Method for preparing high-purity theaflavin composite

A technology of theaflavin and compound, which is applied in the field of preparation of high-purity theaflavin compound, can solve the problems of low product purity, high cost, unstable properties, etc., and achieves high product yield, environmental protection and simple process Effect

Inactive Publication Date: 2012-07-25
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the chemical research of theaflavins cannot meet the requirements of the research in the field of medicine. For example, the purification of theaflavins is difficult and its properties are not stable, which limits the in-depth medicinal research.
Therefore, so far, the industrialization technology of theaflavin extraction is still immature, which is characterized by low product purity, high cost, and low production capacity, which seriously hinders the research on natural medicines and functional foods using theaflavins as raw materials. and development

Method used

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  • Method for preparing high-purity theaflavin composite
  • Method for preparing high-purity theaflavin composite

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Dissolve 4.00g of catechin mixture (EGC>20%, EC>9%, EGCG>10%) in 400ml polyphenol oxidase crude enzyme liquid (add 100ml lemon pH 5.5 to the polyphenol oxidase crude enzyme liquid Acid-disodium hydrogen phosphate buffer solution), feed oxygen, it is advisable that the bubbles do not overflow the reaction vessel, stir, and react in a constant temperature water bath at 30°C for 30min, and immediately pre-cool to -18 Ethyl acetate extraction at ~-20°C, the ester layer was concentrated in vacuo at 40°C, the ethyl acetate was recovered, and dried to obtain 1.70 g of crude theaflavin; the crude theaflavin was dissolved in 20 ml of 20% ethanol aqueous solution, The solution is passed through a chromatographic column (φ3cm×60cm, 30-60 mesh, pretreated according to the general method of polyamide treatment for column chromatography) equipped with polyamide powder at a flow rate of 0.6-0.8BV / h until the adsorption is saturated, followed by Use 2.5BV of 20% volume concentration of...

Embodiment 2

[0025] Dissolve 8.00g of small leaf catechin mixture (EGC>10%, ECG>9%, EGCG>40%) in 800ml polyphenol oxidase crude enzyme solution, feed oxygen, it is advisable that bubbles do not overflow the reaction vessel, stir , reacted in a constant temperature water bath at 20°C for 40min, immediately extracted with ethyl acetate pre-cooled to -18~-20°C equal to the volume of catechin mixture solution, concentrated the ester layer in vacuum at 45°C, recovered ethyl acetate, and dried to obtain Crude theaflavin 3.76g. This theaflavin crude product is dissolved with the ethanol aqueous solution of 40ml 20% volume concentration, and dissolving solution is passed through the chromatographic column (φ 3cm * 60cm that NKA macroporous adsorption resin is housed with the flow velocity of 0.6~0.8BV / h, according to column chromatography Use the macroporous adsorption resin treatment general method for pretreatment) to adsorption saturation, and then use 3BV 20% volume concentration of ethanol aq...

Embodiment 3

[0027] Dissolve 8.00g of catechin mixture (EGC>20%, EC>9%, EGCG>10%) in 800ml polyphenol oxidase crude enzyme solution, feed oxygen, it is advisable that bubbles do not overflow the reaction vessel, stir, React in a constant temperature water bath at 40°C for 20 minutes, immediately extract with an equal volume of ethyl acetate pre-cooled to -18~-20°C with the catechin mixture solution, and concentrate the ester layer in vacuum at 42°C, recover ethyl acetate, and dry to obtain Crude theaflavin 3.60g. This theaflavin crude product is dissolved with 50ml of 20% ethanol aqueous solution of volume concentration, and the dissolving liquid is passed through the chromatographic column (φ 3cm * 60cm, 30~60 meshes of polyamide powder with the flow rate of 0.6~0.8BV / h, according to The column chromatography is pretreated by the general method of polyamide treatment) to adsorption saturation, and then eluted with 2BV of ethanol aqueous solution with a volume concentration of 20% and 2.5B...

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Abstract

The invention provides a method for preparing a high-purity theaflavin composite. The method comprises the following steps of: adding a mixture of green tea and catechinic acid into a crude polyphenol oxidase liquid for carrying out in-vitro oxidation; then, extracting the mixture with ethyl acetate; concentrating the extract in vacuum and drying to obtain a theaflavin crude product; dissolving the theaflavin crude product with ethanol water with volume concentration being 20% and then absorbing by using a chromatographic column with absorbing resin till the chromatographic column with absorbing resin is in a saturated state; eluting by using the ethanol water with volume concentration being 20%, and then eluting by using acetone or ethanol water with volume concentration being 60%; concentrating the eluting liquid, namely the collected acetone or ethanol water solution with volume concentration being 60% in vacuum; and obtaining the high-purity theaflavin composite through freeze drying. The method provided by the invention has the advantages of simple process and low cost and avoids using toxic and harmful organic reagents during a production process, thereby being environment-friendly. Furthermore, the high-purity theaflavin composite with different theaflavin components can be obtained by controlling different reaction conditions, so that the product yield is high and the content is high.

Description

Technical field: [0001] The invention relates to a preparation method of high-purity theaflavin complex. Background technique: [0002] Theaflavin refers to the orange-yellow substance dissolved in ethyl acetate in black tea, which is formed by oxidative condensation of tea polyphenols (mainly catechins and their derivatives) in the presence of polyphenol oxidase. The content of theaflavins in black tea is about 0.5% to 2.0%, but theaflavins are not only an important quality component of black tea, but also have many potential effects related to human health, such as anti-oxidation, anti-aging, anti-mutation, anti Cancer prevention, lowering blood sugar, lowering blood fat, preventing and treating cardiovascular diseases, etc., are even superior to catechins in some functions. Theaflavin is the first time to find a compound with definite pharmacological effects from tea; after clinical trials, it has been verified that theaflavin has the effect of regulating blood lipids an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/16C07D311/62
Inventor 王坤波刘仲华黄建安刘芳龚雨顺
Owner HUNAN AGRICULTURAL UNIV
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