CN103183658A - Method for extracting effective component of coleus forskohlii - Google Patents

Method for extracting effective component of coleus forskohlii Download PDF

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CN103183658A
CN103183658A CN2011104588184A CN201110458818A CN103183658A CN 103183658 A CN103183658 A CN 103183658A CN 2011104588184 A CN2011104588184 A CN 2011104588184A CN 201110458818 A CN201110458818 A CN 201110458818A CN 103183658 A CN103183658 A CN 103183658A
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macroporous resin
ethanol
ods
extract
column chromatography
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黄建明
田璐璐
汪亚勤
殷嘉珺
刘真君
谢道涛
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Fudan University
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Fudan University
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Abstract

本发明属中药制药领域，涉及一种提取毛喉鞘蕊花有效成分的方法。具体是一种提取毛喉鞘蕊花有效成分异佛司可林的方法。本发明将毛喉鞘蕊花全草或根粉碎后，以含水乙醇提取，大孔树脂柱层析、十八烷基键合硅胶柱层析、重结晶等方法进行纯化，制得纯度大于90％的异佛司可林。本方法制备工艺环保，易放大生产，制得的异佛司可林纯度可高达90％甚至98％，可用作药品开发的原料和保健品的功能添加剂。The invention belongs to the field of traditional Chinese medicine pharmacy, and relates to a method for extracting effective components of Coleus forskohlii. Specifically, it is a method for extracting isofeskolin, an active ingredient of Coleus forskohlii. In the present invention, the whole herb or root of Coleus forskohlii is crushed, extracted with aqueous ethanol, purified by macroporous resin column chromatography, octadecyl bonded silica gel column chromatography, recrystallization and other methods, and the obtained product has a purity greater than 90 % Isoferskolin. The preparation process of the method is environmentally friendly, easy to scale up production, and the purity of the prepared isofaskolin can be as high as 90% or even 98%, and can be used as raw materials for drug development and functional additives for health care products.

Description

A kind of method of extracting Coleus forskohlii Briq. effective constituent
Technical field
The invention belongs to field of traditional Chinese medicine pharmacy, relate to a kind of method of extracting Coleus forskohlii Briq. effective constituent.Concrete a kind of method of extracting Coleus forskohlii Briq. effective constituent Isoforskolin.
Background technology
Coleus forskohlii Briq. Coleus forskohlii (Willd) Briq. is Labiatae Labiatae Coleus Coleus Lour. plant, can be used for treating diseases such as cardiovascular disorder, flu, cough and respiratory disorder.Isoforskolin (isoforskolin) is the main effective constituent of homemade Coleus forskohlii Briq., effects such as prior art discloses it and has step-down, reduces intraocular pressure, relievings asthma, platelet aggregation-against, anti-injury of lung.This compound has multiple biological activity, and the application prospect aspect medicine and protective foods exploitation is good.
The correlation technique of present existing extraction Isoforskolin mainly contains: (1) adopts aqueous ethanol to extract, organic solvent extraction and technology such as silica gel column chromatography separates prepare different Buddhist department crin, there is following defective in this kind technology: use poisonous organic solvents such as a large amount of chloroforms, methylene dichloride, sherwood oil or acetone, and be difficult for regeneration reusable parting material silica gel (Chinese patent publication number CN 1539493A and CN 101259123A); (2) supercritical fluid extraction, this method equipment complexity, investment is big, also unexposed this method make the purity (Chinese patent publication number CN 1539493A) of Isoforskolin; (3) adopt aqueous ethanol to extract, macroporous resin purification, this method is easy, be suitable for big production, but the content of Isoforskolin only is 8% and 20% (Chinese patent ZL200410084646.9) in extract obtained.Therefore, about causing this area investigator's concern as the method that can extract the high purity Isoforskolin.
Summary of the invention:
The objective of the invention is to overcome the defective of prior art, a kind of method of extracting Coleus forskohlii Briq. effective constituent is provided, especially a kind of method of extracting Coleus forskohlii Briq. effective constituent Isoforskolin.
The present invention extracts with aqueous ethanol after Coleus forskohlii Briq. herb or root are pulverized, and methods such as macroporous resin column chromatography, octadecyl silane column chromatography, recrystallization are carried out purifying, makes purity greater than 90% Isoforskolin.Present method preparation technology's environmental protection is easily amplified and is produced, and the Isoforskolin purity that makes can be used as the raw material of drug development and the functional additive of healthcare products up to 90% even 98%.
Particularly, the method for extraction Coleus forskohlii Briq. effective constituent Isoforskolin of the present invention is characterized in that it comprises step:
(1) extracts: after getting the pulverizing of Coleus forskohlii Briq. herb or root, with 70～95% extraction using alcohols, concentrated extracting solution, concentrated solution direct filtration, or add flocculation agent after-filtration, filtrate for later use;
(2) macroporous resin column chromatography purifying: above-mentioned filtrate is regulated pH value back and is passed through macroporous resin column, and water and concentration are 20～40% ethanol drip washing successively, use 60～85% ethanol elutions again, elutriant is concentrated into dried, gets macroporous resin extract;
(3) octadecyl silane (ODS) column chromatography purification: get above-mentioned macroporous resin extract, with etc. weight ODS mix, or with behind a spot of as far as possible dissolve with ethanol, last ODS column purification, with 20～35% ethanol drip washing impurity, use 40～65% ethanol elutions again, elutriant is concentrated into dried, get the ODS extract;
(4) recrystallization: above-mentioned ODS extract carries out recrystallization with the mixed solution of ethyl acetate, acetone, ethanol or described solvent, separate out white crystal after, leach crystallization, drying gets purity greater than 90% Isoforskolin;
(5) preparation liquid phase purifying: the Isoforskolin crystal that makes with dissolve with methanol after, be further purified through the preparation liquid phase, get purity greater than 98% Isoforskolin.
Among the present invention, the Isoforskolin purity that makes is counted by weight percentage.
Among the present invention, the polystyrene type porousness polymeric adsorbent of the preferred nonpolar or low-pole of described macroporous resin.
In the step 2 of the present invention, the pH value of filtrate scope before the described upward macroporous resin is 5～10.
In the step 2 of the present invention, the macroporous resin column chromatography purifying carries out 1～3 time.
In the step 3 of the present invention, described ODS column chromatography purification carries out 1～3 time.
In the step 4 of the present invention, recrystallization 2～3 times repeatedly.
Among the present invention, adopt the high performance liquid chromatography evaporative light-scattering detector to measure the purity of Isoforskolin, the purity of calculating Isoforskolin by the peak area normalization method is more than 90%.
The present invention has the technology environmental protection compared with the prior art, is suitable for suitability for industrialized production, and the Isoforskolin purity advantages of higher that makes is embodied as:
(1) solvent of Shi Yonging is mainly the second alcohol and water, recyclable utilization, environmentally safe;
(2) the column chromatography for separation material adopts macroporous resin and octadecyl silane, uses repeatedly after renewable;
(3) the Isoforskolin purity height that makes can reach more than 90%.
For the ease of understanding, below will describe in detail by the method for specific embodiment to extraction Coleus forskohlii Briq. effective constituent of the present invention.It needs to be noted, specific examples only is in order to illustrate, obviously those of ordinary skill in the art can illustrate according to this paper, within the scope of the invention the present invention is made various corrections and change, and these corrections and change are also included in the scope of the present invention.
Embodiment:
Embodiment 1
(1) the Coleus forskohlii Briq. herb is 5 kilograms, pulverizes, and crosses 20 mesh sieves, carries with 3 heat of 60 liters of branches of 70% ethanol, extracts 2h first, and 1h is respectively extracted in second and third time, merges No. 3 times extracting solution, is concentrated into about 8 liters.Concentrated solution is diluted with water to 25 liters, adds 5 liters of 1% chitosan solutions under the stirring at low speed, leaves standstill 1h, filters, and filtrate is adjusted to pH=8;
(2) above-mentioned filtrate by 10 liters of HP20 type macroporous resin column, is removed impurity with 5BV water and 4BV 30% ethanol drip washing with the flow velocity of 2BV/h (BV is the column volume of macroporous resin) successively, uses 4BV 85% ethanol elution again, collects elutriant, concentrates near doing.Concentrated solution by 2 liters of HP20 type macroporous resin column, repeats above-mentioned elution program with 20% alcohol dilution to 2 liter, and concentrate eluant gets macroporous resin extract to doing;
(3) with macroporous resin extract with etc. the octadecyl silane (ODS) of weight mix, last ODS post with 25% ethanol drip washing impurity, is used 65% ethanol elution again, be in charge of collection, know respectively to manage with thin-layer chromatography inspection and have or not Isoforskolin in the elutriant, merge the elutriant that contains Isoforskolin, be concentrated into dried, repeat above ODS column chromatography, collection contains the elutriant of Isoforskolin, and elutriant is concentrated into dried, gets the ODS extract;
(4) ODS extract acetic acid ethyl dissolution after waiting to separate out white crystal, leaches crystallization, and crystallization is carried out recrystallization after with acetic acid ethyl dissolution, leaches crystal, and drying gets purity greater than 90% Isoforskolin.
(5) preparation liquid phase purifying: above-mentioned Isoforskolin crystal with dissolve with methanol after, adopt the preparation liquid phase to be further purified liquid phase chromatogram condition: chromatographic column is the ODS post, and moving phase is water methanol (35: 65), detects wavelength 210nm.Collect the effluent liquid of main peak retention time, be concentrated into driedly, get purity greater than 98% Isoforskolin.
Embodiment 2
(1) the Coleus forskohlii Briq. herb is 5 kilograms, pulverizes, and crosses 20 mesh sieves, with diacolation behind the 80% alcohol immersion 12h, collects 40 liters of percolates, is concentrated into about 5 liters.Concentrated solution is diluted with water to 20 liters, filters, and filtrate is adjusted to pH=9;
(2) above-mentioned filtrate by 10 liters of HZ818 type macroporous resin column, is removed impurity with 5BV water and 4BV 25% ethanol drip washing with the flow velocity of 2BV/h (BV is the column volume of macroporous resin) successively, uses 4BV 80% ethanol elution again, collects elutriant, concentrates near doing.Concentrated solution by 2 liters of HZ818 type macroporous resin column, repeats above-mentioned elution program with 20% alcohol dilution to 2 liter, and concentrate eluant gets macroporous resin extract to doing;
(3) with macroporous resin extract with after the small amount of ethanol dissolving, last ODS post with 20% ethanol drip washing impurity, is used 60% ethanol elution again, be in charge of collection, know respectively to manage with thin-layer chromatography inspection and have or not Isoforskolin in the elutriant, merge the elutriant that contains Isoforskolin, be concentrated into dried, repeat above ODS column chromatography, collection contains the elutriant of Isoforskolin, and elutriant is concentrated into dried, gets the ODS extract;
(4) ODS extract anhydrous alcohol solution after waiting to separate out white crystal, leaches crystallization.Crystallization is carried out recrystallization after with anhydrous alcohol solution, leaches crystal, and drying gets purity greater than 90% Isoforskolin;
(5) preparation liquid phase purifying: above-mentioned Isoforskolin crystal with dissolve with methanol after, adopt the preparation liquid phase to be further purified liquid phase chromatogram condition: chromatographic column is the ODS post, and moving phase is water methanol (35: 65), detects wavelength 210nm.Collect the effluent liquid of main peak retention time, be concentrated into driedly, get purity greater than 98% Isoforskolin.

Claims (6)

1.一种提取毛喉鞘蕊花有效成分的方法，该有效成分为异佛司可林，其特征在于，其包括步骤：1. a method for extracting the active ingredient of Coleus forskohlii, the active ingredient is Isoferskolin, it is characterized in that, it comprises steps: (1)提取：取毛喉鞘蕊花全草或根粉碎后，用70～95％乙醇提取，浓缩提取液，浓缩液直接过滤，或加入絮凝剂后过滤，滤液备用；(1) Extraction: take the whole herb or root of Coleus forskohlii and pulverize it, extract with 70-95% ethanol, concentrate the extract, filter the concentrate directly, or add a flocculant to filter, and use the filtrate for later use; (2)大孔树脂柱层析纯化：上述滤液调节pH值后通过大孔树脂柱，依次用水及浓度为20～40％乙醇淋洗，再用60～85％乙醇洗脱，将洗脱液浓缩至干，得大孔树脂提取物；(2) Purification by macroporous resin column chromatography: the above-mentioned filtrate passes through the macroporous resin column after adjusting the pH value, rinses with water and 20-40% ethanol successively, and then elutes with 60-85% ethanol. Concentrate to dryness to obtain macroporous resin extract; (3)十八烷基键合硅胶(ODS)柱层析纯化：取上述大孔树脂提取物，与等重量ODS混合均匀，或用尽可能少量的乙醇溶解后，上ODS柱纯化，以20～35％乙醇淋洗杂质，再用40～65％乙醇洗脱，将洗脱液浓缩至干，得ODS提取物；(3) Octadecyl bonded silica gel (ODS) column chromatography purification: take the above-mentioned macroporous resin extract, mix with equal weight ODS, or after dissolving with as little ethanol as possible, go to ODS column purification, with 20 Rinse impurities with ~35% ethanol, then elute with 40~65% ethanol, and concentrate the eluent to dryness to obtain ODS extract; (4)重结晶：上述ODS提取物以乙酸乙酯、丙酮、乙醇或所述溶剂的混合液进行重结晶，析出白色晶体后，滤出结晶，干燥，得纯度大于90％的异佛司可林；(4) Recrystallization: the above-mentioned ODS extract is recrystallized with ethyl acetate, acetone, ethanol or the mixed solution of the solvent, after the white crystals are separated out, the crystals are filtered out and dried to obtain Isofosco with a purity greater than 90%. Forest; (5)制备液相纯化：制得的异佛司可林晶体用甲醇溶解后，经制备液相进一步纯化，得纯度大于98％的异佛司可林。(5) Purification in the preparative liquid phase: after dissolving the prepared isofaskolin crystals with methanol, further purify in the preparative liquid phase to obtain isofaskolin with a purity greater than 98%. 2.根据权利要求1所述的方法，其特征在于，所述步骤(2)中的大孔树脂选自非极性或弱极性的聚苯乙烯型多孔性吸附树脂。2. The method according to claim 1, characterized in that the macroporous resin in the step (2) is selected from nonpolar or weakly polar polystyrene type porous adsorption resins. 3.根据权利要求1所述的方法，其特征在于，所述步骤(2)中的上大孔树脂前的滤液pH值范围为5～10。3. The method according to claim 1, characterized in that the pH range of the filtrate before applying the macroporous resin in the step (2) is 5-10. 4.根据权利要求1所述的方法，其特征在于，所述步骤(2)中大孔树脂柱层析纯化进行1～3次。4. The method according to claim 1, characterized in that the macroporous resin column chromatography purification in the step (2) is carried out 1 to 3 times. 5.根据权利要求1所述的方法，其特征在于，所述步骤(3)中的ODS柱层析纯化进行1～3次。5. The method according to claim 1, characterized in that the ODS column chromatography purification in the step (3) is performed 1 to 3 times. 6.根据权利要求1所述的方法，其特征在于，所述步骤(4)中的重结晶进行2～3次。6. The method according to claim 1, characterized in that, the recrystallization in the step (4) is carried out 2 to 3 times.
CN2011104588184A 2011-12-29 2011-12-29 Method for extracting effective component of coleus forskohlii Pending CN103183658A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103588745A (en) * 2013-11-26 2014-02-19 陕西嘉禾植物化工有限责任公司 Method for extracting and separating forskolin in coleus forskohlii
CN105693684A (en) * 2016-01-22 2016-06-22 昆明医科大学 Isoforskolin preparing method
CN106596797A (en) * 2017-01-19 2017-04-26 湖北福人药业股份有限公司 Content determination of coleus forskohlii capsules
CN115536665A (en) * 2022-09-14 2022-12-30 河北省药品医疗器械检验研究院(河北省化妆品检验研究中心) Separation method of high-purity cepharanthine

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CN1651423A (en) * 2004-11-26 2005-08-10 复旦大学 Coleus forskohlii extract and its preparation method and use
CN101259123A (en) * 2008-04-18 2008-09-10 昆明医学院 Application of isoforskolin and its analogue in preventing and controlling lung injury

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CN1651423A (en) * 2004-11-26 2005-08-10 复旦大学 Coleus forskohlii extract and its preparation method and use
CN101259123A (en) * 2008-04-18 2008-09-10 昆明医学院 Application of isoforskolin and its analogue in preventing and controlling lung injury

Non-Patent Citations (1)

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Title
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103588745A (en) * 2013-11-26 2014-02-19 陕西嘉禾植物化工有限责任公司 Method for extracting and separating forskolin in coleus forskohlii
CN103588745B (en) * 2013-11-26 2015-10-28 陕西嘉禾植物化工有限责任公司 A kind of method of Extraction and separation not this Kelin from Coleus forskohlii Briq.
CN105693684A (en) * 2016-01-22 2016-06-22 昆明医科大学 Isoforskolin preparing method
CN106596797A (en) * 2017-01-19 2017-04-26 湖北福人药业股份有限公司 Content determination of coleus forskohlii capsules
CN106596797B (en) * 2017-01-19 2020-04-21 湖北福人药业股份有限公司 Content determination of Coleus forskohlii capsule
CN115536665A (en) * 2022-09-14 2022-12-30 河北省药品医疗器械检验研究院(河北省化妆品检验研究中心) Separation method of high-purity cepharanthine
CN115536665B (en) * 2022-09-14 2023-12-01 河北省药品医疗器械检验研究院(河北省化妆品检验研究中心) Separation method of high-purity stephanine
WO2024055459A1 (en) * 2022-09-14 2024-03-21 河北省药品医疗器械检验研究院(河北省化妆品检验研究中心) Method for separating high-purity cepharanthine

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Application publication date: 20130703