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Method for separating and purifying EGCG (Epigallocatechin Gallate) by medium-pressure high polymer inversed phase chromatography

A technology of epigallocatechin and gallate, which is applied in the field of separation and purification of epigallocatechin gallate by medium-pressure high polymer reverse-phase chromatography, and can solve the separation performance of adsorption resin and dextran gel chromatography Poor, C18 bonded silica gel is expensive, dextran gel is expensive, etc., to achieve the effect of not being easy to non-specific adsorption, convenient for industrial continuous production, and non-polar on the surface

Active Publication Date: 2014-11-05
HANGZHOU TEA RES INST CHINA COOP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Adsorption resin and dextran gel chromatography have poor separation performance and need to be combined with separation methods such as HPLC; dextran gel is expensive; high-speed countercurrent chromatography and high-performance liquid phase preparative chromatography have small preparation volumes, low separation efficiency, and require energy high consumption
Huang Yongdong and others used the B-type three-zone simulated moving bed technology, with C18 bonded silica gel as the stationary phase, ethanol and water (20:80, v / v) as the mobile phase, and the Varicol process can obtain ECG with a purity of 91.33%. The rate is 91.41% [Huang Yongdong, Jiang Heyuan, Jiang Yongwen, etc. Comparative study on the separation and purification of ECG and EGCG by traditional SMB, Varicol and Partial-discard processes [J]. Tea Science, 2011, 31(3): 201-210], but The above two methods have the disadvantages of expensive C18 bonded silica gel, a large amount of organic solvent, and a small loading capacity of the separation platform (countercurrent chromatography, moving bed).
[0005] These deficiencies restrict its application in the separation of catechins

Method used

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  • Method for separating and purifying EGCG (Epigallocatechin Gallate) by medium-pressure high polymer inversed phase chromatography
  • Method for separating and purifying EGCG (Epigallocatechin Gallate) by medium-pressure high polymer inversed phase chromatography
  • Method for separating and purifying EGCG (Epigallocatechin Gallate) by medium-pressure high polymer inversed phase chromatography

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Experimental program
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Effect test

Embodiment 1

[0039]A 65% ethanol aqueous solution (pH=3.0, acidified with citric acid) was used as the mobile phase and a 25% tea polyphenol solution was prepared. Pass the solution through a 0.45 μm microfiltration membrane and collect the filtrate. The filtrate was ultrasonically degassed (ultrasonic frequency 50HZ) for 30 minutes, and then left to stand for 10 minutes. Monodisperse polymer reversed-phase chromatographic column on solution, sample volume 3mL, column length 26cm, column diameter 1.2cm, filler particle size 10μm, column pressure 10Mpa, elution with mobile phase, mobile phase flow rate 5mL / min, the eluted fractions were collected in sections, the fractions with EGCG figure 2 As shown, detected by HPLC method, the obtained EGCG purity is 98.69%.

Embodiment 2

[0041] A 15% ethanol aqueous solution (pH=4.0, acidified with citric acid) was used as the mobile phase and a 25% tea polyphenol solution was prepared. Pass the solution through a 0.45 μm microfiltration membrane and collect the filtrate. The filtrate was ultrasonically degassed (ultrasonic frequency 50HZ) for 30 minutes, and then left to stand for 10 minutes. Monodisperse polymer reversed-phase chromatographic column on solution, sample volume 100mL, column length 49cm, column diameter 5cm, filler particle size 40μm, column pressure 5Mpa, elution with mobile phase, mobile phase flow rate 100mL / min , the fraction after elution is collected in sections, the fraction of EGCG image 3 As shown, through HPLC detection, the EGCG purity obtained is 98.1%.

Embodiment 3

[0043] A 40% ethanol aqueous solution (pH=5.0, acidified with citric acid) was used as the mobile phase and a 25% tea polyphenol solution was prepared. Pass the solution through a 0.45 μm microfiltration membrane and collect the filtrate. The filtrate was ultrasonically degassed (ultrasonic frequency 50HZ) for 30 minutes, and then left to stand for 10 minutes. Monodisperse polymer reversed-phase chromatographic column on solution, sample volume 100mL, column length 48cm, column diameter 6.8cm, filler particle size 40μm, column pressure 2Mpa, use mobile phase for elution, flow rate of mobile phase is 900mL / min, the eluted fractions were collected in sections, the fractions with EGCG Figure 4 As shown, through HPLC detection, the EGCG purity obtained is 97.6%.

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Abstract

The invention discloses a method for separating and purifying EGCG (Epigallocatechin Gallate) by medium-pressure high polymer inversed phase chromatography. The method comprises the steps of dissolving a tea polyphenol raw material by taking acidified ethanol water with certain concentration as a mobile phase, filtering the acidified ethanol water by a micro-filtration membrane, then eluting the mobile phase by using a CCTRI-1 monodisperse high polymer inversed phase chromatographic column, collecting eluted cut fractions in a segmenting manner, concentrating the cut fractions to a certain concentration by a nanofiltration membrane, and drying the concentrate in a vacuum freeze drier to obtain a high-purity EGCG product. According to the method, the process is simple, the operation is convenient, the production cycle is short, the separating efficiency is high, the product purity is high, and a solvent used in the separation and purification process is ethanol solution which is non-toxic and can be recycled, so that the low carbon and environmental protection effect is realized, and the safety problem caused by the using of organic solvents can be effectively avoided.

Description

technical field [0001] The invention relates to a method for separating and purifying catechin, in particular to a method for separating and purifying epigallocatechin gallate by medium-pressure high polymer reverse phase chromatography. Background technique [0002] Epigallocatechin gallate (EGCG) is the most effective active ingredient in tea polyphenols. It belongs to catechins and is the component with the highest content in catechins. Tumor, anti-aging, lowering blood fat, preventing atherosclerosis, enhancing immunity and other functions; in the food industry, it can be used as anti-oxidation, antibacterial, fresh-keeping, deodorant; in daily chemical products, it can be used as a quality-preserving agent with special functions, skin care agent. Therefore, it has wide application and application prospect in medicine, food and daily chemical industry. [0003] At present, the separation and purification methods of catechin monomer mainly include adsorption resin chrom...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D311/62
Inventor 李大伟朱跃进张士康张海华俞璐婷黄赟赟
Owner HANGZHOU TEA RES INST CHINA COOP
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