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Method for extracting tricin from alfalfa

A technology of alfalfa and alfalfa, applied in the biological field, can solve the problems of cumbersome operation, serious sample loss, poor reproducibility, etc., and achieve the effect of solving serious pollution, large amount of preparation, and short preparation cycle

Inactive Publication Date: 2013-02-06
苏州宝泽堂医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This separation method has serious sample loss, tedious operation and poor reproducibility

Method used

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  • Method for extracting tricin from alfalfa

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Dry and pulverize alfalfa grass, take 1kg and add 8 times the amount of 80% ethanol to reflux for extraction twice, concentrate the extract to no alcohol, add 200ml polyamide resin column for adsorption, take 5BV40% ethanol solution to elute impurities, and then use 70% ethanol solution Elution, TLC detection, collection of high-concentration fractions and concentration, the concentrated solution was extracted three times with an equal volume of ethyl acetate, and the ethyl acetate was recovered to obtain 8 g of crude product. Take n-hexane, ethyl acetate, methanol, and water and mix them according to 5:7:4:6. After standing and stratifying, take the upper phase and fill the high-speed countercurrent chromatography column, turn the host, adjust the speed to 700rpm, and pump into the lower phase for flow. Phase, after the dynamic equilibrium is established, the flow rate is adjusted to 3ml / min, and the crude product dissolved in the mobile phase is injected by the injecti...

Embodiment 2

[0018] Dry and pulverize alfalfa grass, take 1kg and add 5 times the amount of 60% ethanol to reflux and extract for 3 times, concentrate the extract to no alcohol, add 250ml polyamide resin column for adsorption, take 4BV50% ethanol solution to elute impurities, and then use 70% ethanol solution Elution, TLC detection, collection of high-concentration fractions and concentration, the concentrated solution was extracted three times with an equal volume of ethyl acetate, and the ethyl acetate was recovered to obtain 10 g of crude product. Take n-hexane, ethyl acetate, methanol, and water and mix them according to 4:5:3:4. After standing and stratifying, take the upper phase and fill the high-speed countercurrent chromatography column, turn the host, adjust the speed to 800rpm, and pump into the lower phase for flow. Phase, after the dynamic equilibrium is established, the flow rate is adjusted to 2ml / min. At the same time, the crude product dissolved in the mobile phase is injec...

Embodiment 3

[0020] Dry and pulverize alfalfa grass, take 1kg and add 7 times the amount of 70% ethanol to reflux for extraction twice, concentrate the extract to no alcohol, add 200ml polyamide resin column for adsorption, take 4BV50% ethanol solution to elute impurities, and then use 70% ethanol solution Elution, TLC detection, collection of high-concentration fractions and concentration, the concentrated solution was extracted three times with an equal volume of ethyl acetate, and the ethyl acetate was recovered to obtain 7 g of crude product. Mix n-hexane, ethyl acetate, methanol, and water at a ratio of 7:8:5:6, and after standing for stratification, take the upper phase and fill the high-speed countercurrent chromatography column, turn the host, adjust the speed to 700rpm, and pump into the lower phase for flow Phase, after the establishment of dynamic equilibrium, the flow rate was adjusted to 1ml / min, and the crude product dissolved in the mobile phase was injected by the injection ...

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Abstract

The invention belongs to the technical field of biology, and discloses a method for extracting tricin from alfalfa. The method is characterized in comprising the steps that: (1) alfalfa is crushed; 5-8 time amount of 60-80% ethanol is added for carrying out reflux extraction 2-3 times; an obtained extraction liquid is concentrated until no alcohol is left; the extraction liquid is absorbed by using polyamide resin column; an ethanol solution is used in gradient elution; thin-layer detection is carried out; high-concentration fraction is collected and concentrated; ethyl acetate is added for extracting; and ethyl acetate is recovered, such that a crude product is obtained; (2) n-hexane, ethyl acetate, methanol, and water are mixed; an upper-phase is used for filling a high-speed countercurrent chromatographic column; a main machine is rotated, and a lower-phase is pumped in as a mobile phase; the crude product dissolved by the mobile phase is fed by using a feeding valve, and is subjected to UV detector online monitoring; a target component is collected; obtained fractions are combined; and the mixture is subjected to reduced-pressure drying, such that high-purity tricin is obtained. The method provided by the invention has the advantages of short-period, simple operation, large preparation amount, high yield, and the like. The method is suitable for preparations of high-purity tricin.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for extracting alfalfa from alfalfa. Background technique [0002] Alfalfa is also called wheat flavin, wheat flavone, chemical name 5,7-dihydroxy-2-(4-hydroxy-3,5-dimethoxyphenyl)benzopyran-4-one, CAS No. 520 -32-1, melting point 285-286°C, molecular formula C17H1407, molecular weight 330, molecular structural formula [0003] [0004] Alfalfa is a kind of flavonoid, which has slight antioxidant effect, anti-smooth muscle relaxation and anti-tumor effect, and also has dyeing function. [0005] Alfalfa widely exists in plants, but the content is low, not enough for separation and purification, and the content is more in alfalfa. The method for preparing high-purity alfalfalin mostly adopts solid-packed column methods, such as silica gel column, reversed-phase silica gel column and gel column method. This separation method suffers from severe sample loss, cumbersome ope...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D311/30C07D311/40
Inventor 李法庆刘东锋
Owner 苏州宝泽堂医药科技有限公司
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