CN107540754A - The method for extraction and purification of oat bran active polysaccharide - Google Patents
The method for extraction and purification of oat bran active polysaccharide Download PDFInfo
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- CN107540754A CN107540754A CN201710737391.9A CN201710737391A CN107540754A CN 107540754 A CN107540754 A CN 107540754A CN 201710737391 A CN201710737391 A CN 201710737391A CN 107540754 A CN107540754 A CN 107540754A
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Abstract
The present invention's belongs to oat bran field of deep, more particularly to a kind of method for extraction and purification of oat bran active polysaccharide, its step include：Pretreatment, degreasing, ultrasonic-microwave coordinate extraction Polyose extraction, and finally yield, purity, monose composition, the anti-oxidant and hypolipidemic activity of the oat bran active polysaccharide of extraction are analyzed.The method for extraction and purification of oat bran active polysaccharide of the present invention, it is to utilize ultrasonic-microwave collaboration method extraction oat bran active polysaccharide mixture, having for being obtained again by dialysing, being concentrated under reduced pressure, being freeze-dried is anti-oxidant, lipid-loweringing regulation active polysaccharide, its extracting method is simple, operating cost is low, extraction time is short, and the oat bran active polysaccharide yield and purity of gained are respectively 8.45%, 93.2%, and with preferably anti-oxidant, reducing blood lipid regulation activity.
Description
Technical field
The invention belongs to oat bran deep process technology field, more particularly to a kind of extraction of oat bran active polysaccharide are pure
Change method.
Background technology
The outer tissue of oat that oat bran is the oat of cleaning or oatmeal obtains after milling, sieving.But swallow
Wheat springy texture, it is impossible to accomplish that wheat bran is completely separable with endosperm, cause to contain more endosperm silty in oat bran, with other
Cereal bran is compared, and substantial amounts of water-soluble dietary fiber is rich in oat bran, and these dietary fibers are that important feature is more
Sugar.But its healthcare function also is not deeply developed out by oat bran in addition to being used for bread, electuary etc. a little at present, pass through
Benefit of helping is relatively low, therefore, bioactive substance is extracted from oat bran, to increasing the economic value of oat bran and improving agriculture
The community income of byproduct is just particularly important.
Oat bran polyoses extract is a kind of multicomponent mixture, is primarily present in the granulated slag of oat bran non-starchy
In, its main component is beta glucan（62.3%）And pentosan（35.9%）And aldehydes matter etc., there is multiple biological activities work(
Can, it is good plant source functional polysaccharide.Oat polysaccharide after water extract-alcohol precipitation extracts is white, tasteless, neutral, has good
Good 26S Proteasome Structure and Function activity, wherein glutelin, phytic acid, heavy metal, agricultural chemicals and other microbial impurities content poles
Low, stability is preferable.Influence little with temperature and pH in terms of texture changes, and has retentiveness and network of fibers
Structure.So the necessary method to the extraction purification polysaccharide from oat bran is studied, its economic use value is improved.
The Study on extraction of oat bran polysaccharide is to develop the important step of oat bran, on its extraction process
Research has been achieved for certain achievement.Currently used extracting method includes Hot water extraction, alkaline extraction, sour formulation, enzyme formulation
Deng.But Hot water extraction is present the shortcomings of extraction time is long, yield is low, serious waste of resources；Regulation pH be present in alkalinity extraction
During produce the impurity such as a large amount of salt, be unfavorable for purifying and strong basicity composition may influence the functional activity of polysaccharide；Acid carries
Although method recovery rate is higher, beta glucan viscosity can be significantly reduced, a large amount of heat production of meeting, if applied to production, to consider to dissipate
Heat problem, practical application are more difficult；Enzyme formulation can effectively improve the yield of polysaccharide, and mild condition, make the biology of polysaccharide living
Property be not destroyed, but shortcoming be enzyme preparation cost height, be not easy to realize industrialized production.Therefore research invention is a kind of new effective
Extraction process prepares oat bran polysaccharide, and to overcome, the extraction time of original technology is longer, operating cost is high, complex process, carries
The defects of taking oat bran active polysaccharide relatively low is significant.
The content of the invention
A kind of it is contemplated that overcome the deficiencies in the prior art, there is provided extraction purification side of sharp oat bran active polysaccharide
Method, it is that oat bran is cleaned, concentrated, is freeze-dried and obtains that there is anti-oxidant, reducing blood lipid oat bran active polysaccharide
Preparation method, its extracting method is simple, and operating cost is low, and extraction time is short, obtained oat bran active polysaccharide have compared with
It is good it is anti-oxidant, fat biological is active.
The method for extraction and purification of oat bran active polysaccharide, step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 40~60 DEG C of drying, 30~80 mesh are ground to, obtain pre-processing oat
Wheat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 5~15 times of its weight, in the ethanol that purity is 85% by being immersed through a steps pretreatment oat bran, adds at 60~80 DEG C
Hot 1.5~3h of return stirring, it is cooled to room temperature and continues 10~18h of stirring, removes liquid, it is secondary 5~15 times of its weight of addition, pure
The ethanol for 85% is spent, 3~7h is stirred at room temperature, removes liquid, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin adds 2~8 times of its weight in step, purity is 99% acetone soln, stirring
After 30min, take and be deposited in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to 20~40 times of water of its weight, it is molten with sodium hydroxide solution or hydrochloric acid
Liquid adjusts pH to 8~12, under conditions of 200~600W of ultrasonic power, 400~800W of microwave power, 60~95 DEG C of temperature, extraction
5~20min of time, 2000~6000r/min centrifuging and taking supernatants are taken, precipitation continues to add water to original volume, as oat bran
Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 60~85 DEG C are heated to, adjust pH to 5.8~7.0, add bottom
Thing concentration is 0.3~1.5% heat-resistant alpha-amylase, stirs 10~35min, and desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extraction liquid obtained by step, 10~16 times of distilled water of its volume is added, then added
Enter the Sevage reagents that its volume accounting is 1/8~1/2, the min of stirring at normal temperature 15~30, take supernatant to add ethanol solution, make second
The volume fraction of alcohol is not less than 70%, stands 9~16 h at 1 DEG C~8 DEG C, takes precipitation, will precipitate respectively in 20~40 times of its volume
Absolute ethyl alcohol, wash successively in 20~40 times of anhydrous propanone and 20~40 times of diethyl ether solution, reuse 0.45 μm and receive
Filter membrane is filtered, and collects sediment, and deproteinized oat bran polysaccharide is made；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide obtained by step adds the ultra-pure water of 6~15 times of its weight, with 3500~7000Da
Dialysis membrane is dialysed 1~2 day using distilled water, during which changes water every 4~8h, the dialysis for taking its molecular weight to be more than 3500~7000Da
Liquid, oat bran polysaccharide extraction liquid is made；
（4）It is concentrated under reduced pressure：
Will（3）Oat bran polysaccharide extraction liquid is made in step and is concentrated into original volume under conditions of 40~70 DEG C, 0.09MPa
1/6~1/2, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）It is more to be freeze-dried obtained oat bran activity at -150~-80 DEG C for middle oat bran active polysaccharide concentrate
Sugar.
Further optimize the method for extraction and purification of oat bran active polysaccharide, its step is as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 45~50 DEG C of drying, 40~60 mesh are ground to, obtain pre-processing oat
Wheat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 10~15 times of its weight, in the ethanol that purity is 85% by being immersed through a steps pretreatment oat bran, at 60~80 DEG C
1.5~3h of stirring is heated to reflux, room temperature is cooled to and continues 10~18h of stirring, removes liquid, secondary 5~15 times of its weight of addition,
Purity is 85% ethanol, and 3~5h is stirred at room temperature, and removes liquid, and just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin adds 4~8 times of its weight in step, purity is 99% acetone soln, stirring
After 30min, take and be deposited in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to 20~40 times of water of its weight, it is molten with sodium hydroxide solution or hydrochloric acid
Liquid adjusts pH to 8~12, under conditions of 200~600W of ultrasonic power, 400~800W of microwave power, 60~95 DEG C of temperature, extraction
5~20min of time, 4000~6000r/min centrifuging and taking supernatants are taken, precipitation continues to add water to original volume, and oat bran is made
Thick many candies extract solution；
D, oat bran Thick many candies purify：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 60~85 DEG C are heated to, adjust pH to 5.8~7.0, add bottom
Thing concentration is 0.3~1.5% heat-resistant alpha-amylase, stirs 10~35min, and desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extraction liquid obtained by step, 10~16 times of distilled water of its volume is added, then
The Sevage reagents that its volume accounting is 1/4~1/2 are added, the min of stirring at normal temperature 15~30, takes supernatant to add ethanol solution, makes
The volume fraction of ethanol is not less than 70%, stands 9~12 h at 1 DEG C~8 DEG C, takes precipitation, will precipitate respectively in its volume 20~40
Washed successively in absolute ethyl alcohol again, 20~40 times of anhydrous propanone and 20~40 times of diethyl ether solution, reuse 0.45 μm
NF membrane is filtered, and collects sediment, and deproteinized oat bran polysaccharide is made；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide obtained by step, the ultra-pure water of 6~15 times of its weight is added, with 3500~
7000Da dialysis membranes are dialysed 1~2 day using distilled water, are during which changed water every 4~6h, are taken its molecular weight to be more than 3500~7000Da
Dialyzate, oat bran polysaccharide extraction liquid is made；
（4）It is concentrated under reduced pressure：
Will（3）Oat bran polysaccharide extraction liquid is made in step and is concentrated into original volume under conditions of 40~70 DEG C, 0.09MPa
1/6~1/2, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）It is more to be freeze-dried obtained oat bran activity at -150~-80 DEG C for middle oat bran active polysaccharide concentrate
Sugar.
As the further optimization of the present invention, the De-fatted oat bran tare weight in step c is extracted 2~4 times again, merges extraction
Liquid, oat bran Thick many candies extract solution is made.
As the further optimization of the present invention, step d（5）In, oat bran active polysaccharide concentrate is at -30~-15 DEG C
After the h of pre-freeze 24~36, then it is freeze-dried, produces oat bran active polysaccharide.
The method for extraction and purification of oat bran active polysaccharide of the present invention, it is to utilize ultrasonic-microwave collaboration method extraction oat bran
Active polysaccharide mixture, then what is obtained by dialysing, being concentrated under reduced pressure, being freeze-dried have anti-oxidant, lipid-loweringing regulation active polysaccharide,
Its extracting method is simple, and operating cost is low, and extraction time is short, and the oat bran active polysaccharide yield and purity of gained are respectively
8.45%th, 93.2%, and with preferably anti-oxidant, reducing blood lipid regulation activity.
Brief description of the drawings
Fig. 1 be various concentrations oat bran polyoses extract and Vc to DPPH clearance rate；
Fig. 2 be various concentrations oat bran polyoses extract and Vc to O2-Clearance rate；
Fig. 3 is the monose of oat bran active polysaccharide；
Fig. 4 is influence of the oat bran active polysaccharide to high fat lipid of mice six.
Embodiment
Embodiment 1
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 40 DEG C of drying, 30 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
The g of oat bran 70 will be pre-processed through a steps, and add the mL of 85 % 350 ethanol, stirring 1.5 is heated to reflux at 60 DEG C
H, it is cooled to room temperature and continues to stir 10 h, remove liquid, add the mL of 85 % 350 ethanol, 3 h are stirred at room temperature, remove liquid,
Just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin, adds 99 % 150mL acetone soln, after stirring 30min, it is heavy to take in step
Form sediment in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
By the g of De-fatted oat bran skin 50 made from b step, 1000mL water is added, is adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 8, under conditions of the W of ultrasonic power 200, the W of microwave power 400,60 DEG C of temperature, extraction time 5 min, 2000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The mL of oat bran Thick many candies extract solution 500 that step c is obtained, 60 DEG C are heated to, adjust pH to 5.8, add substrate
Concentration is 0.3 % heat-resistant alpha-amylase, stirs 10 min, and desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）The mL of desizing oat bran polysaccharide extraction liquid 500 obtained by step, adds 5000 mL distilled water, adds
650 mL Sevage reagents, the min of stirring at normal temperature 15, take supernatant add ethanol solution, and make ethanol volume fraction be 70%,
9 h are stood at 1 DEG C, take precipitation, by the precipitation absolute ethyl alcohol in 100 mL, 100 mL anhydrous propanone and 100 mL respectively
Washed successively in diethyl ether solution, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized oat bran is made
Polysaccharide；
（3）Go low molecular weight impurities：
Will（2）The mL of deproteinized oat bran polysaccharide 100 obtained by step, 600 mL ultra-pure water is added, dialysed with 3500 Da
Film is dialysed 1 day using distilled water, during which changes water once every 4 h, the dialyzate for taking its molecular weight to be more than 3500 Da, swallow is made
Wheat bran skin polysaccharide extraction liquid；
（4）It is concentrated under reduced pressure：
Will（3）The middle mL of oat bran polysaccharide extraction liquid 500 after dialysis, it is concentrated under conditions of 40 DEG C, 0.09MPa
80 mL, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）The middle mL of oat bran active polysaccharide concentrate 80, after -30 DEG C of h of pre-freeze 24, is freeze-dried at -80 DEG C
Oat bran active polysaccharide is made；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 2
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 60 DEG C of drying, 80 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
The g of oat bran 70 will be pre-processed through a steps, and add the mL of 85 % 1050 ethanol, stirring 3 is heated to reflux at 80 DEG C
H, it is cooled to room temperature and continues to stir 18 h, remove liquid, add the mL of 85 % 1050 ethanol, 7 h are stirred at room temperature, remove liquid
Body, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin, adds 99 % 560mL acetone soln, after stirring 30min, it is heavy to take in step
Form sediment in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
By the g of De-fatted oat bran skin 50 made from b step, 2800mL water is added, is adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 12, under conditions of the W of ultrasonic power 600, the W of microwave power 800,95 DEG C of temperature, extraction time 20 min, 6000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The mL of oat bran Thick many candies extract solution 500 that step c is obtained, 85 DEG C are heated to, adjust pH to 7.0, add substrate
Concentration is 1.5 % heat-resistant alpha-amylase, stirs 35 min, and desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）The mL of desizing oat bran polysaccharide extraction liquid 500 obtained by step, adds 8000 mL distilled water, adds
4000 mL Sevage reagents, stirring at normal temperature 30min, supernatant is taken to add ethanol solution, the volume fraction for making ethanol is 70%,
8 DEG C of 16 h of standing, take the diethyl ether solution of the precipitation 5g absolute ethyl alcohol in 200 mL, 200 mL anhydrous propanone and 200 mL respectively
In wash successively, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized oat bran polysaccharide is made；
（3）Go low molecular weight impurities：
Will（2）The mL of deproteinized oat bran polysaccharide 100 obtained by step, 1500 mL ultra-pure water is added, dialysed with 7000 Da
Film is dialysed 2 days using distilled water, during which changes water once every 8h, the dialyzate for taking its molecular weight to be more than 7000 Da, swallow is made
Wheat bran skin polysaccharide extraction liquid；
（4）It is concentrated under reduced pressure：
Will（3）The middle mL of oat bran polysaccharide extraction liquid 500 after dialysis, subtracted under conditions of 70 DEG C, 0.09MPa
Pressure is concentrated into 250 mL, and oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）The middle mL of oat bran active polysaccharide concentrate 80, after -30 DEG C of pre-freeze 36h, is freeze-dried at -150 DEG C
Oat bran active polysaccharide is made；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 3
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 45 DEG C of drying, 40 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
The g of oat bran 70 will be pre-processed through a steps, and add the mL of 85 % 700 ethanol, stirring 1.5 is heated to reflux at 60 DEG C
H, it is cooled to room temperature and continues to stir 10 h, remove liquid, add the mL of 85 % 350 ethanol, 3 h are stirred at room temperature, remove liquid,
Just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin, adds 99 % 280mL acetone soln, after stirring 30min, it is heavy to take in step
Form sediment in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
By the g of De-fatted oat bran skin 50 made from b step, 1000mL water is added, is adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 8, under conditions of the W of ultrasonic power 200, the W of microwave power 400,60 DEG C of temperature, extraction time 5 min, 4000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The mL of oat bran Thick many candies extract solution 500 that step c is obtained, 60 DEG C are heated to, adjust pH to 5.8, add substrate
Concentration is 0.3 % heat-resistant alpha-amylase, stirs 10 min, and desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）The mL of desizing oat bran polysaccharide extraction liquid 500 obtained by step, adds 5000 mL distilled water, adds
1250 mL Sevage reagents, stirring at normal temperature 15min, take supernatant to add the ethanol solution of certain volume, make the volume integral of ethanol
Number is 70%, and 9h, taking precipitate the 5g absolute ethyl alcohol in 100 mL, 100 mL anhydrous propanone and 100 mL respectively are stood at 1 DEG C
Diethyl ether solution in wash successively, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized oat bran is made
Skin polysaccharide；
（3）Go low molecular weight impurities：
Will（2）The mL of deproteinized oat bran polysaccharide 100 obtained by step, 600 mL ultra-pure water is added, dialysed with 3500 Da
Film is dialysed 1 day using distilled water, during which changes water once every 4h, the dialyzate for taking its molecular weight to be more than 3500 Da, swallow is made
Wheat bran skin polysaccharide extraction liquid；
（4）It is concentrated under reduced pressure：
Will（3）The mL of oat bran polysaccharide extraction liquid 500 in step after dialysis, carry out depressurizing under conditions of 40 DEG C dense
80 mL are reduced to, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）The mL of oat bran active polysaccharide concentrate 80 in step, after -15 DEG C of pre-freeze 24h, is freezed at -150 DEG C
Dry obtained oat bran active polysaccharide；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 4
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 50 DEG C of drying, 60 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
The g of oat bran 70 will be pre-processed through a steps, and add the mL of 85 % 1050 ethanol, stirring 3 is heated to reflux at 80 DEG C
H, it is cooled to room temperature and continues to stir 18 h, remove liquid, add the mL of 85 % 1050 ethanol, 5 h are stirred at room temperature, remove liquid
Body, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin, adds 99 % 560mL acetone soln, after stirring 30min, it is heavy to take in step
Form sediment in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
By the g of De-fatted oat bran skin 50 made from b step, 2000mL water is added, is adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 12, under conditions of the W of ultrasonic power 600, the W of microwave power 800,95 DEG C of temperature, extraction time 20 min, 6000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The mL of oat bran Thick many candies extract solution 500 that step c is obtained, 85 DEG C are heated to, adjust pH to 7.0, add substrate
Concentration is 1.5 % heat-resistant alpha-amylase, stirs 35 min, and desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）The mL of desizing oat bran polysaccharide extraction liquid 500 obtained by step, adds 8000 mL distilled water, adds
2000 mL Sevage reagents, stirring at normal temperature 30min, supernatant is taken to add ethanol solution, the volume fraction for making ethanol is 70%,
8 DEG C of 12 h of standing, take the diethyl ether solution of the precipitation 5g absolute ethyl alcohol in 200 mL, 200 mL anhydrous propanone and 200 mL respectively
In wash successively, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized oat bran polysaccharide is made；
（3）Go low molecular weight impurities：
Will（2）The mL of deproteinized oat bran polysaccharide 100 obtained by step, 1500 mL ultra-pure water is added, dialysed with 7000 Da
Film is dialysed 2 days using distilled water, during which changes water once every 6h, the dialyzate for taking its molecular weight to be more than 7000 Da, swallow is made
Wheat bran skin polysaccharide extraction liquid；
（4）It is concentrated under reduced pressure：
Will（3）The middle mL of oat bran polysaccharide extraction liquid 500 after dialysis, subtracted under conditions of 70 DEG C, 0.09MPa
Pressure is concentrated into 250 mL, and oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）The middle mL of oat bran active polysaccharide concentrate 80, after -15 DEG C of pre-freeze 36h, is freeze-dried at -150 DEG C
Oat bran active polysaccharide is made；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 5
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 48 DEG C of drying, 50 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
The g of oat bran 70 will be pre-processed through a steps, and add the mL of 85 % 840 ethanol, and be heated to reflux stirring 2 h at 80 DEG C,
It is cooled to room temperature to continue to stir 14 h, removes liquid, add the mL of 85 % 700 ethanol, 4 h are stirred at room temperature, removes liquid, system
Obtain just De-fatted oat bran skin；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin, adds 99 % 420mL acetone soln, after stirring 30min, it is heavy to take in step
Form sediment, in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
By the g of De-fatted oat bran skin 50 made from b step, 1500mL water is added, is adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 10, under conditions of the W of ultrasonic power 400, the W of microwave power 600,80 DEG C of temperature, extraction time 15 min, 5000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The mL of oat bran Thick many candies extract solution 500 that step c is obtained, 70 DEG C are heated to, adjust pH to 6.5, add substrate
Concentration is 0.9 % heat-resistant alpha-amylase, stirs 25 min, and desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）The mL of desizing oat bran polysaccharide extraction liquid 500 obtained by step, adds 4500 mL distilled water, adds
1800 mL Sevage reagents, stirring at normal temperature 30min, supernatant is taken to add ethanol solution, the volume fraction for making ethanol is 70%,
5 DEG C of 10 h of standing, take the diethyl ether solution of the precipitation 5g absolute ethyl alcohol in 150 mL, 150 mL anhydrous propanone and 150 mL respectively
In wash successively, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized oat bran polysaccharide is made；
（3）Go low molecular weight impurities：
Will（2）The mL of deproteinized oat bran polysaccharide 100 obtained by step, 1000 mL ultra-pure water is added, dialysed with 5000 Da
Film is dialysed 2 days using distilled water, during which changes water once every 5h, the dialyzate for taking its molecular weight to be more than 5000 Da, swallow is made
Wheat bran skin polysaccharide extraction liquid；
（4）It is concentrated under reduced pressure：
Will（3）The middle mL of oat bran polysaccharide extraction liquid 500 after dialysis, subtracted under conditions of 55 DEG C, 0.09MPa
Pressure is concentrated into 200 mL, and oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）The middle mL of oat bran active polysaccharide concentrate 80, after -20 DEG C of pre-freeze 30h, is freeze-dried at -150 DEG C
Oat bran active polysaccharide is made；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 6
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 40 DEG C of drying, 30 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 5 times of its weight, in the ethanol that purity is 85 % by being immersed through a steps pretreatment oat bran, is heated to reflux at 60 DEG C
1.5 h are stirred, room temperature is cooled to and continues to stir 10 h, remove liquid, secondary 5 times of its weight of addition, the ethanol that purity is 85 %
In, 3 h are stirred at room temperature, remove liquid, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin in step, adds 2 times of its weight, the acetone soln that purity is 99 %, stirring
After 30min, take and be deposited in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to the water of 20 times of its weight, adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 8, under conditions of the W of ultrasonic power 200, the W of microwave power 400,60 DEG C of temperature, extraction time 5 min, 2000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 60 DEG C are heated to, adjust pH to 5.8, adding concentration of substrate is
0.3 % heat-resistant alpha-amylase, 10 min are stirred, desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extraction liquid obtained by step, adds 10 times of distilled water of its volume, adds it
Volume accounting is 1/8 Sevage reagents, the min of stirring at normal temperature 15, takes supernatant to add ethanol solution, and make the volume integral of ethanol
Number is 70%, stands 9 h at 1 DEG C, takes precipitation, by the precipitation absolute ethyl alcohol in 20 times of its volume, 20 times of anhydrous propanone respectively
Washed successively with 20 times of diethyl ether solution, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized is made
Oat bran polysaccharide；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide obtained by step, the ultra-pure water of 6 times of its volume is added, is adopted with 3500 Da dialysis membranes
Dialysed 1 day with distilled water, during which change water once every 4 h, the dialyzate for taking its molecular weight to be more than 3500 Da, oat bran is made
Skin polysaccharide extraction liquid；
（4）It is concentrated under reduced pressure：
Will（3）The middle oat bran polysaccharide extraction liquid after dialysis, original volume is concentrated under conditions of 40 DEG C, 0.09MPa
1/6, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）Middle oat bran active polysaccharide concentrate, after -30 DEG C of h of pre-freeze 24, it is freeze-dried at -80 DEG C and swallow is made
Wheat bran skin active polysaccharide；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 7
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 60 DEG C of drying, 80 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 15 times of its weight, in the ethanol that purity is 85 % by being immersed through a steps pretreatment oat bran, is heated back at 80 DEG C
Stream 3 h of stirring, it is cooled to room temperature and continues to stir 18 h, removes liquid, secondary 15 times of its weight of addition, the ethanol that purity is 85 %
In, 7 h are stirred at room temperature, remove liquid, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin in step, adds 8 times of its weight, the acetone soln that purity is 99 %, stirring
After 30min, take and be deposited in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to the water of 16 times of its weight, adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 12, under conditions of the W of ultrasonic power 600, the W of microwave power 800,95 DEG C of temperature, extraction time 20 min, 6000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 85 DEG C are heated to, adjust pH to 7.0, adding concentration of substrate is
1.5 % heat-resistant alpha-amylase, 35 min are stirred, desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extraction liquid obtained by step, adds 16 times of distilled water of its volume, adds it
Volume accounting is 1/2 Sevage reagents, and stirring at normal temperature 30min takes supernatant to add ethanol solution, and the volume fraction for making ethanol is
70%, 16 h are stood at 8 DEG C, take precipitation, by the precipitation absolute ethyl alcohol in 40 times of its volume, 40 times of anhydrous propanone and 40 respectively
Washed successively in diethyl ether solution again, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized oat is made
Bran polysaccharide；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide obtained by step, the ultra-pure water of 15 times of its volume is added, with 7000 Da dialysis membranes
Dialysed 2 days using distilled water, during which change water once every 8h, the dialyzate for taking its molecular weight to be more than 7000 Da, oat is made
Bran polysaccharide extract solution；
（4）It is concentrated under reduced pressure：
Will（3）The middle oat bran polysaccharide extraction liquid after dialysis, it is concentrated under reduced pressure under conditions of 70 DEG C, 0.09MPa
To the 1/2 of original volume, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）Middle oat bran active polysaccharide concentrate, after -30 DEG C of pre-freeze 36h, it is freeze-dried at -150 DEG C and swallow is made
Wheat bran skin active polysaccharide；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 8
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 45 DEG C of drying, 40 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 10 times of its weight, in the ethanol that purity is 85 % by being immersed through a steps pretreatment oat bran, is heated back at 60 DEG C
Stream 1.5 h of stirring, it is cooled to room temperature and continues to stir 10 h, removes liquid, secondary 5 times of its weight of addition, the second that purity is 85 %
In alcohol, 3 h are stirred at room temperature, remove liquid, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin in step, adds 4 times of its weight, the acetone soln that purity is 99 %, stirring
After 30min, take and be deposited in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to the water of 20 times of its weight, adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 8, under conditions of the W of ultrasonic power 200, the W of microwave power 400,60 DEG C of temperature, extraction time 5 min, 4000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 60 DEG C are heated to, adjust pH to 5.8, adding concentration of substrate is
0.3 % heat-resistant alpha-amylase, 10 min are stirred, desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extraction liquid extract solution obtained by step, 10 times of distilled water of its volume is added, then
The Sevage reagents that its volume accounting is 1/4 are added, stirring at normal temperature 15min, takes supernatant to add the ethanol solution of certain volume, makes
The volume fraction of ethanol is 70%, and 9h is stood at 1 DEG C, will precipitate the absolute ethyl alcohol in 20 times of its volume, anhydrous the third of 20 times respectively
Washed successively in ketone and 20 times of diethyl ether solution, reuse 0.45 μm of NF membrane and filtered, collect sediment, be made and remove egg
White oat bran polysaccharide；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide obtained by step, the ultra-pure water of 6 times of its volume is added, is adopted with 3500 Da dialysis membranes
Dialysed 1 day with distilled water, during which change water once every 4h, the dialyzate for taking its molecular weight to be more than 3500 Da, oat bran is made
Skin polysaccharide extraction liquid；
（4）It is concentrated under reduced pressure：
Will（3）Oat bran polysaccharide extraction liquid in step after dialysis, carry out being concentrated under reduced pressure into original under conditions of 40 DEG C
The 1/6 of volume, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）Oat bran active polysaccharide concentrate in step, after -15 DEG C of pre-freeze 24h, system is freeze-dried at -150 DEG C
Obtain oat bran active polysaccharide；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 9
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 50 DEG C of drying, 60 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 15 times of its weight, in the ethanol that purity is 85 % by being immersed through a steps pretreatment oat bran, is heated back at 80 DEG C
Stream 3 h of stirring, it is cooled to room temperature and continues to stir 18 h, removes liquid, secondary 15 times of its weight of addition, the ethanol that purity is 85 %
In, 5 h are stirred at room temperature, remove liquid, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin in step, adds 8 times of its weight, the acetone soln that purity is 99 %, stirring
After 30min, take and be deposited in room temperature nature volatile dry 12h, obtain De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to the water of 40 times of its weight, adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 12, under conditions of the W of ultrasonic power 600, the W of microwave power 800,95 DEG C of temperature, extraction time 20 min, 6000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 85 DEG C are heated to, adjust pH to 7.0, adding concentration of substrate is
1.5 % heat-resistant alpha-amylase, 35 min are stirred, desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extract solution obtained by step, 16 times of distilled water of its volume is added, then added
Enter the Sevage reagents that its volume accounting is 1/4, stirring at normal temperature 30min, take supernatant to add ethanol solution, make the volume integral of ethanol
Number is 70%, stands 12 h at 8 DEG C, takes precipitation, by the precipitation absolute ethyl alcohol in 40 times of its volume, 40 times of anhydrous propanone respectively
Washed successively with 40 times of diethyl ether solution, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized is made
Oat bran polysaccharide；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide obtained by step, the ultra-pure water of 15 times of its volume is added, with 7000 Da dialysis membranes
Dialysed 2 days using distilled water, during which change water once every 6h, the dialyzate for taking its molecular weight to be more than 7000 Da, oat is made
Bran polysaccharide extract solution；
（4）It is concentrated under reduced pressure：
Will（3）The middle oat bran polysaccharide extraction liquid after dialysis, it is concentrated under reduced pressure under conditions of 70 DEG C, 0.09MPa
To the 1/2 of original volume, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）Middle oat bran active polysaccharide concentrate, after -15 DEG C of pre-freeze 36h, it is freeze-dried at -150 DEG C and swallow is made
Wheat bran skin active polysaccharide；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Embodiment 10
The method for extraction and purification of oat bran active polysaccharide, specific implementation step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 48 DEG C of drying, 50 mesh are ground to, obtain pre-processing oat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 12 times of its weight, in the ethanol that purity is 85 % by being immersed through a steps pretreatment oat bran, is heated back at 80 DEG C
Stream 2 h of stirring, it is cooled to room temperature and continues to stir 14 h, removes liquid, secondary 12 times of its weight of addition, the ethanol that purity is 85 %
In, 4 h are stirred at room temperature, remove liquid, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin in step, adds 6 times of its weight, the acetone soln that purity is 99 %, stirring
After 30min, precipitation is taken, in room temperature nature volatile dry 12h, obtains De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to the water of 30 times of its weight, adjusted with sodium hydroxide solution or hydrochloric acid solution
PH to 10, under conditions of the W of ultrasonic power 400, the W of microwave power 600,80 DEG C of temperature, extraction time 15 min, 5000 r/
Min centrifuging and taking supernatants, as oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 70 DEG C are heated to, adjust pH to 6.5, adding concentration of substrate is
0.9 % heat-resistant alpha-amylase, 25 min are stirred, desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extraction liquid obtained by step, adds 9 times of distilled water of its volume, adds its body
The Sevage reagents that product accounting is 1/3, stirring at normal temperature 30min take supernatant to add ethanol solution, and the volume fraction for making ethanol is
70%, 10 h are stood at 5 DEG C, take precipitation, by the precipitation absolute ethyl alcohol in 30 times of its volume, 30 times of anhydrous propanone and 30 respectively
Washed successively in diethyl ether solution again, reuse 0.45 μm of NF membrane and filtered, collect sediment, deproteinized oat is made
Bran polysaccharide；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide obtained by step, the ultra-pure water of 10 times of its volume is added, with 5000 Da dialysis membranes
Dialysed 2 days using distilled water, during which change water once every 5h, the dialyzate for taking its molecular weight to be more than 5000 Da, oat is made
Bran polysaccharide extract solution；
（4）It is concentrated under reduced pressure：
Will（3）The middle oat bran polysaccharide extraction liquid after dialysis, it is concentrated under reduced pressure under conditions of 55 DEG C, 0.09MPa
To the 2/5 of original volume, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）Middle oat bran active polysaccharide concentrate, after -20 DEG C of pre-freeze 30h, it is freeze-dried at -150 DEG C and swallow is made
Wheat bran skin active polysaccharide；
Wherein, Sevage is to be mixed by chloroform and n-butanol by 5: 1 volume ratio.
Formed using the monose of oat bran active polysaccharide made from the above method, as shown in table 1, oat bran activity is more
The monose composition of sugar is mainly glucose 61.3%, xylose 20.3%, arabinose 16.3%, additionally contains a small amount of galactolipin
1.6%th, rhamnose 0.3% and mannose 0.2%.
Using the oat bran polyoses extract and Vc of various concentrations made from the above method to DPPH clearance rate such as
Shown in Fig. 1, oat bran polysaccharide extraction thing gradually steps up with the increase of concentration, its DPPH clearance rate, and DPPH is clear
A certain amount effect relation is presented with concentration in removing solid capacity.Variance analysis shows the oat bran active polysaccharide of various concentrations to DPPH
Scavenging action there were significant differences.When oat bran active polysaccharide concentration is 2.5 mg/mL, it reaches to DPPH clearance rates
93.91%, now only than the DPPH clearance rates of Vc under same concentration（98.98%）Low 5.1%.
Using the oat bran polyoses extract and Vc of various concentrations made from the above method to O2-Clearance rate, such as scheme
Shown in 2, in the range of experimental concentration, the increase with oat bran active polysaccharide with concentration, it is to O2-Clearance rate constantly carry
It is high.When concentration is 1mg/mL, oat bran active polysaccharide is to O2-Clearance rate reached 58.31%, it is dense entirely testing
It is to O in the range of degree2-Clearance rate it is suitable with Vc, while oat bran active polysaccharide and VCEC50Respectively（1.09±
0.02）With（0.69±0.09）Mg/mL, it is also seen that both remove O2-Ability it is suitable, illustrate oat bran activity it is more
Sugar has very strong O2-Scavenging activity.
It is as shown in table 2 using influence of the oat bran active polysaccharide made from the above method to high fat lipid of mice six,
There is hyperlipemia symptom after high lipid food is fed in rat, and its serum TC, TG, HDL-C, LDL-C, apoB are changed significantly（P ＜
0.05）It is and relatively stable within follow-up experimental period.3rd ~ 12 groups：There is hyperlipemia after high lipid food is fed in rat
Symptom, its serum TC, TG, HDL-C, LDL-C, apoB are changed significantly（P ＜ 0.05）, after different preparation gavages, blood fat six
Level makes moderate progress, wherein most significantly the 4th group of TC improvements；Most significantly the 5th group of TG improvements；HDL-
Most significantly the 4th group of C improvements；Most significantly the 7th group of LDL-C improvements；ApoA improvements are most significant
It is the 5th group；Most significantly the tenth group of apoB improvements.Comprehensive comparison：Agent in 5th group of oat bran active polysaccharide
Amount group blood lipid level improvement is optimal.
Table 2 annotates as follows：1. blank control group（One）, high lipid food group（Two）, positive controls（Three）, oat bran live
Property polysaccharide high dose group（Four）, oat bran active polysaccharide middle dose group（Five）, oat bran active polysaccharide low dose group（Six）、
Oat bran powder high dose group（Seven）, oat bran powder middle dose group（Eight）, oat bran powder low dose group（Nine）, beta glucan
High dose group（Ten）, beta glucan middle dose group（11）, beta glucan low dose group（12）；2. others is rat serum in table
Clear TC, TG, HDL-C, LDL-C, apo-A, apo-B testing result twice（Twice respectively before gavage, after gavage）；It is 3. same
Lowercase difference significant difference on row（P ＜ 0.05）, capitalization difference difference is extremely notable（P ＜ 0.01）.
Claims (4)
1. the method for extraction and purification of oat bran active polysaccharide, step are as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 40~60 DEG C of drying, 30~80 mesh are ground to, obtain pre-processing oat
Wheat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 5~15 times of its weight, in the ethanol that purity is 85% by being immersed through a steps pretreatment oat bran, adds at 60~80 DEG C
Hot 1.5~3h of return stirring, it is cooled to room temperature and continues 10~18h of stirring, removes liquid, it is secondary 5~15 times of its weight of addition, pure
The ethanol for 85% is spent, 3~7h is stirred at room temperature, removes liquid, just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin adds 2~8 times of its weight in step, purity is 99% acetone soln, stirring
Liquid is removed after 30min, in room temperature nature volatile dry 12h, obtains De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to 20~40 times of water of its weight, it is molten with sodium hydroxide solution or hydrochloric acid
Liquid adjusts pH to 8~12, under conditions of 200~600W of ultrasonic power, 400~800W of microwave power, 60~95 DEG C of temperature, extraction
Take 5~20min of time, as 2000~6000r/min centrifuging and taking supernatants, oat bran Thick many candies extract solution；
D, oat bran Thick many candies extract solution purifies：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 60~85 DEG C are heated to, adjust pH to 5.8~7.0, Ran Houjia
Enter 0.3~1.5% heat-resistant alpha-amylase of oat bran Thick many candies extract solution quality, stir 10~35min, desizing is made
Oat bran polysaccharide extraction liquid；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extraction liquid obtained by step, 10~16 times of distilled water of its volume is added, then added
Enter the Sevage reagents that its volume accounting is 1/8~1/2, the min of stirring at normal temperature 15~30, take supernatant to add ethanol solution, make second
The volume fraction of alcohol be not less than 70%, 1 DEG C~8 DEG C stand 9~16 h, taking precipitate, by sediment respectively its volume 20~
Washed successively in 40 times of absolute ethyl alcohol, 20~40 times of anhydrous propanone and 20~40 times of diethyl ether solution, reuse 0.45 μm
NF membrane filtered, collect sediment, as deproteinized oat bran polysaccharide；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide obtained by step adds the ultra-pure water of 6~15 times of its weight, with 3500~7000Da
Dialysis membrane is dialysed 1~2 day using distilled water, during which changes water every 4~8h, the dialysis for taking its molecular weight to be more than 3500~7000Da
Liquid, oat bran polysaccharide extraction liquid is made；
（4）It is concentrated under reduced pressure：
Will（3）Oat bran polysaccharide extraction liquid is made in step and is concentrated into original volume under conditions of 40~70 DEG C, 0.9MPa
1/6~1/2, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）Oat bran active polysaccharide concentrate is made at -150~-80 DEG C in step, is freeze-dried and oat bran work is made
Property polysaccharide.
2. the method for extraction and purification of oat bran active polysaccharide according to claim 1, its operation further optimized step
It is rapid as follows：
A, pre-process：After oat bran is gone into the removal of impurity, 45~50 DEG C of drying, 40~60 mesh are ground to, obtain pre-processing oat
Wheat bran；
B, degreasing：
（1）Alcohol degreasing：
It is 10~15 times of its weight, in the ethanol that purity is 85% by being immersed through a steps pretreatment oat bran, at 60~80 DEG C
1.5~3h of stirring is heated to reflux, room temperature is cooled to and continues 10~18h of stirring, removes liquid, secondary 5~15 times of its weight of addition,
Purity is 85% ethanol, and 3~5h is stirred at room temperature, and removes liquid, and just De-fatted oat bran skin is made；
（2）Acetone degreasing：
Will（1）Obtained just De-fatted oat bran skin adds 4~8 times of its weight in step, purity is 99% acetone soln, stirring
Liquid is removed after 30min, in room temperature nature volatile dry 12h, obtains De-fatted oat bran skin；
C, Polyose extraction：
De-fatted oat bran skin made from b step is added to 20~40 times of water of its weight, it is molten with sodium hydroxide solution or hydrochloric acid
Liquid adjusts pH to 8~12, under conditions of 200~600W of ultrasonic power, 400~800W of microwave power, 60~95 DEG C of temperature, extraction
5~20min of time, 4000~6000r/min centrifuging and taking supernatants are taken, precipitation continues to add water to original volume, and oat bran is made
Thick many candies extract solution；
D, oat bran Thick many candies purify：
（1）Desizing：
The oat bran Thick many candies extract solution that step c is obtained, 60~85 DEG C are heated to, adjust pH to 5.8~7.0, add bottom
Thing concentration is 0.3~1.5% heat-resistant alpha-amylase, stirs 10~35min, and desizing oat bran polysaccharide extraction liquid is made；
（2）Deproteinized：
By d（1）Desizing oat bran polysaccharide extraction liquid obtained by step, 10~16 times of distilled water of its volume is added, then
The Sevage reagents that its volume accounting is 1/4~1/2 are added, the min of stirring at normal temperature 15~30, takes supernatant to add ethanol solution, makes
The volume fraction of ethanol be not less than 70%, 1 DEG C~8 DEG C stand 9~12 h, taking precipitate, will precipitation respectively its volume 20~
Washed successively in 40 times of absolute ethyl alcohol, 20~40 times of anhydrous propanone and 20~40 times of diethyl ether solution, reuse 0.45 μm
NF membrane filtered, collect sediment, deproteinized oat bran polysaccharide extraction liquid be made；
（3）Go low molecular weight impurities：
Will（2）Deproteinized oat bran polysaccharide extraction liquid obtained by step, the ultra-pure water of 6~15 times of its weight is added, with 3500
~7000Da dialysis membranes using distilled water dialyse 1~2 day, during which change water every 4~6h, take its molecular weight be more than 3500~
7000Da dialyzate, oat bran polysaccharide extraction liquid is made；
（4）It is concentrated under reduced pressure：
Will（3）Oat bran polysaccharide extraction liquid is made in step and is concentrated into original volume under conditions of 40~70 DEG C, 0.9MPa
1/6~1/2, oat bran active polysaccharide concentrate is made；
（5）Freeze-drying：
Will（4）Oat bran active polysaccharide concentrate is made at -150~-80 DEG C in step, is freeze-dried and oat bran work is made
Property polysaccharide.
3. the method for extraction and purification of oat bran active polysaccharide according to claim 1 or 2, it is characterised in that：By step c
In De-fatted oat bran tare weight extract again 2~4 times, merge extract solution, oat bran Thick many candies extract solution be made.
4. the method for extraction and purification of oat bran active polysaccharide according to claim 1 or 2, it is characterised in that：Step d
（5）In, oat bran active polysaccharide concentrate is after -30~-15 DEG C of h of pre-freeze 24~36, then is freeze-dried, and produces swallow
Wheat bran skin active polysaccharide.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108752491A (en) * | 2018-05-21 | 2018-11-06 | 黑龙江八农垦大学 | Ultrasound-microwave radiation technology water extraction extracts folium isatidis active polysaccharide technique |
| CN109134697A (en) * | 2018-08-30 | 2019-01-04 | 金维他(福建)食品有限公司 | β-glucan extracting method in a kind of oat and oatmeal processing byproduct |
| CN114651931A (en) * | 2022-04-01 | 2022-06-24 | 邹阳 | Desugared and defatted wheat bran, oat bran and desugared and defatted method of wheat bran and oat bran |
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| CN103497258A (en) * | 2013-10-11 | 2014-01-08 | 黑龙江八一农垦大学 | Method for extracting and purifying active polysaccharides of wheat bran |
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| CN106520859A (en) * | 2016-10-28 | 2017-03-22 | 淮阴师范学院 | High-yield preparation method for active polysaccharides of wheat bran |
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| CN105384837A (en) * | 2015-12-16 | 2016-03-09 | 黑龙江众生生物工程有限公司 | Microwave-supersonic wave-combined auricularia auriculajudae polysaccharide extraction method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108752491A (en) * | 2018-05-21 | 2018-11-06 | 黑龙江八农垦大学 | Ultrasound-microwave radiation technology water extraction extracts folium isatidis active polysaccharide technique |
| CN108752491B (en) * | 2018-05-21 | 2020-06-09 | 黑龙江八一农垦大学 | Process for extracting folium isatidis active polysaccharide by ultrasonic-microwave-assisted water extraction method |
| CN109134697A (en) * | 2018-08-30 | 2019-01-04 | 金维他(福建)食品有限公司 | β-glucan extracting method in a kind of oat and oatmeal processing byproduct |
| CN114651931A (en) * | 2022-04-01 | 2022-06-24 | 邹阳 | Desugared and defatted wheat bran, oat bran and desugared and defatted method of wheat bran and oat bran |
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