CN102234245A - Method for preparing sulforaphane - Google Patents

Method for preparing sulforaphane Download PDF

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Publication number
CN102234245A
CN102234245A CN2011101175103A CN201110117510A CN102234245A CN 102234245 A CN102234245 A CN 102234245A CN 2011101175103 A CN2011101175103 A CN 2011101175103A CN 201110117510 A CN201110117510 A CN 201110117510A CN 102234245 A CN102234245 A CN 102234245A
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Prior art keywords
liquid
add
solution
extracting
raphanin
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CN2011101175103A
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Chinese (zh)
Inventor
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 CN2011101175103A priority Critical patent/CN102234245A/en
Publication of CN102234245A publication Critical patent/CN102234245A/en
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Abstract

The invention belongs to the field of natural product separation and discloses a method for preparing sulforaphane, which comprises the following steps of: 1) crushing a radish seed medicinal material, adding water in an amount which is 2 to 3 times that of the crushed medicinal material, regulating the pH to be 4-5, performing enzymolysis at the temperature of between 45 and 50DEG C for 30 to 60 minutes, quickly heating to 80DEG C for deactivating enzymes, adding water in an amount of 5 to 6 times that of the solution, stirring, extracting, filtering out liquid, adding water in an amount which is 5 to 6 times that of the materials, extracting, mixing liquid in two times, and filtering to obtain extracting solution; 2) adsorbing the extracting solution by using macroporous resin, washing with water to reach neutrality, eluting by using 50- to 70 percent of ethanol solution in an amount which is 5 to 6 times that of the solution, collecting eluent, performing nanofiltration by using a nanofiltration membrane, and concentrating the passing liquid by using a reverse osmosis membrane to obtain concentrated solution; and 3) extracting the concentrated solution by using dichloromethane, and recovering reagents from extracting solution under reduced pressure to obtain the sulforaphane. The sulforaphane is prepared by using the macroporous resin and a membrane separation technology; and the process is easy to operate, the production cost is low and the method is suitable for industrial production.

Description

A kind of preparation method of raphanin
Technical field:
The invention belongs to the Separation of Natural Products field, particularly relate to a kind of preparation method of raphanin.
Background technology:
Raphanin (1-isothiocyanic acid-4-methylsulfonyl butane) is sulforaphen, radish thionin again; a kind of isothiocyanate that to be glucose Semen Raphani glycosides obtain through Sinapis nigra sulphur neuraminidase enzymolysis or acid hydrolysis; water-soluble, methyl alcohol and chloroform; be soluble in ether; be dissolved in sherwood oil hardly; molecular weight 177.29, molecular formula C 6H 11S 2NO, molecular structural formula:
Raphanin normal temperature is liquid state, 125~130 °/0.015mm of boiling point, [α] 19/D-107 ° (C=1.37, chloroform), [α] 14/D-136 ° (C=1.38, ethanol) down.Proof raphanins such as Talalay have good anti-cancer activity to esophagus cancer, colorectal carcinoma, mammary cancer etc., have caused the extensive concern of various countries scholar to cress sulphur glycosides composition.
Existing extraction Semen Raphani have two kinds of approach, and the one, from cross section plant, extract, the 2nd, chemosynthesis.
Patent (application number: 200910098197.6) " a kind of extracting method of sulforaphen ", the method for this patent disclosure is a raw material with exsiccant broccoli seed or fresh Caulis et Folium Brassicae capitatae ball, after the pulverizing, add entry, regulate PH4.1~4.5, add thick mustard seed sulphur thuja acid enzyme after, hydrolysis; 2. with the Caulis et Folium Brassicae capitatae lyophilize powdered after the hydrolysis; 3. in powder, add pure acetone, lixiviate is filtered, is merged acetone extraction liquid, and the filtrate vacuum concentration is become the sulforaphen crude product; 4. this sulforaphen crude product gradient elution is collected cut, vacuum concentration; With sulforaphen earlier with behind the dissolve with methanol, behind filtering with microporous membrane,, adopt the methanol-water gradient elution with C18 silica gel dress post, collect the sulforaphen cut, vacuum-drying, sulforaphen.
Patent (application number: 200910037363.1) " a kind of method of from the Caulis et Folium Brassicae capitatae bud seedling vegetable, extracting multi-functional sulforaphane ", the method of this patent disclosure be get the growth 6~10 days fresh Caulis et Folium Brassicae capitatae bud seedling vegetable be raw material, after lyophilize, pulverized 100 mesh sieves; Take by weighing raw material 100 mass parts after treatment then, add deionized water, methylene dichloride, the VC of 0.001~0.003 mass parts, the Na of 0.1~0.3 mass parts 2S, regulating pH value was 4~6,15~35 ℃ of temperature hydrolysis 6~10 hours; At last after filtration, wash, purify sulforaphane.
The clear document of delivering " purification of sulphur glycosides enzymolysis product sulforaphen and antineoplastic in vitro tests research in the broccoli seed " that waits of Shen Lian, the method that the document adopts is to be solvent with acetone, supersound extraction sulforaphen study from sulphur glycosides enzymolysis product, separate purification by normal phase silicagel column with SepHadex LH-20 gel column successively again, obtain sulforaphen.
" the synthesizing of cancer therapy drug raphanin " that Zhang Xiaolin etc. deliver, the method chemosynthesis that the document adopts.
Because raphanin is all destroyed easily in high temperature or strong acid or strong alkali environment, all more complicated is loaded down with trivial details to cause existing technology, difficult realization industrialization.
Summary of the invention
The objective of the invention is provides a kind of preparation method of raphanin in order to overcome the deficiencies in the prior art, simple for process, is fit to suitability for industrialized production.
The technology of the present invention solution is as follows:
A kind of preparation method of raphanin is characterized in that may further comprise the steps:
1) gets the Semen Raphani pulverizing medicinal materials, add 2-3 times of water gaging, regulate pH4-5,45-50 ℃ of enzymolysis 30-60min, be warming up to 80 ℃ of enzymes that go out rapidly, add 5-6 times of water gaging again and stir extraction, leach liquid, add 5-6 times of water gaging again and extract, merge liquid twice, filter extracting solution;
2) said extracted liquid adds macroporous resin adsorption, and washing is neutral, gets 5-6 and doubly measures 50-70% ethanolic soln wash-out, collects elutriant, adds the nanofiltration membrane nanofiltration, adds reverse osmosis membrane again and concentrates, and obtains concentrated solution;
3) above-mentioned concentrated solution adds dichloromethane extraction, and extraction liquid reclaim under reduced pressure reagent promptly gets raphanin.
A kind of among the optional D101 of described macroporous resin model, AB-8, HPD100 and the HZ816.Preferred AB-8 macroporous resin.
Described nanofiltration membrane is a molecular weight cut-off greater than 300 hollow cellulose film, and reverse osmosis membrane is the hollow composite membrane.
Advantage one of the present invention: the process using macroporous resin enrichment, improved the content and the purity of raphanin in the liquid, reduce the consumption of organic reagent, be more suitable for suitability for industrialized production.
Advantage two of the present invention: the process using membrane separation technique is a kind of physical separation method, selects different apertures to hold back according to molecular weight of material, neither destroys material composition, has again than highly selective, is fit to the separation and purification of raphanin, and industrialization degree is high.
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:
Embodiment 1
The Semen Raphani pulverizing medicinal materials, get 20kg, add extractor, add 40kg aqueous hydrochloric acid (pH5), be heated to 50 ℃ of enzymolysis 60min, be warming up to 80 ℃ rapidly, after 30 minutes, add 120kg water again and stir extraction 2 hours, leach liquid, add the 100kg water extraction again, merge liquid twice, filter adding AB-8 macroporous resin adsorption, washing is neutral, get 6 times of amount 50% ethanolic soln wash-outs, collect elutriant, add the hollow cellulose film nanofiltration of molecular weight cut-off 400, add the suitable quantity of water dialysis, collection adds reverse osmosis membrane again by liquid and concentrates, collect concentrated solution and add isopyknic dichloromethane extraction, extract 4 times, collect extraction liquid reclaim under reduced pressure reagent, 9g oily raphanin is through high performance liquid phase detection level 75.6%.
Embodiment 2
The Semen Raphani pulverizing medicinal materials, get 20kg, add extractor, add 60kg aqueous sulfuric acid (pH4), be heated to 45 ℃ of enzymolysis 60min, be warming up to 80 ℃ rapidly, after 30 minutes, add 100kg water again and stir extraction 2 hours, leach liquid, add the 100kg water extraction again, merge liquid twice, filter adding D101 macroporous resin adsorption, washing is neutral, get 5 times of amount 70% ethanolic soln wash-outs, collect elutriant, add the hollow cellulose film nanofiltration of molecular weight cut-off 500, add the suitable quantity of water dialysis, collection adds reverse osmosis membrane again by liquid and concentrates, collect the dichloromethane extraction that concentrated solution adds 2 times of volumes, extract 3 times, collect extraction liquid reclaim under reduced pressure reagent, 11g oily raphanin is through high performance liquid phase detection level 70.1%.
Embodiment 3:
The Semen Raphani pulverizing medicinal materials, get 20kg, add extractor, add 60kg aqueous sulfuric acid (pH4), be heated to 45 ℃ of enzymolysis 60min, be warming up to 80 ℃ rapidly, after 30 minutes, add 100kg water again and stir extraction 2 hours, leach liquid, add the 100kg water extraction again, merge liquid twice, filter adding HPD100 macroporous resin adsorption, washing is neutral, get 5 times of amount 60% ethanolic soln wash-outs, collect elutriant, add the hollow cellulose film nanofiltration of molecular weight cut-off 400, add the suitable quantity of water dialysis, collection adds reverse osmosis membrane again by liquid and concentrates, collect the dichloromethane extraction that concentrated solution adds 2 times of volumes, extract 3 times, collect extraction liquid reclaim under reduced pressure reagent, 6g oily raphanin is through high performance liquid phase detection level 71.2%.
Embodiment 4:
The Semen Raphani pulverizing medicinal materials, get 20kg, add extractor, add 60kg aqueous citric acid solution (pH4), be heated to 50 ℃ of enzymolysis 30min, be warming up to 80 ℃ rapidly, after 30 minutes, add 100kg water again and stir extraction 2 hours, leach liquid, add the 100kg water extraction again, merge liquid twice, filter adding HZ816 macroporous resin adsorption, washing is neutral, get 5 times of amount 60% ethanolic soln wash-outs, collect elutriant, add the hollow cellulose film nanofiltration of molecular weight cut-off 400, add the suitable quantity of water dialysis, collection adds reverse osmosis membrane again by liquid and concentrates, collect the dichloromethane extraction that concentrated solution adds 2 times of volumes, extract 3 times, collect extraction liquid reclaim under reduced pressure reagent, 8g oily raphanin is through high performance liquid phase detection level 73.5%.

Claims (3)

1.-plant the preparation method of raphanin, it is characterized in that may further comprise the steps:
1) gets the Semen Raphani pulverizing medicinal materials, add 2-3 times of water gaging, regulate pH4-5,45-50 ℃ of enzymolysis 30-60min, be warming up to 80 ℃ of enzymes that go out rapidly, add 5-6 times of water gaging again and stir extraction, leach liquid, add 5-6 times of water gaging again and extract, merge liquid twice, filter extracting solution;
2) said extracted liquid adds macroporous resin adsorption, and washing is neutral, gets 5-6 and doubly measures 50-70% ethanolic soln wash-out, collects elutriant, adds the nanofiltration membrane nanofiltration, adds reverse osmosis membrane again by liquid and concentrates, and obtains concentrated solution;
3) above-mentioned concentrated solution adds dichloromethane extraction, and extraction liquid reclaim under reduced pressure reagent promptly gets raphanin.
2. according to the preparation method of the described raphanin of claim 1, it is characterized in that step 2) a kind of among the optional D101 of described macroporous resin model, AB-8, HPD100 and the HZ816.
3. according to the preparation method of the described raphanin of claim 1, it is characterized in that step 2) described nanofiltration membrane is molecular weight cut-off greater than 300 hollow cellulose film, reverse osmosis membrane is the hollow composite membrane.
CN2011101175103A 2011-05-06 2011-05-06 Method for preparing sulforaphane Pending CN102234245A (en)

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103272033A (en) * 2013-04-16 2013-09-04 阚兆云 Traditional Chinese medicine extract and preparation method thereof
CN103349698A (en) * 2013-07-20 2013-10-16 石强 Natural pharmaceutical extractive, and preparation method thereof
CN103393759A (en) * 2013-07-20 2013-11-20 石强 Natural pharmaceutical composition and preparation method thereof
CN103393755A (en) * 2013-07-20 2013-11-20 石强 Natural medicine extract as well as preparation method and application thereof
CN104083337A (en) * 2014-05-22 2014-10-08 浙江大学苏州工业技术研究院 Preparation method of sulforaphane capsule
CN104402787A (en) * 2014-10-24 2015-03-11 苏州泽达兴邦医药科技有限公司 Method for separation and purification of sulforaphane
CN106349133A (en) * 2016-08-24 2017-01-25 徐金蝶 Extraction method of sulforaphane
CN106631946A (en) * 2016-09-21 2017-05-10 广州六顺生物科技有限公司 Method for preparing sulforaphene

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103272033A (en) * 2013-04-16 2013-09-04 阚兆云 Traditional Chinese medicine extract and preparation method thereof
CN103349698A (en) * 2013-07-20 2013-10-16 石强 Natural pharmaceutical extractive, and preparation method thereof
CN103393759A (en) * 2013-07-20 2013-11-20 石强 Natural pharmaceutical composition and preparation method thereof
CN103393755A (en) * 2013-07-20 2013-11-20 石强 Natural medicine extract as well as preparation method and application thereof
CN104083337A (en) * 2014-05-22 2014-10-08 浙江大学苏州工业技术研究院 Preparation method of sulforaphane capsule
CN104402787A (en) * 2014-10-24 2015-03-11 苏州泽达兴邦医药科技有限公司 Method for separation and purification of sulforaphane
CN106349133A (en) * 2016-08-24 2017-01-25 徐金蝶 Extraction method of sulforaphane
CN106631946A (en) * 2016-09-21 2017-05-10 广州六顺生物科技有限公司 Method for preparing sulforaphene

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Application publication date: 20111109