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Method for separating active ingredients from aquilaria sinensis lamina by utilizing high-speed countercurrent chromatography

A high-speed countercurrent chromatography, active ingredient technology, applied in the field of separation of anticancer and antioxidant active ingredients from leaves

Active Publication Date: 2013-06-12
北京利科德科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

HSCCC technology is a liquid-liquid partition chromatography technology without any solid support, and there will be no irreversible adsorption of sample components. It is more effective and economical for flavonoids and other substances that are easily adsorbed and lost during solid-liquid chromatography. Using HSCCC technology to separate and purify the active components of A. argentina leaves, there has been no report at home and abroad.

Method used

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  • Method for separating active ingredients from aquilaria sinensis lamina by utilizing high-speed countercurrent chromatography
  • Method for separating active ingredients from aquilaria sinensis lamina by utilizing high-speed countercurrent chromatography
  • Method for separating active ingredients from aquilaria sinensis lamina by utilizing high-speed countercurrent chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] The active ingredient A was isolated from petroleum ether part extract (sample I):

[0069] 1. Prepare 1000mL of n-hexane:ethyl acetate:methanol:water (V / V)=1:1.5:1.5:1 solvent system, fully shake and mix, then stand and separate to obtain upper and lower phase solutions, and then The upper and lower phases were separated and ultrasonically degassed for 30 minutes before use;

[0070] 2. The upper phase solution is used as the mobile phase, and the lower phase solution is the stationary phase. Turn on the injection pump to pump the stationary phase into the maximum flow rate of the equipment and fill the countercurrent chromatographic column, and then turn on the high-speed countercurrent chromatographic host, detector, and chromatographic workstation. Pump the mobile phase at a speed of 815rpm and a flow rate of 1.0ml / min;

[0071] 3. Take 50.0mg of sample I and dissolve it in 20ml lower phase in a test tube until completely dissolved;

[0072] 4. After the curve of ...

Embodiment 2

[0103] The active ingredients A, B, and C are separated by countercurrent chromatographic separation using the extract of ethyl acetate, and the solvent system is firstly n-hexane:ethyl acetate:methanol:water (V / V)=1.5:2:2:1.5 Isolate the active component A by the separation method of petroleum ether extract in Example 1, then use silica gel column chromatography to roughly separate the ethyl acetate extract to obtain the crude extract containing the target component, and finally use normal hexane: ethyl acetate: methanol: Water (V / V)=0.5:1:1:0.5 is the solvent system to separate the active components B and C.

[0104] Wherein the parameter of high-speed countercurrent chromatographic separation active component A is:

[0105] The high-speed countercurrent chromatography host rotates clockwise at a speed of 900rpm; the sample volume is 60.0mg, the retention value of the stationary phase is 73.0%, and the flow rate of the mobile phase is 1.2ml / min; the detection wavelength is 2...

Embodiment 3

[0112] The active ingredient A is separated from petroleum ether extract and ethyl acetate extract by high-speed countercurrent chromatography, and its parameters are:

[0113] The main engine of high-speed countercurrent chromatography rotates clockwise at 750rpm; the sample volume is 40.0mg, the retention value of the stationary phase is 65.4%, the flow rate of the mobile phase is 0.8ml / min; the detection wavelength is 254nm; the separation temperature is 17~23°C;

[0114] Then the ethyl acetate extract is roughly separated by silica gel column chromatography, and the fraction containing the target component is retained as a crude extract, and finally the crude extract is subjected to high-speed countercurrent chromatography to obtain active ingredient B and active ingredient C. Wherein the separation parameters of high-speed countercurrent chromatography are:

[0115] The main engine of high-speed countercurrent chromatography rotates clockwise at 750rpm; the sample volume ...

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Abstract

The invention discloses a method for separating active ingredients from an aquilaria sinensis lamina by utilizing a high-speed counter current chromatography. The method comprises the following steps of: extracting the dried and smashed aquilaria sinensis lamina by virtue of a percolation extract method at the room temperature to obtain an aquilaria sinensis lamina extracting solution extractum; extracting the extracting solution extractum for 3-4 times by sequentially utilizing petroleum ether,and ethyl acetate, respectively decompressing and concentrating a petroleum ether extract liquor and an ethyl acetate extract liquor to extractums; then respectively separating the petroleum ether part extractum and the ethyl acetate part extractum to obtain an active ingredient A of 7,4'-dimethoxy-5-hydroxy flavone by utilizing the high-speed counter current chromatography; and then separating the ethyl acetate part extractum through high-speed countercurrent chromatography to obtain an active ingredient B of 3,5,7,3',4'-pentahydroxy-flavone and an active ingredient C of 3,5,7,4'-tetrahydroxy-flavone after coarsely separating the ethyl acetate part extractum by virtue of a column chromatography on silica gel. The active ingredients separated by the method are low in loss, high in separation speed, high in purity, stable and easy to apply.

Description

【Technical field】 [0001] The present invention relates to a method for separating the active ingredients in the leaves of A. chinensis, the active ingredients are anti-cancer active ingredients and anti-oxidation active ingredients, in particular to a method for separating the active ingredients in the leaves of A. Approach to anticancer and antioxidant active ingredients in leaves. 【Background technique】 [0002] Domestic agarwood is the resinous wood of Aquilaria sinensis (Lour.) Gilg, a plant of the genus Aquilaria sinensis (Lour.) Gilg. Akiras sinensis is the source plant of domestic Agarwood medicinal materials, and the active components of its leaves are reported to have activities such as antioxidant, analgesic, anti-inflammatory, regulating diabetes, and inhibiting respiratory diseases. [0003] As we all know, cancer is a medical problem worldwide today. Wang Honggang et al. [Research on the Antitumor Active Chemical Components of Agarwood Leaves, Forest Product C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D311/40C07D311/30A61P35/00
Inventor 杨懋勋陈河如梁耀光张天佑谢朝良蒋建平高慎凎李小玉
Owner 北京利科德科技有限公司
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