CN100390536C - High flux protein multidemension array chromatogram separating system - Google Patents
High flux protein multidemension array chromatogram separating system Download PDFInfo
- Publication number
- CN100390536C CN100390536C CNB2004100170371A CN200410017037A CN100390536C CN 100390536 C CN100390536 C CN 100390536C CN B2004100170371 A CNB2004100170371 A CN B2004100170371A CN 200410017037 A CN200410017037 A CN 200410017037A CN 100390536 C CN100390536 C CN 100390536C
- Authority
- CN
- China
- Prior art keywords
- array
- chromatogram
- column
- chromatographic column
- dimension
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
The present invention relates to a high flux protein multidimensional chromatogram separating system. In the prior art, the methods of the chromatographic analysis of proteome are only limited to single channel one-dimensional or two-dimensional chromatographic methods and have the defects of long analysis cycle, complicated operation, etc. The present invention uses a multichannel chromatogram array for the second dimension chromatographic separation methods on the basis of first dimension chromatographic separation, thereby causing analysis efficiency to be greatly enhanced; the multichannel chromatogram array can extend 2 to 50 chromatogram passages and can be connected with a first dimension chromatogram through multichannel interfaces; the multichannel chromatogram array can complete the separation process of enrichment column by column and gradient elution and provide an effective method for proteome sample analysis. The members used in the method of the present invention are integrated into a whole, are used with substrate auxiliary laser ionization mass spectrums together and can realize an automatic proteome color-mass instrument.
Description
Technical field
The present invention is a high throughput protein multi-dimension array chromatographic fractionation system, and the separation method of this system of utilization.
Background technology
Proteomics is one of present scientific research hottest point field.Its research essence is on cellular level protein to be separated on a large scale and analyze.Different with the genome of static state, protein has complicated posttranslational modification, is various, dynamic in intracellular expression.Because the high complexity of protein group sample, the rise of proteomics and development have proposed new demand and challenge to existing analytical technology.Development high flux, high score become the bottleneck of current proteomics research from the technology platform of ability.
The core technology of current proteome research in the world is two dimensional gel electrophore-sis-mass-spectrometric technique.Separate the most frequently used instrument although two dimensional gel electrophore-sis is a protein group, also there are many limitations this technology in this on analysis throughput, sensitivity, resolution and automaticity, is difficult to satisfy the needs that carry out proteomics research on a large scale.Studying new technology alternative or additional two dimensional gel electrophore-sis becomes the main target of development proteome research technology.Carried out many exploration work both at home and abroad in this respect, multiple separation method is used widely and is developed, as: high performance liquid chromatography, Capillary Electrophoresis and multidimensional liquid chromatography etc.Generally believed it is the proteome research technology of tool potentiality because the high complexity of protein group sample, two dimension and even multidimensional are separated combining of coupling technique and mass spectrum authenticate technology, paid close attention to widely in the proteome research field and pay attention to.
Both at home and abroad in the research that is only limited to single channel one peacekeeping two dimension chromatographic system aspect the chromatogram high efficiency separation of protein, hyperchannel multi-dimensional chromatograph piece-rate system (more than 4 passages) does not still have report at present.Although two-dimentional chromatographic system is in existing commercial instrument release last year, but also there is restriction greatly in it on analysis throughput, a sample needs repeatedly the wash-out circulation of (polyhalinicum gradient and reverse phase gradient), prolonged analysis time greatly, and the pre-classification of warp of general analysis is handled the protein group sample of simplifying and is needed 2-3 days.
Summary of the invention
The objective of the invention is to obtain a kind of high throughput protein multi-dimension array chromatographic fractionation system.
The objective of the invention is to obtain to carry out the method for high throughput protein multi-dimension array chromatographic resolution with said system.
Analytic system of the present invention realizes with following mode:
System is made of parallel array chromatographic column (1), single chromatographic column (4) and hyperchannel spot sample device, it is characterized in that array chromatographic column (1) is connected with single chromatographic column (4) by hyperchannel chromatogram connector (5), array chromatographic column tail all has post switch (6), chromatogram pump I (3), single chromatographic column (4), hyperchannel chromatogram connector (5), chromatogram pump II (8) connection of contacting successively, the multi-dimensional chromatograph array separating part of above construction system; Miniature three-way mixing chamber (10), syringe pump (9), point sample kapillary (2), point template (11) and the three-dimensional hyperchannel spot sample device of mobile platform (12) construction system automatically, on the miniature three-way mixing chamber (15), (17) two-port is connected with point sample kapillary (2) with array chromatographic column column outlet respectively down, side ports (16) is connected with the syringe pump (9) of carrying matrix solution, miniature three-way mixing chamber (15) is corresponding one by one with the array post, under the point sample kapillary (2) point template (11), point template places on the three-dimensional mobile platform (12) automatically, and above-mentioned each parts are integrated in one.
Testing sample is at first gone up wash-out in single chromatographic column (4), the wash-out effluent is pursued on each column cap (7) that post is enriched to the array chromatographic column successively by hyperchannel chromatogram connector (5) utilization array post switch (6), the array chromatographic column is carried out chromatography eluant by chromatogram pump II (8), meanwhile by the hyperchannel spot sample device, the wash-out effluent of each passage of chromatogram array passes through syringe pump (9) on-line mixing in corresponding miniature three-way mixing chamber (10) with matrix solution respectively, eluate and substrate mixture are dispersed the place on point template (11) by setting spacing, and sample is for substance assistant laser desorpted ionized Mass Spectrometer Method.
Therefore analytic system of the present invention is made of the bidimensional chromatogram, and the first dimension chromatographic resolution is utilized a root chromatogram column, can adopt ion-exchange (or volume exclusion) chromatographic column; The second dimension chromatographic resolution is utilized many root chromatogram columns (50 of as many as), can adopt reverse-phase chromatographic column.Two piece-rate systems connect by multi-channel parallel formula multi-dimensional chromatograph interface.
The first dimension separation realizes (each component only needs a few minutes) fast by ion-exchange (or volume exclusion) chromatogram, elution fraction is switched on the column cap or enriching column that is enriched in the second dimension chromatogram array reversed-phase column in turn respectively through hyperchannel chromatogram interface or multi-position valve, and these a plurality of elution fractions are finished by wash-out of mode of parallel chromatogram in the separation of second dimension.The analysis throughput of this two-dimensional array chromatographic system can be expressed as: analysis throughput (2D-LC array)=N
1DfractionXT
1D+ 1xT
2D, and the analysis throughput of common single channel bidimensional chromatographic system is: analysis throughput (2D-LC)=N
1DfractionXT
1D+
N 1Dfraction XT
2D, (N is that sample is through the first dimension group number that wash-out divided, the N=2-50 usually; T
1DBe the elution time of first dimension, be generally a few minutes; T
2DBe the elution time of second dimension, be generally 1-3 hour), this shows that utilization array chromatogram can greatly improve the sample analysis flux of system as second dimension of system.The outflow component of the second dimension multichannel array chromatography eluant is collected in by miniature point sample system carries out TOF/TOF or the Q/TOF mass spectrum is identified on the MALDI plate.
With the corresponding miniature three-way mixing chamber of array chromatographic column array, point sample capillary array, kapillary geometrical clamp, carry the miniature point sample part of matrix capillary array construction system.Miniature three-way mixing chamber array is by constituting with the long-pending mixing chamber of the corresponding microbody of array chromatographic column, and the long-pending mixing chamber of each microbody has three ports: upper port, lower port and side ports.Each upper port is connected with the second dimension chromatographic column respectively, and lower port is connected with the point sample kapillary respectively, and side ports is connected with the matrix discharge pump.Be carved with equally spaced conduit in the kapillary geometrical clamp, be used for fixing the point sample capillary array.
Among the present invention, array chromatographic column column cap is connected with enriching column (14), and post switch (6) is connected the enriching column tail.
Among the present invention, the parallel array chromatographic column is the 2-50 root chromatogram column.
Among the present invention, array chromatographic column (1) is connected with single chromatographic column (4) by multi-position valve (13).
Among the present invention, the parallel array chromatographic column is to carry out chromatography eluant simultaneously after the enrichment of single chromatographic column eluent.
Among the present invention, the wash-out effluent that switches to the array chromatographic column is enriched in (14) on each enriching column.
Among the present invention, system and substance assistant laser desorpted ionized mass spectrometry.
The present invention has obtained following good result:
A plurality of passages carry out the second dimension chromatographic resolution simultaneously, realized the superelevation analysis throughput, utilization multi-channel parallel formula multi-dimensional chromatograph interface, make system obtain the multidimensional handoff technique, and a plurality of chromatographic columns can be separated simultaneously, multidimensional separated flow part is transferred to by a model machine carries out TOF/TOF or Q/TOF mass spectrum on the MALDI plate and identify, can realize the protein high flux, fast, efficient analysis.
The present invention is main separation platform with chromatogram one chromatogram, give full play to that chromatogram separating capacity is strong, clastotype is many, be easy to identify the mode coupling, be easy to advantage such as robotization with mass spectrum, structure is greatly improved the protein technique platform based on the array protein group multidimensional isolation technics platform that the multidimensional high efficiency chromatography separates on separation efficiency and analysis throughput.Analyze a sample with 2D-gel at present and need several days time at least, single channel 2D-LC needs tens hours.Because the resolution of two-dimentional LC separation platform and analysis throughput mainly by the decision that separates of second dimension, are analyzed simultaneously at a plurality of passages of the second dimension employing, can greatly improve the analysis throughput of total system.In addition with the two row chromatographic columns wash-out that hockets, having the more sufficient time carries out the second dimension gradient separations, thereby further improves the resolution of 2D-LC.Array multidimensional platform with a plurality of passages structures, its analysis throughput is with firm commercial single channel 2D-LC ratio, the raising of tens times and even tens times will be had, the analysis that breaks through proteins/peptides is in resolution, sensitivity, the limitation of aspect such as analysis throughput particularly for the research of protein group provides effective technical means more, thereby further promotes the flow of research in this field.
Description of drawings
Fig. 1 is an analytic system structural representation of the present invention.
Fig. 2 is the system architecture synoptic diagram of system employs multi-position valve 13 of the present invention, enriching column 14.
Fig. 3 is hyperchannel chromatogram array interface figure of the present invention.
Fig. 4 is a miniature three-way mixing chamber structural drawing of the present invention.
1 is the array chromatographic column among the above-mentioned figure, the 2nd, and point sample kapillary, the 3rd, chromatogram pump I, the 4th, single chromatographic column, the 5th, hyperchannel interface, the 6th, post switch, the 7th, the column cap of array chromatographic column, the 8th, chromatogram pump II, the 9th, syringe pump, the 10th, miniature three-way mixing chamber, the 11st, point template, the 12nd, three-dimensional mobile platform automatically, the 13rd, multi-position valve, the 14th, enriching column, 15,17 is respectively miniature three-way mixing chamber upper/lower terminal mouth, the 16th, horizontal port.
Embodiment
One, the foundation of high throughput protein multi-dimension array chromatographic fractionation system
1. the making of hyperchannel chromatogram interface:
Polyetheretherketone (PEEK) kapillary that to get a long 5cm internal diameter be 500 μ m gets out 30 of the apertures that diameter is 380 μ m in a PEEK side capillaceous with the drill bit of 380 μ m, and the spacing of Kong Yukong is 1mm, and the chip in the flush away PEEK pipe also dries up it with nitrogen.Insert the quartz capillary that external diameter is 480 μ m along the PEEK kapillary, with 30 long 5cm external diameters quartz capillary (75 μ m i.d. that are 380 μ m; 380 μ m o.d.) insert respectively in the aperture and offset, extract the built-in kapillary of PEEK central passage, subsequently the fixing with glue all around of quartz capillary and PEEK kapillary intersection, as Fig. 3 with built-in kapillary.
2. the making of hyperchannel spot sample device:
The hyperchannel spot sample device amasss the mixing chamber array by the threeway microbody, the point sample capillary array, and the kapillary geometrical clamp carries the matrix capillary array to constitute.The long-pending mixing chamber array of microbody is made of the long-pending mixing chamber of 30 microbodys, and the long-pending mixing chamber structure of microbody as shown in Figure 4.
3. the foundation of array bidimensional chromatographic fractionation system:
First Wesy, one radical ion exchange chromatography post (3cm * 320 μ m) links to each other with the end that the PEEK pipe of injector and hyperchannel chromatogram interface exports respectively, as shown in Figure 2.The other end of the PEEK of hyperchannel chromatogram interface pipe outlet links to each other with gradient pump, on the interface outlet of quartz capillary link to each other with reverse-phase chromatographic column respectively (250 μ m i.d * 25cm, 5 μ m, C8); The outlet of chromatogram array links to each other with the PEEK switch respectively, links to each other with the upper inlet of the long-pending mixing chamber array of microbody with the following outlet of a quartz capillary with switch.10 horizontal channels link to each other with 10 syringe pumps respectively (or with a syringe pump be connected by part flow arrangement).Hyperchannel point sample system is placed the top 10cm of Linomat (or automatic three-dimensional mobile platform), and the MALDI sample panel is placed on the three-dimensional mobile platform.
Two. carry out the protein group sample analysis with building high throughput protein multi-dimension array chromatographic fractionation system
The holoprotein extract of cell or tissue is in 100 ℃ of water-bath heat denatured.Reduce to room temperature, with trypsin digestion.
After institute built bidimensional chromatogram array piece-rate system balance, (switch was outwarded winding after being designated as post post a), and other is closed with the root chromatogram column in the chromatogram array.The proteolysis peptide section potpourri of hydrolysis all is written in the bidimensional chromatogram array piece-rate system (SCX on the first dimension ion-exchange chromatography through injector, 320 μ m i.d * 3cm), with chromatogram pump I is moving phase with 5%ACN/0.1%TFA, the flow wash-out of 3 μ l/min, the peptide section component under the wash-out is transferred to post a and is enriched in column cap.Close switch behind the post a post, open chromatographic column (the being designated as post b) switch behind the post a, advance the 10mM ammonium acetate solution with injector, the peptide section component that this salt solusion elutes is enriched in post b column cap through the bidimensional interface.As above same operation is respectively with 50,100,150,200,250,300,500 and the peptide composition of 1000mM ammonium acetate wash-out be enriched in the column cap of 8 reversed-phase columns of back.Close chromatogram pump I subsequently, with switch opens behind all posts, trigger chromatogram pump II, with moving phase (mobile phase A: 5%ACN/0.1%TFA and Mobile phase B: 80%ACN/0.1%TFA) elution chromatography array.
Syringe pump (LC Packing, Amsterdam, The Netherlands) is connected with the shunt of the long-pending mixing chamber array of microbody by 10 paths.In chromatogram array wash-out, matrix solution is imported respectively in each mixing chamber by syringe pump.The effluent of chromatogram array wash-out mixes in mixing chamber with matrix solution, and potpourri was put on the MALDI sample panel automatically with the time interval that is provided with.
With AB 4700 Proteomics Analyzer each sample spot is carried out the mensuration of MALDI/MS and MALDI/MS/MS at last.With GPS software the whole MS/MS data of gained are handled, and in Protein Data Bank, searched for, thereby obtain the qualification result of protein.
Claims (8)
1. high throughput protein multi-dimension array chromatographic fractionation system, system is made of parallel array chromatographic column (1), single chromatographic column (4) and hyperchannel spot sample device, it is characterized in that array chromatographic column (1) is connected with single chromatographic column (4) by hyperchannel chromatogram connector (5), array chromatographic column post tail all has post switch (6), chromatogram pump I (3), single chromatographic column (4), hyperchannel chromatogram connector (5), chromatogram pump II (8) connection of contacting successively, the multi-dimensional chromatograph array separating part of above construction system; Miniature three-way mixing chamber (10), syringe pump (9), point sample kapillary (2), point template (11) and the three-dimensional hyperchannel spot sample device of mobile platform (12) construction system automatically, miniature three-way mixing chamber upper port (15), lower port (17) two-port is connected with point sample kapillary (2) with array chromatographic column column outlet respectively, side ports (16) is connected with the syringe pump (9) of carrying matrix solution, miniature three-way mixing chamber (10) is corresponding one by one with the array chromatographic column, under the point sample kapillary (2) point template (11), point template places on the three-dimensional mobile platform (12) automatically, and above-mentioned each parts are integrated in one.
2. high flux multi-dimension array chromatographic fractionation system according to claim 1 is characterized in that array chromatographic column column cap is connected with enriching column (14), and post switch (6) is connected the enriching column tail.
3. high throughput protein multi-dimension array chromatographic fractionation system according to claim 1 is characterized in that the parallel array chromatographic column is the 2-50 root chromatogram column.
4. high throughput protein multi-dimension array chromatographic fractionation system according to claim 1 is characterized in that array chromatographic column (1) is connected with single chromatographic column (4) by multi-position valve (13).
5. carry out the method for protein multi-dimension array chromatographic resolution with the described system of claim 1, it is characterized in that testing sample is at first at the last wash-out of single chromatographic column (4), the wash-out effluent is pursued on each column cap (7) that post is enriched to the array chromatographic column successively by hyperchannel chromatogram connector (5) utilization array post switch (6), the array chromatographic column is carried out chromatography eluant by chromatogram pump II (8), meanwhile by the hyperchannel spot sample device, the wash-out effluent of each passage of chromatogram array passes through syringe pump (9) on-line mixing in corresponding miniature three-way mixing chamber (10) with matrix solution respectively, eluate and substrate mixture are dispersed the place on point template (11) by setting spacing, and sample is for substance assistant laser desorpted ionized Mass Spectrometer Method.
6. the separation method of high throughput protein multi-dimension array chromatographic fractionation system according to claim 5 is characterized in that the parallel array chromatographic column is to carry out chromatography eluant simultaneously.
7. the separation method of high throughput protein multi-dimension array chromatographic fractionation system according to claim 5 is characterized in that the wash-out effluent that switches to the array chromatographic column is enriched on each enriching column (14).
8. high throughput protein multi-dimension array chromatographic fractionation system according to claim 1 and 2 is characterized in that this system and substance assistant laser desorpted ionized mass spectrometry.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100170371A CN100390536C (en) | 2004-03-18 | 2004-03-18 | High flux protein multidemension array chromatogram separating system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100170371A CN100390536C (en) | 2004-03-18 | 2004-03-18 | High flux protein multidemension array chromatogram separating system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1563977A CN1563977A (en) | 2005-01-12 |
| CN100390536C true CN100390536C (en) | 2008-05-28 |
Family
ID=34478739
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100170371A Expired - Fee Related CN100390536C (en) | 2004-03-18 | 2004-03-18 | High flux protein multidemension array chromatogram separating system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100390536C (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20070017808A1 (en) * | 2005-05-27 | 2007-01-25 | Intel Corporation | Linear valve-coupled two-dimensional separation device and separation matrix and method |
| CN103063753B (en) * | 2012-08-31 | 2015-07-29 | 西安奥岚科技开发有限责任公司 | For multidimensional liquid chromatographic separation system and the separation method of Protein Separation |
| CN103212217B (en) * | 2013-04-20 | 2015-05-13 | 复旦大学 | Two-dimensional conventional column array type chromatographic separation system and method for removing high-abundance proteins |
| CN103323544B (en) * | 2013-05-28 | 2014-10-22 | 同济大学 | On-line pre-separating and separating analysis method for whole protein and liquid chromatogram system thereof |
| CN105891363A (en) * | 2016-04-08 | 2016-08-24 | 厦门大学 | Multi-probe liquid chromatography tandem mass spectrometry point plate device |
| CN113533558A (en) * | 2021-06-28 | 2021-10-22 | 复旦大学 | Array type two-dimensional liquid chromatography separation system and separation method for separating protein compound |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5240577A (en) * | 1990-11-13 | 1993-08-31 | University Of North Carolina At Chapel Hill | Two-dimensional high-performance liquid chromatography/capillary electrophoresis |
| EP1235634A1 (en) * | 1999-11-16 | 2002-09-04 | James T. Champagne | Solution based two-dimensional separation and detection of amphoteric substances |
| US20030089663A1 (en) * | 1999-04-02 | 2003-05-15 | Miroslav Petro | Methods and apparatus for characterization of polymers using multi-dimensional liquid chromatography with parallel second-dimension sampling |
| US6581441B1 (en) * | 2002-02-01 | 2003-06-24 | Perseptive Biosystems, Inc. | Capillary column chromatography process and system |
-
2004
- 2004-03-18 CN CNB2004100170371A patent/CN100390536C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5240577A (en) * | 1990-11-13 | 1993-08-31 | University Of North Carolina At Chapel Hill | Two-dimensional high-performance liquid chromatography/capillary electrophoresis |
| US20030089663A1 (en) * | 1999-04-02 | 2003-05-15 | Miroslav Petro | Methods and apparatus for characterization of polymers using multi-dimensional liquid chromatography with parallel second-dimension sampling |
| EP1235634A1 (en) * | 1999-11-16 | 2002-09-04 | James T. Champagne | Solution based two-dimensional separation and detection of amphoteric substances |
| US6581441B1 (en) * | 2002-02-01 | 2003-06-24 | Perseptive Biosystems, Inc. | Capillary column chromatography process and system |
Non-Patent Citations (2)
| Title |
|---|
| 多维立体分离分析技术面临的挑战性问题及对应策略. 张玉奎，张维冰，张丽华.色谱，第21卷第4期. 2003 |
| 多维立体分离分析技术面临的挑战性问题及对应策略. 张玉奎，张维冰，张丽华.色谱，第21卷第4期. 2003 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1563977A (en) | 2005-01-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8074494B2 (en) | Plugged flow 2D chromatography | |
| Simpson et al. | Combining capillary electrophoresis with mass spectrometry for applications in proteomics | |
| US20080156080A1 (en) | Methods and systems for multidimensional concentration and separation of biomolecules using capillary isotachophoresis | |
| CN108562678B (en) | Three-dimensional liquid chromatographic separation system based on full online detection of same detector | |
| Hata et al. | Two-dimensional HPLC on-line analysis of phosphopeptides using titania and monolithic columns | |
| CN202421144U (en) | Two-dimensional gas-phase chromatograph-double-mass spectrum central cutting device | |
| CN100390536C (en) | High flux protein multidemension array chromatogram separating system | |
| CN109557219A (en) | A kind of multidimensional liquid chromatographic separation system | |
| KR101617615B1 (en) | Dual online liquid chromatography apparatus and control method of the same | |
| CN105241984B (en) | Array multidimensional liquid chromatogram column system | |
| CN207586196U (en) | The multidimensional liquid chromatographic separation system of full on-line checking based on same detector | |
| CN103212217B (en) | Two-dimensional conventional column array type chromatographic separation system and method for removing high-abundance proteins | |
| Gao et al. | Development of multidimensional liquid chromatography and application in proteomic analysis | |
| Capriotti et al. | A multidimensional liquid chromatography–tandem mass spectrometry platform to improve protein identification in high-throughput shotgun proteomics | |
| JP2004101477A (en) | Two-dimensional high performance liquid chromatographic device and protein analyzer using the same | |
| CA2975027A1 (en) | Means and methods for minimizing swept and dead volumes in chromatographic applications | |
| WO2006062471B1 (en) | A methods and interfaces for single and multidimentional separations for characterization and/or identification of molecules by mass spectrometry | |
| US8518346B1 (en) | Multidimensional bioseparation with modular microfluidics | |
| CN1785476A (en) | Full bidimensional capillary liquid chromatography separation system | |
| CN207198107U (en) | A kind of cascade Coupled columns mass spectrometer system based on valve | |
| Liu et al. | Multidimensional capillary array liquid chromatography and matrix-assisted laser desorption/ionization tandem mass spectrometry for high-throughput proteomic analysis | |
| CN206132715U (en) | Development of collection separation method , online separation enrichment is in crude drug of an organic whole two dimension preparative chromatograph | |
| CN114235994A (en) | Non-continuous sample grading and cascading device and double-online multi-dimensional liquid chromatography system with same | |
| CN214310345U (en) | Separation device and analysis system for separating high-abundance and low-abundance proteins of tobacco through liquid chromatography | |
| CN205091306U (en) | Array multidimension liquid chromatogram column system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20080528 Termination date: 20110318 |