Method for capturing and separating effective components in liquorice by utilizing mixed-mode agarose gel medium

A technology of agarose gel and mixed mode, which is applied in the field of chemical engineering to achieve the effects of reducing operation procedures, simplifying operation procedures, and high purity and yield

Active Publication Date: 2020-04-14
TAIZHOU GUOKEHUAWU BIOMEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, the use of agarose gel packing to capture and separate the effective components in licorice is still a new field, and the related methods need to be further developed and perfected

Method used

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  • Method for capturing and separating effective components in liquorice by utilizing mixed-mode agarose gel medium
  • Method for capturing and separating effective components in liquorice by utilizing mixed-mode agarose gel medium
  • Method for capturing and separating effective components in liquorice by utilizing mixed-mode agarose gel medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The sample is aqueous extract of licorice, which is spray-dried into powder after being clarified and filtered. When in use, the extract powder is configured as a 25 mg / mL aqueous solution, and it is ready to use after passing through a 0.45 μm membrane. 45 mL of TREN mixed-mode agarose gel medium from Bio-works was used to pack the column for capture and separation. The specific operation process is as follows:

[0045] Loading solution: 25mg / mL licorice extract solution

[0046] Loading volume: 400mL

[0047] Packed column: TREN (45mL)

[0048] Eluent: A: Phosphate buffer solution (pH=5), B: Phosphate buffer solution (pH=10)

[0049] Flow rate: 10mL / min

[0050] Experimental steps:

[0051] Balance the prepacked column, inject the sample after the balance is completed, rinse with purified water after the injection, elute with eluent A after rinsing, rinse with water after elution, and elute with eluent B, no base elution after the peak , until no peak ends. Th...

Embodiment 2

[0053] The sample is the 60% ethanol aqueous solution extract of licorice and coptis, which is sprayed and dried into powder after clarification and filtration. When in use, prepare the extract powder as a 30 mg / mL aqueous solution, and pass through a 0.45 μm membrane before use. 5 mL of Captocore 700 mixed-mode agarose gel medium from GE Company was used to pack the column for capture and separation.

[0054] The specific operation process is as follows:

[0055] Loading solution: 30mg / mL licorice and coptis extract solution

[0056] Loading volume: 50mL

[0057] Packed column: Capto core 700 5mL prepacked column

[0058] Eluent: A: Phosphate buffer solution (pH=5), B: Phosphate buffer solution (pH=10)

[0059] Flow rate: 5mL / min

[0060] Experimental steps:

[0061] Balance the prepacked column, inject the sample after the balance is completed, rinse with purified water after the injection, elute with eluent A after rinsing, rinse with water after elution, and elute wi...

Embodiment 3

[0063] The sample is the crude extract of liquiritin, wherein the content of liquiritin is 40%, which is spray-dried into powder after being clarified and filtered. When in use, the extract powder is configured as a 25 mg / mL ethanol solution, and it is ready to use after passing through a 0.45 μm membrane. Take 5 mL of TREN mixed-mode agarose gel medium from Bio-works for capture and separation. Place the TREN medium in the ethanol solution of the crude extract of liquiritin, mix well and place it on a shaker for 8 hours, let it stand, filter the agarose gel medium, and fill it into a 5mL empty column tube such as SPE.

[0064] Eluent: A: Phosphate buffer solution (pH=5), B: Phosphate buffer solution (pH=10)

[0065] Experimental steps:

[0066] Equilibrate the packing, rinse with purified water after the equilibrium is completed, elute with eluent A after elution, rinse with water after elution of 3 column volumes, elute with eluent B, and elute with alkali after elution of...

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Abstract

The invention relates to a method for capturing and separating effective components in liquorice by utilizing a mixed-mode agarose gel medium. The method is characterized in that the mixed-mode agarose gel medium with ion exchange and hydrophobic ligands is used for capturing and separating acidic saponin substances and flavonoid substances in liquorice; the two substances are effectively adsorbedin the agarose gel medium in an adsorption mode, and effective separation of the acidic saponin substances and the flavonoid substances is realized by changing the pH value of an eluent. According tothe method for capturing and separating liquiritin and glycyrrhizic acid substances in liquorice, the result can approximately reach that the purity of the liquiritin substances is 65%-95%, and the yield is 80%-98% (calculated by taking the liquiritin as a standard substance); the purity of glycyrrhizic acid substances is 60-95%, the yield is 75-95% (calculated by taking ammonium glycyrrhizinateas a standard substance), the purity and the yield are relatively high, and the method has important significance for a component separation process of natural products and a traditional Chinese medicine extraction and purification process.

Description

technical field [0001] The invention relates to the field of chemical engineering, in particular to a method for capturing and separating effective components in licorice using a mixed-mode agarose gel medium. Background technique [0002] Licorice (scientific name Glycyrrhiza uralensis Fisch), aliases: Guolao, sweet grass, Ural licorice, sweet root. Legumes and licorice are perennial herbs with strong roots and rhizomes. They are a kind of tonic Chinese herbal medicine. [0003] When used as a traditional Chinese medicine, licorice generally refers to the dried roots and rhizomes of Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L. of leguminous plants. The root is cylindrical, 25-100 cm long, and 0.6-3.5 cm in diameter. The outer skin is different in elasticity, and the surface is reddish brown or grayish brown. The rhizome is cylindrical, with bud scars on the surface and pith in the middle of the section. Slight smell, sweet and special....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/42B01D15/08
CPCB01D15/08B01D15/424
Inventor 梁鑫淼于伟郭志谋刘坤董佶
Owner TAIZHOU GUOKEHUAWU BIOMEDICAL TECH CO LTD
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