CN101628070B - Chinese yam polysaccharides extract and preparation method and application thereof - Google Patents
Chinese yam polysaccharides extract and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a Chinese yam polysaccharides extract and a preparation method and application thereof. The preparation method comprises the following steps: crushing Chinese yam slices, extracting by ethanol, separating by centrifugation, depositing dregs and adding water, stirring, grinding by a colloid grinder, extracting by water, separating by centrifugation, decompressing and concentrating supernatant to prepare liquid extract; depositing the liquid extract by the ethanol, separating by centrifugation, depositing dregs and adding water, stirring and dissolving, separating by centrifugation, and drying liquid extract to prepare the Chinese yam polysaccharides extract, wherein the yield is 10-19 percent. The Chinese yam polysaccharides extract does not contain starch, and the weight percentage content of the Chinese yam polysaccharides extract is 30-61 percent. The Chinese yam polysaccharides extract has the healthcare function of assisting to regulate the blood sugar level and enhancing the immunity, and can be used for preparing medicines or health care products of regulating the blood sugar level and enhancing the immunity.
Description
Technical field
The invention belongs to the processing extraction and the separation technology field of food, natural product, relate to the polyoses extract of a kind of food, natural drug Rhizoma Dioscoreae.
Background technology
Rhizoma Dioscoreae is the tuber of the perennial voluble herb plant of Dioscoreaceae Rhizoma Dioscoreae Dioscorea opposite Thunb..The effect that Rhizoma Dioscoreae has spleen reinforcing, supports lung, reinforces the kidney, benefit is smart.Shennong's Herbal is classified as top grade." main damaging the spleen and stomach tonify deficiency is won, and removes the cold and heat pathogen, invigorating the spleen and replenishing QI power, longue meat to say it.Clothes are clear-headed and clear-sighted for a long time, not hungry the prolonging life of making light of one's life by commiting suicide." the Compendium of Material Medica cloud: " Rhizoma Dioscoreae sweet in the mouth warm in nature is flat nontoxic, strengthening the spleen and stomach, lung benefiting kidney, stopping leak dysentery, the saliva that reduces phlegm, profit fur ".Cure mainly diarrhea due to hypofunction of the spleen, chronic dysentery, the chronic consumptive disease diseases such as leukorrhagia, frequent micturition of coughing, quench one's thirst, pass out semen.
Chinese yam polysaccharide can improve the lymphocytic multiplication capacity of T, NK cytoactive, active, the serum IgG content of serum hemolysin to some extent, also can strengthen the phagocytic activity of macrophage.Therefore, Chinese yam polysaccharide has tangible antitumor, enhance immunity is regulated and the effect of mutation; Both had non-specific immunity, had the specific cellular immunity of raising and humoral immune function again; Comprehensive body's immunological function of improving is had great importance.
Rhizoma Dioscoreae can be supplied with a large amount of mucus albumen of human body, and this is the complex that polysaccharide combines with protein, and the lipidosis of ability Prevention of Cardiovascular system keeps elasticity of blood vessels, prevents atherosclerotic generation, reduces the subcutaneous fat deposition, avoids occurring obesity.Can also prevent liver and kidney connective tissue atrophy, keep the lubricated of digestive tract, respiratory tract and articular cavity.Mucopolysaccharide in the Rhizoma Dioscoreae with can form sclerotin after inorganic salt combines, strengthen the elasticity of cartilage.
Chinese yam polysaccharide to the treatment of diabetes effect with increase insulin secretion, to improve impaired beta Cell of islet function relevant; Experiment in vitro proves that Rhizoma Dioscoreae also can be used as the drug supporting healthy energy of antitumaous effect, and this possibly have very strong immunoloregulation function relevant (Cheng Lin etc., the extraction process of the preferred Chinese yam polysaccharide of orthogonal experiment method, Chinese medicine and clinical, 2005,5 (9)) with it.From theory of Chinese medical science, diabetes show as deficiency of both QI and YIN, phlegm-turbidity and blood stasis blood, venation block.Rhizoma Dioscoreae is a conventional Chinese medicine, and medicinal history is long, and its main effect is a strengthening the spleen and nourishing the stomach, the lung benefiting that promotes the production of body fluid, the kidney invigorating arresting seminal emission, is one of main medicine of Chinese traditional treatment " diabete ".(Zhang Zhongquan etc., Chinese yam polysaccharide discharges the research that influences, journal of shanghai Chinese medicine, 2003,37 (10) to rat blood sugar and islets of langerhans).
Diabetes are by hypoinsulinism or Insulin receptor INSR is insensitive causes, and are the metabolic disease of principal character with hyperglycemia companion hyperlipidemia, and it can cause the relevant diseases of eye, kidney, nerve, heart and blood vessel.According to World Health Organization (WHO), the whole world has 1.8 hundred million diabeticss approximately, and China just has an appointment 6,000 ten thousand, and wherein 80% is type ii diabetes, and the expectation whole world will break through 300,000,000 by 2025 years.Diabetes become a kind of non-infectious chronic disease of serious harm human health, have become the great public health problem that countries in the world are paid close attention to, also become the main public health problem of China.
Though oral western medicine treatment diabetes have clear and definite hypoglycemic activity, because Western medicine has the shortcoming that significant side effects, inconvenient life-time service etc. are difficult to overcome.Chinese yam polysaccharide has treatment of diabetes can enhancing immunity, and effect that again can auxiliary hyperglycemic is taken for a long time and had no side effect.
Because the diabetes enormous amount, and be a kind of lifelong disease, need to use throughout one's life the health food of hypoglycemic medicine or auxiliary hyperglycemic.So from natural products such as Chinese herbal medicine, food, seek effectively medicine, the health food of treatment diabetes, market prospect is very wide.
Chinese yam polysaccharide was pressed the traditional method of Chinese medicine mostly in the past; Use water boiling and extraction with the Rhizoma Dioscoreae decoction pieces, the extraction ratio of Chinese yam polysaccharide is very low, and has a large amount of Rhizoma Dioscoreae starchs in the Rhizoma Dioscoreae extract; Thereby being difficult to accurately measure the content of polysaccharide in the Rhizoma Dioscoreae extract, product quality is difficult to monitoring.
Except that above-mentioned traditional method, there is document to disclose the method for preparing of Chinese yam polysaccharides extract in the prior art, a kind of method is with Rhizoma Dioscoreae powder decocting, filtration, concentrates; Precipitate with ethanol, centrifugal pure molten part and the precipitate with ethanol part of getting, precipitate with ethanol is partly used water dissolution, the centrifugal insoluble matter of removing; Aqueous solution is with 1%CTAB (cetyl trimethyl bromine ammonium salt) solution precipitation; Supernatant 95% ethanol precipitation, deposition is redeposition through water-soluble back, and lyophilization obtains neutral polysaccharide partly (SUP); The CTAB deposition is with 4mol/L NaCl dissolving, and the centrifugal insoluble matter of removing suitably concentrates the back and uses 95% ethanol precipitation, and deposition obtains acidic polysaccharose position (PPT) through lyophilization, and SUP is through Sephadex G-100 gel chromatography.(extraction separation of Chinese yam polysaccharide and structure determination, natural drug, 2003,1 (3)).Whether this method technology is numerous and diverse, does not report the yield and the content of Chinese yam polysaccharide, do not explain in the Chinese yam polysaccharide starch-containing; With polydextran gel (Sephadex G-100) column chromatography, the polydextran gel material is very expensive, and CTAB (cetyl trimethyl bromine ammonium salt) is poisonous, be not useable for food processing, so this method is difficult to realize suitability for industrialized production.
Another kind of reported in literature is broken into 40 orders with exsiccant Rhizoma Dioscoreae powder, 90 ℃ of defats of petroleum ether 1 time; 90 ℃ of 80% ethanol extract 2 times, and 60～64 ℃ of water extraction 2 times filter; Vacuum concentration adds 95% ethanol (concentration of alcohol is reached more than 70%) and separates out polysaccharide, with absolute ethanol washing repeatedly; Polysaccharide dissolves in water again, and lyophilization promptly gets the Rhizoma Dioscoreae crude polysaccharides.Use Sevage (chloroform+n-butyl alcohol) method to remove the protein in the Rhizoma Dioscoreae crude polysaccharides then.(Rhizoma Dioscoreae water soluble polysaccharide Study on extraction process, food and machinery, 2006,22. (2)).This method Rhizoma Dioscoreae crude polysaccharides content 47～57%, yield 9.5～17%.Because 90 ℃ of defats of this method petroleum ether; 90 ℃ of extractions of 80% ethanol; 60～64 ℃ of water extraction will inevitably mix lot of starch in the Chinese yam polysaccharides extract under such temperature conditions, and are difficult to remove; Cause the determination of polysaccharide data of Rhizoma Dioscoreae crude polysaccharides extract seriously higher, so whether the author does not mention in the Rhizoma Dioscoreae crude polysaccharides starch-containing.With Sevage method Deproteinization, chloroform toxicity is big, is not useable for food processing.Use defat with petroleum ether, inflammable and explosive.This method technology is numerous and diverse, equally also is difficult to realize suitability for industrialized production.
Also have a kind of literature method with the fresh Rhizoma dioscoreae slurry through the petroleum ether defat that refluxes, filter, filtrating is reclaimed petroleum ether.Filtering residue spends the night with 80% alcohol solution dipping, reflux, extract.With the filtering residue adding distil water, 50 ℃ of heat are carried, and filtrate decompression concentrates, and Sevage method deproteinization adds activated carbon decolorizing, and is centrifugal, and filtrating adds 95% ethanol, 4 ℃ of hold over night.Centrifugal, precipitate repeatedly washs with dehydrated alcohol, acetone, absolute ether, and vacuum drying must be made with extra care Chinese yam polysaccharide (extraction process of Rhizoma Dioscoreae crude polysaccharides, food and biotechnology journal, 2006,25 (3)).This method is directly extracted with fresh Rhizoma dioscoreae, is merely for 0.2449% (being about 1.7143% in dried Rhizoma Dioscoreae crude polysaccharides yield) in fresh Rhizoma dioscoreae crude polysaccharides yield, and yield is low, does not disclose the content of Chinese yam polysaccharide; With Sevage method (chloroform+n-butyl alcohol) deproteinization, acetone, absolute ether washing, solvent toxicity is big, inflammable and explosive, and cost is high; With Sevage method deproteinization, also removed the glycoprotein (complex polysaccharide) of biologically active simultaneously; This method technology is numerous and diverse, is difficult to realize suitability for industrialized production equally.
Summary of the invention
The object of the present invention is to provide a kind of method for preparing of Chinese yam polysaccharides extract, can change the method for traditional Rhizoma Dioscoreae decoction pieces, produce Chinese yam polysaccharides extract with water boiling and extraction; It is numerous and diverse to overcome in the prior art the existing technology of Chinese yam polysaccharide method for distilling; Use toxic solvent chloroform and inflammable and explosive solvents such as n-butyl alcohol, acetone, petroleum ether or ether, and used polydextran gel (Sephadex G-100) material shortcoming such as costliness very; Solve and have technical problems such as the low and yield of a large amount of Rhizoma Dioscoreae starchs, Chinese yam polysaccharide content is low in the existing Chinese yam polysaccharides extract.
Another object of the present invention is to provide a kind of not amyloid Chinese yam polysaccharides extract.
The present invention also aims to provide said Chinese yam polysaccharides extract to be used to prepare and have the medicine of blood sugar lowering, enhancing immunity effect or the purposes of health food.
The technical scheme that the present invention adopted is following:
A kind of method for preparing of Chinese yam polysaccharides extract is characterized in that: dried yam slice is ground into the above Rhizoma Dioscoreae powder of 200 orders, with 40%～95% ethanol water of 2～10 times of Rhizoma Dioscoreae powder weight, heating and refluxing extraction; It is that 5～20 times water stirs blended rubber body barreling mill, heating and refluxing extraction that ethanol extract centrifugalize, sediment add weight; The aqueous extract centrifugalize, the supernatant concentrating under reduced pressure makes the fluid extract of 7～20 Baume degrees; Centrifugalize behind the 95% ethanol precipitate with ethanol of 2～10 times of amounts of adding fluid extract volume; Sediment adds water stirring and dissolving, the centrifugalize of 2～10 times of weight; Make Chinese yam polysaccharides extract after the supernatant drying.
Described heating and refluxing extraction, extracting temperature is 40 ℃～60 ℃, 1～5 hour extraction time; Extract operation 1～3 time.
Described concentrating under reduced pressure, its thickening temperature are 50 ℃～70 ℃.
Described drying can adopt a kind of drying means in spray drying, vacuum microwave drying or the vacuum lyophilization.
In dried Rhizoma Dioscoreae, Chinese yam polysaccharides extract product yield is 10～19%.
The invention still further relates to a kind of Chinese yam polysaccharides extract, it is characterized in that adopting method for preparing.
Described Chinese yam polysaccharides extract, its key component are polysaccharide and mineral, protein etc.; Wherein polysaccharide quality percentage composition is 30～61%.
Described Chinese yam polysaccharides extract, not starch-containing.Polyoses extract is negative through the iodine reagent identification, shows wherein not starch-containing.Not amyloid Chinese yam polysaccharides extract, the determination of polysaccharide data accurately and reliably.Do not see described not amyloid Chinese yam polysaccharides extract in the market.
The invention still further relates to described Chinese yam polysaccharides extract be used to prepare have blood sugar lowering, the medicine of enhancing immunity effect or the purposes of health food.Described Chinese yam polysaccharides extract; Its polyoses content is high, has the health care of significant auxiliary hyperglycemic, enhancing immunity, and in the preparation process, does not use toxic solvent; Product Safety is high, can be used for preparing have blood sugar lowering, the medicine of enhancing immunity effect, health food.
The method for preparing of Chinese yam polysaccharides extract of the present invention, compared with prior art, Chinese yam polysaccharide content is high, and product yield is high, and production technology is simplified, and can realize suitability for industrialized production.Method of the present invention and prepared product also have following technique effect:
1, method for preparing of the present invention is not used poisonous, inflammable, explosive equal solvent, and production security is high, and Product Safety is high;
2, method for preparing of the present invention, its production process does not produce pollutant such as harmful waste water, waste residue, waste gas;
3, its polyoses content of Chinese yam polysaccharides extract of the present invention is high, has the health care of significant auxiliary hyperglycemic, enhancing immunity, can be used for industries such as medicine, health food.
Describe the present invention below in conjunction with the specific embodiment.Scope of the present invention is not exceeded with the specific embodiment, but is limited the scope of claim.
The specific embodiment
Embodiment 1
Yam slice is ground into the above Rhizoma Dioscoreae powder of 200 orders; 80% ethanol water 250L adds in the 500L reaction pot, adds Rhizoma Dioscoreae powder 50kg again, heating and refluxing extraction 2 hours, 50 ℃ of temperature after starting stirring; Centrifugalize, sediment add 250L water stirs, and grinds with colloid mill and processes the Rhizoma Dioscoreae slurry; With in the Rhizoma Dioscoreae slurry input extraction pot, add 500L water with pump, started the agitating heating reflux, extract, 3 hours, 50 ℃ of temperature; The aqueous extract centrifugalize, 60 ℃ of concentrating under reduced pressure of supernatant make Baume degrees and are 7 fluid extract; Fluid extract adds 95% ethanol precipitate with ethanol of 2～10 times of amounts of volume, alcohol deposit fluid centrifugalize; Sediment adds the water stirring and dissolving of 2～10 times of weight, centrifugalize; Supernatant vacuum lyophilization obtains Chinese yam polysaccharides extract 6.4kg; In dried Rhizoma Dioscoreae, product yield 12.80%.
Prepared Chinese yam polysaccharides extract, its key component are polysaccharide 60.63%, also contain components such as mineral, protein.
Prepared Chinese yam polysaccharides extract, not starch-containing.
Getting concentration is 5 of 0.06% Chinese yam polysaccharides extract aqueous solutions, places white plaque, adds constant blueness behind 1 iodine reagent, i.e. polyoses extract iodine reagent identification is negative, shows wherein not starch-containing.With 0.06% soluble starch as contrast.
Instance 2
Yam slice is ground into the above Rhizoma Dioscoreae powder of 200 orders; 85% ethanol water 500L adds in the 1000L reaction pot, adds Rhizoma Dioscoreae powder 100kg again, heating and refluxing extraction 4 hours, 40 ℃ of temperature after starting stirring; Centrifugalize, sediment add 500L water stirs, and grinds with colloid mill and processes the Rhizoma Dioscoreae slurry; With in the Rhizoma Dioscoreae slurry input extraction pot, add 1000L water with pump, started the agitating heating reflux, extract, 4 hours, 40 ℃ of temperature; The aqueous extract centrifugalize, 70 ℃ of concentrating under reduced pressure of supernatant make Baume degrees and are 20 fluid extract; Fluid extract adds 95% ethanol precipitate with ethanol of 2～10 times of amounts of volume, alcohol deposit fluid centrifugalize; Sediment adds the water stirring and dissolving of 2～10 times of weight, centrifugalize; The supernatant spray drying makes Chinese yam polysaccharides extract 10.8kg; In dried Rhizoma Dioscoreae, product yield 10.80%.
Prepared Chinese yam polysaccharides extract, its key component are polysaccharide 36.61%, also contain components such as mineral, protein.Prepared Chinese yam polysaccharides extract, not starch-containing.
Instance 2 Chinese yam polysaccharides extracts of the present invention prove the health care with significant auxiliary hyperglycemic through the animal function test.
1 material
1.1 the Chinese yam polysaccharides extract of given the test agent: embodiment 2 gained is the pale powder shape.
1.2 experimental animal: select cleaning level BI/F for use
1For healthy male mice, provide by Shanghai Slac Experimental Animal Co., Ltd.
1.3 dosage: establish three dose groups, i.e. low dosage, middle dosage, high dose group, other establishes a matched group.Given the test agent is prepared with sterilized water, high, medium and low dosage, and matched group replaces given the test agent with sterilized water, and the mouse stomach amount is 10mL/kg.bw.
2. test method
The auxiliary hyperglycemic function method of inspection by " health food check and assessment technique standard " (version in 2003) is carried out.
2.1 zoic model with hyperglycemia test
The first phase is made the phase for the mice diabetes model
Select the BI/F of body weight 24～26g for use
1For healthy male mice, after laboratory was quarantined 3, fasting 24h; Water restriction not, with freshly prepared alloxan with 160mg/kg.bw dosage lumbar injection, 1 week back fasting 5h; Water restriction is surveyed fasting glucose, with blood glucose 10～25mmol/L as the hyperglycemia model mice.
The second phase is hyperglycemia model mice blood sugar lowering experimental period
Select 80 of hyperglycemia model mices for use, be divided into 8 groups at random according to the level of blood glucose value, 10 every group, wherein to organize as I for 4 groups, dosage is respectively matched group, low dosage, middle dosage, high dose group, carries out fasting glucose and measures; In addition 4 groups as II, and dosage is respectively matched group, low dosage, middle dosage, high dose group, carries out carbohydrate tolerance test.Per os gives the given the test agent of mice corresponding dosage once a day, and matched group replaces given the test agent with sterilized water, continuous 1 month.Carry out mice fasting glucose and carbohydrate tolerance test during off-test.
2.2 index and method
Under the state of not anaesthetizing, gather mouse orbit angular vein blood, carry out following index determining.
2.2.1 fasting glucose is measured
Mice fasting 5h, water restriction is not gathered eye socket angular vein blood, gets serum and carries out blood sugar detection with the glucose oxidase-peroxidase method with 721 spectrophotometers.Under the prerequisite that model is set up, given the test agent dose groups and matched group compare, and the fasting glucose measured value reduces or blood glucose decline percentage rate has statistical significance, and it is positive that the fasting glucose experimental result falls in this given the test agent of decidable.
2.3.2 carbohydrate tolerance test
Mice fasting 5h, water restriction not, dose groups gives the variable concentrations given the test agent; Model control group gives with the volume solvent; Per os gives glucose 2.0g/kg after 20 minutes, measure to give behind the glucose 0,0.5 respectively, the blood glucose value of 2h, the same 2.1.1 of determination method of blood sugar.Under the prerequisite that model is set up, given the test agent dose groups and matched group relatively, area reduces after giving glucose 0,0.5, under the 2h blood glucose curve has statistical significance, this given the test agent carbohydrate tolerance experimental result positive of decidable.
2.4. the result judges:
Index positive in fasting glucose and the carbohydrate tolerance binomial index, i.e. this given the test agent auxiliary hyperglycemic function zoopery of decidable result is positive.
3 results
3.1 the influence of the hyperglycemia model mouse blood sugar that Chinese yam polysaccharides extract causes alloxan
3.1.1 Chinese yam polysaccharides extract is seen table 1 to the result that influences of the hyperglycemia model mice fasting glucose that alloxan causes:
Table 1 Chinese yam polysaccharides extract is to the influence of hyperglycemia model mice fasting glucose (mmol/L)
| Dose groups | Number of animals (only) | Before the test | The P value | After the test | The P value | The decline percentage rate | The P value |
| Contrast | 10 | 18.01±5.65 | 19.43±4.13 | -17.89±41.09 | |||
| Low dosage | 10 | 18.17±5.75 | 1.000 | 20.65±5.80 | 0.901 | -21.90±42.64 | 0.993 |
| Middle dosage | 10 | 18.01±5.59 | 1.000 | 20.18±4.96 | 0.973 | -22.40±51.05 | 0.990 |
| High dose | 10 | 17.91±5.52 | 1.000 | 14.04±4.59 | 0.049 | -17.60±27.81 | 0.155 |
Visible by table 1, fasting glucose low dosage, middle dosage, high dose group are compared the difference not statistically significant before the test of hyperglycemia model mouse blood sugar with matched group 0mg/kg.The preceding fasting glucose of test that is mice is comparatively balanced between each group.
Per os gives the Chinese yam polysaccharides extract 1 month of mice various dose; Each dose groups mice fasting glucose and the blood glucose decline percentage rate measured carry out homogeneity test of variance; Satisfy the homogeneity of variance requirement, carry out statistical disposition with the comparative approach in twos of mean between a plurality of experimental grouies in the one factor analysis of variance method and matched group.Significantly be lower than matched group by fasting glucose behind the visible high dose group mouse test of table 1 result.
3.1.2 Chinese yam polysaccharides extract is seen table 2 to the result that influences of the hyperglycemia model mice carbohydrate tolerance that alloxan causes:
Table 2 Chinese yam polysaccharides extract is to the influence of hyperglycemia model mice carbohydrate tolerance
Per os gives the Chinese yam polysaccharides extract 1 month of mice various dose; Each dose groups hyperglycemia model mouse blood sugar TG-AUC of being measured carries out homogeneity test of variance; Satisfy the homogeneity of variance requirement, carry out statistical disposition with the comparative approach in twos of mean between a plurality of experimental grouies in the one factor analysis of variance method and matched group.Significantly be lower than matched group by the visible high dose group mouse blood sugar TG-AUC of table 2 result.
4. conclusion
The hyperglycemia model mice that causes with alloxan has been estimated the effect of Chinese yam polysaccharides extract auxiliary hyperglycemic.Chinese yam polysaccharides extract gives hyperglycemia model mice 1 month continuously with low dosage, middle dosage, high dose.The result shows behind the high dose group mouse test that area significantly is lower than matched group under the fasting glucose and blood glucose curve.Show that Chinese yam polysaccharides extract has the function of auxiliary hyperglycemic.
Instance 2 Chinese yam polysaccharides extracts of the present invention are through the animal function test, and also proof has the health care of significant enhancing immunity.
1.ConA inductive mouse spleen lymphocyte conversion test result shows Chinese yam polysaccharides extract when middle dose groups, high dose group, has the effect that strengthens cellular immune function.The result sees table 3:
Table 3 Chinese yam polysaccharides extract is to the influence of the inductive mouse spleen lymphocyte conversion test of ConA
| Group | Number of animals (only) | Add ConA hole and the difference that does not add ConA hole absorbance |
| 0 | 10 | 0.028±0.005 |
| Low dosage | 10 | 0.032±0.005 |
| Middle dosage | 10 | 0.040±0.011* |
| High dose | 10 | 0.042±0.014* |
* Dunnett ' s t check is with negative control group comparing difference significance (P＜0.05)
2. dinitrofluorobenzene inducing mouse delayed allergy test (ear swelling method) result shows Chinese yam polysaccharides extract when middle dose groups, high dose group, has the effect that strengthens cellular immune function.The result sees table 4:
Table 4 Chinese yam polysaccharides extract is to the influence (means standard deviation) of dinitrofluorobenzene inducing mouse delayed allergy
| Group | Number of animals (only) | Auricle weight difference (mg) |
| 0 | 10 | 11.3±2.7 |
| Low dosage | 10 | 13.4±2.1 |
| Middle dosage | 10 | 17.1±2.8* |
| High dose | 10 | 18.5±2.5* |
* Dunnett ' s t check is with negative control group comparing difference significance (P＜0.05)
3. antibody-producting cell detection test (Jerne improves slide method) result shows Chinese yam polysaccharides extract when middle dose groups, high dose group, has the effect that strengthens humoral immune function.The result sees table 5:
Table 5 Chinese yam polysaccharides extract is to the influence (means standard deviation) of hemolysis plaque number
| Group | Number of animals (only) | Hemolysis plaque number (individual/10 6Splenocyte) |
| 0 | 10 | 187±26 |
| Low dosage | 10 | 197±22 |
| Middle dosage | 10 | 217±20* |
| High dose | 10 | 239±14* |
* Dunnett ' s t check is with negative control group comparing difference significance (P＜0.05)
4. the mensuration of serum hemolysin (blood clotting method) result shows Chinese yam polysaccharides extract when high dose group, has the effect that strengthens humoral immune function.The result sees table 6:
Table 6 Chinese yam polysaccharides extract is to the influence (means standard deviation) of mice serum hemolytic antibody product
| Group | Number of animals (only) | The antibody product |
| 0 | 10 | 60.4±4.1 |
| Low dosage | 10 | 67.0±7.4 |
| Middle dosage | 10 | 77.3±5.9 |
| High dose | 10 | 82.5±8.4* |
* Dunnett ' s t check is with negative control group comparing difference significance (P＜0.05)
5. mice carbon clearance test result shows that each dose groups mouse monokaryon phagocyte phagocytic activity of Chinese yam polysaccharides extract is consistent with negative control group.The result sees table 7:
Table 7 Chinese yam polysaccharides extract is to the influence (means standard deviation) of mice phagocytic index
| Group | Number of animals (only) | Phagocytic index |
| 0 | 10 | 4.06±0.61 |
| Low dosage | 10 | 4.43±0.65 |
| Middle dosage | 10 | 4.44±0.76 |
| High dose | 10 | 4.86±0.55* |
* Dunnett ' s t check is with negative control group comparing difference significance (P＜0.05)
6. Turnover of Mouse Peritoneal Macrophages is engulfed the chicken red blood cell result of the test and is shown Chinese yam polysaccharides extract when middle dose groups, high dose group, has the effect that strengthens macrophage phagocytosis of mice.The result sees table 8:
Table 8 Chinese yam polysaccharides extract is to Turnover of Mouse Peritoneal Macrophages
Engulf the influence (means standard deviation) of the phagocytic rate and the phagocytic index of chicken red blood cell
| Group | Number of animals (only) | Phagocytic rate (%) | Phagocytic index |
| 0 | 10 | 18.0±2.2 | 0.21±0.03 |
| Low dosage | 10 | 18.8±2.6 | 0.23±0.04 |
| Middle dosage | 10 | 23.4±1.9* | 0.29±0.03* |
| High dose | 10 | 30.4±3.2* | 0.39±0.04* |
* Dunnett ' s t check is with negative control group comparing difference significance (P＜0.05)
7.NK cytoactive determination test (determination of lactate dehydrogenase method) result shows Chinese yam polysaccharides extract when middle dose groups, high dose group, the effect with enhanced NK cytoactive.The result sees table 9:
Table 9 Chinese yam polysaccharides extract is to the influence (means standard deviation) of NK cytoactive
| Group | Number of animals (only) | NK cytoactive (%) |
| 0 | 10 | 31.8±2.8 |
| Low dosage | 10 | 32.3±2.8 |
| Middle dosage | 10 | 36.8±2.4* |
| High dose | 10 | 47.5±5.6* |
* Dunnett ' s t check is with negative control group comparing difference significance (P＜0.05)
8. conclusion:
The inductive mouse spleen lymphocyte conversion test of the Chinese yam polysaccharides extract ConA result that the inventive method is processed shows when middle dose groups, high dose group, to have the effect that strengthens the mouse cell immunologic function; Dinitrofluorobenzene inducing mouse delayed allergy result of the test shows when middle dose groups, high dose group, to have the effect that strengthens the mouse cell immunologic function; Antibody-producting cell detects result of the test and shows, when middle dose groups, high dose group, has the effect that strengthens the mouse humoral immune function; The mensuration result of serum hemolysin shows when middle dose groups, high dose group, to have the effect that strengthens the mouse humoral immune function; The clearance test of mice carbon is the result show, each dose groups mouse monokaryon phagocyte phagocytic activity is consistent with negative control group; Turnover of Mouse Peritoneal Macrophages is engulfed the chicken red blood cell result of the test and is shown, when middle dose groups, high dose group, has the effect that strengthens macrophage phagocytosis of mice; NK cytoactive determination test (determination of lactate dehydrogenase method) result shows when middle dose groups, high dose group, to have the effect of enhanced NK cytoactive.This Chinese yam polysaccharides extract has the health care of enhancing immunity, can be used as the health food with enhancing immunity.
Claims (8)
1. the method for preparing of a Chinese yam polysaccharides extract is characterized in that: dried yam slice is ground into the above Rhizoma Dioscoreae powder of 200 orders, with 40%～95% ethanol water of 2～10 times of Rhizoma Dioscoreae powder weight, heating and refluxing extraction; It is that 5～20 times water stirs blended rubber body barreling mill, heating and refluxing extraction that ethanol extract centrifugalize, sediment add weight; The aqueous extract centrifugalize, the supernatant concentrating under reduced pressure makes the fluid extract of 7～20 Baume degrees; Centrifugalize behind the 95% ethanol precipitate with ethanol of 2～10 times of amounts of adding fluid extract volume; Sediment adds water stirring and dissolving, the centrifugalize of 2～10 times of weight; Make Chinese yam polysaccharides extract after the supernatant drying.
2. method for preparing according to claim 1 is characterized in that: described heating and refluxing extraction, and extracting temperature is 40 ℃～60 ℃, 1～5 hour extraction time; Extract operation 1～3 time.
3. method for preparing according to claim 1 is characterized in that: described concentrating under reduced pressure temperature is 50 ℃～70 ℃.
4. method for preparing according to claim 1 is characterized in that: described dry adopt spray drying, vacuum microwave drying or vacuum lyophilization.
5. the Chinese yam polysaccharides extract of a method according to claim 1 preparation.
6. Chinese yam polysaccharides extract according to claim 5 is characterized in that: polysaccharide quality percentage composition is 30%～61% in the extract.
7. Chinese yam polysaccharides extract according to claim 5 is characterized in that: described Chinese yam polysaccharides extract is not starch-containing.
The described Chinese yam polysaccharides extract of claim 5 be used to prepare have blood sugar lowering, the medicine of enhancing immunity effect or the purposes of health food.
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