CN106832036A - The preparation method of algal polysaccharides extract - Google Patents
The preparation method of algal polysaccharides extract Download PDFInfo
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Abstract
The present invention proposes a kind of preparation method of algal polysaccharides extract, comprises the following steps：1) dried after marine alga is cleaned, through with mechanical crushing, then with cyclic ultrasonic breaking, obtain marine algae powder；2) by step 1) powder be put into extractor to stir and extracted using pot group type dynamic countercurrent extracting method, obtain extract solution and marine alga waste residue；3) by step 2) marine alga waste residue be put into 60~100 DEG C of hot water, carry out ultrasonic wave extraction, obtain leaching liquor；4) by step 2) obtain extract solution and step 3) leaching liquor insert in stirring reaction container, regulation stirred reactor in Extracting temperature be 20~60 DEG C, pH value be 5~8；5) to step 4) stirring reaction container in add 600~900ppm flocculant, its stirring and flocculation extraction time be 20~30min；6) after the completion of flocculation is extracted, flocculation extract solution is clarified or centrifugal filtration, is obtained algal polysaccharides crude product.The preparation method rate of recovery is high, is easy to industrialized production.
Description
Technical field
The invention belongs to marine active substance extractive technique field, and in particular to a kind of preparation side of algal polysaccharides extract
Method.
Background technology
Ocean area is vast, and marine algae plant origin enriches, and algal polysaccharides are just progressively main next as biological polyoses
One of source.Because marine alga living environment is special, algal polysaccharides have some unique physiologically actives, such as：Laminarin can be reduced
Spleen endoperoxide content, increases the activity of superoxide dismutase (SOD) and catalase (CAT), reaches antitumor
Purpose；Laver amylose, sulfuric acid mannose ester etc. have antithrombotic, improve the effect such as microcirculation；Laminarin, laver amylose etc. are right
Body cell has defencive function.Before increasing result of study has shown that algal polysaccharides have wide development and application
Scape, algal polysaccharide be important composition therein partly, it include fucoidin (Fucoidan), algin (Algin) and brown alga
Starch (Laminaran).Fucoidin is water-soluble polysaccharide, is present in iuntercellular tissue, is mixed to constitute similar heteroglycan
Compound, its main component is L-fucose -4- sulfuric esters, also with a small amount of galactolipin, mannose, xylose, arabinose, sugar
Aldehydic acid etc., fucoidin is generally combined with protein, more difficult (the bright marquis Beijing of marine alga chemistry discipline of grading purification：Scientific publication
Society .1997.p12-17), the derivative of fucoidin gradually shows at the aspect such as anti AIDS virus, treatment kidney failure, hypoglycemic
Show great potential.
The step of extraction and precipitation in algal polysaccharides extraction process are two most criticals.Extraction be by marine alga machinery or
After chemical method is broken, under normal temperature or heating condition, soaked with alkalescence, neutral or acid solution, but when generally there are extraction
Between it is more long, the problems such as leaching recovery is relatively low.Some can be taken to pre-process and purification method reduction leaching liquor before extraction
In other impurity contents, such as pre-processed with Sevage methods, high concentration alcohols and formalin and remove protein, or using acid
Or alkali process frond, but pretreatment often causes polysaccharide loss, reduction polysaccharide recovery.
Obtain containing algal polysaccharides leaching liquor after, the general method using extract solution concentrated under reduced pressure reduces follow-up treatment
Volume, or with dialysis method concentrated extracting solution.Also there is the method (week that extract solution and press filtration are concentrated by evaporation using water-bath under normal pressure
Will just etc., the separation of spirulina maxim polysaccharide, purifying and its oxidation resistant characteristic, Botany Gazette, 1997,39 (1)：77-81；Record
Bright marquis etc., the architectural feature of several glycan of red algae sulfur-bearing half, Oceanologia et Limnologia Sinica, 1996,27 (3) are studied with 13C-NMR spectroscopic methodologies：
330-335)。
From leaching liquor extract separating marine bioactive polysaccharide be nearly all at present to use the precipitation method, such as ethanol precipitation,
Cetylpyridinium chloride (CPC) or cetrimonium bromide (CTAB) precipitation etc..And the extraction of algal polysaccharides is separated
Present is nearly all ethanol precipitation, such as sea grass polysaccharide, spirulina polysaccharide, spirulina mucopolysaccharide, algal polysaccharide sulfate, centipede
Algae carragheen etc. is extracted using ethanol precipitation.Such as, B.Ray etc. is connected using water-boiling method from sea lettuce (Ulva rigida)
Continuous extraction, ethanol precipitation are extracted and separate cell wall polysaccharides (1995)；Zhang Erxian etc. uses second after being boiled through water from sargassum thunbergii and Sargassum horneri
The alcohol precipitation method are extracted and separate algal polysaccharides (1995)；Zhou Zhigang etc. boils with distilled water water-and ethanol precipitation extracts and separated culture
Three kinds (it is very big, it is blunt top and salt pool) the extracellular of spirulina, intracellular polyses (1997)；The general extraction of spirulina polysaccharide is separated
It is after marine alga is extracted through 80 DEG C of hot water, (Xu Hui etc. to be separated with 95% ethanol precipitation：Chinese biochemical drug magazine, 1997,18,
72；Once and equality：Medicine journal, 1995,330,858；A left side continues remote etc.：Pharmaceutical Biotechnology, 1996,3,158).
Ethanol precipitation be although easy to large-scale production, but it there are the following problems：
1st, the relatively low crude product of polyoses content (＜ 50%) typically is can only obtain, because product is containing a large amount of denatured protein etc.
Impurity, increased the difficulty of the purge processes such as follow-up ethanol reprecipitation, ion exchange, chromatography；
2nd, per kilogram Thick many candies need to consume 50-60 kilograms of industrial alcohol, and overall recovery is typically only 10-30%, cause
Product cost is high；
3rd, the investment of ethanol recovery system is big, flow is more complicated, high energy consumption；And whole extraction process when engineering is amplified not
Easily realize serialization, automation.
Although having carried out the research of some other methods at present, but still it is the precipitation method, such as cetylpyridinium chloride (CPC)
Precipitation, cetrimonium bromide (CTAB) precipitation, aluminium hydroxide and lead hydroxide complex compound sediment etc., equally exist with
Ethanol precipitation identical problem.
The recycling that flocculation of electric field is used for bioactivator is a noticeable topic.(the Kumay such as Kumay
H D et.al.Electrical flocculation ofthe unicellular green Algachlorella
Vulgaris.Aquatic Botany.1981,11：Found when 187-195) studying flocculation of electric field monoplast green alga：Electric current passes through
During suspension；Green alga cohesion is floated；3V/cm electric fields, 30 minutes, during pH value=7.0, nearly 90% green alga can be separated from the water；
Zhao Bing et al. (ZL90100012.4,1990) is extracted AMS, alkali protease and carbohydrase using flocculation of electric field method.
But the technique that so far, there is no flocculation to extract algal polysaccharides.
The content of the invention
The rate of recovery existed to solve existing algal polysaccharides to extract is low, and the low problem of activity, and the present invention proposes one
The preparation method of algal polysaccharides extract is planted, the preparation method is easy to industrialized production.
The technical proposal of the invention is realized in this way：
A kind of preparation method of algal polysaccharides extract, comprises the following steps：
1) dried after marine alga is cleaned, through with mechanical crushing, then ensure that alginic cell is broken completely with cyclic ultrasonic breaking
It is bad, obtain marine algae powder；
2) by step 1) powder be put into extractor to stir and extracted using pot group type dynamic countercurrent extracting method, obtain
To extract solution and marine alga waste residue；
3) by step 2) marine alga waste residue be put into 60~100 DEG C of hot water, carry out ultrasonic wave extraction, obtain leaching liquor；
4) by step 2) obtain extract solution and step 3) leaching liquor insert in stirring reaction container, adjust stirring reaction
Extracting temperature is 20~60 DEG C in device, and pH value is 5~8；
5) to step 4) stirring reaction container in add 600~900ppm flocculant, its stirring and flocculation extract when
Between be 20~30min；
6) after the completion of flocculation is extracted, flocculation extract solution is clarified or centrifugal filtration, is obtained algal polysaccharides crude product；
7) the algal polysaccharides crude product that will be obtained is further purified, and obtains algal polysaccharides product.
Preferably, in some embodiments of the invention, the step 2) pot group type dynamic countercurrent extracting method be specially：
A, it is one group with several tanks, tank group first put into temperature before formal circulation is extracted to the marine algae powder in the first tank
It is 60~80 DEG C of water, the extract solution feeding storage tank of its first time；Thereafter second input temperature is 60~80 DEG C of extract solutions of water
Used as the solvent that the same marine algae powder of the second tank is extracted, the extract solution feeding storage tank of the second tank first time, the second tank is secondary
The solvent that extract solution is extracted as the same marine algae powder of the 3rd tank, the first extract solution feeding storage tank of the 3rd tank；First tank the 3rd
It is secondary input temperature be 60~80 DEG C of water, its extract solution as the same marine algae powder of the second tank extraction solvent, the second tank is for the third time
Input temperature be 60~80 DEG C of water, its extract solution as the 3rd same marine algae powder of tank extraction solvent；
B, tank group in formal circulation extraction process, to repeatedly extracting and be last time extraction tank in marine alga
Powder input temperature is 60~80 DEG C of water, and its extract solution is successively as the solvent that each tank marine algae powder is extracted thereafter.
In the tank that c, last time are extracted after marine alga waste residue is discharged, then same marine algae powder is put into, as next round
Tank group circulates the tail tank of extraction successively, and it puts into the feeding storage tank of the first time extract solution after marine algae powder again；
D, the acquisition after multiple dynamic countercurrent is extracted in extraction process per each extract solution of tank marine algae powder.
Preferably, in some embodiments of the invention, the step 3) ultrasound condition be：Ultrasonic power be 400w~
500w, time 20min~30min.
Preferably, in some embodiments of the invention, the flocculant is that active amino is given birth to the cation of hydroxyl
Thing flocculant ionizes out the anionic flocculant of sulfonate ion when being dissolved in water or is their mixed flocculation agents.
Preferably, in some embodiments of the invention, the cationic flocculant of the active amino and hydroxyl
It is shitosan or modified starch；The anionic flocculant that sulfonate ion is ionized out when being dissolved in water is polystyrolsulfon acid
Sodium, neopelex or sulfomethylation polyacrylamide.
Preferably, in some embodiments of the invention, the step 5) be：Under 1600~2300rpm mixing speeds, plus
Enter flocculant, and a pair of plates formula electrode is set in stirred reactor, the two poles end of plate electrode accesses direct current or exchange
Power supply, flocculation of electric field extraction is carried out under the conditions of electric-field intensity is 20~50V/cm to the extraction feed liquid containing algal polysaccharides, flocculation
The addition of agent is 600~900ppm, and its stirring and flocculation extraction time are 20~30min.
Preferably, in some embodiments of the invention, the step 7) purification process purified for radial flow chromatogram.
Beneficial effects of the present invention：
1st, by Mechanical Crushing combination cyclic ultrasonic breaking, effectively cell can be crushed, inside cell
Polysaccharide is discharged.
2nd, by pot group type dynamic countercurrent extracting method can effectively by the Polyose extraction inside alginic cell out, should
The more traditional heating extraction of mode, extraction recovery is higher.
3rd, in order to preferably improve polysaccharide recovery, marine alga waste residue is carried out into ultrasonic assistant extraction, will be residual in waste residue
Staying polysaccharide carries out second extraction, improves the rate of recovery.
4th, flocculant is added, flocculation extraction is carried out in the case of electric field or non-electric field, and with flocculation of electric field extraction effect
More preferably, the formation effect of flocculation core is only played in the addition of its flocculant, and the flocculation of algal polysaccharides is separated out mainly by electric field action
The micro-current provided with negative electrode, thus flocculant usage is only hundreds of ppm (in flocculation feed liquid), flocculant cost is only that ethanol sinks
, to 1/tens, so as to significantly reduce algal polysaccharides extraction cost, and product purity is high for the part in shallow lake.
Specific embodiment
Embodiment 1
Pot group type dynamic countercurrent extracting method is specially：
A, it is one group with several tanks, tank group first put into temperature before formal circulation is extracted to the marine algae powder in the first tank
It is 65 DEG C of water, the extract solution feeding storage tank of its first time；Thereafter input temperature is 65 DEG C of extract solutions of water as second for the second time
The solvent that the same marine algae powder of tank is extracted, the extract solution feeding storage tank of the second tank first time, the secondary extract solution of the second tank is made
It is the solvent that the same marine algae powder of the 3rd tank is extracted, the first extract solution feeding storage tank of the 3rd tank；First tank third time puts into temperature
It is 65 DEG C of water to spend, its extract solution as the same marine algae powder of the second tank extraction solvent, the second tank third time input temperature be 65
DEG C water, its extract solution as the 3rd same marine algae powder of tank extraction solvent；
B, tank group in formal circulation extraction process, to repeatedly extracting and be last time extraction tank in marine alga
Powder input temperature is 65 DEG C of water, and its extract solution is successively as the solvent that each tank marine algae powder is extracted thereafter.
In the tank that c, last time are extracted after marine alga waste residue is discharged, then same marine algae powder is put into, as next round
Tank group circulates the tail tank of extraction successively, and it puts into the feeding storage tank of the first time extract solution after marine algae powder again；
D, the acquisition after multiple dynamic countercurrent is extracted in extraction process per each extract solution of tank marine algae powder.
Embodiment 2
Pot group type dynamic countercurrent extracting method is substantially the same manner as Example 1, and difference is that the temperature of water is 80 DEG C.
Embodiment 3
A kind of preparation method of algal polysaccharides extract, comprises the following steps：
1) dried after marine alga is cleaned, through with mechanical crushing, then ensure that alginic cell is broken completely with cyclic ultrasonic breaking
It is bad, obtain marine algae powder；
2) by step 1) powder be put into and stir in extractor to enter using pot group type dynamic countercurrent extracting method (embodiment 1)
Row is extracted, and obtains extract solution and marine alga waste residue；
3) by step 2) marine alga waste residue be put into 60 DEG C of hot water, carry out ultrasonic wave extraction, ultrasonic power is 400w, time
30min, obtains leaching liquor；
4) by step 2) obtain extract solution and step 3) leaching liquor insert in stirring reaction container, adjust stirring reaction
Extracting temperature is 40 DEG C in device, and pH value is 7；
5) under 1900rpm mixing speeds, to step 4) stirring reaction container in add neopelex wadding
Solidifying agent, and a pair of plates formula electrode is set in stirred reactor, the two poles end of plate electrode accesses direct current or AC power,
Flocculation of electric field extraction, the addition of flocculant are carried out to the extraction feed liquid containing algal polysaccharides under the conditions of electric-field intensity is 30V/cm
It is 700ppm, its stirring and flocculation extraction time are 24min；Its flocculation recovery rate is 72.6%.
Embodiment 4
A kind of preparation method of algal polysaccharides extract, comprises the following steps：
1) dried after marine alga is cleaned, through with mechanical crushing, then ensure that alginic cell is broken completely with cyclic ultrasonic breaking
It is bad, obtain marine algae powder；
2) by step 1) powder be put into and stir in extractor to enter using pot group type dynamic countercurrent extracting method (embodiment 2)
Row is extracted, and obtains extract solution and marine alga waste residue；
3) by step 2) marine alga waste residue be put into 80 DEG C of hot water, carry out ultrasonic wave extraction, ultrasonic power is 450w, time
26min, obtains leaching liquor；
4) by step 2) obtain extract solution and step 3) leaching liquor insert in stirring reaction container, adjust stirring reaction
Extracting temperature is 30 DEG C in device, and pH value is 6；
5) under 1600rpm mixing speeds, to step 4) stirring reaction container in add neopelex wadding
Solidifying agent, and a pair of plates formula electrode is set in stirred reactor, the two poles end of plate electrode accesses direct current or AC power,
Flocculation of electric field extraction, the addition of flocculant are carried out to the extraction feed liquid containing algal polysaccharides under the conditions of electric-field intensity is 20V/cm
It is 600ppm, its stirring and flocculation extraction time are 30min；Its flocculation recovery rate is 68.1%.
Embodiment 5
A kind of preparation method of algal polysaccharides extract, comprises the following steps：
1) dried after marine alga is cleaned, through with mechanical crushing, then ensure that alginic cell is broken completely with cyclic ultrasonic breaking
It is bad, obtain marine algae powder；
2) by step 1) powder be put into extractor to stir and extracted using pot group type dynamic countercurrent extracting method, obtain
To extract solution and marine alga waste residue；
3) by step 2) marine alga waste residue be put into 100 DEG C of hot water, carry out ultrasonic wave extraction, obtain leaching liquor；
4) by step 2) obtain extract solution and step 3) leaching liquor insert in stirring reaction container, adjust stirring reaction
Extracting temperature is 60 DEG C in device, and pH value is 8；
5) under 1600~2300rpm mixing speeds, to step 4) stirring reaction container in add polystyrolsulfon acid
Sodium flocculant, and a pair of plates formula electrode is set in stirred reactor, the two poles end of plate electrode accesses direct current or exchange
Power supply, flocculation of electric field extraction is carried out under the conditions of electric-field intensity is 50V/cm to the extraction feed liquid containing algal polysaccharides, flocculant
Addition is 900ppm, and its stirring and flocculation extraction time are 20min.Its flocculation recovery rate is 70.5%.
Embodiment 6
Exist according to Min Yutao, Song Yanxian etc. 2015《Food industry》Entitled " the algal polysaccharides that the 6th phase of volume 36 delivers
Two kinds of extracting methods and its hypoglycemic activity compare ", carry out hypoglycemic activity experiment.It is many with the marine alga that embodiment 1-3 is prepared
Sugared crude product is tested, experimental result：Embodiment 1-3 the inhibiting rate of amylopsin is respectively 91.3%, 89.2% and
90.5%.
Presently preferred embodiments of the present invention is the foregoing is only, is not intended to limit the invention, it is all in essence of the invention
Within god and principle, any modification, equivalent substitution and improvements made etc. should be included within the scope of the present invention.
Claims (7)
1. a kind of preparation method of algal polysaccharides extract, it is characterised in that comprise the following steps：
1) dried after marine alga is cleaned, through with mechanical crushing, then ensure that alginic cell is totally disrupted with cyclic ultrasonic breaking, obtained
To marine algae powder；
2) by step 1) powder be put into extractor to stir and extracted using pot group type dynamic countercurrent extracting method, carried
Take liquid and marine alga waste residue；
3) by step 2) marine alga waste residue be put into 60~100 DEG C of hot water, carry out ultrasonic wave extraction, obtain leaching liquor；
4) by step 2) obtain extract solution and step 3) leaching liquor insert in stirring reaction container, regulation stirred reactor in
Extracting temperature is 20~60 DEG C, and pH value is 5~8；
5) to step 4) stirring reaction container in add 600~900ppm flocculant, its stirring and flocculation extraction time be
20~30min；
6) after the completion of flocculation is extracted, flocculation extract solution is clarified or centrifugal filtration, is obtained algal polysaccharides crude product；
7) the algal polysaccharides crude product that will be obtained is further purified, and obtains algal polysaccharides product.
2. the preparation method of algal polysaccharides extract according to claim 1, it is characterised in that the step 2) tank group
Formula dynamic countercurrent extracting method is specially：
A, it is one group with several tanks, tank group is first 60 to the marine algae powder input temperature in the first tank before formal circulation is extracted
~80 DEG C of water, the extract solution feeding storage tank of its first time；Thereafter second input temperature is 60~80 DEG C of extract solution conducts of water
The solvent that the same marine algae powder of second tank is extracted, the extract solution feeding storage tank of the second tank first time, the secondary extraction of the second tank
The solvent that liquid is extracted as the same marine algae powder of the 3rd tank, the first extract solution feeding storage tank of the 3rd tank；First tank third time is thrown
Enter temperature for 60~80 DEG C of water, its extract solution as the same marine algae powder of the second tank extraction solvent, the second tank third time puts into
Temperature be 60~80 DEG C of water, its extract solution as the 3rd same marine algae powder of tank extraction solvent；
B, tank group in formal circulation extraction process, to repeatedly extracting and be last time extraction tank in marine algae powder
Input temperature is 60~80 DEG C of water, and its extract solution is successively as the solvent that each tank marine algae powder is extracted thereafter.
In the tank that c, last time are extracted after marine alga waste residue is discharged, then same marine algae powder is put into, as next round tank group
The tail tank of extraction is circulated successively, and it puts into the feeding storage tank of the first time extract solution after marine algae powder again；
D, the acquisition after multiple dynamic countercurrent is extracted in extraction process per each extract solution of tank marine algae powder.
3. the preparation method of algal polysaccharides extract according to claim 1, it is characterised in that the step 3) ultrasound
Condition is：Ultrasonic power is 400w~500w, time 20min~30min.
4. the preparation method of algal polysaccharides extract according to claim 1, it is characterised in that the flocculant be with
The cationic flocculant of active amino and hydroxyl or ionize out when being dissolved in water sulfonate ion anionic flocculant or
It is their mixed flocculation agents.
5. the preparation method of algal polysaccharides extract according to claim 4, it is characterised in that the active amino
It is shitosan or modified starch with the cationic flocculant of hydroxyl；It is described ionize out when being dissolved in water sulfonate ion it is cloudy from
Subtype flocculant is kayexalate, neopelex or sulfomethylation polyacrylamide.
6. the preparation method of algal polysaccharides extract according to claim 1, it is characterised in that the step 5) be：
Under 1600~2300rpm mixing speeds, flocculant is added, and a pair of plates formula electrode is set in stirred reactor, it is flat
The two poles end of electrode accesses direct current or AC power, to the extraction containing algal polysaccharides under the conditions of electric-field intensity is 20~50V/cm
Feed liquid carries out flocculation of electric field extraction, and the addition of flocculant is 600~900ppm, and its stirring and flocculation extraction time be 20~
30min。
7. the preparation method of algal polysaccharides extract according to claim 1, it is characterised in that the step 7) purifying
Method is purified for radial flow chromatogram.
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| CN107625718A (en) * | 2017-09-11 | 2018-01-26 | 浙江海洋大学 | A kind of compound algal polysaccharides facial mask liquid |
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| CN107641160A (en) * | 2017-10-31 | 2018-01-30 | 青岛绿浒苔农业科技有限公司 | Utilize the method for multi-stage countercurrent enzymolysis coupling technology extraction sea grass polysaccharide |
| CN107998156A (en) * | 2017-12-14 | 2018-05-08 | 浦江县昂宝生物技术有限公司 | The preparation method of anti-thrombotic agents |
| CN108042580A (en) * | 2017-12-14 | 2018-05-18 | 金华市艾力生物科技有限公司 | The preparation method of marine algae extract with reducing blood lipid effectiveness |
| CN110183544A (en) * | 2019-06-06 | 2019-08-30 | 山东天易科技有限公司 | A kind of preparation method of algal polysaccharides |
| CN110236172A (en) * | 2019-07-25 | 2019-09-17 | 孙杨 | A kind of food seaweed algae salt production process |
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