CN111393488A - Method for purifying and concentrating water-soluble chitosan oligosaccharide - Google Patents
Method for purifying and concentrating water-soluble chitosan oligosaccharide Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/04—Disaccharides
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
- C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
- C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
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- Genetics & Genomics (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
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- Sustainable Development (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention relates to a method for purifying and concentrating water-soluble chitosan oligosaccharide, which comprises the following steps: (1) pretreating the chitosan oligosaccharide degradation solution through a ceramic membrane to remove chitosan oligosaccharide and other insoluble impurities in the degradation solution to obtain ceramic membrane permeate; (2) purifying the ceramic membrane permeate by using an ultrafiltration membrane to remove macromolecular polysaccharide and other impurities to obtain ultrafiltration permeate; (3) concentrating the ultrafiltration permeate by using a nanofiltration membrane, adding water, performing four-stage separation and concentration, and removing inorganic salt and monosaccharide to obtain a chitosan oligosaccharide nanofiltration concentrate; (4) and (3) carrying out spray drying on the finally concentrated chitosan oligosaccharide concentrated solution to obtain high-purity chitosan oligosaccharide powder. The invention has simple process, no phase change in the process, low energy consumption and easy industrial production. The method can improve the yield of chitosan oligosaccharide, obtain high-purity chitobiose-chitohexaose with average molecular weight less than 1000, and the obtained chitosan oligosaccharide has high purity and narrow molecular weight distribution, and is suitable for preparing chitosan oligosaccharide products conforming to pharmaceutical grade.
Description
Technical Field
The invention relates to the technical field of chitosan oligosaccharide purification and concentration, in particular to a method for purifying and concentrating water-soluble chitosan oligosaccharide.
Technical Field
The chitosan oligosaccharide is called chitosan oligosaccharide and oligochitosan, is an oligosaccharide product with the polymerization degree of 2-20, is obtained by degrading deacetylated products of shrimp and crab shells by a special biological enzyme technology (reports of using chemical degradation and microwave degradation technologies), has the molecular weight of less than or equal to 3200Da, and is a low-molecular-weight product with good water solubility, large functional effect and high biological activity. It has several unique functions of high solubility, complete water solubility, easy absorption and utilization by organism, etc. and its action is 14 times that of chitosan.
The chitosan oligosaccharide is the only cationic basic amino oligosaccharide with positive charges in the nature, and is animal cellulose. Chitosan oligosaccharide is small molecular oligosaccharide with amino group degraded by chitosan derived from shrimp and crab shell, and is sugar chain with polymerization degree of 2-20.
The processing of the high molecular chitosan into the water-soluble low molecular chitosan oligosaccharide by a microwave physical method is a new biotechnology following genetic engineering and protein engineering, is called as a third-generation biotechnology, and can be widely applied to the fields of agriculture, food, chemical industry, energy, environmental protection, medicine and the like. The following is the structural formula of chitosan oligosaccharide:
the main methods for degrading chitosan include acidolysis and enzymolysis. The acidolysis method utilizes the characteristic that a plurality of free amino groups in chitosan molecules can be combined with hydrogen ions in a solution to cause the hydrogen bonds between the chitosan molecules and the inside of the chitosan molecules to be broken, so that the molecular structure is stretched, and the long-chain part is easy to break sugar-shake bonds to form a plurality of molecular fragments with different polymerization degrees. Compared with other degradation methods, the enzymatic (degradation) method has the advantages of mild reaction conditions, easily controlled degradation process and relative molecular weight distribution of degradation products, high bioactivity of the prepared chitosan oligosaccharide, no need of desalting the product, easily controlled process, no pollution to the environment and the like, and is an ideal degradation method.
Generally, chitosan is directly subjected to spray drying after being degraded by a biological enzyme method in industry, and the obtained chitosan oligosaccharide product has generally wide molecular weight distribution, contains some chitosan macromolecules which are not fully degraded, chitosan degrading enzymes, inorganic salts, acid radical ions, saccharides with low biological activity such as monosaccharide and disaccharide, and other impurity components in raw material chitosan, and the like, and the components can influence the purity of chitosan oligosaccharide and further influence the use value of the chitosan oligosaccharide. In order to obtain a chitosan oligosaccharide product with high chitotriose-chitohexaose content, a proper separation and purification means is required to be selected to purify the enzymolysis liquid.
The chitosan oligosaccharide molecule contains more amino and hydroxyl groups, so that the intermolecular or intramolecular action is strong, and the separation and purification are relatively difficult. At present, the separation and purification methods of chitosan oligosaccharide mainly comprise: membrane separation, gel permeation chromatography, thin layer chromatography, ion exchange chromatography, and the like.
The membrane separation technology is a new separation technology which rises rapidly after the 60's of the 20 th century. The technology has the functions of separation, concentration, purification and refining, and has the characteristics of high efficiency, energy conservation, environmental protection, simple molecular-level filtration and filtration process, easy control and the like, so the technology is widely applied to the fields of food, medicine, biology, environmental protection, chemical industry, metallurgy, energy, petroleum, water treatment, electronics, bionics and the like, generates great economic benefit and social benefit, and becomes one of the most important means in the current separation science.
CN103073600A discloses a method for preparing water-soluble chitosan oligosaccharide, which comprises the steps of carrying out continuous microwave treatment on high-molecular chitosan, separating by a nanofiltration membrane after degradation treatment, concentrating by a centrifugal machine, drying in vacuum at low temperature, and crushing to obtain the chitosan oligosaccharide finished product. The method needs complicated vacuum-pumping drying, the post-treatment process is complicated, the energy consumption of centrifugal machine centrifugation, vacuum drying and the like is high, the production cost is overhigh, and the purity of the obtained chitosan oligosaccharide is lower.
CN102120780A discloses a method for preparing chitosan oligosaccharide by semi-wet microwave treatment, which takes chitosan colloidal powder as a raw material and prepares the chitosan oligosaccharide through the steps of hydration, microwave treatment, water-soluble neutralization, membrane separation, drying and the like. The method has complex hydration process, needs acid and alkali as raw materials for reaction, has higher production cost and lower purity of the obtained chitosan oligosaccharide.
CN103641936B discloses a method for preparing water-soluble chitosan oligosaccharide by ultrafiltration and nanofiltration, which separates active oligosaccharide concentrated solution by ultrafiltration and nanofiltration separation technology, but the molecular weight of the chitosan oligosaccharide obtained by the method is between 300 and 25000, the molecular weight distribution is wider, and the yield and the purity are relatively lower for the chitosan oligosaccharide with higher requirements such as pharmaceutical grade (the chitosan oligosaccharide needs high purity and narrow molecular weight distribution).
Disclosure of Invention
The invention aims to provide a method for purifying and concentrating water-soluble chitosan oligosaccharide, which has the advantages of no phase change in the process, low energy consumption, easy industrial production, high chitosan oligosaccharide yield improvement, high purity of the obtained chitosan oligosaccharide, narrow molecular weight distribution, suitability for preparing a chitosan oligosaccharide product conforming to the pharmaceutical grade and improvement of the added value of the chitosan oligosaccharide product.
In order to achieve the above object, the present invention provides a method for purifying and concentrating water-soluble chitosan oligosaccharide, comprising the following steps:
(1) pretreating the chitosan oligosaccharide degradation solution through a ceramic membrane to remove insoluble impurities (including chitosan oligosaccharide and other insoluble impurities) in the degradation solution to obtain ceramic membrane permeation solution;
(2) purifying the ceramic membrane permeate by using an ultrafiltration membrane to remove macromolecular proteins and macromolecular polysaccharides to obtain ultrafiltration permeate;
(3) concentrating the ultrafiltration permeate by using a nanofiltration membrane, adding water, performing four-stage separation and concentration, and removing inorganic salt and monosaccharide to obtain a chitosan oligosaccharide nanofiltration concentrate;
(4) spray drying the finally concentrated chitosan oligosaccharide concentrated solution to obtain high-purity chitosan oligosaccharide powder;
in the step (3), the concentration proportion of each stage of four-stage separation and concentration by adopting a nanofiltration membrane is 2-5 times, and the water adding proportion of each stage is 1-4 times.
Further, in the step (3), the used nanofiltration membrane is one of a hollow fiber nanofiltration membrane, a flat plate nanofiltration membrane, a roll nanofiltration membrane and a disc tube nanofiltration membrane; the nanofiltration membrane material is one or more organic materials of polyamide, polysulfone, polyethersulfone and the like, polyimide, polyacrylonitrile, hyperbranched polymer and polyethyleneimine or one or more inorganic materials of alumina, zirconia, titanium dioxide, silicon carbide and graphene or organic-inorganic hybrid materials; the cut-off molecular weight of the nanofiltration membrane is 150-800 Da.
Further, in the step (1), the ceramic membrane used is one of a tubular ceramic membrane, a flat plate ceramic membrane and a multi-channel ceramic membrane; the ceramic membrane material is one or more of inorganic materials such as alumina, zirconia, titanium oxide, silicon carbide and the like; the aperture of the ceramic membrane is 20-200 nm.
Further, in the step (2), the ultrafiltration membrane used is one of a hollow fiber, a flat plate, a roll-type ultrafiltration membrane and a tubular ultrafiltration membrane; the used ultrafiltration membrane material is one or more inorganic materials of ceramics, glass, alumina, zirconia and metal or one or more organic materials of polypropylene, polyvinylidene fluoride, polytetrafluoroethylene, polysulfone and polyether sulfone; the cutoff molecular weight of the used ultrafiltration membrane is 5000-20000 Da.
And (3) recycling the third-stage nanofiltration membrane permeate and the fourth-stage nanofiltration membrane permeate to the chitosan oligosaccharide degradation production.
And (3) performing four-stage separation and concentration by using nanofiltration membranes, wherein the nanofiltration membranes of different materials and specifications are used for fractional separation and concentration respectively.
Further, in the step (1), the chitosan oligosaccharide degradation solution is a chitosan oligosaccharide degradation solution prepared by a chemical preparation method or a physical preparation method.
Still further, the chemical preparation method is an acidolysis method or an oxidative degradation method.
Further, the physical preparation method is an enzymolysis method, a microwave method or a composite degradation method.
The invention has the technical effects that: the invention has simple process, no phase change in the process, low energy consumption and easy industrial production. The method can improve the yield of chitosan oligosaccharide, and obtain high-purity and high-added-value chitosan oligosaccharide product. Specifically, the chitosan-chitohexaose with high purity and average molecular weight less than 1000 is obtained, the molecular weight distribution is narrow, the molecular weight of the chitosan oligosaccharide is between 600 and 1500, and the obtained chitosan oligosaccharide has high purity and narrow molecular weight distribution, is suitable for preparing chitosan oligosaccharide products conforming to the pharmaceutical grade, and improves the added value of the chitosan oligosaccharide products.
Drawings
FIG. 1 is a process flow diagram of the present invention;
FIG. 2 is a process flow chart of the present invention in which the concentration ratio of each stage is 5 times and the water addition ratio is 4 times;
FIG. 3 is a process flow chart of the present invention in which the concentration ratio of each stage is 4 times and the water addition ratio is 3 times.
Detailed Description
The invention relates to a method for purifying and concentrating water-soluble chitosan oligosaccharide, which comprises the following steps: (1) pretreating the chitosan oligosaccharide degradation solution through a ceramic membrane to remove chitosan oligosaccharide and other insoluble impurities in the degradation solution to obtain ceramic membrane permeate; (2) purifying the ceramic membrane permeate by using an ultrafiltration membrane to remove macromolecular polysaccharide and other impurities to obtain ultrafiltration permeate; (3) concentrating the ultrafiltration permeate by using a nanofiltration membrane, adding water, performing four-stage separation and concentration, and removing inorganic salt and monosaccharide to obtain a chitosan oligosaccharide nanofiltration concentrate; (4) and (3) carrying out spray drying on the finally concentrated chitosan oligosaccharide concentrated solution to obtain high-purity chitosan oligosaccharide powder. The invention has simple process, no phase change in the process, low energy consumption and easy industrial production. The method can improve the yield of chitosan oligosaccharide, obtain high-purity chitobiose-chitohexaose with average molecular weight less than 1000, and the obtained chitosan oligosaccharide has high purity and narrow molecular weight distribution, and is suitable for preparing chitosan oligosaccharide products conforming to pharmaceutical grade, and the additional value of the chitosan oligosaccharide products is improved.
Specifically, the method for purifying and concentrating the water-soluble chitosan oligosaccharide comprises the following steps:
(1) pretreating the chitosan oligosaccharide degradation solution through a ceramic membrane to remove chitosan oligosaccharide and other insoluble impurities in the degradation solution to obtain ceramic membrane permeate;
(2) purifying the ceramic membrane permeate by using an ultrafiltration membrane to remove macromolecular proteins and macromolecular polysaccharides to obtain ultrafiltration permeate;
(3) concentrating the ultrafiltration permeate by using a nanofiltration membrane, adding water, performing four-stage separation and concentration (namely concentration, adding water, concentration, adding water and concentration), and removing inorganic salt and monosaccharide to obtain a chitosan oligosaccharide nanofiltration concentrate;
(4) and (3) carrying out spray drying on the finally concentrated chitosan oligosaccharide concentrated solution to obtain high-purity chitosan oligosaccharide powder.
Further, in the step (1), the chitosan oligosaccharide degradation solution may be any one of a chemical preparation method (acid hydrolysis, oxidative degradation, etc.) and a physical preparation method (enzymatic hydrolysis, microwave method, composite degradation method, etc.).
Further, in the step (1), the ceramic membrane used is one of a tubular type, a flat plate type and a multi-channel ceramic membrane.
Still further, in the step (1), the ceramic film material used is one or a composite material of two or more of inorganic materials such as alumina, zirconia, titania, silicon carbide, and the like.
Further, in the step (1), the pore diameter of the ceramic film used is 20 to 200 nm.
Further, in the step (2), the ultrafiltration membrane used is one of a hollow fiber, a flat plate, a roll-type ultrafiltration membrane and a tubular ultrafiltration membrane.
Still further, in the step (2), the ultrafiltration membrane material used is one of inorganic (further, ceramic, glass, alumina, zirconia, metal, etc.) and organic (further, polypropylene, polyvinylidene fluoride, polytetrafluoroethylene, polysulfone, polyethersulfone, etc.) materials.
Further, in the step (2), the cut-off molecular weight of the ultrafiltration membrane used is 5000-20000 Da.
Further, in the step (3), the nanofiltration membrane is one of a hollow fiber nanofiltration membrane, a flat plate nanofiltration membrane, a roll nanofiltration membrane and a disc tube nanofiltration membrane.
Still further, in the step (3), the nanofiltration membrane material used is one of an organic material (further, polyamide, polysulfone, polyethersulfone, polyimide, polyacrylonitrile, hyperbranched polymer, polyethyleneimine, and the like), an inorganic material (further, alumina, zirconia, titanium dioxide, silicon carbide, graphene, and the like) and an organic-inorganic hybrid material.
Furthermore, in the step (3), the cut-off molecular weight of the nanofiltration membrane is 150-800 Da.
Further, in the step (3), a nanofiltration membrane is adopted for four-stage separation (and) concentration, the concentration proportion of each stage is 2-5 times, and the water adding proportion of each stage is 1-4 times.
That is, referring to fig. 2, the concentration ratio of each stage is 5 times, and the water addition ratio of each stage is 4 times.
Referring to fig. 3, the concentration ratio of each stage is 4 times, and the water addition ratio of each stage is 3 times.
Furthermore, in the step (3), the three-stage nanofiltration membrane permeate and the four-stage nanofiltration membrane permeate (namely, the permeate obtained after concentration, water addition, concentration, water addition and concentration, and finally water addition in two stages) can be reused in the production of chitosan oligosaccharide degradation. The two-stage nanofiltration membrane permeate liquid has high quality, can be recycled, and further saves the production cost.
Still further, in the step (3), the nanofiltration membrane is used for four-stage separation and concentration, and the nanofiltration membranes with different materials and specifications (specifications of cut-off molecular weight of the nanofiltration membrane) can be used for fractional separation and concentration respectively.
The invention is further illustrated by the following two examples.
The first embodiment is as follows:
the embodiment comprises the following steps:
1. a method for purifying and concentrating water-soluble chitosan oligosaccharide comprises the following steps:
(1) filtering and pretreating 100L chitosan oligosaccharide degradation solution (chitosan content is about 3.0%) through a porous alumina ceramic membrane with pore diameter of 50nm to remove undegraded chitosan and other insoluble impurities to obtain 95.0L permeate;
(2) purifying the ceramic membrane permeate by adopting a polysulfone roll-type ultrafiltration membrane with the molecular weight cutoff of 8000Da to remove macromolecular polysaccharide and other impurities to obtain 90.0L ultrafiltration permeate;
(3) concentrating the ultrafiltration permeate by using a polyamide roll-type nanofiltration membrane with the molecular weight cutoff of 500Da, adding water, performing four-stage separation and concentration (namely concentration, adding water, concentration, adding water and concentration), wherein the concentration ratio is 3 times, the water addition ratio is 2 times, and removing inorganic salt and monosaccharide to finally obtain 30.0L chitosan oligosaccharide nanofiltration concentrate;
(4) spray drying the final concentrated chitosan oligosaccharide concentrate to obtain 2.21kg of high-purity chitosan oligosaccharide powder with the yield of 70.7%.
(5) Sampling, detecting and analyzing, wherein the detection and analysis results are as follows:
example two:
the embodiment comprises the following steps:
1. a method for purifying and concentrating water-soluble chitosan oligosaccharide comprises the following steps:
(1) filtering and pretreating 200L chitosan oligosaccharide degradation liquid (chitosan content is about 2.5%) by a plate-type silicon carbide ceramic membrane with the pore diameter of 200nm to remove undegraded chitosan and other insoluble impurities to obtain 192.0L permeate;
(2) purifying the ceramic membrane permeate by adopting a polyethersulfone roll-type ultrafiltration membrane with the molecular weight cutoff of 5000Da to remove macromolecular polysaccharide and other impurities to obtain 180.0L ultrafiltration permeate;
(3) carrying out primary and secondary concentration-water separation and concentration (concentration-water addition-concentration) on the ultrafiltration permeate by adopting a polyamide roll-type nanofiltration membrane with the molecular weight cutoff of 500Da, carrying out tertiary and quaternary concentration-water separation and concentration (water addition-concentration-water addition-concentration) on the ultrafiltration permeate by adopting a polyether sulfone roll-type nanofiltration membrane with the molecular weight cutoff of 200Da, wherein the concentration ratio is 4 times, the water addition ratio is 3 times, and inorganic salt and monosaccharide are removed to finally obtain 45.0L chitosan oligosaccharide nanofiltration concentrate;
(4) the final concentrated chitosan oligosaccharide concentrate was spray dried to obtain 3.86kg of high purity chitosan oligosaccharide powder with a yield of 77.2%.
(5) Sampling, detecting and analyzing, wherein the detection and analysis results are as follows:
it will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (9)
1. A method for purifying and concentrating water-soluble chitosan oligosaccharide is characterized by comprising the following steps:
(1) pretreating the chitosan oligosaccharide degradation solution through a ceramic membrane to remove insoluble impurities in the degradation solution to obtain ceramic membrane permeation solution;
(2) purifying the ceramic membrane permeate by using an ultrafiltration membrane to remove macromolecular proteins and macromolecular polysaccharides to obtain ultrafiltration permeate;
(3) concentrating the ultrafiltration permeate by using a nanofiltration membrane, adding water, performing four-stage separation and concentration, and removing inorganic salt and monosaccharide to obtain a chitosan oligosaccharide nanofiltration concentrate;
(4) spray drying the finally concentrated chitosan oligosaccharide concentrated solution to obtain high-purity chitosan oligosaccharide powder;
in the step (3), the concentration proportion of each stage of the four-stage separation and concentration by adopting the nanofiltration membrane is 2-5 times, and the water adding proportion of each stage is 1-4 times.
2. The method for purifying and concentrating water-soluble chitosan oligosaccharide as claimed in claim 1, wherein: in the step (3), the used nanofiltration membrane is one of a hollow fiber nanofiltration membrane, a flat plate nanofiltration membrane, a roll nanofiltration membrane and a disc tube nanofiltration membrane; the nanofiltration membrane material is one or more organic materials of polyamide, polysulfone, polyethersulfone and the like, polyimide, polyacrylonitrile, hyperbranched polymer and polyethyleneimine or one or more inorganic materials of alumina, zirconia, titanium dioxide, silicon carbide and graphene or organic-inorganic hybrid materials; the cut-off molecular weight of the nanofiltration membrane is 150-800 Da.
3. The method for purifying and concentrating water-soluble chitosan oligosaccharide as claimed in claim 1, wherein: in the step (1), the ceramic membrane is one of a tubular ceramic membrane, a flat plate ceramic membrane and a multi-channel ceramic membrane; the ceramic membrane material is one or more of inorganic materials such as alumina, zirconia, titanium oxide, silicon carbide and the like; the aperture of the ceramic membrane is 20-200 nm.
4. The method for purifying and concentrating water-soluble chitosan oligosaccharide as claimed in claim 1, wherein: in the step (2), the ultrafiltration membrane is one of a hollow fiber, a flat plate, a roll-type ultrafiltration membrane and a tubular ultrafiltration membrane; the used ultrafiltration membrane material is one or more inorganic materials of ceramics, glass, alumina, zirconia and metal or one or more organic materials of polypropylene, polyvinylidene fluoride, polytetrafluoroethylene, polysulfone and polyether sulfone; the cutoff molecular weight of the used ultrafiltration membrane is 5000-20000 Da.
5. The method for purifying and concentrating water-soluble chitosan oligosaccharide as claimed in claim 1, wherein: and (4) in the step (3), the three-stage nanofiltration membrane permeate and the four-stage nanofiltration membrane permeate are recycled in the production of chitosan oligosaccharide degradation.
6. The method for purifying and concentrating water-soluble chitosan oligosaccharide as claimed in claim 2, wherein: and (3) performing four-stage separation and concentration by adopting nanofiltration membranes, wherein the nanofiltration membranes of different materials and specifications are used for performing fractional separation and concentration respectively.
7. The method for purifying and concentrating water-soluble chitosan oligosaccharide as claimed in claim 1, wherein: in the step (1), the chitosan oligosaccharide degradation solution is prepared by a chemical preparation method or a physical preparation method.
8. The method for purifying and concentrating water-soluble chitosan oligosaccharide as claimed in claim 7, wherein: the chemical preparation method is an acidolysis method or an oxidative degradation method.
9. The method for purifying and concentrating water-soluble chitosan oligosaccharide as claimed in claim 7, wherein: the physical preparation method is an enzymolysis method, a microwave method or a composite degradation method.
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