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CN-121975034-A - Method for extracting and separating polysaccharide in radix isatidis by using eutectic solvent aqueous two-phase system

CN121975034ACN 121975034 ACN121975034 ACN 121975034ACN-121975034-A

Abstract

The invention discloses a method for extracting and separating polysaccharide in radix isatidis by using a eutectic solvent aqueous two-phase system, which comprises the steps of adding radix isatidis powder into an aqueous solution of the eutectic solvent, mixing a hydrogen bond donor with a hydrogen bond acceptor and/or deionized water to obtain the eutectic solvent, centrifuging to obtain a supernatant fluid, obtaining a crude extract of radix isatidis polysaccharide, and then adding monopotassium phosphate into the mixed crude extract to form an aqueous two-phase extraction system, wherein the aqueous solution of monopotassium phosphate is polysaccharide. The method for extracting and separating the isatis root polysaccharide is simple and efficient, and has the characteristics of high extraction rate, mild conditions, good selectivity, green and economical property.

Inventors

  • LI YUXI
  • YU CHANGJIANG
  • MENG CHUNYU
  • ZHANG JIAMING
  • KONG DEJING
  • LI JUNBAI
  • Krist Homberg

Assignees

  • 云南昆平实业有限公司
  • 中国科学院青岛生物能源与过程研究所

Dates

Publication Date
20260505
Application Date
20260213

Claims (9)

  1. 1. A method for extracting and separating radix isatidis polysaccharide by using a eutectic solvent aqueous two-phase system is characterized by comprising the following steps: 1. preparing a eutectic solvent; 2. Preparing a eutectic solvent aqueous solution; 3. Forming a crude extract of the polysaccharide of the radix isatidis; 4. forming a double-water phase extraction system, and extracting and separating.
  2. 2. The method of claim 1, wherein in the first step, the eutectic solvent is formed by mixing a hydrogen bond donor and a hydrogen bond acceptor, The hydrogen bond donor is selected from one or more of lactic acid, ethylene glycol, 1, 2-propylene glycol, acetamide, urea, methyl urea, N '-dimethyl urea, glycerol, sorbitol and malic acid, preferably selected from lactic acid, ethylene glycol, 1, 2-propylene glycol, acetamide, urea, methyl urea and N, N' -dimethyl urea; the hydrogen bond acceptor is one or more of choline chloride, betaine, L-proline and D-proline, preferably choline chloride or betaine.
  3. 3. The method of claim 2, wherein, in step one, The ratio of the amounts of the hydrogen bond donor and the hydrogen bond acceptor is 1:0.1 to 10, preferably 1:0.25 to 4, more preferably 1:0.5 to 2.
  4. 4. The method of claim 3, wherein in step one, The formulation of the eutectic solvent is carried out at room temperature or above, preferably by heating, for example to 30-95 ℃, preferably 50-90 ℃, more preferably 70-85 ℃.
  5. 5. Method according to one of claims 1 to 4, characterized in that in step two, the eutectic solvent is mixed with water, preferably deionized water, and mixed homogeneously under stirring, obtaining an aqueous solution of the eutectic solvent.
  6. 6. The method according to any one of claims 1 to 5, wherein in step three, the radix isatidis powder is added into the aqueous solution of the eutectic solvent, and the crude extract of the radix isatidis polysaccharide is obtained by ultrasonic extraction at 20-60 ℃ for 20-60min, centrifugation and supernatant extraction.
  7. 7. The process according to any one of claims 1 to 6, wherein in step four, potassium dihydrogen phosphate is added to the mixed crude extract to form a two aqueous phase extraction system.
  8. 8. The method of claim 7, wherein in step four, the eutectic solvent phase is protein and other impurities and the aqueous potassium dihydrogen phosphate phase is polysaccharide.
  9. 9. The method according to claim 7 or 8, wherein in step four, Adding inorganic salt, preferably potassium dihydrogen phosphate, into the crude extract, wherein the mass-to-volume ratio (g/ml) of the inorganic salt to the crude extract is 1:2-10, preferably 1:3-5, standing after the inorganic salt is completely dissolved, and centrifuging to form a double-water-phase extraction system.

Description

Method for extracting and separating polysaccharide in radix isatidis by using eutectic solvent aqueous two-phase system Technical Field The invention belongs to the field of extraction of active ingredients of natural products, and particularly relates to a preparation method of an extract of radix isatidis, in particular to a method for extracting and separating radix isatidis polysaccharide by using a eutectic solvent aqueous two-phase system. Background The radix Isatidis is dry root and rhizome of herba Kalimeridis (Baphicacanthuscusia (Nees) Bremek) of Acanthaceae, has bitter taste and cold nature, and can restore heart and stomach channels, has effects of clearing heat and detoxicating, cooling blood and resolving macula, and can be clinically used for epidemic warm syndrome with toxic heat, pharyngalgia, epidemic warm toxin with macula and erysipelas. Is widely distributed in China, especially in Guangxi and Fujian, and in southern areas, the Chinese medicinal composition is used as a medicament. The active substance of radix Isatidis is mainly polysaccharide. The extraction of active substances is the key of the production process of the traditional Chinese medicine, and the selection of the extraction solvent lays a foundation for the achievement of the link. The extraction solvent has not only about the extraction efficiency of the active ingredients, but also the profound effect on the quality of the Chinese medicine, and has a non-negligible effect on the aspect of Chinese medicine safety. In the current production and research of traditional Chinese medicines, common extraction solvents are water and alcohol solvents (such as ethanol with different proportions). The two solvents have the advantages of easily available raw materials, small toxic and side effects and the like, but also have the problems of low extraction efficiency, weak extraction capacity for certain types of active ingredients, poor extraction selectivity and the like, and are needed to be solved. Therefore, a method for extracting and separating the polysaccharide of the radix isatidis with high efficiency, green and high extraction rate is urgently needed. Disclosure of Invention Aiming at the problems in the prior art, the invention provides a method for extracting and separating the isatis root polysaccharide by using a eutectic solvent aqueous two-phase system, and the method for preparing the traditional Chinese medicine extract by introducing the eutectic solvent has the characteristics of mild condition, high efficiency, low cost, high extraction rate and environment friendliness. In order to achieve the above object, in a first aspect, the present invention provides a method for extracting and separating radix Isatidis polysaccharide by using eutectic solvent aqueous two-phase system, comprising the following steps: 1. preparing a eutectic solvent; 2. Preparing a eutectic solvent aqueous solution; 3. Forming a crude extract of the polysaccharide of the radix isatidis; 4. forming a double-water phase extraction system, and extracting and separating. In the first step of the present invention, the eutectic solvent is formed by mixing a hydrogen bond donor and a hydrogen bond acceptor. The hydrogen bond donor may be one or more selected from lactic acid, ethylene glycol, 1, 2-propanediol, acetamide, urea, methyl urea, N ' -dimethylurea, glycerol, sorbitol, malic acid, preferably lactic acid, ethylene glycol, 1, 2-propanediol, acetamide, urea, methyl urea, N ' -dimethylurea, more preferably urea or N, N ' -dimethylurea. The hydrogen bond acceptor is one or more of choline chloride, betaine, L-proline and D-proline, preferably choline chloride or betaine. Preferably, the ratio of the amounts of the substances of the hydrogen bond donor to the hydrogen bond acceptor is 1:0.1 to 10, preferably 1:0.25 to 4, more preferably 1:0.5 to 2, for example, the ratio of the amounts of the substances of the hydrogen bond donor to the hydrogen bond acceptor is 1:1 or 1:2, and the eutectic solvent is prepared by mixing and heating. According to a preferred embodiment, the formulation of the eutectic solvent may be carried out at room temperature, preferably with heating, for example to 30-95 ℃, preferably to 50-90 ℃, more preferably to 70-85 ℃. Preferably, the eutectic solvent is prepared by mixing the hydrogen bond donor with the hydrogen bond acceptor and/or deionized water and then heating and stirring at 75-82 ℃ for several minutes to several hours, preferably 15-45 minutes, to form a clear and uniform eutectic solvent. In the second step of the invention, the eutectic solvent is mixed with water, preferably deionized water, and uniformly mixed under stirring to obtain the eutectic solvent aqueous solution. Preferably, the mixture of hydrogen bond donor and hydrogen bond acceptor is mixed with deionized water in a volume ratio of 2-8:1, preferably 3-5:1, e.g. 4:1, and prepared under heating. In the invention, the eutectic solvent is