CN103412029A - Flat plate electrochromatography amino acid separating device based on chip level and use method thereof - Google Patents
Flat plate electrochromatography amino acid separating device based on chip level and use method thereof Download PDFInfo
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- CN103412029A CN103412029A CN2013102614681A CN201310261468A CN103412029A CN 103412029 A CN103412029 A CN 103412029A CN 2013102614681 A CN2013102614681 A CN 2013102614681A CN 201310261468 A CN201310261468 A CN 201310261468A CN 103412029 A CN103412029 A CN 103412029A
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- amino acid
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- silica gel
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Abstract
The invention relates to a flat plate electrochromatography amino acid separating device based on a chip level and a use method thereof. The flat plate electrochromatography amino acid separating device comprise a CCD (Charge Coupled Device) camera, a fluorescence microscope imaging system, electrode modules, a silica gel chromatography plate, a bottom supporting component and a power supply, wherein the electrode modules are horizontally arranged at two ends of the silica gel chromatography plate; the bottom supporting component is used for fixing the chromatography plate and the electrode modules to form a carrier for separation analysis; the CCD camera and the fluorescence microscope imaging system are arranged right above the integral device; the power supply is connected into the electrode modules through power supply wires. The device has the beneficial effects of being simple and low in cost and portable in size; experiments prove that the method is stable, reliable and effective, and extremely high in compatibility; the whole process becomes the two-dimensional separation as electric field action is introduced in the originally one-dimensional chromatography process, and the chromatographic separation principle and electrophoretic separation act in separation and analysis of amino acids at the same time.
Description
[technical field]
The present invention relates to amino acid tripping device technical field, specifically, is a kind of dull and stereotyped electrochromatography amino acid tripping device and using method thereof based on chip level.
[background technology]
Amino acid is to form bioprotein and, with the relevant base substance of vital movement, with biological vital movement, close relationship is arranged.As the amino acid of the base unit that forms protein molecule, the detection to protein sequence in genetic engineering has extremely important research status.(Henikoff，S.；Henikoff，J.；1992，Proc.Natal.Acad.Sci.USA，89，10915-10919)。Since amino acid whose importance was by extensive concern, scholar both domestic and external dropped into a large amount of scientific research and concern for amino acid whose compartment analysis, and has obtained numerous progress.Such as near-infrared method is measured amino acid (Zhao Chen; Qu Haibin; 2004, spectroscopy and spectral analysis, 24), high efficiency liquid phase method (Shi Chunzhen; Zhang Hongman; 2012, analytical chemistry, 4), (Jiang Tao such as automatic amino acid analyzer method for quick; Feng Yongjian; 2012, chemistry application and research, 7) etc.Wherein most methods and instrument are all to rely on business-like large-scale plant to analyze and detect, for the instrument requirement, and testing environment, many conditions such as sample purity have comparatively harsh requirement.The method complex operation even had, testing cost is higher.Therefore, be necessary to propose a kind of based on chip technology with low cost, the separation of Simple fast and the reliable method of analysis of amino acid.
Some analytical approachs certain specific amino acid in can the mensuration complex sample of Simple fast has been arranged at present, as (the Schulta C.L. of the coupling technique by Capillary Electrophoresis, Moini M.2003, Anal.Chem., 75,1508-1513) and mass-spectrometric technique (Soga T., Heiger D.N., 2000, Anal.Chem., 75,1508-1513).Although these methods can Accurate Determining amino acid, their selectivity and practicality all can be subject to the interference of one or more external factors, to amino acid whose detection flux, are not therefore very high.In addition, these methods often need expensive instrument and equipment, complicated operation, and minute is longer.
Chip technology is through fast development, widespread use and the numerous analytical chemistry field of nearly more than ten years.Such as isoelectric focusing IEF chip (Cui H.C., Keisuke H.2005, Anal.Chem., 77,1303-1309) and Capillary Electrophoresis CE chip (Carlo S.E., Gerard J.M.B., 1997, Anal.Chem., 69,3451-3457).Above-mentioned chip technology is for analysis capillaceous and detection, harsher and single-minded for the requirement of detecting device, such as fluoroscopic examination or ultraviolet detection.All need to carry out the collection of signal and the conversion of data message by photomultiplier or special CCD.And also there is larger blank in chip field for the research of the analysis chip in thin layer chromatography field.
[summary of the invention]
The object of the invention is to overcome the deficiencies in the prior art, a kind of dull and stereotyped electrochromatography amino acid tripping device and using method thereof based on chip level is provided.
The objective of the invention is to be achieved through the following technical solutions:
A kind of dull and stereotyped electrochromatography amino acid tripping device based on chip level, it comprises CCD camera and fluorescent microscope imaging system, electrode module, silica gel column chromatography plate, bottom support parts and power supply; Electrode module is placed horizontally at the two ends of silica gel column chromatography plate, and the bottom support parts are used for fixedly chromatoplate and electrode module, forms the carrier of compartment analysis; CCD camera and fluorescent microscope imaging system are positioned over directly over single unit system, and power supply is connected in electrode module by power circuit.
Described CCD camera and fluorescent microscope imaging system, pixel are 300～6,000,000, and the fluorogram chip resolution is 300～1000DPI, and the CCD imaging region is 1～4 square centimeter, and the enlargement factor of microscope stage is 50～500 times.
The specification that described electrode module comprises the electrode holder of PMMA polymethylmethacrylate material making is 2～3 square centimeters, and the specification of the electrode ports that scribe at two ends is 1～2 square millimeter, and the specification of metal platinum wire electrode is 30～60 millimeters.
The specification that described silica gel column chromatography plate comprises glass plate is long 30～40 square centimeters, and the silica-gel coating thickness specification of solid chromatographic silica gel is 0.1～0.2 millimeters thick, and the silica gel particle degree is 200～300 orders.
Described bottom support parts are the support baseboard that PMMA polymethylmethacrylate material is made, and for fixed electorde module and chromatoplate, specification is 30～40 square centimeters.
Described power supply is the DC power supply apparatus of 0～600V.
A kind of using method of the dull and stereotyped electrochromatography amino acid tripping device based on chip level, its concrete steps are, the electrode module of described built-in platinum wire electrode is placed horizontally at the two ends of glass silica gel chromatoplate, the bottom support parts are used for fixedly chromatoplate and electrode module, form the carrier of compartment analysis; Sample is entered to the loading zone of silica gel plate by a loading; CCD camera and fluorescent microscope imaging system are positioned over directly over single unit system, and power supply is connected in electrode module by power circuit; In use procedure, the lower end of silica gel column chromatography plate is immersed in the buffer solution that 5～10mm is dark, in the chromatography effect, introduced the purpose that electric field action reaches two dimensional separation.
The device of this chip level can for two dimensional separation is multiple can fluorescently-labeled amino acid.
Compared with prior art, good effect of the present invention is:
The first, simple and cheap installation cost, plant bulk that can be portable.
Second and prove that by experiment the method is stable, reliable, effectively, and compatible high.
Three, by the chromatography process at the script one dimension, introducing electric field action, make whole process become two-dimentional separation.Chromatographic resolution principle and electrophoretic separation jointly and are simultaneously acted on and amino acid whosely separate and analyze.
[accompanying drawing explanation]
Fig. 1 is principle schematic of the present invention;
Fig. 2 is structure drawing of device of the present invention;
Fig. 3 is separating resulting figure of the present invention, No. 1 caption be the mobile band of the sample under added electric field not; No. 2 captions be advanced row buffering liquid syphonic effect, add subsequently electric field, sample is in the electrophoresis mobile status of silica gel plate; No. 3 captions be under the state that acts on simultaneously of siphon and electric field, the electrophoretic band state of component to be separated; No. 4 picture is repeated experiment, and condition is identical with No. 3 pictures.
Being labeled as in accompanying drawing: 1CCD camera and fluorescent microscope imaging system, 2 kapillary loading parts, 3 electrode modules, 4 silica gel column chromatography plates, 5 sample loading ports, 6 bottom support parts.
[embodiment]
A kind of dull and stereotyped electrochromatography amino acid tripping device based on chip level of the present invention and the embodiment of using method thereof below are provided.
A kind of dull and stereotyped electrochromatography amino acid tripping device based on chip level, it comprises CCD camera and fluorescent microscope imaging system, electrode module, silica gel column chromatography plate, bottom support parts and power supply; Electrode module is placed horizontally at the two ends of silica gel column chromatography plate, and the bottom support parts are used for fixedly chromatoplate and electrode module, forms the carrier of compartment analysis; CCD camera and fluorescent microscope imaging system are positioned over directly over single unit system, and power supply is connected in electrode module by power circuit.
Described CCD camera and fluorescent microscope imaging system, pixel are 300～6,000,000, and the fluorogram chip resolution is 300～1000DPI, and the CCD imaging region is 1～4 square centimeter, and the enlargement factor of microscope stage is 50～500 times.
The specification that described electrode module comprises the electrode holder of PMMA polymethylmethacrylate material making is 2～3 square centimeters, and the specification of the electrode ports that scribe at two ends is 1～2 square millimeter, and the specification of metal platinum wire electrode is 30～60 millimeters.
The specification that described silica gel column chromatography plate comprises glass plate is long 30～40 square centimeters, and the silica-gel coating thickness specification of solid chromatographic silica gel is 0.1～0.2 millimeters thick, and the silica gel particle degree is 200～300 orders.
Described bottom support parts are the support baseboard that PMMA polymethylmethacrylate material is made, and for fixed electorde module and chromatoplate, specification is 30～40 square centimeters.
Described power supply is the DC power supply apparatus of 0～600V.
A kind of using method of the dull and stereotyped electrochromatography amino acid tripping device based on chip level, its concrete steps are, the electrode module of described built-in platinum wire electrode is placed horizontally at the two ends of glass silica gel chromatoplate, the bottom support parts are used for fixedly chromatoplate and electrode module, form the carrier of compartment analysis; Sample is entered to the loading zone of silica gel plate by a loading; CCD camera and fluorescent microscope imaging system are positioned over directly over single unit system, and power supply is connected in electrode module by power circuit; In use procedure, the lower end of silica gel column chromatography plate is immersed in the buffer solution that 5～10mm is dark, in the chromatography effect, introduced the purpose that electric field action reaches two dimensional separation.
As shown in Figure 1, the present embodiment comprises: CCD camera 1, kapillary loading parts 2, electrode module 3, silica gel column chromatography plate 4, sample loading port 5, bottom support parts 6.
As shown in Figure 2, in separation principle figure, show the length of electrode module; The direction of damping fluid syphonic effect has been described in the black dotted lines frame; The difference of the electrophoresis direction of the colored arrow various charge specieses of simulation under electric field action; The top mark attribute of extra electric field.
As shown in Figure 3, No. 1 caption is the mobile band of the sample under added electric field not; No. 2 captions be advanced row buffering liquid syphonic effect, add subsequently electric field, sample is in the electrophoresis mobile status of silica gel plate; No. 3 captions be under the state that acts on simultaneously of siphon and electric field, the electrophoretic band state of component to be separated; No. 4 picture is repeated experiment, and condition is identical with No. 3 pictures.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, without departing from the inventive concept of the premise; can also make some improvements and modifications, these improvements and modifications also should be considered within the scope of protection of the present invention.
Claims (8)
1. dull and stereotyped electrochromatography amino acid tripping device based on chip level, it comprises CCD camera and fluorescent microscope imaging system, electrode module, silica gel column chromatography plate, bottom support parts and power supply; It is characterized in that, electrode module is placed horizontally at the two ends of silica gel column chromatography plate, and the bottom support parts are used for fixedly chromatoplate and electrode module, forms the carrier of compartment analysis; CCD camera and fluorescent microscope imaging system are positioned over directly over single unit system, and power supply is connected in electrode module by power circuit.
2. a kind of dull and stereotyped electrochromatography amino acid tripping device based on chip level as claimed in claim 1, it is characterized in that, described CCD camera and fluorescent microscope imaging system, pixel is 300～6,000,000, the fluorogram chip resolution is 300～1000DPI, the CCD imaging region is 1～4 square centimeter, and the enlargement factor of microscope stage is 50～500 times.
3. a kind of dull and stereotyped electrochromatography amino acid tripping device based on chip level as claimed in claim 1, it is characterized in that, the specification that described electrode module comprises the electrode holder of PMMA polymethylmethacrylate material making is 2～3 square centimeters, the specification of the electrode ports that scribe at two ends is 1～2 square millimeter, and the specification of metal platinum wire electrode is 30～60 millimeters.
4. a kind of dull and stereotyped electrochromatography amino acid tripping device based on chip level as claimed in claim 1, it is characterized in that, the specification that described silica gel column chromatography plate comprises glass plate is long 30～40 square centimeters, the silica-gel coating thickness specification of solid chromatographic silica gel is 0.1～0.2 millimeters thick, and the silica gel particle degree is 200～300 orders.
5. a kind of dull and stereotyped electrochromatography amino acid tripping device based on chip level as claimed in claim 1, it is characterized in that, described bottom support parts are the support baseboard that PMMA polymethylmethacrylate material is made, and for fixed electorde module and chromatoplate, specification is 30～40 square centimeters.
6. a kind of dull and stereotyped electrochromatography amino acid tripping device based on chip level as claimed in claim 1, is characterized in that, described power supply is the DC power supply apparatus of 0～600V.
7. using method based on the dull and stereotyped electrochromatography amino acid tripping device of chip level, its concrete steps are, the electrode module of described built-in platinum wire electrode is placed horizontally at the two ends of glass silica gel chromatoplate, the bottom support parts are used for fixedly chromatoplate and electrode module, form the carrier of compartment analysis; Sample is entered to the loading zone of silica gel plate by a loading; CCD camera and fluorescent microscope imaging system are positioned over directly over single unit system, and power supply is connected in electrode module by power circuit; In use procedure, the lower end of silica gel column chromatography plate is immersed in the buffer solution that 5～10mm is dark, in the chromatography effect, introduced the purpose that electric field action reaches two dimensional separation.
8. the dull and stereotyped electrochromatography amino acid tripping device based on chip level is in two dimensional separation is multiple in can fluorescently-labeled amino acid application.
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| CN201310261468.1A CN103412029B (en) | 2013-06-26 | 2013-06-26 | Tablet electrochromatography separation for amino acids device based on chip level and its application method |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030156993A1 (en) * | 2001-12-11 | 2003-08-21 | Staats Sau Lan Tang | Microfluidic devices and methods for two-dimensional separations |
| CN1549924A (en) * | 2001-07-16 | 2004-11-24 | �����ʸ���˹�ع�˾ | Arrays of buffers for analysing biomolecules by their isoelectric point |
| CN1923847A (en) * | 2005-05-27 | 2007-03-07 | 英特尔公司 | Linear valve-coupled two-dimensional separation device and separation matrix and method |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1549924A (en) * | 2001-07-16 | 2004-11-24 | �����ʸ���˹�ع�˾ | Arrays of buffers for analysing biomolecules by their isoelectric point |
| US20030156993A1 (en) * | 2001-12-11 | 2003-08-21 | Staats Sau Lan Tang | Microfluidic devices and methods for two-dimensional separations |
| CN1923847A (en) * | 2005-05-27 | 2007-03-07 | 英特尔公司 | Linear valve-coupled two-dimensional separation device and separation matrix and method |
Non-Patent Citations (3)
| Title |
|---|
| A.K. UTAL 等: "Non-HPLC separation of water-soluble choline metabolites by two-dimensional high voltage electrophoresis and thin layer chromatography", 《JOURNAL OF NEUROSCIENCE METHODS》 * |
| PETER VAN DER GEER等: "Phosphopeptide mapping and phosphoamino acid analysis by electrophoresis and chromatography on thin-layer cellulose plates", 《ELECTROPHORESIS》 * |
| W.H.C.WALKER 等: "SEPARATION OF URINARY AND PLASMA AMINO ACIDS BY TWO-DIMENSIONAL THIN-LAYER ELECTROPHORESIS AND CHROMATOGRAPHY", 《CLINICA CHIMICA ACTA》 * |
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