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Separation purification process for main catechin component in tea polyphenol and glycosidase activity

A technology for separation and purification of tea polyphenols, which is applied in the direction of organic active ingredients, medical preparations containing active ingredients, organic chemistry, etc. simple effect

Inactive Publication Date: 2009-07-29
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The above-mentioned Sephadex LH-20 method and HPLC method have high cost and long period

Method used

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  • Separation purification process for main catechin component in tea polyphenol and glycosidase activity
  • Separation purification process for main catechin component in tea polyphenol and glycosidase activity
  • Separation purification process for main catechin component in tea polyphenol and glycosidase activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Weigh 300 grams of a tea polyphenol extract with an epigallocatechin gallate content of 50%, and add 1200 mL of deionized water to dissolve it. Take 4kg of HPD100 macroporous adsorption resin and put it into the column with 95% ethanol, wash away the porogen and polymerized monomer residue in the resin with 8L of 95% ethanol, until the eluate is concentrated and qualified by gas phase detection. Then replace the ethanol in the column with deionized water, and load the above-mentioned tea polyphenol aqueous solution at a flow rate of 0.4 times the column volume / hour. Elution at flow rate per hour, and then elution with 4 times column volume of 20% ethanol, according to the detection results of silica gel thin layer chromatography, collect the epigallocatechin gallate flow fraction, and the epigallocatechin gallate flow fraction Concentrate under reduced pressure below 45°C to a small volume (1500mL) to obtain sample B. Sample B is placed on a 2kg polyamide (80-100 mesh) ...

Embodiment 2

[0026] Weigh 100 grams of a tea polyphenol extract with an epigallocatechin gallate content of 50%, and add 400 mL of deionized water to dissolve it. Get 1.0kg of D101 macroporous adsorption resin and pack it into the column with 95% ethanol, wash off the porogen and polymerized monomer residues in the resin with 95% ethanol until the eluent is concentrated and qualified by gas phase detection, and then use Replace the ethanol in the column with deionized water, and load the above tea polyphenol aqueous solution at a flow rate of 0.3 times the column volume / hour. After the sample enters the column, first use 3 times the column volume of deionized water to Flow rate elution, and then use 4 times column volume 40% ethanol to 0.3 times column volume / hour flow rate elution, according to the silica gel thin layer chromatography detection result, collect epigallocatechin gallate fraction, reduce at 45 ℃ Concentrate under pressure to a small volume (600mL) to obtain sample B. Sample ...

Embodiment 3

[0029] The epigallocatechin gallate, epicatechin and epigallocatechin prepared in the present invention are subjected to in vitro α-glucosidase and α-amylase inhibition tests.

[0030] α-glucosidase activity assay

[0031] Use p-nitrophenol-α-D-glucopyranoside (4-Nitrophenylα-D-glucopyranoside, PNPG) as the substrate. The reaction system was: 67mmol / L potassium phosphate buffer (pH6.8) 2ml, 1mg / ml reduced glutathione 50μl, 0.57U / ml α-glucosidase (sigma company, Type I: fromBakers Yeast) 100μl, 37 After incubating at ℃ for 10min, add 20mmol / L PNPG 200μl, react at 37℃ for 30min, add 0.1mol / L Na 2 CO 3 Stop solution 10ml. Acarbose was used as a positive control. Absorbance was measured at 400 nm.

[0032]

[0033] In the formula:

[0034] A 空白 : Absorption value after reaction without adding sample

[0035] A 样品 : Absorption value after adding sample to react

[0036] A 背景 : Only add the absorbance value of the sample

[0037] The results of α-glucosidase activity as...

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Abstract

The invention discloses a separation and purification method of main catechin components in tea polyphenol and glycosidase activity thereof. In the separation and purification method, tea polyphenol extracts are absorbed by macroporous adsorption resin of nonpolar or weak polar polystyrene; water and ethanol are used for the elution; alcohol eluates, through polyamide columns for the chromatography and are eluted respectively by water and ethanol. The water recrystallization is carried out on the alcohol eluates of the polyamide columns to obtain EGCG pure products with the purity larger than 95 percent and the yield larger than 50 percent; reversed-phase C18 filled columns are used for the chromatography of water eluates of the polyamide columns to obtain EC and EGC; the ethanol of 50-80 percent is used for the recrystallization to obtain pure products of the EC and EGC with the purity all larger than 96 percent. Tests in vitro show that EGCG, EGC and EC all have inhibitory activity on alpha-glucosidase and alpha-amylase and can be used for preparing weight reduction products reducing the absorption of carbohydrates or drugs or health products lowering the postprandial blood glucose.

Description

technical field [0001] The invention relates to a method for separating and purifying catechin components in tea polyphenols and uses of the obtained monomers. In particular, it relates to the separation and purification method of three important catechin components of epigallocatechin gallate (EGCG), epicatechin (EC) and epigallocatechin (EGC) in tea polyphenols and the method for the three important The inhibitory activity of catechin components alpha-glucosidase and alpha-amylase can be used to prepare weight-loss products for reducing carbohydrate absorption or medicines or health products for reducing postprandial blood sugar. Background technique [0002] Epigallocatechin gallate (EGCG), epicatechin (EC), and epigallocatechin (EGC) are the catechins with the highest content in tea and are also important active ingredients. They have anti-tumor, Lowering blood fat, anti-oxidation, anti-aging, anti-HIV, anti-cancer metastasis, anti-fibrosis, antibacterial, anti-fungal, ...

Claims

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Application Information

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IPC IPC(8): C07D311/62A61K31/353A61P3/04A61P3/10
Inventor 宋纯清叶晓平茅仁刚
Owner SHANGHAI NORMAL UNIVERSITY
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