CN106832022A - A kind of method for extracting fucoidan - Google Patents
A kind of method for extracting fucoidan Download PDFInfo
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- CN106832022A CN106832022A CN201510889701.XA CN201510889701A CN106832022A CN 106832022 A CN106832022 A CN 106832022A CN 201510889701 A CN201510889701 A CN 201510889701A CN 106832022 A CN106832022 A CN 106832022A
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- fucoidan
- shredded kelp
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Abstract
The invention discloses a kind of method for extracting fucoidan, comprise the following steps：Sea-tangle is cut into uniform Shredded kelp, Shredded kelp is entered into water-filling after being processed with ethanol and boils immersion, extracted through acid again and microwave counter current is extracted, obtain Fucoidan sulfuric ester crude extract, calcium chloride is added, filtrate is obtained after multistage filtering, filtrate is through being concentrated by ultrafiltration to obtain concentrate, drying and crushing after concentrate alcohol precipitation, obtains final product fucoidin sulfuric ester.The inventive method is simple, and, using less, step is few, low cost, and sea-tangle can recycle, and be suitable to industrialized production for organic solvent；Gained marine alga glycan sulfate content is 2.0%-2.8%, and wherein fucose content is 75.2%-80.8%, and sulfate radical content is 55.6%-75.4%.
Description
Technical field
The present invention relates to a kind of extracting method of the active material from sea-tangle, and in particular to one kind is extracted from sea-tangle
The method of fucoidan.
Background technology
Fucoidan, is a kind of water miscible heteromeric polysaccharide also known as fucoidin, its it is main effectively into
Part is α-L-fucose -4- sulfuric esters.Fucoidan is a kind of excellent healthy food, pharmaceutical raw material,
Because naturally containing sulfate radical, so that the characteristic with anionic polymer compound, research shows, fucosan
Sulfuric ester has no toxic side effect, with multiple biological activities including anticoagulation, reducing blood lipid, anti-chronic kidney hypofunction, anti-
Tumour, antiviral, anti-inflammatory, anti-oxidative damage, promotion organization regeneration, suppression gastric ulcer, enhancing immunity of organism
Function and surgical intervention potentiality etc..
Fucoidan is a kind of high-valued product, is the outlet of the high-valued development of sea-tangle marine alga, current rock
The extracting method of algae glycan sulfuric ester has hydro-thermal extraction method, acid extraction method, ultrasonic extraction etc., hydro-thermal formulation
It is, using fucoidan property soluble in water, sour water occurs this method avoid fucoidan
The possibility of solution, can keep the macromolecular structure of fucoidin, have the disadvantage that extracting liquid volume is excessive, in extract
There is more alginic acid, increased the difficulty for isolating and purifying；Acid extraction method is to utilize alginic acid at a low ph
The property for dissolving is difficult to, fucoidan is extracted with watery hydrochloric acid, the method has extracting cycle
The shortcomings of length, big energy-consuming；Ultrasonic extraction method polysaccharide is a kind of Polyose extraction new method for getting a good chance of, can be with
Improve extraction rate and shorten extraction time, it is prolonged to make because there is ultrasonic wave stronger mechanical shearing to act on
With the Fragmentation of polysaccharides that can make macromolecular, the proposition of polysaccharide is influenceed so as to the loss increase during post processing
Rate.
Patent ZL98110253.0 discloses a kind of polyfucose sulfate with varible molecular weight (fucoidin)
Preparation method, patent ZL 200410023694 discloses a kind of system of fucosan sulfate of high sulfuric content
Preparation Method, but these methods are that fucoidan is extracted with hot water and acid adding, and recovery rate is not high, and
And pollution environment.Patent 200510047582.X extracts fucoidan with enzymatic isolation method, however it is necessary that with
To expensive cellulase and pectase, production cost is caused to improve.
Traditional heat is extracted to be carried out from outside to inside with modes such as heat transfer, heat radiations, and Microwave Extraction is by idol
Extremely son rotation and ionic conduction two ways are inside and outside heated simultaneously, and Microwave Extraction has the characteristics that：Selectivity height,
Operating time is short, solvent-oil ratio is few, low energy consumption, pollution-free, effective component yield is high and does not produce noise
Deng；Continuous Countercurrent Extraction refers to that during extraction, material and solvent are continuously moved simultaneously, but the direction of motion
Conversely, by mechanical transfer, continuous and quantitative charging is fully contacted material and solvent, and equipment internal solvent is continuous
Update, finally continuously slag tap, while constantly updating solvent in device interior, solvent is constantly obtained in flow process
Material effective ingredient, concentration improve constantly, continuous feed liquor and continuously go out liquid during, deposited in solvent
In continuous concentration gradient, so that extract solution can be obtained than leaching velocity faster, it is also possible to obtain compare
Extract concentration high.
The content of the invention
In order to solve problem of the prior art, the invention provides a kind of simple to operate, low cost, high income,
Purity is high
Fucoidan method for extraction and purification.
The technical scheme that the present invention is used to achieve the above object is comprised the following steps：
1) dry sea-tangle is chosen, sea-tangle is cut into the Shredded kelp of 0.2-0.4mm after cleaning up；
The step 1) in the selection frond plumpness suitable limnetic dry kelp of maturity, remove root, yellowish-white side and
The position of scabbing of rotting and hole, is cut into uniform Shredded kelp, helps to extract fucoidan,
It is beneficial to recycle.
2) it is 95% ethanol solution the Shredded kelp of above-mentioned 1 weight portion to be added into the mass concentration of 10-20 weight portions
In, stirring, 40-60 DEG C of extraction 0.5-1h separates to obtain Shredded kelp A and ethanol extract；
The step 2) in ethanol can remove pigment in sea-tangle, ethanol extract can reclaim mannitol and
Iodine.
3) after the Shredded kelp A of 1 weight portion is rinsed 2-4 minutes with clear water, the water-soluble of 3-6 weight portions is added
Liquid extracted after soaking water is boiled, and aqueous extract and Shredded kelp B is collected by centrifugation；
The step 3) in add the aqueous solution first to soak 2-10h, then heated at constant temperature is to 20-90 DEG C, heat time
It is 0.5-2h, is incubated 1-2h, soaks 3-24h after natural cooling again, it is soluble in water using fucoidan
Property, there is acid-hydrolyzed possibility this method avoid fucoidan, fucosan sulphur can be kept
The macromolecular structure of acid esters, by the multiple immersion and heating of the aqueous solution, can make more fucosan sulphur
Acid esters is soluble in water.
4) solvent is added in Shredded kelp B, microwave counter current extraction is carried out under 70-90 DEG C of constant temperature, microwave power is
400-800W, extraction time is 50-70min, and after the completion of extraction, extract solution is through being centrifuged to obtain supernatant and sea-tangle
Silk C, centrifugal speed is 500-1500r/min, and the time is 20-40min, and aqueous extract and supernatant merging are
Fucoidan crude extract；
The step 4) in, raw material is 1 with the mass ratio of solvent:4-10.
Preferably, the raw material and the mass ratio of solvent are 1:6.
The step 4) in solvent can be deionized water, low concentration acid or low concentration acid and organic acid
Mixing, step 4) described in low concentration acid for 0.08-0.15mol/L hydrochloric acid, 0.06-0.12mol/L nitre
Acid or 0.10-0.18mol/L sulfuric acid in one kind, the organic acid be acetic acid or carboxylic acid, the organic acid it is dense
It is 0.05-0.10mol/L to spend.
Preferably, solvent is the mixing of the hydrochloric acid and 0.05-0.10mol/L acetic acid of 0.08-0.15mol/L.
5) excess chlorination calcium is added in fucoidan crude extract, is occurred to without precipitation, stand 12-26
Hour, then carry out multistage filtering and obtain filtrate；
The step 5) in multistage filtering include heat filtering, centrifugation and ultrafiltration, heat filtering temperature be 70-85 DEG C,
Centrifugal rotational speed is first 1000-2000r/min, and centrifugal speed is adjusted to 2000-4000r/min after continuing 10-30min,
Continue 10-30min, ultrafiltration is the milipore filter for 3000-5000kDa by molecular cut off.
6) filtrate is concentrated using milipore filter ultrafiltration, is obtained concentrate.
The step 6) in milipore filter for molecular cut off 1000-2000 hollow cellulose milipore filter.
7) concentrate of 1 weight portion is added to the mass concentration of 2-5 weight portions in 95% ethanol solution,
Being placed at 5 DEG C makes precipitation completely, and precipitation is dried after centrifugation, obtains final product fucoidin sulfuric ester sterling；
8) reclaimed after the cleaned dryings of Shredded kelp C.
Compared with prior art, beneficial effects of the present invention are：
1) the inventive method is simple, and, using less, step is few, low cost, and sea-tangle can recycle for organic solvent,
It is suitable to industrialized production；
2) extracted using water, acid is extracted and microwave counter current extracting method improves product yield.
3) present invention is concentrated by ultrafiltration method in the purge process of fucoidan using multistage filtering, makes
Product purity is greatly improved, and fucoidan content is 2.0%-2.8%, and wherein fucose content is
75.2%-80.8%, sulfate radical content is 55.6%-75.4%.
Specific embodiment
Embodiment 1
The Shredded kelp of 1 weight portion is added into the mass concentration of 10 weight portions in 95% ethanol solution, to stir,
40 DEG C of extraction 0.5h, separate to obtain Shredded kelp A and ethanol extract；By the Shredded kelp A clear water of 1 weight portion
After rinsing 3 minutes, the aqueous solution soaking 2h of 3 weight portions, then heated at constant temperature are added to 20 DEG C, heat time
It is 0.5h, is incubated 1h, soak 3h after natural cooling again, aqueous extract and Shredded kelp B is collected by centrifugation；1
The mixing of the hydrochloric acid and 0.05mol/L acetic acid of the 0.08mol/L of 6 weight portions is added in the Shredded kelp B of weight portion
Solvent, carries out microwave counter current extraction under 70 DEG C of constant temperature, microwave power is 400W, and extraction time is 50min,
After the completion of extraction, extract solution is 500r/min, time through supernatant and Shredded kelp C, centrifugal speed is centrifuged to obtain
It is 20min, aqueous extract and supernatant merging are fucoidan crude extract, and Shredded kelp C is cleaned
Dried recovered is utilized afterwards；Excess chlorination calcium is added in fucoidan crude extract, is occurred to without precipitation,
12 hours are stood, heat filtering, centrifugation and ultrafiltration are being carried out successively, heat filtering temperature is 70 DEG C, centrifugal rotational speed
First be 1000r/min, continue 10min after centrifugal speed be adjusted to 2000r/min, continue 10min, ultrafiltration be through
The milipore filter that molecular cut off is 3000 is crossed, filtrate is obtained；The hollow fibre that filtrate is passed through into molecular cut off 1000
The plain milipore filter of dimension is concentrated, and obtains concentrate；The quality that the concentrate of 1 weight portion is added into 2 weight portions is dense
In spending for 95% ethanol solution, being placed at 5 DEG C makes precipitation completely, and precipitation is dried after centrifugation, obtains final product rock algae
Polysaccharide sulfate sterling, fucoidan content is 2.0%, and wherein fucose content is 75.5%, sulphur
Acid group content is 55.6%.
Embodiment 2
The Shredded kelp of 1 weight portion is added into the mass concentration of 20 weight portions in 95% ethanol solution, to stir,
60 DEG C of extraction 1h, separate to obtain Shredded kelp A and ethanol extract；The Shredded kelp A of 1 weight portion is rushed with clear water
After washing 3 minutes, the aqueous solution soaking 10h of 6 weight portions, then heated at constant temperature are added to 90 DEG C, the heat time is
02h, is incubated 2h, soaks 24h after natural cooling again, and aqueous extract and Shredded kelp B is collected by centrifugation；In 1 weight
Add the mixing of the hydrochloric acid and 0.10mol/L acetic acid of the 0.15mol/L of 6 weight portions molten in the Shredded kelp B for measuring part
Agent, carries out microwave counter current extraction under 90 DEG C of constant temperature, microwave power is 800W, and extraction time is 70min, is carried
After the completion of taking, through supernatant and Shredded kelp C is centrifuged to obtain, centrifugal speed is 1500r/min to extract solution, and the time is
40min, aqueous extract and supernatant merging are fucoidan crude extract, and Shredded kelp C is once purged
Dried recovered is utilized；Excess chlorination calcium is added in fucoidan crude extract, is occurred to without precipitation,
26 hours are stood, heat filtering, centrifugation and ultrafiltration are being carried out successively, heat filtering temperature is 85 DEG C, centrifugal rotational speed
First be 2000r/min, continue 30min after centrifugal speed be adjusted to 4000r/min, continue 30min, ultrafiltration be through
The milipore filter that molecular cut off is 3000 is crossed, filtrate is obtained；The hollow fibre that filtrate is passed through into molecular cut off 1000
The plain milipore filter of dimension is concentrated, and obtains concentrate；The quality that the concentrate of 1 weight portion is added into 5 weight portions is dense
In spending for 95% ethanol solution, being placed at 5 DEG C makes precipitation completely, and precipitation is dried after centrifugation, obtains final product rock algae
Polysaccharide sulfate sterling, fucoidan yield is 2.5%, and wherein fucose content is 78.2%, sulphur
Acid group content is 58.6%.
Embodiment 3
The Shredded kelp of 1 weight portion is added into the mass concentration of 15 weight portions in 95% ethanol solution, to stir,
50 DEG C of extraction 1h, separate to obtain Shredded kelp A and ethanol extract；The Shredded kelp A of 1 weight portion is rushed with clear water
After washing 3 minutes, the aqueous solution soaking 8h of 6 weight portions, then heated at constant temperature are added to 80 DEG C, the heat time is
1h, is incubated 1.5h, soaks 12h after natural cooling again, and aqueous extract and Shredded kelp B is collected by centrifugation；In 1 weight
Add the mixing of the hydrochloric acid and 0.08mol/L acetic acid of the 0.10mol/L of 6 weight portions molten in the Shredded kelp B for measuring part
Agent, carries out microwave counter current extraction under 80 DEG C of constant temperature, microwave power is 600W, and extraction time is 60min, is carried
After the completion of taking, through supernatant and Shredded kelp C is centrifuged to obtain, centrifugal speed is 1000r/min to extract solution, and the time is
30min, aqueous extract and supernatant merging are fucoidan crude extract, and Shredded kelp C is once purged
Dried recovered is utilized；Excess chlorination calcium is added in fucoidan crude extract, is occurred to without precipitation,
18 hours are stood, heat filtering, centrifugation and ultrafiltration are being carried out successively, heat filtering temperature is 78 DEG C, centrifugal rotational speed
First be 1500r/min, continue 20min after centrifugal speed be adjusted to 3200r/min, continue 20min, ultrafiltration be through
The milipore filter that molecular cut off is 3000 is crossed, filtrate is obtained；The hollow fibre that filtrate is passed through into molecular cut off 1000
The plain milipore filter of dimension is concentrated, and obtains concentrate；The quality that the concentrate of 1 weight portion is added into 3 weight portions is dense
In spending for 95% ethanol solution, being placed at 5 DEG C makes precipitation completely, and precipitation is dried after centrifugation, obtains final product rock algae
Polysaccharide sulfate sterling, fucoidan yield is 2.8%, and wherein fucose content is 80.8%, sulphur
Acid group content is 75.4%.
Embodiment 4
The Shredded kelp of 1 weight portion is added into the mass concentration of 16 weight portions in 95% ethanol solution, to stir,
60 DEG C of extraction 1h, separate to obtain Shredded kelp A and ethanol extract；The Shredded kelp A of 1 weight portion is rushed with clear water
After washing 3 minutes, the aqueous solution soaking 7h of 4 weight portions, then heated at constant temperature are added to 60 DEG C, the heat time is
1.2h, is incubated 2h, soaks 16h after natural cooling again, and aqueous extract and Shredded kelp B is collected by centrifugation；In 1 weight
Add the mixing of the hydrochloric acid and 0.10mol/L acetic acid of the 0.10mol/L of 6 weight portions molten in the Shredded kelp B for measuring part
Agent, carries out microwave counter current extraction under 78 DEG C of constant temperature, microwave power is 700W, and extraction time is 55min, is carried
After the completion of taking, through supernatant and Shredded kelp C is centrifuged to obtain, centrifugal speed is 800r/min to extract solution, and the time is
20min, aqueous extract and supernatant merging are fucoidan crude extract, and Shredded kelp C is once purged
Dried recovered is utilized；Excess chlorination calcium is added in fucoidan crude extract, is occurred to without precipitation,
14 hours are stood, heat filtering, centrifugation and ultrafiltration are being carried out successively, heat filtering temperature is 75 DEG C, centrifugal rotational speed
First be 1200r/min, continue 15min after centrifugal speed be adjusted to 3800r/min, continue 15min, ultrafiltration be through
The milipore filter that molecular cut off is 3000 is crossed, filtrate is obtained；The hollow fibre that filtrate is passed through into molecular cut off 1000
The plain milipore filter of dimension is concentrated, and obtains concentrate；The quality that the concentrate of 1 weight portion is added into 4 weight portions is dense
In spending for 95% ethanol solution, being placed at 5 DEG C makes precipitation completely, and precipitation is dried after centrifugation, obtains final product rock algae
Polysaccharide sulfate sterling, fucoidan yield is 2.5%, and wherein fucose content is 80.6%, sulphur
Acid group content is 65.4%.
The inventive method is simple, and, using less, step is few, low cost, is suitable to industrialized production for organic solvent；Adopt
Extracted with water, acid is extracted and microwave counter current extracting method improves product yield, in the pure of fucoidan
Method is concentrated by ultrafiltration using multistage filtering during change, product purity is greatly improved, fucoidan contains
It is 2.0%-2.8% to measure, and wherein fucose content is 75.2%-80.8%, and sulfate radical content is 55.6%-75.4%.
Clear, complete description, described implementation will be carried out to the technical scheme in the embodiment of the present invention above
Example is a part of embodiment of the invention, rather than whole embodiments, based on the embodiment in the present invention, this
The every other embodiment that field those of ordinary skill is obtained on the premise of creative work is not made, all
Belong to the scope of protection of the invention.
Claims (10)
1. it is a kind of extract fucoidan method, it is characterised in that：Comprise the following steps：
1) dry sea-tangle is chosen, sea-tangle is cut into the Shredded kelp of 0.2-0.4mm after cleaning up；
2) it is 95% ethanol solution the Shredded kelp of above-mentioned 1 weight portion to be added into the mass concentration of 10-20 weight portions
In, stirring, 40-60 DEG C of extraction 0.5-1h separates to obtain Shredded kelp A and ethanol extract；
3) after the Shredded kelp A of 1 weight portion is rinsed 2-4 minutes with clear water, the water-soluble of 3-6 weight portions is added
Liquid extracted after soaking water is boiled, and aqueous extract and Shredded kelp B is collected by centrifugation；
4) solvent is added in Shredded kelp B, microwave counter current extraction is carried out under 70-90 DEG C of constant temperature, microwave power is
400-800W, extraction time is 50-70min, and after the completion of extraction, extract solution is through being centrifuged to obtain supernatant and sea-tangle
Silk C, centrifugal speed is 500-1500r/min, and the time is 20-40min, and aqueous extract and supernatant merging are
Fucoidan crude extract；
5) add excess chlorination calcium to occur to without precipitation in fucoidan crude extract, stand 12-26
Hour, then carry out multistage filtering and obtain filtrate；
6) filtrate is concentrated using milipore filter ultrafiltration, is obtained concentrate；
7) concentrate of 1 weight portion is added to the mass concentration of 2-5 weight portions in 95% ethanol solution,
Being placed at 5 DEG C makes precipitation completely, and precipitation is dried after centrifugation, obtains final product fucoidin sulfuric ester sterling；
8) reclaimed after the cleaned dryings of Shredded kelp C.
2. according to claim 1 it is a kind of extract fucoidan method, it is characterised in that：It is described
Step 3) in add the aqueous solution first soak 2-10h, then heated at constant temperature to 20-90 DEG C, the heat time is 0.5-2h,
Insulation 1-2h, soaks 3-24h again after natural cooling.
3. according to claim 1 it is a kind of extract fucoidan method, it is characterised in that step
4) in, raw material is 1 with the mass ratio of solvent:4-10.
4. according to claim 3 it is a kind of extract fucoidan method, it is characterised in that step
4) solvent in is deionized water, the acid and the mixing of organic acid of the sour or low concentration of low concentration.
5. according to claim 4 it is a kind of extract fucoidan method, it is characterised in that step
4) acid of low concentration described in is the hydrochloric acid of 0.08-0.15mol/L, 0.06-0.12mol/L nitric acid or
One kind in 0.10-0.18mol/L sulfuric acid.
6. according to claim 5 it is a kind of extract fucoidan method, it is characterised in that step
4) organic acid in is acetic acid or carboxylic acid, and the concentration of the organic acid is 0.05-0.10mol/L.
7. according to claim 1 it is a kind of extract fucoidan method, it is characterised in that：Step
5) multistage filtering in includes heat filtering, centrifugation and ultrafiltration.
8. according to claim 1 or 7 it is a kind of extract fucoidan method, it is characterised in that：
Step 5) in heat filtering temperature be 70-85 DEG C, centrifugal rotational speed first be 1000-2000r/min, continue 10-30min
Centrifugal speed is adjusted to 2000-4000r/min afterwards, continues 10-30min.
9. according to claim 7 it is a kind of extract fucoidan method, it is characterised in that：Step
5) ultrafiltration is the milipore filter for 3000-5000 by molecular cut off in.
10. according to claim 1 it is a kind of extract fucoidan method, it is characterised in that：Step
It is rapid 6) in milipore filter for molecular cut off 1000-2000 hollow cellulose milipore filter.
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| WO2020019078A1 (en) * | 2018-07-27 | 2020-01-30 | Arc Medical Devices Inc. | High-molecular-weight fucans for treating fibrous adhesions and other diseases and conditions |
| CN111154005A (en) * | 2019-07-05 | 2020-05-15 | 山东省科学院生物研究所 | Combined extraction method of fucoidin |
| CN112175107A (en) * | 2020-10-16 | 2021-01-05 | 中国海洋大学 | Method for extracting fucosan sulfate and chondroitin sulfate from sea cucumber |
| CN112679627A (en) * | 2021-01-12 | 2021-04-20 | 大连海洋大学 | Method for extracting high-purity fucoidan from brown algae |
| CN113558165A (en) * | 2021-08-24 | 2021-10-29 | 江苏南大耐雀生物技术有限公司 | Preparation process of high-content fucoidin sulfate solid beverage |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020019078A1 (en) * | 2018-07-27 | 2020-01-30 | Arc Medical Devices Inc. | High-molecular-weight fucans for treating fibrous adhesions and other diseases and conditions |
| CN112533989A (en) * | 2018-07-27 | 2021-03-19 | Arc医疗器械股份有限公司 | Highly purified and/or modified fucoidan composition for treating fibrous adhesions |
| US11628183B2 (en) | 2018-07-27 | 2023-04-18 | ARC Medical Ine. | Highly purified fucans for the treatment of fibrous adhesions |
| US11642368B2 (en) | 2018-07-27 | 2023-05-09 | ARC Medical Inc. | Highly purified and/or modified fucan compositions for the treatment of fibrous adhesions |
| US11938145B2 (en) | 2018-07-27 | 2024-03-26 | ARC Medical Inc. | Low endotoxin fucan compositions, systems, and methods |
| CN111154005A (en) * | 2019-07-05 | 2020-05-15 | 山东省科学院生物研究所 | Combined extraction method of fucoidin |
| CN112175107A (en) * | 2020-10-16 | 2021-01-05 | 中国海洋大学 | Method for extracting fucosan sulfate and chondroitin sulfate from sea cucumber |
| CN112175107B (en) * | 2020-10-16 | 2021-08-03 | 中国海洋大学 | Method for extracting fucosan sulfate and chondroitin sulfate from sea cucumber |
| CN112679627A (en) * | 2021-01-12 | 2021-04-20 | 大连海洋大学 | Method for extracting high-purity fucoidan from brown algae |
| CN113558165A (en) * | 2021-08-24 | 2021-10-29 | 江苏南大耐雀生物技术有限公司 | Preparation process of high-content fucoidin sulfate solid beverage |
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Application publication date: 20170613 |