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Method for separating and purifying epigallocatechin gallate by aqueous two-phase system

A technology of epigallocatechin and gallate, which is applied in the field of biochemical industry, can solve the problems of improving and detrimental to the purity of catechin, and achieve the effect of high purity, simple operation and easy handling

Inactive Publication Date: 2014-05-21
北京起航诚业生物科技中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At a lower concentration (NaCl<5%), an appropriate increase in salt concentration is beneficial to the increase of ester-soluble catechins (such as EGCG) and the total amount of catechins, but after reaching a certain salt concentration, the increase in concentration is not conducive to Improvement of catechin purity (Chinese Tea, 1999, (4): 29-29)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] 1) Dosing: select catechin with an EGCG content of 60% as a raw material, weigh 0.05 g, and dissolve it with 10 mL of 30% ethanol aqueous solution to obtain a dark tea-red catechin liquid.

[0020] 2) Separation of two aqueous phases: using acetone / K 2 HPO 4 The two-phase system extracts EGCG, and its preparation process is: add 0.25g K to each milliliter of water 2 HPO 4 , stirred to fully dissolve the phosphate, and then add acetone to form a two-phase aqueous system, the volume ratio of acetone to water is 8:2. Add catechin liquid in the two-phase system, adjust the pH value with 10% hydrochloric acid, make the pH value in the range of 6-7, separate with a centrifuge, and the two-phase partition coefficient (K=C 有机相 / C 水 相 ) is 4.18, the yield Y=(C of EGCG in the organic phase 有机相 ×V 有机相 ) / 50mg×60%=98.43%.

[0021] 3) Concentration and drying: the acetone phase was separated by a separatory funnel, and 29.33 mg of yellow solid crystals were obtained by rotary...

Embodiment 2

[0023] 1) Dosing: select catechin with an EGCG content of 60% as a raw material, weigh 0.5 g, and dissolve it with 10 mL of 30% ethanol aqueous solution to obtain a dark tea-red catechin liquid.

[0024] 2) Separation of two aqueous phases: using acetone / K 2 HPO 4 The two-phase system extracts EGCG, and its preparation process is: add 0.5g K 2 HPO 4 , Stir to fully dissolve the phosphate, then add acetone to form a two-phase aqueous system, the volume ratio of acetone to water is 7:3. Add catechin liquid in the two-phase system, adjust the pH value with 10% hydrochloric acid, make the pH value in the range of 6-7, and separate it with a centrifuge. The two-phase partition coefficient is 2.32. Rate Y=95.53%.

[0025] 3) Concentration and drying: the acetone phase was separated by a separatory funnel, and 23.83 mg of yellow solid crystals were obtained by rotary evaporation, and the purity of EGCG was 80.98%.

Embodiment 3

[0027] 1) Dosing: select catechin with an EGCG content of 60% as a raw material, weigh 0.5 g, and dissolve it with 10 mL of 30% ethanol aqueous solution to obtain a dark tea-red catechin liquid.

[0028] 2) Separation of two aqueous phases: using acetone / K 2 HPO 4 The two-phase system extracts EGCG, and its preparation process is: add 0.75g K to every milliliter of water 2 HPO 4 , Stir to fully dissolve the phosphate, then add acetone to form a two-phase aqueous system, the volume ratio of acetone to water is 6:4. Add catechin medicinal solution to the two-phase aqueous system, adjust the pH value with 10% hydrochloric acid, make the pH value in the range of 6-7, separate with a centrifuge, the two-phase aqueous phase partition coefficient is 2.59, and the collection of EGCG in the organic phase Rate Y=91.81%.

[0029]3) Concentration and drying: the acetone phase was separated by a separatory funnel, and 21.86 mg of yellow solid crystals were obtained by rotary evaporatio...

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Abstract

The invention discloses a method for separating and purifying epigallocatechin gallate by an aqueous two-phase system. The method comprises following steps of (1) preparing a liquor: dissolving the raw material catechinic acid with the content of epigallocatechin gallate being 40-70% in ethyl alcohol aqueous solution so as to obtain dark umber solution; (2) carrying out aqueous two-phase separating: dissolving dipotassium phosphate in water, then adding an organic solvent into the solution so as to form the aqueous two-phase system, adding the solution obtained in step (1) into the aqueous two-phase system, regulating the pH value to be 6-7 with 5-15% hydrochloric acid solution, and centrifugally separating an organic phase from a water phase for 2-5 minutes by a centrifugal machine at the rotating speed of 1000-2000rpm, wherein the EGCG (epigallocatechin gallate) is gathered in the organic phase, and impurities are gathered in water phase; (3) concentrating and drying: removing the solvent through rotary evaporation of the organic phase so as to obtain yellow solid crystal. The epigallocatechin gallate obtained by the method has high purity, and can easily meet the food safety requirement according to impurity detection.

Description

technical field [0001] The invention relates to a method for separating and purifying epigallocatechin gallate in a two-phase system formed by common organic solvents and brine, and belongs to the field of biochemical industry. Background technique [0002] Aqueous two-phase extraction technology means that the aqueous solution of hydrophilic polymer can form two aqueous phases under certain conditions, and the separation can be realized due to the different distribution of the separated substances in the two phases. This technology is widely used in the separation and extraction of products in the fields of biochemistry, cell biology and biochemical industry. Aqueous two-phase extraction technology requires less investment in equipment and is easy to operate. Most of these two-phase aqueous systems are polyethylene glycol / glucose and polyethylene glycol / inorganic salts. Since water-soluble polymers are difficult to volatilize, stripping is necessary, and salt enters the st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D311/62
CPCC07D311/62
Inventor 霍清杨晓芳
Owner 北京起航诚业生物科技中心