CN101614711A - A kind of automatic three-stage chromatograph - Google Patents
A kind of automatic three-stage chromatograph Download PDFInfo
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Abstract
A kind of automatic three-stage chromatograph comprises: the first order is separated and is comprised that separate pump 1, sampling pump, valve-1, one-level separating column, flow point transfer valve 1 and six one-level sample component receive post; The second level is separated and is comprised separate pump 2, valve-2, flow point transfer valve 2 and secondary separating column; The third level separates and comprises separate pump 3-1, valve-3-1 and separate pump 3-2, valve-3-2; Flow point transfer valve 3; Six three grades of separating columns; Two part flow arrangements; Two pick-up units and two automatic collecting devices; Above-mentioned steps is finished by microcomputer PC control.Automatic three-stage chromatograph of the present invention can be used for purifying complex organic synthesis compound, and drug candidate matter such as native compound and biofermentation product are to reach the separation and purification of monomeric compound.
Description
Technical field
The present invention relates to a kind of automatic three-stage chromatograph, this automatic three-stage chromatograph can be used for purifying complex organic synthesis compound, and drug candidate matter such as native compound and biofermentation product are to reach the separation and purification of monomeric compound.
Background technology
The chromatography chromatogram purification comprises chromatography chromatographic equipment and two aspects of chromatography chromatographic media.The clastotype of chromatography chromatogram is by the decision of the chromatographic media that is used, and single-stage chromatography chromatogram is used single separating medium, for example in the single-stage reverse-phase chromatography, can be to use a kind of based on the normal octane medium as the anti-phase chromatographic media of single-stage.
The operation of the traditional chromatography purifier apparatus of one cover is the single-stage single channel, promptly interior at one time just sample of chromatographic purifying.So purifying speed is slow, separation efficiency is low.Traditional multistage chromatographic resolution is the combination of the chromatography chromatogram of several model identical, a kind of chromatographic resolution pattern and another chromatographic resolution pattern usually can not be compatible fully, normal hexane solvent system that for example uses in the normal-phase chromatography and the methanol-water system in the reverse-phase chromatography are inconsistent, make that the chromatography chromatogram of different mode can not continued operation, and must independently carry out in the tomographic system respectively.
In the R﹠D process of medicine, a wherein crucial step is that compound separation to be selected and purifying are obtained the pure product compound of monomer.A lot of materials to be selected, the crude extract of natural products for example, composition are very complicated usually, particularly the extraction of some microcomponents with separate difficult unusually especially.Only rely on one-level chromatography chromatographic resolution to be difficult to obtain the pure product of monomer.Usually need multistage chromatography chromatographic resolution.
In recent years in the chromatography chromatographic field, the means that improve separation efficiency may be summarized to be two classes 1) use novel chromatography media, the disengaging time 2 of shortening single-stage chromatography) flux of raising single-stage chromatography, for example single-stage multicolumn parallel chromatography instrument and single-stage multicolumn series connection suitching type chromatograph.But these chromatographs still are to use single-stage chromatography chromatographic separation technology.
The objective of the invention is to provide a kind of automatic three-stage chromatograph, realize the combination of several different chromatographic resolution patterns, i.e. three grades of different chromatography separation medias continuous separation in same chromatograph, the monomer purifying of compatibility, improvement chromatography flux and the separation efficiency between the improvement chromatography dielectric solvent system, the medicine sample to be selected of improvement complexity.
Summary of the invention
The objective of the invention is to provide a kind of automatic three-stage chromatograph, realize the combination of several different chromatographic resolution patterns, i.e. three grades of different chromatography separation medias continuous separation in same chromatograph, the monomer purifying of compatibility, improvement chromatography flux and the separation efficiency between the improvement chromatography dielectric solvent system, the medicine sample to be selected of improvement complexity.The separation and purification process that this instrument carries out is a continuously dynamic separation and purification process.
Automatic three-stage chromatograph of the present invention comprises:
(1) first order is separated and is comprised that separate pump 1, sampling pump, valve-1, one-level separating column, flow point transfer valve 1 and six one-level sample component receive post;
(2) second level is separated and is comprised separate pump 2, valve-2, flow point transfer valve 2 and secondary separating column;
(3) third level separates and comprises separate pump 3-1, valve-3-1 and separate pump 3-2, valve-3-2; Flow point transfer valve 3; Six three grades of separating columns; Two part flow arrangements; Two pick-up units and two automatic collecting devices;
(4) above-mentioned steps is finished by microcomputer (PC) control.
As further improvement in the technical proposal, the described third level separates and comprises separate pump 3-1, valve-3-1 and separate pump 3-2, valve-3-2; Flow point transfer valve 3; Six three grades of separating columns are called three grades of separating column 3-1, three grades of separating column 3-2, three grades of separating column 3-3, three grades of separating column 3-4, three grades of separating column 3-5 and three grades of separating column 3-6; Two part flow arrangements are called part flow arrangement 3-1, part flow arrangement 3-2; Two pick-up units are called pick-up unit 3-1, pick-up unit 3-2; Two automatic collecting devices are called automatic collecting device 3-1, automatic collecting device 3-2.
As further improvement in the technical proposal, the described third level separates and comprises separate pump 3-1, valve-3-1 and separate pump 3-2, valve-3-2; Flow point transfer valve 3; Six three grades of separating columns are called three grades of separating column 3-1, three grades of separating column 3-2, three grades of separating column 3-3, three grades of separating column 3-4, three grades of separating column 3-5 and three grades of separating column 3-6; These six three grades of separating columns are connected to two part flow arrangements, part flow arrangement 3-1 and part flow arrangement 3-2, wherein three grades of separating column 3-1, three grades of separating column 3-2, three grades of separating column 3-3 are connected with part flow arrangement 3-1, and three grades of separating column 3-4, three grades of separating column 3-5, three grades of separating column 3-6 are connected with part flow arrangement 3-2; Part flow arrangement is connected with pick-up unit, and part flow arrangement 3-1 is connected with pick-up unit 3-1, and part flow arrangement 3-2 is connected with pick-up unit 3-2; Pick-up unit is connected with automatic collecting device, and pick-up unit 3-1 is connected with automatic collecting device 3-1, and pick-up unit 3-2 is connected with automatic collecting device 3-2.
As further improvement in the technical proposal, the described first order is separated, and it is characterized in that using an one-level separating column.
As further improvement in the technical proposal, described component of separating through the first order through flow point transfer valve 1, is enriched in six one-level sample component and receives in the post, its described flow point transfer valve 1, the volume that makes every one-level sample component receive enriched composition in the post through microcomputer PC control is equal to.
As further improvement in the technical proposal, described six one-level sample component receive post, be called sample 1, sample 2, sample 3, sample 4, sample 5 and sample 6, the filler that these six one-level sample component receive in the post is a chromatographic media, this chromatographic media reaches maximization and enrichment diffusion minimum to the enrichment of component, usually this chromatographic media is the filler that material to be separated is had strong affinity, straight chain saturated alkane class medium for example, described straight chain saturated alkane class medium comprises: octadecyl silane, the octane bonded silica gel, the phenethyl bonded silica gel, pentafluorophenyl group, hexane, butane, dodecyl.
As further improvement in the technical proposal, the described component that is enriched in the one-level sample component reception post enters the second level respectively and separates, the chromatographic resolution pattern that separate the second level is different with the chromatographic resolution pattern that the first order is separated, usually the chromatographic resolution pattern of first order separation is by ion-exchange or ion exclusion, chargeding performance according to composition separates, or separates by the molecular weight size by size exclusion chromatograph; The second level separate the chromatographic resolution pattern typically use unique hydrophilic filler, utilize hydrophilic Interaction Chromatography mechanism (HILIC:HydrophILic Interaction Chromatography or HydrophilicInteraction LIquid Chromatography) to separate.Described hydrophilic filler comprises: contain the styrene-divinylbenzene copolymer of benzene sulfonic acid group, the styrene-divinylbenzene copolymer that contains the quaternary amine group, amide-containing styrene-divinylbenzene copolymer, contain amino styrene-divinylbenzene copolymer, the bonded silica gel that contains the benzene sulfonic acid group, the bonded silica gel that contains the quaternary amine group, amide-containing bonded silica gel, contain amino bonded silica gel.
As further improvement in the technical proposal, described component of separating through the second level directly enters six three grades of separating columns and carries out the third level and separate, and the filler in described six three grades of separating columns is that particle diameter is the chromatographic media of 5 μ m～15 μ m.
As further improvement in the technical proposal, it is a quick detachment process that the described third level separates, and separation cycle is generally 3～5 minutes.
The chromatographic resolution pattern that the third level separates adopts efficient reverse-phase chromatography to realize.This grade separates the high-efficiency column that uses 5～10cm column length, separating medium is a straight chain saturated alkane class filler, and described straight chain saturated alkane class filler comprises: octadecyl silane, octane bonded silica gel, phenethyl bonded silica gel, pentafluorophenyl group, hexane, butane, dodecyl.
As further improvement in the technical proposal, receive by automatic collecting device 3-1 and automatic collecting device 3-2 respectively after described component of separating through the third level detects by pick-up unit 3-1 and pick-up unit 3-2, the collection form that its described automatic collecting device adopted can for receive entirely, time band or receive according to the appearance automatic switchover position at peak.
Description of drawings
Further specify below in conjunction with drawings and Examples.
Fig. 1 is an automatic three-stage chromatograph synoptic diagram of the present invention.
Separate pump 1, sampling pump, valve-1, one-level separating column, flow point transfer valve 1 and six one-level sample component receive post, are called sample 1, sample 2, sample 3, sample 4, sample 5 and sample 6;
Separate pump 2, valve-2, flow point transfer valve 2 and secondary separating column;
Separate pump 3-1, valve-3-1 and separate pump 3-2, valve-3-2; Flow point transfer valve 3; Six three grades of separating columns are called three grades of separating column 3-1, three grades of separating column 3-2, three grades of separating column 3-3, three grades of separating column 3-4, three grades of separating column 3-5 and three grades of separating column 3-6; Two part flow arrangements are called part flow arrangement 3-1, part flow arrangement 3-2; Two pick-up units are called pick-up unit 3-1, pick-up unit 3-2; Two automatic collecting devices are called automatic collecting device 3-1, automatic collecting device 3-2;
The said equipment is controlled by microcomputer (PC);
Moving phase 1, moving phase 2, moving phase 3, sample.
Embodiment
Automatic three-stage chromatograph, at first moving phase 1 enters the one-level separating column by separate pump 1, and sample enters the one-level separating column by sampling pump and valve-1 simultaneously, carries out the first order and separates; Treat sample after the one-level separating column separates, the effluent component is stored in six one-level sample component respectively by flow point transfer valve 1 and receives in the post, is called sample 1, sample 2, sample 3, sample 4, sample 5 and sample 6.Be stored in the component in the one-level sample component reception post,, enter the secondary separating column simultaneously, carry out the second level and separate with the moving phase 2 that enters by separate pump 2 and valve-2 by flow point transfer valve 2.Because the chromatographic resolution pattern that separate the second level is different from the chromatographic resolution pattern that the first order is separated, therefore in separating, the first order do not obtain the material of monomer separation, will further obtain separating.Treat that sample is after the secondary separating column separates, the effluent component is by flow point transfer valve 3, directly enter three grades of separating columns, carrying out the third level separates: the component that separates through the second level enters three grades of separating column 3-1, three grades of separating column 3-2, three grades of separating column 3-3 respectively simultaneously through flow point transfer valve 3 and moving phase 3 by separate pump 3-1, valve-3-1, carries out the third level and separates; The component that separates through the second level enters three grades of separating column 3-4, three grades of separating column 3-5 and three grades of separating column 3-6 respectively simultaneously through flow point transfer valve 3 and moving phase 3 by separate pump 3-2, valve-3-2, carries out the third level and separates.
The component of separating respectively through three grades of separating column 3-1, three grades of separating column 3-2, three grades of separating column 3-3 enters part flow arrangement 3-1 in succession, and detects by pick-up unit 3-1 one by one, and the component after is collected through automatic collecting device 3-1 after testing; The component of separating respectively through three grades of separating column 3-4, three grades of separating column 3-5, three grades of separating column 3-6 enters part flow arrangement 3-2 in succession, and detects by pick-up unit 3-2 one by one, and the component after is collected through automatic collecting device 3-2 after testing.
By control flow point transfer valve 3, the component of separating through the second level enters three grades of separating columns respectively, carries out the third level and separates.It comprises: i-is by control flow point transfer valve 3, and the component of separating through the second level enters three grades of separating column 3-1 by flow point transfer valve 3, and moving phase 3 enters separating column 3-1 by separate pump 3-1 and valve-3-1 simultaneously, separates fast; Ii-is by control flow point transfer valve 3, and the component of separating through the second level enters three grades of separating column 3-2 by flow point transfer valve 3, and moving phase 3 enters separating column 3-2 by separate pump 3-1 and valve-3-1 simultaneously, separates fast; Iii-is by control flow point transfer valve 3, and the component of separating through the second level enters three grades of separating column 3-3 by flow point transfer valve 3, and moving phase 3 enters separating column 3-3 by separate pump 3-1 and valve-3-1 simultaneously, separates fast; Iv-is by control flow point transfer valve 3, and the component of separating through the second level enters three grades of separating column 3-4 by flow point transfer valve 3, and moving phase 3 enters separating column 3-4 by separate pump 3-2 and valve-3-2 simultaneously, separates fast; V-is by control flow point transfer valve 3, and the component of separating through the second level enters three grades of separating column 3-5 by flow point transfer valve 3, and moving phase 3 enters separating column 3-5 by separate pump 3-2 and valve-3-2 simultaneously, separates fast; Vi-is by control flow point transfer valve 3, and the component of separating through the second level enters three grades of separating column 3-6 by flow point transfer valve 3, and moving phase 3 enters separating column 3-6 by separate pump 3-2 and valve-3-2 simultaneously, separates fast.Above-mentioned six steps are continuous Dynamic Separation process.
The filler that described six one-level sample component receive in the post is a straight chain saturated alkane class medium, and described straight chain saturated alkane class filler comprises: octadecyl silane, octane bonded silica gel, phenethyl bonded silica gel, pentafluorophenyl group, hexane, butane, dodecyl.
The chromatographic resolution pattern that the described first order is separated is by ion-exchange or ion exclusion, separates according to the chargeding performance of composition, or separates by the molecular weight size by size exclusion chromatograph; The chromatographic resolution pattern that separate the second level typically uses unique hydrophilic filler, utilizes hydrophilic Interaction Chromatography mechanism (HILIC:HydrophILic Interaction Chromatography or Hydrophilic InteractionLIquid Chromatography) to separate.Described hydrophilic filler comprises: contain the styrene-divinylbenzene copolymer of benzene sulfonic acid group, the styrene-divinylbenzene copolymer that contains the quaternary amine group, amide-containing styrene-divinylbenzene copolymer, contain amino styrene-divinylbenzene copolymer, the bonded silica gel that contains the benzene sulfonic acid group, the bonded silica gel that contains the quaternary amine group, amide-containing bonded silica gel, contain amino bonded silica gel.
Filler in described six three grades of separating columns is that particle diameter is the chromatographic media of 5～15 μ m.
It is a quick detachment process that the described third level separates, and separation cycle is generally 3～5 minutes.The third level separates the high-efficiency column that uses 5～10cm column length, separating medium is a straight chain saturated alkane class filler, and described straight chain saturated alkane class filler comprises: octadecyl silane, octane bonded silica gel, phenethyl bonded silica gel, pentafluorophenyl group, hexane, butane, dodecyl.
The collection line that described automatic collecting device adopted can for complete receive, time band or receive according to the appearance automatic switchover position at peak.
Obviously, the above embodiment of the present invention only is for example of the present invention clearly is described, and is not to be qualification to embodiments of the present invention.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here can't give exhaustive to all embodiments.Everyly belong to the row that conspicuous variation that technical scheme of the present invention extends out or change still are in protection scope of the present invention.
Claims (8)
1, a kind of automatic three-stage chromatograph is characterized in that, comprises as the lower part:
(1) first order is separated and is comprised that separate pump 1, sampling pump, valve-1, one-level separating column, flow point transfer valve 1 and six one-level sample component receive post;
(2) second level is separated and is comprised separate pump 2, valve-2, flow point transfer valve 2 and secondary separating column;
(3) third level separates and comprises separate pump 3-1, valve-3-1 and separate pump 3-2, valve-3-2,3, six three grades of separating columns of flow point transfer valve, two part flow arrangements, two pick-up units and two automatic collecting devices;
(4) above-mentioned steps is finished by microcomputer PC control.
2,, it is characterized in that third level separation comprises separate pump 3-1, valve-3-1 and separate pump 3-2, valve-3-2 according to the described automatic three-stage chromatograph of claim 1; Flow point transfer valve 3; Six three grades of separating columns are called three grades of separating column 3-1, three grades of separating column 3-2, three grades of separating column 3-3, three grades of separating column 3-4, three grades of separating column 3-5 and three grades of separating column 3-6; These six three grades of separating columns are connected to two part flow arrangements, part flow arrangement 3-1 and part flow arrangement 3-2, wherein three grades of separating column 3-1, three grades of separating column 3-2, three grades of separating column 3-3 are connected with part flow arrangement 3-1, and three grades of separating column 3-4, three grades of separating column 3-5, three grades of separating column 3-6 are connected with part flow arrangement 3-2; Part flow arrangement is connected with pick-up unit, and part flow arrangement 3-1 is connected with pick-up unit 3-1, and part flow arrangement 3-2 is connected with pick-up unit 3-2; Pick-up unit is connected with automatic collecting device, and pick-up unit 3-1 is connected with automatic collecting device 3-1, and pick-up unit 3-2 is connected with automatic collecting device 3-2.
3,, it is characterized in that it is an one-level separating column that the first order is separated employed according to the described automatic three-stage chromatograph of claim 1.
4, according to the described automatic three-stage chromatograph of claim 1, it is characterized in that six one-level sample component receive post, be called sample 1, sample 2, sample 3, sample 4, sample 5 and sample 6, the filler that these six one-level sample component receive in the post is a straight chain saturated alkane class medium, and described straight chain saturated alkane class medium comprises: octadecyl silane, octane bonded silica gel, phenethyl bonded silica gel, pentafluorophenyl group, hexane, butane, dodecyl.
5, according to the described automatic three-stage chromatograph of claim 1, it is characterized in that the chromatographic resolution pattern that the chromatographic resolution pattern of separating the second level and the first order separate is different, the chromatographic resolution pattern that the first order is separated is by ion-exchange or ion exclusion, chargeding performance according to composition separates, or separates by the molecular weight size by size exclusion chromatograph; The second level separate the chromatographic resolution pattern be to use unique hydrophilic filler, utilize hydrophilic Interaction Chromatography mechanism to separate.Described hydrophilic filler comprises: contain the styrene-divinylbenzene copolymer of benzene sulfonic acid group, the styrene-divinylbenzene copolymer that contains the quaternary amine group, amide-containing styrene-divinylbenzene copolymer, contain amino styrene-divinylbenzene copolymer, the bonded silica gel that contains the benzene sulfonic acid group, the bonded silica gel that contains the quaternary amine group, amide-containing bonded silica gel, contain amino bonded silica gel.
6, according to the described automatic three-stage chromatograph of claim 1, it is characterized in that the third level separates, the filler in six three grades of separating columns is that particle diameter is the chromatographic media of 5 μ m～15 μ m.
7, according to the described automatic three-stage chromatograph of claim 1, it is characterized in that it is a quick detachment process that the third level separates, separation cycle is generally 3～5 minutes.The chromatographic resolution pattern that the third level separates adopts efficient reverse-phase chromatography to realize.This grade separates the high-efficiency column that uses 5～10cm column length, and separating medium is a straight chain saturated alkane class filler.Described straight chain saturated alkane class filler comprises: octadecyl silane, octane bonded silica gel, phenethyl bonded silica gel, pentafluorophenyl group, hexane, butane, dodecyl.
8, according to the described automatic three-stage chromatograph of claim 1, it is characterized in that collection line that automatic collecting device 3-1 and automatic collecting device 3-2 adopt can for receive entirely, time band or receive according to the appearance automatic switchover position at peak.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102008837A (en) * | 2010-09-30 | 2011-04-13 | 中国科学院昆明植物研究所 | Separating preparative chromatography apparatus comprising serial preparative chromatography column |
| CN102172516A (en) * | 2011-03-28 | 2011-09-07 | 天津博纳艾杰尔科技有限公司 | Novel resin-coated silica-gel adsorption material |
| CN102441294A (en) * | 2010-09-30 | 2012-05-09 | 中国科学院昆明植物研究所 | Separation and preparation chromatograph containing series static axial compression (SAC) preparative chromatographic columns |
| CN102441295A (en) * | 2010-09-30 | 2012-05-09 | 中国科学院昆明植物研究所 | Separation and preparation chromatograph containing series dynamic axial compression (DAC) preparative chromatographic columns |
| CN104407083A (en) * | 2014-11-28 | 2015-03-11 | 天津博纳艾杰尔科技有限公司 | Full-automatic separation system and application of full-automatic separation system in separation of polar components in edible oil |
| WO2017028357A1 (en) * | 2015-08-18 | 2017-02-23 | 苏州赛谱仪器有限公司 | Continuous injection chromatography device |
| CN106769362A (en) * | 2017-02-08 | 2017-05-31 | 中国环境科学研究院 | A kind of full-automatic adsorption experimental apparatus |
| CN109320603A (en) * | 2018-12-17 | 2019-02-12 | 杭州奕安济世生物药业有限公司 | A kind of system and method for serialization purification antibody |
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| WO2007109157A2 (en) * | 2006-03-17 | 2007-09-27 | Waters Investments Limited | Solvent delivery system for liquid chromatography that maintains fluid integrity and pre-forms gradients |
| CN201133904Y (en) * | 2008-01-01 | 2008-10-15 | 朱海波 | Negative-pressure liquid phase chromatograph |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102008837A (en) * | 2010-09-30 | 2011-04-13 | 中国科学院昆明植物研究所 | Separating preparative chromatography apparatus comprising serial preparative chromatography column |
| CN102441294A (en) * | 2010-09-30 | 2012-05-09 | 中国科学院昆明植物研究所 | Separation and preparation chromatograph containing series static axial compression (SAC) preparative chromatographic columns |
| CN102441295A (en) * | 2010-09-30 | 2012-05-09 | 中国科学院昆明植物研究所 | Separation and preparation chromatograph containing series dynamic axial compression (DAC) preparative chromatographic columns |
| CN102008837B (en) * | 2010-09-30 | 2012-09-05 | 中国科学院昆明植物研究所 | Separating preparative chromatography apparatus comprising serial preparative chromatography column |
| CN102172516A (en) * | 2011-03-28 | 2011-09-07 | 天津博纳艾杰尔科技有限公司 | Novel resin-coated silica-gel adsorption material |
| CN102172516B (en) * | 2011-03-28 | 2013-05-15 | 天津博纳艾杰尔科技有限公司 | Novel resin-coated silica-gel adsorption material |
| CN104407083A (en) * | 2014-11-28 | 2015-03-11 | 天津博纳艾杰尔科技有限公司 | Full-automatic separation system and application of full-automatic separation system in separation of polar components in edible oil |
| CN104407083B (en) * | 2014-11-28 | 2016-08-31 | 天津博纳艾杰尔科技有限公司 | Full-automatic piece-rate system and the application in edible oil polar compound separates thereof |
| WO2017028357A1 (en) * | 2015-08-18 | 2017-02-23 | 苏州赛谱仪器有限公司 | Continuous injection chromatography device |
| CN106769362A (en) * | 2017-02-08 | 2017-05-31 | 中国环境科学研究院 | A kind of full-automatic adsorption experimental apparatus |
| CN106769362B (en) * | 2017-02-08 | 2018-06-29 | 中国环境科学研究院 | A kind of full-automatic adsorption experimental apparatus |
| CN109320603A (en) * | 2018-12-17 | 2019-02-12 | 杭州奕安济世生物药业有限公司 | A kind of system and method for serialization purification antibody |
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Addressee: Xu Ping Document name: Notification to Make Rectification |
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| C06 | Publication | ||
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