CN102863456A - Method for preparing Wikstroelide D - Google Patents

Method for preparing Wikstroelide D Download PDF

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Publication number
CN102863456A
CN102863456A CN2012103666617A CN201210366661A CN102863456A CN 102863456 A CN102863456 A CN 102863456A CN 2012103666617 A CN2012103666617 A CN 2012103666617A CN 201210366661 A CN201210366661 A CN 201210366661A CN 102863456 A CN102863456 A CN 102863456A
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CN
China
Prior art keywords
extraction
ester
canescent wikstroemia
preparing
wikstroemia
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CN2012103666617A
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Chinese (zh)
Inventor
苏刘花
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Nanjing Zelang Agricultural Development Co Ltd
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Nanjing Zelang Agricultural Development Co Ltd
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Priority to CN2012103666617A priority Critical patent/CN102863456A/en
Publication of CN102863456A publication Critical patent/CN102863456A/en
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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
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • 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
    • 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
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • 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
    • Y02P20/582—Recycling of unreacted starting or intermediate materials

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  • Medicines Containing Plant Substances (AREA)
  • Extraction Or Liquid Replacement (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

The invention belongs to the field of natural drug chemistry, and relates to a method for preparing Wikstroelide D. The method comprises the following steps: pulverizing Wikstroemia retusa A.Gray bark while controlling the particle size at 40-80 meshes, adding into an extraction kettle, carrying out CO2 extraction by using a 90-100 vol% ethanol water solution as an entrainer, collecting the extract, recovering the ethanol, degreasing with heptane, carrying out high-speed counter-current chromatography separation by using n-hexane-ethyl acetate-methanol-water as a solvent system, and drying the fraction under reduced pressure to obtain the Wikstroelide D. The Wikstroelide D product prepared by the method provided by the invention has the advantages of no pollution and high yield; and the method is suitable for industrialized preparation of Wikstroelide D.

Description

A kind of method for preparing canescent wikstroemia ester D
Technical field
The invention belongs to the Natural Medicine Chemistry field, particularly relate to a kind of employing supercritical CO 2Extraction and high speed adverse current chromatogram isolation technique prepare the method for canescent wikstroemia ester D.
Background technology
Canescent wikstroemia ester D(Wikstroelide D) be diterpene-kind compound, molecular formula C 52H 80O 11, molecular weight 881.2.Canescent wikstroemia ester D extensively exists in natural phant, mainly from Thymelaeceae plant nick canescent wikstroemia Wikstroemia retusaSeparate in the A.Gray bark and obtain.Pharmacological research shows, canescent wikstroemia ester D has cytotoxic activity, in the MTT test, P388 and people's lung cancer strain PC-6 is cytotoxic activity.
At present, domesticly there is not yet a large amount of methods reports that prepare canescent wikstroemia ester D.
Summary of the invention
The object of the present invention is to provide a kind of method for preparing canescent wikstroemia ester D.
To achieve these goals, technical scheme of the present invention is as follows:
A kind of method for preparing canescent wikstroemia ester D is characterized in that may further comprise the steps: get nick canescent wikstroemia bark and pulverize, the control particle diameter is the 40-80 order, adds in the extraction kettle, and the aqueous ethanolic solution take volume percent as 90-100% is entrainment agent, adopts supercritical CO 2Extraction liquid is collected in extraction, uses the heptane degreasing behind the Recycled ethanol, and again take normal hexane-ethyl acetate-methanol-water as solvent systems, upper is stationary phase mutually, and lower is moving phase mutually, adopts high speed adverse current chromatogram to separate, and the flow point drying under reduced pressure namely gets canescent wikstroemia ester D.
Described powder particle diameter is 40 orders, and the aqueous ethanolic solution volume percent is 95%.
Described supercritical CO 2Extraction conditions is: extraction temperature is 42-55 ℃, and extracting pressure is 25-30MPa, and separating still I temperature is 35-45 ℃, pressure is 8-10MPa, and separating still II temperature is 28-38 ℃, and pressure is 6-7MPa, the entrainment agent consumption is that the 1-2 of raw material doubly measures, and extraction time is 2-4 hour, CO 2Flow is 18-22L/h.
Described normal hexane-ethyl acetate-methanol-water solvent systems ratio is (1-3): (4-6): (2-4): (2-4), the chromatographic instrument rotating speed is 800-900rpm.
Advantage of the present invention is: the process using supercritical CO 2Extraction separates canescent wikstroemia ester D with high speed adverse current chromatogram, the loss of sepn process product-free, and preparation cycle is short, reagent can recycling, preparation amount is large, and is pollution-free, overcome that sample extremely adsorbs in the conventional post chromatographic separation process, the technological deficiencies such as technique is loaded down with trivial details, poor reproducibility.
Embodiment:
Further specify the present invention below in conjunction with embodiment, but the scope of protection of present invention is not limited to following embodiment.
Embodiment 1:
Get nick canescent wikstroemia bark and pulverized 40 orders, get the 1kg raw material and add supercritical CO 2In the extraction kettle, opening device, adjust device parameter: extraction temperature is 55 ℃, and extracting pressure is 30MPa, and separating still I temperature is 45 ℃, and pressure is 9MPa, and separating still II temperature is 35 ℃, and pressure is 7MPa, passes into CO 2Fluid, reach above-mentioned parameter after, with CO 2Liquid 20L/h flow velocity extraction, and add and carry 95% aqueous ethanolic solution 1.2L secretly, dynamic extraction 4h collects extraction liquid, and Recycled ethanol adds the heptane degreasing, obtains crude extract.Get normal hexane, ethyl acetate, methyl alcohol, water, mix for 1:5:3:3 in proportion, get and pump into mutually separator column, until being fixed, fills mutually by separator column, rotating speed is 850rpm, simultaneously lower flow velocity with 6ml/min is pumped into, flow out from outlet end and after two-phase set up running balance, flow rate of mobile phase was adjusted to 2.0ml/min until moving phase, inject the crude extract of moving phase dissolving, the UV-detector monitoring, the monitoring wavelength is 231nm, collects canescent wikstroemia ester D flow point, continuous separate from, merge the flow point drying under reduced pressure and get canescent wikstroemia ester D, high performance liquid phase detects, content 94.0%.
Embodiment 2:
Get nick canescent wikstroemia bark and pulverized 40 orders, get the 1kg raw material and add supercritical CO 2In the extraction kettle, opening device, adjust device parameter: extraction temperature is 42 ℃, and extracting pressure is 28MPa, and separating still I temperature is 40 ℃, and pressure is 10MPa, and separating still II temperature is 32 ℃, and pressure is 6.5MPa, passes into CO 2Fluid, reach above-mentioned parameter after, with CO 2The extraction of liquid 18L/h flow velocity, and carry 95% aqueous ethanolic solution 1.8L secretly, dynamic extraction 3h collects extraction liquid, and Recycled ethanol adds the heptane degreasing, obtains crude extract.Get normal hexane, ethyl acetate, methyl alcohol, water, mix for 3:6:4:4 in proportion, get and pump into mutually separator column, until being fixed, fills mutually by separator column, rotating speed is 800rpm, simultaneously lower flow velocity with 10ml/min is pumped into, flow out from outlet end and after two-phase set up running balance, flow rate of mobile phase was adjusted to 3.0ml/min until moving phase, inject the crude extract of moving phase dissolving, the UV-detector monitoring, the monitoring wavelength is 231nm, collects canescent wikstroemia ester D flow point, continuous separate from, merge the flow point drying under reduced pressure and get canescent wikstroemia ester D, high performance liquid phase detects, content 91.5%.
Embodiment 3:
Get nick canescent wikstroemia bark and pulverized 40 orders, get the 1kg raw material and add supercritical CO 2In the extraction kettle, opening device, adjust device parameter: extraction temperature is 45 ℃, and extracting pressure is 25MPa, and separating still I temperature is 35 ℃, and pressure is 8MPa, and separating still II temperature is 28 ℃, and pressure is 6MPa, passes into CO 2Fluid, reach above-mentioned parameter after, with CO 2The extraction of liquid 19L/h flow velocity, and carry 95% aqueous ethanolic solution 2L secretly, dynamic extraction 2h collects extraction liquid, and Recycled ethanol adds the heptane degreasing, obtains crude extract.Get normal hexane, ethyl acetate, methyl alcohol, water, mix for 2:4:2:3 in proportion, get and pump into mutually separator column, until being fixed, fills mutually by separator column, rotating speed is 900rpm, simultaneously lower flow velocity with 5ml/min is pumped into, flow out from outlet end and after two-phase set up running balance, flow rate of mobile phase was adjusted to 1.5ml/min until moving phase, inject the crude extract of moving phase dissolving, the UV-detector monitoring, the monitoring wavelength is 231nm, collects canescent wikstroemia ester D flow point, continuous separate from, merge the flow point drying under reduced pressure and get canescent wikstroemia ester D product, high performance liquid phase detects, content 93.4%.
Embodiment 4:
Get nick canescent wikstroemia bark and pulverized 40 orders, get the 1kg raw material and add supercritical CO 2In the extraction kettle, opening device, adjust device parameter: extraction temperature is 55 ℃, and extracting pressure is 30MPa, and separating still I temperature is 45 ℃, and pressure is 9MPa, and separating still II temperature is 38 ℃, and pressure is 7MPa, passes into CO 2Fluid, reach above-mentioned parameter after, with CO 2The extraction of liquid 20L/h flow velocity, and carry 95% aqueous ethanolic solution 1L secretly, dynamic extraction 4h collects extraction liquid, and Recycled ethanol adds the heptane degreasing, obtains crude extract.Get normal hexane, ethyl acetate, methyl alcohol, water, mix for 2:5:3:2 in proportion, get and pump into mutually separator column, until being fixed, fills mutually by separator column, rotating speed is 850rpm, simultaneously lower flow velocity with 8ml/min is pumped into, flow out from outlet end and after two-phase set up running balance, flow rate of mobile phase was adjusted to 2.0ml/min until moving phase, inject the crude extract of moving phase dissolving, the UV-detector monitoring, the monitoring wavelength is 231nm, collects canescent wikstroemia ester D flow point, continuous separate from, merge the flow point drying under reduced pressure and get canescent wikstroemia ester D, high performance liquid phase detects, content 94.3%.
Embodiment 5:
Get nick canescent wikstroemia bark and pulverized 40 orders, get the 1kg raw material and add supercritical CO 2In the extraction kettle, opening device, adjust device parameter: extraction temperature is 50 ℃, and extracting pressure is 30MPa, and separating still I temperature is 45 ℃, and pressure is 9MPa, and separating still II temperature is 37 ℃, and pressure is 7MPa, passes into CO 2Fluid, reach above-mentioned parameter after, with CO 2The extraction of liquid 22L/h flow velocity, and carry 75% aqueous ethanolic solution 1.5L secretly, dynamic extraction 4h collects extraction liquid, and Recycled ethanol adds the heptane degreasing, obtains crude extract.Get normal hexane, ethyl acetate, methyl alcohol, water, mix for 3:5:4:4 in proportion, get and pump into mutually separator column, until being fixed, fills mutually by separator column, rotating speed is 850rpm, simultaneously lower flow velocity with 10ml/min is pumped into, flow out from outlet end and after two-phase set up running balance, flow rate of mobile phase was adjusted to 2.5ml/min until moving phase, inject the crude extract of moving phase dissolving, the UV-detector monitoring, the monitoring wavelength is 231nm, collects canescent wikstroemia ester D flow point, continuous separate from, merge the flow point drying under reduced pressure and get canescent wikstroemia ester D, high performance liquid phase detects, content 90.2%.

Claims (4)

1. method for preparing canescent wikstroemia ester D is characterized in that may further comprise the steps: get nick canescent wikstroemia bark and pulverize, the control particle diameter is the 40-80 order, adds in the extraction kettle, and the aqueous ethanolic solution take volume percent as 90-100% is entrainment agent, adopts supercritical CO 2Extraction liquid is collected in extraction, uses the heptane degreasing behind the Recycled ethanol, and again take normal hexane-ethyl acetate-methanol-water as solvent systems, upper is stationary phase mutually, and lower is moving phase mutually, adopts high speed adverse current chromatogram to separate, and the flow point drying under reduced pressure namely gets canescent wikstroemia ester D.
2. the described method for preparing canescent wikstroemia ester D according to claim 1 is characterized in that described powder particle diameter is 40 orders, and the aqueous ethanolic solution volume percent is 95%.
3. the described method for preparing canescent wikstroemia ester D according to claim 1 is characterized in that described supercritical CO 2Extraction conditions is: extraction temperature is 42-55 ℃, and extracting pressure is 25-30MPa, and separating still I temperature is 35-45 ℃, pressure is 8-10MPa, and separating still II temperature is 28-38 ℃, and pressure is 6-7MPa, the entrainment agent consumption is that the 1-2 of raw material doubly measures, and extraction time is 2-4 hour, CO 2Flow is 18-22L/h.
4. the described method for preparing canescent wikstroemia ester D according to claim 1, it is characterized in that described normal hexane-ethyl acetate-methanol-water solvent systems ratio is (1-3): (4-6): (2-4): (2-4), the chromatographic instrument rotating speed is 800-900rpm.
CN2012103666617A 2012-09-28 2012-09-28 Method for preparing Wikstroelide D Pending CN102863456A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102311444A (en) * 2011-07-05 2012-01-11 南京泽朗农业发展有限公司 Method for extracting Sanggenon D from Cortex Mori
EP2468741A1 (en) * 2010-12-16 2012-06-27 Bel/Novamann International s.r.o. Novel quercetin derivatives, their preparation, pharmaceutical compositions containing them and their use

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2468741A1 (en) * 2010-12-16 2012-06-27 Bel/Novamann International s.r.o. Novel quercetin derivatives, their preparation, pharmaceutical compositions containing them and their use
CN102311444A (en) * 2011-07-05 2012-01-11 南京泽朗农业发展有限公司 Method for extracting Sanggenon D from Cortex Mori

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
FUMIKO ABE，等: "DAPHNANE DITERPENOIDS FROM THE BARK OF WIKSTROEMIA RETUSA", 《PHYTOCHEMISTRY》 *
FUMIKO ABE，等: "MINOR DAPHNANE-TYPE DITERPENOIDS FROM WIKSTROEMIA RETUSA", 《PHYTOCHEMISTRY》 *

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Application publication date: 20130109