CN2476022Y - Multichannel pressure capillary tube electrochromatographic biological sample analyser - Google Patents
Multichannel pressure capillary tube electrochromatographic biological sample analyser Download PDFInfo
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- CN2476022Y CN2476022Y CN 01219840 CN01219840U CN2476022Y CN 2476022 Y CN2476022 Y CN 2476022Y CN 01219840 CN01219840 CN 01219840 CN 01219840 U CN01219840 U CN 01219840U CN 2476022 Y CN2476022 Y CN 2476022Y
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Abstract
The utility model relates to a biological sample separating detecting device, which comprises a motor, a plunger pump, a mixer, electrodes, a four-way valve, an introduction valve, a chromatographic column, a backpressure valve, a detector, a motor controller, a sampler, a waste liquid bottle, a power supply, an automatic fraction collector, a computer and a capillary, wherein the motor is connected with the plunger pump communicated with the mixer; the four-way valve is respectively connected with the electrodes, the backpressure valve, the waste liquid bottle and the introduction valve. The computer is connected with the motor controller, the automatic sampler, the high voltage power supply and the automatic fraction collector. The utility model can keep high column efficiency and high selectivity of the capillary electro chromatography, meet the request for drop and high-flux of bioanalysis, and can also work in a mode of multichannel capillary electrophoresis and multichannel micro-path liquid chromatography.
Description
The utility model relates to the separation detecting device of bio-separation analysis field, the promptly a kind of analysis of dna segment, protein and few nucleosides and multichannel multi-purpose pressurized capillary electrochromatographapparatus biological sample analysis instrument that dna mutation detects of can be applicable to.
At present, the compartment analysis instrument that is applied to field of biological sample analysis mainly contains following several: gel electrophoresis, liquid chromatography, Capillary Electrophoresis and capillary electric chromatogram.Traditional gel electrophoresis can be analyzed a plurality of samples simultaneously, and shortcoming is that velocity of separation is slow, and separation efficiency is lower, and sample reclaims difficulty, and is subjected to the restriction of separating medium bigger; The advantage of liquid chromatography is that separation selectivity is good, but is the parabolic reach owing to moving phase is stressed driving on stationary phase, and existence is spread and caused the post effect on the low side, and the while, required sample size was bigger; Capillary Electrophoresis can obtain higher post and imitate, velocity of separation is fast, required sample size is also less, but the advantage on its velocity of separation is partly offset by the designing institute of single channel again, therefore people such as Edward S.Yeung has developed multichannel capillary electrophoresis apparatus remedying this deficiency, but its can't separate neutral substance and quantitatively chronic illness of poor reproducibility still exist; Capillary electric chromatogram has kept the fast advantage of high selectivity, high resolving power, velocity of separation of liquid chromatography when having the Gao Zhuxiao of Capillary Electrophoresis, required sample size is less, but often make current interruptions because of producing bubble in the practical process, single pass design simultaneously also can't be satisfied the modern times.
Chinese patent (the patent No.: 99206093.1) disclose the pressurization electric chromatogram apparatus of using more.
More than several compartment analysis means in present biological sample analysis, all use to some extent, but along with development of life science, research than prior art more high flux, more the compartment analysis means of high-level efficiency and more speed become a very urgent problem.
The purpose of this utility model provides a kind of hyperchannel pressurization capillary electric chromatogram biological sample analysis instrument, it can keep the Gao Zhuxiao and the high selectivity of capillary electric chromatogram, can meet the multichannel multi-purpose pressurized capillary electrochromatographapparatus biological sample analysis instrument of bioanalysis trace and high flux demand again, the research that reaches " proteomics " for " back genomics " provides a kind of desirable compartment analysis instrument.
The utility model comprises stepper motor, ram pump, mixer, electrode, four-way valve, micro quantitative determination sampling valve, chromatographic column, backpressure valve, UV, visible light light source, optical fiber, detection window, charge-coupled detector(CCD), controllor for step-by-step motor, automatic sampler, waste liquid bottle, high-voltage power supply, automatic fraction collector, computing machine, polyether-ether-ketone resin pipe, kapillary, lens, quantitative sample injection groove, hopper, setscrew nut, logical waste liquid bottle hole, logical injector hole, logical chromatographic column hole, logical moving phase hole, rotor, kapillary outer wall, sample introduction valve body constitute.
The utility model is described in detail as follows in conjunction with the accompanying drawings:
Fig. 1 is a structural representation of the present utility model; Fig. 2 is a shunting mode synoptic diagram before the post of the present utility model; Fig. 3 is the light path synoptic diagram of ultraviolet-visible detector of the present utility model; Fig. 4 is signal of micro quantitative determination sampling valve and sectional view; Fig. 5 is the utility model separates three kinds of palycyclic aromatic potpourris with pressurization electrochromatography pattern a chromatogram; Fig. 6 is the chromatogram of the utility model with different chromatographic conditions while DNA isolation segment samples.
As shown in the figure, the 1-stepper motor, the 2-stepper motor, the 3-ram pump, the 4-ram pump, the 5-mixer, the 6-electrode, the 7-four-way valve, 8-micro quantitative determination sampling valve, the 9-chromatographic column, the 10-backpressure valve, 11-UV, visible light light source, 12-optical fiber, the 13-detection window, the 14-charge-coupled detector(CCD), the 15-controllor for step-by-step motor, the 16-automatic sampler, the 17-waste liquid bottle, the 18-high-voltage power supply, the automatic fraction collector of 19-, the 20-computing machine, 21-polyether-ether-ketone resin pipe, 22-polyether-ether-ketone resin pipe, the 23-kapillary, the 24-lens, 25-quantitative sample injection groove, the 26-hopper, the 27-setscrew nut, 28-leads to the waste liquid bottle hole, 9-leads to the injector hole, 30-leads to the chromatographic column hole, 31-leads to the moving phase hole, the 32-rotor, 33-polyether-ether-ketone resin pipe, 34-kapillary outer wall and 35-sample introduction valve body.
The utility model is made up of the split tunnel under many computer control (more than two), and wherein a passage is composed as follows: Precision stepper motor connects plunger pump and controllor for step-by-step motor, and the plunger pump is by woven hose UNICOM blender, and blender connects hair Tubule, capillary again with cross valve, cross valve is connecting electrode, backpressure valve, waste liquid bottle and micro quantitative determination sampling valve respectively; Chromatographic column Link to each other with the micro quantitative determination sampling valve, the sample device is advanced in the connection of micro quantitative determination sampling valve automatically, and chromatographic column detects the window place UV, visible light is set Photodetector, chromatographic column are communicated with automatic flow point collector, and an end of high pressure power supply output connects chromatographic column; Computer connects respectively Connect controllor for step-by-step motor, automatically advance sample device, high pressure power supply and automatic flow point collector; Specifically, in precision stepper motor 1,2 and the control of controllor for step-by-step motor 15 under, two important actor plug pumps 3 and 4 promote respectively mobile phase A and Mobile phase B with one Fixed flow velocity enters blender 5, and after fully mixing, mixed liquor flows into cross valve 7 through capillary, and an end of cross valve 7 passes through Electrode 6 ground connection, an end connects backpressure valve 10, and tail end emptying is led to into waste liquid bottle 17, and a remaining end leads to into micro-quantitative sample injection valve 8. Chromatographic column 9 links to each other with micro quantitative determination sampling valve 8, and micro quantitative determination sampling valve 8 is by automatically advancing 16 controls of sample device, and sample is in chromatogram Separate in the post 9, after ultraviolet-visible detects, enter automatic flow point collector 19 on the post, the inspection of computer 20 acquisition process Survey signal, control simultaneously automatic flow point collector 19 and collect stream part. One end of high pressure power supply 18 outputs is at the end of chromatographic column 9 Power up other end ground connection.
An aspect of the present utility model is under the Collaborative Control of computer, the multichannel pressurization hair that is made up of many above-mentioned passages The biological sample analyser of tubule electrochromatography, wherein the voltage of every passage, flow velocity and gradient program can be set respectively, control.
An aspect of the present utility model is a kind of micro quantitative determination sampling valve that can greatly reduce dead volume. Wherein lead to the chromatographic column hole 30 diameter surpasses the external diameter of chromatographic column 9 slightly, and chromatographic column 9 can be inserted into sampling valve body 35 outer surfaces flat always like this Neat position, after rotor 32 rotated, sample entered chromatographic column 9 by quantitative sample injection groove 25 under the drive of the phase that flows. Like this, Original because the dead volume that the diameter in logical chromatographic column hole produces at sampling valve body 35 places less than the external diameter of chromatographic column is eliminated, carry The high post effect of instrument.
Another aspect of the present utility model is a kind of preceding shunt method of post that can improve the post effect.Empty kapillary 23 inserts from an inlet of four-way valve 7, is inserted to the position of pressing close to very much sampling valve 8 by polyether-ether-ketone resin pipe 22, and polyether-ether-ketone resin pipe 22 is close to the surface of sampling valve 8.Another termination backpressure valve 10 of kapillary 23, tail end emptying.High-velocity fluid enters four-way valve 7 through polyether-ether-ketone resin pipe 21, flow to sampling valve 8 punishment streams along polyether-ether-ketone resin pipe 22 inwalls, kapillary 23 outer walls, and part moving phase enters sampling valve 8, and remainder enters kapillary 23, discharges by backpressure valve 10.
Another aspect of the present utility model is the method that detects on a kind of post, ultraviolet light is by optical fiber 12 conduction, be transformed into directional light through spherical lens 24, beat on the detection window 13 of chromatographic column 9 burning-off external coatings, charge-coupled detector(CCD) 14 is collected the light signal that sees through window, be converted into electric signal, detection signal is gathered and handled to computing machine 20.
The quantity of said ram pump can be more than one.
Said micro quantitative determination sampling valve can be more than one.
Said chromatographic column can be the capillary chromatographic column of band filler.
Said chromatographic column can be the sky kapillary.
Said optical fiber can be more than one.
Said high-voltage power supply institute belt electrode can be more than one.
Said fraction collector has one or more passage.
Said hyperchannel chromatographic work station comprises the self-actuated controller of data acquisition, data processing and each assembly.
The hyperchannel separation detecting device that adopts such scheme to make because the promotion of ram pump automatically can guarantee not produce in the detachment process center pillar bubble, does not therefore have the phenomenon of current interruptions.The minimum micro quantitative determination sampling valve of the shunting mode of multichannel design and optimization, dead volume, velocity of separation, the post that highly sensitive detection method has improved instrument are greatly imitated.In addition, change different chromatographic columns, this device can be respectively with the pattern work of the little footpath of multiple-pass capillary tube electrochromatography, multiple-pass capillary tube electrophoresis and hyperchannel liquid chromatography, and its performance is better than existing equipment greatly.Outstanding performance of this equipment and multiple mode of operation had both met current bioanalysis high flux, high-speed requirement, also made instrument more adapt to living things system complicated and changeable.
This outstanding substantive distinguishing features and remarkable result can be embodied from following embodiment, but they are not that the utility model is imposed any restrictions.
Embodiment:
1. hyperchannel pressurization electrochromatography pattern is separated three kinds of palycyclic aromatic potpourris simultaneously:
Experiment condition
Post: EP-50-30-3-C18
Moving phase: A: acetonitrile
B:4mM sodium tetraborate aqueous solution
Proportioning: A:80%, B:20%
Voltage :-25kV
Sample size: 20nL
Pressure: 1000psi
Detect: UV@254nm
Sample: fluorenes, phenanthrene, anthracene
The mobile phase A acetonitrile, Mobile phase B is mixed according to 4: 1 ratios with the sodium tetraborate aqueous solution of 4mM.Open all ram pumps 1 capillary electric chromatographic column 10 is rinsed well, open all ram pumps 2 then, make post 10 reach balance according to the initial proportioning of gradient.To 10 capillary electric chromatographic column two ends making alive respectively, if electric current is steady, can the open detection device, prepare to detect.Sample dissolves with moving phase, automatic sampler 17 injects all chromatographic columns 10 with the sample of 20nL simultaneously, UV, visible light light source 11 sends the ultraviolet light of 254nm, beat on charge-coupled detector(CCD) 14 by behind the detection window 13 through optical fiber 12, the data that computing machine 20 acquisition process detect are also controlled fraction collector 15 and are collected different flow points.Repeated experiments three times, the reliability of visible experiment.After experiment finishes, close high-tension electricity, capillary electric chromatographic column 10 is rinsed well with mobile phase A.The experiment spectrogram is seen accompanying drawing 5.Fig. 5 is the chromatogram that separates three kinds of palycyclic aromatic potpourris with pressurization electrochromatography pattern, among the figure as seen, chromatographic peak 1: impurity, chromatographic peak 2: fluorenes, chromatographic peak 3: phenanthrene, chromatographic peak 4: anthracene.
2. the different chromatographic conditions of hyperchannel DNA isolation segment sample simultaneously:
Experiment condition
Post: EP-100-20-3-C18
Moving phase: A:0.05M TEAA aqueous solution
B:25% acetonitrile+0.05M TEAA
Gradient: Mobile phase B 35%-65%, 10 minutes
Voltage: passage 1:-2.0kV
Passage 2:-5.0kV
Passage 3:-8.0kV
Passage 4:-10kV
Passage 5:-12kV
Passage 6:-15kV
Passage 7:-18kV
Passage 8:-20kV
Passage 9:-22kV
Path 10 :-25kV
Sample size: 20nL
Pressure: 1000psi
Detect: UV@254nm
Sample: dna segment 75bp, 100bp, 250bp
Mobile phase A is with 0.05M TEAA aqueous solution, and Mobile phase B is with 25% acetonitrile+0.05M TEAA aqueous solution.Open all ram pumps 2 capillary electric chromatographic column 10 is rinsed well, open all ram pumps 1 then, make post 10 reach balance according to the initial proportioning of gradient.10 capillary electric chromatographic column two ends are added different voltages as implied above respectively, if electric current is steady, can the open detection device, prepare to detect.Sample dissolves with moving phase, and automatic sampler 17 injects chromatographic column 10 with the sample of 20nL simultaneously, and the computer control ram pump begins gradient curve, makes the ratio of Mobile phase B rise to 65% by 35% in 10 minutes.UV, visible light light source 11 sends the ultraviolet light of 254nm, beats on charge-coupled detector(CCD) 14 by behind the detection window 13 through optical fiber 12, and the data that computing machine 20 acquisition process detect are also controlled fraction collector 15 and collected different flow points.Repeated experiments three times, the reliability of visible experiment.After experiment finishes, close high-tension electricity, capillary electric chromatographic column 10 is rinsed well with Mobile phase B.The experiment spectrogram is seen accompanying drawing 6.Fig. 6 is the different chromatographic conditions of the hyperchannel chromatograms of DNA isolation segment sample simultaneously, among the figure as seen, chromatographic peak 1:DNA segment 75bp, chromatographic peak 2:DNA segment 100bp, chromatographic peak 3:DNA segment 250bp.
Claims (10)
1, a kind of hyperchannel pressurization capillary electric chromatogram biological sample analysis instrument is characterized in that it mainly comprises stepper motor, ram pump, mixer, electrode, four-way valve, micro quantitative determination sampling valve, chromatographic column, backpressure valve, ultraviolet-visible detector, controllor for step-by-step motor, automatic sampler, waste liquid bottle, high-voltage power supply, fraction collector, computing machine, kapillary, micro quantitative determination sampling valve constitute automatically; Precision stepper motor connects ram pump and controllor for step-by-step motor, and ram pump is by woven hose UNICOM mixer, and mixer connects kapillary, kapillary again with four-way valve, four-way valve is connection electrode, backpressure valve, waste liquid bottle and micro quantitative determination sampling valve respectively; Chromatographic column links to each other with the micro quantitative determination sampling valve, and the micro quantitative determination sampling valve connects automatic sampler, and chromatographic column detection window place is provided with ultraviolet-visible detector, and chromatographic column is communicated with automatic fraction collector, and an end of high-voltage power supply output connects chromatographic column; Computing machine connects controllor for step-by-step motor, automatic sampler, high-voltage power supply and automatic fraction collector respectively; Described ultraviolet-visible detector be to constitute by UV, visible light radiant, optical fiber, detection window, charge-coupled detector(CCD); The UV, visible light radiant connects optical fiber, and optical fiber links to each other with detection window again, and charge-coupled detector(CCD) connects the other end receiving optical signals of window; Described micro quantitative determination sampling valve is to be made of sample introduction valve body, setscrew nut, rotor, quantitative sample injection groove, hopper, logical waste liquid bottle hole, logical injector hole, logical chromatographic column hole, logical moving phase hole; Rotor connects the sample introduction valve body, two parts sample introduction valve body links to each other by setscrew nut, logical waste liquid bottle hole on the valve body, logical injector hole are connected with epitrochanterian random sample sample introduction groove, logical chromatographic column hole on the valve body, logical moving phase hole are connected with hopper on the rotor, logical waste liquid bottle hole, logical injector hole are communicated with hopper after the rotor rotation, and logical chromatographic column hole, logical moving phase hole are communicated with the quantitative sample injection groove.
2, according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument, it is characterized in that the logical chromatographic column bore dia of described micro quantitative determination sampling valve surpasses the external diameter of chromatographic column slightly, make chromatographic column can be inserted into the position concordant with the sampling valve external surface always, after the rotor rotation, sample enters chromatographic column by the quantitative sample injection groove under the drive of moving phase.
3, according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument, it is characterized in that described kapillary inserts from an inlet of four-way valve, be inserted to the position of pressing close to very much sampling valve by the polyether-ether-ketone resin pipe, the polyether-ether-ketone resin pipe is close to the surface of sampling valve; Another termination backpressure valve capillaceous, tail end emptying; High-velocity fluid enters four-way valve through the polyether-ether-ketone resin pipe, flow to sampling valve punishment stream along polyether-ether-ketone resin inside pipe wall, kapillary outer wall, and part moving phase enters sampling valve, and remainder enters kapillary, discharges by backpressure valve.
4,, it is characterized in that described ram pump can be more than one according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument.
5,, it is characterized in that described micro quantitative determination sampling valve can be more than one according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument.
6,, it is characterized in that described chromatographic column can be the capillary chromatographic column of band filler according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument; Described chromatographic column can be the sky kapillary.
7,, it is characterized in that described optical fiber can be more than one according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument.
8,, it is characterized in that described high-voltage power supply institute belt electrode can be more than one according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument.
9, according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument, it is characterized in that described fraction collector has one or more passage.
10, according to the described hyperchannel pressurization capillary electric chromatogram of claim 1 biological sample analysis instrument, it is characterized in that described hyperchannel is more than at least two.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 01219840 CN2476022Y (en) | 2001-04-20 | 2001-04-20 | Multichannel pressure capillary tube electrochromatographic biological sample analyser |
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| CN 01219840 CN2476022Y (en) | 2001-04-20 | 2001-04-20 | Multichannel pressure capillary tube electrochromatographic biological sample analyser |
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