CN102603764A - Extraction method of columbin - Google Patents
Extraction method of columbin Download PDFInfo
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- CN102603764A CN102603764A CN2011100262091A CN201110026209A CN102603764A CN 102603764 A CN102603764 A CN 102603764A CN 2011100262091 A CN2011100262091 A CN 2011100262091A CN 201110026209 A CN201110026209 A CN 201110026209A CN 102603764 A CN102603764 A CN 102603764A
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- columbin
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
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Abstract
The invention discloses an extraction method of columbin. The extraction method disclosed by the invention comprises the following steps of: applying a technology combining supercritical carbon dioxide extraction with macroporous resin separation and purification, taking acetone, ethanol or methanol as an entrainer, and taking CO2 as a medium to extract an active ingredient, namely the columbin, from tinospora root; and performing macroporous resin chromatography, recrystallization and other steps on an extract to get a high-purity extract. Compared with the existing organic solvent extraction method, the extraction method has the advantages of simple process, short time, high purity, relatively light environmental pollution and the like.
Description
Technical field:
The invention belongs to Chinese medicine extracts active ingredients stripping technique, relate to a kind of process for extracting of Columbin.
Background technology:
Root of Hairystalk Tinospora is the dried root of menispermaceous plants limacia sagittata Tinospara sagittata (Oliv.) Gagncp. or Root of Hairystalk Tinospora Tinosporacapillipes Gagnep..Limacia sagittata is distributed in Shaanxi, each province, the Yangtze valley to Guangdong and Guangxi Provinces, each provinces and regions, southwest.Root of Hairystalk Tinospora is distributed in provinces and regions such as Hunan, Hubei, Guangxi, Guangdong, Sichuan, Guizhou, Yunnan.The piece root contains diterpenes bitter principle Columbin (tinosporine), palmatine, contains the furanoditerpene glycosides again---tinoside (leurocolombine 4-β-D-glucoside).05 edition " pharmacopeia " is to be quality index with the palmatine hydrochloride, and the content of palmatine hydrochloride in Root of Hairystalk Tinospora is less, and is not the characteristic chemical constituent of Root of Hairystalk Tinospora, and therefore version pharmacopeia in 2010 Columbin that content is higher is as the index property composition of Root of Hairystalk Tinospora.
Columbin (Columbin) has another name called tinosporine, columbin, Columbin, is from Root of Hairystalk Tinospora, to separate the diterpenes composition that obtains.Molecular formula C
20H
22O
6, molecular weight 358.38, extremely bitter needle crystal (methyl alcohol), mp.195 ℃.Water-soluble hardly, 1g is dissolved in 30ml acetone, 50ml ETHYLE ACETATE, 75ml methyl alcohol, is dissolved in chloroform and methylene dichloride.Pharmacological research shows: Columbin has significant anti-inflammatory activity, and presents dose-effect relationship: can come inhibited oxidation azomethane inductive rat colon cancer through the cell regeneration that suppresses in the colon vesica significantly; The hydrolytic action that can suppress the catalytic erythrocyte shell of cobra venom acid phosphorus lipase A2; Can suppress the absorption of SUV in the trypanosoma bocagei blood flow; Can influence liver medicine metabolic enzyme regulated by ciclobarbital soluble or urethane and Chloralosane mixture inductive rat length of one's sleep.Oral Columbin can dose-dependent reduction liver-7-alkoxycoumarin-O-dealkylas (ACD), and ACD has reflected the activity of P450.Appoint equality also to find that through experiment columbin has enzymatic kinetics metabolic characteristic.At present, the extractive technique of Columbin mainly is that ether Suo Shi extracts, and this method products obtained therefrom purity is low, and solvent toxicity is bigger.Li Hongbo adopts methanol eddy extraction, SX, and normal pressure, pressurization, positive and reversed-phase silica gel column chromatography separate repeatedly; Shi Qirong adopt 95% alcohol reflux, chloroform and n-butanol extraction, repeatedly silica gel column chromatography (dry method), ODS-C18 column chromatography, partly prepare purification process such as liquid phase, PharmadexLH-20 isochromatic spectrum technology, recrystallization and obtain Columbin.Above-mentioned two kinds of technologies exist extraction yield low, and complex process, consumption of organic solvent is big, environmental pollution is serious, deficiency such as with short production cycle.
Summary of the invention:
The object of the present invention is to provide a kind of process for extracting of Columbin, adopt supercritical CO
2Steps such as extraction, macroporous resin column chromatography, recrystallization, the extraction separation purifying obtains the Columbin of higher degree from Root of Hairystalk Tinospora, and the Columbin purity that this method is extracted is high, low in the pollution of the environment, yield is high, cost is low, but enlargement of scale production.
The present invention realizes through following technical scheme:
A kind of process for extracting of Columbin is characterized in that Root of Hairystalk Tinospora piece root is ground into meal, puts into supercritical CO
2In the extraction kettle, add an amount of organic solvent again, feed CO as entrainment agent
2As extracting medium, at extracting pressure 15-30MPa, extract under the temperature 30-45 ℃ of condition, after extraction in 1-4 hour, resolve; Resolve pressure 5-10MPa, temperature 50-60 ℃, collect extract, extract is joined in the macroporous adsorptive resins; Flow velocity 1.0-2.0BV/h with the washing of 2-5 times of column volume, discards water lotion earlier; With the 20-30% ethanol elution of 3-6 times of column volume, use the 40-100% ethanol elution at last again, collect the 40-100% ethanol eluate; Be concentrated into small volume, crystallization, ethyl alcohol recrystallization.
Said supercritical CO
2The optional volume percent of entrainment agent is acetone, the methanol solution of 70-100% in the extraction step, and add-on is the 5-20% of medicinal material amount.
Said supercritical CO
2CO in the extraction step
2Flow velocity is 15-30L/h.
A kind of among the optional ADS-7 of said macroporous adsorbent resin, ADS-8, D101, AB-8, the HPD100.
The present invention and prior art compare, and extraction efficiency is high, product purity is high, environmental pollution is less, and preparation amount is big, but enlargement of scale production.
Embodiment:
To combine specific embodiment that the present invention is described further below, the following example is intended to describe for example the present invention, rather than limits the present invention by any way.
Embodiment 1:
Root of Hairystalk Tinospora piece root herb is ground into 20 order meal, gets 2kg and drop in the supercritical extraction jar, add volume percent and be 80% acetone soln as entrainment agent, add-on is 0.12kg (raw material weight 6%); At extracting pressure 18.5MPa, 32 ℃ of extraction temperature, CO
2Extraction is 1 hour under the condition of flow 18L/h; The pressure that control is resolved jar is 5.4MPa, 56 ℃ of temperature, separate extract; With 60 ℃ of hot water dissolving's extracts of 1 times of amount, last ADS-7 macroporous adsorptive resins, absorption flow velocity 1.0-2.0BV/h with the washing of 2 times of column volumes, discards water lotion earlier; Use 4 times of column volume 50% ethanol elutions again, use 90% ethanol elution at last, collect 90% ethanol eluate, above-mentioned elutriant is evaporated to small volume, crystallization is used ethyl alcohol recrystallization 2 times again, and lyophilize gets product 20.8g.Analyze through HPLC, Columbin purity is 98.6%.
Embodiment 2:
Root of Hairystalk Tinospora piece root herb is ground into 20 order meal, gets 5kg and drop in the supercritical extraction jar, add volume percent and be 90% methanol solution as entrainment agent, add-on is 0.12kg (raw material weight 10%); At extracting pressure 22.3MPa, 45 ℃ of extraction temperature, CO
2Extraction is 3 hours under the condition of flow 15L/h: the pressure that control is resolved jar is 8.7MPa, 50 ℃ of temperature, separate extract; With 60 ℃ of hot water dissolving's extracts of 1 times of amount, last D101 macroporous adsorptive resins, absorption flow velocity 1.0-2.0BV/h with the washing of 3 times of column volumes, discards water lotion earlier; Use 6 times of column volume 50% ethanol elutions again, use 90% ethanol elution at last, collect 90% ethanol eluate, above-mentioned elutriant is evaporated to small volume, crystallization is used ethyl alcohol recrystallization 2 times again, and lyophilize gets product 51.2g.Analyze through HPLC, Columbin purity is 98.0%.
Embodiment 3:
Root of Hairystalk Tinospora piece root herb is ground into 20 order meal, gets 5kg and drop in the supercritical extraction jar, the acetone soln that adds volume percent 100% is as entrainment agent, and add-on is 0.12kg (raw material weight 12%); At extracting pressure 27MPa, 35 ℃ of extraction temperature, CO
2Extraction is 2 hours under the condition of flow 23L/h; The pressure that control is resolved jar is 6.3MPa, 52 ℃ of temperature, separate extract: with 60 ℃ of hot water dissolving's extracts of 1 times of amount, last AB-8 macroporous adsorptive resins adsorbs flow velocity 1.0-2.0BV/h, and elder generation discards water lotion with the washing of 5 times of column volumes; Use 3 times of column volume 50% ethanol elutions again, use 90% ethanol elution at last, collect 90% ethanol eluate, above-mentioned elutriant is evaporated to small volume, crystallization is used ethyl alcohol recrystallization 2 times again, and lyophilize gets product 48.6g.Analyze through HPLC, Columbin purity is 98.4%.
Embodiment 4:
Root of Hairystalk Tinospora piece root herb is ground into 20 order meal, gets 20kg and drop in the supercritical extraction jar, the acetone soln that adds volume percent 70% is as entrainment agent, and add-on is 0.12kg (raw material weight 18%); At extracting pressure 15MPa, 43 ℃ of extraction temperature, CO
2Extraction is 4 hours under the condition of flow 27L/h: the pressure that control is resolved jar is 5.0MPa, 60 ℃ of temperature, separate extract; With 60 ℃ of hot water dissolving's extracts of 1 times of amount, last ADS-8 macroporous adsorptive resins, absorption flow velocity 1.0-2.0BV/h with the washing of 6 times of column volumes, discards water lotion earlier; Use 5 times of column volume 50% ethanol elutions again, use 90% ethanol elution at last, collect 90% ethanol eluate, above-mentioned elutriant is evaporated to small volume, crystallization is used ethyl alcohol recrystallization 2 times again, and lyophilize gets product 205g.Analyze through HPLC, Columbin purity is 99.1%.
Embodiment 5:
Root of Hairystalk Tinospora piece root herb is ground into 20 order meal, gets 20kg and drop in the supercritical extraction jar, the methanol solution that adds volume percent 80% is as entrainment agent, and add-on is 0.12kg (raw material weight 20%); At extracting pressure 30.5MPa, 38 ℃ of extraction temperature, CO
2Extraction is 3 hours under the condition of flow 20L/h; The pressure that control is resolved jar is 10MPa, 54 ℃ of temperature, separate extract: with 60 ℃ of hot water dissolving's extracts of 1 times of amount, last D101 macroporous adsorptive resins adsorbs flow velocity 1.0-2.0BV/h, and elder generation discards water lotion with the washing of 3 times of column volumes; Use 4 times of column volume 50% ethanol elutions again, use 90% ethanol elution at last, collect 90% ethanol eluate, above-mentioned elutriant is evaporated to small volume, crystallization is used ethyl alcohol recrystallization 2 times again, and lyophilize gets product 198.5g.Analyze through HPLC, Columbin purity is 98.8%.
Claims (4)
1. the process for extracting of a Columbin is characterized in that Root of Hairystalk Tinospora piece root is ground into meal, puts into supercritical CO
2In the extraction kettle, add an amount of organic solvent again, feed CO as entrainment agent
2As extracting medium, at extracting pressure 15-30MPa, extract under the temperature 30-45 ℃ of condition, after extraction in 1-4 hour, resolve; Resolve pressure 5-10MPa, temperature 50-60 ℃, collect extract, extract is joined in the macroporous adsorptive resins; Flow velocity 1.0-2.0BV/h with the washing of 2-5 times of column volume, discards water lotion earlier; With the 20-30% ethanol elution of 3-6 times of column volume, use the 40-100% ethanol elution at last again, collect the 40-100% ethanol eluate; Be concentrated into small volume, crystallization, ethyl alcohol recrystallization.
2. according to the process for extracting of the said Columbin of claim 1, it is characterized in that said supercritical CO
2The optional volume percent of entrainment agent is acetone, methyl alcohol, the ethanolic soln of 70-100% in the extraction step, and add-on is the 5-20% of medicinal material amount.
3. according to the process for extracting of the said Columbin of claim 1, it is characterized in that said supercritical CO
2CO in the extraction step
2Flow velocity is 15-30L/h.
4. according to the process for extracting of the said Columbin of claim 1, it is characterized in that a kind of among the optional ADS-7 of said macroporous adsorbent resin, ADS-8, D101, AB-8, the HPD100.
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| CN2011100262091A CN102603764A (en) | 2011-01-25 | 2011-01-25 | Extraction method of columbin |
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| CN2011100262091A CN102603764A (en) | 2011-01-25 | 2011-01-25 | Extraction method of columbin |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105330672A (en) * | 2015-12-22 | 2016-02-17 | 姚蓉 | Method for extracting high purity furanodiene and palmatine from radix tinosporae |
| CN110269885A (en) * | 2019-06-06 | 2019-09-24 | 中国人民解放军南部战区总医院 | Radix Tinosporae extract is preparing the application in overriding resistance Acinetobacter bauamnnii drug |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH072682A (en) * | 1993-06-17 | 1995-01-06 | Iatron Lab Inc | Central nervous system activation agent |
| CN1839825A (en) * | 2006-01-19 | 2006-10-04 | 中国人民解放军第二军医大学 | Use of columbin, isocolumbin, deoxidized fibraurin in preparing antiphlogistic, stomach convulsion, stomach ache medicine |
| CN1947715A (en) * | 2006-09-07 | 2007-04-18 | 中国人民解放军第二军医大学 | Application of Gulunbin, iso-gulunbin and fibleucin for preparing medicine for treating arthritis |
-
2011
- 2011-01-25 CN CN2011100262091A patent/CN102603764A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH072682A (en) * | 1993-06-17 | 1995-01-06 | Iatron Lab Inc | Central nervous system activation agent |
| CN1839825A (en) * | 2006-01-19 | 2006-10-04 | 中国人民解放军第二军医大学 | Use of columbin, isocolumbin, deoxidized fibraurin in preparing antiphlogistic, stomach convulsion, stomach ache medicine |
| CN1947715A (en) * | 2006-09-07 | 2007-04-18 | 中国人民解放军第二军医大学 | Application of Gulunbin, iso-gulunbin and fibleucin for preparing medicine for treating arthritis |
Non-Patent Citations (4)
| Title |
|---|
| 《卫生职业教育》 20020430 袁崇均，等 大孔树脂-比色法测定金果榄中总内酯的含量 第94-95页 1-4 第20卷, 第4期 * |
| LIN-MEI SHI，等: "Two New Furanoid Diterpenoids from Tinospora sagittata", 《HELVETICA CHIMICA ACTA》 * |
| 姚健，等: "甘肃黄花蒿超临界CO2 萃取产物化学成分的差异性分析", 《草业科学》 * |
| 袁崇均，等: "大孔树脂—比色法测定金果榄中总内酯的含量", 《卫生职业教育》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105330672A (en) * | 2015-12-22 | 2016-02-17 | 姚蓉 | Method for extracting high purity furanodiene and palmatine from radix tinosporae |
| CN110269885A (en) * | 2019-06-06 | 2019-09-24 | 中国人民解放军南部战区总医院 | Radix Tinosporae extract is preparing the application in overriding resistance Acinetobacter bauamnnii drug |
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Application publication date: 20120725 |