CN104961745A - Preparation method of isobrucein B - Google Patents

Preparation method of isobrucein B Download PDF

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Publication number
CN104961745A
CN104961745A CN201510270792.9A CN201510270792A CN104961745A CN 104961745 A CN104961745 A CN 104961745A CN 201510270792 A CN201510270792 A CN 201510270792A CN 104961745 A CN104961745 A CN 104961745A
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China
Prior art keywords
bruceine
different
preparation
phase
extract
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CN201510270792.9A
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刘东锋
杨成东
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Nanjing Zelang Medical Technology Co Ltd
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Nanjing Zelang Medical Technology Co Ltd
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Priority to CN201510270792.9A priority Critical patent/CN104961745A/en
Publication of CN104961745A publication Critical patent/CN104961745A/en
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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07DHETEROCYCLIC COMPOUNDS
    • C07D493/00Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
    • C07D493/02Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
    • C07D493/10Spiro-condensed systems
    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00Technologies relating to chemical industry
    • Y02P20/50Improvements relating to the production of bulk chemicals
    • Y02P20/54Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

The invention relates to a simple-operation and less-pollution preparation method of isobrucein B. The method comprises the following steps: 1, crushing branches and leaves of Brucea antidysenterica as a raw material, extracting by adopting supercritical CO2, and collecting the obtained extract; 2, carrying out chloroform-methanol gradient elution with silica gel as a packing, collecting the obtained target eluate, concentrating, and drying to obtain a crude extract product; and 3, separating and purifying isobrucein B in the crude extract product by adopting high-speed countercurrent chromatography with the two-phase solvent system of chloroform-methanol-water, the upper phase of a stationary phase and the lower phase of a mobile phase, collecting an isobrucein B component, carrying out vacuum concentration, and drying to obtain isobrucein B. The isobrucein B prepared in the invention has high purity and good quality.

Description

A kind of preparation method of different bruceine B
Technical field
The invention belongs to traditional Chinese medicine extraction separation technology field, relate to a kind of method being separated the different bruceine B of preparation from the branches and leaves of anti-dysentery kosam seeds.
Background technology
Anti-dysentery kosam seeds (Brucea antidysenterica Mill) is the one tree of Simarubaceae brucea, is used for for cancer in Ethiopia.Different bruceine B is the principal character chemical composition of these trees, is needle crystal, is dissolved in the organic solvent such as ether, methylene dichloride, and fusing point is 243 ~ 246 DEG C.
Modern study shows, different bruceine B has multiple biological activity, as desinsection stops dysentery, antimalarial, the effect such as antitumor, antiviral.Very strong inhibit activities is had, LC to the multiple cancer cells of human body 50at 100 ~ 10 μMs.
By document chains, the domestic technique preparing different bruceine B from anti-dysentery kosam seeds that there is not yet is reported.
Summary of the invention
The object of this invention is to provide a kind of preparation method of different bruceine B.
The object of the invention is to be achieved through the following technical solutions:
(1) with the branches and leaves of anti-dysentery kosam seeds for raw material, through pulverization process, adopt supercritical CO 2extract, with the ethanol of total solvent volume 4% for entrainment agent, arranging extraction temperature is 40 ~ 50 DEG C, after temperature-stable, pass into CO 2, flow is 25 ~ 35L/h, and regulates pressure to 30 ~ 40MPa, after carrying out extraction 2 ~ 3h, collects extract;
(2) the concentrated rear upper silicagel column of extract, take volume ratio as the chloroform-methanol gradient elution of 50:3, collect target elutriant, concentrate drying obtains crude extract;
(3) the different bruceine B in high-speed countercurrent chromatography separation and purification crude extract is adopted, its two-phase solvent system is chloroform-methanol-water, upper is stationary phase mutually, rotate main frame, pump into and do moving phase mutually down, moving phase dissolves crude extract by sampling valve sample introduction, UV-detector on-line monitoring, collect different bruceine B component, vacuum concentration, namely cryodrying obtains different bruceine B.
Silica gel order number described in step (2) is 200 ~ 300 orders.
The volume ratio of the two-phase solvent described in step (3) is 15:4:1.
High-speed counter-current chromatograph rotating speed described in step (3) is 1000rpm, and flow rate of mobile phase is 4 ~ 5mL/min.
Ultraviolet detection wavelength described in step (3) is 242nm.
Low temperature described in step (3) is 0 ~ 5 DEG C.
Beneficial effect of the present invention is: adopt supercritical extraction, extraction time is short, extraction efficiency is high, nontoxic, tasteless, cheap and easy to get; Adopt silica gel column chromatography enriching and purifying, can plurality of impurities be removed, alleviate the workload of operation below; Adopt high speed adverse current chromatogram good separating effect, product purity is high.
Further illustrate the present invention below in conjunction with embodiment, but the scope of protection of present invention is not limited to following embodiments.
Embodiment
Embodiment 1:
With the branches and leaves of anti-dysentery kosam seeds for raw material, through pulverization process, adopt supercritical CO 2extract, with the ethanol of total solvent volume 4% for entrainment agent, arranging extraction temperature is 40 DEG C, after temperature-stable, pass into CO 2, flow is 25L/h, and regulates pressure to 30MPa, after carrying out extraction 2h, collects extract; After extract is concentrated, upper order number is the silicagel column of 200 ~ 300, take volume ratio as the chloroform-methanol gradient elution of 50:3, and collect target elutriant, concentrate drying obtains crude extract; Different bruceine B in employing high-speed countercurrent chromatography separation and purification crude extract take volume ratio as the chloroform-methanol-water of 15:4:1 is two-phase solvent system, and upper is stationary phase mutually, rotate main frame, pump into and do moving phase mutually down, flow rate of mobile phase is 4mL/min, and moving phase dissolves crude extract by sampling valve sample introduction, UV-detector on-line monitoring, determined wavelength is 242nm, collects different bruceine B component, vacuum concentration, at 0 DEG C, be drying to obtain different bruceine B, content is 98.4%.
Embodiment 2:
With the branches and leaves of anti-dysentery kosam seeds for raw material, through pulverization process, adopt supercritical CO 2extract, with the ethanol of total solvent volume 4% for entrainment agent, arranging extraction temperature is 45 DEG C, after temperature-stable, pass into CO 2, flow is 30L/h, and regulates pressure to 35MPa, after carrying out extraction 3h, collects extract; After extract is concentrated, upper order number is the silicagel column of 200 ~ 300, take volume ratio as the chloroform-methanol gradient elution of 50:3, and collect target elutriant, concentrate drying obtains crude extract; Different bruceine B in employing high-speed countercurrent chromatography separation and purification crude extract take volume ratio as the chloroform-methanol-water of 15:4:1 is two-phase solvent system, and upper is stationary phase mutually, rotate main frame, pump into and do moving phase mutually down, flow rate of mobile phase is 4.5mL/min, and moving phase dissolves crude extract by sampling valve sample introduction, UV-detector on-line monitoring, determined wavelength is 242nm, collects different bruceine B component, vacuum concentration, at 5 DEG C, be drying to obtain different bruceine B, content is 99.0%.
Embodiment 3:
With the branches and leaves of anti-dysentery kosam seeds for raw material, through pulverization process, adopt supercritical CO 2extract, with the ethanol of total solvent volume 4% for entrainment agent, arranging extraction temperature is 50 DEG C, after temperature-stable, pass into CO 2, flow is 35L/h, and regulates pressure to 40MPa, after carrying out extraction 3h, collects extract; After extract is concentrated, upper order number is the silicagel column of 200 ~ 300, take volume ratio as the chloroform-methanol gradient elution of 50:3, and collect target elutriant, concentrate drying obtains crude extract; Different bruceine B in employing high-speed countercurrent chromatography separation and purification crude extract take volume ratio as the chloroform-methanol-water of 15:4:1 is two-phase solvent system, and upper is stationary phase mutually, rotate main frame, pump into and do moving phase mutually down, flow rate of mobile phase is 5mL/min, and moving phase dissolves crude extract by sampling valve sample introduction, UV-detector on-line monitoring, determined wavelength is 242nm, collects different bruceine B component, vacuum concentration, at 5 DEG C, be drying to obtain different bruceine B, content is 98.1%.

Claims (6)

1. a preparation method for different bruceine B, is characterized in that, comprises the following steps:
(1) with the branches and leaves of anti-dysentery kosam seeds for raw material, through pulverization process, adopt supercritical CO 2extract, with the ethanol of total solvent volume 4% for entrainment agent, arranging extraction temperature is 40 ~ 50 DEG C, after temperature-stable, pass into CO 2, flow is 25 ~ 35L/h, and regulates pressure to 30 ~ 40MPa, after carrying out extraction 2 ~ 3h, collects extract;
(2) the concentrated rear upper silicagel column of extract, take volume ratio as the chloroform-methanol gradient elution of 50:3, collect target elutriant, concentrate drying obtains crude extract;
(3) the different bruceine B in high-speed countercurrent chromatography separation and purification crude extract is adopted, its two-phase solvent system is chloroform-methanol-water, upper is stationary phase mutually, rotate main frame, pump into and do moving phase mutually down, moving phase dissolves crude extract by sampling valve sample introduction, UV-detector on-line monitoring, collect different bruceine B component, vacuum concentration, namely cryodrying obtains different bruceine B.
2. the preparation method of a kind of different bruceine B according to claim 1, is characterized in that, the silica gel order number described in step (2) is 200 ~ 300 orders.
3. the preparation method of a kind of different bruceine B according to claim 1, is characterized in that, the volume ratio of the two-phase solvent described in step (3) is 15:4:1.
4. the preparation method of a kind of different bruceine B according to claim 1, is characterized in that, the high-speed counter-current chromatograph rotating speed described in step (3) is 1000rpm, and flow rate of mobile phase is 4 ~ 5mL/min.
5. the preparation method of a kind of different bruceine B according to claim 1, is characterized in that, the ultraviolet detection wavelength described in step (3) is 242nm.
6. the preparation method of a kind of different bruceine B according to claim 1, is characterized in that, the low temperature described in step (3) is 0 ~ 5 DEG C.
CN201510270792.9A 2015-05-26 2015-05-26 Preparation method of isobrucein B Pending CN104961745A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510270792.9A CN104961745A (en) 2015-05-26 2015-05-26 Preparation method of isobrucein B

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Application Number Priority Date Filing Date Title
CN201510270792.9A CN104961745A (en) 2015-05-26 2015-05-26 Preparation method of isobrucein B

Publications (1)

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CN104961745A true CN104961745A (en) 2015-10-07

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113024551A (en) * 2021-05-20 2021-06-25 江西中医药大学 Novel compound extracted and separated from brucea javanica, and preparation method and application thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113024551A (en) * 2021-05-20 2021-06-25 江西中医药大学 Novel compound extracted and separated from brucea javanica, and preparation method and application thereof

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Application publication date: 20151007