta-elemene content usually greater than 90% according to " target content ", even up to 98% beta-elemene highly finished product.
Process for purification 1: during molecular distillation method: the beta-elemene bullion that volatile oil that step B is obtained or step C obtain carries out short-path distillation, collects the distillate that beta-elemene reaches target content, promptly gets the beta-elemene highly finished product.
Process for purification 2: highly effective liquid phase chromatographic system preparation method: the beta-elemene bullion that volatile oil that step B is obtained or step C obtain separates through the preparative high performance liquid chromatography system; Collect the effluent that beta-elemene reaches target content, removing desolvates promptly gets the beta-elemene highly finished product.
Process for extracting gained beta-elemene elaboration purity of the present invention is at least greater than 90%; Even can be up to 98%; Productive rate can be up to 70%; For effectively utilizing malignant weed plant Hemp Eupatorium that a kind of new approach is provided, more obtain beta-elemene a kind of brand-new, cheap, effective means is provided.
Embodiment
The contriver adopts the performance liquid chromatography micrometric measurement to find: the beta-elemene in the Hemp Eupatorium mainly is distributed in its leaf and the stem and (accounts for more than 90% of herb total amount).Wherein, with content in the leaf the highest (being about 0.4-0.6%), and content is extremely low or do not have basically in other organs such as root, flower.And comparative analysis is the result show, beta-elemene content is far above the content of Chinese medicine curcuma zedoary, myrrh, lemongrass, Herba Solidaginis in the Hemp Eupatorium leaf.
Process for extracting of the present invention is following:
A, raw material prepare: with stem, the leaf of Hemp Eupatorium at least a or its be mixed into raw material, bright article are pulled an oar or drying and crushing;
B, extraction volatile oil: from steps A gained raw material, obtain volatile oil;
C, column chromatography: the volatile oil that step B is obtained carries out column chromatography, collects elutriant, removes solvent in the elutriant, obtains the beta-elemene bullion.
Purification step D is: the beta-elemene bullion that volatile oil that step B is obtained or step C obtain, adopt short-path distillation or preparative high performance liquid chromatography system make with extra care the highly purified beta-elemene of acquisition.
Can adopt one of following five kinds of methods to extract volatile oil when one, extracting volatile oil:
Method one, steam distillation: adding entry through the raw material of processing of step A is that main solvent carries out air distillation, obtains containing the distillate of beta-elemene; In the gained distillate, add boiling points such as sherwood oil, hexanaphthene, normal hexane, benzene, ether less than 60 ℃ non-polar organic solvent, extract at least 2 times, collect upper organic phase, remove the organic solvent in the upper organic phase, promptly get volatile oil.Wherein, preferably primary solvent during distillation with water, add terepthaloyl moietie, ethanol.Because of regional reason boiling point is low, add the temperature that terepthaloyl moietie can improve distillation azeotropic liquid; Adding ethanol is the saponification foaming phenomenon that produces in the distillation in order to eliminate; Also can add zeolite and be bumping when preventing to distill.So existing, known, conventional, foreseeable, material replacement terepthaloyl moietie with above-mentioned function capable of using, ethanol, zeolite are to reach identical purpose.
Method two, pickling process: the raw material through processing of step A adds organic solvent lixiviates 2-4 time such as acetone, sherwood oil or methyl alcohol, filters; United extraction liquid is also removed solvent; The ethanolic soln that adds 20-80% with organic solvent extractions such as sherwood oil at least 2 times, is collected upper organic phase; Remove the organic solvent in the upper organic phase, promptly get volatile oil.
Method three, Continuous Countercurrent Extraction method: the raw material through processing of step A adds organic solvent Continuous Countercurrent Extraction such as acetone, sherwood oil or methyl alcohol 3-4 time, obtains extracting solution.Remove the solvent in the extracting solution; The ethanolic soln that adds 20-80% extracts 2 times with the non-polar organic solvent of boiling points such as sherwood oil, hexanaphthene, normal hexane, benzene, ether less than 60 ℃ at least, collects upper organic phase; Remove the organic solvent in the upper organic phase, promptly get volatile oil.
Method four, microwave extraction method: the raw material through processing of step A adds organic solvents such as acetone, sherwood oil or methyl alcohol, and in microwave extracting apparatus, the power of 300-800W is extraction 300-800s down, extracts 2-4 time, filters united extraction liquid.Remove the solvent in the extracting solution; The ethanolic soln that adds 20-80% extracts 2 times with the non-polar organic solvent of boiling points such as sherwood oil, hexanaphthene, normal hexane, benzene, ether less than 60 ℃ at least, collects upper organic phase; Remove the organic solvent in the upper organic phase, promptly get volatile oil.
Method five, supercritical extraction:, use supercritical CO through the raw material of processing of step A
2The fluid extraction device extracts, extracting pressure 20-30MPa, extraction temperature 30-50 ℃, extraction time 1.5-3h.In extraction product, add the ethanolic soln of 20-80%, with the non-polar organic solvent of boiling points such as sherwood oil, hexanaphthene, normal hexane, benzene, ether less than 60 ℃; At least extract 2 times; Collect upper organic phase, remove the organic solvent in the upper organic phase, promptly get volatile oil.
Through comparing, above-mentioned five kinds of methods all can obtain volatile oil (beta-elemene content＞20%) from raw material.But best process for extracting is Continuous Countercurrent Extraction method and steam distillation, wherein: and advantages such as the Continuous Countercurrent Extraction method has the extraction yield height, and solvent load is less, but beta-elemene content is lower in the extraction gained volatile oil; That steam distillation has is easy and simple to handle, extract beta-elemene content advantage of higher in the gained volatile oil, and volatile oil purity is generally 30%, even can reach 40%, but extraction yield is on the low side.
Wherein, the Continuous Countercurrent Extraction condition is: the acetone that adds material quantity (g) 3-7 times of volume (mL) is made solvent, soaks 1h at least under the room temperature, and constant temperature (20-30 ℃) gas bath vibration (100-140r/min) Continuous Countercurrent Extraction at least 1 time is no less than 1.5h at every turn.Parameters optimization is: the acetone that adds material quantity (g) 5 times of volumes (mL) is made solvent, soaks 4h under the room temperature, constant temperature (25 ℃) gas bath vibration (120r/min) Continuous Countercurrent Extraction 3 times, the 1st extraction 2.5h, the 2nd extraction 3.5h, the 3rd extraction 3h.
The steam distillation extraction conditions is: get the raw material that steps A obtains, put into matrass, add terepthaloyl moietie, small amount of ethanol, zeolite that water, 1-3 that material quantity (g) 3-7 doubly measures (mL) doubly measure (mL), 80-150 ℃ distillate is collected in the air distillation of heating.Parameters optimization is: get the raw material that steps A obtains, put into matrass, add the water of material quantity (g) 5 times of amounts (mL), terepthaloyl moietie, small amount of ethanol, the zeolite of 2 times of amounts (mL), 80-150 ℃ distillate is collected in the air distillation of heating.
Two, column chromatography method is following:
During column chromatography, the inventor takes silica gel, silver ions silica gel, D101 macroporous resin, HP20 macroporous resin to do chromatography media all can from the volatile oil of Hemp Eupatorium, to separate and obtain the beta-elemene bullion, beta-elemene purity usually can＞40%.Adoptable method is following:
Method one, Amberlyst process: macroporous resin chromatography column on the volatile oil that step B is obtained, water, methyl alcohol, ethanol, acetone equal solvent or its mixing solutions wash-out, flow velocity is 1-4% column volume/min; Collection contains the elutriant of beta-elemene, removes solvent, adds the ethanolic soln of 20-80%; With organic solvent extractions such as sherwood oil at least 2 times; Separate upper organic phase, remove the organic solvent in the upper organic phase, promptly get the beta-elemene bullion.
Method two, silica gel method: silica gel column chromatography or silver ions silica gel column chromatography on the volatile oil that step B is obtained, use the sherwood oil wash-out, flow velocity is 1-4% column volume/min, collects the elutriant that contains beta-elemene, removes and desolvates, and obtains the beta-elemene bullion.
Concrete, the chromatography condition of optimization is: be chromatography media with HP20, successively carry out wash-out, flow velocity 2% column volume/min with zero(ppm) water, 80% ethanol, 85% ethanol, 90% ethanol, 95% ethanol, the absolute ethyl alcohol of 2 times of column volumes respectively.Carry out column chromatography by this condition, the purity of beta-elemene bullion＞60%, the recovery＞85%.
Three, process for purification is following:
When refining, the contriver finds to take molecular distillation method, highly effective liquid phase chromatographic system preparation method all can from volatile oil or bullion, separate obtaining highly purified beta-elemene.
The short-path distillation preferable methods is: the bullion that volatile oil that step B is obtained or step C obtain carries out vacuum fractionation, collects the cut of 76-78 ℃/40Pa, is pure article (purity＞90%, productive rate＞70%).
Highly effective liquid phase chromatographic system prepares one of preferable methods: the bullion that volatile oil that step B is obtained or step C obtain separates with following highly effective liquid phase chromatographic system: Tianjin, island high performance liquid chromatograph LC-10A; The C18 preparative column (10mm * 150mm); Eluent is ethanol/water/acetonitrile=70/20/10 (V/V); Flow velocity is 2mL/min, and the detection wavelength is 210nm; Sampling volume 1mL.Collect the beta-elemene effluent, add sherwood oil, extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, add Na
2SO
4Dry 10-24h, solvent is drained in 60 ℃ of vacuum rotations, obtains pure article (purity＞98%, productive rate＞75%).
Below prepare instance through the present invention beneficial effect of the present invention be described.
Embodiment 1 distillation method extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect Hemp Eupatorium stem and leaf, dry naturally under the sun, pulverize, cross 40 mesh sieves.
The extraction of B, volatile oil: get the raw material 100g that steps A obtains, put into matrass, add deionized water or zero(ppm) water 500mL, terepthaloyl moietie 200mL, ethanol 50mL, zeolite 3-5 grain, 80-150 ℃ distillate is collected in the air distillation of heating.In the distillate that obtains, add sherwood oil (boiling range 30-60 ℃) in 1: 1 ratio, extract repeatedly three times.Separate and merge petroleum ether layer, solvent is drained in 60 ℃ of vacuum rotations, obtains volatile oil (purity＞40%).
C, performance liquid preparation: the volatile oil that step B is obtained separates with following highly effective liquid phase chromatographic system: Tianjin, island high performance liquid chromatograph LC-10A; The C18 preparative column (10mm * 150mm); Eluent is ethanol/water/acetonitrile=70/20/10 (V/V); Flow velocity is 2mL/min, and the detection wavelength is 210nm; Sampling volume 1mL.Collect the beta-elemene effluent, add sherwood oil, extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, add Na
2SO
4Dry 10-24h, solvent is drained in 60 ℃ of vacuum rotations, obtains pure article (purity＞98%, productive rate＞75%).
Embodiment 2 distillation method extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect Hemp Eupatorium stem and leaf, break into pulpous state with kibbler.
The extraction of B, volatile oil: get the raw material 100g that steps A obtains, put into matrass, add deionized water or zero(ppm) water 500mL, terepthaloyl moietie 200mL, ethanol 50mL, zeolite 3-5 grain, 80-150 ℃ distillate is collected in the air distillation of heating.In the distillate that obtains, add sherwood oil (boiling range 30-60 ℃) in 1: 1.5 ratio, extract repeatedly three times.Separate and merge petroleum ether layer, solvent is drained in 60 ℃ of vacuum rotations, obtains volatile oil (purity＞35%).
C, performance liquid preparation: the volatile oil that step B is obtained separates with following highly effective liquid phase chromatographic system: Tianjin, island high performance liquid chromatograph LC-10A; The C18 preparative column (10mm * 150mm); Eluent is ethanol/water/acetonitrile=70/20/10 (V/V); Flow velocity is 2mL/min, and the detection wavelength is 210nm; Sampling volume 1mL.Collect the beta-elemene effluent, add sherwood oil, extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, add Na
2SO
4Dry 10-24h, solvent is drained in 60 ℃ of vacuum rotations, obtains pure article (purity＞97%, productive rate＞73%).
Embodiment 3 distillation method extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect Hemp Eupatorium stem and leaf, break into pulpous state with kibbler.
The extraction of B, volatile oil: get the raw material 100g that steps A obtains, put into matrass, add deionized water or zero(ppm) water 500mL, terepthaloyl moietie 200mL, ethanol 50mL, zeolite 3-5 grain, 80-150 ℃ distillate is collected in the air distillation of heating.In the distillate that obtains, add hexanaphthene in 1: 1 ratio, extract repeatedly three times.Separate and merge petroleum ether layer, solvent is drained in 60 ℃ of vacuum rotations, obtains volatile oil (purity＞37%).
C, column chromatography: with the volatile oil that step B obtains, last 300mL HP20 macroporous resin chromatography column successively carries out wash-out, flow velocity 6mL/min with zero(ppm) water, 80% ethanol, 85% ethanol, 90% ethanol, 95% ethanol, the absolute ethyl alcohol of 600mL respectively.Collect the elutriant of beta-elemene content＞60%, add sherwood oil (boiling range 30-60 ℃), extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, solvent is drained in 60 ℃ of vacuum rotations, obtains bullion (purity＞60%).
D, short-path distillation: the bullion that step C is obtained carries out vacuum fractionation, collects the cut of 76-78 ℃/40Pa, is pure article (purity＞90%, productive rate＞73%).
Embodiment 4 distillation method extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect Hemp Eupatorium stem and leaf, break into pulpous state with kibbler.
The extraction of B, volatile oil: get the raw material 100g that steps A obtains, put into matrass, add deionized water or zero(ppm) water 500mL, terepthaloyl moietie 200mL, ethanol 50mL, zeolite 3-5 grain, 80-150 ℃ distillate is collected in the air distillation of heating.In the distillate that obtains, add sherwood oil (boiling range 30-60 ℃) in 1: 1 ratio, extract repeatedly three times.Separate and merge petroleum ether layer, solvent is drained in 60 ℃ of vacuum rotations, obtains volatile oil (purity＞35%).
C, short-path distillation: the volatile oil that step B is obtained carries out vacuum fractionation, collects the cut of 76-78 ℃/40Pa, is pure article (purity＞90%, productive rate＞70%).
Embodiment 5 Continuous Countercurrent Extraction methods extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect fresh Hemp Eupatorium stem and leaf, break into pulpous state with kibbler.
B, Continuous Countercurrent Extraction: get steps A raw material 100g, put into triangular flask, Continuous Countercurrent Extraction three times adds acetone 500mL at every turn, and 4-8h, temperature 20-25 ℃ are extracted in concussion (rotating speed 60-100r/min).Filter united extraction liquid, concentrating under reduced pressure removes and desolvates, and adds 50% ethanol 100mL, uses 100mL sherwood oil (boiling range 30-60 ℃) extraction 3 times again.Separate and merge petroleum ether layer, 60 ℃ of vacuum rotation solvent evaporated obtain volatile oil (purity＞20%).
C, column chromatography: with the volatile oil that step B obtains, last 300mL HP20 macroporous resin chromatography column successively carries out wash-out, flow velocity 6mL/min with zero(ppm) water, 80% ethanol, 85% ethanol, 90% ethanol, 95% ethanol, the absolute ethyl alcohol of 600mL respectively.Collect the elutriant of beta-elemene content＞60%, add sherwood oil (boiling range 30-60 ℃), extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, solvent is drained in 60 ℃ of vacuum rotations, obtains bullion (purity＞60%).
D, short-path distillation: the bullion that step C is obtained carries out vacuum fractionation, collects the cut of 76-78 ℃/40Pa, is pure article (purity＞90%, productive rate＞70%).
Embodiment 6 Continuous Countercurrent Extraction methods extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect fresh Hemp Eupatorium stem and leaf, dry naturally under the sun, pulverize, cross 40 mesh sieves.
B, Continuous Countercurrent Extraction: get steps A raw material 100g, put into triangular flask, Continuous Countercurrent Extraction three times adds sherwood oil 500mL at every turn, and 4-8h, temperature 20-25 ℃ are extracted in concussion (rotating speed 60-100r/min).Filter united extraction liquid, concentrating under reduced pressure removes and desolvates, and adds 50% ethanol 100mL, uses 100mL sherwood oil (boiling range 30-60 ℃) extraction 3 times again.Separate and merge petroleum ether layer, 60 ℃ of vacuum rotation solvent evaporated obtain volatile oil (purity＞20%).
C, column chromatography: with the volatile oil that step B obtains, last 300mL HP20 macroporous resin chromatography column successively carries out wash-out, flow velocity 6mL/min with zero(ppm) water, 80% ethanol, 85% ethanol, 90% ethanol, 95% ethanol, the absolute ethyl alcohol of 600mL respectively.Collect the elutriant of beta-elemene content＞60%, add sherwood oil (boiling range 30-60 ℃), extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, solvent is drained in 60 ℃ of vacuum rotations, obtains bullion (purity＞60%).
D, short-path distillation: the bullion that step C is obtained carries out vacuum fractionation, collects the cut of 76-78 ℃/40Pa, is pure article (purity＞90%, productive rate＞70%).
Embodiment 7 Continuous Countercurrent Extraction methods extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect fresh Hemp Eupatorium stem and leaf, dry naturally under the sun, pulverize, cross 40 mesh sieves.
B, Continuous Countercurrent Extraction: get steps A raw material 100g, put into triangular flask, Continuous Countercurrent Extraction three times adds methyl alcohol 500mL at every turn, and 4-8h, temperature 20-25 ℃ are extracted in concussion (rotating speed 60-100r/min).Filter united extraction liquid, concentrating under reduced pressure removes and desolvates, and adds 50% ethanol 100mL, uses 100mL sherwood oil (boiling range 30-60 ℃) extraction 3 times again.Separate and merge petroleum ether layer, 60 ℃ of vacuum rotation solvent evaporated obtain volatile oil (purity＞20%).
C, column chromatography: with the volatile oil that step B obtains, last 300mL HP20 macroporous resin chromatography column successively carries out wash-out, flow velocity 6mL/min with zero(ppm) water, 80% ethanol, 85% ethanol, 90% ethanol, 95% ethanol, the absolute ethyl alcohol of 600mL respectively.Collect the elutriant of beta-elemene content＞60%, add sherwood oil (boiling range 30-60 ℃), extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, solvent is drained in 60 ℃ of vacuum rotations, obtains bullion (purity＞60%).
D, short-path distillation: the bullion that step C is obtained carries out vacuum fractionation, collects the cut of 76-78 ℃/40Pa, is pure article (purity＞90%, productive rate＞70%).
Embodiment 8 Continuous Countercurrent Extraction methods extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect fresh Hemp Eupatorium stem and leaf, dry naturally under the sun, pulverize, cross 40 mesh sieves.
B, Continuous Countercurrent Extraction: get steps A raw material 100g, put into triangular flask, Continuous Countercurrent Extraction three times adds methyl alcohol 500mL at every turn, and 4-8h, temperature 20-25 ℃ are extracted in concussion (rotating speed 60-100r/min).Filter united extraction liquid, concentrating under reduced pressure removes and desolvates, and adds 50% ethanol 100mL, uses 100mL sherwood oil (boiling range 30-60 ℃) extraction 3 times again.Separate and merge petroleum ether layer, 60 ℃ of vacuum rotation solvent evaporated obtain volatile oil (purity＞20%).
C, performance liquid preparation: the volatile oil that step B is obtained separates with following highly effective liquid phase chromatographic system: Tianjin, island high performance liquid chromatograph LC-10A; The C18 preparative column (10mm * 150mm); Eluent is ethanol/water/acetonitrile=70/20/10 (V/V); Flow velocity is 2mL/min, and the detection wavelength is 210nm; Sampling volume 1mL.Collect the beta-elemene effluent, add sherwood oil, extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, add Na
2SO
4Dry 10-24h, solvent is drained in 60 ℃ of vacuum rotations, obtains pure article (purity＞98%, productive rate＞75%).
Embodiment 9 pickling processes extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect fresh Hemp Eupatorium stem and leaf, dry naturally under the sun, pulverize, cross 40 mesh sieves.
B, pickling process: the raw material 100g of processing of step A,, add the 500ml acetone extraction 3 times, each 2 hours, filter, united extraction liquid is also removed acetone, adds 50% ethanol 100mL, uses 100mL sherwood oil (boiling range 30-60 ℃) extraction 2 times again.Separate and merge petroleum ether layer, 60 ℃ of vacuum rotation solvent evaporated obtain volatile oil (purity＞20%).
C, performance liquid preparation: the volatile oil that step B is obtained separates with following highly effective liquid phase chromatographic system: Tianjin, island high performance liquid chromatograph LC-10A; The C18 preparative column (10mm * 150mm); Eluent is ethanol/water/acetonitrile=70/20/10 (V/V); Flow velocity is 2mL/min, and the detection wavelength is 210nm; Sampling volume 1mL.Collect the beta-elemene effluent, add sherwood oil, extract repeatedly three times in 1: 1 ratio.Separate and merge petroleum ether layer, add Na
2SO
4Dry 10-24h, solvent is drained in 60 ℃ of vacuum rotations, obtains pure article (purity＞98%, productive rate＞75%).
Wherein, when step B adopted sherwood oil or methyl alcohol lixiviate, pure article purity and productive rate were suitable with the employing acetone extraction.
Embodiment 10 microwave extraction methods extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect fresh Hemp Eupatorium stem and leaf, dry naturally under the sun, pulverize, cross 40 mesh sieves.
B, microwave extracting: the raw material 100g of processing of step A,, in microwave extracting apparatus, power 800W extracts 400s, adds acetone extract 3 times, filters united extraction liquid.Remove the solvent in the extracting solution, add 50% ethanol 100mL, use 100mL sherwood oil (boiling range 30-60 ℃) extraction 2 times again.Separate and merge petroleum ether layer, 60 ℃ of vacuum rotation solvent evaporated obtain volatile oil (purity＞20%).
C, short-path distillation: the volatile oil that step B is obtained carries out vacuum fractionation, collects the cut of 76-78 ℃/40Pa, is pure article (purity＞90%, productive rate＞70%).
Wherein, when step B adopted sherwood oil or methyl alcohol lixiviate, pure article purity and productive rate were suitable with the employing acetone extraction.
Embodiment 11 supercritical extractions extraction separation beta-elemene from Hemp Eupatorium
A, raw material are prepared: collect fresh Hemp Eupatorium stem and leaf, dry naturally under the sun, pulverize, cross 40 mesh sieves.
B, supercritical extraction: the raw material 100g of processing of step A,, use supercritical CO
2The fluid extraction device extracts, control extracting pressure 20-30MPa, extraction temperature 30-50 ℃, extraction time 2h.In extraction product, add 50% ethanol 100mL, use 100mL sherwood oil (boiling range 30-60 ℃) extraction 2 times again.Separate and merge petroleum ether layer, 60 ℃ of vacuum rotation solvent evaporated obtain volatile oil (purity＞20%).
C, short-path distillation: the volatile oil that step B is obtained carries out vacuum fractionation, collects the cut of 76-78 ℃/40Pa, is pure article (purity＞90%, productive rate＞70%).
Claims (10)
1. the method for extraction separation beta-elemene from Hemp Eupatorium, it is characterized in that: step is following:
A, raw material prepare: mainly be mixed into raw material with at least a in the stem of Hemp Eupatorium, the leaf or its, bright article are pulled an oar or drying and crushing;
B, volatile oil extract: from steps A gained raw material, extract and obtain volatile oil;
C, column chromatography: the volatile oil that step B is obtained carries out column chromatography, collects elutriant, removes the solvent in the elutriant, obtains the beta-elemene bullion.
2. the method for extraction separation beta-elemene from Hemp Eupatorium, it is characterized in that: step is following:
A, raw material prepare: mainly be mixed into raw material with at least a in the stem of Hemp Eupatorium, the leaf or its, bright article are pulled an oar or drying and crushing;
B, volatile oil extract: from steps A gained raw material, extract and obtain volatile oil;
C, column chromatography: the volatile oil that step B is obtained carries out column chromatography, collects elutriant, removes the solvent in the elutriant, obtains the beta-elemene bullion;
D, refining: the beta-elemene bullion that volatile oil that step B is obtained or step C obtain, adopt short-path distillation or preparative high performance liquid chromatography system make with extra care acquisition beta-elemene highly finished product.
3. according to claim 1 and 2 from Hemp Eupatorium the method for extraction separation beta-elemene, it is characterized in that: when step B adopted wet distillation to extract volatile oil, method was following:
(1) raw material through processing of step A carries out air distillation, obtains distillate;
(2) in distillate, add boiling point less than 60 ℃ non-polar organic solvent, upper organic phase is collected in extraction, removes the organic solvent in the upper organic phase, promptly gets volatile oil;
Be primary solvent with water preferably during distillation, add in the solvent systems and improve the solvent systems boiling point and prevent saponification foaming, the material of bumping.
4. according to claim 1 and 2 from Hemp Eupatorium the method for extraction separation beta-elemene, it is characterized in that: when step B adopted pickling process to extract volatile oil, method was following:
(1) raw material through processing of step A adds at least a or its arbitrary proportion blended organic solvent lixiviate, filtration in acetone, sherwood oil or the methyl alcohol, gets vat liquor;
(2) vat liquor removes the ethanol that the back of desolvating adds concentration 20-80%v/v, and the adding boiling point extracts less than 60 ℃ non-polar organic solvent, collects upper organic phase, removes the organic solvent in the upper organic phase, promptly gets volatile oil.
5. according to claim 1 and 2 from Hemp Eupatorium the method for extraction separation beta-elemene, it is characterized in that: when step B adopted the Continuous Countercurrent Extraction method to extract volatile oil, method was following:
(1) raw material through processing of step A adds at least a or its arbitrary proportion blended organic solvent Continuous Countercurrent Extraction in acetone, sherwood oil or the methyl alcohol, gets extracting solution;
(2) ethanol of adding concentration 20-80%v/v behind the solvent in the removal extracting solution, the adding boiling point extracts less than 60 ℃ non-polar organic solvent, collects upper organic phase, removes the organic solvent in the upper organic phase, promptly gets volatile oil.
6. according to claim 1 and 2 from Hemp Eupatorium the method for extraction separation beta-elemene, it is characterized in that: when step B adopted microwave extraction method to extract volatile oil, method was following:
(1) raw material through processing of step A adds at least a or its arbitrary proportion blended organic solvent in acetone, sherwood oil or the methyl alcohol, in microwave extracting apparatus, adopts power 300-800W extraction 300-800s, gets extracting solution;
(2) ethanol of adding concentration 20-80%v/v behind the extracting solution removal solvent, the adding boiling point extracts less than 60 ℃ non-polar organic solvent, collects upper organic phase, removes the organic solvent in the upper organic phase, promptly gets volatile oil.
7. according to claim 1 and 2 from Hemp Eupatorium the method for extraction separation beta-elemene, it is characterized in that: when step B adopted supercritical fluid extraction volatile oil, method was following:
(1) through the raw material of processing of step A, extract with the supercritical carbon dioxide extraction device, extracting pressure 20-30MPa, extraction temperature 30-50 ℃, extraction time 1.5-3h gets extraction product;
(2) ethanol of adding concentration 20-80%v/v in the extraction product, the adding boiling point extracts less than 60 ℃ non-polar organic solvent, collects upper organic phase, removes the organic solvent in the upper organic phase, promptly gets volatile oil.
8. according to claim 1 and 2 from Hemp Eupatorium the method for extraction separation beta-elemene, it is characterized in that: column chromatography adopts following method separation and purification among the step C:
Method 1: macroporous resin chromatography column on the volatile oil that step B is obtained, at least a or its arbitrary proportion blended solvent elution in water, methyl alcohol, ethanol, the acetone, flow velocity is 1-4% column volume/min; Collection contains the elutriant of beta-elemene, removes solvent, behind the ethanol of adding concentration 20-80%v/v; With the non-polar organic solvent extraction of boiling point less than 60 ℃; Collect upper organic phase, remove the organic solvent in the upper organic phase, promptly get the beta-elemene bullion;
Or method 2: silica gel column chromatography or silver ions silica gel column chromatography on the volatile oil that step B is obtained; With the non-polar organic solvent wash-out of boiling point less than 60 ℃, flow velocity is 1-4% column volume/min, collects the elutriant that contains beta-elemene; Remove and desolvate, obtain the beta-elemene bullion.
9. according to claim 2 from Hemp Eupatorium the method for extraction separation beta-elemene; It is characterized in that: when step D adopts molecular distillation method; Method is following: the beta-elemene bullion that volatile oil that step B is obtained or step C obtain carries out short-path distillation; Collect the distillate that beta-elemene reaches target content, promptly get the beta-elemene highly finished product.
10. beta-elemene process for purification according to claim 2; It is characterized in that: when step D adopts the highly effective liquid phase chromatographic system preparation method; Method is following: the beta-elemene bullion that volatile oil that step B is obtained or step C obtain separates through the preparative high performance liquid chromatography system; Collect the effluent that beta-elemene reaches target content, the solvent of removing in the effluent promptly gets the beta-elemene highly finished product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110457523.5A CN102432419B (en) | 2011-12-31 | 2011-12-31 | Method for extracting and separating beta-elemene from Eupatorium adenophorum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201110457523.5A CN102432419B (en) | 2011-12-31 | 2011-12-31 | Method for extracting and separating beta-elemene from Eupatorium adenophorum |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102432419A true CN102432419A (en) | 2012-05-02 |
| CN102432419B CN102432419B (en) | 2014-05-14 |
Family
ID=45980815
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201110457523.5A Active CN102432419B (en) | 2011-12-31 | 2011-12-31 | Method for extracting and separating beta-elemene from Eupatorium adenophorum |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102432419B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106480004A (en) * | 2016-10-27 | 2017-03-08 | 杭州师范大学 | A kind of sesquiterpene synthase in Eupatorium adenophorum source, gene, carrier, engineering bacteria and its application |
| CN112535180A (en) * | 2020-11-30 | 2021-03-23 | 湖南人文科技学院 | Eupatorium adenophorum spreng extract and application thereof in preventing and controlling paddy field weeds |
| CN114668072A (en) * | 2022-03-10 | 2022-06-28 | 昆明悦馨生物科技有限公司 | Comprehensive utilization method of Eupatorium adenophorum |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6464839B1 (en) * | 1997-04-14 | 2002-10-15 | Yuan Da International Group Limited | Beta-elemene, method to prepare the same and uses thereof |
| CN101402544A (en) * | 2008-11-14 | 2009-04-08 | 沈阳万爱普利德医药科技有限公司 | Industrial production method of high-purity beta-elemi alkene bulk medicament |
| CN101402543A (en) * | 2008-11-14 | 2009-04-08 | 沈阳万爱普利德医药科技有限公司 | Beta-elemi alkene bulk medicament and method of preparing its preparations |
-
2011
- 2011-12-31 CN CN201110457523.5A patent/CN102432419B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6464839B1 (en) * | 1997-04-14 | 2002-10-15 | Yuan Da International Group Limited | Beta-elemene, method to prepare the same and uses thereof |
| CN101402544A (en) * | 2008-11-14 | 2009-04-08 | 沈阳万爱普利德医药科技有限公司 | Industrial production method of high-purity beta-elemi alkene bulk medicament |
| CN101402543A (en) * | 2008-11-14 | 2009-04-08 | 沈阳万爱普利德医药科技有限公司 | Beta-elemi alkene bulk medicament and method of preparing its preparations |
Non-Patent Citations (2)
| Title |
|---|
| 《安徽农业科学》 20111110 韦会平等 "紫茎泽兰不同生长部位中beta-榄香烯的含量测定" 第19768-19769,19870页 1-10 第39卷, 第32期 * |
| 韦会平等: ""紫茎泽兰不同生长部位中β-榄香烯的含量测定"", 《安徽农业科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106480004A (en) * | 2016-10-27 | 2017-03-08 | 杭州师范大学 | A kind of sesquiterpene synthase in Eupatorium adenophorum source, gene, carrier, engineering bacteria and its application |
| CN106480004B (en) * | 2016-10-27 | 2019-05-24 | 杭州师范大学 | A kind of sesquiterpene synthase, gene, carrier, engineering bacteria and its application in Eupatorium adenophorum source |
| CN112535180A (en) * | 2020-11-30 | 2021-03-23 | 湖南人文科技学院 | Eupatorium adenophorum spreng extract and application thereof in preventing and controlling paddy field weeds |
| CN114668072A (en) * | 2022-03-10 | 2022-06-28 | 昆明悦馨生物科技有限公司 | Comprehensive utilization method of Eupatorium adenophorum |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102432419B (en) | 2014-05-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109942380B (en) | Method for preparing cannabidiol by high-speed counter-current chromatography separation and purification | |
| CN101336949B (en) | Method for extracting polysaccharide and flavone from Gynura divaricata | |
| CN111960930A (en) | Method for separating and purifying cannabidiol from industrial cannabis sativa leaves | |
| CN102976909A (en) | Method for extracting and purifying 6-gingerol from ginger | |
| CN104892687B (en) | The method that high speed adverse current chromatogram isolates and purifies monomeric compound in Chinese mahonia leaf | |
| CN105132172B (en) | A kind of method for preparing orris root flavonoids for tobacco from orris root | |
| CN104211690B (en) | Method for separating and purifying mangiferin from aquilaria sinensis leaves | |
| CN101348474A (en) | Method for preparing salvianolic acid B and tanshinol from Salvia miltiorrhiza stem | |
| CN101361786B (en) | Method for extracting and purifying total flavonoids from the forage cloverleaf | |
| CN102432420B (en) | Method for extracting and separating beta-elemene from Lantana camara | |
| CN105348333B (en) | A kind of chavicol disaccharide glycoside compound and its preparation method and application | |
| CN102432419A (en) | Method for extracting and separating beta-elemene from Eupatorium adenophorum | |
| CN112939742B (en) | Extraction and purification method of cannabinoid | |
| CN101735021B (en) | Method for separating and purifying magnolol and honokiol by HP-20 macroporous resin | |
| CN111848358A (en) | Method for preparing high-purity cannabidiol by combining macroporous resin enrichment with dynamic axial compression column system | |
| CN100564364C (en) | The method of Ligustrazine, forulic acid and volatile oil in a kind of comprehensively extracting and purifying Ligusticum wallichii | |
| CN101880301B (en) | Novel method for preparing paeoniflorin extract | |
| CN106699819B (en) | The preparation method of Penta-O-galloyl-D-glucopyranose chemical reference substance | |
| CN103951717B (en) | The method of benzoylpaeoniflorin and benzoyl lactone glucoside of Radix Paeoniae is prepared in a kind of extraction | |
| CN102293804B (en) | Optimized scheme for comprehensively utilizing silybum mariamum | |
| CN116410252A (en) | HSCCC preparation method of cynarin | |
| CN102391328B (en) | Method for simultaneously preparing chemical reference substances magnoloside A and magnoloside B | |
| CN112707878A (en) | Extraction method of phthalide compounds | |
| CN102391106A (en) | Method for extracting high-purity Adenostemmoic acid B from Adenostemma Lavenia | |
| CN104744528B (en) | The method for preparing parishin E and parishin G chemical reference substances simultaneously |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20201119 Address after: 214500, 6 Industrial Park, Xinqiao Town, Taizhou, Jiangsu, Jingjiang Patentee after: Jingjiang Huatai Machinery Manufacturing Co., Ltd Address before: 617000 Airport Road, Panzhihua, Sichuan, No. 10 Patentee before: PANZHIHUA University |
|
| TR01 | Transfer of patent right |