CN106892999A - A kind of brown alga okamura ishige algae extraction method of polysaccharides - Google Patents
A kind of brown alga okamura ishige algae extraction method of polysaccharides Download PDFInfo
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
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Abstract
The present invention relates to a kind of brown alga okamura ishige algae extraction method of polysaccharides, the method is comprised the following steps：High-pressure sterilizing pot is extracted and is combined with ultrasonic wave water extraction by the steps such as collection drying, crushing, pretreatment, mixing extraction, centrifugation, concentrating and precipitating, the present invention, greatly increases the recovery rate of okamura ishige algae polysaccharide；Pre-treatment step of the invention makes the polyoses content that extraction is obtained higher, and the penetration power extracted using water as solvent during polysaccharide to alginic cell is strong, and extraction efficiency is high, and water uses safety, economy in production；The polysaccharide that the present invention is prepared is yellow-white, odorless, with certain gel characteristic.
Description
Technical field
The present invention relates to Polyose extraction field, and in particular to a kind of brown alga okamura ishige algae extraction method of polysaccharides.
Background technology
Algal polysaccharides are a kind of natural active matters, and it has the effect of regulation body's immunity, such as can be by promoting
Enter lymphopoiesis to come with the approach such as differentiation, the phagocytic function of stimulating expression of macrophage, the generation for promoting cell factor and antibody
Realize the regulation to body immune system function.
At present, global most popular marine alga abstraction function food ingredient is exactly to be extracted from the brown algas such as sea-tangle, bulk kelp
The natural seaweed polyose such as alginate.But little, the okamura ishige algae (Ishige of polysaccharide researches is extracted from okamura ishige algae
Okamurai Yendo) it is algae Phaeophyta Ishigeaceae okamura ishige algae platymiscium, it is distributed widely in China East Sea, South Sea hermatolith stone
On, it is among the people for throat inflammation, lymphnoditis, treatment strumous, with clearing heat and detoxicating, the effect of softening and resolving hard mass.It is domestic right
The research of okamura ishige algae is less, and 39 compounds are isolated in the okamura ishige algae that soup seapeak etc. is produced from Zhejiang；The report such as Yang Shangjun is separated
Go out fucosterol, mannitol, hexadecanoic acid；Agglutinin is extracted in the okamura ishige algae that Zheng Yi etc. is produced from Fujian to hay bacillus, beer
Yeast cells has preferable agglutination；Gong Lifen etc. extracts water-soluble polysaccharide with ultrasonic synergistic hot water, has investigated liquid-solid ratio, has surpassed
The influence of intensity of acoustic wave, extraction time, Extracting temperature and pH to okamura ishige algae polysaccharide extract rate, its highest recovery rate is 1.03%,
It is China coast sociales in view of brown alga okamura ishige algae, utilizes natural resources in order to abundant, it is necessary to which phase is carried out to okamura ishige algae
The research of active component is closed, for prior art okamura ishige algae extraction method of polysaccharides, recovery rate is relatively low, it is necessary to increase correlative study,
To improve the recovery rate of polysaccharide.
The content of the invention
In order to solve the problems, such as prior art, the invention provides a kind of brown alga okamura ishige algae extraction method of polysaccharides, should
Method is simple to operate, and using safety, extraction efficiency is high, can be used to mass produce.
A kind of brown alga okamura ishige algae extraction method of polysaccharides, comprises the following steps：
1) collection drying：The flowing water cleaning of collection okamura ishige algae, is cleaned with ultrasonic cleaner low frequency, is put into constant incubator
Drying；
2) crush：Okamura ishige algae sample is crushed with pulverizer, okamura ishige algae dry powder is obtained with after screen filtration；
3) pre-process：Take step 2) in okamura ishige algae dry powder in container, add 95% ethanol, immersion, use double-deck yarn
Cloth is filtered, and is repeated 3 times, and the okamura ishige algae after filtering is air-dried and obtains final product pretreatment okamura ishige algae dry powder；
4) mixing is extracted：Distilled water is added in the okamura ishige algae dry powder of pretreatment, in high-pressure sterilizing pot under the conditions of 115 DEG C
0.5h is extracted, is then placed in that frequency is transferred into 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min in supersonic generator；
5) it is centrifuged：Centrifugation, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and obtains concentrate, is added in concentrate
95% ethanol, stands, and centrifugation is eventually adding 75% ethanol cyclic washing, the precipitation drying for obtaining to okamura ishige algae polysaccharide.
Preferably, the step 1) ultrasonic wave low frequency be 25KHZ；The time of ultrasonic cleaner cleaning for 8~
12min, more preferably 10min.
Preferably, the step 1) in drying procedure first with 120 DEG C of drying 15min, to be then adjusted to 80 DEG C, dry
8 hours to constant weight.
Preferably, the step 2) in screen cloth mesh number be 80~100 mesh.
Preferably, the step 3) okamura ishige algae dry powder and 95% ethanol weight ratio be 1:5, the soak time is
11~13h.
Preferably, the step 4) in the weight ratio of pretreatment okamura ishige algae dry powder and distilled water be 1:5~1:6.
Preferably, the step 5) and step 6) in centrifugal rotational speed be 4000r/min, centrifugation time is 10min.
Preferably, the step 6) in 95% ethanol volume be 3~5 times of concentrate.
Preferably, the step 6) in time of repose be 8~10 hours.
As a kind of perferred technical scheme, a kind of brown alga okamura ishige algae extraction method of polysaccharides, comprises the following steps：
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 10min is cleaned with ultrasonic cleaner low frequency, is put into constant temperature
120 DEG C, 15min in incubator；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, okamura ishige algae dry powder is obtained after crossing 80 mesh sieve net filtrations.
3) pre-process：Take step 2) in okamura ishige algae dry powder in container, add 95% ethanol, okamura ishige algae dry powder with
95% ethanol weight ratio is 1:5,12h is soaked, filtered with double gauze, it is repeated 3 times, the okamura ishige algae after filtering is air-dried and is obtained final product
Pretreatment okamura ishige algae dry powder；
4) mixing is extracted：The weight ratio of distilled water, pretreatment algae powder and distilled water is added in okamura ishige algae dry powder is pre-processed
It is 1:5,0.5h is extracted under the conditions of 115 DEG C in high-pressure sterilizing pot, it is then placed in being transferred to frequency in supersonic generator
130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and obtains concentrate, is added in concentrate
3 times of 95% ethanol of volume, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, is eventually adding
75% ethanol cyclic washing, the precipitation drying for obtaining to okamura ishige algae polysaccharide.
As a kind of perferred technical scheme, a kind of brown alga okamura ishige algae extraction method of polysaccharides, comprises the following steps：
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 12min is cleaned with ultrasonic cleaner low frequency, is put into constant temperature
120 DEG C, 15min in incubator；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, obtaining okamura ishige algae after 100 mesh sieve net filtrations excessively does
Powder.
3) pre-process：Take step 2) in okamura ishige algae dry powder in container, add 95% ethanol, okamura ishige algae dry powder with
95% ethanol weight ratio is 1:5,13h is soaked, filtered with double gauze, it is repeated 3 times, the okamura ishige algae after filtering is air-dried and is obtained final product
Pretreatment okamura ishige algae dry powder；
4) mixing is extracted：Distilled water, pretreatment okamura ishige algae dry powder and distilled water are added in okamura ishige algae dry powder is pre-processed
Weight ratio is 1:5,0.5h is extracted under the conditions of 115 DEG C in the high-pressure sterilizing pot, it is then placed in adjusting frequency in supersonic generator
To 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and obtains concentrate, is added in concentrate
4 times of 95% ethanol of volume, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, is eventually adding
75% ethanol cyclic washing, the precipitation drying for obtaining to okamura ishige algae polysaccharide.
As a kind of perferred technical scheme, a kind of brown alga okamura ishige algae extraction method of polysaccharides, comprises the following steps：
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 8min is cleaned with ultrasonic cleaner low frequency, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, obtaining okamura ishige algae after 100 mesh sieve net filtrations excessively does
Powder.
3) pre-process：Take step 2) in okamura ishige algae dry powder in container, add 95% ethanol, okamura ishige algae dry powder with
95% ethanol weight ratio is 1:5,11h is soaked, filtered with double gauze, it is repeated 3 times, the okamura ishige algae after filtering is air-dried and is obtained final product
Pretreatment okamura ishige algae dry powder；
4) mixing is extracted：Distilled water, pretreatment okamura ishige algae dry powder and distilled water are added in okamura ishige algae dry powder is pre-processed
Weight ratio is 1:5,0.5h is extracted under the conditions of 115 DEG C in high-pressure sterilizing pot, it is then placed in adjusting frequency in supersonic generator
To 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and obtains concentrate, is added in concentrate
5 times of 95% ethanol of volume, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, is eventually adding
75% ethanol cyclic washing, the precipitation drying for obtaining to okamura ishige algae polysaccharide.
High-pressure sterilizing pot can not be diffused into outside based on the steam that water is formed when boiling, and be gathered in the appearance of sealing
In device, in the case of closed, with boiling for water, steam pressure is raised, and temperature also accordingly increases.Ultrasonic wave is a kind of high frequency
The mechanical wave of rate, can form " cavitation " that powerful shock wave or high-speed jet produce and alga cells film is destroyed,
Have the decomposition using active ingredient, can effectively reduce, eliminate with the block layer between water phase, increase mass-transfer efficiency, help
In the diffusion of solute.The simple method for extracting okamura ishige algae polysaccharide using ultrasonic wave water extraction has the simple to operate, time short at present
The advantages of, but recovery rate is not high；Extracted using under the conditions of 115 DEG C of high-pressure sterilizing pot, the percent hydrolysis of frond polysaccharide can be increased, this
High-pressure sterilizing pot is extracted in invention and ultrasonic wave water extraction is combined, and greatly increases the recovery rate of okamura ishige algae polysaccharide；Simultaneously
Pre-treatment step of the invention makes the polyoses content that extraction is obtained higher；The present invention is extracted during polysaccharide to sea using water as solvent
The penetration power of frustule is strong, and extraction efficiency is high, and using safety, economy in production, the polysaccharide that the present invention is prepared is Huang to water
White, odorless, with certain gel characteristic.
Specific embodiment
Using phend-sulphuric acid：With glucose as a standard sample, takes 0.10g polyoses extracts, plus distilled water is dissolved into
After 10.0mL, 1.00mL testing samples are taken out, add the 5.00mL concentrated sulfuric acids, the phenol for adding 1.00mL volume fractions 5% is mixed
Close uniform, develop the color at room temperature, determine its absorbance A at 490nm in 20min, calculate the content of polysaccharide in sample solution.
Okamura ishige algae is locality：Zhoushan saddle archipelago.
Embodiment 1
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 10min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, okamura ishige algae dry powder is obtained after crossing 80 mesh sieve net filtrations.
3) pre-process：Take step 2) in okamura ishige algae dry powder 1KG in container, add 5KG 95% ethanol, immersion
12h, is filtered with double gauze, is repeated 3 times, and the okamura ishige algae after filtering is air-dried and obtains final product pretreatment okamura ishige algae dry powder；
4) mixing is extracted：Distilled water 5L is added in okamura ishige algae dry powder is pre-processed, in the high-pressure sterilizing pot under the conditions of 115 DEG C
0.5h is extracted, is then placed in that frequency is transferred into 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min in supersonic generator；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 600ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains white-yellowish solid 73.8g, determines polyoses content
76.5%.
Embodiment 2
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 12min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, obtaining okamura ishige algae after 100 mesh sieve net filtrations excessively does
Powder.
3) pre-process：Take step 2) in okamura ishige algae dry powder 1KG in container, add 5KG 95% ethanol, immersion
13h, is filtered with double gauze, is repeated 3 times, and the okamura ishige algae after filtering is air-dried and obtains final product pretreatment okamura ishige algae dry powder；
4) mixing is extracted：Distilled water 5L is added in okamura ishige algae dry powder is pre-processed, in the high-pressure sterilizing pot under the conditions of 115 DEG C
0.5h is extracted, is then placed in that frequency is transferred into 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min in supersonic generator；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 650ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains white-yellowish solid 86.2, determines polyoses content 70.4%
g。
Embodiment 3
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 8min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, obtaining okamura ishige algae after 100 mesh sieve net filtrations excessively does
Powder.
3) pre-process：Take step 2) in okamura ishige algae dry powder 1KG in container, add 5KG 95% ethanol, immersion
11h, is filtered with double gauze, is repeated 3 times, and the okamura ishige algae after filtering is air-dried and obtains final product pretreatment okamura ishige algae dry powder；
4) mixing is extracted：Distilled water 5L is added in okamura ishige algae dry powder is pre-processed, in the high-pressure sterilizing pot under the conditions of 115 DEG C
Extract 0.5h, after be put into supersonic generator frequency be transferred to 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 680ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains white-yellowish solid 83.3g, determines polyoses content
72.2%.Comparative example 1
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 10min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, okamura ishige algae dry powder is obtained after crossing 80 mesh sieve net filtrations.
3) pre-process：Take step 2) in okamura ishige algae dry powder 1KG in container, add 5KG 95% ethanol, immersion
12h, is filtered with double gauze, is repeated 3 times, and the frond after filtering is air-dried and obtains final product pretreatment okamura ishige algae dry powder；
4) ultrasonic extraction：Distilled water 5L is added in okamura ishige algae dry powder is pre-processed, is put into frequency in supersonic generator
It is transferred to 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 300ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains brown solid 23.8g, determines polyoses content 50.2%.
Comparative example 2
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 12min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, obtaining okamura ishige algae after 100 mesh sieve net filtrations excessively does
Powder.
3) pre-process：Take step 2) in okamura ishige algae dry powder 1KG in container, add 5KG 95% ethanol, immersion
13h, is filtered with double gauze, is repeated 3 times, and the frond after filtering is air-dried and obtains final product pretreatment okamura ishige algae dry powder；
4) ultrasonic extraction：Distilled water 5L is added in okamura ishige algae dry powder is pre-processed, is put into frequency in supersonic generator
It is transferred to 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 350ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains brown solid 24.9g, determines polyoses content 56.7%.
Comparative example 3
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 8min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, obtaining okamura ishige algae after 100 mesh sieve net filtrations excessively does
Powder.
3) pre-process：Take step 2) in okamura ishige algae dry powder 1KG in container, add 5KG 95% ethanol, immersion
11h, is filtered with double gauze, is repeated 3 times, and the frond after filtering is air-dried and obtains final product pretreatment okamura ishige algae dry powder；
4) ultrasonic extraction：Distilled water 5L is added in okamura ishige algae dry powder is pre-processed, after be put into supersonic generator will frequency
Rate is transferred to 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min；
5) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 350ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains brown solid 23.1g, and determining polyoses content is
50.1%.Comparative example 4
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 10min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, okamura ishige algae dry powder is obtained after crossing 80 mesh sieve net filtrations.
3) mixing is extracted：Take step 2) in okamura ishige algae dry powder 1KG in container, add distilled water 5L, in autoclaving
Extract 0.5h under the conditions of 115 DEG C in pot, after be put into supersonic generator frequency be transferred to 130KHZ, 75 DEG C of temperature, ultrasound extraction
Take 30min；
4) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
5) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 650ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains brown solid 64.8g, determines polyoses content 48.5%.
Comparative example 5
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 12min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, obtaining okamura ishige algae after 100 mesh sieve net filtrations excessively does
Powder.
3) mixing is extracted：Take step 2) in okamura ishige algae dry powder 1KG in container, add distilled water 5L, in autoclaving
Extract 0.5h under the conditions of 115 DEG C in pot, after be put into supersonic generator frequency be transferred to 130KHZ, 75 DEG C of temperature, ultrasound extraction
Take 30min；
4) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
5) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 650ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains brown solid 66.2, determines polyoses content 47.9%g.
Comparative example 6
1) collection drying：The flowing water cleaning of collection okamura ishige algae, 8min is cleaned with ultrasonic cleaner 25KHZ, is put into constant temperature training
Support 120 DEG C, 15min in case；80 DEG C are adjusted to, are dried 8 hours to constant weight.
2) crush：Okamura ishige algae sample is crushed with pulverizer, obtaining okamura ishige algae after 100 mesh sieve net filtrations excessively does
Powder.
3) mixing is extracted：Take step 2) in okamura ishige algae dry powder 1KG in container, add distilled water 5L, in autoclaving
Extract 0.5h under the conditions of 115 DEG C in pot, after be put into supersonic generator frequency be transferred to 130KHZ, 75 DEG C of temperature, ultrasound extraction
Take 30min；
4) it is centrifuged：Centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, collects supernatant；
5) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate 660ml is obtained, to concentrate
3 times of 95% ethanol of volume of middle addition, stand 8 hours, centrifugation, and centrifugal rotational speed is 4000r/min, and centrifugation time is 10min, most
75% ethanol cyclic washing is added afterwards, and the precipitation drying for obtaining obtains brown solid 66.3g, determines polyoses content 47.8%.
This place embodiment is in place of the claimed non-limit of technical scope midrange and in embodiment technology
The new technical scheme formed to the equal replacement of single or multiple technical characteristics in scheme, equally all in application claims
In the range of protection；Simultaneously the present invention program it is all enumerate or unrequited embodiment in, in the same embodiment each
Parameter is merely representative of an example (i.e. a kind of feasible scheme) of its technical scheme, and between parameters and in the absence of strict
Cooperation and qualified relation, wherein each parameter can be replaced mutually when stating and asking without prejudice to axiom and the present invention, special declaration
Except.
Technological means disclosed in the present invention program is not limited only to the technological means disclosed in above-mentioned technological means, also includes
Constituted technical scheme is combined by above technical characteristic.The above is specific embodiment of the invention, should be referred to
Go out, for those skilled in the art, under the premise without departing from the principles of the invention, can also make some
Improvements and modifications, these improvements and modifications are also considered as protection scope of the present invention.
Claims (9)
1. a kind of brown alga okamura ishige algae extraction method of polysaccharides, it is characterised in that comprise the following steps：
1) collection drying：The flowing water cleaning of collection okamura ishige algae, is cleaned with ultrasonic cleaner low frequency, is put into constant incubator baking
It is dry；
2) crush：Okamura ishige algae sample is crushed with pulverizer, okamura ishige algae dry powder is obtained with after screen filtration；
3) pre-process：Take step 2) in okamura ishige algae dry powder in container, add 95% ethanol, immersion, use double gauze mistake
Filter, is repeated 3 times, and the okamura ishige algae after filtering is air-dried and obtains final product pretreatment okamura ishige algae dry powder；
4) mixing is extracted：Distilled water is added in okamura ishige algae dry powder is pre-processed, is extracted under the conditions of 115 DEG C in the high-pressure sterilizing pot
0.5h, is then placed in that frequency is transferred into 130KHZ, 75 DEG C of temperature, ultrasonic extraction 30min in supersonic generator；
5) it is centrifuged：Centrifugation, collects supernatant；
6) concentrating and precipitating：Vacuum rotary evaporator has been concentrated into solid and has started to separate out, and concentrate is obtained, to adding 95% in concentrate
Ethanol, stands, and centrifugation is eventually adding 75% ethanol cyclic washing, the precipitation drying for obtaining to okamura ishige algae polysaccharide.
2. a kind of brown alga okamura ishige algae extraction method of polysaccharides according to claim 1, it is characterised in that the step 1) in ultrasound
Ripple low frequency is 25KHZ；The time of ultrasonic cleaner cleaning is 8~12min, more preferably 10min.
3. a kind of brown alga okamura ishige algae extraction method of polysaccharides according to claim 1, it is characterised in that the step 1) in baking
Dry program is 120 DEG C of drying 15min in first using, and is then adjusted to 80 DEG C, is dried 8 hours to constant weight.
4. a kind of brown alga okamura ishige algae extraction method of polysaccharides according to claim 1, it is characterised in that the step 2) in screen cloth
Mesh number be 80~100 mesh.
5. a kind of brown alga okamura ishige algae extraction method of polysaccharides according to claim 1, it is characterised in that the step 3) iron nail
Dried vegetable powder and 95% ethanol weight ratio are 1:5, the soak time is 11~13h.
6. a kind of brown alga okamura ishige algae extraction method of polysaccharides according to claim 1, it is characterised in that the step 4) in it is pre-
Treatment okamura ishige algae dry powder is 1 with the weight ratio of distilled water:5~1:6.
7. a kind of brown alga okamura ishige algae extraction method of polysaccharides according to claim 1, it is characterised in that the step 5) and step
6) centrifugal rotational speed in is 4000r/min, and centrifugation time is 10min.
8. a kind of brown alga okamura ishige algae extraction method of polysaccharides according to claim 1, it is characterised in that the step 6) in 95%
The volume of ethanol be 3~5 times of concentrate.
9. a kind of brown alga okamura ishige algae extraction method of polysaccharides according to claim 1, it is characterised in that the step 6) in stand
Time is 8~10 hours.
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