CN103752039B - Self-supporting modularity chromatographic column and uses thereof - Google Patents
Self-supporting modularity chromatographic column and uses thereof Download PDFInfo
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- CN103752039B CN103752039B CN201410006266.7A CN201410006266A CN103752039B CN 103752039 B CN103752039 B CN 103752039B CN 201410006266 A CN201410006266 A CN 201410006266A CN 103752039 B CN103752039 B CN 103752039B
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Abstract
Present invention is disclosed a kind of self-supporting modularity chromatographic column and uses thereof.This modularity chromatographic column system forms by multiple plate comprising fixed phase stuffing is stacking, described plate is by adding binding agent and fluorescer in fixed phase stuffing, the mode solidification formings such as drying, sintering or polymerization, then the canonical shapes such as disk are polished into, form disk that is independent, self-support type, and then these disks are stacked up, form modularity chromatographic column.The program can observe the separation case of each composition of mixture in chromatographic column very easily, once reach required separation degree, just apart chromatographic column can be taken, by horizontal wash-out and collection, composition after quick acquisition is separated, thus shorten elution time, minimizing washing lotion consumption.
Description
Technical field
The present invention relates to some chemical composition to the naturally occurring animal of nature, plant, microorganism and metabolite thereof, and the various organic mixtures that the mankind are formed in all kinds of production and processing, scientific research process, carry out be separated or purify adopted method and instrument, particularly relate to chromatography and chromatograph, belong to organic mixture Separation & Purification technical field.
Background technology
Chromatography (Chromatography), also known as " chromatography ", " chromatography ", is a kind of high efficient separation technology for separating of multicomponent organic mixture.1906, Russia botanist Ci Weite (Mikhail Tsvet) has initiated this isolation technics, he pours one into the benzinum maceration extract of plant chloroplast pigment is equipped with in the thin straight glass column of calcium carbonate adsorbent, add pure benzinum again, let alone freely to flow down, original mixture starts the bands of a spectrum (i.e. the separation of phytochrome) being separated into different colours, and crosses pillar with friction speed, then carries out identification and analysis by bands of a spectrum Color pair mixture.At that time, Ci Weite called chromatogram this separating resulting, this separation method called after chromatography.Over more than 100 year, chromatographic theory is progressively set up and is developed, and chromatographic separation technology progressively realizes instrumentation, automation, high speed, and develops into analysis means from separation means, and range of application constantly expands.Although the object of chromatographic isolation is not limited to coloring matter already, " chromatography " this title is used always.
From principle angle, chromatography is a kind of method carrying out based on the uneven distribution of each component of mixture between two-phase (Stationary liquid and mobile phase) being separated.The prerequisite of uneven distribution is that each component is to the difference of two-phase affinity and the possibility to two-phase uneven distribution.Under the promotion of mobile phase, because the affinity of component each in mixture to two-phase is variant, they are just different through the flowing velocity (or the holdup time in Stationary liquid) of Stationary liquid, thus are separated.According to its separation principle, there are the methods such as adsorption chromatography, partition chromatography, ion-exchange chromatography.From application point, chromatographic technique is extensive application in research natural organic-compound.Some chemical composition in animal, plant, microorganism and metabolite thereof, it is the important sources of natural drug and natural additive for foodstuff, to their component structure and the research of physiologically active relation and the comprehensive utilization to natural materials, it is the research topic of current great potential, and to require the research and development of natural resources, to deepization, rapid, milligram ammonia future development, to serve the mankind to obtain the efficient natural constituent of safety non-toxic.Chromatographic technique just meets this requirement, and is widely applied in many-sides such as separating mixture, purified compound, authenticating compounds.Along with the integrated application of the technology such as chromatographic theory and electronics, optics, computer, new chromatographic technique is also constantly occurring and development, and chromatographic technique is also well applied in fine chemistry industry and polymeric material field.
At present, chromatography is all from chromatographic column one end sample introduction, adopts suitable solvent to carry out wash-out, the different material in mixture is eluted to the other end from chromatographic column one end, then collects successively.This method elution time is longer, and solvent load is large, needs fraction collector device costly during collection.Particularly for the mixture of the chemical monomer composition of rough segmentation more complicated, such as natural plant extracts, or the chemical mixture that essence point composition is less, said method seems very uneconomical.Therefore, need to improve existing chromatographic column, to obtaining better effect.
Summary of the invention
Object of the present invention is exactly to solve the above-mentioned problems in the prior art, a kind of self-supporting modularity chromatographic column is provided, and apply it carries out isolation andpurification concrete using method to organic mixture, once the ingredients of a mixture separates in the middle of each separation module of this chromatographic column, just can by horizontal wash-out and collection, composition after quick acquisition is separated, thus shorten elution time, minimizing washing lotion consumption.
The technology of the present invention solution is: a kind of self-supporting modularity chromatographic column, form by multiple plate comprising fixed phase stuffing is stacking, described plate is by adding binding agent solidification forming after super-dry, sintering or polymerization in fixed phase stuffing, can polishing further in necessary situation.
Preferably, in the middle of above-mentioned self-supporting modularity chromatographic column, in the occasion of sample introduction and wash-out of pressurizeing, such as: subcritical or supercritical fluid needs to promote solvent with pressure, is wrapped in the chromatographic column outside built up by plate with the inert coating of chemically-resistant solvent.Like this, when pressurizeing sample introduction and wash-out, can pressurize in chromatographic column outside, this pressure, higher than sample introduction and wash-out pressure, ensures that solvent is flushed to the other end from one end, and can not from chromatographic column outside seepage.
More preferably, in the middle of above-mentioned self-supporting modularity chromatographic column, described binding agent is water formulation or organic solvent preparation, adds this water formulation or organic solvent preparation, be shaped after drying in fixed phase stuffing; Or described binding agent is solid pharmaceutical preparation, in fixed phase stuffing, add this solid pharmaceutical preparation, through sintering postforming; Or described binding agent is monomer formulation, in fixed phase stuffing, add this monomer formulation, by heating, UV light-induced or chemical action and being shaped after producing polymerisation.Specifically, described preparation includes but not limited to it is calcium sulfate, waterglass, glass dust, polyethylene or polyacrylic acid.
Again preferably, above-mentioned self-supporting modularity chromatographic column, the shape of described plate is disk; Further, first can be stacked by 1 or several plate and form 3 ~ 30 single modules, and then be stacked gradually by each module and form chromatographic column; And for ease of observing, described plate also added fluorescer before solidification forming.
Any one self-supporting modularity chromatographic column above, can be used for being separated organic mixture: first, pending mixture is dissolved in suitable solvent, then in the mode of dry method, that is: needing the mixture be separated to be dissolved in suitable solvent, the solution formed adds to a disk, then allows solvent evaporate into bone dry, from one end sample introduction of this modularity chromatographic column; Then, using the positive contrary with Stationary liquid absorption property or anti-phase solvent, subcritical or supercritical fluid as washing lotion, wash-out is carried out from the sample introduction end of chromatographic column.When adopting subcritical or supercritical fluid as washing lotion, need external pressure to promote solvent, now, can be wrapped in the inert coating of chemically-resistant solvent the chromatographic column outside built up by plate, to ensure that sample introduction and wash-out carry out smoothly.When mobile phase is eluted to the other end from sample introduction one end, stops wash-out, from each connecting portion, this modularity chromatographic column is disassembled, more individually rinse single module or each plate with solvent, after dry, obtain the individual components after being separated.
In above-mentioned application process, after the first round, wash-out completed, indivedual module can be assembled in new modularity chromatographic column again, change the washing lotion wash-out again that another kind is more suitable, separative efficiency is higher.This process can repeatedly (be equivalent to " multi-dimensional chromatograph "), until separating effect up to standard after, more individually rinse separation module with solvent, after dry, obtain required component.And, can cosolvent be added in the middle of the washing lotion that wash-out is used, to improve elution efficiency.
The present invention breaks through the completely concept of prior art about chromatography Stationary liquid used, be processed into the disk of stand alone type, self-supporting one by one, and then by these disk stack assembly, form a kind of self-supporting disc type modularity chromatographic column, and for the component separation to organic mixture.Once the ingredients of a mixture in the middle of each separation module of this modularity chromatographic column separately, just chromatographic column can be split and come, then wash-out and collection be carried out to each separation module, thus the composition after separation fast required for acquisition.
Compared with the chromatography of routine, the present invention operates simple and easy, and greatly can save the consumption of solvent or washing lotion, wash-out is more simple and efficient, and, do not need expensive fraction collector device.When only needing to be separated minority component in the middle of acquisition mixture, as long as carry out wash-out to a few separation module at these component places, solvent load is few, and energy is dry within a short period of time, and then obtains target component fast.
The present invention had both been applicable to the organic mixture of the comparison of ingredients complexity such as rough segmentation natural plant extracts, the organic mixture that also can smartly divide composition less, also be applicable to the organic mixture that separate targets comparison of ingredients is clear and definite, for the extraction of natural goods, the separation of known mixture, or the research and production etc. of unknown mixture is all a kind of well technical solution.
Detailed description of the invention
The present invention is directed to existing chromatographic column to be all a complete pillar, in rapid not, the sub-economic defect of some use occasions, to propose a kind of self-supporting modularity chromatographic column.Adopt this modularity chromatographic column, the separation case of each composition of mixture in chromatographic column can be observed very easily, once reach required separation degree, just apart chromatographic column can be taken, by horizontal wash-out and collection, composition after quick acquisition is separated, thus shorten elution time, minimizing washing lotion consumption.
According to technical solution of the present invention, (both can be first positive, as silica gel at selected Stationary liquid; Also can be anti-phase, as carbon 18) innerly add binding agent, and, selectively add fluorescer, solidification forming after super-dry, sintering or polymerization, after polishing, form plate that is independent, self-support type, then these plates are stacked up, form modularity chromatographic column.Time stacking, also first can be stacked by 1 or several plate and form single module, and then be stacked gradually by each single module and form chromatographic column.
Different according to the kind of binding agent, the preparation method of plate has multiple.When binding agent be water formulation or organic solvent preparation time, add this water formulation or organic solvent preparation in fixed phase stuffing, be shaped after drying, then polish further; If binding agent is solid pharmaceutical preparation, as: calcium sulfate, waterglass, glass dust, polyethylene or polyacrylic acid etc., then add this solid pharmaceutical preparation in fixed phase stuffing, through sintering postforming; Binding agent can also be monomer formulation, adds this monomer formulation in fixed phase stuffing, by heating, UV light-induced or chemical action, makes it to produce polymerisation and then solidified forming.
Below the several concrete preparation example of plate:
(1) being applicable to and appropriate Stationary liquid, add and be applicable to obtaining suspension with appropriate glass dust and liquid (such as water or ethanol), suspension is thoroughly stirred, liquid is allowed slowly to evaporate, then this mixture is put into sintering in baking oven (750 DEG C), become plate after sintering, plate can wear into suitable shape (such as disk);
(2) being applicable to and appropriate Stationary liquid, add and be applicable to and appropriate fluorescer, add again and be applicable to and appropriate glass dust and liquid (such as water or ethanol), obtain suspension, suspension is thoroughly stirred, allows liquid slowly evaporate, then this mixture is put into sintering in baking oven (750 DEG C), become the plate having fluorescer after sintering, plate can wear into suitable shape (such as disk);
(3) being applicable to and appropriate Stationary liquid, add and be applicable to and appropriate adhesive (such as polyacrylic acid, waterglass) and liquid (such as water or ethanol), obtain suspension, suspension is thoroughly stirred, allow liquid slowly and evaporate to dryness up hill and dale, become plate after solidification, plate can wear into suitable shape (such as disk);
(4) being applicable to and appropriate Stationary liquid, add and be applicable to and appropriate fluorescer, add again and be applicable to and appropriate adhesive (such as polyacrylic acid, waterglass) and liquid (such as water or ethanol), suspension, suspension is thoroughly stirred, then allows liquid slowly and evaporate to dryness up hill and dale, become the plate having fluorescer after solidification, plate can wear into suitable shape (such as disk).
According to technical solution of the present invention, in the middle of self-supporting modularity chromatographic column, the usage quantity of plate is unrestricted, and its concrete consumption should be determined according to use occasion and separation requirement, 3 ~ 5 pieces at least, 20 ~ 30 pieces at most; And the format diameter of each plate, thickness height, both can be the same or different, as long as can be convenient to assembling.The modularity chromatographic column that assembling is formed, is applicable to: the mixture of the chemical monomer composition of rough segmentation more complicated, such as natural plant extracts, or the organic mixture that essence point composition is less, or for separating of the clearer and more definite organic mixture of target component.
Its basic process is: first pending mixture is dissolved in suitable solvent, then from one end sample introduction of this modularity chromatographic column; Then, using the positive contrary with Stationary liquid absorption property or anti-phase solvent, subcritical or supercritical fluid as washing lotion, from the sample introduction end wash-out of chromatographic column, just wash-out can be stopped when mobile phase is eluted to the other end from sample introduction one end, then from each connecting portion, this modularity chromatographic column is disassembled, individually rinse each separation module with solvent, after dry, obtain the individual product after being separated.In addition, the mode of " crossing post dry method loading " can also be adopted to carry out sample introduction and wash-out: be first mix sample, pending mixture is dissolved in suitable solvent and forms solution, then this solution is added to above a preformed disk, allow solvent evaporate into bone dry again, make sample adsorption on this disk; Then, using this block disk with sample as the disk of self-supporting disc type modularity chromatographic column liquid feeding end, then by washing lotion, wash-out separation is carried out to it.In elution process, in the middle of washing lotion, cosolvent can also be added.
According to the concrete condition of used mobile phase, system elutions can utilize the modes such as the attraction (Capillary attraction) of Action of Gravity Field (Gravity), capillary, promotes mobile phase and carry out wash-out under pressurization or non-pressurized situation.Specifically, when utilizing gravity mode, after sample introduction, mobile phase is wash-out from the top to the bottom; When utilizing the attraction of capillary, after sample introduction, mobile phase is from bottom to top wash-out.Under pressurized condition, the chromatographic column outside built up by disk is wrapped in the inert coating of chemically-resistant solvent, expose two ends, then radial pressurization (radial compression to add pressure), then utilize pressure that mobile phase is eluted to the other end from one end of sample introduction, be equivalent to superpressure thin-layer chromatography (Overpressure thin layer chromatography) principle.And elution process is applicable multidimensional liquid phase method also, after completing each module of first round wash-out and separating, indivedual module or disk can be installed on a new row combination module, change the mobile phase wash-out again that another kind is more suitable, separative efficiency is higher.Said process can repeat several times, until need the product be separated to reach desirable purity.This repetitive process is more close to the ideal concepts of moving-bed chromatographic.
After wash-out completes, successively or repeatedly can be combined detection separative efficiency.Time in the middle of Stationary liquid not containing fluorescer, FID scanning, IR scanning, scanning of the mass spectrum can be used, or detect again after wash-out; Time in the middle of Stationary liquid containing fluorescer, as long as by ultraviolet light irradiation chromatographic column, under suitable wavelength condition, just can observe the separation degree of mixture, see which separation module active (UV active) compound of ultraviolet is positioned in.
If separation degree is desirable, under can using solvent that active for ultraviolet (UV active) compound is rinsed from separation module immediately; If separation degree is undesirable, with more separation modules, chromatographic column can be extended, continue wash-out, until separation degree is desirable.Or, the separation module having ultraviolet to enliven compound is installed in a new row modularity chromatographic column, changes another kind of more suitable, that separative efficiency is higher mobile phase and continue wash-out, until reach desirable separating effect.Certainly, if target components is non-ultraviolet enliven compound, implement with reference to above method.
In sum, present invention is disclosed a kind of self-supporting modularity chromatographic column, especially a kind of self-supporting disc type modularity chromatographic column, and the concrete using method applying that it carries out isolation andpurification to organic mixture.Utilize technical solution of the present invention, once the ingredients of a mixture separates in the middle of each separation module of this modularity chromatographic column, just chromatographic column can be split and come, then wash-out and collection be carried out to each separation module, thus the composition after separation required for obtaining fast.
Compared with the chromatography of routine, the present invention operates simple and easy, greatly can save the consumption of solvent or washing lotion, and wash-out is more quick, and, do not need expensive fraction collector device.When only needing to be separated minority component in the middle of acquisition mixture, as long as carry out wash-out to a few separation module at these component places, solvent load is few, and energy is dry within a short period of time, and then obtains target component fast.
The present invention had both been applicable to the organic mixture of the comparison of ingredients complexity such as rough segmentation natural plant extracts, the organic mixture that also can smartly divide composition less, also be applicable to the organic mixture that separate targets comparison of ingredients is clear and definite, Nature inorganic bone, known mixture are separated, or the research and production etc. of unknown mixture is all a kind of well technical solution.Certainly, the above is only the prominent example of application technical solution of the present invention, allly takes equivalent replacement or equivalent transformation and the technical scheme that formed, all drops within the scope of protection of present invention.
Claims (11)
1. a self-supporting modularity chromatographic column, it is characterized in that: this modularity chromatographic column system forms by multiple plate comprising fixed phase stuffing is stacking, described plate is by adding binding agent solidification forming after super-dry, sintering or polymerization in fixed phase stuffing.
2. self-supporting modularity chromatographic column according to claim 1, is characterized in that: in the occasion of sample introduction and wash-out of pressurizeing, be wrapped in the chromatographic column outside built up by plate with the inert coating of chemically-resistant solvent.
3. self-supporting modularity chromatographic column according to claim 1 and 2, is characterized in that: described binding agent is water formulation, organic solvent preparation, solid pharmaceutical preparation or monomer formulation.
4. self-supporting modularity chromatographic column according to claim 3, is characterized in that: described preparation is calcium sulfate, waterglass, glass dust, polyethylene or polyacrylic acid.
5. self-supporting modularity chromatographic column according to claim 1 and 2, is characterized in that: the shape of described plate is disk.
6. self-supporting modularity chromatographic column according to claim 1 and 2, is characterized in that: first stacked by 1 or several plate and form single module, and then is stacked gradually by each module and form chromatographic column.
7. self-supporting modularity chromatographic column according to claim 6, is characterized in that: the quantity of described single module is 3 ~ 30.
8. self-supporting modularity chromatographic column according to claim 1 and 2, is characterized in that: described plate also added fluorescer before solidification forming.
9. the purposes of any one self-supporting modularity chromatographic column described in claim 1 ~ 8, it is characterized in that: for separating of organic mixture, first, pending mixture is dissolved in suitable solvent, then from one end sample introduction of this modularity chromatographic column, then, with the positive contrary with Stationary liquid absorption property or anti-phase solvent, subcritical or supercritical fluid is as washing lotion, wash-out is carried out from the sample introduction end of chromatographic column, when mobile phase is eluted to the other end from sample introduction one end, stop wash-out, from each connecting portion, this modularity chromatographic column is disassembled, single module or each plate is individually rinsed again with solvent, the individual components after being separated is obtained after dry.
10. the purposes of self-supporting modularity chromatographic column according to claim 9, it is characterized in that: after first round wash-out completes, indivedual module is assembled in new modularity chromatographic column again, changes the washing lotion wash-out again that another kind is more suitable, separative efficiency is higher; This process can repeatedly, until separating effect up to standard after, more individually rinse single module or each plate with solvent, after dry, obtain required component.
The purposes of 11. self-supporting modularity chromatographic columns according to claim 9, is characterized in that: in described elution process, be added with cosolvent in the middle of washing lotion.
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| CN202490463U (en) * | 2012-03-26 | 2012-10-17 | 常州三泰科技有限公司 | Liquid chromatographic separation column capable of being manually assembled |
| CN203043621U (en) * | 2013-02-04 | 2013-07-10 | 北京创新通恒科技有限公司 | Liquid chromatographic column |
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| JPS63285465A (en) * | 1987-05-04 | 1988-11-22 | ケマップ アクチエンゲゼルシャフト | Adsorption column for chromatographic separation and working method thereof |
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