CN105061631A - Extraction method and application of taraxacum polysaccharide - Google Patents
Extraction method and application of taraxacum polysaccharide Download PDFInfo
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Abstract
The invention discloses an extraction method and application of taraxacum polysaccharide. The extraction method is simple, the extraction rate is 8.58%, and the purity is 86.3%. Cyclohexanone is utilized to precipitate the proteins so as to remove the proteins in the extracting solution, so the removal rate is high, thereby obviously enhancing the purity of the polysaccharide extracting solution and having the advantages of low consumption and cost saving. The taraxacum is pulverized and screened to enhance the pulverization degree, so the cell crushing degree is high, and the extraction rate is enhanced. Hot water is used for reflux extraction instead of ethanol, thereby reducing the fat content in the extracting solution, enhancing the polysaccharide purity and lowering the cost. A DEAE-52 (diethylaminoethanol-52) cellulose chromatographic column and a SephadexG-100 gel chromatographic column are utilized to separate and purify the crude polysaccharide, thereby reducing the waste of polysaccharide in the purification process and enhancing the extraction rate. The taraxacum polysaccharide can be used as an oxidation-resistant active component for preparing the health food or medicine, has the advantages of obvious antioxidation effect, high safety and no side effect, and has great market value and wide application prospects.
Description
Technical field
The present invention relates to medical art, particularly a kind of extracting method of Taraxacum Polysaccharides and application thereof.
Background technology
Taraxacum, property is flat, sweet, the micro-hardship of taste.Can be clearing heat and detoxicating, dispersing swelling and dissipating binds.Have significant galgctogogue action, treatment mazoitis is very effective.No matter decoct juice oral, still smash mud external application, all have effect.In addition, taraxacum also has diuresis, laxative, removing jaundice subcutaneous ulcer, cholagogic, aid digestion, and increase appetite, treatment gastric and duodenal ulcer, also can prevent and treat various tumour.It has abundant vitamin A, C and mineral substance, to the effect that maldigestion, constipation are all improved.Leaf is also improved eczema in addition, clean blood effect of dermatitis of releiving, joint discomfort, and root then has antiinflammation, can treat gallbladdergallstonecholetithiasis, rheumatism, and flower decocts into concoction can remove freckle, can say it is very useful a kind of incense drug grass.Containing multiple health-nutrition compositions such as taraxol, taraxacin, choline, organic acid, synanthrin in taraxacum plant materials, there are the effects such as diuresis, laxative, removing jaundice subcutaneous ulcer, cholagogic.Taraxacum, simultaneously containing protein, fat, carbohydrate, trace element and VITAMIN etc., has abundant nutritive value, raw-eaten, stir-fry and eat, cook soup, is the plant of dietotherapeutic.Dandelion plant is rich in sterol, palmitinic acid and polysaccharide, and polysaccharide is important informational molecule, have antitumor, antiviral, anti-ageing, radioprotective, anti-mutation, hypoglycemic, strengthen many-sided effects such as immunity.
Summary of the invention
In order to make up the defect of prior art, the object of this invention is to provide a kind of extracting method and application thereof of Taraxacum Polysaccharides.
An extracting method for Taraxacum Polysaccharides, its concrete steps are:
(1) herb taraxacum is cleaned, dry, 45 DEG C of oven dry, pulverize, cross 60 ~ 100 mesh sieves;
(2) get dry powder 100g, by solid-liquid ratio 1:8 ~ 12, add deionized water, 95 ~ 100 DEG C of water-baths, reflux 4 ~ 5 times, each 1 hour, filter, obtain extracting solution;
(3) in extracting solution, slowly pimelinketone is added, stirring while adding, is 8 ~ 12% of extracting liquid volume, 1000 revs/min to add-on, magnetic agitation 20 ~ 30 minutes, leave standstill 140 ~ 160 minutes, 10000 revs/min, centrifugal 10 minutes, get supernatant, 40 ~ 45 DEG C of rotary evaporation in vacuo are to 1/3 of original volume, and collecting ring hexanone, reuses;
(4) to 3) add dehydrated alcohol in the supernatant that obtains, stir while adding, account for 85% of cumulative volume to ethanol contend, be placed in 4 DEG C of refrigerators, hold over night, 8000 revs/min, centrifugal 15 minutes, get precipitation;
(5) to 4) add the deionized water of 3 ~ 4 times amount in the precipitation that obtains, vacuum rotating is concentrated into without ethanol, obtains Crude polysaccharides solution;
(6) poured in 7000 daltonian dialysis tubings by Crude polysaccharides solution, dialysis tubing is placed in the beaker filling deionized water, dialyse 36 ~ 48 hours for 4 DEG C, the deionized water in replacing beaker 3 ~ 4 times, dialysis tubing can be reused;
(7) the Crude polysaccharides solution after dialysis is splined on DEAE-52 Mierocrystalline cellulose chromatography column with the speed of 0.5ml/min, use NaCl solution and the deionized water wash-out of 1mol/L successively, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve, collect each position polysaccharide elutriant according to elution curve, DPPH method measures the anti-oxidant activity of each position polysaccharide, and rotary evaporation concentrates final vacuum lyophilize;
(8) each position polysaccharide deionized water through DEAE-52 Mierocrystalline cellulose chromatography column initial gross separation purifying is mixed with the solution of 5mg/ml, draw 2ml and be splined on SephadexG-100 gel chromatography column, wash-out is carried out with deionized water, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve, merge sugared peak elutriant, rotary evaporation concentrates final vacuum lyophilize, obtains purifying Taraxacum Polysaccharides.
Preferably, described step 1) crosses 80 mesh sieves after being pulverized by taraxacum.
Preferably, described step 2) the solid-liquid ratio 1:12 of distilled water backflow, 100 DEG C of water-baths, reflux 5 times.
The application of a kind of Taraxacum Polysaccharides of the present invention is for the preparation of protective foods or medicine using described Taraxacum Polysaccharides as oxidation-resistant active ingredient.
Advantage of the present invention is:
1, the extracting method of a kind of Taraxacum Polysaccharides provided by the invention and application thereof, extracting method is simple, extraction yield is 8.58%, purity is 86.3%, uses pimelinketone protein precipitation, removes the protein in extracting solution, clearance is high, can significantly improve the purity of polysaccharide extraction liquid, and usage quantity is few, cost-saving;
2, by taraxacum grinding and sieving, improve degree of grinding, the degree of crushing of cell is high, improves extraction yield; Replace ethanol to carry out refluxing extraction with hot water, reduce the lipid content in extracting solution, improve the purity of polysaccharide, reduce subsequent purification process, reduce costs;
3, DEAE-52 Mierocrystalline cellulose chromatography column and SephadexG-100 gel chromatography column is utilized to carry out separation and purification to Crude polysaccharides, with an organic solvent, do not reduce the waste to polysaccharide in purge process, improve extraction yield, be conducive to being in charge of collection, measure sugared content and the anti-oxidant activity of different sites;
4, Taraxacum Polysaccharides can be used as oxidation-resistant active ingredient for the preparation of protective foods or medicine, and antioxygenation is obvious, and safety, has no side effect, and has very large marketable value and wide application prospect.
Accompanying drawing explanation
Fig. 1 is DEAE-52 Mierocrystalline cellulose chromatography column elution curve of the present invention;
Fig. 2 is the anti-oxidant activity figure that SephadexG-100 gel chromatography column of the present invention respectively collects peak;
Fig. 3 is SephadexG-100 gel chromatography column elution curve of the present invention;
Fig. 4 is SephadexG-100 gel chromatography column elution curve of the present invention;
Fig. 5 is SephadexG-100 gel chromatography column elution curve of the present invention;
Fig. 6 is SephadexG-100 gel chromatography column elution curve of the present invention;
Fig. 7 is SephadexG-100 gel chromatography column elution curve of the present invention;
Fig. 8 is SephadexG-100 gel chromatography column elution curve of the present invention.
Embodiment
With specific embodiment, the present invention is described below.
Embodiment 1
An extracting method for Taraxacum Polysaccharides, its concrete steps are:
(1) herb taraxacum is cleaned, dry, 45 DEG C of oven dry, pulverize, cross 80 mesh sieves;
(2) get dry powder 100g, by solid-liquid ratio 1:12, add deionized water, 100 DEG C of water-baths, reflux 5 times, each 1 hour, filter, obtain extracting solution;
(3) in extracting solution, slowly pimelinketone is added, stirring while adding, is 12% of extracting liquid volume, 1000 revs/min to add-on, magnetic agitation 20 minutes, leave standstill 160 minutes, 10000 revs/min, centrifugal 10 minutes, get supernatant, 45 DEG C of rotary evaporation in vacuo are to 1/3 of original volume, and collecting ring hexanone, reuses;
(4) to 3) add dehydrated alcohol in the supernatant that obtains, stir while adding, account for 85% of cumulative volume to ethanol contend, be placed in 4 DEG C of refrigerators, hold over night, 8000 revs/min, centrifugal 15 minutes, get precipitation;
(5) to 4) add the deionized water of 3 times amount in the precipitation that obtains, vacuum rotating is concentrated into without ethanol, obtains Crude polysaccharides solution;
(6) poured in 7000 daltonian dialysis tubings by Crude polysaccharides solution, dialysis tubing is placed in the beaker filling deionized water, dialyse 48 hours for 4 DEG C, the deionized water in replacing beaker 4 times, dialysis tubing can be reused;
(7) the Crude polysaccharides solution after dialysis is splined on DEAE-52 Mierocrystalline cellulose chromatography column with the speed of 0.5ml/min, use NaCl solution and the deionized water wash-out of 1mol/L successively, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve (see figure 1), collect each position polysaccharide elutriant according to elution curve, DPPH method measures the anti-oxidant activity (see figure 2) of each position polysaccharide, and rotary evaporation concentrates final vacuum lyophilize;
(8) each position polysaccharide deionized water through DEAE-52 Mierocrystalline cellulose chromatography column initial gross separation purifying is mixed with the solution of 5mg/ml, draw 2ml and be splined on SephadexG-100 gel chromatography column, carry out wash-out with deionized water, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content (see Fig. 3 ~ 8), draw elution curve, merge sugared peak elutriant, rotary evaporation concentrates final vacuum lyophilize, obtains purifying Taraxacum Polysaccharides, extraction yield is 8.58% of herb quality, and purity is 86.3%.
The application of a kind of Taraxacum Polysaccharides of the present invention is for the preparation of protective foods or medicine using described Taraxacum Polysaccharides as oxidation-resistant active ingredient.
Embodiment 2
An extracting method for Taraxacum Polysaccharides, its concrete steps are:
(1) herb taraxacum is cleaned, dry, 45 DEG C of oven dry, pulverize, cross 60 mesh sieves;
(2) get dry powder 100g, by solid-liquid ratio 1:8, add deionized water, 100 DEG C of water-baths, reflux 5 times, each 1 hour, filter, obtain extracting solution;
(3) in extracting solution, slowly pimelinketone is added, stirring while adding, is 10% of extracting liquid volume, 1000 revs/min to add-on, magnetic agitation 30 minutes, leave standstill 140 minutes, 10000 revs/min, centrifugal 10 minutes, get supernatant, 43 DEG C of rotary evaporation in vacuo are to 1/3 of original volume, and collecting ring hexanone, reuses;
(4) to 3) add dehydrated alcohol in the supernatant that obtains, stir while adding, account for 85% of cumulative volume to ethanol contend, be placed in 4 DEG C of refrigerators, hold over night, 8000 revs/min, centrifugal 15 minutes, get precipitation;
(5) to 4) add the deionized water of 3 times amount in the precipitation that obtains, vacuum rotating is concentrated into without ethanol, obtains Crude polysaccharides solution;
(6) poured in 7000 daltonian dialysis tubings by Crude polysaccharides solution, dialysis tubing is placed in the beaker filling deionized water, dialyse 40 hours for 4 DEG C, the deionized water in replacing beaker 4 times, dialysis tubing can be reused;
(7) the Crude polysaccharides solution after dialysis is splined on DEAE-52 Mierocrystalline cellulose chromatography column with the speed of 0.5ml/min, use NaCl solution and the deionized water wash-out of 1mol/L successively, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve, collect each position polysaccharide elutriant according to elution curve, DPPH method measures the anti-oxidant activity of each position polysaccharide, and rotary evaporation concentrates final vacuum lyophilize;
(8) each position polysaccharide deionized water through DEAE-52 Mierocrystalline cellulose chromatography column initial gross separation purifying is mixed with the solution of 5mg/ml, draw 2ml and be splined on SephadexG-100 gel chromatography column, carry out wash-out with deionized water, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve, merge sugared peak elutriant, rotary evaporation concentrates final vacuum lyophilize, obtains purifying Taraxacum Polysaccharides, extraction yield is 7.35% of herb quality, and purity is 84.2%.
The application of a kind of Taraxacum Polysaccharides of the present invention is for the preparation of protective foods or medicine using described Taraxacum Polysaccharides as oxidation-resistant active ingredient.
Embodiment 3
An extracting method for Taraxacum Polysaccharides, its concrete steps are:
(1) herb taraxacum is cleaned, dry, 45 DEG C of oven dry, pulverize, cross 100 mesh sieves;
(2) get dry powder 100g, by solid-liquid ratio 1:10, add deionized water, 95 DEG C of water-baths, reflux 4 times, each 1 hour, filter, obtain extracting solution;
(3) in extracting solution, slowly pimelinketone is added, stirring while adding, is 8% of extracting liquid volume, 1000 revs/min to add-on, magnetic agitation 20 minutes, leave standstill 140 minutes, 10000 revs/min, centrifugal 10 minutes, get supernatant, 40 DEG C of rotary evaporation in vacuo are to 1/3 of original volume, and collecting ring hexanone, reuses;
(4) to 3) add dehydrated alcohol in the supernatant that obtains, stir while adding, account for 85% of cumulative volume to ethanol contend, be placed in 4 DEG C of refrigerators, hold over night, 8000 revs/min, centrifugal 15 minutes, get precipitation;
(5) to 4) add the deionized water of 3 ~ 4 times amount in the precipitation that obtains, vacuum rotating is concentrated into without ethanol, obtains Crude polysaccharides solution;
(6) poured in 7000 daltonian dialysis tubings by Crude polysaccharides solution, dialysis tubing is placed in the beaker filling deionized water, dialyse 48 hours for 4 DEG C, the deionized water in replacing beaker 3 times, dialysis tubing can be reused;
(7) the Crude polysaccharides solution after dialysis is splined on DEAE-52 Mierocrystalline cellulose chromatography column with the speed of 0.5ml/min, use NaCl solution and the deionized water wash-out of 1mol/L successively, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve, collect each position polysaccharide elutriant according to elution curve, DPPH method measures the anti-oxidant activity of each position polysaccharide, and rotary evaporation concentrates final vacuum lyophilize;
(8) each position polysaccharide deionized water through DEAE-52 Mierocrystalline cellulose chromatography column initial gross separation purifying is mixed with the solution of 5mg/ml, draw 2ml and be splined on SephadexG-100 gel chromatography column, carry out wash-out with deionized water, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve, merge sugared peak elutriant, rotary evaporation concentrates final vacuum lyophilize, obtains purifying Taraxacum Polysaccharides, extraction yield is 8.85% of herb quality, and purity is 81.5%.
The application of a kind of Taraxacum Polysaccharides of the present invention is for the preparation of protective foods or medicine using described Taraxacum Polysaccharides as oxidation-resistant active ingredient.
Claims (4)
1. an extracting method for Taraxacum Polysaccharides, is characterized in that, its concrete steps are:
(1) herb taraxacum is cleaned, dry, 45 DEG C of oven dry, pulverize, cross 60 ~ 100 mesh sieves;
(2) get taraxacum dry powder 100g, by solid-liquid ratio 1:8 ~ 12, add deionized water, 95 ~ 100 DEG C of water-baths, reflux 4 ~ 5 times, each 1 hour, filter, obtain extracting solution;
(3) in extracting solution, slowly pimelinketone is added, stirring while adding, is 8 ~ 12% of extracting liquid volume, 1000 revs/min to add-on, magnetic agitation 20 ~ 30 minutes, leave standstill 140 ~ 160 minutes, 10000 revs/min, centrifugal 10 minutes, get supernatant, 40 ~ 45 DEG C of rotary evaporation in vacuo are to 1/3 of original volume, and collecting ring hexanone, reuses;
(4) to 3) add dehydrated alcohol in the supernatant that obtains, stir while adding, account for 85% of cumulative volume to ethanol contend, be placed in 4 DEG C of refrigerators, hold over night, 8000 revs/min, centrifugal 15 minutes, get precipitation;
(5) to 4) add the deionized water of 3 ~ 4 times amount in the precipitation that obtains, vacuum rotating is concentrated into without ethanol, obtains Crude polysaccharides solution;
(6) poured in 7000 daltonian dialysis tubings by Crude polysaccharides solution, dialysis tubing is placed in the beaker filling deionized water, dialyse 36 ~ 48 hours for 4 DEG C, the deionized water in replacing beaker 3 ~ 4 times, dialysis tubing can be reused;
(7) the Crude polysaccharides solution after dialysis is splined on DEAE-52 Mierocrystalline cellulose chromatography column with the speed of 0.5ml/min, use NaCl solution and the deionized water wash-out of 1mol/L successively, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve, collect each position polysaccharide elutriant according to elution curve, DPPH method measures the anti-oxidant activity of each position polysaccharide, and rotary evaporation concentrates final vacuum lyophilize;
(8) each position polysaccharide deionized water through DEAE-52 Mierocrystalline cellulose chromatography column initial gross separation purifying is mixed with the solution of 5mg/ml, draw 2ml and be splined on SephadexG-100 gel chromatography column, wash-out is carried out with deionized water, be in charge of collection, collect liquid phend-sulphuric acid and detect polysaccharide content, draw elution curve, merge sugared peak elutriant, rotary evaporation concentrates final vacuum lyophilize, obtains purifying Taraxacum Polysaccharides.
2. the extracting method of a kind of Taraxacum Polysaccharides as claimed in claim 1, is characterized in that, preferably, described step 1) crosses 80 mesh sieves after being pulverized by taraxacum.
3. the extracting method of a kind of Taraxacum Polysaccharides as claimed in claim 1, is characterized in that, preferably, described step 2) the solid-liquid ratio 1:12 of distilled water backflow, 100 DEG C of water-baths, reflux 5 times.
4. the application of a kind of Taraxacum Polysaccharides as claimed in claim 1, is characterized in that, using described Taraxacum Polysaccharides as oxidation-resistant active ingredient for the preparation of protective foods or medicine.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110128559A (en) * | 2019-04-29 | 2019-08-16 | 华南农业大学 | A kind of preparation method and applications with immunoregulatory fire ginseng fruit pericarp polysaccharide |
| CN110437344A (en) * | 2019-09-17 | 2019-11-12 | 广西民族大学 | A kind of dendrobium candidum refined polysaccharide and preparation method thereof |
| CN113425757A (en) * | 2021-06-08 | 2021-09-24 | 中国中医科学院中医基础理论研究所 | Dandelion extract and preparation method and application thereof |
| CN113621085A (en) * | 2021-07-08 | 2021-11-09 | 新疆农业科学院农业机械化研究所 | Preparation method of dandelion root polysaccharide sulfation modification product and application of dandelion root polysaccharide sulfation modification product |
| CN113698507A (en) * | 2021-10-18 | 2021-11-26 | 东北农业大学 | Preparation method and application of carboxymethylated dandelion polysaccharide |
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| US20110275142A1 (en) * | 2010-05-06 | 2011-11-10 | James Wade | Dandelion processes, compositions and products |
| CN102653568A (en) * | 2012-05-28 | 2012-09-05 | 上海中医药大学 | Herba taraxaci homogeneous polysaccharide and preparation method and application thereof |
| CN102659958A (en) * | 2012-05-28 | 2012-09-12 | 上海中医药大学 | Dandelion polysaccharide extract and preparation method and application thereof |
| CN103275175A (en) * | 2013-05-19 | 2013-09-04 | 陕西理工学院 | Preparation method of dandelion small molecule glycoprotein |
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2015
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20110275142A1 (en) * | 2010-05-06 | 2011-11-10 | James Wade | Dandelion processes, compositions and products |
| CN102653568A (en) * | 2012-05-28 | 2012-09-05 | 上海中医药大学 | Herba taraxaci homogeneous polysaccharide and preparation method and application thereof |
| CN102659958A (en) * | 2012-05-28 | 2012-09-12 | 上海中医药大学 | Dandelion polysaccharide extract and preparation method and application thereof |
| CN103275175A (en) * | 2013-05-19 | 2013-09-04 | 陕西理工学院 | Preparation method of dandelion small molecule glycoprotein |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110128559A (en) * | 2019-04-29 | 2019-08-16 | 华南农业大学 | A kind of preparation method and applications with immunoregulatory fire ginseng fruit pericarp polysaccharide |
| CN110128559B (en) * | 2019-04-29 | 2021-02-19 | 华南农业大学 | Preparation method and application of polysaccharide of fruit peel of fire ginseng with immunoregulation function |
| CN110437344A (en) * | 2019-09-17 | 2019-11-12 | 广西民族大学 | A kind of dendrobium candidum refined polysaccharide and preparation method thereof |
| CN113425757A (en) * | 2021-06-08 | 2021-09-24 | 中国中医科学院中医基础理论研究所 | Dandelion extract and preparation method and application thereof |
| CN113621085A (en) * | 2021-07-08 | 2021-11-09 | 新疆农业科学院农业机械化研究所 | Preparation method of dandelion root polysaccharide sulfation modification product and application of dandelion root polysaccharide sulfation modification product |
| CN113698507A (en) * | 2021-10-18 | 2021-11-26 | 东北农业大学 | Preparation method and application of carboxymethylated dandelion polysaccharide |
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