CN103588893B - A kind of preparation method of southern Anhui Umbilicaria esculenta polysaccharide - Google Patents
A kind of preparation method of southern Anhui Umbilicaria esculenta polysaccharide Download PDFInfo
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Abstract
The preparation method that the invention discloses a kind of southern Anhui Umbilicaria esculenta polysaccharide。It is with Anhui Province Wan Nan real estate Umbillcaria esculenta for raw material, first Umbillcaria esculenta is pulverized, reusable heat flooding 3～4 times, each lixiviate 2～3h, merge lixiviating solution, lixiviating solution carries out membrance separation and obtains rubble fungus polysaccharides permeate, permeate obtains rubble fungus polysaccharides trapped fluid then through membrance separation, and trapped fluid carries out macroporous adsorbent resin column chromatography again, and eluent concentrates, precipitate with ethanol obtains Umbillcaria esculenta polyose, and its final purity is up to 92.5%。A whole set of production technology continuous operation, energy consumption is low, mild condition, do not destroy effective ingredient, and safety is high, Umbillcaria esculenta polysaccharide yield is high。Resin reproducible utilization；The recyclable recycling of solvent, environmental friendliness, conservation, reduces cost。Provide as the exploitation of medicine resource and health food for southern Anhui Umbilicaria esculenta polysaccharide and established technical foundation。
Description
Technical field
The preparation method that the present invention relates to a kind of southern Anhui Umbilicaria esculenta polysaccharide, belongs to food, processing of farm products and bioengineering field。
Background technology
Umbillcaria esculenta (Umbilicariaesculenta or Umbilicariahoffm) is Umbilicariaceae (Umbilicariaceae, also referred to as Qi Yi section) sporophore of plant, it it is the homobium of algae and fungi, it is born on rock, body is flat, in irregular cycle, above brown, the back side is by black fine hair。Be distributed mainly on the ground such as Anhui, Zhejiang, Jiangxi, there is heat-clearing and toxic substances removing, quench the thirst eliminate the phlegm, antiinflammatory of relievining asthma, improving eyesight benefit essence etc. effect, it is possible to be used as medicine。Containing substantial amounts of polysaccharide, colloid, ferrum, phosphorus, calcium, multivitamin, umbilicaric acid and the acid of Hydrargyri Oxydum Rubrum rock etc. in Umbillcaria esculenta。Pharmacological evaluation shows that the cell of body, body fluid and nonspecific immunity are all had obvious potentiation by Umbillcaria esculenta polysaccharide, and all has good antioxidation, hypoglycemic activity。
At present, have no the patent about Umbillcaria esculenta polyoses producing method, have the research report that a few studies person has been correlated with only。As Wang Weidong etc. with pick up from Tiantangzhai Dabie Mountain, Luotian, Hubei Province main peak Umbillcaria esculenta for raw material, orthogonal experiment method is adopted to filter out lixiviate optimised process: temperature is 80 DEG C, extraction time 3h, solid-liquid ratio is 1: 10, extraction ratio 4.63% (the Wang Weidong of water solublity Umbillcaria esculenta polysaccharide, Wang Hongyun, Yang Li. the Study on extraction of Umbillcaria esculenta polysaccharide. food science and technology, 2010,35 (4): 193-195)。Zhu Shuanjie etc. with pick up from Langyashan Mountain In Chuzhou, Anhui Province Umbillcaria esculenta for raw material, utilize yeast fermentation method come purification Umbillcaria esculenta polysaccharide (Zhu Shuanjie, Xiang Yuyong, Luo Xia etc. yeast fermentation method extract Umbillcaria esculenta polysaccharide process optimize. Beijing Agricultural College's journal, 2012,27 (1): 65-68)。Gu Lizhaer etc. are to adopt murus Umbillcaria esculenta in a number glacier of Xinjiang Tianshan and light skin root Umbillcaria esculenta for raw material, obtain the optimum extraction process of polysaccharide: concentration of alcohol 95%, Extracting temperature 80 DEG C, solid-liquid ratio 1: 20, extraction time 3.0h, its extraction ratio is 15.07% (Gu Lizhaer, buy and carry Haas wood, handkerchief is diligent to be carried. the research of the extraction process of light skin root Umbillcaria esculenta polysaccharide. and food science and technology, 2011,36 (10): 162-165)；And the monosaccharide composition in polysaccharide has been studied, two kinds of water soluble polysaccharide are easily separated purification through DEAE-52 ion exchange column and SephadexG-150 gel column and obtain purified polysaccharide, the monosaccharide of light skin root Umbillcaria esculenta purified polysaccharide consists of mannose, glucose, galactose, and the ratio of its content is 2.97: 17.70: 1.52；Murus Umbillcaria esculenta purified polysaccharide is containing only glucose, content is 5.37%, two kinds of Umbillcaria esculentas have different monosaccharide composition (Gu Lizhaer, nguktrum is than Asia, gram heat wood river etc. in two kinds of Umbillcaria esculentas the extraction of polysaccharide with separate purification and monosaccharide composition analysis thereof. Food Science, 2011,32 (21): 48-51)。
Above-mentioned relevant report and research are all the preliminary extractions that the produced Umbillcaria esculenta in area, south, non-Anhui is carried out, and the purity of the Umbillcaria esculenta polysaccharide of gained is relatively low, and it does not further separate purification；The technique that its laboratory obtains is difficult to amplify and industrialized production；Seriality, complete preparation technology capable of being industrialized and method have no research report。
Separate sources, different genera Umbillcaria esculenta there is different material composition, its poor activity opposite sex is also bigger。Up to now, south, the Anhui Umbillcaria esculenta and the polysaccharide thereof that have antithrombotic acitivity concurrently have no correlational study report。South, Anhui real estate Umbillcaria esculenta has been carried out innovative research by us, result of study shows: southern Anhui Umbilicaria esculenta polysaccharide is mainly made up of mannans polysaccharide, relatively big with the diversity that Umbillcaria esculenta composition is produced such as Wan Bei, Wan Xi in other areas inside the province, rear both are mainly made up of glucan polysaccharide and arabinan class polysaccharide。Additionally, further biological activity test shows: though being all Umbillcaria esculenta Pseudomonas, the difference of kind, molecular structure and chemical composition difference are relatively big, and its using dosage, activity intensity and toxic and side effects there is also larger difference。Being compared by dose,equivalent and timeliness effect, the biological activity of southern Anhui Umbilicaria esculenta polysaccharide is strong, toxic and side effects is relatively low。It can affect normal and rat's blood stasis model hemorheology, it is suppressed that carotid artery thrombosis, it is suppressed that platelet aggregation。It shows powerful anti thrombotic action in the activity test of animal model: after blood stasis model administration, maximum platelet aggregation rate significantly reduces；Can extending the rat-tail bleeding time, effectiveness is slightly below warfarin。And to erythrocyte membrane K+Ion current has inhibitory action, and can make Cl-Ion channel opens, erythrocyte deformability is all improved by these certain impact, reduces whole blood viscosity。
South, Anhui Umbillcaria esculenta aboundresources, distribution is extensively had in area, south, Anhui, with high nutritive value and effect, there is commercial value, it is subject to the attention of numerous Wan Nan private enterprise and carries out large-scale cultivation, different from the Umbillcaria esculenta in other areas composition, southern Anhui Umbilicaria esculenta polysaccharide content is higher, has important value in the edible and exploitation of the medicinal function factor。It mainly contains more than ten kind polysaccharide and other materials, and structure is complicated, it is difficult to extracting and developing and purification。A kind of Umbillcaria esculenta for area, south, Anhui institute special product, our employing is prone to industrialized membrane separation technique and macroporous adsorption resin chromatography method and is has been prepared by south, Anhui real estate Umbillcaria esculenta, obtain its high-purity polysaccharide component (high purity 92.5%), be that production basis has been established in the exploitation as medicine resource and health food for it。
Summary of the invention
It is an object of the invention to provide that a kind of equipment is simple, cost is low, can industrialized production, non-environmental-pollution a whole set of process of preparing of a kind of southern Anhui Umbilicaria esculenta polysaccharide。
The preparation method of a kind of southern Anhui Umbilicaria esculenta polysaccharide, comprises the following steps:
(1) south, Anhui real estate Umbillcaria esculenta is dried and pulverizes；
(2) by the Umbillcaria esculenta after pulverizing with hot water extraction, each time is 2～3 hours, and total immersion carries 3～4 times；
(3) inputting film separation system 1 through excess pressure pump after extracting solution filtration, cooling, cellulose, macro-molecular protein and pigment in membrance separation trapped fluid, containing monosaccharide, polysaccharide, fat, aminoacid, small protein and pigment in membrance separation permeate；
(4) membrance separation permeate inputs film separation system 2 then through compression pump, and membrance separation retains out the Umbillcaria esculenta polysaccharide concentrate containing monosaccharide, fat, aminoacid, small protein and pigment；
(5) Umbillcaria esculenta polysaccharide concentrate carries out macroporous adsorbent resin column chromatography, and with 30%-60% alcoholic solution eluting, eluent concentrates, and adds ethanol to ethanol content 80%, stands and filter, obtain Umbillcaria esculenta polyose；
(6) waste liquid in step (3) and step (4) membrance separation and step (5) macroporous adsorbent resin column chromatography, carries out reclaiming environmental protection treatment；
(7), in step (5) during eluent concentration, ethanol carries out backflow and reclaims；Ethanol in filtrate after recovery filtration。
In described step (2), during lixiviate, the ratio of Umbillcaria esculenta and water is 1: 5～40 (preferably 1: 10～35)；Extraction temperature is 60～90 DEG C。
In described step (3), film separation system 1 retaining molecular weight is 15000 dalton。
In described step (4), film separation system 2 retaining molecular weight is 6000 dalton。
Compression pump operating pressure 0.01～0.2Mpa in described rapid (3) and (4)。
In described step (5), macroporous resin is AB-8, DM130, DA201, X-5, HPD400 or HPD800。
Filtering drainage screen used in described step (5) is 140～250 orders。
A kind of southern Anhui Umbilicaria esculenta polysaccharide, is prepared from according to said method。
Southern Anhui Umbilicaria esculenta polysaccharide prepared by the method for the invention, its purity is more than 90%。
The Advantageous Effects of the present invention:
1, solvent for use has only to second alcohol and water, does not produce any to affect the liquid and waste slag produced of environment, and recyclable；
2, have employed high performance membrane isolation technics and macroporous resin chromatography, energy consumption is low, operating condition is gentle, safety is high, do not destroy the activity of southern Anhui Umbilicaria esculenta polysaccharide, and the resin iterative regenerable of the latter utilizes；
3, the yield of southern Anhui Umbilicaria esculenta polysaccharide and purity are high, and yield may be up to 21.7%, high purity 92.5%；
4, extracting and developing, purification and concentration can a step complete, and a whole set of production technology can continuous operation；
5, equipment is simple, and cost is few, invests little。
Accompanying drawing explanation
Fig. 1 is the preparation technology flow chart of southern Anhui Umbilicaria esculenta polysaccharide。
Detailed description of the invention
Below technical solution of the present invention is described in detail, but protection scope of the present invention is not limited to described embodiment。
The process of preparing of the southern Anhui Umbilicaria esculenta polysaccharide of the present invention, comprises the steps:
(1) south, Anhui real estate Umbillcaria esculenta is dried and pulverizes；
(2) by the Umbillcaria esculenta after pulverizing with hot water extraction, extraction temperature is 60～90 DEG C, and the ratio of Umbillcaria esculenta and water is 1: 5～40 (preferably 1: 10～35), each extraction time is 2～3 hours, filtered filtration residue is lixiviate again, altogether extracting 3～4 times, merges lixiviating solution；
(3) under 0.01～0.2Mpa operating pressure, film separation system 1, cellulose, macro-molecular protein and pigment etc. in membrance separation trapped fluid are inputted through excess pressure pump after lixiviating solution filtration, cooling；Containing monosaccharide, polysaccharide, fat, aminoacid, small protein and pigment etc. in membrance separation permeate；Described macro-molecular protein retaining molecular weight all relative to small protein, is namely macro-molecular protein more than retaining molecular weight, otherwise is small protein。
(4) permeate inputs film separation system 2 then through compression pump under 0.01～0.2Mpa operating pressure, and membrance separation retains out the Umbillcaria esculenta polysaccharide concentrate containing monosaccharide, fat, small protein and pigment etc.；
(5) Umbillcaria esculenta polysaccharide concentrate carries out macroporous adsorbent resin column chromatography, and with obtaining eluent after ethanol (30～60%) eluting, eluent concentrates, and adds ethanol to alcohol content 80%, stands and filter, obtain Umbillcaria esculenta polyose；
(6) waste liquid in membrance separation and macroporous adsorbent resin column chromatography and omission timber, all carries out reclaiming environmental protection treatment；
(7) ethanol that macroporous adsorbent resin column chromatography relates to, all carries out recycling and recycling。During eluent concentration, ethanol is reflowable；Filtering to get filtrate after precipitate with ethanol, ethanol is recyclable；
The retaining molecular weight of described film separation system 1 is 15000 dalton, and the retaining molecular weight of film separation system 2 is 6000 dalton；
Described column chromatography macroporous adsorbent resin is AB-8, DM130, DA201, X-5, HPD400 or HPD800；
Drainage screen used by described filtration is 140～250 mesh filter screens。
The mensuration of southern Anhui Umbilicaria esculenta polysaccharide takes Phenol sulfuric acid procedure, specific as follows:
1, standard curve making。Take the dextrose standard sample 500mg being dried to constant weight at 105 DEG C, distilled water dissolves and is settled to 100mL, take 1.0mL again and be settled to 100mL, shake up to obtain 50 μ g/mL glucose standard, draw 0.2,0.4,0.6,0.8,1.0,1.2,1.4,1.6, the above-mentioned solution of 1.8mL add tool plug test tube, mend distilled water to 2.0mL。Add 6% and heavily steam phenol solution 1.0mL, be rapidly added 5mL concentrated sulphuric acid and shake up, after left at room temperature 30min, under 490nm, survey its absorbance。With standard glucose concentration for abscissa, absorbance is vertical coordinate drawing standard curve, obtains regression equation: y=0.0133+0.0028, its R2=0.9986)。
2, southern Anhui Umbilicaria esculenta polysaccharide measures。Umbillcaria esculenta polysaccharide extraction liquid is diluted to finite concentration, and its scope is in standard curve。Take 1.0mL diluent in test tube, add 1.0mL water, add 6% and heavily steam phenol solution 1.0mL, it is rapidly added 5.0mL concentrated sulphuric acid to shake up, after left at room temperature 30min, under 490nm, surveys its absorbance, obtain the total sugar content of sample according to regression equation, calculating its purity is 89.6～92.5%
Embodiment 1
1, taking south, 1000g Anhui Umbillcaria esculenta, dry in the sun about 3 days, pulverizer is pulverized。
2, adding 10000g water, heat after soaking 2.5h to 80 DEG C, filter with 200 mesh filter screens, filtering residue is lixiviate again, altogether extracting 3 times, merges lixiviating solution。
3, lixiviating solution is cooled to room temperature, and inputting membrane molecule amount under 0.2Mpa operating pressure through compression pump is 15000 daltonian film separation systems。
Cellulose, macro-molecular protein and pigment etc. in membrance separation trapped fluid；Containing monosaccharide, polysaccharide, fat, little molecule aminoacid, small protein and pigment etc. in membrance separation permeate。
4, permeate inputs membrane molecule amount through compression pump under 0.2Mpa operating pressure is 6000 daltonian film separation systems。
Membrance separation retains out the Umbillcaria esculenta polysaccharide concentrate containing monosaccharide, fat, aminoacid, small protein and pigment etc.。
5, Umbillcaria esculenta polysaccharide concentrate AB-8 macroporous adsorbent resin carries out column chromatography, obtains omission timber, and omission timber carries out reclaiming environmental protection treatment；Obtaining eluent after the ethanol elution of 50%, eluent concentrates, and adds ethanol to alcohol content 80%, stands and filter, obtain 189g Umbillcaria esculenta polyose, and yield is 18.9%, and purity is 92.5%。
6, the waste liquid after twice membrance separation all carries out reclaiming environmental protection treatment。
7, the ethanol of eluting AB-8 macroporous adsorbent resin, may be recovered process and recycles。During eluent concentration, ethanol is reflowable；Filtering to get filtrate after precipitate with ethanol, ethanol reclaims。
Embodiment 2
1, taking south, 1000g Anhui Umbillcaria esculenta, dry in the sun about 3 days, pulverizer is pulverized；
2, adding 20000g water, heat after soaking 3h to 90 DEG C, filter with 220 mesh filter screens, filtering residue is lixiviate again, altogether extracting 4 times, merges lixiviating solution；
3, lixiviating solution is cooled to room temperature, and inputting membrane molecule amount under 0.15Mpa operating pressure through compression pump is 15000 daltonian film separation systems, cellulose, macro-molecular protein and pigment etc. in membrance separation trapped fluid；Containing monosaccharide, polysaccharide, fat, little molecule aminoacid, small protein and pigment etc. in membrance separation permeate；
4, permeate inputs membrane molecule amount through compression pump under 0.15Mpa operating pressure is 6000 daltonian film separation systems, and membrance separation retains out the Umbillcaria esculenta polysaccharide concentrate containing monosaccharide, fat, aminoacid, small protein and pigment etc.；
5, Umbillcaria esculenta polysaccharide concentrate DA201 macroporous adsorbent resin carries out column chromatography, obtains omission timber, and omission timber carries out reclaiming environmental protection treatment；Obtaining eluent after the ethanol elution of 60%, eluent concentrates, and adds ethanol to alcohol content 80%, stands and filter, obtain 217g Umbillcaria esculenta polyose, and yield is 21.7%, and purity is 89.6%；
6, the waste liquid after twice membrance separation all carries out reclaiming environmental protection treatment；
7, the ethanol of eluting DA201 macroporous adsorbent resin, may be recovered process and recycles。During eluent concentration, ethanol is reflowable；Filtering to get filtrate after precipitate with ethanol, ethanol reclaims。
Embodiment 3
1, taking south, 1000g Anhui Umbillcaria esculenta, dry in the sun about 3 days, pulverizer is pulverized；
2, adding 30000g water, heat after soaking 3h to 80 DEG C, filter with 200 mesh filter screens, filtering residue is lixiviate again, altogether extracting 3 times, merges lixiviating solution；
3, lixiviating solution is cooled to room temperature, and inputting membrane molecule amount under 0.10Mpa operating pressure through compression pump is 15000 daltonian film separation systems, cellulose, macro-molecular protein and pigment etc. in membrance separation trapped fluid；Containing monosaccharide, polysaccharide, fat, little molecule aminoacid, small protein and pigment etc. in membrance separation permeate；
4, permeate inputs membrane molecule amount through compression pump under 0.10Mpa operating pressure is 6000 daltonian film separation systems, and membrance separation retains out the Umbillcaria esculenta polysaccharide concentrate containing monosaccharide, fat, aminoacid, small protein and pigment etc.；
5, Umbillcaria esculenta polysaccharide concentrate X-5 macroporous adsorbent resin carries out column chromatography, obtains omission timber, and omission timber carries out reclaiming environmental protection treatment；Obtaining eluent after the ethanol elution of 50%, eluent concentrates, and adds ethanol to alcohol content 80%, stands and filter, obtain 201g Umbillcaria esculenta polyose, and yield is 20.1%, and purity is 91.3%；
6, the waste liquid after twice membrance separation all carries out reclaiming environmental protection treatment；
7, the ethanol of eluting X-5 macroporous adsorbent resin, may be recovered process and recycles。During eluent concentration, ethanol is reflowable；Filtering to get filtrate after precipitate with ethanol, ethanol reclaims。
Although as it has been described above, represented and described the present invention with reference to specific preferred embodiment, but it shall not be construed as the restriction to the present invention self。Under the spirit and scope of the present invention premise defined without departing from claims, it can be made in the form and details various change。
Claims (1)
1. the preparation method of a southern Anhui Umbilicaria esculenta polysaccharide, it is characterised in that comprise the following steps:
(1) south, Anhui real estate Umbillcaria esculenta is dried and pulverizes；
(2) by the Umbillcaria esculenta after pulverizing with hot water extraction, during lixiviate, the ratio of Umbillcaria esculenta and water is 1: 5～40；Extraction temperature is 60～90 DEG C, each 2～3 hours, lixiviate 3～4 times；
(3) inputting film separation system 1 through excess pressure pump after extracting solution filtration, cooling, cellulose, macro-molecular protein and pigment in membrance separation trapped fluid, containing monosaccharide, polysaccharide, fat, aminoacid, small protein and pigment in membrance separation permeate；Described film separation system 1 retaining molecular weight is 15000 dalton；Described compression pump operating pressure 0.01～0.2MPa；
(4) membrance separation permeate inputs film separation system 2 then through compression pump, and membrance separation retains out the Umbillcaria esculenta polysaccharide concentrate containing monosaccharide, fat, aminoacid, small protein and pigment；Described film separation system 2 retaining molecular weight is 6000 dalton；Described compression pump operating pressure 0.01～0.2MPa；
(5) Umbillcaria esculenta polysaccharide concentrate carries out macroporous adsorbent resin column chromatography, and with 30%～60% alcoholic solution eluting, eluent concentrates, and adds ethanol to ethanol content 80%, stands and filter, obtain Umbillcaria esculenta polyose；Described macroporous adsorbent resin is AB-8, DM130, DA201, X-5, HPD400 or HPD800；Drainage screen used by described filtration is 140～250 orders；
(6) waste liquid in step (3) and step (4) membrance separation and step (5) macroporous adsorbent resin column chromatography, carries out reclaiming environmental protection treatment；
(7), in step (5) during eluent concentration, ethanol carries out backflow and reclaims；Reclaim ethanol in filtrate。
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| CN101289517A (en) * | 2008-05-30 | 2008-10-22 | 广西师范大学 | Process for abstracting momordica grosvenori polysaccharide |
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| CN101289517A (en) * | 2008-05-30 | 2008-10-22 | 广西师范大学 | Process for abstracting momordica grosvenori polysaccharide |
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