CN114163486A - Method for preparing oligosaccharide from pseudo-ginseng - Google Patents
Method for preparing oligosaccharide from pseudo-ginseng Download PDFInfo
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Abstract
The invention discloses a method for preparing oligosaccharide from notoginseng, which comprises (1) drying notoginseng, crushing or slicing, extracting with ethanol, filtering and concentrating to obtain extract; (2) diluting the extract with water, adsorbing and eluting with nonpolar macroporous resin, collecting column effluent and eluate, mixing, and concentrating to obtain concentrated water solution; (3) adding ethanol into the concentrated aqueous solution, refrigerating and standing, and passing through an organic ultrafiltration membrane after precipitation occurs to obtain organic ultrafiltrate; (4) passing the organic ultrafiltrate through ion exchange column, collecting effluent, and recovering ethanol to obtain oligosaccharide water solution; (5) passing the oligosaccharide aqueous solution through cation exchange resin, and collecting effluent liquid; (6) decolorizing the effluent, and freeze drying to obtain oligosaccharide. Compared with the prior art, the invention has the advantages of low cost, simple operation and higher yield and purity, and the prepared oligosaccharide does not contain galacturonic acid, methyl polygalacturonate, tannin, tanning and other substances, and is more suitable for patients with hyperacidity to take.
Description
Technical Field
The invention relates to the technical field of extraction of plant active ingredients, in particular to a method for preparing oligosaccharide from pseudo-ginseng.
Background
Oligosaccharides are also called oligosaccharides, and are a general term for low-degree polymeric saccharides having a straight chain or branched chain formed by dehydration condensation of 2 to 10 monosaccharides and glycosidic linkages, and also for saccharides having a monosaccharide number of 2 to 19, which are also called oligosaccharides, and can be classified into functional oligosaccharides and ordinary oligosaccharides. Functional oligosaccharides are widely distributed and diversified in the nature, and have the characteristics of low calorie, intestinal tract benefiting, constipation prevention, serum cholesterol reduction, organism immunity enhancement, tumor resistance and the like, so that the functional oligosaccharides become important basic raw materials and food additives in functional food production and are used by industrial production in many countries in the world.
The Notoginseng radix neutral oligosaccharide is a linear or branched chain saccharide substance formed by linking about 2-15 monosaccharides via glycoside bond. The oligosaccharide has molecular weight of 4000 or less, has immunoregulation effect, and can be used as auxiliary factor for immunostimulation, and when oligosaccharide stimulates intestinal immune cells, it can improve intestinal immunoglobulin, thereby increasing humoral and cellular immunity, and improving immune response. The applicant finds that the stomach pain of patients can be aggravated when the mixed product of the pseudo-ginseng oligosaccharide and the pseudo-ginseng amino acid is orally administrated to stomach-loss mice or old stomach disease patients take 10ml of the mixed product of the pseudo-ginseng oligosaccharide and the pseudo-ginseng amino acid orally in experiments, and according to analysis, the pseudo-ginseng oligosaccharide liquid of the crude product contains substances such as galacturonic acid, methyl polygalacturonate, tannin, tanning and the like, and the substances are acidic, sour and heavy, press sweet taste, have the effect of improving intestinal metabolism for normal people, but can aggravate the stomach pain when being taken by patients with excessive gastric acid.
The extraction and preparation methods of pseudo-ginseng oligosaccharide are reported in few documents, and the common extraction method of plant oligosaccharide is to use hot water, acid water, alkaline water and the like as solvents, reflux extraction, ultrasonic extraction and microwave extraction are used for crude extraction, the obtained crude extract is subjected to ethanol fractional precipitation of polysaccharide, so that crude product is obtained after the polysaccharide and oligosaccharide are separated, the crude product is subjected to organic solvent extraction to remove small-polarity organic matters, such as n-butanol extraction to remove saponin, and then Sevag method, organic reagent method, complexation method, microfiltration membrane interception method and the like are used for removing impurities to prepare the high-purity plant oligosaccharide. However, after the conventional water extraction method, panax notoginseng oligosaccharides are more or less coated and precipitated in the ethanol fractional precipitation process, and the higher the ethanol precipitation concentration is, the higher the oligosaccharide concentration is, the greater the oligosaccharide loss is in the process of precipitating polysaccharides, so that the yield is lower.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for preparing oligosaccharide from pseudo-ginseng, which has the advantages of low cost, simple operation, high yield and purity, and the prepared oligosaccharide does not contain galacturonic acid, methyl polygalacturonate, tannin, tanning and other substances, and is more suitable for patients with excessive gastric acid.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing oligosaccharide from Notoginseng radix comprises the following steps:
(1) drying Notoginseng radix, pulverizing or slicing, extracting with ethanol under reflux, filtering to obtain extractive solution, concentrating, and recovering ethanol to obtain extract;
(2) diluting the extract obtained in the step (1) with pure water, then passing through nonpolar macroporous resin for adsorption and water elution, collecting effluent liquid on a column, combining the effluent liquid with the eluate, and concentrating to obtain concentrated water liquid;
(3) adding ethanol into the concentrated aqueous solution obtained in the step (2) to obtain an alcohol solution, refrigerating and standing, and passing through an organic ultrafiltration membrane after precipitation occurs to obtain an organic ultrafiltrate;
(4) passing the organic ultrafiltrate obtained in the step (3) through a strong acid and strong base anion and cation mixed bed ion exchange column to exchange impurities on the anion and cation column, collecting effluent liquid, concentrating and recovering ethanol to obtain oligosaccharide aqueous solution;
(5) removing monosaccharide from the oligosaccharide aqueous solution obtained in the step (4) through strong acid cation exchange resin, and collecting effluent liquid;
(6) and (3) decoloring the effluent liquid obtained in the step (5) by using strong base type 2 anion exchange resin 202, collecting decolored liquid, and freeze-drying to obtain the oligosaccharide.
Preferably, the volume percentage of ethanol in the ethanol reflux extraction in step (1) is 60-65%.
Preferably, the nonpolar macroporous resin in the step (2) has at least one of model numbers D101, Xad-8 and Hpd 100.
Preferably, the mass ratio of the resin to the pseudo-ginseng in the step (2) is 1: 1-0.5.
Preferably, the volume fraction of ethanol in the alcoholic solution in the step (3) is 60-70%.
Preferably, the molecular weight of the organic ultrafiltration membrane in step (3) is 3000-4000.
Preferably, the types of the strong acid strong base anion-cation mixed bed ion exchange column in the step (4) are Lxp-100 and Lxp-500; the mass ratio of the cationic resin to the anionic resin to the pseudo-ginseng is 1: 1: 2.
preferably, the model of the strong acid cation exchange resin in step (5) is ZG SPC 106Ca or ZG SPC 106 Na.
The invention also provides the oligosaccharide prepared by the method.
Preferably, the monosaccharide component after oligosaccharide hydrolysis comprises 45.0-55.0% of glucose, 20.0-25.0% of fructose and 10.0-15.0% of galactose by mass percent.
The invention also provides the application of the oligosaccharide in preparing functional food.
The invention has the beneficial effects that:
(1) the invention has low cost and simple operation, the extraction rate of oligosaccharide calculated according to the mass ratio of crude drugs is more than 1 percent, the oligosaccharide has higher purity, does not contain amino acid and macromolecular polysaccharide, is all micromolecular substances, and is easier to be absorbed by human body.
(2) Compared with the prior art, the oligosaccharide prepared by the invention does not contain galacturonic acid, methyl polygalacturonate, tannin, tanning and other substances, and is more suitable for patients with excessive gastric acid to take.
Drawings
FIG. 1 is a liquid chromatogram of monosaccharide components after derivatization and hydrolysis of oligosaccharide (oligosaccharide of Panax notoginseng) prepared in example 1.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
The sources of the raw materials used in the present invention are not limited, and the raw materials used in the present invention are all those commonly available in the art unless otherwise specified.
Example 1
(1) Drying Notoginseng radix, coarse pulverizing into 100g, adding 7 times of 62% ethanol, extracting under reflux for 2 hr, filtering to obtain extractive solution, adding 5 times of 62% ethanol into the residue, extracting under reflux for 1 hr for the second time, mixing the extractive solutions, concentrating, and recovering ethanol to obtain extract;
(2) diluting the extract with 4 times of pure water, passing through 100g nonpolar macroporous resin Hpd-100 to adsorb notoginsenoside and flavone and other small-scale substances, wherein the mass ratio of the resin to the notoginseng is 1:1, collecting effluent liquid on the column, eluting the column with 1 column volume of water after sample loading, combining the effluent liquid on the column and the water-washed liquid, and concentrating to obtain concentrated water liquid 2 times of the notoginseng mass;
(3) adding 95% ethanol into the concentrated aqueous solution to make the alcohol content reach 65%, refrigerating and standing the obtained alcohol solution at 3 deg.C for 8 hr, filtering with 3000 molecular weight organic ultrafiltration membrane after precipitation occurs, and discarding the trapped precipitate to obtain organic ultrafiltrate;
(4) passing the organic ultrafiltrate through strong acid strong base anion-cation mixed bed ion exchange column (Lxp-100, Lxp-500) to exchange amino acid, acidic polysaccharide (galacturonic acid oligosaccharide), protein, part of pigment and other impurities on the anion-cation column, wherein the dosage of anion resin and cation resin is 100g each, collecting effluent liquid, concentrating and recovering ethanol to obtain non-sour oligosaccharide aqueous solution;
(5) passing the obtained oligosaccharide water solution through 100g ZG SPC 106Ca strongly acidic cation resin calcium chromatography packing to remove monosaccharide at flow rate of 50ml/h, collecting effluent, wherein the effluent is yellowish, monosaccharide components are adsorbed on calcium chromatography, and the effluent is detected by thin layer to be free of monosaccharide spots such as galactose, glucose, etc.;
(6) and (3) decoloring the effluent liquid obtained in the step (5) by using strong base type 2 anion exchange resin 202(Na type), collecting decolored liquid, freeze-drying to obtain 1.6g of oligosaccharide, and storing the oligosaccharide at minus 10 ℃ in a dark place.
The obtained oligosaccharides were subjected to derivatization reaction by HPLC method, and the contents thereof were measured as shown in fig. 1 and table 1, with glucose, fructose, and lactose as controls and glucosamine hydrochloride as an internal standard, and were found to have a glucose content of 51.25%, a fructose content of 21.41%, and a galactose content of 11.20% in terms of mass%.
TABLE 1
Example 2
(1) Drying Notoginseng radix, slicing into 100g, adding 7 times of 65% ethanol, extracting under reflux for 2 hr, filtering to obtain extractive solution, adding 5 times of 65% ethanol into the residue, extracting under reflux for 1 hr for the second time, mixing the extractive solutions, concentrating, and recovering ethanol to obtain extract;
(2) diluting the extract with 4 times of pure water, passing through 200g nonpolar macroporous resin Hpd-100 to adsorb small-scale substances such as notoginsenoside and flavone, wherein the mass ratio of the resin to the notoginseng is 1:0.5, collecting effluent liquid from column chromatography, eluting the column with 1 column volume of water after sample loading, combining the effluent liquid from column chromatography and the water washing liquid, and concentrating to obtain concentrated water liquid 2 times of the notoginseng mass;
(3) adding anhydrous ethanol into the concentrated aqueous solution to make the alcohol content reach 70%, refrigerating and standing the obtained alcohol solution at 0 ℃ for 6 hours, filtering the alcohol solution by using a 4000 molecular weight organic ultrafiltration membrane after precipitation occurs, and removing the trapped precipitate to obtain organic ultrafiltrate;
(4) passing the organic ultrafiltrate through strong acid strong base anion-cation mixed bed ion exchange column (Lxp-100, Lxp-500) to exchange amino acid, acidic polysaccharide (galacturonic acid oligosaccharide), protein, part of pigment and other impurities on the anion-cation column, wherein the dosage of anion resin and cation resin is 100g each, collecting effluent liquid, concentrating and recovering ethanol to obtain non-sour oligosaccharide aqueous solution;
(5) passing the obtained oligosaccharide water solution through 100g ZG SPC 106Na strongly acidic cation resin calcium chromatography packing to remove monosaccharide at flow rate of 50ml/h, collecting effluent, wherein the effluent is yellowish, monosaccharide components are adsorbed on calcium chromatography, and the effluent is detected by thin layer to be free of monosaccharide spots such as galactose, glucose, etc.;
(6) and (3) decoloring the effluent liquid obtained in the step (5) by using strong base type 2 anion exchange resin 202(NA type), collecting decolored liquid, freezing and drying to obtain 1.1g of oligosaccharide, and storing the oligosaccharide at minus 10 ℃ in a dark place.
And performing derivatization reaction on the obtained oligosaccharide by using an HPLC method, taking glucose, fructose and lactose as reference substances and glucosamine hydrochloride as an internal standard substance, and determining the content of the oligosaccharide, wherein the content of the glucose is 44.96%, the content of the fructose is 24.61% and the content of the galactose is 14.60% in percentage by mass.
Example 3
(1) Drying Notoginseng radix, coarse pulverizing into 100g, adding 7 times of 60% ethanol, extracting under reflux for 2 hr, filtering to obtain extractive solution, adding 5 times of 60% ethanol into the residue, extracting under reflux for 1 hr for the second time, mixing the extractive solutions, concentrating, and recovering ethanol to obtain extract;
(2) diluting the extract with 4 times of pure water, passing through 100g nonpolar macroporous resin Hpd-100 to adsorb notoginsenoside and flavone and other small-scale substances, wherein the mass ratio of the resin to the notoginseng is 1:0.7, collecting effluent liquid on the column, eluting the column with 1 column volume of water after sampling, combining the effluent liquid on the column and the water eluate, and concentrating to obtain concentrated water liquid 2 times of the notoginseng mass;
(3) adding 95% ethanol into the concentrated aqueous solution to make the alcohol content reach 60%, refrigerating and standing the obtained alcohol solution at 3 deg.C for 8 hr, filtering with 3000 molecular weight organic ultrafiltration membrane after precipitation occurs, and discarding the trapped precipitate to obtain organic ultrafiltrate;
(4) passing the organic ultrafiltrate through strong acid strong base anion-cation mixed bed ion exchange column (Lxp-100, Lxp-500) to exchange amino acid, acidic polysaccharide (galacturonic acid oligosaccharide), protein, part of pigment and other impurities on the anion-cation column, wherein the dosage of anion resin and cation resin is 100g each, collecting effluent liquid, concentrating and recovering ethanol to obtain non-sour oligosaccharide aqueous solution;
(5) passing the obtained oligosaccharide water solution through 100g ZG SPC 106Ca strongly acidic cation resin calcium chromatography packing to remove monosaccharide at flow rate of 50ml/h, collecting effluent, wherein the effluent is yellowish, monosaccharide components are adsorbed on calcium chromatography, and the effluent is detected by thin layer to be free of monosaccharide spots such as galactose, glucose, etc.;
(6) and (3) decoloring the effluent liquid obtained in the step (5) by using strong base type 2 anion exchange resin 202(Na type), collecting decolored liquid, freeze-drying to obtain 1.2g of oligosaccharide, and storing the oligosaccharide at minus 10 ℃ in a dark place.
Performing derivatization reaction on the obtained oligosaccharide by using an HPLC method, taking glucose, fructose and lactose as reference substances and glucosamine hydrochloride as an internal standard substance, and determining the content of the oligosaccharide, wherein the content of the oligosaccharide is 54.75 percent, the content of the fructose is 20.05 percent and the content of the galactose is 10.00 percent in percentage by mass.
Comparative example 1
This comparative example differs from example 1 in that:
(1) drying Notoginseng radix, coarse pulverizing into 100g, adding 15 times of water, reflux-extracting for 2 hr, filtering to obtain extractive solution, adding 5 times of water into the residue, reflux-extracting for another time for 1 hr, mixing the extractive solutions, and concentrating to obtain extract;
(2) diluting the extract with 4 times of pure water, passing through 100g nonpolar macroporous resin Hpd-100 to adsorb notoginsenoside and flavone and other small-scale substances, wherein the mass ratio of the resin to the notoginseng is 1:1, collecting effluent liquid on the column, eluting the column with 1 column volume of water after sample loading, combining the effluent liquid on the column and the water-washed liquid, and concentrating to obtain concentrated water liquid 2 times of the notoginseng mass;
(3) adding 95% ethanol into the concentrated water solution to make the alcohol content reach 75%, refrigerating and standing the obtained alcohol solution at 3 deg.C for 12 hr, filtering with 3000 molecular weight organic ultrafiltration membrane after precipitation occurs, and discarding the trapped precipitate to obtain organic ultrafiltrate;
(4) passing the organic ultrafiltrate through strong acid strong base anion-cation mixed bed ion exchange column (Lxp-100, Lxp-500) to exchange the impurities of amino acid, acidic polysaccharide (galacturonic acid oligosaccharide), protein, partial pigment, etc. on the anion-cation column, anion resin 100g, cation resin 200g, collecting effluent liquid to obtain non-sour oligosaccharide aqueous solution;
(5) passing the obtained oligosaccharide water solution through 100g DEAE series cellulose column to remove monosaccharide at flow rate of 50ml/h, collecting effluent liquid, wherein the effluent liquid is yellowish, monosaccharide components are adsorbed on the filler, and the effluent liquid is checked by thin layer to be free of monosaccharide spots such as galactose, glucose, etc.;
(6) and (3) decoloring the effluent liquid obtained in the step (5) by using strong base type 2 anion exchange resin 202(Na type), collecting decolored liquid, freeze-drying to obtain 0.8g of oligosaccharide, and storing the oligosaccharide at minus 10 ℃ in a dark place.
And performing derivatization reaction on the obtained oligosaccharide by using an HPLC (high performance liquid chromatography) method, taking glucose, fructose and lactose as reference substances and taking glucosamine hydrochloride as an internal standard substance, and determining the content of the oligosaccharide, wherein the content of glucose is 64.13 percent, the content of fructose is 10.61 percent and the content of galactose is 7.24 percent in percentage by mass.
This comparative example is extracted with water, 7 times water extraction 100g medicinal materials, a large amount of cladding, dense state, the composition is mentioned incompletely, and be difficult to filter, therefore, the quantity of water has been improved, it can basically be extracted to ensure macromolecular polysaccharide, when step (2), the polysaccharide flows out along macroporous resin upper sample liquid, concentrate, the concentration time is long, concentrate pH is basically about 4, the concentrate has some sour, this acidity can be confirmed to be brought by amino acid and galacturonic acid, heat the concentration in-process simultaneously, can hydrolyze some macromolecular polysaccharides, after to last end product glucose higher, fructose and galactose can reduce.
Meanwhile, the oligosaccharide prepared by the comparative example is used for replacing pseudo-ginseng oligosaccharide to prepare a product mixed by oligosaccharide and pseudo-ginseng amino acid, and adverse reactions such as stomachache and the like can be caused when a mouse with stomach loss by oral administration or a patient with old stomach diseases takes 10ml per day.
Comparative example 2
This comparative example differs from example 1 in that:
(1) drying Notoginseng radix, coarse pulverizing into 100g, adding 7 times of 55% ethanol, reflux-extracting for 2 hr, filtering to obtain extractive solution, adding 5 times of 55% ethanol into the residue, reflux-extracting for the second time for 1 hr, mixing the extractive solutions, and concentrating to obtain extract;
(2) diluting the extract with 4 times of pure water, passing through 100g nonpolar macroporous resin Hpd-100 to adsorb notoginsenoside and flavone and other small-scale substances, wherein the mass ratio of the resin to the notoginseng is 1:1, collecting effluent liquid on the column, eluting the column with 1 column volume of water after sample loading, combining the effluent liquid on the column and the water-washed liquid, and concentrating to obtain concentrated water liquid 2 times of the notoginseng mass;
(3) adding 95% ethanol into the concentrated aqueous solution to make the ethanol content reach 55%, refrigerating and standing the obtained alcohol solution at 3 deg.C for 12 hr, filtering with 3000 molecular weight organic ultrafiltration membrane after precipitation occurs, and discarding the trapped precipitate to obtain organic ultrafiltrate;
(4) passing the organic ultrafiltrate through strong acid strong base anion-cation mixed bed ion exchange column (Lxp-100, Lxp-500) to exchange the impurities of amino acid, acidic polysaccharide (galacturonic acid oligosaccharide), protein, partial pigment, etc. on the anion-cation column, anion resin 50g, cation resin dosage 100g, collecting effluent liquid to obtain slightly acidic and sweet oligosaccharide aqueous solution;
(5) passing the obtained oligosaccharide water solution through 100g ZG SPC 106Na strongly acidic cation resin calcium chromatographic packing to remove monosaccharide at flow rate of 50ml/h, collecting effluent, wherein the effluent is slightly yellowish, sour and sweet, monosaccharide components are adsorbed on the packing, and the effluent is detected by thin layer to have no monosaccharide spots such as galactose, glucose, etc.;
(6) and (3) decoloring the effluent obtained in the step (5) by using strong base type 2 anion exchange resin 202(Na type), collecting decolored liquid which is acid-free sweet light yellow liquid, freezing and drying to obtain 1.5g of oligosaccharide, and storing the oligosaccharide in the dark at minus 10 ℃.
And performing derivatization reaction on the obtained oligosaccharide by using an HPLC method, taking glucose, fructose and lactose as reference substances and glucosamine hydrochloride as an internal standard substance, and determining the content of the oligosaccharide, wherein the content of the glucose is 32.43 percent, the content of the fructose is 6.23 percent and the content of the galactose is 4.54 percent in percentage by mass.
The invention directly adopts 60-65% ethanol to extract the panax notoginseng medicinal tablets, has higher oligosaccharide yield, less panax notoginseng polysaccharide and higher oligosaccharide purity, and avoids the problem of large ethanol consumption caused by oligosaccharide coating when the traditional water extraction method is used for alcohol precipitation.
When organic matters of notoginsenoside and notoginsenones are removed, a nonpolar macroporous resin method is adopted to replace an n-butanol extraction method, the operation is more convenient, the nonpolar resin has low polarity and is not easy to adsorb sugar substances, and the damage of oligosaccharide is avoided. The obtained sugar substances are arranged in high and low content and mainly contain substances such as oligosaccharide, amino acid, starch, micromolecular polypeptide, protein and the like, ethanol is added to ensure that the alcohol content is 60-70%, the substances are refrigerated and kept stand at low temperature of 0-3 ℃, a small amount of precipitates are generated, the precipitates are mainly starch, protein, tanning and the like, and the precipitates are more easily separated out and more completely precipitated in low-temperature ethanol solution than conventional aqueous solution low-temperature or freeze-dissolving methods.
In addition, a 3000-and 4000-molecular weight organic membrane is adopted for filtering, macromolecular polysaccharide substances are removed through interception, and the organic ultrafiltrate containing the micromolecular oligosaccharide which flows out through filtration is collected. The invention finds that the alcohol solution after precipitation is easier than the concentrated water solution directly filtered, and the filtering time is shortened by 1/2.
Organic ultrafiltrate containing micromolecule oligosaccharide is passed through anion and cation to remove impurities of amino acid, acidic polysaccharide, polypeptide, protein, tanning, phenolic acid, pigment and the like, compounds with acidic groups are exchanged onto anion, substances with amino ions are exchanged onto cation, simultaneously, effluent liquid passing through anion and cation columns is subjected to ethanol recovery, on one hand, the loss of oligosaccharide in the adsorption process can be reduced, on the other hand, the oligosaccharide and monosaccharide can be separated by adopting domestic common strong acid GA chromatographic resin filler under the condition of a water system, and compared with DEAE series cellulose columns, the separation cost is extremely low, and the operation is simpler.
Meanwhile, the oligosaccharide prepared by the invention is used for replacing pseudo-ginseng oligosaccharide, and a mixed product of the oligosaccharide and pseudo-ginseng amino acid is prepared, so that the stomach-perfusion stomach-loss mouse can be orally taken, or 10ml of the oligosaccharide can be orally taken by a patient with old stomach diseases every day, and stomachache can not be generated. The result shows that the oligosaccharide prepared by the invention is more suitable for people with hyperacidity, and avoids the side effect of traditional pseudo-ginseng oligosaccharide on stomachache of people with hyperacidity.
In conclusion, the method for preparing oligosaccharide from pseudo-ginseng provided by the invention has the advantages of low cost, simple operation and higher yield and purity, and compared with the prior art, the oligosaccharide prepared by the invention does not contain galacturonic acid, polygalacturonic acid methyl ester, tannin, tanning and other substances, and is more suitable for patients with excessive gastric acid.
The present invention has been further described with reference to specific embodiments, which are only exemplary and do not limit the scope of the present invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.
Claims (10)
1. A method for preparing oligosaccharide from pseudo-ginseng is characterized by comprising the following steps:
(1) drying Notoginseng radix, pulverizing or slicing, extracting with ethanol under reflux, filtering to obtain extractive solution, concentrating, and recovering ethanol to obtain extract;
(2) diluting the extract obtained in the step (1) with pure water, then passing through nonpolar macroporous resin for adsorption and water elution, collecting effluent liquid on a column, combining the effluent liquid with the eluate, and concentrating to obtain concentrated water liquid;
(3) adding ethanol into the concentrated aqueous solution obtained in the step (2) to obtain an alcohol solution, refrigerating and standing, and passing through an organic ultrafiltration membrane after precipitation occurs to obtain an organic ultrafiltrate;
(4) passing the organic ultrafiltrate obtained in the step (3) through a strong acid and strong base anion and cation mixed bed ion exchange column to exchange impurities on the anion and cation column, collecting effluent liquid, concentrating and recovering ethanol to obtain oligosaccharide aqueous solution;
(5) removing monosaccharide from the oligosaccharide aqueous solution obtained in the step (4) through strong acid cation exchange resin, and collecting effluent liquid;
(6) and (3) decoloring the effluent liquid obtained in the step (5) by using strong base type 2 anion exchange resin 202, collecting decolored liquid, and freeze-drying to obtain the oligosaccharide.
2. The process of claim 1, wherein the ethanol reflux extraction of step (1) comprises 60-65% by volume ethanol.
3. The method of claim 1, wherein the non-polar macroporous resin of step (2) is at least one of type D101, Xad-8, and Hpd 100.
4. The method according to claim 1, wherein the volume fraction of ethanol in the alcoholic solution in the step (3) is 60-70%.
5. The method as claimed in claim 4, wherein the molecular weight of the organic ultrafiltration membrane in step (3) is 3000-4000.
6. The method as claimed in claim 1, wherein the strong acid strong base anion and cation mixed bed ion exchange column in step (4) has model numbers of Lxp-100 and Lxp-500, wherein the mass ratio of cation resin, anion resin and notoginseng is 1: 1: 2.
7. the process of claim 1, wherein the strong acid cation exchange resin type in step (5) is ZG SPC 106Ca or ZG SPC 106 Na.
8. An oligosaccharide obtainable by a process according to any one of claims 1 to 7.
9. The oligosaccharide of claim 8, wherein the hydrolyzed monosaccharide composition comprises, in mass percent, 45.0-55.0% glucose, 20.0-25.0% fructose, and 10.0-15.0% galactose.
10. Use of an oligosaccharide according to claim 9 in the preparation of a functional food.
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