CN101580552A - Preparation method of urtica angustifolia polysaccharide and application thereof - Google Patents
Preparation method of urtica angustifolia polysaccharide and application thereof Download PDFInfo
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- CN101580552A CN101580552A CNA2009100671772A CN200910067177A CN101580552A CN 101580552 A CN101580552 A CN 101580552A CN A2009100671772 A CNA2009100671772 A CN A2009100671772A CN 200910067177 A CN200910067177 A CN 200910067177A CN 101580552 A CN101580552 A CN 101580552A
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Abstract
The invention relates to a preparation method of urtica angustifolia polysaccharide and an application thereof, belonging to the fields of food and medicine. The method comprises the following steps of: 1, taking urtica angustifolia, drying the urtica angustifolia in the shade, pulverizing the urtica angustifolia, adding water of 8 to 10 times in quantity, heating the mixture to 100 DEG C, and leaching the mixture for 2 to 3 times lasting for 2 to 3 hours every time; 2, filtering the mixture, merging filter liquor, carrying out depressurization concentration and concentrating the mixture to one twelve of the volume of the original liquor; 3, carrying out centrifugation for 15 minutes by 3500 rev/min, taking supernatant fluid and carrying out alcohol precipitation on 95% of ethanol; and 4, carrying out deproteinization for 3 times by a Sevage method, carrying out centrifugation for 10 minutes by 3500 rev/min, concentrating the mixture to volatilize a solvent and carrying out freeze drying. The invention also relates to the application of the urtica angustifolia polysaccharide in the preparation of medicine with an anti-fatigue function. The medicine has obvious anti-fatigue effect, safety and no toxicity.
Description
Technical field
The invention belongs to food, medical technical field, refer in particular to the preparation of urtica angustifolia polysaccharide and the purposes of antifatigue thereof.
Background technology
Nettle is Angiospermae, Urticaceae, Urtica (Urtica.L.), annual or per nnial herb, and it is hot, bitter, warm, puckery, slightly poisonous to distinguish the flavor of.Vitality is extremely strong, and natural resource are abundant, especially at China's northeast resource horn of plenty more.Urtica angustifolia (Urtica angustifolia) also is called and bites fiber crops, per nnial herb.Stem is high 40～150 centimetres, and four prismatics have stinging hair, branch or branch not.Leaf is to life, lanceolar or narrow avette, and long 4～10 centimetres, wide 1.2～2.8 centimetres, tip is point gradually, the base portion circle, there is sharp sawtooth at the edge, dredges above and gives birth to undercoat, has to dredge on along the pulse below and gives birth to undercoat; Stipule is mitogenetic, bar shaped.Dioecy, inflorescence reach 4 centimetres, multi-branched; Male flower tapel 4, stamen 4; Female flower is little than male flower, and tapel 4 increases column cap paintbrush head in the fruit phase.Follicarpium is avette, and is flat, is about 1 millimeter, smooth.The regional northeastward diseases such as rheumatic arthritis, urticaria, snakebite, diabetes that are usually used in treating among the people.Have and dispel rheumatism, promoting blood circulation and stopping pain, flat liver arresting convulsion, effects such as long-pending defaecation, detoxifcation disappear.Multiple function such as polysaccharide has immunomodulatory, antitumor, anti-inflammatory, antiviral, anti-oxidant, radioprotective, hypoglycemic, reducing blood-fat, protect the liver.
Summary of the invention
The invention provides preparation of a kind of urtica angustifolia polysaccharide and uses thereof, urtica angustifolia polysaccharide has the function of antifatigue.
The extracting method of urtica angustifolia polysaccharide polysaccharide of the present invention is:
One, get urtica angustifolia, dry in the shade, pulverize, add 8～10 times of amounts water, be heated to 100 ℃, lixiviate 2～3 times, each 2～3 hours;
Two, filter, merging filtrate, is concentrated into stoste volume 1/12 at concentrating under reduced pressure,
Three, with 3500 rev/mins, centrifugal 15 minutes, get supernatant liquor, 95% ethanol alcohol precipitation;
Four, Sevage method deproteinated is 3 times, with 3500 rev/mins, centrifugal 10 minutes, concentrates and flings to solvent, lyophilize.
Urtica angustifolia polysaccharide of the present invention has application in the medicine of anti-fatigue effect in preparation.
The present invention also provides the various new drugs with urtica angustifolia polysaccharide and pharmaceutically acceptable carrier or vehicle preparation.These new drugs comprise oral Preparation, Sublingual tablet, suppository, effervescent tablet, injection etc.Oral Preparation comprises oral solid formulation, oral liquid.Oral solid formulation can be tablet, capsule, granule, pill, lozenge etc.In these preparations, except that comprising total effective parts of the present invention, can also choose wantonly and contain pharmaceutically acceptable weighting agent, glidant, lubricant, tinting material, correctives etc.These preparations can also be prepared into slowly-releasing or controlled release preparation as required.Oral liquid can be solution, suspension, emulsion etc.In these preparations, except that comprising total effective parts of the present invention, can also choose wantonly and contain pharmaceutically acceptable solvent, solubility promoter, solubilizing agent, suspending agent, emulsifying agent, tensio-active agent, correctives etc.
The new drug that with the urtica angustifolia polysaccharide is raw material makes provided by the invention not only comprises single preparations of ephedrine, also is included in to add the medicine of urtica angustifolia polysaccharide as activeconstituents in the compound preparation.
The invention has the beneficial effects as follows: the effect of antifatigue is obvious, and urtica angustifolia polysaccharide people consumption is 0.1～1.5 gram/sky, has the function of antifatigue, and the safety non-toxic effect.
Embodiment
Following pharmacodynamic experiment by urtica angustifolia polysaccharide further specifies the present invention.
1, swimming with a load attached to the body test
Inspection principle, the raising of exercise tolerance are that anti-fatigue ability is strengthened the most direct performance, and the length of swimming time can be reacted the degree of animal movement fatigue.
Instrument and equipment:
The swimming case (the about 50cm * 50cm of size * 40cm);
The AL104 electronic balance, plum Teller-Tuo benefit Instr Ltd.;
Aluminium wire;
Laboratory animal: cleaning level Kunming mouse, body weight 18～22g.
The dosage grouping:
Three dosage groups and a blank group are established in experiment, and basic, normal, high dosage group is subjected to the examination amount to be respectively 100mg/Kg, 200mg/Kg, 300mg/Kg, and concentration is respectively 4g/ml, 8g/ml, 12g/ml.
Experimental procedure:
After last gives given the test agent 30min, put mouse and in the swimming case, swim.The depth of water is no less than 30cm, 25 ℃ ± 1.0 ℃ of water temperatures, the load aluminium wire of 5% body weight of mouse root of the tail portion.The record mouse is from the extremely dead time of swimming beginning, as the mouse swimming time.
Experimental result and analysis:
Table 1 polysaccharide is to the influence of mouse swimming with a load attached to the body time (X ± SD)
*; There were significant differences (P＜0.05) with the control group ratio
By table 1 as seen, compare with control group, the low dose group swimming time has improved 85.4%, and middle dosage group swimming time has improved 114.6%, and the high dose group swimming time has improved 151.2% (P＜0.05).
2, the mensuration of serum urea nitrogen
Test method: after last is tried thing 30min to mouse, be not swimming with a load attached to the body 90min in 30 ℃ the water, dial eyeball behind the rest 60min and get blood in temperature.The centrifugal 15min of 2000r/min behind 4 ℃ of static 3h takes out serum, measures the serum urea nitrogen content with automatic clinical chemistry analyzer.
Mouse blood urea nitrogen experimental result and analysis in table 2:
Table 2 polysaccharide is to the influence of mice serum blood urea nitrogen (X ± SD)
With control group than * P＜0.05; Material P＜0.01
By table 2 as seen, compare with control group, the serum urea nitrogen behind the low dose group mouse movement has reduced by 12.8% (P＜0.05); Middle dosage group has reduced by 13.6% (P＜0.05); High dose group has reduced by 19.8% (P＜0.01).
3, liver starch is measured: anthrone method.
Detect principle: anthrone can react with free sugar or polysaccharide, and reaction back solution is blue-greenish colour, in the 620nm place maximum absorption is arranged.Measure its optical density(OD), can determine content of glycogen.
Instrument and reagent
723 type spectrophotometers, glad science equipment development company of Liaoning section,
The Z200A whizzer, German Harmer (Hermle),
Fsh-2 is adjustable high speed electric homogenizer, permanent laboratory apparatus company limited in the Wuxi, vibrator moves liquid pump, and boiling water bath is irritated the stomach syringe needle, instruments, glass funnel, sample injector, 2mL, 5mL, 10mL valinche, 20mL band plug scale test tube, 10mL band plug centrifuge tube.
Reagent: 5% Tricholroacetic Acid (joining) with distilled water (TCA), glucose reference liquid, the vitriol oil (AR), anthrone reagent.
Anthrone reagent: contain 0.05% anthrone in the solution, 1% thiocarbamide, H with 72%
2SO
4Preparation.Compound method is as follows: 1. 72%H
2SO
4Preparation: add 280mL distilled water in the beaker, add highly purified vitriol oil 720mL (proportion 1.84) again.2. anthrone reagent is joined method: work as H
2SO
4Put into the pure anthrone of 500mg when temperature is reduced to 80-90 ℃, the highly purified thiocarbamide of 10g suitably shakes the beaker mixing.Deposit in after the cooling in the refrigerator, can preserve for two weeks.
Experimental technique
Laboratory animal: cleaning level Kunming mouse, body weight 18-22g.
Dosage design and grouping,
Three dosage groups and a blank group are established in experiment, and basic, normal, high dosage group is subjected to the examination amount to be respectively 100mg/Kg, 200mg/Kg, 300mg/Kg, and concentration is respectively 4g/ml, 8g/ml, 12g/ml.
Experimental procedure:
30min put to death animal after last was given sample, got liver and blotted with filter paper after the physiological saline rinsing, accurately took by weighing liver 100mg, add 8mL TCA, every pipe homogenate 1min pours homogenate into centrifuge tube, with the centrifugal 15min of 3000 commentaries on classics/min, supernatant liquor is transferred to another in vitro.Get the 1mL supernatant liquor and put into the 10mL centrifuge tube, every pipe adds 95% ethanol 4mL, does not fully leave the interface between mixing to two kind of liquid.With clean stopper beyond the Great Wall, spend the night (also can select for use test tube is placed on 37-40 ℃ of water-bath 3h) placed in setting under the room temperature.Precipitation fully after, with test tube in the centrifugal 15min of 3000 commentaries on classics/min.Carefully outwell supernatant liquor and test tube is stood upside down and place 10min 2mL dissolved in distilled water glycogen, the glycogen with tube wall when adding water washes.As dissolving immediately of the glycogen of managing the end, the vibration pipe is up to dissolving fully.
Make reagent blank and standard pipe:
Reagent blank: inhale 2mL distilled water to clean centrifuge tube;
Standard pipe: inhale 0.5mL glucose reference liquid, contain 100mg/dL glucose and 1.5mL distilled water is put into same pipe.
Firmly add each pipe with the 10mL anthrone reagent this moment, and liquid stream (anthrone reagent) directly enters pipe central authorities, guarantees that thorough mixing is good.When from pipe, injecting anthrone reagent, pipe is placed under the cold water faucet has a shower.After all pipes all reach the cold water temperature, it is dipped in boiling water bath (the water-bath degree of depth is a little more than the pipe liquid level) 15min, move on to cooling bath then, cool to room temperature.Liquid in pipe is moved into colorimetric cylinder, under the 620nm wavelength, measure absorbancy with reagent blank pipe zeroing back.And calculate glycogen content according to typical curve, be converted into hepatic glycogen content (representing) according to the liver weight that is taken by weighing, and carry out statistical study with the mg/g liver.
Glycogen content calculates:
DU: the sample hose absorbancy,
DS: the standard pipe absorbancy,
0.5: be the glucose content in the 0.5mL glucose reference liquid.
0.9: for glucose being converted into the coefficient of glycogen.
Extracting liquid volume: be 8ML;
Hepatic tissue gram number: be 0.1g;
Experimental result and analysis
Table 3 polysaccharide is to the influence of Mouse Liver glycogen (X ± SD)
With control group than * * P＜0.01
By table 3 as seen, compare with control group, the hepatic glycogen content behind the low dose group mouse movement has improved 87.0%; Middle dosage group has improved 469.6% (P＜0.01); High dose group has improved 747.8% (P＜0.01).
4, result and discussion
After per os was irritated 10 days polysaccharide of stomach mouse, compare with control group: low dosage, middle dosage, high dosage had all improved the swimming time of mouse, and high dose group has been compared statistical meaning with the blank group, and P＜0.05; The post exercise mouse is compared with the blank group, and the serum urea nitrogen content all is lower than the blank group, and all has statistical significance, low dose group and blank P＜0.05, middle dosage group and blank P＜0.05, high dose group and blank P＜0.01; Hepatic glycogen content all is higher than the blank group, and middle dosage and high dose group all have statistical significance, and middle dosage group is compared P＜0.01 with the blank group; High dose group is compared P＜0.01 with the blank group.Therefore, the middle and high dosage group polysaccharide effect that mouse had antifatigue.
The preparation of embodiment 1 urtica angustifolia polysaccharide
One, get urtica angustifolia, dry in the shade, pulverize, add 8 times of amounts water, be heated to 100 ℃, lixiviate 3 times, each 3 hours;
Two, filter, merging filtrate, is concentrated into stoste volume 1/12 at concentrating under reduced pressure,
Three, with 3500 rev/mins, centrifugal 15 minutes, get supernatant liquor, 95% ethanol alcohol precipitation;
Four, Sevage method deproteinated is 3 times, with 3500 rev/mins, centrifugal 10 minutes, concentrates and flings to solvent, lyophilize.
Embodiment 2
One, get urtica angustifolia, dry in the shade, pulverize, add 9 times of amounts water, be heated to 100 ℃, lixiviate 3 times, each 2.5 hours;
Two, filter, merging filtrate, is concentrated into stoste volume 1/12 at concentrating under reduced pressure,
Three, with 3500 rev/mins, centrifugal 15 minutes, get supernatant liquor, 95% ethanol alcohol precipitation;
Four, Sevage method deproteinated is 3 times, with 3500 rev/mins, centrifugal 10 minutes, concentrates and flings to solvent, lyophilize.
Embodiment 3
One, get urtica angustifolia, dry in the shade, pulverize, add 8 times of amounts water, be heated to 100 ℃, lixiviate 2 times, each 2 hours;
Two, filter, merging filtrate, is concentrated into stoste volume 1/12 at concentrating under reduced pressure,
Three, with 3500 rev/mins, centrifugal 15 minutes, get supernatant liquor, 95% ethanol alcohol precipitation;
Four, Sevage method deproteinated is 3 times, with 3500 rev/mins, centrifugal 10 minutes, concentrates and flings to solvent, lyophilize.
The preparation of embodiment 4 capsules of the present invention
Urtica angustifolia polysaccharide 50.00g, medical starch 200.00g, the two thorough mixing, encapsulated, make 1000 capsules, every heavy 0.25g contains urtica angustifolia polysaccharide 50mg.
The preparation of embodiment 5 tablets of the present invention
Urtica angustifolia polysaccharide 25.00g, the conventional auxiliary material 225.00g of preparation tablet, compressing tablet is made 1000, and every heavy 0.25g contains urtica angustifolia polysaccharide 25mg.
Claims (2)
1, a kind of preparation method of urtica angustifolia polysaccharide comprises the following steps:
One, get urtica angustifolia, dry in the shade, pulverize, add 8～10 times of amounts water, be heated to 100 ℃, lixiviate 2～3 times, each 2～3 hours;
Two, filter, merging filtrate, is concentrated into stoste volume 1/12 at concentrating under reduced pressure,
Three, with 3500 rev/mins, centrifugal 15 minutes, get supernatant liquor, 95% ethanol alcohol precipitation;
Four, Sevage method deproteinated is 3 times, with 3500 rev/mins, centrifugal 10 minutes, concentrates and flings to solvent, lyophilize.
2, urtica angustifolia polysaccharide has application in the medicine of anti-fatigue effect in preparation.
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| Application Number | Priority Date | Filing Date | Title |
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| CNA2009100671772A CN101580552A (en) | 2009-06-24 | 2009-06-24 | Preparation method of urtica angustifolia polysaccharide and application thereof |
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| CNA2009100671772A CN101580552A (en) | 2009-06-24 | 2009-06-24 | Preparation method of urtica angustifolia polysaccharide and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104522461A (en) * | 2014-12-04 | 2015-04-22 | 黄艳红 | Glossy ganoderma peptide rhizoma polygonati polysaccharide and preparation technology thereof |
| CN106084081A (en) * | 2016-06-16 | 2016-11-09 | 大兴安岭至臻尚品寒带生物技术有限公司 | The preparation method of polysaccharide component and products application in a kind of Daxing'an Mountainrange urtica angustifolia and Betula Phellinus igniarius (L. ex Fr.) Quel. |
| CN112812198A (en) * | 2021-01-13 | 2021-05-18 | 西藏新浩药业有限公司 | Method for extracting polysaccharide from nettle |
| CN116731222A (en) * | 2023-07-13 | 2023-09-12 | 昆明市延安医院 | Nettle rhamnogalacturonan and preparation method and application thereof |
-
2009
- 2009-06-24 CN CNA2009100671772A patent/CN101580552A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104522461A (en) * | 2014-12-04 | 2015-04-22 | 黄艳红 | Glossy ganoderma peptide rhizoma polygonati polysaccharide and preparation technology thereof |
| CN106084081A (en) * | 2016-06-16 | 2016-11-09 | 大兴安岭至臻尚品寒带生物技术有限公司 | The preparation method of polysaccharide component and products application in a kind of Daxing'an Mountainrange urtica angustifolia and Betula Phellinus igniarius (L. ex Fr.) Quel. |
| CN106084081B (en) * | 2016-06-16 | 2018-09-21 | 大兴安岭至臻尚品寒带生物技术有限公司 | The preparation method and products application of polysaccharide component in a kind of Daxing'an Mountainrange urtica angustifolia and birch Phellinus |
| CN112812198A (en) * | 2021-01-13 | 2021-05-18 | 西藏新浩药业有限公司 | Method for extracting polysaccharide from nettle |
| CN116731222A (en) * | 2023-07-13 | 2023-09-12 | 昆明市延安医院 | Nettle rhamnogalacturonan and preparation method and application thereof |
| CN116731222B (en) * | 2023-07-13 | 2024-01-30 | 昆明市延安医院 | Nettle rhamnogalacturonan and preparation method and application thereof |
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Open date: 20091118 |