CN110819743A - Preparation method of low-sensitization fishy smell-removed antarctic ice algae oligosaccharide - Google Patents
Preparation method of low-sensitization fishy smell-removed antarctic ice algae oligosaccharide Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C13—SUGAR INDUSTRY
- C13K—SACCHARIDES OBTAINED FROM NATURAL SOURCES OR BY HYDROLYSIS OF NATURALLY OCCURRING DISACCHARIDES, OLIGOSACCHARIDES OR POLYSACCHARIDES
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Abstract
The invention relates to a preparation method of low-sensitization fishy smell-removed antarctic ice algae oligosaccharide, which sequentially comprises the steps of crude oligosaccharide preparation, ultrafiltration, flash evaporation and membrane classification according to the preparation sequence; the membrane fractionation is to separate the concentrated oligose liquid obtained by flash evaporation into a plurality of components with different molecular weights by using a semi-permeable membrane device, detect the allergen content of each component except the component with the minimum molecular weight, discard the component with the highest allergen content and the component with the minimum molecular weight, and collect and combine other components. The invention effectively removes the sensitizing substances in the Antarctic ice algae oligosaccharide by a multilayer directional dialysis technology, and simultaneously adopts a one-step concentration method of ultrafiltration flash evaporation series connection to obtain the Antarctic ice algae oligosaccharide which has the characteristics of low sensitization, no fishy smell and the like. The invention has high efficiency of the manufacturing process, easy industrialization, high oligosaccharide activity and obvious effect, simultaneously desensitizes and removes fishy smell, expands the application range of the ice algae oligosaccharide and can be widely applied to the fields of nourishment, skin care products, medicines and the like.
Description
Technical Field
The invention particularly relates to a preparation method of low-sensitization fishy smell-removed antarctic ice algae oligosaccharide.
Background
Antarctic ice algae mainly refers to the large class of microalgae that grow in Antarctic extreme environments in sea ice. The reasonable development of the low-temperature algae resource of the Antarctic can not only enrich the existing microorganism resource of China in application, develop valuable physiologically active substances and products therefrom, but also provide novel research materials and new ideas for basic research. The presently developed Antarctic ice algae comprise two classes, Haematococcus (Durvillaea antarctica) and Crypthecodinium strium (Ultricularia amethystina). The sea antler is a deep-sea brown alga; porphyridium is usually fixed to the sea floor or some solid structure and is a single plant or a long string of simple plants composed of basal cells. Both algae grow annually in uncontaminated waters in the south Pole circle with an average water temperature of no more than 5 ℃. The extreme growth conditions endow the product with pure and rich nutrient substances, the product is rich in a plurality of functional polysaccharides such as fucose, glucan, seaweed gel and the like, has remarkable biological activities such as immunoregulation, antioxidation, blood sugar reduction and the like, and has great application potential in the fields of cosmetics, nutritional foods and medicines.
However, allergic substances are produced during the preparation of antarctic ice algae oligosaccharides; the auxiliary agent added in the preparation is easy to introduce allergic substances, and symptoms such as pruritus, rash, red swelling and the like generally appear. Allergic substances in Antarctic ice algae polysaccharide are various, the system is complex, and the allergic substances are difficult to remove by a common purification method. In addition, the fishy smell characteristic of marine organisms also affects their applications.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a preparation and purification method of low-sensitization deodorization Antarctic krill peptide, which has the advantages of simple process, easy operation, low cost and high desensitization efficiency.
The specific technical scheme is as follows:
a preparation method of low-sensitization deodorization Antarctic ice algae oligosaccharide comprises the steps of crude oligosaccharide preparation, ultrafiltration, flash evaporation and membrane classification in sequence;
the preparation of the crude oligosaccharide takes Antarctic ice algae polysaccharide as a raw material, and the Antarctic ice algae polysaccharide is cracked into oligosaccharide.
The membrane fractionation is to separate the concentrated oligose solution obtained by flash evaporation into a plurality of components with different molecular weights by using a semi-permeable membrane device, detect the allergen content of each component except the component with the minimum molecular weight, discard the component with the highest allergen content and the interval component with the minimum molecular weight, and collect other components. The collected components can be used separately or combined.
The concentrated oligose liquid obtained by flash evaporation is separated into a plurality of components with different molecular weights, namely, the extracting solution is divided into a plurality of molecular weight regions by utilizing a semipermeable membrane.
The Antarctic ice algae is preferably HAIRONG (Durvillaea antarctica) and ZIJINGCAO (Ultriculariaamylothystina).
The inventors have surprisingly found that there is a degree of molecular weight aggregation of the allergenic material in the ice algae oligosaccharide product.
In addition, the small molecular components have more allergic substances, and the small molecular components are removed, so that the small molecular components also have the effects of desalting and removing other small molecular impurities.
The detection method is a thiazole blue method, namely an MTT method for short, the allergen in the ice algae oligosaccharide extracting solution mainly comprises components such as micromolecular protein and micromolecular polar substances, the allergen can penetrate through cell membranes to enter cells, amber dehydrogenase in mitochondria of living cells can enable exogenous MTT to be reduced into blue-purple Formazan (Formazan) which is difficult to dissolve in water, the Formazan is crystallized and deposited in the cells, the crystal can be dissolved by acid isopropanol, an enzyme linked immunosorbent assay instrument is used for measuring the light absorption value at the wavelength of 570nm, the number of the living cells can be indirectly reflected, and the test object ET50 can be detected through an MTT cell activity detection method.
Preferably, the concentrated oligosaccharide extract is separated into 3 to 5 fractions having different molecular weights using a semipermeable membrane device.
The amount of the components is obtained by comprehensively considering desensitization effect, yield and process cost.
More preferably, 4 semipermeable membrane devices are used for separating the concentrated oligose liquid into 5 components with different molecular weights, and the molecular weight cut-off of the 4 semipermeable membrane devices is 700-1600 Da, 500-1200 Da, 300-800 Da and 100-500 Da respectively.
Preferably, the semipermeable membrane device is a dialysis bag.
Dialysis bags with different cut-off molecular weights can be selected for dialysis in sequence from large to small. In particular, the oligosaccharide liquid is dialyzed by a dialysis bag with the largest molecular weight cut-off, the liquid in the bag is reserved, and the dialysis effluent (namely the residual liquid) is dialyzed by a dialysis bag with the next first molecular weight cut-off, and so on.
Further preferably, the semipermeable membrane device comprises a plurality of dialysis bags which are sleeved from inside to outside in sequence according to the molecular weight cut-off from large to small.
The dialysis bags are preferably 4, and the cut-off molecular weights of the dialysis bags are 700-1600 Da, 500-1200 Da, 300-800 Da and 100-500 Da from inside to outside in sequence; the dialysis temperature is 4-15 ℃, and the dialysis time is 12-48 h.
Among them, the most preferable ones are 800Da, 550Da, 350Da, 150Da in turn from inside to outside.
A plurality of dialysis bags which are sleeved from inside to outside in sequence according to the molecular weight cut-off from large to small are used, so that components with a plurality of molecular weights can be obtained simultaneously, multilayer directional dialysis is achieved, and the production process is simplified.
The invention also designs a multilayer directional dialysis device which comprises a dialysis container and a plurality of dialysis bags which are arranged in the container and sleeved from inside to outside in sequence according to the molecular weight cut-off, wherein the diameters of the sections of the dialysis bags are increased from inside to outside in sequence. The multi-layer directional dialysis device can also comprise a plurality of sleeve skeletons corresponding to the shapes and the sizes of the dialysis bags, and the dialysis bags are adhered to the sleeve skeletons so as to fix the shapes of the dialysis bags. When the device is used, concentrated enzymatic hydrolysate obtained by ultrafiltration is poured into the dialysis bag at the innermost layer, a proper amount of water or buffer solution is poured into the container, and the concentrated enzymatic hydrolysate is divided into a plurality of components with different molecular weights due to the continuous molecular weight cutoff difference among the dialysis bags.
Preferably, the crude oligosaccharide is prepared by using ice algae polysaccharide as a raw material and adopting a hydrogen peroxide method, and the specific working conditions are as follows: the Antarctic ice algae polysaccharide accounts for 0.1-3 wt% of the reaction system, the hydrogen peroxide is added in an amount of 0.1-5.0 wt% of the reaction system, the balance is water, the reaction time is 30-180 min, centrifugation is carried out after the reaction is finished, and clear liquid is collected to obtain crude oligose liquid.
The hydrogen peroxide cracking method is used for preparing the ice algae oligosaccharide, so that more obvious molecular weight concentration of the allergic substance can be obtained.
Preferably, the specific working conditions of the ultrafiltration are as follows: and (3) introducing the crude oligosaccharide solution obtained after the preparation of the crude oligosaccharide into an ultrafiltration device, wherein the membrane aperture is 5-1000 nm.
Preferably, a high-flux ultrafiltration device is used, the flux is preferably 50-100L/h, and the operating pressure difference is 50-500 kPa.
Preferably, the flash distillation has the following specific working conditions: and carrying out flash evaporation on the oligosaccharide solution obtained by ultrafiltration for 10-100 s at the temperature of 100-200 ℃.
The ultrafiltration device can be connected with the ultrahigh-temperature flash evaporation device in series, so that the working efficiency is improved. By adopting a one-step concentration method of ultrafiltration flash evaporation series connection, the effective deodorization of the ice algae oligosaccharide liquid can be realized while the concentration efficiency is improved.
The invention has the following beneficial effects:
the invention effectively removes the sensitizing substances in the Antarctic ice algae oligosaccharide by a multilayer directional dialysis technology, and simultaneously adopts a one-step concentration method of ultrafiltration flash evaporation series connection to obtain the Antarctic ice algae oligosaccharide which has the characteristics of low sensitization, no fishy smell and the like. The invention has high efficiency of the manufacturing process, easy industrialization, high oligosaccharide activity and obvious effect, simultaneously desensitizes and removes fishy smell, expands the application range of the ice algae oligosaccharide and can be widely applied to the fields of nourishment, skin care products, medicines and the like.
Drawings
FIG. 1 is a schematic view of a multi-layered directional dialysis apparatus according to the present invention;
in the figure: 1. a dialysis container; 2. a dialysis bag.
Detailed Description
The principles and features of this invention are described below in conjunction with examples, which are set forth to illustrate, but are not to be construed to limit the scope of the invention.
The Antarctic ice algae polysaccharide in the embodiment is prepared from sea antler or purple crystal algae collected from Antarctic through an alkali extraction method.
Example 1
A multilayer directional dialysis device is shown in figure 1 and comprises a dialysis container 1 with an upward opening and four dialysis bags 2 which are arranged in the container and sleeved from inside to outside in sequence according to the molecular weight cut-off, wherein the diameters of the sections of the dialysis bags 2 are increased from inside to outside in sequence.
A preparation method of low-sensitization fishy smell-removed Antarctic ice algae oligosaccharide comprises the following steps:
(1) oligosaccharide preparation: the preparation method adopts a hydrogen peroxide method to prepare the hypsizygus marmoreus oligosaccharide, and the specific working conditions are as follows: dissolving polysaccharide of the seaweed in water, and adding hydrogen peroxide for cracking; the method comprises the following steps of 1 wt% of the reaction system by using the polysaccharide of the hypsizygus marmoreus, 2 wt% of the reaction system by using the hydrogen peroxide, and the balance of water, wherein the reaction time is 100 min, centrifuging after the reaction is finished, and collecting clear liquid to obtain crude oligose liquid.
(2) Ultrafiltration and flash evaporation are connected in series for one-step concentration: connecting an ultrafiltration device and an ultrahigh-temperature flash evaporation device in series, introducing the crude oligosaccharide solution collected in the step (1) into a large-flux ultrafiltration device of 100L/h, wherein the membrane aperture is 20nm, and the operating pressure difference is 150 kPa; and (4) carrying out flash evaporation on the oligosaccharide solution after ultrafiltration at the flash evaporation temperature of 140 ℃ for 70s, and collecting flash evaporation concentrated solution.
(3) Membrane classification: the molecular weight cut-off from inside to outside of a dialysis bag in the multilayer directional dialysis device is respectively 800Da, 550Da, 350Da and 150 Da; introducing the concentrated solution obtained in the step (2) into an innermost dialysis bag of the multilayer directional dialysis device, wherein a proper amount of ultrapure water can be added into a dialysis container; the dialysis temperature is 8 ℃, and the dialysis time is 24 hours; after dialysis is finished, components outside the dialysis bag at the outermost layer and in the container are discarded, and the molecular weight ranges of the remaining four components are respectively more than 800Da, 550-800 Da, 350-550 Da and 150-350 Da.
The allergen content of each component except the component with the lowest molecular weight was measured by MTT method, and the absorbance value A (570nm) at 570nm of each component was 0.6, 1.0, 1.51, 0.8 in order from the large to the small molecular weight. Discarding the component with the largest light absorption value, namely the component of 350-550 Da, and combining the other three components to obtain the hypoallergenic fishy-removed ice algae oligosaccharide.
It should be noted that if the component with the largest absorbance value is close to the absorbance value of the adjacent component, the arrangement of the molecular weight cut-off of each dialysis bag is not reasonable and needs to be reset.
Example 2
A preparation method of low-sensitization fishy smell-removed Antarctic ice algae oligosaccharide comprises the following steps:
(1) and (3) oligosaccharide extraction: the preparation method adopts a hydrogen peroxide method to prepare the hypsizygus marmoreus oligosaccharide, and the specific working conditions are as follows: dissolving Antarctic Hairin polysaccharide in water, and adding hydrogen peroxide for cracking; the content of the polysaccharide in the reaction system is 0.1 wt%, the content of the hydrogen peroxide in the reaction system is 0.1 wt%, and the balance is water, the reaction time is 180min, after the reaction is finished, the mixture is centrifuged, and clear liquid is collected to obtain crude oligose liquid.
(2) Ultrafiltration and flash evaporation are connected in series for one-step concentration: connecting an ultrafiltration device and an ultrahigh-temperature flash evaporation device in series, introducing the crude oligosaccharide solution collected in the step (1) into a large-flux ultrafiltration device of 100L/h, wherein the membrane aperture is 1000nm, and the operating pressure difference is 50 kPa; and (3) carrying out flash evaporation on the oligosaccharide solution after ultrafiltration at the flash evaporation temperature of 100 ℃ for 100s, and collecting flash evaporation concentrated solution.
(3) Membrane classification: the multilayer directional dialysis device of example 1 was used; the molecular weight cut-off of the dialysis bag in the multilayer directional dialysis device from inside to outside is 1600Da, 1200Da, 800Da and 500Da respectively; introducing the concentrated solution obtained in the step (2) into an innermost dialysis bag of the multilayer directional dialysis device, wherein a proper amount of ultrapure water can be added into a dialysis container; the dialysis temperature is 4 ℃, and the dialysis time is 48 h; after dialysis is finished, components outside the dialysis bag at the outermost layer and in the container are discarded, and the molecular weight ranges of the remaining four components are more than 1600Da, 1200-1600 Da, 800-1200 Da and 500-800 Da respectively.
The allergen content of each component except the component with the lowest molecular weight was measured by MTT method, and the absorbance value A (570nm) at 570nm of each component was 0.84, 0.9, 1.62, 1.11 in order of molecular weight from large to small. Discarding the component with the largest light absorption value, namely the component of 800-1200 Da, and combining the other three components to obtain the low-sensitization deodorization ice algae oligosaccharide.
It should be noted that if the component with the largest absorbance value is close to the absorbance value of the adjacent component, the arrangement of the molecular weight cut-off of each dialysis bag is not reasonable and needs to be reset.
Example 3
A preparation method of low-sensitization fishy smell-removed Antarctic ice algae oligosaccharide comprises the following steps:
(1) and (3) oligosaccharide extraction: the preparation method adopts a hydrogen peroxide method to prepare the hypsizygus marmoreus oligosaccharide, and the specific working conditions are as follows: dissolving Antarctic Hairin polysaccharide in water, and adding hydrogen peroxide for cracking; 3 wt% of the polysaccharide of the hypsizygus marmoreus, 5 wt% of the hydrogen peroxide and the balance of water, wherein the reaction time is 30 min, and after the reaction is finished, centrifuging and collecting clear liquid to obtain the crude oligose liquid.
(2) Ultrafiltration and flash evaporation are connected in series for one-step concentration: connecting an ultrafiltration device and an ultrahigh-temperature flash evaporation device in series, introducing the crude oligosaccharide solution collected in the step (1) into a large-flux ultrafiltration device of 100L/h, wherein the membrane aperture is 5nm, and the operating pressure difference is 500 kPa; and (3) carrying out flash evaporation on the oligosaccharide solution after ultrafiltration at the flash evaporation temperature of 200 ℃ for 10s, and collecting flash evaporation concentrated solution.
(3) Membrane classification: the multilayer directional dialysis device of example 1 was used; the molecular weight cut-off of the dialysis bag in the multilayer directional dialysis device from inside to outside is 700Da, 500Da, 300Da and 100Da respectively; introducing the concentrated solution obtained in the step (2) into an innermost dialysis bag of the multilayer directional dialysis device, wherein a proper amount of ultrapure water can be added into a dialysis container; the dialysis temperature is 15 ℃, and the dialysis time is 12 h; after dialysis is finished, components outside the dialysis bag at the outermost layer and in the container are discarded, and the molecular weight ranges of the remaining four components are respectively more than 700Da, 500-700 Da, 300-500 Da and 100-300 Da.
The allergen content of each component except the component with the lowest molecular weight was measured by MTT method, and the absorbance value A (570nm) at 570nm of each component was 0.75, 1.75, 1.0, 0.9 in order of molecular weight from large to small. Discarding the component with the largest light absorption value, namely the component of 500-700 Da, and combining the other three components to obtain the low-allergenicity de-fishy iced algae oligosaccharide.
It should be noted that if the component with the largest absorbance value is close to the absorbance value of the adjacent component, the arrangement of the molecular weight cut-off of each dialysis bag is not reasonable and needs to be reset.
Example 4
A preparation method of low-sensitization fishy smell-removed Antarctic ice algae oligosaccharide comprises the following steps:
(1) oligosaccharide preparation: the specific working conditions for preparing the porphyridium oligosaccharide by adopting the hydrogen peroxide method are as follows: dissolving the porphyridium polysaccharide in water, and adding hydrogen peroxide for cracking; the content of the porphyridium polysaccharide in the reaction system is 2 wt%, the content of the hydrogen peroxide in the reaction system is 1.5 wt%, the balance is water, the reaction time is 120 min, centrifugation is carried out after the reaction is finished, and clear liquid is collected to obtain crude oligose liquid.
(2) Ultrafiltration and flash evaporation are connected in series for one-step concentration: connecting an ultrafiltration device and an ultrahigh-temperature flash evaporation device in series, introducing the crude oligosaccharide solution collected in the step (1) into a large-flux ultrafiltration device of 100L/h, wherein the membrane aperture is 100nm, and the operating pressure difference is 200 kPa; and (3) carrying out flash evaporation on the oligosaccharide solution after ultrafiltration at the flash evaporation temperature of 150 ℃ for 50s, and collecting flash evaporation concentrated solution.
(3) Membrane classification: the multilayer directional dialysis device of example 1 was used; the molecular weight cut-off of the dialysis bag in the multilayer directional dialysis device from inside to outside is 1200Da, 900Da, 600Da and 300Da respectively; introducing the concentrated solution obtained in the step (2) into an innermost dialysis bag of the multilayer directional dialysis device, wherein a proper amount of ultrapure water can be added into a dialysis container; the dialysis temperature is 8 ℃, and the dialysis time is 24 hours; after dialysis is finished, components outside the dialysis bag at the outermost layer and in the container are discarded, and the molecular weight ranges of the remaining four components are respectively more than 1200Da, 900-1200 Da, 600-900 Da and 300-600 Da.
The allergen content of each component except the component with the lowest molecular weight was measured by MTT method, and the absorbance value A (570nm) of each component at 570nm was 1.01, 1.14, 1.71, 0.97 in order from the large to the small molecular weight. Discarding the component with the largest light absorption value, namely the component of 600-900 Da, and combining the other three components to obtain the low-sensitization deodorization ice algae oligosaccharide.
It should be noted that if the component with the largest absorbance value is close to the absorbance value of the adjacent component, the arrangement of the molecular weight cut-off of each dialysis bag is not reasonable and needs to be reset.
Example 5
A preparation method of low-sensitization fishy smell-removed Antarctic ice algae oligosaccharide comprises the following steps:
(1) oligosaccharide preparation: the specific working conditions for preparing the porphyridium oligosaccharide by adopting the hydrogen peroxide method are as follows: dissolving the porphyridium polysaccharide in water, and adding hydrogen peroxide for cracking; the content of the porphyridium polysaccharide in the reaction system is 1 wt%, the content of the hydrogen peroxide in the reaction system is 1.5 wt%, the balance is water, the reaction time is 160 min, centrifugation is carried out after the reaction is finished, and clear liquid is collected to obtain crude oligose liquid.
(2) Ultrafiltration and flash evaporation are connected in series for one-step concentration: connecting an ultrafiltration device and an ultrahigh-temperature flash evaporation device in series, introducing the crude oligosaccharide solution collected in the step (1) into a large-flux ultrafiltration device of 100L/h, wherein the membrane aperture is 10nm, and the operating pressure difference is 150 kPa; and (3) carrying out flash evaporation on the oligosaccharide solution after ultrafiltration at the flash evaporation temperature of 150 ℃ for 50s, and collecting flash evaporation concentrated solution.
(3) Membrane classification: the multilayer directional dialysis device of example 1 was used; the molecular weight cut-off from inside to outside of a dialysis bag in the multilayer directional dialysis device is respectively 800Da, 550Da, 350Da and 150 Da; introducing the concentrated solution obtained in the step (2) into an innermost dialysis bag of the multilayer directional dialysis device, wherein a proper amount of ultrapure water can be added into a dialysis container; the dialysis temperature is 10 ℃, and the dialysis time is 20 h; after dialysis is finished, components outside the dialysis bag at the outermost layer and in the container are discarded, and the molecular weight ranges of the remaining four components are respectively more than 800Da, 550-800 Da, 350-550 Da and 150-350 Da.
The allergen content of each component except the component with the lowest molecular weight was measured by MTT method, and the absorbance value A (570nm) of each component at 570nm was 1.06, 1.88, 0.97, 0.76 in order of molecular weight from large to small. Discarding the component with the largest light absorption value, namely the component with 550-800 Da, and combining the other three components to obtain the low-allergenicity deodorization ice algae oligosaccharide.
It should be noted that if the component with the largest absorbance value is close to the absorbance value of the adjacent component, the arrangement of the molecular weight cut-off of each dialysis bag is not reasonable and needs to be reset.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A preparation method of low-sensitization deodorization Antarctic ice algae oligosaccharide is characterized by sequentially comprising crude oligosaccharide preparation, ultrafiltration, flash evaporation and membrane classification according to the preparation sequence;
the membrane fractionation is to separate the concentrated oligose solution obtained by flash evaporation into a plurality of components with different molecular weights by using a semi-permeable membrane device, detect the allergen content of each component except the component with the minimum molecular weight, discard the component with the highest allergen content and the component with the minimum molecular weight, and collect other components.
2. The method according to claim 1, wherein the concentrated oligosaccharide solution is separated into 3 to 5 fractions having different molecular weights by using a semipermeable membrane apparatus.
3. The process according to claim 2, wherein the concentrated oligosaccharide solution is separated into 5 fractions having different molecular weights by using 4 semipermeable membrane units, and the cut-off molecular weights of the 4 semipermeable membrane units are 700 to 1600Da, 500 to 1200Da, 300 to 800Da, and 100 to 500Da, respectively.
4. The method of claim 1, wherein the semipermeable membrane device is a dialysis bag.
5. The method of claim 4, wherein the semipermeable membrane comprises a plurality of dialysis bags with different cut-off molecular weights, and the dialysis bags are sequentially sleeved from inside to outside.
6. The preparation method of claim 5, wherein the dialysis bags are 4, and the cut-off molecular weights of the dialysis bags are 700-1600 Da, 500-1200 Da, 300-800 Da and 100-500 Da from inside to outside in sequence; the dialysis temperature is 4-15 ℃, and the dialysis time is 12-48 h.
7. The preparation method of claim 1, wherein the crude oligosaccharide is prepared by using Antarctic ice algae polysaccharide as raw material and adopting hydrogen peroxide method to prepare ice algae oligosaccharide;
the specific working conditions are as follows: the Antarctic ice algae polysaccharide accounts for 0.1-3 wt% of the reaction system, the hydrogen peroxide is added in an amount of 0.1-5.0 wt% of the reaction system, the balance is water, the reaction time is 30-180 min, centrifugation is carried out after the reaction is finished, and clear liquid is collected to obtain crude oligose liquid.
8. The preparation method according to claim 1, wherein the ultrafiltration is carried out under the following specific working conditions: and (3) introducing the crude oligosaccharide solution obtained after the preparation of the crude oligosaccharide into an ultrafiltration device, wherein the membrane aperture is 5-1000 nm.
9. The preparation method according to claim 8, wherein the flux of the ultrafiltration device is 50-100L/h, and the operating pressure difference is 50-500 kPa.
10. The preparation method according to claim 1, wherein the flash evaporation is carried out under the following specific working conditions: and carrying out flash evaporation on the oligosaccharide solution obtained by ultrafiltration for 10-100 s at the temperature of 100-200 ℃.
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