CN101538308A

CN101538308A - Method for extracting and preparing high-purity ginsenoside Re from herminium by high speed counter current chromatography

Info

Publication number: CN101538308A
Application number: CN200810050528A
Authority: CN
Inventors: 孙成贺; 王英平; 李范英; 赵景辉
Original assignee: Individual
Current assignee: Individual
Priority date: 2008-03-21
Filing date: 2008-03-21
Publication date: 2009-09-23
Anticipated expiration: 2028-03-21
Also published as: CN101538308B

Abstract

The invention relates to a method for extracting and preparing high-purity ginsenoside Re from herminium by high speed counter current chromatography, which is characterized by comprising the following steps: soaking fruit residues of the herminium in 70 percent ethanol solution through overnight; performing ultrasonic extraction for 30 minutes for three times repeatedly and concentrating extract liquid after filtration to obtain herminium coarse extract; and selecting ethyl acetate 0.5-2, butanol 4-7 and water 6-9 according to a volume ratio to form a solvent system and separating at a room temperature of from 16 to 28 by using a high speed counter current chromatographic instrument. The method for preparing and separating ginsenoside Re in large scale avoids the use of solid filler as well as irreversible adsorption, sample loss and pollution, and is high in efficiency, quick and low in cost. The obtained ginsenoside Re has a purity of more than 98 percent, and the method is suitable for separating and preparing the insenoside Re from herminium coarse extracts prepared under various process conditions.

Description

From Herba Herminii, separate the method for preparing the high purity ginsenoside Re with high-speed countercurrent chromatography

Technical field:

The present invention relates to a kind ofly from Herba Herminii, separate the method for preparing the high purity ginsenoside Re, can be widely used in the separation and purification of active substance in the fields such as biology, pharmacy, environmental protection and agricultural with high-speed countercurrent chromatography.

Background technology:

Herba Herminii is the mature fruit of Araliaceae genseng (Panax ginseng C.A.Mey).Contain number of chemical compositions such as saponin(e, polysaccharide, alkaloid, sterol, volatile oil, amino acid, inorganic elements in the Herba Herminii.Pharmaceutical research shows that it has lowering blood glucose, antishock, protection cardiac muscle, renal function protecting, strengthening immunity and delays senility, improves multiple pharmacological effect such as memory capability.Saponin content is abundant in the Herba Herminii, and is wherein the highest with Re content again, and the ginsenoside Re is by " Chinese pharmacopoeia is defined as the quality control composition of panax species and products thereof, still is the significant composition of genseng and goods finger printing thereof simultaneously.

In the past Bao Dao ginsenoside Re to separate preparation mainly be to adopt column chromatography, need to use solid packing, easily produce irreversible adsorption, have that sample easily loses, easily pollution, poor efficiency, a shortcoming such as speed is slow, cost is high and separating step is loaded down with trivial details.

Summary of the invention:

The object of the present invention is to provide and a kind ofly from Herba Herminii, separate the method for preparing the high purity ginsenoside Re with high-speed countercurrent chromatography, high-speed countercurrent chromatography need not use solid packing, carry out the separation and purification of natural product under the mode of solvent systems high speed planetary motion in supporting tube that the application two-phase is immiscible, thereby do not have irreversible adsorption, have sample free of losses, the isolating advantage of pollution-free, efficient, quick and big preparation amount.

Technical scheme of the present invention is achieved in that with high-speed countercurrent chromatography separates the method for preparing the high purity ginsenoside Re from Herba Herminii, it is characterized in that: adopt a small amount of preparation of analysis mode high-speed counter-current chromatograph and the relatively large preparation of half countercurrent chromatography instrument; Its technological process is as follows:

The pomace of Herba Herminii is spent the night with 70% alcohol solution dipping, and supersound extraction 30min extracts three times repeatedly, concentrate behind the extracting liquid filtering the Herba Herminii crude extract;

Select for use ethyl acetate 0.5-2, propyl carbinol 4-7, water 6-9 for the solvent systems system places separating funnel by following volume ratio, shake up the back static layering; Ready to balance separates upper and lower phase after for some time, more than be stationary phase mutually, is moving phase mutually down; It is stand-by in being dissolved under the 2ml mutually to get the Herba Herminii crude extract; Separate with high-speed counter-current chromatograph under 16-28 ℃ of condition in room temperature, stationary phase is filled with chromatography column, the opening speed controller, when reaching desired speed, beginning pumps into moving phase with certain flow rate, when moving phase flows out, by the six-way injection valve sample introduction; By head end wash-out caudad, main frame just changes, and rotating speed is 1300r/min, regularly collects component.

Positively effect of the present invention is: provide a kind of and need not use solid packing, there is not irreversible adsorption, the method that sample free of losses, pollution-free, efficient, quick, low-cost and big preparation amount separate the ginsenoside Re, the ginsenoside Re's purity that obtains can reach more than 98%, is applicable to separate the preparation ginsenoside Re from the Herba Herminii crude extract of kinds of processes condition preparation.

Embodiment:

The present invention will be further described below in conjunction with embodiment:

Embodiment 1:

Select for use ethyl acetate 0.5, propyl carbinol 4, water 6 for the solvent systems system places separating funnel by following volume ratio, shake up the back static layering; Ready to balance separates upper and lower phase after for some time, more than be stationary phase mutually, is moving phase mutually down; It is stand-by in being dissolved under the 2ml mutually to get the Herba Herminii crude extract; Under 16-28 ℃ of condition, produce CCC-3000 type high-speed counter-current chromatograph in room temperature and separate, stationary phase is filled with chromatography column, the opening speed controller with the U.S., when reaching desired speed, beginning pumps into moving phase with certain flow rate, when moving phase flows out, by the six-way injection valve sample introduction; By head end wash-out caudad, main frame just changes, and rotating speed is 1300r/min, regularly collects component.

Embodiment 2:

Select for use ethyl acetate 2, propyl carbinol 7, water 9 for the solvent systems system places separating funnel by following volume ratio, shake up the back static layering; Ready to balance separates upper and lower phase after for some time, more than be stationary phase mutually, is moving phase mutually down; It is stand-by in being dissolved under the 10ml mutually to get the Herba Herminii crude extract; Under 16-28 ℃ of condition, produce CCC-3000 type high-speed counter-current chromatograph in room temperature and separate, stationary phase is filled with chromatography column, the opening speed controller with the U.S., when reaching desired speed, beginning pumps into moving phase with certain flow rate, when moving phase flows out, by the six-way injection valve sample introduction; By head end wash-out caudad, main frame just changes, and rotating speed is 900r/min, regularly collects component.

Embodiment 3:

Select for use ethyl acetate 1, propyl carbinol 6, water 7 for the solvent systems system places separating funnel by following volume ratio, shake up the back static layering; Ready to balance separates upper and lower phase after for some time, more than be stationary phase mutually, is moving phase mutually down; It is stand-by in being dissolved under the 10ml mutually to get the Herba Herminii crude extract; Under 16-28 ℃ of condition, produce CCC-3000 type high-speed counter-current chromatograph in room temperature and separate, stationary phase is filled with chromatography column, the opening speed controller with the U.S., when reaching desired speed, beginning pumps into moving phase with certain flow rate, when moving phase flows out, by the six-way injection valve sample introduction; By head end wash-out caudad, main frame just changes, and rotating speed is 900r/min, regularly collects component.

Claims

1, from Herba Herminii, separates the method for preparing the high purity ginsenoside Re with high-speed countercurrent chromatography, it is characterized in that: adopt a small amount of preparation of analysis mode high-speed counter-current chromatograph and the relatively large preparation of half countercurrent chromatography instrument; Its technological process is as follows:

Select for use ethyl acetate 0.5-2, propyl carbinol 4-7, water 6-9 for the solvent systems system places separating funnel by following volume ratio, shake up the back static layering; Ready to balance separates upper and lower phase after for some time, more than be stationary phase mutually, is moving phase mutually down; It is stand-by in being dissolved under the 2ml-10ml mutually to get the Herba Herminii crude extract; Separate with high-speed counter-current chromatograph under 16-28 ℃ of condition in room temperature, stationary phase is filled with chromatography column, the opening speed controller, when reaching desired speed, beginning pumps into moving phase with certain flow rate, when moving phase flows out, by the six-way injection valve sample introduction; By head end wash-out caudad, main frame just changes, and rotating speed is 900-1300r/min, regularly collects component.