CN102146083B - Method for separating and extracting cepharanthine - Google Patents

Method for separating and extracting cepharanthine Download PDF

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Publication number
CN102146083B
CN102146083B CN 201110056911 CN201110056911A CN102146083B CN 102146083 B CN102146083 B CN 102146083B CN 201110056911 CN201110056911 CN 201110056911 CN 201110056911 A CN201110056911 A CN 201110056911A CN 102146083 B CN102146083 B CN 102146083B
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cepharanthine
solution
aqueous solution
elutriant
hours
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CN102146083A (en
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蔡群虎
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HANDE BIOLOGICAL TECHNOLOGY Co Ltd YUNNAN
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HANDE BIOLOGICAL TECHNOLOGY Co Ltd YUNNAN
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Abstract

The invention relates to a method for separating and extracting cepharanthine, which comprises the following steps of: taking a dry stephania tuberous root, crushing, adding 2 to 3L of water and 0.5 to 5ml of mixed bio-enzyme liquid into each kilogram of crushed material, soaking for 1 hour, adding 9 to 11L of 1 to 2 mass percent aqueous solution of HCL, heating to 90DEG C, and soaking for 3 hours; and separating out a liquid medicine, adding 9 to 11L of 1 to 2 percent aqueous solution of HCL into dregs, decocting for 3 hours, filtering, mixing filtrate, adsorbing by adopting a cation exchange column or cation cellulose column, enriching the cepharanthine, eluting by using solution that sodium hydroxide is added into 60 to 90 percent aqueous solution of ethanol until the saturation state as eluent, collecting eluent of a cepharanthine section, regulating the pH value to be 7, concentrating for recovering ethanol, filtering, dissolving a precipitate by using absolute ethanol, eluting and purifying by using an anion column, collecting eluent of a cepharanthine section, concentrating, crystallizing, and drying to obtain the cepharanthine. The method has the advantages of simple operation, light pollution, low cost, high yield, high purity of cepharanthine, and suitability for industrial production.

Description

A kind of separating and extracting method of cepharanthine
Technical field
The present invention relates to a kind of extraction and separation method of cepharanthine, specifically, is a kind of method of extracting cepharanthine from the plant Root of Epigeal Srephaia.
Background technology
Cepharanthine (Cepharanthine) is a kind of bioactive imidazole alkaloid that has, and is mainly derived from the menispermaceae plant, Root of Epigeal Srephaia for example, root of Smoothleaf Stephania, Radix Stephaniae Cepharanthae, Radix Stephaniae excentricae and root of Taiwan Stephania etc.Because the impact of the factors such as geographical environment, weather condition, the same quality of medicinal material of different sources there are differences more, and the difference in kind and the place of production, source causes that cepharanthine content just differs in the plant.
Cepharanthine can promote myelosis, increases the quantity of blood middle leukocytes, is applicable to leukopenia; Simultaneously also be anticarcinogen synergistic agent preferably, can improve the curative effect of cancer therapy drug, reduce the consumption of anticarcinogen, alleviate toxic side effect; Many animals inflammatory edema and pain all there are obvious restraining effect, and without any toxic reaction.
Cepharanthine is mainly slightly carried by pure methyl alcohol or ethanol in the prior art, the molten alkali of acid is heavy, silica gel column chromatography or alumina column chromatography, the acetone crystallization, the separating obtained cepharanthine rate of recovery of these traditional method for extracting is lower, in the purge process cepharanthine loss larger, the organic solvent consumption is large, the industrialization cost is higher, and industrial pollution is serious.
Summary of the invention
The invention provides a kind of cepharanthine preparation method who is applicable to suitability for industrialized production.
Technical scheme of the present invention is:
Get and do not allow dryly the piece root, pulverize, the per kilogram crushed material added 2-3L water and 0.5-5ml mixed biologic enzyme vacuole after 1 hour, added the 1-2%(mass percent of 9-11 L, lower with) the HCL aqueous solution is heated to 90 ℃ and soaked 3 hours; Separate liquid, the 1-2%HCL aqueous solution that the dregs of a decoction add 9-11 L again decocted 3 hours, filtered.Merge 2 times filtrate, adopt the absorption of cationic exchange coloum or cationic cellulose post, the enrichment cepharanthine, with adding sodium hydroxide to the 60-90%(mass percent, lower together) in the aqueous ethanolic solution to the solution of state of saturation as elutriant, wash-out, collect cepharanthine section elutriant, regulate pH value to 7, concentration and recovery ethanol, filter, throw out with anhydrous alcohol solution after, with anion column wash-out purifying, collect cepharanthine section elutriant, concentrated, crystallization is drying to obtain cepharanthine.
Mixed biologic enzyme liquid is by hemicellulase, pectin lyase, middle temperature α-amylase, low-temperature liquefaction enzyme, helicase, one or more compositions in the black mold etc.
The preferred concentration of the aqueous ethanolic solution that sodium hydroxide is saturated is 80%.
Adopt technique scheme to prepare cepharanthine, adopt mixed biologic enzyme acid extraction can improve extraction efficiency, extract fully, reduce organic impurity in the crude extract, substitute the heavy method of sour molten alkali with ion exchange method and improve yield, improve cepharanthine purity, use the anion column chromatography to substitute silica gel and alumina column chromatography, reduce solvent cost, better to the xylogen removal effect.Whole technological operation is simple, organic solvent uses to lack and only uses a small amount of second alcohol and water, the operational path environmental protection, and production cost is low, is beneficial to suitability for industrialized production.
The present invention will be further described below in conjunction with embodiment, but the claimed content of the present invention is not limited to following embodiment.
Embodiment
Embodiment 1
Get and do not allow dryly piece root 1kg, the piece root is crushed to 40 orders and adds 2L water and 3ml mixed biologic enzyme liquid (pectin lyase: 50 ℃ of α low-temperature liquefaction enzymes=1:4) soaked after 1 hour, add 10 liters of 1%HCL(mass percents, lower with) aqueous solution is heated to 90 ℃ and extracted 3 hours.Separate liquid, the dregs of a decoction again add 10 liters of 1%HCL aqueous solution and decocted 3 hours, filter, merge 2 times filtrate, adopt superacicd styrene sulfonic group Zeo-karb 001X7 exchange adsorption, the enrichment cepharanthine, with adding sodium hydroxide to the 80%(mass percent, lower together) in the aqueous ethanolic solution to the solution 5L of state of saturation as elutriant, wash-out, flow velocity is 10ml/min, collect cepharanthine section elutriant and regulate pH value to 7, concentration and recovery ethanol filters, throw out with anhydrous alcohol solution after, adopt negatively charged ion DESE cellulose column purifying, collect elutriant, concentrated, dry that pale yellow powder 9.0 restrains content 99.6% with 50% ethanol water crystallization.
Embodiment 2
Get and do not allow dryly piece root 1kg, the piece root is crushed to 60 orders and adds 2L water and 1ml mixed biologic enzyme liquid (helicase: 50 ℃ of black molds=3:2) soaked after 1 hour, added 11 liters of 2%HCL(mass percents, the lower with) aqueous solution and be heated to 90 ℃ of extractions 3 hours.Separate liquid, the dregs of a decoction again add 11 liters of 2%HCL aqueous solution and decocted 3 hours, filter, merge 2 times filtrate, adopt the absorption of positively charged ion CM cellulose column, the enrichment cepharanthine, with add sodium hydroxide in 70% aqueous ethanolic solution to the solution 5L of state of saturation as elutriant, wash-out, flow velocity are 10ml/min.Collect cepharanthine section elutriant and regulate pH value to 7, concentration and recovery ethanol filters, throw out with anhydrous alcohol solution after, adopt 700B resin anion(R.A) column purification, collect elutriant, concentrated, 60% ethanol water crystallization is dry that pale yellow powder 8.6 restrains content 99.2%.
Embodiment 3
Get and do not allow dryly piece root 1kg, the piece root is crushed to 40 orders and adds 3L water and 5ml mixed biologic enzyme liquid (hemicellulase: 60 ℃ of warm α-amylases among the α=1:4) soaked after 1 hour, add 9 liters of 1%HCL(mass percents, lower with) aqueous solution is heated to 90 ℃ and extracted 3 hours.Separate liquid, the dregs of a decoction again add 9 liters of 1%HCL aqueous solution and decocted 3 hours, filter, merge 2 times filtrate, adopt slightly acidic acrylic acid Zeo-karb 152 exchange adsorption alkaloids, the enrichment cepharanthine, with add sodium hydroxide in 90% aqueous ethanolic solution to the solution 5L of state of saturation as elutriant, wash-out, flow velocity are 10ml/min.Collect cepharanthine section elutriant and regulate pH value to 7, concentration and recovery ethanol filters, throw out with anhydrous alcohol solution after, adopt D941 anion column wash-out purifying, collect elutriant, concentrated, 30% ethanol water crystallization is dry that pale yellow powder 9.1 restrains content 98.9%.
Embodiment 4
Get and do not allow dryly piece root 1kg, the piece root is crushed to 40 orders and adds 2.5L water and 4ml mixed biologic enzyme liquid (pectin lyase: 45 ℃ of helicases=2:3) soaked after 1 hour, add 10 liters of 1%HCL(mass percents, lower with) aqueous solution is heated to 90 ℃ and extracted 3 hours.Separate liquid, the dregs of a decoction again add 10 liters of 1%HCL aqueous solution and decocted 3 hours, filter, merge 2 times filtrate, adopt Zeo-karb 001X7 exchange adsorption alkaloid, the enrichment cepharanthine, with add sodium hydroxide in 80% aqueous ethanolic solution to the solution 5L of state of saturation as elutriant, wash-out, flow velocity are 10ml/min.Collect cepharanthine section elutriant and regulate pH value to 7, concentration and recovery ethanol filters, throw out with anhydrous alcohol solution after, adopt D900 anion column wash-out purifying, collect elutriant, concentrated, 50% ethanol water crystallization is dry that pale yellow powder 8.9 restrains content 99.5%.

Claims (2)

1. the separating and extracting method of a cepharanthine, it is characterized in that carrying out according to the following steps: get and do not allow dryly the piece root, pulverize, per kilogram crushed material adding 2-3L water and 0.5-5ml mixed biologic enzyme vacuole are after 1 hour, and the 1-2% mass percent HCL aqueous solution that adds 9-11 L is heated to 90 ℃ and soaked 3 hours; Separate liquid, the 1-2%HCL aqueous solution that the dregs of a decoction add 9-11 L again decocted 3 hours, filter, merge 2 times filtrate, adopt the absorption of cationic exchange coloum or cationic cellulose post, the enrichment cepharanthine, with add sodium hydroxide in the aqueous ethanolic solution of 60-90% mass concentration to the solution of state of saturation as elutriant, wash-out is collected cepharanthine section elutriant, regulates pH value to 7, concentration and recovery ethanol, filter, throw out with anhydrous alcohol solution after, with anion column wash-out purifying, collect cepharanthine section elutriant, concentrated, crystallization is drying to obtain cepharanthine; Described mixed biologic enzyme liquid is by hemicellulase, pectin lyase, middle temperature α-amylase, low-temperature liquefaction enzyme, helicase, one or more compositions in the black mold.
2. cepharanthine separating and extracting method according to claim 1, it is characterized in that with add sodium hydroxide in 80% aqueous ethanolic solution to the solution of state of saturation as elutriant.
CN 201110056911 2011-03-10 2011-03-10 Method for separating and extracting cepharanthine Active CN102146083B (en)

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Publication number Priority date Publication date Assignee Title
CN102631414B (en) * 2012-04-25 2013-09-25 成都煜泉绿健科技有限公司 SepHaniadelavayi Diels total alkaloid extraction and purification technology
CN104031058A (en) * 2014-06-30 2014-09-10 施佩蓓 Production method of cepharanthin
CN106167493B (en) * 2016-07-06 2018-06-05 河北大学 The preparation method of new cepharanthine and its application on drug

Citations (2)

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Publication number Priority date Publication date Assignee Title
CN101704821A (en) * 2009-10-20 2010-05-12 苏州派腾生物医药科技有限公司 Method for preparing stephanine
CN101891750A (en) * 2010-05-06 2010-11-24 中国科学院昆明植物研究所 Preparation method of stephanine and hydrochloride thereof

Patent Citations (2)

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Publication number Priority date Publication date Assignee Title
CN101704821A (en) * 2009-10-20 2010-05-12 苏州派腾生物医药科技有限公司 Method for preparing stephanine
CN101891750A (en) * 2010-05-06 2010-11-24 中国科学院昆明植物研究所 Preparation method of stephanine and hydrochloride thereof

Non-Patent Citations (2)

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黄加鑫等.千金藤属生物碱的研究I. 地不容中生物碱的分离与鉴定.《药学学报》.1979,第14卷(第10期),612-616. *

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