CN1936568A - Quantitative high-precision capillary tube electrophoresis apparatus - Google Patents
Quantitative high-precision capillary tube electrophoresis apparatus Download PDFInfo
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- CN1936568A CN1936568A CN 200510029864 CN200510029864A CN1936568A CN 1936568 A CN1936568 A CN 1936568A CN 200510029864 CN200510029864 CN 200510029864 CN 200510029864 A CN200510029864 A CN 200510029864A CN 1936568 A CN1936568 A CN 1936568A
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Abstract
Invention discloses a quantitative high-precision capillary electrophoresis instrument. This electrophoresis system is composed by pump, mixer, quantitative sampling valve, conductance cell, detector, high voltage power supply, isolated capillary, electrodes, buffer bottles, liquid waste bottle, connecting piping and valves. Among them, transportation operations adopts binary solvent system, broad the application of capillary electrophoresis. The use of quantitative valve and high voltage electric field isolation improves the system stability. Usage of seepage and pressure flow as a driving force for solution would regulate the order and time of the out samples, improves the separation efficiency. The PAGE instrument has a high separate efficiency and separation speed, low sample and solvent dosage, low raw materials costs, less pollution.
Description
Technical field
The invention belongs to the separation detection technical field, be specifically related to a kind of quantitative capillary electrophoresis.
Background technology
Because it is efficient, fast and characteristics such as micro-sampling, capillary electrophoresis technique has just enjoyed the scientific research personnel to favor since 1981 come out, through the development of two more than ten years, a plurality of fields such as biotechnology, pharmacy, environmental protection, food and chemical have been widely used in now.Scholarly forecast was once arranged, to last century end, the capillary electrophoresis apparatus market scale just can reach 600,000,000 dollars, this also makes after Beckman Coulter (Beckman) company in 1985 takes the lead in releasing the commercialization capillary electrophoresis apparatus, and release one after another separately capillary electrophoresis apparatus product of company surplus in the of ten is arranged again.Yet, even if 2004,100,000,000 dollars only of global capillary electrophoresis apparatus market scales.
Can't obtain satisfied quantitative sample injection repeatability and sample introduction accuracy is the most important reason of capillary electrophoresis technique market expansion limited speed.Each commercialization capillary electrophoresis apparatus quantitative test error has all substantially exceeded the relevant industries standard at present.For example: united states drug and food Surveillance Authority (FDA) regulation, the instrument that is used for food and drug inspection, the relative standard deviation of its peak area and retention time (RSD) should be less than 1%, yet at present the RSD of the capillary electrophoresis apparatus product of all kinds can't satisfy the request for utilization of the big industry quality control of food and pharmacy two department between 2%-5% at all on the market.Quantitative sample injection accuracy and repeatability are " bottlenecks " of capillary electrophoresis technique development.
Traditional Capillary Electrophoresis sample injection method comprises electrokinetic injection, pressure sample introduction and three kinds of methods of diffusion sample introduction, and these three kinds of methods are " being dipped into " method, promptly need the kapillary entrance point is inserted in the sample solution, put back in the damping fluid behind the sample introduction again.There is following defective in this sample injection method: at first, the method for being dipped into will be brought sample into solvent bottle, and this can cause solvent contamination, influences test findings; Secondly, the sample introduction process must cause kapillary two ends institute making alive to interrupt complicated operation; Once more, to have sample outward residual in the kapillary import when adopting this class way sample introduction, and its residual quantity is subjected to the influence of uncontrollable factor such as solution, surface tension, temperature, wind speed etc. to a great extent, makes sample introduction repeatability relatively poor, is difficult to satisfy the requirement of quantitative test; At last, this kind input mode is difficult to accomplish the accurate control of sample size, can't know accurately promptly each sample size how much.
In order not move sample introduction under the situation capillaceous, (Zare RN such as Zare, Tsuda TUS Patent 5141621,1992) designed one " interface device " and be enclosed within the kapillary entrance point, during sample introduction sample is injected " interface device ", move thereby avoided capillaceous, but owing to sample can spread in the damping fluid around the kapillary entrance point, thereby still be difficult to accurately control sample size.(Virtanen R.US Patent 6190521 such as Virtanen, 2001) adopt one section " sample introduction kapillary " that sample is introduced separation capillary, the internal diameter score of " sample introduction kapillary " is big slightly from internal diameter capillaceous, wherein be full of sample solution, during sample introduction " sample introduction kapillary " is enclosed within the separation capillary entrance point and keeps a period of time, this method is difficult to accurately control sample size equally.(Recknor MW such as Recknor, Wolze DA.US Patent 5667657,1997) the pressure sample introduction is improved, adopt the two-stage pressure regulating unit to make the sample introduction pressure differential maintain constant, can improve sample introduction repeatability to a certain extent, the relative standard deviation of continuous 5 sample introductions can reach below 2%, but this method still belongs to " dip-in " method, the sample introduction process can cause system cut-off, and sample introduction repeatability and accuracy also are difficult to satisfy actual needs.
Summary of the invention
The objective of the invention is to overcome the above-mentioned shortcoming of prior art, a kind of quantitative high-precision capillary tube electrophoresis apparatus is provided, with the technical matters of Capillary Electrophoresis sample introduction repeatability and accuracy in the solution prior art.
In order to solve the problems of the technologies described above, a kind of quantitative high-precision capillary tube electrophoresis apparatus that the present invention proposes, by pump, mixer, quantitative sample injection valve, conductivity cell, detecting device, high-voltage power supply, separation capillary, electrode, damping fluid bottle, waste liquid bottle and valve and connecting line through connecting to form, wherein, the woven hose that connects pump inserts respectively in the damping fluid bottle, and delivery side of pump links to each other with mixer; The output terminal of mixer links to each other with the quantitative sample injection valve, and the quantitative sample injection valve links to each other with conductivity cell; The electric isolated part of conductivity cell links to each other with electrode, and the transfusion part of conductivity cell links to each other with separation capillary, has detection window on this kapillary, and detection window is relative with the light path of detecting device; Detecting device is connected with woven hose, connects waste liquid bottle then; Electrode links to each other with high-voltage power supply, and ground connection, and the other end of high-voltage power supply links to each other with electrode, and electrode links to each other with waste liquid bottle.
Employed conductivity cell is used always by traditional Capillary Electrophoresis test among the present invention, and major function is that detecting device is kept apart from high-voltage electric field.Can be divided into according to its making material and technology: fritted glass class, Nafion amberplex class, porous graphite class, cellulose acetate class, palladium class, sintered glass class and etching glass class or the like.These conductivity cells belong to the common method in the electrochemical field, can find concrete manufacture craft in relevant references.The main effect that utilizes this class conductivity cell among the present invention is for electrode provides circuit interface, when being isolated from the quantitative sample injection valve outside the high-voltage electric field, stops liquid to flow out from interface.
Among the present invention, described electrode can be positive electrode, also can be negative electrode.
Among the present invention, the pump that is adopted can be constant pressure pump, constant flow pump or gravity pump, and pump can be one, also can be two.
The quantitative sample injection valve that the present invention adopts can be the rotary valve that adopts among the Chinese patent ZL01219839.0, also can be the quantitative sample injection valve that is adopted among the Chinese patent ZL200420020809.2.
The detecting device that adopts among the present invention can be the column type detecting device, from column type detecting device or styletable detecting device (as mass spectrum).
The electrophoresis buffer solution system that adopts two pumps can adopt different proportionings to form very easily among the present invention carries out the test condition optimization of gradient.For the situation of single solution, then need only a pump, do not need mixer.Among the present invention, damping fluid mobile is stressed and the double drive of electroosmotic flow in the separation capillary, by changing the relative scale size of pressure current, electroosmotic flow, thereby and the relative direction that changes pressure current and electroosmotic flow by the both positive and negative polarity relation that changes electrode, can change the direction of electroosmotic flow and electrophoresis, thereby improve the selectivity of separating.
Quantitative capillary electrophoresis apparatus of the present invention is keeping on the basis of advantage that traditional capillary electrophoresis apparatus product reaches micro-sampling efficiently, fast, has realized the quantification of sample introduction.Have that separation efficiency height, velocity of separation are fast, sample and characteristics such as solvent load is few, raw materials cost is lower, pollution is few; Can be widely used in the separation and the analysis of gene, protein and other compounds in the fields such as medicine, environmental protection, bioengineering.Compare with traditional quantitative capillary electrophoresis apparatus, the present invention has the following advantages: 1, the present invention adopts brand-new design idea to realize quantitative Capillary Electrophoresis, has improved the accuracy (repeatability) and the accuracy of sample introduction greatly; 2, need not outage during sample introduction, experimental situation is stable; 3, owing to adopt binary solvent system to carry out conveying operations, can carry out gradient elution, and then expand the application of Capillary Electrophoresis; 4, the existence of pressure current and proportional valve can be easily and technology couplings such as sample pretreatment; 5, proportional valve and high-voltage electric field are isolated, and improve the stability of system; 6, pressure current and electroosmotic flow as the driving force of solution, by regulating its relative scale, are adjusted the delivery time and order of sample, thereby are changed separating effect simultaneously.
Description of drawings
Fig. 1 is the structural representation of first embodiment of the invention.
Fig. 2 is the structural representation of second embodiment of the invention.
The cellulose acetate conductivity cell structural representation that Fig. 3 uses for the present invention.
Fig. 4 is for using a chromatogram of the present invention.
Fig. 5 is for using another chromatogram of the present invention.
Wherein: 1, surge flask, 2, surge flask, 3, woven hose, 4, woven hose, 5, pump, 6, pump, 7, woven hose, 8, woven hose, 9, mixer, 10, woven hose, 11, the quantitative sample injection valve, 12, kapillary, 13, conductivity cell, 14, electrode, 15, high-voltage power supply, 16, electrode, 17, separation capillary, 18, detecting device, 19, woven hose, 20, waste liquid bottle.
Embodiment
Further describe the present invention below in conjunction with drawings and Examples.
As shown in Figure 1, a kind of quantitative high-precision capillary tube electrophoresis apparatus that the present invention's first preferred embodiment proposes, by pump, quantitative sample injection valve, conductivity cell, detecting device, high-voltage power supply, separation capillary, electrode, damping fluid bottle, waste liquid bottle and valve and connecting line through connecting to form, the woven hose that connects pump 5 inserts in the damping fluid bottle 1, the output terminal of pump 5 links to each other with quantitative sample injection valve 11, and quantitative sample injection valve 11 links to each other with conductivity cell 13; The electric isolated part of conductivity cell 13 links to each other with electrode 14, and the transfusion part of conductivity cell 13 links to each other with separation capillary 17, has detection window on this kapillary, and detection window is relative with the light path of detecting device 18; Detecting device 18 is connected with woven hose 19, connects waste liquid bottle 20 then; Electrode 14 links to each other with high-voltage power supply 15, and ground connection, and the other end of high-voltage power supply 15 links to each other with electrode 16, and electrode 16 links to each other with waste liquid bottle 20, for separation capillary provides separation voltage.Present embodiment is used for the situation of single solution.
In the present embodiment, the cellulose acetate conductivity cell that conductivity cell 13 adopts as shown in Figure 3, damping fluid is the 10mM phosphate buffer, transfusion caliber 500 μ m, separation capillary footpath 75 μ m, effective length are 40cm, voltage-10kV, the detection wavelength is 227nm, and sample is followed successively by aniline and benzoic acid.Be the repetition peak of 7 sample introductions as shown in Figure 4, the relative standard deviation of two peak areas is respectively: 0.38% and 0.81%.
As shown in Figure 2, a kind of quantitative high-precision capillary tube electrophoresis apparatus that the present invention's second preferred embodiment proposes, by pump, mixer, quantitative sample injection valve, conductivity cell, detecting device, high-voltage power supply, separation capillary, electrode, damping fluid bottle, waste liquid bottle and valve and connecting line through connecting to form, the woven hose that connects pump 5 and pump 6 inserts respectively in damping fluid bottle 1 and the damping fluid bottle 2, and the output terminal of pump 5 and pump 6 links to each other with mixer 9; The output terminal of mixer 9 links to each other with quantitative sample injection valve 11, and quantitative sample injection valve 11 links to each other with conductivity cell 13; The electric isolated part of conductivity cell 13 links to each other with electrode 14, and the transfusion part of conductivity cell 13 links to each other with separation capillary 17, has detection window on this kapillary, and detection window is relative with the light path of detecting device 18; Detecting device 18 is connected with woven hose 19, connects waste liquid bottle 20 then; Electrode 14 links to each other with high-voltage power supply 15, and ground connection, and the other end of high-voltage power supply 15 links to each other with electrode 16, and electrode 16 links to each other with waste liquid bottle 20, for separation capillary provides separation voltage.Present embodiment is used for binary solvent system.
In the present embodiment, conductivity cell 13 is the sintered glass conductivity cell, damping fluid is the 10mM phosphate buffer, the transfusion caliber is 5O0 μ m, and separation capillary directly is 100 μ m, and effective length is 30cm, voltage is-15kV, the detection wavelength is 227nm, and sample is followed successively by aniline, benzoic acid and terephthalic acid (TPA), is illustrated in figure 5 as its capillary electrophoresis separation spectrogram.
Claims (5)
1, a kind of quantitative high-precision capillary tube electrophoresis apparatus, it is characterized in that by pump, quantitative sample injection valve, conductivity cell, detecting device, high-voltage power supply, separation capillary, electrode, damping fluid bottle, waste liquid bottle and valve and connecting line through connecting to form, wherein, the woven hose that connects pump (5) inserts in the damping fluid bottle (1), the output terminal of pump (5) links to each other with quantitative sample injection valve (11), and quantitative sample injection valve (11) links to each other with conductivity cell (13); The electric isolated part of conductivity cell (13) links to each other with electrode (14), and the transfusion part of conductivity cell (13) links to each other with separation capillary (17), has detection window on this kapillary, and detection window is relative with the light path of detecting device (18); Detecting device (18) is connected with woven hose (19), connects waste liquid bottle (20) then; Electrode (14) links to each other with high-voltage power supply (15), and ground connection, and the other end of high-voltage power supply (15) links to each other with electrode (16), and electrode (16) links to each other with waste liquid bottle (20).
2, a kind of quantitative high-precision capillary tube electrophoresis apparatus, it is characterized in that by pump, mixer, quantitative sample injection valve, conductivity cell, detecting device, high-voltage power supply, separation capillary, electrode, damping fluid bottle, waste liquid bottle and valve and connecting line through connecting to form, wherein, the woven hose that connects pump (5) and pump (6) inserts respectively in damping fluid bottle (1) and the damping fluid bottle (2), and the output terminal of pump (5) and pump (6) links to each other with mixer (9); The output terminal of mixer (9) links to each other with quantitative sample injection valve (11), and quantitative sample injection valve (11) links to each other with conductivity cell (13); The electric isolated part of conductivity cell (13) links to each other with electrode (14), and the transfusion part of conductivity cell (13) links to each other with separation capillary (17), has detection window on this kapillary, and detection window is relative with the light path of detecting device (18); Detecting device (18) is connected with woven hose (19), connects waste liquid bottle (20) then; Electrode (14) links to each other with high-voltage power supply (15), and ground connection, and the other end of high-voltage power supply (15) links to each other with electrode (16), and electrode (16) links to each other with waste liquid bottle (20).
3, quantitative high-precision capillary tube electrophoresis apparatus according to claim 1 and 2 is characterized in that said conductivity cell (13) adopts a kind of of fritted glass class, porous graphite class, cellulose acetate, cellulose acetate class, palladium class, sintered glass class or etching glass class.
4, quantitative high-precision capillary tube electrophoresis apparatus according to claim 1 and 2 is characterized in that said pump (5) and pump (6) are constant pressure pump, constant flow pump or gravity pump.
5, quantitative high-precision capillary tube electrophoresis apparatus according to claim 1 and 2 is characterized in that said detecting device (18) is for the column type detecting device, from column type detecting device or styletable detecting device.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200510029864 CN1936568A (en) | 2005-09-22 | 2005-09-22 | Quantitative high-precision capillary tube electrophoresis apparatus |
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| CN 200510029864 CN1936568A (en) | 2005-09-22 | 2005-09-22 | Quantitative high-precision capillary tube electrophoresis apparatus |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107389672A (en) * | 2017-07-11 | 2017-11-24 | 中国石油大学(华东) | A kind of mobile phase liquid-way system that material concentration is detected using spectrophotometer |
| CN108226264A (en) * | 2018-01-31 | 2018-06-29 | 重庆医药高等专科学校 | A kind of dribble dropping buffer solution gradient operating system and its operating method |
| CN108593748A (en) * | 2018-01-26 | 2018-09-28 | 南京溯远基因科技有限公司 | capillary and DNA sequencer |
| CN110044680A (en) * | 2019-04-28 | 2019-07-23 | 宁波大学 | A kind of sample concentration device and method of Capillary Electrophoresis |
-
2005
- 2005-09-22 CN CN 200510029864 patent/CN1936568A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107389672A (en) * | 2017-07-11 | 2017-11-24 | 中国石油大学(华东) | A kind of mobile phase liquid-way system that material concentration is detected using spectrophotometer |
| CN108593748A (en) * | 2018-01-26 | 2018-09-28 | 南京溯远基因科技有限公司 | capillary and DNA sequencer |
| CN108593748B (en) * | 2018-01-26 | 2024-04-30 | 南京溯远基因科技有限公司 | Capillary and DNA sequencer |
| CN108226264A (en) * | 2018-01-31 | 2018-06-29 | 重庆医药高等专科学校 | A kind of dribble dropping buffer solution gradient operating system and its operating method |
| CN108226264B (en) * | 2018-01-31 | 2020-11-03 | 重庆医药高等专科学校 | Dropping type buffer solution gradient operation system and operation method thereof |
| CN110044680A (en) * | 2019-04-28 | 2019-07-23 | 宁波大学 | A kind of sample concentration device and method of Capillary Electrophoresis |
| CN110044680B (en) * | 2019-04-28 | 2024-02-23 | 宁波大学 | Sample concentration device and method for capillary electrophoresis |
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