CN102477104A - Method for separating and purifying polysaccharide from Hovenia acerba - Google Patents
Method for separating and purifying polysaccharide from Hovenia acerba Download PDFInfo
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- CN102477104A CN102477104A CN2010105577500A CN201010557750A CN102477104A CN 102477104 A CN102477104 A CN 102477104A CN 2010105577500 A CN2010105577500 A CN 2010105577500A CN 201010557750 A CN201010557750 A CN 201010557750A CN 102477104 A CN102477104 A CN 102477104A
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- polysaccharide
- raisin tree
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Abstract
The present invention discloses a method for separating and purifying polysaccharide from Hovenia acerba. The method is characterized by pulverizing raw material, leaching with hot water, filtering, and concentrating to obtain an extract; adding alcohol into the extract for precipitation, and centrifuging to obtain crude polysaccharide of Hovenia acerba; and dissolving the crude polysaccharide in an appropriate amount of hot water, performing chromatography via a diethylaminoethyl (DEAE) cellulose column for removing impurity, decolorizing via an ion exchange resin column, dialyzing via a semi-permeable membrane, and drying. The prepared polysaccharide is clear white, and has little impurity and high purity. In addition, the decolorization step employs ion exchange resin, to achieve good decolorization effect and high retention rate of polysaccharide.
Description
Technical field:
The present invention relates to a kind of separation purification method of honey raisin tree polysaccharide, belong to technical field of traditional Chinese medicines.
Background technology:
Calligonum (Hovenia? Cerba? Lindl.) Is Rhamnaceae Calligonum is a deciduous tree, commonly known dulcis, hooks, chicken paws, thousands of words fruit, Papaya, Alondra.Its whole body all is precious, is the famous and precious tree of a kind of fruit, medicine, material dual-purpose, and the wide development prospect is arranged.Fruit containing glucose and calcium malate, seeds contain saponin EHD, has sedative, anticonvulsant, and lowering blood pressure and other effects.But the meat dispelling the wind and dampness pathogens has the sharp diuresis of heat-clearing to quench the thirst and falls fiery detoxicating functions, and bark can be invigorated blood circulation, the Shujin detoxifcation, and carpopodium property is sweet flat, and the effect of promoting digestion and enriching blood is arranged, and is nontoxic, cures mainly a wind, the sharp stool and urine of moistening five ZANG-organs.Xylem contains the acid of dicarboxylicacid triterpene compound trifoliate orange seed (Hobeniaaced, C
30H
80O
5).Employings such as Zhu Jiongbo anthrone-its polysaccharide content of sulfuric acid colorimetric method for determining is 13.83%, both at home and abroad effect research of honey raisin tree active polysaccharide is not appeared in the newspapers as yet at present.
Polysaccharide is the macromolecular substance in the natural organic-compound; Some active polysaccharide has antitumor and effect raise immunity; Some polysaccharide then have complicated control cell fission and differentiation, regulate many-sided functions such as cell growth and aging, the effect that the polysaccharide of modification then has anti-HIV to infect, some important physical process such as inflammatory reactions; Acceptor identification, immunoreation etc. are all closely related with polysaccharide molecule.Existing a large amount of pharmacological research shows, the polysaccharide in the vegetable polysaccharides, particularly Chinese medicine has immunomodulatory, multiple physiologically active such as antitumor, antiviral, anti-infective, hypoglycemic, and its crude drug in whole belongs to help class Chinese medicine mostly.Though a lot of synthetic chemotherapeutics are arranged at present; But they involve normal cell too in kill cancer cell; Show a lot of spinoffs clinically; Particularly chemotherapeutics causes damage to patient's immune system, but the genetoxic of plant amedica is not too obvious, shows that herbal medicine is having its special advantages and wide application prospect aspect the anticancer mutation.Therefore, the treatment of Polysaccharide Immunoregulators on tumour is very attractive.
Ion exchange resin is a kind of porousness cross-linked polymer, has higher specific surface area, thereby has good adsorptive power.Nonionic adsorption resin belongs to two phase structure: polymer phase and hole.Polymer phase is to be piled up by numerous nano grade polymer particulates to form, and is the hole between the particulate.Can be adjusted to the example size through adjustment synthetic method, post aperture and specific surface area etc., thereby made resin have the different adsorption performance.In addition, polymeric adsorbent can be divided into four kinds of nonpolar, Semi-polarity, polarity and strong polarity again by surface properties.Different surface character makes polymeric adsorbent possess different adsorption mechanism and absorption property, therefore makes resin obtain many-sided purposes.There are some researches show, replace gac commonly used to decolour, not only can accelerate filtration velocity with exchange resin, and decolorizing effect, also reach the purpose of impurity elimination simultaneously, thereby improve the quality of product.Resin has ionization simultaneously except that decolouring, can play ion exchange reaction with the ion in the solution; Add resins for universal use also have stronger ligand group (OH ,-NH3); Can play complexing action to metals ion, thereby remove the solution metal ion, play a part impurity elimination.
Existing polysaccharide decolouring technology often adopts activated carbon decolorizing, though cheap, must use Powdered Activated Carbon during operation, the fatal shortcoming that exists the charcoal residue in the solution of decolouring back to be difficult to go out fully; Ydrogen peroxide 50 decolouring need purpose product has stability, is difficult for the characteristics of oxidation; But because the ydrogen peroxide 50 decoloring method is the pigment in the direct oxidation extracting solution; Do not remove; And might be with the structure deteriorate of polysaccharide chain, the pigment after the oxidation is still stayed in the extracting solution as impurity, can not guarantee its last quality.The present invention adopts the ion exchange resin treatment ability big, and scope is wide, and percent of decolourization is high, makes the polysaccharide retention rate high, renewable recycling, and cost is lower.
Summary of the invention:
The objective of the invention is to be to provide a kind of separation purification method of honey raisin tree polysaccharide, from honey raisin tree through hot water lixiviate, alcohol precipitation, the removal of impurities of centrifugal DEAE cellulose chromatography, ion exchange resin column decolouring, semi-permeable membranes dialysis and dry finished product.This method is extracted the honey raisin tree active polysaccharide that obtains higher degree.
For achieving the above object, the present invention has adopted following technical measures:
The separation purification method of honey raisin tree polysaccharide is characterized in that comprising the following steps:
(1) polysaccharide extraction liquid preparation: honey raisin tree is pulverized, heating flooding 2-3 time, lixiviate 24-48 hour, filter, merge vat liquor, concentrated honey raisin tree medicinal extract;
(2) alcohol precipitation: the edible ethanol stirring that honey raisin tree medicinal extract is added 3-5 times of 80-90% staticly settled after 5-15 minute, and supernatant is removed in centrifuging, the collecting precipitation thing;
(3) removal of impurities, decolouring: centrifugal sediment is used the hot water stirring and dissolving, last DEAE cellulose column, 0-0.5mol/LNaCl solution gradient wash-out is gone up the ion exchange resin column adsorption bleaching again, and lower column liquid filters the back with the semi-permeable membranes dialysis, is drying to obtain finished product.
The rotating speed of whizzer is 5000-10000rpm in the said step (2).
Cellulose column adopts DEAE-23, DEAE-52 in the said step (3).
Optional phenylethylene resin series of said step (3) intermediate ion exchange resin and acrylic resin or both hybrid resins.Phenylethylene resin series can adopt Asmit 265P, Duolite A171P resin anion(R.A); Acrylic resin can adopt Asmit295, DoliteA172, IRA958S, A860 resin anion(R.A).
Semi-permeable membranes is the semi-permeable membranes dialysis tubing of molecular weight cut-off 4000-10000 in the said step (3), dialyses 24-48 hour.
Drying means can adopt a kind of in lyophilize or the spraying drying in the said step (3).
Characteristics of the present invention: 1) honey raisin tree obtains the honey raisin tree polysaccharide through hot water lixiviate, alcohol precipitation, removal of impurities, decolouring, drying and other steps, and the preparation method is simple, and is easy to implement; 2) ion-exchange resin decolorization is effective, and product purity is high; 3) for the application of honey raisin tree provides favourable technique means, the honey raisin tree polysaccharide that makes can be used for its active further research, and for honey raisin tree polysaccharide medicine and healthcare products provide big content of starting materials, has promoted the Economic development of honey raisin tree.
To combine embodiment to further specify the present invention below, but the scope that the present invention requires to protect is not limited to following embodiment.
Embodiment:
Embodiment 1:
The honey raisin tree that to dry in the shade is ground into 80 order meal, places extractor, adds 10 times of 90-100 ℃ of hot water constant temperature soaking and extracting, extract repeatedly 3 times, each 24 hours, united extraction liquid, concentrate medicinal extract; 90% edible ethanol that in medicinal extract, adds 3 times of volumes stirs and staticly settles after 5 minutes, and supernatant is removed in centrifuging; The collecting precipitation thing is used the hot water stirring and dissolving with throw out, last DEA-23 cellulose column; 0-0.5mol/LNaCl the solution gradient wash-out, the Asmit 265P anionite-exchange resin dress post that pre-treatment is good, elutriant is concentrated into 0.3kg/L; With the 2BV/h upper prop, collect lower column liquid, be concentrated into minimum volume after the filtration; With pack into the semi-permeable membranes dialysis tubing dialysis 48 hours of molecular weight cut-off 4000 of liquid concentrator, spraying drying promptly gets the honey raisin tree polysaccharide, and gained honey raisin tree purity of polysaccharide reaches about 70%.
Embodiment 2:
The honey raisin tree that to dry in the shade is ground into 100 order meal, places extractor, adds 14 times of 90-100 ℃ of hot water constant temperature soaking and extracting, extract repeatedly 2 times, each 48 hours, united extraction liquid, concentrate medicinal extract; 85% edible ethanol that in medicinal extract, adds 4 times of volumes stirs and staticly settles after 15 minutes, and supernatant is removed in centrifuging; The collecting precipitation thing is used the hot water stirring and dissolving with throw out, last DEAE-52 cellulose column; 0-0.5mol/LNaCl the solution gradient wash-out, Asmit 265P that pre-treatment is good and Asmit295 mixed anion exchange resin dress post, elutriant is concentrated into 0.44kg/L; With the 2BV/h upper prop, collect lower column liquid, be concentrated into minimum volume after the filtration; With pack into the semi-permeable membranes dialysis tubing dialysis 24 hours of molecular weight cut-off 7000 of liquid concentrator, spraying drying promptly gets the honey raisin tree polysaccharide, and gained honey raisin tree purity of polysaccharide reaches about 70%.
Embodiment 3:
The honey raisin tree that to dry in the shade is ground into 80 order meal, places extractor, adds 13 times of 90-100 ℃ of hot water constant temperature soaking and extracting, extract repeatedly 3 times, each 24 hours, united extraction liquid, concentrate medicinal extract; 90% edible ethanol that in medicinal extract, adds 4 times of volumes stirs and staticly settles after 10 minutes, and supernatant is removed in centrifuging; The collecting precipitation thing is used the hot water stirring and dissolving with throw out, last DEAE-52 cellulose column; 0-0.5mol/LNaCl the solution gradient wash-out, the Duolite A171P anionite-exchange resin dress post that pre-treatment is good, elutriant is concentrated into 0.38kg/L; With the 2BV/h upper prop, collect lower column liquid, be concentrated into minimum volume after the filtration; With pack into the semi-permeable membranes dialysis tubing dialysis 36 hours of molecular weight cut-off 4000 of liquid concentrator, lyophilize promptly gets the honey raisin tree polysaccharide, and gained honey raisin tree purity of polysaccharide reaches about 70%.
Embodiment 4:
The honey raisin tree that to dry in the shade is ground into 60 order meal, places extractor, adds 10 times of 90-100 ℃ of hot water constant temperature soaking and extracting, extract repeatedly 2 times, each 36 hours, united extraction liquid, concentrate medicinal extract; 80% edible ethanol that in medicinal extract, adds 4 times of volumes stirs and staticly settles after 15 minutes, and supernatant is removed in centrifuging; The collecting precipitation thing is used the hot water stirring and dissolving with throw out, last DEAE-52 cellulose column; 0-0.5mol/LNaCl the solution gradient wash-out, the Dolite A172 anionite-exchange resin dress post that pre-treatment is good, elutriant is concentrated into 0.5kg/L; With the 2BV/h upper prop, collect lower column liquid, be concentrated into minimum volume after the filtration; With pack into the semi-permeable membranes dialysis tubing dialysis 48 hours of molecular weight cut-off 6000 of liquid concentrator, spraying drying promptly gets the honey raisin tree polysaccharide, and gained honey raisin tree purity of polysaccharide reaches about 70%.
Embodiment 5:
The honey raisin tree that to dry in the shade is ground into 40 order meal, places extractor, adds 12 times of 90-100 ℃ of hot water constant temperature soaking and extracting, extract repeatedly 2 times, each 48 hours, united extraction liquid, concentrate medicinal extract; 90% edible ethanol that in medicinal extract, adds 3 times of volumes stirs and staticly settles after 15 minutes, and supernatant is removed in centrifuging; The collecting precipitation thing is used the hot water stirring and dissolving with throw out, last DEAE-23 cellulose column; 0-0.5mol/LNaCl the solution gradient wash-out, Duolite A171P that pre-treatment is good and Dolite A172 mixed anion exchange resin dress post, elutriant is concentrated into 0.40kg/L; With the 2BV/h upper prop, collect lower column liquid, be concentrated into minimum volume after the filtration; With pack into the semi-permeable membranes dialysis tubing dialysis 24 hours of molecular weight cut-off 5000 of liquid concentrator, lyophilize promptly gets the honey raisin tree polysaccharide, and gained honey raisin tree purity of polysaccharide reaches about 70%.
Embodiment 6:
The honey raisin tree that to dry in the shade is ground into 50 order meal, places extractor, adds 13 times of 90-100 ℃ of hot water constant temperature soaking and extracting, extract repeatedly 3 times, each 30 hours, united extraction liquid, concentrate medicinal extract; 85% edible ethanol that in medicinal extract, adds 3 times of volumes stirs and staticly settles after 10 minutes, and supernatant is removed in centrifuging; The collecting precipitation thing is used the hot water stirring and dissolving with throw out, last DEAE-23 cellulose column; 0-0.5mol/LNaCl the solution gradient wash-out, the A860 anionite-exchange resin dress post that pre-treatment is good, elutriant is concentrated into 0.34kg/L; With the 2BV/h upper prop, collect lower column liquid, be concentrated into minimum volume after the filtration; With pack into the semi-permeable membranes dialysis tubing dialysis 36 hours of molecular weight cut-off 10000 of liquid concentrator, lyophilize promptly gets the honey raisin tree polysaccharide, and gained honey raisin tree purity of polysaccharide reaches about 70%.
Embodiment 7:
The honey raisin tree that to dry in the shade is ground into 60 order meal, places extractor, adds 15 times of 90-100 ℃ of hot water constant temperature soaking and extracting, extract repeatedly 2 times, each 24 hours, united extraction liquid, concentrate medicinal extract; 80% edible ethanol that in medicinal extract, adds 4 times of volumes stirs and staticly settles after 15 minutes, and supernatant is removed in centrifuging; The collecting precipitation thing is used the hot water stirring and dissolving with throw out, last DEAE-52 cellulose column; 0-0.5mol/LNaCl the solution gradient wash-out, the IRA958S anionite-exchange resin dress post that pre-treatment is good, elutriant is concentrated into 0.35kg/L; With the 2BV/h upper prop, collect lower column liquid, be concentrated into minimum volume after the filtration; With pack into the semi-permeable membranes dialysis tubing dialysis 48 hours of molecular weight cut-off 7000 of liquid concentrator, lyophilize promptly gets the honey raisin tree polysaccharide, and gained honey raisin tree purity of polysaccharide reaches about 70%.
Claims (6)
1. the separation purification method of honey raisin tree polysaccharide is characterized in that comprising the following steps:
(1) polysaccharide extraction liquid preparation: honey raisin tree is pulverized, heating flooding 2-3 time, lixiviate 24-48 hour, filter, merge vat liquor, concentrated honey raisin tree medicinal extract;
(2) alcohol precipitation: the edible ethanol stirring that honey raisin tree medicinal extract is added 3-5 times of 80-90% staticly settled after 5-15 minute, and supernatant is removed in centrifuging, the collecting precipitation thing;
(3) removal of impurities, decolouring: centrifugal sediment is used the hot water stirring and dissolving, last DEAE cellulose column, 0-0.5mol/LNaCl solution gradient wash-out is gone up the ion exchange resin column adsorption bleaching again, and lower column liquid filters the back with the semi-permeable membranes dialysis, is drying to obtain finished product.
2. according to the said separation purification method of claim 1, it is characterized in that the rotating speed of whizzer in the said step (2) is 5000-10000rpm.
3. according to the said separation purification method of claim 1, it is characterized in that cellulose column adopts DEAE-23, DEAE-52 in the said step (3).
4. according to the said separation purification method of claim 1, it is characterized in that optional phenylethylene resin series of said step (3) intermediate ion exchange resin and acrylic resin or both hybrid resins.Phenylethylene resin series can adopt Asmit 265P, DuoliteA171P resin anion(R.A); Acrylic resin can adopt Asmit295, DoliteA172, IRA958S, A860 resin anion(R.A).
5. according to the said separation purification method of claim 1, it is characterized in that semi-permeable membranes is the semi-permeable membranes dialysis tubing of molecular weight cut-off 4000-10000 in the said step (3), dialysed 24-48 hour.
6. according to the said separation purification method of claim 1, it is characterized in that drying means in the said step (3) can adopt a kind of in lyophilize or the spraying drying.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103694371A (en) * | 2014-01-10 | 2014-04-02 | 吉首大学 | Preparation method of honey raisin tree polysaccharide |
| CN107136362A (en) * | 2017-06-13 | 2017-09-08 | 旬阳县太极缘生物科技有限公司 | A kind of honey raisin tree deep process and its honey raisin tree meat, honey raisin tree juice and honey raisin tree slag |
| CN107223824A (en) * | 2017-06-13 | 2017-10-03 | 旬阳县太极缘生物科技有限公司 | A kind of natural selenium-rich honey raisin tree hypoglycemic drink and preparation method thereof |
| CN107858388A (en) * | 2017-10-17 | 2018-03-30 | 陕西科技大学 | A kind of method that sheep breast Antihypertensive Peptides are prepared using probiotic lactobacilli protease |
| CN107853394A (en) * | 2017-10-17 | 2018-03-30 | 陕西科技大学 | A kind of method that lactobacillus bulgaricus protein in cell wall enzyme prepares sheep breast active beverage |
| CN107853396A (en) * | 2017-10-17 | 2018-03-30 | 陕西科技大学 | A kind of method that Lactobacillus rhamnosus protease prepares sheep breast active beverage |
| CN108892732A (en) * | 2018-05-08 | 2018-11-27 | 塔里木大学 | The preparation method and applications of Jinchang jujube polysaccharide with immunoloregulation function |
| CN115975064A (en) * | 2023-01-09 | 2023-04-18 | 天津科技大学 | Hovenia dulcis Thunb polysaccharide with uric acid reducing effect, and preparation method and application thereof |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103694371A (en) * | 2014-01-10 | 2014-04-02 | 吉首大学 | Preparation method of honey raisin tree polysaccharide |
| CN107136362A (en) * | 2017-06-13 | 2017-09-08 | 旬阳县太极缘生物科技有限公司 | A kind of honey raisin tree deep process and its honey raisin tree meat, honey raisin tree juice and honey raisin tree slag |
| CN107223824A (en) * | 2017-06-13 | 2017-10-03 | 旬阳县太极缘生物科技有限公司 | A kind of natural selenium-rich honey raisin tree hypoglycemic drink and preparation method thereof |
| CN107858388A (en) * | 2017-10-17 | 2018-03-30 | 陕西科技大学 | A kind of method that sheep breast Antihypertensive Peptides are prepared using probiotic lactobacilli protease |
| CN107853394A (en) * | 2017-10-17 | 2018-03-30 | 陕西科技大学 | A kind of method that lactobacillus bulgaricus protein in cell wall enzyme prepares sheep breast active beverage |
| CN107853396A (en) * | 2017-10-17 | 2018-03-30 | 陕西科技大学 | A kind of method that Lactobacillus rhamnosus protease prepares sheep breast active beverage |
| CN107853396B (en) * | 2017-10-17 | 2020-12-01 | 陕西科技大学 | Method for preparing goat milk active beverage by using lactobacillus rhamnosus protease |
| CN108892732A (en) * | 2018-05-08 | 2018-11-27 | 塔里木大学 | The preparation method and applications of Jinchang jujube polysaccharide with immunoloregulation function |
| CN108892732B (en) * | 2018-05-08 | 2023-03-31 | 塔里木大学 | Preparation method and application of Jinchang jujube polysaccharide with immunoregulation function |
| CN115975064A (en) * | 2023-01-09 | 2023-04-18 | 天津科技大学 | Hovenia dulcis Thunb polysaccharide with uric acid reducing effect, and preparation method and application thereof |
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Application publication date: 20120530 |