WO2003080045A1 - A process for isolating artemisinin from artemisia annua - Google Patents
A process for isolating artemisinin from artemisia annua Download PDFInfo
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- WO2003080045A1 WO2003080045A1 PCT/IB2002/001204 IB0201204W WO03080045A1 WO 2003080045 A1 WO2003080045 A1 WO 2003080045A1 IB 0201204 W IB0201204 W IB 0201204W WO 03080045 A1 WO03080045 A1 WO 03080045A1
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- WO
- WIPO (PCT)
- Prior art keywords
- artemisinin
- hexane
- extract
- aqueous
- extraction
- Prior art date
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- BLUAFEHZUWYNDE-NNWCWBAJSA-N artemisinin Chemical compound C([C@](OO1)(C)O2)C[C@H]3[C@H](C)CC[C@@H]4[C@@]31[C@@H]2OC(=O)[C@@H]4C BLUAFEHZUWYNDE-NNWCWBAJSA-N 0.000 title claims abstract description 91
- 229930101531 artemisinin Natural products 0.000 title claims abstract description 90
- 229960004191 artemisinin Drugs 0.000 title claims abstract description 89
- 238000000034 method Methods 0.000 title claims abstract description 70
- 240000000011 Artemisia annua Species 0.000 title claims description 26
- 235000001405 Artemisia annua Nutrition 0.000 title claims description 18
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims abstract description 202
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 92
- 239000000284 extract Substances 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 238000002425 crystallisation Methods 0.000 claims abstract description 24
- 230000008025 crystallization Effects 0.000 claims abstract description 24
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 48
- 238000000605 extraction Methods 0.000 claims description 46
- 239000007788 liquid Substances 0.000 claims description 35
- 239000002904 solvent Substances 0.000 claims description 34
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 32
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 26
- 239000010410 layer Substances 0.000 claims description 21
- 241000196324 Embryophyta Species 0.000 claims description 12
- 238000005325 percolation Methods 0.000 claims description 10
- 239000000469 ethanolic extract Substances 0.000 claims description 9
- 239000012044 organic layer Substances 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- 239000000126 substance Substances 0.000 claims description 7
- 239000004215 Carbon black (E152) Substances 0.000 claims description 6
- 229930195733 hydrocarbon Natural products 0.000 claims description 6
- 150000002430 hydrocarbons Chemical class 0.000 claims description 6
- 239000002798 polar solvent Substances 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 5
- 239000003125 aqueous solvent Substances 0.000 claims description 5
- 238000001704 evaporation Methods 0.000 claims description 5
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 claims description 4
- OFBQJSOFQDEBGM-UHFFFAOYSA-N Pentane Chemical compound CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 claims description 4
- 230000019612 pigmentation Effects 0.000 claims description 4
- 239000003610 charcoal Substances 0.000 claims description 3
- 230000008020 evaporation Effects 0.000 claims description 3
- 239000006096 absorbing agent Substances 0.000 claims description 2
- 239000006286 aqueous extract Substances 0.000 claims description 2
- 235000008216 herbs Nutrition 0.000 claims description 2
- -1 methylisobutyl Chemical group 0.000 claims description 2
- 238000005192 partition Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 238000000638 solvent extraction Methods 0.000 abstract description 23
- 238000002955 isolation Methods 0.000 abstract description 5
- 239000003430 antimalarial agent Substances 0.000 abstract description 3
- 235000003826 Artemisia Nutrition 0.000 abstract 1
- 240000006891 Artemisia vulgaris Species 0.000 abstract 1
- 235000003261 Artemisia vulgaris Nutrition 0.000 abstract 1
- 235000009052 artemisia Nutrition 0.000 abstract 1
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 39
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 14
- 239000000203 mixture Substances 0.000 description 14
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 12
- 239000013078 crystal Substances 0.000 description 10
- 238000004587 chromatography analysis Methods 0.000 description 9
- 238000012546 transfer Methods 0.000 description 7
- PLQMEXSCSAIXGB-SAXRGWBVSA-N (+)-artemisinic acid Chemical compound C1=C(C)CC[C@H]2[C@H](C)CC[C@@H](C(=C)C(O)=O)[C@H]21 PLQMEXSCSAIXGB-SAXRGWBVSA-N 0.000 description 6
- 239000002044 hexane fraction Substances 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 239000000741 silica gel Substances 0.000 description 5
- 229910002027 silica gel Inorganic materials 0.000 description 5
- 239000003208 petroleum Substances 0.000 description 4
- LZMOBPWDHUQTKL-RWMBFGLXSA-N artemisinic acid Natural products CC1=C[C@@H]2[C@@H](CCC[C@H]2C(=C)C(=O)O)CC1 LZMOBPWDHUQTKL-RWMBFGLXSA-N 0.000 description 3
- PLQMEXSCSAIXGB-UHFFFAOYSA-N artemisininic acid Natural products C1=C(C)CCC2C(C)CCC(C(=C)C(O)=O)C21 PLQMEXSCSAIXGB-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000004040 coloring Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 231100001261 hazardous Toxicity 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000002035 hexane extract Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- ZRSNZINYAWTAHE-UHFFFAOYSA-N p-methoxybenzaldehyde Chemical compound COC1=CC=C(C=O)C=C1 ZRSNZINYAWTAHE-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000003828 vacuum filtration Methods 0.000 description 2
- 239000000341 volatile oil Substances 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- 206010063094 Cerebral malaria Diseases 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- HATRDXDCPOXQJX-UHFFFAOYSA-N Thapsigargin Natural products CCCCCCCC(=O)OC1C(OC(O)C(=C/C)C)C(=C2C3OC(=O)C(C)(O)C3(O)C(CC(C)(OC(=O)C)C12)OC(=O)CCC)C HATRDXDCPOXQJX-UHFFFAOYSA-N 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 238000005267 amalgamation Methods 0.000 description 1
- 230000000078 anti-malarial effect Effects 0.000 description 1
- 229940033495 antimalarials Drugs 0.000 description 1
- 229960000981 artemether Drugs 0.000 description 1
- 229960002970 artemotil Drugs 0.000 description 1
- NLYNIRQVMRLPIQ-XQLAAWPRSA-N artemotil Chemical compound C1C[C@H]2[C@H](C)CC[C@H]3[C@@H](C)[C@@H](OCC)O[C@H]4[C@]32OO[C@@]1(C)O4 NLYNIRQVMRLPIQ-XQLAAWPRSA-N 0.000 description 1
- FIHJKUPKCHIPAT-AHIGJZGOSA-N artesunate Chemical compound C([C@](OO1)(C)O2)C[C@H]3[C@H](C)CC[C@@H]4[C@@]31[C@@H]2O[C@@H](OC(=O)CCC(O)=O)[C@@H]4C FIHJKUPKCHIPAT-AHIGJZGOSA-N 0.000 description 1
- 229960004991 artesunate Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 231100000481 chemical toxicant Toxicity 0.000 description 1
- 238000000658 coextraction Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- SXYIRMFQILZOAM-HVNFFKDJSA-N dihydroartemisinin methyl ether Chemical compound C1C[C@H]2[C@H](C)CC[C@H]3[C@@H](C)[C@@H](OC)O[C@H]4[C@]32OO[C@@]1(C)O4 SXYIRMFQILZOAM-HVNFFKDJSA-N 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000004210 ether based solvent Substances 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 201000004792 malaria Diseases 0.000 description 1
- 239000000401 methanolic extract Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 239000012454 non-polar solvent Substances 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 229930183339 qinghaosu Natural products 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 229930009674 sesquiterpene lactone Natural products 0.000 description 1
- 150000002107 sesquiterpene lactone derivatives Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/282—Artemisia, e.g. wormwood or sagebrush
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
- A61P33/06—Antimalarials
Definitions
- the present invention relates to a process for the isolation of artemisinin, an antimalarial agent from the herb of the Artemisia annua plant, comprising of extracting the herb with ethanol, partitioning of the extract between water and hexane, followed by evaporative crystallization of artemisinin from hexane phase to produce substantially pure artemisinin.
- BACKGROUND ART Artemisinin (Qinghaosu) is a sesquiterpene lactone endoperoxide having potent antimalarial activity (Klayman DL, 1985, Science 228, 1049).
- Another method of artemisinin extraction involves the extraction of plant material with hexane, followed by partitioning the extract between hexane and acetonitrile followed by chromatographing the acetonitrile phase on silica gel (ElFerali FS, ElSohli HN, 1990, US Patent No. 4,952,603). Slow extraction of artemisnin in hexane needs hot percolation for longer duration, while for partitioning step, acetonitrile being relatively costly relatively costly solvent, increases cost of production for artemisinin.
- Another disadvantage of this method is that for obtaining artemisinin column chromatography is unavoidable.
- artemisinic acid being predominant, it tends to elute with artemisinin, thus affecting the purity of artemisinin.
- the main object of the present invention is to provide a simple, rapid, cost effective and practical method for the isolation of artemisinin from plant Artemisia annua without using chromatography, which yields artemisinin in substantial quantities and purity obviating the drawbacks of the prior art.
- the present invention provides a process for the preparation of artemisinin from Artemisia annua, said process comprising:
- step (ii) concentrating the extract of step (i) to recover the polar solvent; (iii) adding water to the aforesaid concentrated extract;
- step(iv) adding hydrocarbon solvent to partition the aqueous extract of step(iii) into an aqueous layer and an organic layer;
- step (v) separating the organic layer from the solution of step (iv);
- step (vi) concentrating the organic layer of step (v) to obtain oily liquid; (vii) adding ethyl acetate to the oily liquid of step (vi);
- step (viii) treating the solution of step (vii) with a conventional color absorbing substance to remove greenish pigmentation;
- step (x) evaporating the solution followed by crystallization to obtain pure artemisinin.
- step (i) dry leaves are taken for extraction.
- the leaves are powdered before extraction.
- the non-aqueous polar solvent is selected from the group consisting of ethanol, methanol, acetone, methylisobutyl and hexane.
- the non-aqueous solvent is ethanol.
- the ratio of the plant parts to the non- aqueous solvent is in the range of 1:5 to 1:10.
- the extraction of the plant parts is effected by percolating the herbs with ethanol for four to six hours.
- percolation of the herb is effected at temperatures between 20 to 50°C.
- step (ii) wherein in step (ii) the volume of the ethanolic extract is reduced to 1/20* to l/lOO* of its original volume.
- step (ii) wherein in step (ii) the solvent recovered after concentration is reused in the process step (i).
- step (iii) water is added four times the quantity of the concentrated ethanolic extract.
- step (iv) the hydrocarbon solvent used is selected from the group consisting of pentane, hexane and heptane.
- step (iv) wherein in step (iv) the hydrocarbon solvent used is hexane.
- step (iv) wherein in step (iv) the aqueous ethanolic extract is partioned in hexane in the ratio of 1 : 1 or 2: 1.
- step (v) wherein in step (v) the organic layer is concentrated l/20 th to 1/100* of its original volume.
- step (vii) 10-20% (v/v) ethyl acetate is added to the concentrated hexane solution to induce crystallization of artemisinin from the liquid.
- the color-absorbing agent is selected from the group consisting of celite, activated charcoal and charcoal.
- step (x) artemisinin is obtained by slow evaporation followed by crystallization.
- the process developed in the present invention provides an improved method of isolating the artemisinin from Artemisia annua plant populations.
- Our method involves cold extraction with ethanol/methanol, which on one hand allows complete extraction of artemisinin with rapidity and saves energy on the other hand in contrast to the methods of extraction using hexane or petroleum ether practiced routinely. Extraction in hexane or ether was very slow and needed hot percolation over several hours.
- Partitioning of the extract between water and hexane is economical and a completely non-obvious approach to enrich the extract with artemisinin, as 60-70% impurities of extract are left in water layer and complete transfer of artemisinin in to hexane layer.
- the hexane fraction thus obtained can be used for direct crystallization after a step of de-colorization for obtaining artemisinin without chromatography.
- invention gives a new approach of isolating the artemisinin from its natural source Artemisia annua plant populations without using chromatography.
- the mild conditions used for extraction give a clean plant extract and avoid the possible degradation of artemisinin.
- dried pulverized leaves of Artemisia annua may be extracted by continuous percolation over a period of four to six hours using five to ten fold volume of the non-aqueous solvent selected from ethanol or methanol.
- the said extraction process may be repeated three to five times using same solvent ratios to ensure maximum extraction of artemisinin from the herb.
- the resulting extract can be concentrated to 1 to 5% of the original volume by distillation under vacuum.
- the recovered solvent may be used again in the extraction process.
- For partitioning the aqueous content and hexane may be used in a ratio of 1 : 1 or 2 : 1 v/v. Partitioning of aqueous content with non-polar solvent may be repeated three to five times using the same solvent ratio in order to ensure maximum transfer of artemisinin to hexane fraction.
- the combined hexane fractions may be pooled together before they are distilled under vacuum (to recover the solvent for using again) to obtain 1-5% of its original volume.
- the concentrated liquid may be a light to dark green oily liquid.
- Ethyl acetate (10- 20% v/v) is added to it. To remove the green pigmentation this liquid is treated with 1- 3%w/v of any colour absorbing substance activated charcoal. The yellowish liquid obtained after removal of activate charcoal (by filtration) may be subjected to the evaporative crystallization yielding substantially pure artemisinin without using chromatography.
- the extraction of dry, pulverized leaves with non-aqueous ethanol may have several advantages over the previously used hexane or di- ethyl ether solvents. Besides extraction being rapid, ethanol will extract less amount of fatty material, which are considered as an obstacle in the purification and crystallization steps.
- the extract is partitioned between water and hexane in order to enrich the extract with respect to the artemsinin and remove as many impurities as possible from the concentrate to facilitate the crystallization of artemisinin.
- partitioning of the ethanol extract is carried out using water and hexane in a ratio ranging from (1 : 1 or 2 : 1), the process of partitioning can be repeated 3 - 5 times to ensure maximum transfer of artemisinin to hexane layer.
- the partitioning step results in a substantially exclusive transfer of the artemisinin into the hexane layer with concomitant reduction in the amount of material, i.e. only 30 to 35% weight of the original extract is transferred in to hexane layer.
- hexane fraction can directly be used for crystallization by slow evaporation after concentrating the hexane fraction and mixing 10-20% (v/v) of ethyl acetate.
- the TLC was carried out in a glass TLC tank saturated with the mobile phase hexane: diethyl ether (1 : 1), plates were developed to a height of 15 cm. Later plates were dried and spots were visualized by immersing the plates in a developing reagent (glacial acetic acid: sulfuric acid: anisaldehyde, 50: 1 : 0.5), followed by heating the plate at 110°C for 15 minutes or until pink spots of artemisinin appeared. For quantification, TLC spots corresponding to artemisinin were scanned at 540 and 610 nm dual wavelength mode of TLC scanner. The results obtained are presented in the form of % artemisinin extracted in to the solvent and are summarized in table - 1.
- Table 2 Data representing the trend of partitioning of artemisinin between hexane and ethanolic solution with water.
- artemisisnin can be selectively transferred from ethatnolic extract of Artemisia annua in to hexane by maintaining the above ratios. From hexane fraction, artemisinin can be crystallized directly using known methods of evaporative crystallization.
- Dry pulverized leaves of Artemisia annua (100 g) was extracted by continuous percolation over a period of four hours using 600 ml of ethanol at 30°C temperature. The process of extraction was repeated four times using same solvent ratios.
- the combined extract was concentrated under vacuum to reduce the volume to 50 ml, volume was made up to 250 ml by adding water to it. The resulting mixture was partitioned with 250-ml hexane four times.
- the combined hexane layer was reduced under vacuum (to recover hexane) to 5% of its original volume to result in dark green oily liquid.
- a 20% v/v ethyl acetate was mixed to this liquid followed by treating this with 1% w/v activated charcoal.
- Dry pulverized leaves of Artemisia annua (100 g) was extracted by continuous percolation over a period of four hours using 1.0 litre of ethanol at 30°C temperature. The process of extraction was repeated four times using same solvent ratios.
- the combined extract was concentrated under vacuum to reduce the volume to 80 ml, volume was made up to 400 ml by adding water to it.
- the resulting mixture was partitioned with 400-ml hexane four times.
- the combined hexane layer was reduced under vacuum to 50 ml volume to result in dark green oily liquid.
- a 20% v/v ethyl acetate was mixed to this liquid followed by treating this with 1% w/v activated charcoal.
- the liquid was filtered as in example 3 and subjected to evaporative crystallization.
- the artemisinin crystals obtained were weighed to be 605 mg.
- EXAMPLE 7 Dried pulverized leaves of Artemisia annua (1.0 kg) were extracted by continuous percolation over a period of four hours using 6 litres of ethanol at 30°C temperature. The process of extraction was repeated four times using same solvent ratios. The combined extract was concentrated under vacuum to reduce the volume to 400 ml, volume was made up to 2.0 liters by adding water to it. The resulting mixture was partitioned with 2.0 liters hexane three times.
- Dried pulverized leaves of Artemisia annua (100 g) were extracted by continuous percolation over a period of four hours using 600 ml of methanol at 30°C temperature. The process of extraction was repeated four times using same solvent ratios.
- the combined extract was concentrated under vacuum to reduce the volume to 50 ml, volume was made up to 250 ml by adding water to it.
- the resulting mixture was partitioned with 250-ml hexane three times.
- the combined hexane layer was reduced 5% of its original volume to result in dark green oily liquid.
- a 20% v/v ethyl acetate was mixed to this liquid followed by treating this with 1% w/v activated charcoal.
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- General Chemical & Material Sciences (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
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Priority Applications (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2003577873A JP4339699B2 (ja) | 2002-03-25 | 2002-03-25 | アーテミシア・アニュア(Artemisia annua)からアーテミシニン(artemisinin)を単離する方法 |
CNA028290003A CN1627942A (zh) | 2002-03-25 | 2002-03-25 | 黄花蒿离析苦艾素的方法 |
PCT/IB2002/001204 WO2003080045A1 (en) | 2002-03-25 | 2002-03-25 | A process for isolating artemisinin from artemisia annua |
AU2002253443A AU2002253443A1 (en) | 2002-03-25 | 2002-03-25 | A process for isolating artemisinin from artemisia annua |
US10/112,070 US6685972B2 (en) | 2002-03-25 | 2002-03-27 | Process for isolating artemisinin from Artemisia annua |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/IB2002/001204 WO2003080045A1 (en) | 2002-03-25 | 2002-03-25 | A process for isolating artemisinin from artemisia annua |
US10/112,070 US6685972B2 (en) | 2002-03-25 | 2002-03-27 | Process for isolating artemisinin from Artemisia annua |
Publications (1)
Publication Number | Publication Date |
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WO2003080045A1 true WO2003080045A1 (en) | 2003-10-02 |
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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PCT/IB2002/001204 WO2003080045A1 (en) | 2002-03-25 | 2002-03-25 | A process for isolating artemisinin from artemisia annua |
Country Status (4)
Country | Link |
---|---|
JP (1) | JP4339699B2 (zh) |
CN (1) | CN1627942A (zh) |
AU (1) | AU2002253443A1 (zh) |
WO (1) | WO2003080045A1 (zh) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008502712A (ja) * | 2004-06-11 | 2008-01-31 | ユニジェン インク. | 竹又は竹抽出物を含有してなるアンドロゲン作用組成物 |
CN100445726C (zh) * | 2006-08-09 | 2008-12-24 | 重庆医科大学 | 一种青蒿中青蒿素含量的测定方法 |
CN103013655A (zh) * | 2013-01-09 | 2013-04-03 | 新疆大学 | 苦艾挥发油的制备方法 |
CN111141846A (zh) * | 2019-12-31 | 2020-05-12 | 广西仙草堂制药有限责任公司 | 一种青蒿中青蒿素的含量测定方法 |
CN114181226A (zh) * | 2021-11-04 | 2022-03-15 | 河南天源药物研究有限公司 | 一种青蒿精油的提纯制备工艺 |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102890087A (zh) * | 2012-11-13 | 2013-01-23 | 宁夏医科大学 | 一种黄花蒿等中药材及含青蒿素成分样品中青蒿素含量的测定方法 |
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EP0908460A1 (en) * | 1997-10-06 | 1999-04-14 | Council of Scientific and Industrial Research | A process for the simultaneous production of artemisinin and essential oil from the plant artemisia annua |
US6197767B1 (en) * | 1997-04-26 | 2001-03-06 | Paracure, Inc. | Arglabin compounds and therapeutic uses thereof |
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2002
- 2002-03-25 WO PCT/IB2002/001204 patent/WO2003080045A1/en active Application Filing
- 2002-03-25 JP JP2003577873A patent/JP4339699B2/ja not_active Expired - Fee Related
- 2002-03-25 AU AU2002253443A patent/AU2002253443A1/en not_active Abandoned
- 2002-03-25 CN CNA028290003A patent/CN1627942A/zh active Pending
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US6197767B1 (en) * | 1997-04-26 | 2001-03-06 | Paracure, Inc. | Arglabin compounds and therapeutic uses thereof |
EP0908460A1 (en) * | 1997-10-06 | 1999-04-14 | Council of Scientific and Industrial Research | A process for the simultaneous production of artemisinin and essential oil from the plant artemisia annua |
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008502712A (ja) * | 2004-06-11 | 2008-01-31 | ユニジェン インク. | 竹又は竹抽出物を含有してなるアンドロゲン作用組成物 |
CN100445726C (zh) * | 2006-08-09 | 2008-12-24 | 重庆医科大学 | 一种青蒿中青蒿素含量的测定方法 |
CN103013655A (zh) * | 2013-01-09 | 2013-04-03 | 新疆大学 | 苦艾挥发油的制备方法 |
CN111141846A (zh) * | 2019-12-31 | 2020-05-12 | 广西仙草堂制药有限责任公司 | 一种青蒿中青蒿素的含量测定方法 |
CN114181226A (zh) * | 2021-11-04 | 2022-03-15 | 河南天源药物研究有限公司 | 一种青蒿精油的提纯制备工艺 |
Also Published As
Publication number | Publication date |
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JP4339699B2 (ja) | 2009-10-07 |
AU2002253443A1 (en) | 2003-10-08 |
CN1627942A (zh) | 2005-06-15 |
JP2005526096A (ja) | 2005-09-02 |
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