CN101773740A

CN101773740A - Organic matter-water-organic matter purification, separation and leaching technology

Info

Publication number: CN101773740A
Application number: CN200910102415A
Authority: CN
Inventors: 陈锡武
Original assignee: Individual
Current assignee: Individual
Priority date: 2009-01-14
Filing date: 2009-01-14
Publication date: 2010-07-14

Abstract

The invention relates to an organic matter-water-organic matter purification, separation and leaching technology. After two organic matters with significantly different hydrophilic nature are mixed with water by a certain ratio, the solution becomes a two-phase or a multi-phase system; and according to the distribution coefficient difference of the target matter in the system, so leaching and separation technology for purification, separation and target matter extraction is realized.

Description

Organic matter-water-matter-water-organic matter purification, separation and leaching technology

Affiliated technical field

The present invention relates to extraction, purifying and the separation and leaching technology of active ingredient in a kind of Chinese medicine.

Background technology

Liquid-liquid technique is a most basic a kind of fractionation.The essence of this method is to add in the aqueous solution of separated material and the mutually not melt and dissolved organic solvent of water, makes that one or more components enter organic facies in the aqueous solution, and another kind of component is still stayed aqueous phase, thereby reaches the purpose of separation.Liquid-liquid technique is one of isolation technics that generally adopts in the chemical industry, in biochemical industry, genetic engineering, also there is it to use widely, yet, the stoste of most of biological products is low concentrations and has bioactive, need under low temperature or room temperature condition, carry out enrichment, separation, thereby conventional abstraction technique is restricted in these Application for Field.Based on this present situation, people have been developed double-aqueous phase system, and it is a kind ofly can keep bioactive a kind of novel liquid-liquid extraction isolation technics.But aqueous two phase extraction technique has also been run into some technical barriers in application and research process.For example: easily emulsification, the time that is separated are long, become the cost of gathering compound higher, and the most of viscosity of water soluble polymer are bigger, are difficult for quantitatively controlling etc., thereby are difficult to be applied in large-scale production and promote.Also seem awkward during the fat-soluble active ingredient of conventional extractive technique in extracting Chinese herbal medicine, because often contain moisture in the general Chinese herbal medicine, this makes fat-soluble organic matter be difficult to enter into cell interior, thereby can not be effectively and fully extract wherein active ingredient, be unfavorable for being applied in the large-scale production practices.

Summary of the invention

In order to remedy the deficiency of traditional extraction technology and emerging aqueous two phase extraction technique, we have invented a kind of organic matter-water-matter-water-organic matter purification extraction and separation technology.At present, this technology yet there are no relevant document and carried out report.Its basic principle is as follows: two kinds of visibly different organic matters of hydrophilic nmature and water mix by a certain percentage, can allow solution be divided into two-phase or multiphase system, according to object distribution coefficient difference therein, thereby can realize purifies and separates and the purpose of extracting target substance.According to experiment, this separation system is a kind of three-phase system basically, but becomes mutually very fast between first and second phase, can form basicly stable two phase liquid greatly within tens of seconds; Second and third becomes then comparatively slow mutually between mutually, becomes the phase time not wait from several minutes to a few hours.We utilize first and second to coordinate to come isolation of target substances with the characteristics that are difficult for emulsification fast mutually.In this sense, this system is equivalent to a double-aqueous phase system.

Investigate alcohol-water-chloroform, find that as long as three's ratio is suitable, solution system just can become phase when methanol-water-chloroform, acetone-water-chloroform and n-butanol-water-chloroform system.Aspect the proportionate relationship by changing three in this system of organic matter-water-organic matter and select different organic matters, the active ingredient of this method in extraction and purifies and separates Chinese medicine, boundless application prospect will be arranged.

The invention has the beneficial effects as follows, program is simple, realize easily, with low cost, separate rapidly and be difficult for emulsification, can be applied in the production practices of scale; Can reduce use, help preserving the ecological environment poisonous and harmful substance.

Applicating example: one, utilize this system of methanol-water-chloroform to extract active ingredient in Radix Glycyrrhizae and carry out thin-layer chromatography when differentiating, obtained very ideal effect." Chinese pharmacopoeia or " Chinese veterinary drug allusion quotation " are extracted the method that active ingredient is used in the Radix Glycyrrhizae: add a certain amount of ether and methyl alcohol respectively sample was added hot reflux each 1 hour, water dissolving behind the filtrate evaporate to dryness, adding n-butanol then extracts for several times, wash n-butanol liquid again with water for several times, also want the evaporate to dryness n-butanol at last.This method program is loaded down with trivial details, and is consuming time more, need use more harmful substance.During washing n-butanol liquid, if hold bad, washing excessively, can make water and n-butanol mutually emulsification mix can't layering.The n-butanol boiling point is higher, and rate of volatilization is slower, is difficult for evaporate to dryness.But, use methanol-water-chloroform system to carry out, will be simple many.Method is as follows: get an amount of Radix Glycyrrhizae, add 95% ethanol, ultrasonic processing 30 minutes filters, and residue washs merging filtrate 2-3 time with 95% ethanol.Evaporate to dryness, residue dissolves with methyl alcohol.Get supernatant, add chloroform, the vibration mixing adds an amount of water again, fully vibration back standing demix.Getting the upper strata differentiates just passable mutually.

Clearly, if in production practices, use a kind of method in back to extract active ingredient in the Radix Glycyrrhizae on a large scale, will save a large amount of time and cost.The scheme of our design is: add the ethanolic solution of proper proportion in Radix Glycyrrhizae, ultrasonic processing 30 minutes concentrates recovery ethanol through reducing pressure.Residue adds a certain amount of chloroform after dissolving with methyl alcohol, adds an amount of water behind the mixing again and makes solution be divided into two-phase up and down, gets upper strata phase evaporate to dryness and just can obtain crude extract, carries out fine finishining through suitable method again and gets final product.

Two, utilize methanol-water-chloroform separate in the Eupatorium adenophorum active ingredient also good effect.Method is as follows: get 20.05 gram samples (sample source Yu Guanling opens the purple eupatoria feed of rich Technology Park in the county), add methyl alcohol chloroform (1: 1) solution, ultrasonic 20 handle minute.Filter, residue washs with the small amount of methanol chloroform soln.Merging filtrate is put in the 250mL extraction flask, adds 10mL water, vibration, and standing demix is emitted lower floor's chloroform in a triangular flask, and the upper strata methanol-water is put in another triangular flask mutually.The washing of chloroform addition low amounts of water as emulsification, can add an amount of methyl alcohol, the vibration layering, and upper solution is integrated with in the methanol-water phase solution, is the methanol-water extract; Lower floor is the chloroform extract.Two kinds of solution are all used the water-bath evaporate to dryness, are respectively methanol-water extract and chloroform extract.Baking is 3.5 hours in 105 ℃ of drying boxes, weighs, and calculates content, is respectively 3.89% and 1.88% of institute's sample thief total amount.The chloroform extract that takes a morsel with the chloroform dissolving, adds the concentrated sulfuric acid, and upper strata chloroform soln virescence, this solution are put on silica gel g thin-layer plate, and it is red that fluorescence shows down; Lower floor is a concentrated sulfuric acid solution, and color is orange-yellow, adds 1% vanillic aldehyde concentrated sulfuric acid solution, adds methyl alcohol again, the solution becomes aubergine.Get chloroform extract 4 μ L, put on the polyamide lamellae, launch for solvent, take out, dry with chloroform methanol solution (24: 1).Inspect under the 365nm ultraviolet light, find three fluorescence spots, front end is the red fluorescence spot, and middle part and middle and lower part are the yellow-green fluorescence spot.Methanol-water is carried thing and is launched with method, launches to be with three fluorescence spots mainly also to occur, and middle front part is two yellow-green fluorescence spots, and the lower end is a blue-fluorescence spot, as seen, the polarity of these three kinds of materials a little less than.The spot of chromatographic band tail end shows expansion and incomplete, perhaps because polarity is too strong, also has some components not have separated opening.

The methanol-water extract adds the concentrated sulfuric acid and promptly produces white precipitate, the high temperature heating, white precipitate disappears, fling to obtain behind the first alcohol and water little liquid that shows flaxen slightly viscosity, continue heating and promptly produce a large amount of black smokes.Behind the methanol-water extract evaporate to dryness is faint yellow solid, and special fragrant (being similar to certain tea perfume (or spice)) is arranged, and does not have the distinctive pungent flavor of Eupatorium adenophorum.The aqueous solution adds ferric trichloride reagent and shows blackish green; Add the concentrated sulfuric acid and produce white precipitate.Make infrared spectrum with the KBr film-making, 3300,1560, stronger absorption is arranged near 1420,1272.9,1124.4, there is absworption peak at 1049.2 places.The chloroform extract has near 2850 two stronger absworption peaks, and other positions do not have tangible peak shape to occur.

The chloroform extract adds the concentrated sulfuric acid, and upper solution becomes green; Lower floor's solution becomes orange-yellow.Add 95% ethanolic solution, add water and make layering, the upper solution clarification is colourless, gets in right amount, and evaporate to dryness in the water-bath gets little liquid that shows flaxen slightly viscosity, and the high temperature heating promptly produces a large amount of black smokes.Lower floor's solution shows milky, must deceive the purple solid matter behind the evaporate to dryness in the water-bath.The floating thing of one deck adrift between the two-phase.

The specific embodiment

Character according to active ingredient in the Chinese medicine, select proper organic matter-water-organic-compound system, at first use wherein a kind of organic matter that the complex component that contains target component is extracted directly or indirectly, add then another kind not (easily) solubilized target composition but can effectively dissolve (with or precipitation) organic matter of other impurity compositions, add an amount of water at last, make solution be divided into immiscible two-phase, the solution that contains target component is extracted mutually, carry out suitable processing or refining getting final product.When extracting fat-soluble active ingredient, fat-soluble organic solvent and mixing by a certain percentage with its miscible hydrophilic organic solvent, extract target component in the middle of joining Chinese medicine, filter, filtrate adds water makes layering, extracts fat-soluble organic facies and processes and make with extra care and get final product.In concrete production practices, the addition sequence of solvent, mode and ratio cannot treat different things as the same, and mainly are to hold two principles: the one, can remove impurity to greatest extent and obtain object, and another is to make dicyandiamide solution can become phase.

Claims

1. organic matter-water-matter-water-organic matter purification, separation and leaching technology, its essential characteristics is: two kinds of visibly different organic matters of hydrophilic nmature and water are mixed by a certain percentage, make solution be divided into two-phase or multiphase system, according to object distribution coefficient difference therein, thereby realize purifies and separates and extract target substance.

2. purification, separation and leaching technology according to claim 1, it is characterized in that: solution system may be a kind of two-phase or multiphase system, mainly be to utilize first and second that form fast to come the separation and purification target substance mutually, in particular cases, also do not get rid of and use other to realize the purpose of separating mutually.

3. purification, separation and leaching technology according to claim 1 is characterized in that: two kinds of organic hydrophilic nmatures have notable difference, and can become phase after water mixes according to a certain percentage, no matter and its kind how.

4. purification, separation and leaching technology according to claim 1 is characterized in that: two kinds of organic hydrophilic nmatures have notable difference, and can become phase after water mixes according to a certain percentage, no matter and the mode of its mixing, addition sequence and ratio how.

5. purification, separation and leaching technology according to claim 1, it is characterized in that: two kinds of organic hydrophilic nmatures have notable difference, with can become phase after water mixes according to a certain percentage, target substance enters distribution coefficient difference after this system, no matter and the mode that it enters how.