CN103254208A

CN103254208A - Extraction and purification method of amorphin

Info

Publication number: CN103254208A
Application number: CN201310197309XA
Authority: CN
Inventors: 苏刘花; 万冬梅
Original assignee: Nanjing Zelang Agricultural Development Co Ltd
Current assignee: Nanjing Zelang Agricultural Development Co Ltd
Priority date: 2013-05-24
Filing date: 2013-05-24
Publication date: 2013-08-21

Abstract

The invention discloses an extraction and purification method of amorphin. The method comprises the following steps of: crushing fruits of amorpha fruticosa, adding into a supercritical extraction kettle, performing supercritical CO2 extraction under the conditions that the extraction pressure is 28-40MPa and the temperature is 45-60 DEG C, resolving to obtain an extract, adsorbing through a polyamide resin column, eluting with alcohol water, collecting eluent, concentrating and drying to obtain a crude product, performing high-speed counter-current chromatographic separation and purification on the crude product, taking n-hexane-ethyl acetate-methanol-water as a solvent system, detecting through an ultraviolet detector, collecting fractions, recovering a reagent, and drying to obtain the amorphin. The method for extracting and purifying the amorphin has the advantages of simple operation, high efficiency and high product purity, and is suitable for industrial preparation.

Description

A kind of extracting and purifying method of amorphin

Technical field

The invention belongs to the extractive technique field of crude substance, relate to a kind of extracting and purifying method of amorphin.

Background technology

Amorphin (Amorphin), molecular formula are C ₃₄H ₄₀O ₁₆, molecular weight is 704.68, mp.154-155 ℃, is present in pulse family (Laguminosae) the false indigo Amorpha fruticosaL. fruit.Amorphin can suppress phosphodiesterase, and the rat heart phosphodiesterase is had remarkable restraining effect, and IC50 is active luteolin, kaempferol, the Quercetins of surpassing of 1.7 μ M; Can suppress the super effect of coagulating, arteriosclerotic super the coagulating property of rabbit experiment is had restraining effect; Have hepatoprotective effect, rat is had provide protection by the hepatic injury that CCl4 or D-galactosamine cause.

Summary of the invention

The extracting and purifying method that the purpose of this invention is to provide a kind of amorphin.

For achieving the above object, the present invention is by the following technical solutions:

A kind of extracting and purifying method of amorphin is characterized in that may further comprise the steps: the fruit of false indigo being pulverized, added in the supercritical extraction reactor, is entrainment agent with methyl alcohol, carries out supercritical CO ₂Extraction is resolved, and gets extract, use dissolve with methanol, add the polyamide resin drying, the dress post, the alcohol solution wash-out, collect elutriant, concentrate drying gets crude product, with crude product through the high speed adverse current chromatogram separation and purification, normal hexane-ethyl acetate-methanol-water is solvent systems, UV-detector detects, and collects flow point and reclaims reagent, is drying to obtain.

Described supercritical CO ₂Extraction temperature is 45-60 ℃, and extracting pressure is 28-40MPa, and resolution temperature is 35-48 ℃, and parsing pressure is 6-8.5MPa, extraction time 40-120min, CO ₂Flow is 3.5-5ml/g crude drug/min.

Described alcohol solution is the aqueous ethanolic solution of 20-80%.

Described high speed adverse current chromatogram separation and purification, the volume ratio of solvent systems normal hexane-ethyl acetate-methanol-water are (2-5): (1-2.5): (1-2.5): (3-5).

The invention has the beneficial effects as follows: (1) supercritical fluid extraction, efficient height, extraction yield height; (2) adopt the high speed adverse current chromatogram separation and purification, the product-free loss, separation is efficient, preparation amount is big, separation system is reusable, and environmental pollution is little.

Further specify the present invention below in conjunction with embodiment, but the scope of protection of present invention is not limited to following embodiment.

Embodiment

Embodiment 1:

The fruit of false indigo being pulverized 20 mesh sieves, get 10kg and add in the supercritical extraction reactor, is entrainment agent with methyl alcohol, and setting extraction temperature is 60 ℃, and extracting pressure is 35MPa, and resolution temperature is 42 ℃, and parsing pressure is 7.5MPa, extraction time, CO ₂Flow is 4ml/g crude drug/min, and extraction 80min resolves, and gets extract, use dissolve with methanol, add polyamide resin and mix sample, drying, dress post, with 4BV20% ethanol elution impurity, use the 75% ethanol elution effective constituent of 6BV more earlier, collect effective constituent concentrating under reduced pressure, the dry crude product that gets.Get the 4:2.5:2.5:3 mixing by volume of normal hexane, ethyl acetate, methyl alcohol, water, after the layering, getting the phase that fixes mutually, is moving phase down mutually, the adjustment rotating speed is 850rpm, pump into down back control flow velocity 2ml/min mutually, take off the phased soln crude product, by the sampling valve sample introduction, collect the amorphin flow point, reclaim reagent, be drying to obtain the amorphin product, content is 97.6%.

Embodiment 2:

The fruit of false indigo being pulverized 20 mesh sieves, get 10kg and add in the supercritical extraction reactor, is entrainment agent with methyl alcohol, and setting extraction temperature is 50 ℃, and extracting pressure is 28MPa, and resolution temperature is 40 ℃, and parsing pressure is 6.5MPa, CO ₂Flow is 3.5ml/g crude drug/min, and extraction 100min resolves, and gets extract, use dissolve with methanol, add polyamide resin and mix sample, drying, dress post, with 3BV30% ethanol elution impurity, use the 75% ethanol elution effective constituent of 5BV more earlier, collect effective constituent concentrating under reduced pressure, the dry crude product that gets.Get the 3:2:2:4 mixing by volume of normal hexane, ethyl acetate, methyl alcohol, water, after the layering, getting the phase that fixes mutually, is moving phase down mutually, the adjustment rotating speed is 900rpm, pump into down back control flow velocity 2ml/min mutually, take off the phased soln crude product, by the sampling valve sample introduction, collect the amorphin flow point, reclaim reagent, be drying to obtain the amorphin product, content is 97.1%.

Embodiment 3:

The fruit of false indigo being pulverized 20 mesh sieves, get 10kg and add in the supercritical extraction reactor, is entrainment agent with methyl alcohol, and setting extraction temperature is 45 ℃, and extracting pressure is 40MPa, and resolution temperature is 48 ℃, and parsing pressure is 6MPa, CO ₂Flow is 3.5ml/g crude drug/min, and extraction 120min resolves, and gets extract, use dissolve with methanol, add polyamide resin and mix sample, drying, dress post, with 4BV35% ethanol elution impurity, use the 80% ethanol elution effective constituent of 5BV more earlier, collect effective constituent concentrating under reduced pressure, the dry crude product that gets.Get the 2:1:1:3 mixing by volume of normal hexane, ethyl acetate, methyl alcohol, water, after the layering, getting the phase that fixes mutually, is moving phase down mutually, the adjustment rotating speed is 750rpm, pump into down back control flow velocity 3ml/min mutually, take off the phased soln crude product, by the sampling valve sample introduction, collect the amorphin flow point, reclaim reagent, be drying to obtain the amorphin product, content is 98.0%.

Embodiment 4:

The fruit of false indigo being pulverized 20 mesh sieves, get 10kg and add in the supercritical extraction reactor, is entrainment agent with methyl alcohol, and setting extraction temperature is 60 ℃, and extracting pressure is 40MPa, and resolution temperature is 35 ℃, and parsing pressure is 8.5MPa, CO ₂Flow is 5ml/g crude drug/min, and extraction 40min resolves, and gets extract, use dissolve with methanol, add polyamide resin and mix sample, drying, dress post, with 3BV30% ethanol elution impurity, use the 80% ethanol elution effective constituent of 4BV more earlier, collect effective constituent concentrating under reduced pressure, the dry crude product that gets.Get the 3:1.5:1.5:5 mixing by volume of normal hexane, ethyl acetate, methyl alcohol, water, after the layering, getting the phase that fixes mutually, is moving phase down mutually, the adjustment rotating speed is 800rpm, pump into down back control flow velocity 2.5ml/min mutually, take off the phased soln crude product, by the sampling valve sample introduction, collect the amorphin flow point, reclaim reagent, be drying to obtain the amorphin product, content is 97.2%.

Claims

1. the extracting and purifying method of an amorphin is characterized in that may further comprise the steps: with the fruit pulverizing of false indigo, in the adding supercritical extraction reactor, be entrainment agent with methyl alcohol, carry out supercritical CO ₂Extraction is resolved, and gets extract, use dissolve with methanol, add the polyamide resin drying, the dress post, the alcohol solution wash-out, collect elutriant, concentrate drying gets crude product, with crude product through the high speed adverse current chromatogram separation and purification, normal hexane-ethyl acetate-methanol-water is solvent systems, UV-detector detects, and collects flow point and reclaims reagent, is drying to obtain.

2. the extracting and purifying method of a kind of amorphin as claimed in claim 1 is characterized in that described supercritical CO ₂Extraction temperature is 45-60 ℃, and extracting pressure is 28-40MPa, and resolution temperature is 35-48 ℃, and parsing pressure is 6-8.5MPa, extraction time 40-120min, CO ₂Flow is 3.5-5ml/g crude drug/min.

3. the extracting and purifying method of a kind of amorphin as claimed in claim 1 is characterized in that described alcohol solution is the aqueous ethanolic solution of 20-80%.

4. the extracting and purifying method of a kind of amorphin as claimed in claim 1, it is characterized in that described high speed adverse current chromatogram separation and purification, the volume ratio of solvent systems normal hexane-ethyl acetate-methanol-water is (2-5): (1-2.5): (1-2.5): (3-5).