CN102827299A - Efficient extraction and purification method for kumquat polysaccharide - Google Patents
Efficient extraction and purification method for kumquat polysaccharide Download PDFInfo
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- CN102827299A CN102827299A CN2012103073382A CN201210307338A CN102827299A CN 102827299 A CN102827299 A CN 102827299A CN 2012103073382 A CN2012103073382 A CN 2012103073382A CN 201210307338 A CN201210307338 A CN 201210307338A CN 102827299 A CN102827299 A CN 102827299A
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- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 75
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- 238000000034 method Methods 0.000 title claims abstract description 50
- 238000000605 extraction Methods 0.000 title claims abstract description 21
- 238000000746 purification Methods 0.000 title claims abstract description 18
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- 230000035614 depigmentation Effects 0.000 claims abstract description 4
- 241001672694 Citrus reticulata Species 0.000 claims description 101
- 239000002904 solvent Substances 0.000 claims description 29
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 25
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 24
- 239000010931 gold Substances 0.000 claims description 23
- 229910052737 gold Inorganic materials 0.000 claims description 23
- 230000008569 process Effects 0.000 claims description 20
- 238000010828 elution Methods 0.000 claims description 18
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- 239000011780 sodium chloride Substances 0.000 claims description 12
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Landscapes
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Preparation Of Fruits And Vegetables (AREA)
Abstract
本发明公开了一种金柑多糖的高效提取纯化方法，将新鲜金柑榨汁，采用微波、超声波耦合动态逆流浸提技术进行浸提，经离心过滤得到金柑水提液，再采用酶法结合Sevag法脱蛋白、大孔吸附树脂脱色素得到金柑多糖溶液，经DEAESepharoseCL-6B离子交换柱、SephacrylS-400HR凝胶柱分离，再经超滤、减压浓缩、醇沉、冷冻干燥、粉碎制得金柑多糖的精制干粉。本发明首次从金柑中提取多糖，提取时间短，提取率高，并对金柑多糖进行精制，为充分利用我国金柑资源优势和开发应用金柑多糖提供了基础。The invention discloses a high-efficiency extraction and purification method of kumquat polysaccharides. Fresh kumquats are squeezed for juice, extracted by microwave and ultrasonic coupling dynamic countercurrent extraction techniques, centrifuged to obtain kumquat water extracts, and then enzymatically combined with the Sevag method. Deproteinization and macroporous adsorption resin depigmentation to obtain kumquat polysaccharide solution, separated by DEAESepharoseCL-6B ion exchange column and SephacrylS-400HR gel column, then ultrafiltration, vacuum concentration, alcohol precipitation, freeze-drying, and crushing to obtain kumquat polysaccharide refined dry powder. The invention extracts polysaccharides from kumquats for the first time, has short extraction time and high extraction rate, and refines kumquat polysaccharides, which provides a basis for fully utilizing the advantages of kumquat resources in my country and developing and applying kumquat polysaccharides.
Description
Technical field
The invention belongs to food processing technology field, be specifically related to a kind of method for efficiently extracting and purifying of golden mandarin orange polysaccharide.
Background technology
The gold mandarin orange (
Fortunella margarita (Lour.) Swingle) have another name called cumquat, green tangerine, be one of famous special product fruit of China, have abundant nutritive value and special medicinal health effect.The traditional Chinese medical science thinks, golden mandarin orange has been eaten raw and regulated the flow of vital energy, bowl spares, Xie Yu, help digestion, loose and effect such as tremble with fear, reduce phlegm, sober up, and can be used for treating diseases such as pent-up uncomfortable in chest, drunk thirsty, maldigestion, poor appetite, cough, asthma.At present, golden mandarin orange is main to eat raw still, the also rarely seen golden hesperidium aurantium vinegar of rough-wrought product, golden hesperidium aurantium dried meat etc., and added value of product is low.Have the basic research work of customs gold unit mandarin orange still relatively weaker, deep process technology and Products Development lag behind, and have restricted the development of golden mandarin orange industry.
Polysaccharide is the polymer that is formed by connecting monose, extensively is present in animal cell membrane, plant and the microbial cell wall, is except that nucleic acid and another type of important biomolecules the protein in the organism.Big quantity research shows that the function assessment characteristic of polysaccharide is varied; Not only can be used as material for thermal energy storage; And have many biological activitys, like immunomodulatory, antitumor, hypoglycemic, reducing blood-fat, radioprotective, delay senility etc., and to the almost non-toxic spinoff of body.Gold mandarin orange polysaccharide is one of main active ingredient of golden mandarin orange, but has the high efficiency extraction of customs gold unit mandarin orange polysaccharide and the research of separation and purification also not to appear in the newspapers as yet.Therefore, efficiently extracting polysaccharide from golden mandarin orange how obtains high purity, active strong golden mandarin orange polysaccharide is significant.
The process for extracting of polysaccharide is a lot; Process for extracting such as the microwave-assisted extraction of developing in recent years, intensified by ultrasonic wave extraction, high voltage pulse method, dynamic countercurrent lixiviate; Have the efficient height, simple to operate, energy consumption is little; And can not introduce impurity, purity of polysaccharide advantages of higher, in polysaccharide extracts, be used widely.Have the extraction that multiple technologies is combined to be applied to polysaccharide at present,, improve polysaccharide extract rate with the advantage of utilizing various technology like UW countercurrent extraction technology, microwave counter current extractive technique.The method of polysaccharide purification has precipitation classification, Sevag method, chromatography column method, membrane separation process etc., effective neutral component and acidic components in the crude polysaccharide of ion exchange chromatography wherein, and gel permeation chromatography can further be realized the separation of polysaccharides of different molecular weight.Make full use of various extraction purification techniques, it is main purpose of the present invention that a kind of high efficiency extraction of golden mandarin orange polysaccharide and the method for purifying are provided.
Summary of the invention
The object of the present invention is to provide a kind of method for efficiently extracting and purifying of golden mandarin orange polysaccharide; Obtain the golden mandarin orange polysaccharide that extraction time is short, solvent consumption is few, purity is high, activity is strong; To fill up the blank of golden mandarin orange polysaccharide high efficiency extraction and stripping technique; Enrich the carbohydrate science, for the Application and Development of golden mandarin orange polysaccharide provides the basis, promote golden mandarin orange industrial value, expand golden mandarin orange further technological processing way.
For realizing above-mentioned purpose, the present invention adopts following technical scheme:
A kind of method for efficiently extracting and purifying of golden mandarin orange polysaccharide; Fresh golden mandarin orange is squeezed the juice; Adopt microwave, UW Coupled Dynamic adverse current microwave leaching technology to carry out lixiviate; Obtain golden mandarin orange water extract through centrifuging; Adopt enzyme process to combine Sevag method deproteinated, macroporous adsorbent resin depigmentation to obtain golden mandarin orange polysaccharide soln again, separate, make the refining dry powder of golden mandarin orange polysaccharide again through ultrafiltration, concentrating under reduced pressure, alcohol precipitation, lyophilize, pulverizing through DEAE Sepharose CL-6B ion exchange column, Sephacryl S-400 HR gel column.
The concrete steps of said method are:
(1) the fresh golden mandarin orange of screening is through cleaning, stoning, squeeze the juice, subsequent use;
(2) golden mandarin orange juice is quantitatively sent into microwave extraction chamber by the liquid material than 10:1 ~ 30:1 by an endfeed mouth, microwave power 400 ~ 620W, microwave treatment time 15 ~ 30min;
(3) at the UW digester roIl, pure water is injected through under meter control amount by the other end fluid inlet, and solvent flows to discharge port from fluid inlet; When treating that flow of solvent is stablized; Material helical feed in the step (2) is gone into the UW digester roIl, and the liquid material is than being 20:1 ~ 30:1, ultrasonic power 160 ~ 250W; UW 20 ~ 45min action time; Under the promotion of screw, solvent and the formation of golden mandarin orange flow to opposite continuous dynamic countercurrent and carry out the thorough mixing extraction, extract the gained vat liquor and get into next step operation by discharge port;
(4) the centrifugal insoluble impurities of removing of the vat liquor of step (3) obtains golden mandarin orange water extract;
(5) with the golden mandarin orange water extract of step (4), add 40 ℃ of enzymolysis 60 ~ 80min of proteolytic enzyme, adopt Sevag method deproteinated then, centrifugal behind the deproteinated 3 times, obtain supernatant;
(6) with the supernatant of step (5), to cross macroporous adsorbent resin and decolour, selected macroporous adsorbent resin is D101, obtains golden mandarin orange polysaccharide soln after the decolouring;
(7) with the golden mandarin orange polysaccharide soln of step (6); The DEAE Sepharose CL-6B ion exchange column that overbalance is good carries out separation and purification; NaCl solution with zero(ppm) water and 0.1 ~ 1mol/L carries out stepwise elution; Follow the tracks of the effusive component of detection with the phenol sulfuric acid process, collect elution peak respectively, obtain elutriant;
(8) with the elutriant of step (7); The Sephacryl S-400 HR gel-filtration column that overbalance is good carries out further separation and purification, carries out wash-out with the NaCl solution of 0.1 ~ 1mol/L, follows the tracks of with the phenol sulfuric acid process and detects effusive component; Collect elution peak respectively, obtain elutriant;
(9) elutriant with step (8) concentrates through the hollow fiber ultrafiltration membrane desalination and with it respectively, and its molecular weight cut-off is 5000Dal, and WP is controlled at 0.2MPa ~ 0.7MPa, and 20 ~ 60 ℃ of temperature obtain golden mandarin orange polysaccharide soln;
(10) step (9) gained gold mandarin orange polysaccharide is concentrated into 3/10 ~ 2/5 of original volume, adds massfraction and be 80% ethanolic soln alcohol precipitation and spend the night, after the lyophilize of gained deposition, pulverized 60 mesh sieves, promptly obtain the refining dry powder of golden mandarin orange polysaccharide.
Beneficial effect of the present invention:
(1) the present invention adopts microwave ultrasonic wave Coupled Dynamic adverse current microwave leaching technology to extract golden mandarin orange polysaccharide, gives full play to the advantage of microwave technology, ultrasonic technology, dynamic countercurrent microwave leaching technology.Microwave is stronger to the solid penetration, and material is cell rupture under the effect of microwave, and effective constituent is diffused in the solvent apace in the cell.After the microwave extraction, material is admitted to the ultrasonic extraction chamber, and the cavitation effect that the unique mechanical disintegration of UW forms during with oscillating action and through liquid further destroys the material histocyte, the penetration power of increase solvent.The dynamic countercurrent lixiviate makes full use of the biphase concentration gradient continuously, the quick balance of polysaccharide in material and the solvent in the realization leaching process.This invention has the extraction yield height, practices thrift solvent load, characteristic of low energy consumption, and golden mandarin orange polysaccharide extract rate＞10% can be realized the continuity extraction in enormous quantities of golden mandarin orange polysaccharide, is suitable for suitability for industrialized production.
(2) the present invention adopts enzyme process to combine Sevag method deproteinated; The macroporous adsorbent resin depigmentation; And combine DEAE Sepharose CL-6B ion exchange chromatography, Sephacryl S-400 HR gel-filtration column to carry out further separation and purification, obtain purified gold mandarin orange polysaccharide, its purity＞90%.
(3) entire operation process temperature of the present invention is lower, can keep golden mandarin orange active polysaccharide to greatest extent, and to the golden mandarin orange polysaccharide of further research structure activity relationship, it is significant to make full use of China's gold mandarin orange resources advantage Application and Development gold mandarin orange polysaccharide.
Embodiment
A kind of method for efficiently extracting and purifying of golden mandarin orange polysaccharide, concrete steps are following:
(1) the fresh golden mandarin orange of screening is through cleaning, stoning, squeeze the juice, subsequent use;
(2) golden mandarin orange juice is quantitatively sent into microwave extraction chamber by the liquid material than 10:1 ~ 30:1 by an endfeed mouth, microwave power 400 ~ 620W, microwave treatment time 15 ~ 30min;
(3) at the UW digester roIl, solvent is injected through under meter control amount by the other end fluid inlet, and solvent flows to discharge end from the liquid feeding end; When treating that flow of solvent is stablized; Material helical feed in the step (2) is gone into the UW digester roIl, and the liquid material is than being 20:1 ~ 30:1, ultrasonic power 160 ~ 250W; UW 20 ~ 45min action time; Under the promotion of screw, solvent and the formation of golden mandarin orange flow to opposite continuous dynamic countercurrent and carry out the thorough mixing extraction, extract the gained vat liquor and get into next step operation by discharge port;
(4) the centrifugal insoluble impurities of removing of vat liquor in the step (3) obtains golden mandarin orange water extract;
(5) with gained gold mandarin orange water extract in the step (4), add 40 ℃ of enzymolysis 60 ~ 80min of proteolytic enzyme, adopt Sevag method deproteinated then, centrifugal behind the deproteinated 3 times, obtain supernatant;
(6) with step (5) gained supernatant, to cross macroporous adsorbent resin and decolour, selected macroporous adsorbent resin is D101, obtains golden mandarin orange polysaccharide soln after the decolouring;
(7) with step (6) gained gold mandarin orange polysaccharide soln; The DEAE Sepharose CL-6B ion exchange column that overbalance is good carries out separation and purification; Carry out stepwise elution with zero(ppm) water, 0.1 ~ 1mol/L NaCl; Follow the tracks of the effusive component of detection with the phenol sulfuric acid process, collect elution peak respectively, obtain elutriant;
(8) with step (7) gained elutriant; The Sephacryl S-400 HR gel-filtration column that overbalance is good carries out further separation and purification, carries out wash-out with 0.1mol/L NaCl, follows the tracks of with the phenol sulfuric acid process and detects effusive component; Collect elution peak respectively, obtain elutriant;
(9) elutriant in the step (8) is concentrated through the hollow fiber ultrafiltration membrane desalination and with it respectively, its molecular weight cut-off is 5000Dal (WP is controlled at 0.2MPa ~ 0.7Mpa, 20 ~ 60 ℃ of temperature), obtains golden mandarin orange polysaccharide soln;
(10) step (9) gained gold mandarin orange polysaccharide is concentrated into 3/10 ~ 2/5 of original volume, adds 80% ethanol alcohol precipitation and spend the night, after the lyophilize of gained deposition, pulverized 60 mesh sieves, promptly obtain fine gold mandarin orange polysaccharide dry powder.
Below be several specific embodiment of the present invention, further specify the present invention, but the present invention is not limited only to this enforcement.
Embodiment 1
(1) the fresh golden mandarin orange of screening is through cleaning, stoning, squeeze the juice, subsequent use;
(2) with golden mandarin orange juice by an endfeed mouth by the liquid material than the quantitative microwave extraction chamber of sending into of 10:1, microwave power microwave power 450W, microwave treatment time 20min;
(3) at the UW digester roIl, solvent is injected through under meter control amount by the other end fluid inlet, and solvent flows to discharge end from the liquid feeding end; When treating that flow of solvent is stablized, material helical feed in the step (2) is gone into the UW digester roIl, the liquid material is than being 25:1; Ultrasonic power 220W, UW 20min action time is under the promotion of screw; Solvent and the formation of golden mandarin orange powder flow to opposite continuous dynamic countercurrent and carry out the thorough mixing extraction, extract the gained vat liquor and get into next step operation by discharge port;
(4) the centrifugal insoluble impurities of removing of vat liquor in the step (3) obtains golden mandarin orange water extract;
(5) with gained gold mandarin orange water extract in the step (4), add 40 ℃ of enzymolysis 60min of proteolytic enzyme, adopt Sevag method deproteinated then, centrifugal behind the deproteinated 3 times, obtain supernatant;
(6) with step (5) gained supernatant, to cross macroporous adsorbent resin and decolour, selected macroporous adsorbent resin is D101, obtains golden mandarin orange polysaccharide soln after the decolouring;
(7) with step (6) gained gold mandarin orange polysaccharide soln; The DEAE Sepharose CL-6B ion exchange column that overbalance is good carries out separation and purification; Carry out stepwise elution with zero(ppm) water, 0.1 ~ 1mol/L NaCl; Follow the tracks of the effusive component of detection with the phenol sulfuric acid process, collect elution peak respectively, obtain elutriant;
(8) with step (7) gained elutriant; The Sephacryl S-400 HR gel-filtration column that overbalance is good carries out further separation and purification, carries out wash-out with 0.1mol/L NaCl, follows the tracks of with the phenol sulfuric acid process and detects effusive component; Collect elution peak respectively, obtain elutriant;
(9) elutriant in the step (8) is concentrated through the hollow fiber ultrafiltration membrane desalination and with it respectively, its molecular weight cut-off is 5000Dal (WP is controlled at 0.2MPa ~ 0.7Mpa, 20 ~ 60 ℃ of temperature), obtains golden mandarin orange polysaccharide soln;
(10) step (9) gained gold mandarin orange polysaccharide is concentrated into 3/10 ~ 2/5 of original volume, adds 80% ethanol alcohol precipitation and spend the night, after the lyophilize of gained deposition, pulverized 60 mesh sieves, promptly obtain fine gold mandarin orange polysaccharide dry powder.
Embodiment 2
(1) the fresh golden mandarin orange of screening is through cleaning, stoning, squeeze the juice, subsequent use;
(2) with golden mandarin orange juice by an endfeed mouth by the liquid material than the quantitative microwave extraction chamber of sending into of 10:1, microwave power microwave power 500W, microwave treatment time 18min;
(3) at the UW digester roIl, solvent is injected through under meter control amount by the other end fluid inlet, and solvent flows to discharge end from the liquid feeding end; When treating that flow of solvent is stablized, material helical feed in the step (2) is gone into the UW digester roIl, the liquid material is than being 30:1; Ultrasonic power 200W, UW 30min action time is under the promotion of screw; Solvent and the formation of golden mandarin orange powder flow to opposite continuous dynamic countercurrent and carry out the thorough mixing extraction, extract the gained vat liquor and get into next step operation by discharge port;
(4) the centrifugal insoluble impurities of removing of vat liquor in the step (3) obtains golden mandarin orange water extract;
(5) with gained gold mandarin orange water extract in the step (4), add 40 ℃ of enzymolysis 60min of proteolytic enzyme, adopt Sevag method deproteinated then, centrifugal behind the deproteinated 3 times, obtain supernatant;
(6) with step (5) gained supernatant, to cross macroporous adsorbent resin and decolour, selected macroporous adsorbent resin is D101, obtains golden mandarin orange polysaccharide soln after the decolouring;
(7) with step (6) gained gold mandarin orange polysaccharide soln; The DEAE Sepharose CL-6B ion exchange column that overbalance is good carries out separation and purification; Carry out stepwise elution with zero(ppm) water, 0.1 ~ 1mol/L NaCl; Follow the tracks of the effusive component of detection with the phenol sulfuric acid process, collect elution peak respectively, obtain elutriant;
(8) with step (7) gained elutriant; The Sephacryl S-400 HR gel-filtration column that overbalance is good carries out further separation and purification, carries out wash-out with 0.1mol/L NaCl, follows the tracks of with the phenol sulfuric acid process and detects effusive component; Collect elution peak respectively, obtain elutriant;
(9) elutriant in the step (8) is concentrated through the hollow fiber ultrafiltration membrane desalination and with it respectively, its molecular weight cut-off is 5000Dal (WP is controlled at 0.2MPa ~ 0.7Mpa, 20 ~ 60 ℃ of temperature), obtains golden mandarin orange polysaccharide soln;
(10) step (9) gained gold mandarin orange polysaccharide is concentrated into 3/10 ~ 2/5 of original volume, adds 80% ethanol alcohol precipitation and spend the night, after the lyophilize of gained deposition, pulverized 60 mesh sieves, promptly obtain fine gold mandarin orange polysaccharide dry powder.
Embodiment 3
(1) the fresh golden mandarin orange of screening is through cleaning, stoning, squeeze the juice, subsequent use;
(2) with golden mandarin orange dry powder by an endfeed mouth by the liquid material than the quantitative microwave extraction chamber of sending into of 12:1, microwave power microwave power 600W, microwave treatment time 15min;
(3) at the UW digester roIl, solvent is injected through under meter control amount by the other end fluid inlet, and solvent flows to discharge end from the liquid feeding end; When treating that flow of solvent is stablized, material helical feed in the step (2) is gone into the UW digester roIl, the liquid material is than being 28:1; Ultrasonic power 250W, UW 40min action time is under the promotion of screw; Solvent and the formation of golden mandarin orange powder flow to opposite continuous dynamic countercurrent and carry out the thorough mixing extraction, extract the gained vat liquor and get into next step operation by discharge port;
(4) the centrifugal insoluble impurities of removing of vat liquor in the step (3) obtains golden mandarin orange water extract;
(5) with gained gold mandarin orange water extract in the step (4), add 40 ℃ of enzymolysis 70min of proteolytic enzyme, adopt Sevag method deproteinated then, centrifugal behind the deproteinated 3 times, obtain supernatant;
(6) with step (5) gained supernatant, to cross macroporous adsorbent resin and decolour, selected macroporous adsorbent resin is D101, obtains golden mandarin orange polysaccharide soln after the decolouring;
(7) with step (6) gained gold mandarin orange polysaccharide soln; The DEAE Sepharose CL-6B ion exchange column that overbalance is good carries out separation and purification; Carry out stepwise elution with zero(ppm) water, 0.1 ~ 1mol/L NaCl; Follow the tracks of the effusive component of detection with the phenol sulfuric acid process, collect elution peak respectively, obtain elutriant;
(8) with step (7) gained elutriant; The Sephacryl S-400 HR gel-filtration column that overbalance is good carries out further separation and purification, carries out wash-out with 0.1mol/L NaCl, follows the tracks of with the phenol sulfuric acid process and detects effusive component; Collect elution peak respectively, obtain elutriant;
(9) elutriant in the step (8) is concentrated through the hollow fiber ultrafiltration membrane desalination and with it respectively, its molecular weight cut-off is 5000Dal (WP is controlled at 0.2MPa ~ 0.7Mpa, 20 ~ 60 ℃ of temperature), obtains golden mandarin orange polysaccharide soln;
(10) step (9) gained gold mandarin orange polysaccharide is concentrated into 3/10 ~ 2/5 of original volume, adds 80% ethanol alcohol precipitation and spend the night, after the lyophilize of gained deposition, pulverized 60 mesh sieves, promptly obtain fine gold mandarin orange polysaccharide dry powder.
The above is merely preferred embodiment of the present invention, and all equalizations of doing according to claim of the present invention change and modify, and all should belong to covering scope of the present invention.
Claims (3)
1. the method for efficiently extracting and purifying of a golden mandarin orange polysaccharide; It is characterized in that: fresh golden mandarin orange is squeezed the juice; Adopt microwave, UW Coupled Dynamic adverse current microwave leaching technology to carry out lixiviate; Obtain golden mandarin orange water extract through centrifuging; Adopt enzyme process to combine Sevag method deproteinated, macroporous adsorbent resin depigmentation to obtain golden mandarin orange polysaccharide soln again, separate, make the refining dry powder of golden mandarin orange polysaccharide again through ultrafiltration, concentrating under reduced pressure, alcohol precipitation, lyophilize, pulverizing through DEAE Sepharose CL-6B ion exchange column, Sephacryl S-400 HR gel column.
2. the method for efficiently extracting and purifying of golden mandarin orange polysaccharide according to claim 1 is characterized in that: the concrete steps of said method are:
(1) the fresh golden mandarin orange of screening is through cleaning, stoning, squeeze the juice, subsequent use;
(2) golden mandarin orange juice is quantitatively sent into microwave extraction chamber by the liquid material than 10:1 ~ 30:1 by an endfeed mouth, microwave power 400 ~ 620W, microwave treatment time 15 ~ 30min;
(3) at the UW digester roIl, solvent is injected through under meter control amount by the other end fluid inlet, and solvent flows to discharge port from fluid inlet; When treating that flow of solvent is stablized; Material helical feed in the step (2) is gone into the UW digester roIl, and the liquid material is than being 20:1 ~ 30:1, ultrasonic power 160 ~ 250W; UW 20 ~ 45min action time; Under the promotion of screw, solvent and the formation of golden mandarin orange flow to opposite continuous dynamic countercurrent and carry out the thorough mixing extraction, extract the gained vat liquor and get into next step operation by discharge port;
(4) the centrifugal insoluble impurities of removing of the vat liquor of step (3) obtains golden mandarin orange water extract;
(5) with the golden mandarin orange water extract of step (4), add 40 ℃ of enzymolysis 60 ~ 80min of proteolytic enzyme, adopt Sevag method deproteinated then, centrifugal behind the deproteinated 3 times, obtain supernatant;
(6) with the supernatant of step (5), to cross macroporous adsorbent resin and decolour, selected macroporous adsorbent resin is D101, obtains golden mandarin orange polysaccharide soln after the decolouring;
(7) with the golden mandarin orange polysaccharide soln of step (6); The DEAE Sepharose CL-6B ion exchange column that overbalance is good carries out separation and purification; NaCl solution with zero(ppm) water and 0.1 ~ 1mol/L carries out stepwise elution; Follow the tracks of the effusive component of detection with the phenol sulfuric acid process, collect elution peak respectively, obtain elutriant;
(8) with the elutriant of step (7); The Sephacryl S-400 HR gel-filtration column that overbalance is good carries out further separation and purification, carries out wash-out with the NaCl solution of 0.1 ~ 1mol/L, follows the tracks of with the phenol sulfuric acid process and detects effusive component; Collect elution peak respectively, obtain elutriant;
(9) elutriant with step (8) concentrates through the hollow fiber ultrafiltration membrane desalination and with it respectively, and its molecular weight cut-off is 5000Dal, and WP is controlled at 0.2MPa ~ 0.7MPa, and 20 ~ 60 ℃ of temperature obtain golden mandarin orange polysaccharide soln;
(10) step (9) gained gold mandarin orange polysaccharide is concentrated into 3/10 ~ 2/5 of original volume, adds massfraction and be 80% ethanolic soln alcohol precipitation and spend the night, after the lyophilize of gained deposition, pulverized 60 mesh sieves, promptly obtain the refining dry powder of golden mandarin orange polysaccharide.
3. the method for efficiently extracting and purifying of golden mandarin orange polysaccharide according to claim 2 is characterized in that: the solvent described in the step (3) is a pure water.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103005139A (en) * | 2013-01-06 | 2013-04-03 | 青岛亚博生物科技有限公司 | Preparation method of extract from fish tissues |
| CN103965369A (en) * | 2014-05-20 | 2014-08-06 | 河南中烟工业有限责任公司 | Ganoderan, extraction and purification method and application thereof as tobacco humectant |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100229948B1 (en) * | 1997-10-22 | 1999-12-01 | 최병대 | Extraction and Purification of Polysaccharide Sulfate from Off-White Peel |
| CN101434660A (en) * | 2008-08-01 | 2009-05-20 | 湖北省农业科学院农产品加工与核农技术研究所 | Method for extracting Ganoderma lucidum polysaccharide by ultrasonic wave-microwave combination |
| CN102295708A (en) * | 2011-06-17 | 2011-12-28 | 福建农林大学 | Optimization method for processing technology of active polysaccharide product |
| CN102558381A (en) * | 2012-02-16 | 2012-07-11 | 上海理工大学 | Method for extracting water-soluble soybean polysaccharide from low-temperature bean pulp through microwave-ultrasonic synergism |
-
2012
- 2012-08-27 CN CN201210307338.2A patent/CN102827299B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100229948B1 (en) * | 1997-10-22 | 1999-12-01 | 최병대 | Extraction and Purification of Polysaccharide Sulfate from Off-White Peel |
| CN101434660A (en) * | 2008-08-01 | 2009-05-20 | 湖北省农业科学院农产品加工与核农技术研究所 | Method for extracting Ganoderma lucidum polysaccharide by ultrasonic wave-microwave combination |
| CN102295708A (en) * | 2011-06-17 | 2011-12-28 | 福建农林大学 | Optimization method for processing technology of active polysaccharide product |
| CN102558381A (en) * | 2012-02-16 | 2012-07-11 | 上海理工大学 | Method for extracting water-soluble soybean polysaccharide from low-temperature bean pulp through microwave-ultrasonic synergism |
Non-Patent Citations (6)
| Title |
|---|
| 《International Journal of Biological Macromolecules》 20100210 Yiyong Chen et al. "Optimizationofultrasonic/microwaveassistedextraction(UMAE)of 第429-435页 1-3 第46卷, * |
| 《农业机械学报》 20100725 朱德文等 "茶叶微波超声波耦合动态逆流浸提工艺" 第136-140页 1-3 第41卷, 第7期 * |
| 《热带作物学报》 20120125 张怡等 "金柑多糖酶法脱蛋白工艺的研究" 第166-170页 1-3 第33卷, 第1期 * |
| YIYONG CHEN ET AL.: ""Optimizationofultrasonic/microwaveassistedextraction(UMAE)of", 《INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES》, vol. 46, 10 February 2010 (2010-02-10), pages 429 - 435 * |
| 张怡等: ""金柑多糖酶法脱蛋白工艺的研究"", 《热带作物学报》, vol. 33, no. 1, 25 January 2012 (2012-01-25), pages 166 - 170 * |
| 朱德文等: ""茶叶微波超声波耦合动态逆流浸提工艺"", 《农业机械学报》, vol. 41, no. 7, 25 July 2010 (2010-07-25), pages 136 - 140 * |
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