CN1563104A - Method for extracting and purifying polysaccharide from aconite root of Chinese traditional medicine - Google Patents
Method for extracting and purifying polysaccharide from aconite root of Chinese traditional medicine Download PDFInfo
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- CN1563104A CN1563104A CN 200410026912 CN200410026912A CN1563104A CN 1563104 A CN1563104 A CN 1563104A CN 200410026912 CN200410026912 CN 200410026912 CN 200410026912 A CN200410026912 A CN 200410026912A CN 1563104 A CN1563104 A CN 1563104A
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Abstract
The invention relates to a preparation method of Chinese medicine aconite polysaccharide. Said method uses black sliced aconite as raw material, and adopts the following steps: sorting, removing impurity, grinding by using grinder, extracting by using ultrasoic extractor, centrifugalizing extract, filtering, concentrating, adding ethyl alcohol to make it concentration be up to 75%, fully-stirring, standing still, centrifugally-filtering to obtain crude aconite polysaccharide, redissolving said crude polysaccharide in distilled water, removing impurity, secondary concentrating, alcohol precipitation, washing by using absolute ethyl alcohol, acetone and ethyl ether successively and drying so as to obtain the purified aconite polysaccharide refined product. It has the actions of reducing blood glucose of mouse and raising immunological function of mouse.
Description
The present invention relates to a kind of from the Chinese medicine monkshood method of extraction separation purified polysaccharide, belong to the pharmaceutical chemistry field.
In recent years to having the polysaccharide material of physiologically active, through pharmacological research and clinical application, to confirm to have hypoglycemic, the effect of antitumor and raise immunity is subjected to increasing expert's attention.The present both at home and abroad polysaccharide material that extracts from Chinese medicine for panaxan, Radix Et Caulis Acanthopanacis Senticosi polysaccharide, polyporusum bellatus, lycium barbarum polysaccharide, lentinan etc. has been applied to clinically, obtains gratifying effect.Monkshood belongs to the lateral root of Ranunculaceae Aconitum (Aconitum Carmichaeli Debx) plant, is a kind of Chinese medicine commonly used, has the function of recuperating deplered YANG and rescuing the patient from collapse, warming and recuperating the kidney-YANG.For the research of effective constituent in the monkshood, mainly concentrate on the Radix Aconiti Lateralis Preparata alkaloid aspect, then seldom both at home and abroad to STUDY ON POLYSACHAROSE in the monkshood.The extraction separation purification process of polysaccharide material has multiple, through consulting document, provided by the invention in the extraction separation purification process of homemade monkshood (Radix Aconiti Lateralis Preparata) polysaccharide do not see that report is arranged.The contriver carries out the experiment of immunoregulation effect and hypoglycemic activity repeatedly when its efficient part of research screening, proved conclusively its drug action.
The purpose of this invention is to provide three kinds from Chinese medicine monkshood rhizome processed product---extract the method for crude monkshood polysaccharide the Radix Aconiti Lateralis Preparata.
Another object of the present invention provides the purification process of above-mentioned Crude polysaccharides.
The present invention extracts crude monkshood polysaccharide from the Chinese medicine monkshood method comprises the steps:
1. with Chinese medicine monkshood Radix Aconiti Lateralis Preparata sorting removal of impurities, pulverize, cross the 20-50 mesh sieve.
2. extract.With one of following three kinds of methods.
(2.1) place the supersound extraction device, adding pH value is 8~10 the NaOH aqueous solution, and 80~100 ℃ of temperature are extracted three times, extract (solid-to-liquid ratio is 1: 10~20) 0.5~2 hour for the first time; Filter residue extracts secondary (solid-to-liquid ratio is 1: 5~10), each 0.5~1.5 hour again.The dregs of a decoction can be used for preparing Radix Aconiti Lateralis Preparata alkaloid.
(2.2) adding pH value is 8~10 the NaOH aqueous solution, boils earlier 1~2 hour, changes in the supersound extraction device again and extracts, and 80~100 ℃, 0.5~1.5 hour, uses filtered through gauze; Filter residue is supersound extraction 1~2 time (solid-to-liquid ratio is 1: 5~10) again, 0.5~1.5 hour/time.The dregs of a decoction can be used for preparing Radix Aconiti Lateralis Preparata alkaloid.
(2.3) carry total alkaloids with Chinese medicine monkshood Radix Aconiti Lateralis Preparata sorting removal of impurities, pulverize, get 50g and add 95% ethanol 800ml with pulverizer, be transferred in the apparatus,Soxhlet's, 80~90 ℃ of ultrasonic refluxing extraction 2 hours are filtered, ethanol extract concentrates, and is used to prepare Radix Aconiti Lateralis Preparata alkaloid, reclaims ethanol.It is 8~10 NaOH aqueous solution supersound extraction 2~3 times that elder generation's supersound extraction Radix Aconiti Lateralis Preparata total alkaloids, the dregs of a decoction add pH value again, extraction (solid-to-liquid ratio is 1: 10~20) for the first time 0.5~2 hour; Filter residue extracts (solid-to-liquid ratio is 1: 5~10), 0.5~1.5 hour/time again 1~2 time.
3. filtered through gauze, united extraction liquid, place while hot whizzer centrifugal (2000~4000r/min) 15~25 minutes, filter.
4. filtrate is concentrated into 1/3~1/5 volume.
5. the adding dehydrated alcohol makes alcohol concn reach 70~85%, fully stirs, and leaves standstill, and gets the Crude polysaccharides precipitation.
6. centrifugal, filtration, dry Crude polysaccharides.
The crude monkshood polysaccharide purification process:
1. Crude polysaccharides redissolves with 1-2 times of (w/w) distilled water.
2. add phenol/chloroform (1: 1 v/v) reagent by 3: 1~5: 1 (v/v) volumes, abundant stirring and evenly mixing, standing demix.
3. centrifugal (2000~3000r/min) 25 minutes, filter to get filtrate.
4. after filtrate is repeated removal of impurities 2~4 times, concentrating under reduced pressure, alcohol precipitation, centrifugal, polysaccharide precipitation.
Use dehydrated alcohol, acetone, the ether thorough washing 2~3 times of 3~5 times of volumes successively, after the vacuum-drying, pulverize refining Radix Aconiti Lateralis Preparata polysaccharide.
The invention has the advantages that: the using ultrasound extractive technique, improve impurity removal process, shortened extraction time greatly, yield height, purity height, cost are low.By pharmacodynamic experiment, prove to have immunoloregulation function and hypoglycemic activity.
Provide embodiment below:
[example 1]:
(1) supersound extraction is pulverized Chinese medicine monkshood Radix Aconiti Lateralis Preparata sorting removal of impurities with pulverizer, gets 50g; The NaOH aqueous solution 750ml that adds pH=9, temperature is under 90 ℃ the condition, the supersound extraction secondary.Extracted (solid-to-liquid ratio is 1: 15) 1 hour for the first time; Filter residue extracts secondary (solid-to-liquid ratio is 1: 5) again, and each 0.5 hour, with four layers of filtered through gauze, No. three extracting solutions are merged, placed whizzer centrifugal (3000r/min) while hot 20 minutes, filter to get filtrate.The dregs of a decoction are used to prepare Radix Aconiti Lateralis Preparata alkaloid.
(2) concentrate, the alcohol precipitation filtrate decompression is concentrated into 1/5 volume, adds dehydrated alcohol, makes the ethanol final concentration reach 75%, leaves standstill 4 hours, makes polysaccharide precipitation complete, places whizzer centrifugal (3000r/min) 10 minutes, filter polysaccharide precipitation.
(3) redissolution, removal of impurities are dissolved in the gained Crude polysaccharides in 2 times of volume distilled water again, add phenol/chloroform (1: 1 v/v) reagent by 5: 1 (v/v) volumes, abundant stirring and evenly mixing, and behind the standing demix, centrifugal (3000r/min) 25 minutes filters to get filtrate.Filtrate is repeated (2), the removal of impurities of (3) step secondary.
(4) concentrate, the filtrate after the alcohol precipitation removal of impurities, through being evaporated to 1/2 volume, add dehydrated alcohol, make final concentration reach 75%, left standstill 4 hours, make polysaccharide precipitation complete, polysaccharide precipitation was placed whizzer centrifugal (3000r/min) 20 minutes, filter polysaccharide precipitation.
(5) refining with the polysaccharide precipitation behind the secondary alcohol precipitation, successively use 5 times of volumes dehydrated alcohol, acetone, ether thorough washing each twice, Radix Aconiti Lateralis Preparata polysaccharide.
(6) drying will be made with extra care polysaccharide and be moved to vacuum drying oven.
(7) dried polysaccharide is pulverized, got white powder Radix Aconiti Lateralis Preparata polysaccharide elaboration 6.2g, the encloses container of packing into is moistureproof to be preserved, standby.
[example 2]:
(1) it is ultrasonic with Chinese medicine monkshood Radix Aconiti Lateralis Preparata sorting removal of impurities that first water is carried the back, pulverizes with pulverizer, gets 50g; Add the NaOH aqueous solution 750ml (solid-to-liquid ratio is 1: 15) of pH=9,100 ℃ were boiled 0.5 hour, the supersound extraction of migrating, and 90 ℃ were extracted filtered through gauze 1 hour; Filter residue extracts 1 hour (solid-to-liquid ratio is 1: 5) with ultrasonic 90 ℃ again, with four layers of filtered through gauze, secondary raffinate is merged, and places whizzer centrifugal (3000r/min) while hot 20 minutes, filters to get filtrate.The dregs of a decoction are used to prepare Radix Aconiti Lateralis Preparata alkaloid.
(2) concentrate, the alcohol precipitation filtrate decompression is concentrated into 1/5 volume, adds dehydrated alcohol, makes the ethanol final concentration reach 75%, leaves standstill 4 hours, makes polysaccharide precipitation complete, places whizzer centrifugal (3000r/min) 10 minutes, filter polysaccharide precipitation.
(3) redissolve, removal of impurities is dissolved in the gained Crude polysaccharides in 2 times of volume distilled water again, by 5: 1 (v/v) volumes add phenol/chloroform (1: 1v/v) reagent, abundant stirring and evenly mixing, behind the standing demix, centrifugal (3000r/min) 25 minutes filters to get filtrate.Filtrate is repeated (2), the removal of impurities of (3) step secondary.
(4) concentrate, the filtrate after the alcohol precipitation removal of impurities, through being evaporated to 1/2 volume, add dehydrated alcohol, make final concentration reach 75%, left standstill 4 hours, make polysaccharide precipitation complete, polysaccharide precipitation was placed whizzer centrifugal (3000r/min) 20 minutes, filter polysaccharide precipitation.
(5) refining with the polysaccharide precipitation behind the secondary alcohol precipitation, successively use 5 times of volumes dehydrated alcohol, acetone, ether thorough washing each twice, Radix Aconiti Lateralis Preparata polysaccharide.
(6) drying will be made with extra care polysaccharide and be moved to vacuum drying oven.
(7) dried polysaccharide is pulverized, got white powder Radix Aconiti Lateralis Preparata polysaccharide elaboration 6.7g, the encloses container of packing into is moistureproof to be preserved, standby.
[example 3]:
(1) carries total alkaloids with Chinese medicine monkshood Radix Aconiti Lateralis Preparata sorting removal of impurities, pulverize, get 50g and add 95% ethanol 800ml with pulverizer, be transferred in the apparatus,Soxhlet's, 80~90 ℃ of ultrasonic refluxing extraction 2 hours are filtered, ethanol extract concentrates, and is used to prepare Radix Aconiti Lateralis Preparata alkaloid, reclaims ethanol.
(2) to add pH value be 8~10 NaOH aqueous solution supersound extraction 2 times to the supersound extraction dregs of a decoction, and add 750ml the first time, extracted four layers of filtered through gauze 1 hour; Filter residue added the 500mlNaOH extraction with aqueous solution 1 hour again, and four layers of filtered through gauze merge extracted twice liquid, place whizzer centrifugal (3000r/min) while hot 20 minutes, filter to get filtrate.
(3) concentrate, the alcohol precipitation filtrate decompression is concentrated into 1/5 volume, adds dehydrated alcohol, makes the ethanol final concentration reach 75%, leaves standstill 4 hours, makes polysaccharide precipitation complete, places whizzer centrifugal (3000r/min) 10 minutes, filter polysaccharide precipitation.
(4) redissolution, removal of impurities are dissolved in the gained Crude polysaccharides in 2 times of volume distilled water again, add phenol/chloroform (1: 1 v/v) reagent by 5: 1 (v/v) volumes, abundant stirring and evenly mixing, and behind the standing demix, centrifugal (3000r/min) 25 minutes filters to get filtrate.Filtrate is repeated (2), the removal of impurities of (3) step secondary.
(5) concentrate, the filtrate after the alcohol precipitation removal of impurities, through being evaporated to 1/2 volume, add dehydrated alcohol, make final concentration reach 75%, left standstill 4 hours, make polysaccharide precipitation complete, polysaccharide precipitation was placed whizzer centrifugal (3000r/min) 20 minutes, filter polysaccharide precipitation.
(6) refining with the polysaccharide precipitation behind the secondary alcohol precipitation, successively use 5 times of volumes dehydrated alcohol, acetone, ether thorough washing each twice, Radix Aconiti Lateralis Preparata polysaccharide.
(7) drying will be made with extra care polysaccharide and be moved to vacuum drying oven.
(8) dried polysaccharide is pulverized, got white powder Radix Aconiti Lateralis Preparata polysaccharide elaboration 6.0g, the encloses container of packing into is moistureproof to be preserved, standby.
Claims (2)
1. the extracting method of polysaccharide in the Chinese medicine monkshood is characterized in that comprising the steps:
(1) with Chinese medicine monkshood Radix Aconiti Lateralis Preparata sorting removal of impurities, pulverize, cross the 20-50 mesh sieve;
(2) extract
(2.1) place the supersound extraction device, adding pH value is 8~10 the NaOH aqueous solution, and 80~100 ℃ of temperature are extracted three times, extract (solid-to-liquid ratio is 1: 10~20) 0.5~2 hour for the first time; Filter residue extracts secondary (solid-to-liquid ratio is 1: 5~10) again, and each 0.5~1.5 hour, or
(2.2) adding pH value is 8~10 the NaOH aqueous solution, boils earlier 1~2 hour, changes in the supersound extraction device again and extracts, and 80~100 ℃, 0.5~1.5 hour, uses filtered through gauze; Filter residue is supersound extraction 1~2 time (solid-to-liquid ratio is 1: 5~10) again, and 0.5~1.5 hour/time, or
(2.3) carry total alkaloids with Chinese medicine monkshood Radix Aconiti Lateralis Preparata sorting removal of impurities, pulverize, get 50g and add 95% ethanol 800ml with pulverizer, be transferred in the apparatus,Soxhlet's, 80~90 ℃ of ultrasonic refluxing extraction 2 hours are filtered, ethanol extract concentrates, and is used to prepare Radix Aconiti Lateralis Preparata alkaloid, reclaims ethanol; It is 8~10 NaOH aqueous solution supersound extraction 2~3 times that elder generation's supersound extraction Radix Aconiti Lateralis Preparata total alkaloids, the dregs of a decoction add pH value again, extraction (solid-to-liquid ratio is 1: 10~20) for the first time 0.5~2 hour; Filter residue extracts (solid-to-liquid ratio is 1: 5~10), 0.5~1.5 hour/time again 1~2 time;
(3) filtered through gauze, united extraction liquid, place while hot whizzer centrifugal (2000~4000r/min) 15~25 minutes, filter;
(4) filtrate is concentrated into 1/3~1/5 volume;
(5) add dehydrated alcohol, make alcohol concn reach 70~85%, fully stir, leave standstill, get the Crude polysaccharides precipitation; Centrifugal, filtration, dry Crude polysaccharides.
2. the purification process of polysaccharide in the Chinese medicine monkshood is characterized in that comprising the steps:
(1) Crude polysaccharides redissolves with 1-2 times of (w/w) distilled water;
(2) add phenol/chloroform (1: 1v/v) reagent, abundant stirring and evenly mixing, standing demix by 3: 1~5: 1 (v/v) volumes;
(3) centrifugal (2000~3000r/min) 25 minutes, filter to get filtrate;
(4) after filtrate is repeated removal of impurities 2~4 times, concentrating under reduced pressure, alcohol precipitation, centrifugal, polysaccharide precipitation; Use dehydrated alcohol, acetone, the ether thorough washing 2~3 times of 3~5 times of volumes successively, after the vacuum-drying, pulverize refining Radix Aconiti Lateralis Preparata polysaccharide.
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Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
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CN101839863A (en) * | 2010-02-10 | 2010-09-22 | 宁波御坊堂生物科技有限公司 | Extraction and determination method of crude polysaccharide in ganoderma spirulina |
CN101538334B (en) * | 2009-04-24 | 2012-05-30 | 昆明理工大学 | Method for extracting medicinal large leaf moss polysaccharide |
CN103181957A (en) * | 2013-01-15 | 2013-07-03 | 浙江中医药大学 | Preparation technology capable of completely removing acute toxicity of prepared aconite roots and keeping pharmacological activity of prepared aconite roots |
CN104387481A (en) * | 2014-10-22 | 2015-03-04 | 苏州市中医院 | Extracting method for ripe monkshood polyose and novel application thereof |
CN106492002A (en) * | 2016-10-17 | 2017-03-15 | 紫云自治县紫香源农林科技有限责任公司 | A kind of extracting method of Germinatus Bambusae sinospinosae liquid |
CN109021127A (en) * | 2018-06-29 | 2018-12-18 | 北京工商大学 | A kind of extracting method of Polysaccharide from Portulaca oleracea |
CN110746512A (en) * | 2018-07-24 | 2020-02-04 | 广东省中医院 | Radix aconiti carmichaeli uniform polysaccharide, and preparation method and application thereof |
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2004
- 2004-04-19 CN CN 200410026912 patent/CN1563104A/en active Pending
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101538334B (en) * | 2009-04-24 | 2012-05-30 | 昆明理工大学 | Method for extracting medicinal large leaf moss polysaccharide |
CN101839863A (en) * | 2010-02-10 | 2010-09-22 | 宁波御坊堂生物科技有限公司 | Extraction and determination method of crude polysaccharide in ganoderma spirulina |
CN101839863B (en) * | 2010-02-10 | 2012-03-21 | 宁波御坊堂生物科技有限公司 | Extraction and determination method of crude polysaccharide in ganoderma spirulina |
CN103181957A (en) * | 2013-01-15 | 2013-07-03 | 浙江中医药大学 | Preparation technology capable of completely removing acute toxicity of prepared aconite roots and keeping pharmacological activity of prepared aconite roots |
CN104387481A (en) * | 2014-10-22 | 2015-03-04 | 苏州市中医院 | Extracting method for ripe monkshood polyose and novel application thereof |
CN106492002A (en) * | 2016-10-17 | 2017-03-15 | 紫云自治县紫香源农林科技有限责任公司 | A kind of extracting method of Germinatus Bambusae sinospinosae liquid |
CN109021127A (en) * | 2018-06-29 | 2018-12-18 | 北京工商大学 | A kind of extracting method of Polysaccharide from Portulaca oleracea |
CN109021127B (en) * | 2018-06-29 | 2021-09-10 | 北京工商大学 | Extraction method of purslane polysaccharide |
CN110746512A (en) * | 2018-07-24 | 2020-02-04 | 广东省中医院 | Radix aconiti carmichaeli uniform polysaccharide, and preparation method and application thereof |
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