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

A technology for epigallocatechin and gallate, which is applied in the field of medium-pressure polymer reversed-phase chromatography to separate and purify epigallocatechin gallate, and can solve the separation performance of adsorption resin and glucan gel chromatography Poor, C18 bonded silica gel is expensive, dextran gel is expensive, etc., to achieve the effect of not easy non-characteristic adsorption, convenient for industrial continuous production, and non-polar surface

Active Publication Date: 2013-03-13
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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A 65% ethanol aqueous solution (pH=3.0, acidified with citric acid) with a mass fraction of 65% was used as the mobile phase and a tea polyphenol solution with a mass concentration of 25% was prepared. The solution was passed through a 0.45 μm microfiltration membrane, and the filtrate was collected. The filtrate was ultrasonically degassed (ultrasonic frequency 50HZ) for 30 minutes, and then allowed to stand for 10 minutes. The solution was applied to a CCTRI-1 monodisperse polymer reversed-phase chromatography column, with a sample volume of 3mL, a column length of 26cm, a column diameter of 1.2cm, a filler particle size of 10μm, a column pressure of 10Mpa, and a mobile phase for elution. At 5mL / min, the eluted fractions are collected in sections, the fractions with EGCG figure 2 As shown, the purity of the obtained EGCG is 98.69% by HPLC detection.

Embodiment 2

[0041] A 15% mass fraction of ethanol aqueous solution (pH=4.0, acidified with citric acid) was used as the mobile phase and a 25% mass concentration of tea polyphenol solution was prepared. The solution was passed through a 0.45 μm microfiltration membrane, and the filtrate was collected. The filtrate was ultrasonically degassed (ultrasonic frequency 50HZ) for 30 minutes, and then allowed to stand for 10 minutes. The solution was applied to a CCTRI-1 monodisperse polymer reversed-phase chromatography column, the sample volume was 100mL, the column length was 49cm, the column diameter was 5cm, the filler particle size was 40μm, the column pressure was 5Mpa, and the mobile phase was used for elution. The mobile phase flow rate was 100mL / min, the eluted fractions are collected in sections, the fractions with EGCG image 3 As shown, the purity of the obtained EGCG was 98.1% by HPLC detection.

Embodiment 3

[0043] A 40% mass fraction of ethanol aqueous solution (pH=5.0, acidified with citric acid) was used as the mobile phase and a 25% mass concentration of tea polyphenol solution was prepared. The solution was passed through a 0.45 μm microfiltration membrane, and the filtrate was collected. The filtrate was ultrasonically degassed (ultrasonic frequency 50HZ) for 30 minutes, and then allowed to stand for 10 minutes. The solution was applied to a CCTRI-1 monodisperse polymer reversed-phase chromatography column, the sample volume was 100mL, the column length was 48cm, the column diameter was 6.8cm, the filler particle size was 40μm, the column pressure was 2Mpa, and the mobile phase was used for elution. The mobile phase flow rate At 900mL / min, the eluted fractions are collected in sections, the fractions with EGCG Figure 4 As shown, the purity of the obtained EGCG is 97.6% by HPLC detection.

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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 catechins, in particular to a method for separating and purifying epigallocatechin gallate by medium-pressure polymer reverse phase chromatography. Background technique [0002] Epigallocatechin gallate (EGCG) is the most effective active ingredient in tea polyphenols. It belongs to catechins. It is the component with the highest content in catechins. It has strong antioxidant properties and is resistant to medicine and health care. Tumor, anti-aging, lowering blood lipids, preventing atherosclerosis, enhancing immunity and other functions; in the food industry, it can be used as antioxidant, antibacterial, fresh-keeping, and deodorant; in daily chemical products, it can be used as a special function of preservation agent, Skin care agent. Therefore, it has a wide range of uses and application prospects in medicine, food, and daily chemical industries. [0003] At present, the separation and pur...

Claims

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Application Information

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Patent Type & Authority Applications(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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