CN101782504B - Interface unit containing capillary electrophoresis of hollow-fiber membrane and chemoluminescence combination - Google Patents
Interface unit containing capillary electrophoresis of hollow-fiber membrane and chemoluminescence combination Download PDFInfo
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- CN101782504B CN101782504B CN 200910045488 CN200910045488A CN101782504B CN 101782504 B CN101782504 B CN 101782504B CN 200910045488 CN200910045488 CN 200910045488 CN 200910045488 A CN200910045488 A CN 200910045488A CN 101782504 B CN101782504 B CN 101782504B
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Abstract
The invention relates to an interface unit containing capillary electrophoresis of a hollow-fiber membrane and chemoluminescence combination, which at least comprises a hollow-fiber membrane. The hollow-fiber membrane is arranged between an injection port of a capillary and a detection window, both ends of the hollow-fiber membrane are respectively inserted into a Teflon tube, and the edges for connecting the Teflon tube with the hollow-fiber membrane are bonded and sealed by epoxide resin; chemiluminescence reagent is mixed with a sample through the hollow-fiber membrane, the chemiluminescence reaction is carried out, and a chemiluminescence detector is used for further detection. The interface unit has the advantages of high column efficiency and detection sensitivity, simple structure, convenient operation, easy control and wide application range.
Description
[technical field]
The present invention relates to the capillary electrophoresis technique field, specifically, is a kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing.
[background technology]
Capillary Electrophoresis (CE) is a kind of new separation technology that grows up the eighties in 20th century, has become one of most active research direction of analytical chemistry.Capillary Electrophoresis have efficient, fast, in the advantage such as sample size is few and reagent consumption is few, also be subjected to micro-example and detect difficult restriction, solve Capillary Electrophoresis micro-example detection problem and be one of gordian technique that can this method be further developed.With the coupling of highly sensitive, easy chemiluminescence detection technology, make the range of application of Capillary Electrophoresis be able to widening to a certain degree.High and the highly sensitive advantage of chemiluminescence of capillary electrophoresis separation efficient has been had both in Capillary Electrophoresis and chemiluminescent combination, is directly used in separation and the mensuration of microcomponent in the complex sample.The main difficult point that realizes Capillary Electrophoresis and chemiluminescence coupling is that chemical illuminating reagent is in the mixing of capillary detection mouth, chemiluminescence reaction often needs to add 1～2 kind even plurality of reagents, must need an appropriate interface the composition of capillary electrophoresis separation with add reagent and mix.Owing to need to apply high-voltage power supply in the few and detachment process of capillary electrophoresis separation sample size, and the interpolation of chemical illuminating reagent needs suitable solution transportation, brings a lot of difficulties to experimental implementation.So select best interface mode for the security of instrumentation and improve separation efficiency, detection sensitivity is all extremely important, also be that can chemiluminescence successfully as the gordian technique of capillary tube electrophoresis detector, coupling has great importance for capillary electrophoresis-chemiluminescence to design a performance excellent interface.
At present relatively commonly used have following five kinds of interface modes: in column sleeve tubular type, post rear sleeve formula, at column filling soild oxide, styletable liquid pool formula, styletable electrochemiluminescdetection detection device.For the bushing type interface, experimental results show that this interfacing has more advantage, but also exist some significantly not enough: 1. because there is free crack at the interface position, introduces bubble easily and make the Capillary Electrophoresis cutout, detection can't be carried out; 2. enter the bigger reaction tube of internal diameter when analyte flows out from the electrophoresis pipe, and when mixing with luminescence reagent, to cause the eddy current diffusion easily, to cause peak broadening and reduce the post effect; 3. luminescence reagent mixes uniform and stablely inadequately with analyte, influences reappearance; 4. the luminescence reagent flow velocity need with separator tube in the analyte flow velocity be complementary, if the luminescence reagent flow velocity is excessive, then detected light signal is incomplete, influences peak height or peak area, otherwise then oversizely causes peak broadening because being trapped in time in the detection window; The existence of these problems has limited the range of application of chemiluminescence as capillary tube electrophoresis detector.
Membrane technology has had certain application aspect Capillary Electrophoresis, (Liu Hechun such as Liu and Chun, Yang Chun, Yang Qing, Zhang Weibing, Zhang Yukui. the structure of capillary isoelectric focusing/Capillary Non-gel Sieving Electrophoresis two dimension Separation of Proteins platform. analytical chemistry, 2004,32 (3): be that core has made up capillary isoelectric focusing (CIEF)/kapillary non gel sieving (CNGE) electrophoresis two dimension protein stripping technique platform with the hollow fiber film interface 273-277), realized two-dimensional gel electrophoresis is transferred to the kapillary from flat board.Utilize this two dimensional separation platform, to the haemoglobin sample carried out efficiently, quick compartment analysis, verified feasibility and the separation efficiency of this two dimensional separation platform.(Yi-Ting Lee such as Yi-Ting Lee, Chen-Wen Whang.Off-columnchemiluminescence detection in capillary electrophoresis.Journal ofChromatography A, 1997,771:379-384) use cellulose acetate to make the porous polymer film and connect kapillary and chemiluminescence detector as connector, be specially: apart from the burning-off 3mm of kapillary 2cm place polyamide coating layer, stick on the microslide, choose with cutter and to split, drip the acetone soln of cellulose acetate to the cracking place, dry up, be uniform porous acetyl cellulose film; The method also can realize Capillary Electrophoresis and chemiluminescent coupling.
[summary of the invention]
The objective of the invention is to overcome the deficiencies in the prior art, a kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing is provided.
The objective of the invention is to be achieved through the following technical solutions:
A kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing comprises hollow-fibre membrane at least, it is characterized in that, between injection port capillaceous and detection window hollow-fibre membrane is set; Chemical illuminating reagent sees through hollow-fibre membrane and sample mix, and chemiluminescence reaction takes place, and further detects with chemiluminescence detector;
A kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing, get the hollow-fibre membrane of long 10～15mm, the long Teflon pipe of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane that is exposed to the outer part of pipe is about 3～5mm, and the edge that the Teflon pipe is connected with hollow-fibre membrane clings sealing with epoxy resin; Make a call to two holes suitable with the Teflon external diameter of pipe in the both sides of plastic casing, and the hole that the hollow-fibre membrane that connects and Teflon pipe are passed the plastic casing inboard, with the hole sealing of epoxy resin with the plastic casing both sides;
The inner dress of described plastic casing chemical illuminating reagent; Described chemical illuminating reagent is little molecule solute;
The top of described plastic casing arranges a pressure actuated device, and purpose is to accelerate the infiltration of chemical illuminating reagent in hollow-fibre membrane;
Plastic casing can replace for the liquid-transfering gun head;
Get the hollow-fibre membrane of a segment length 10～15mm, the long Teflon pipe of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane that is exposed to the outer part of pipe is about 3～5mm, and the edge that the Teflon pipe is connected with hollow-fibre membrane clings sealing with epoxy resin; Get a liquid-transfering gun head, make a call to two holes suitable with the Teflon external diameter of pipe in its both sides, the hollow-fibre membrane that connects is passed the hole of liquid-transfering gun head both sides, and be contained in the liquid-transfering gun head, with the hole sealing of epoxy resin with both sides; Dress chemical illuminating reagent in the liquid-transfering gun head, described chemical illuminating reagent is little molecule solute, in order to accelerate the infiltration of chemical illuminating reagent in hollow-fibre membrane, so drip chemical illuminating reagent on the top of liquid-transfering gun head, flow out the bottom, and namely chemical illuminating reagent flows in the liquid-transfering gun head;
Plastic casing can replace for the centrifuge tube of bottom Zha Dong;
Get the hollow-fibre membrane of a segment length 10～15mm, the long Teflon pipe of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane that is exposed to the outer part of pipe is about 3～5mm, and the edge that the Teflon pipe is connected with hollow-fibre membrane clings sealing with epoxy resin; Get the centrifuge tube of a bottom Zha Dong, make a call to two holes suitable with the Teflon external diameter of pipe in its both sides, the hollow-fibre membrane that connects is passed the hole of centrifuge tube both sides, and be contained in the centrifuge tube, with the hole sealing of epoxy resin with both sides; Dress chemical illuminating reagent in the centrifuge tube, described chemical illuminating reagent is little molecule solute, in order to accelerate the infiltration of chemical illuminating reagent in hollow-fibre membrane, so drip chemical illuminating reagent on the top of centrifuge tube, flow out the bottom, and namely chemical illuminating reagent flows in centrifuge tube.
Hollow-fibre membrane makes chemical illuminating reagent be introduced, and with protein or other big molecule sample mix, enters detection cell again; Hollow-fibre membrane has half permeability, allow solvent and little molecule solute freely to pass through, protein or divide subsample then can only in passage, move greatly, so both guaranteed the separation of sample in capillary electrophoresis system, chemical illuminating reagent be can introduce easily again, thereby Capillary Electrophoresis and chemiluminescent coupling realized.
Compared with prior art, good effect of the present invention is:
(1) the present invention introduces chemical illuminating reagent by hollow-fibre membrane, has eliminated the dead volume of device, can not produce bubble because the space is arranged and make the Capillary Electrophoresis cutout, and detection can't be carried out;
(2) whole device dead volume of the present invention is little, when analyte mixes with luminescence reagent, can not cause the eddy current diffusion, and post is imitated with detection sensitivity and is improved;
(3) the present invention adopts this interface modes, and apparatus structure is simple, and is easy to operate, easily control;
(4) applied range of the present invention, the present invention not only can be applicable to capillary electrophoresis-chemiluminescence coupling separation and detects protein, also can be used for separating and detecting other big minute subsample, realizes the compartment analysis of complex sample.
[description of drawings]
The interface arrangement synoptic diagram of Fig. 1 embodiments of the invention 1;
The interface arrangement synoptic diagram of Fig. 2 embodiments of the invention 2;
The interface arrangement synoptic diagram of Fig. 3 embodiments of the invention 3.
Label in the accompanying drawing is respectively: 1, pressure actuated device, 2, Teflon pipe, 3, detection window, 4, plastic casing, 5, chemical illuminating reagent, 6, hollow-fibre membrane, 7, kapillary, 8, dropper, 9, the liquid-transfering gun head, 10, centrifuge tube.
[embodiment]
The embodiment of the interface arrangement of Capillary Electrophoresis that the present invention contains hollow-fibre membrane and chemiluminescence coupling below is provided.
Embodiment 1
See also accompanying drawing 1, a kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing, get the hollow-fibre membrane 6 of the about 10～15mm of a segment length, the long Teflon pipe 2 of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane 6 that is exposed to the outer part of pipe is about 3～5mm, and Teflon pipe 2 edges that are connected with hollow-fibre membrane 6 cling sealing with epoxy resin; Make a call to two in the both sides of a plastic casing 4 and manage the suitable hole of 2 external diameters with Teflon, and the above-mentioned hollow-fibre membrane that connects 6 is passed the hole of plastic casing 4 both sides, and be contained in the plastic casing 4, with the hole sealing of epoxy resin with plastic casing 4 both sides; Dress chemical illuminating reagents 5 in the plastic casing 4 are in order to accelerate the infiltration of chemical illuminating reagent 5 in hollow-fibre membrane 6, so add a pressure actuated device 1 on the top of plastic casing 4.
A kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing, this device is placed near front, chemiluminescence detection pond, two ends connect kapillary 7, kapillary 7 at the detection cell position need remove one section polyamide coating layer (length is by requirement of experiment), and plastic casing adds chemical illuminating reagent 5 for 4 li.Test sample: myoglobins, haemoglobin; Sample is by the capillary electrophoresis apparatus sample introduction, and chemical illuminating reagent 5 enters by hollow-fibre membrane 6, is mixing near the detection cell position with sample, and chemiluminescence reaction takes place, and enters detection cell and detects.
See also accompanying drawing 2, a kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing, get the hollow-fibre membrane 6 of the about 10～15mm of a segment length, the long Teflon pipe 2 of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane 6 that is exposed to the outer part of pipe is about 3～5mm, and Teflon pipe 2 edges that are connected with hollow-fibre membrane 6 cling sealing with epoxy resin.Get a liquid-transfering gun head 9, make a call to two in its both sides and manage the suitable hole of 2 external diameters with Teflon, the above-mentioned hollow-fibre membrane that connects 6 is passed the hole of liquid-transfering gun head 9 both sides, and be contained in the liquid-transfering gun head 9, with the hole sealing of epoxy resin with both sides; Dress chemical illuminating reagent 5 in the liquid-transfering gun head 9, in order to accelerate the infiltration of chemical illuminating reagent in hollow-fibre membrane 6, so the top at liquid-transfering gun head 9 drips chemical illuminating reagent 5 with dropper 8, flow out the bottom, and namely chemical illuminating reagent 5 flows in liquid-transfering gun head 9.
A kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing, this device is placed near front, chemiluminescence detection pond, two ends connect kapillary 7, kapillary 7 at the detection cell position need remove one section polyamide coating layer (length is by requirement of experiment), add chemical illuminating reagent 5 in the liquid-transfering gun head, height will be higher than hollow-fibre membrane 6; Test sample: myoglobins, haemoglobin, sample are by the capillary electrophoresis apparatus sample introduction, and chemical illuminating reagent 5 enters by hollow-fibre membrane 6, are mixing near the detection cell position with sample, and chemiluminescence reaction takes place, and enter detection cell and detect.
See also accompanying drawing 3, a kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing, get the hollow-fibre membrane 6 of a segment length 10～15mm, the long Teflon pipe 2 of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane 6 that is exposed to the outer part of pipe is about 3～5mm, and Teflon pipe 2 edges that are connected with hollow-fibre membrane 6 cling sealing with epoxy resin; Get the centrifuge tube 10 of a bottom Zha Dong, make a call to two in its both sides and manage the suitable hole of 2 external diameters with Teflon, the above-mentioned hollow-fibre membrane that connects 6 is passed the hole of centrifuge tube 10 both sides, and be contained in the centrifuge tube 10, seal with the hole of epoxy resin with both sides; Dress chemical illuminating reagent 5 in the centrifuge tube 10, in order to accelerate the infiltration of chemical illuminating reagent 5 in hollow-fibre membrane 6, so the top at centrifuge tube 10 drips chemical illuminating reagent 5 with dropper 8, flow out the bottom, and namely chemical illuminating reagent 5 flows in centrifuge tube 10.
A kind of Capillary Electrophoresis of hollow-fibre membrane and interface arrangement of chemiluminescence coupling of containing, this device is placed near front, chemiluminescence detection pond, two ends connect kapillary 7, kapillary at the detection cell position need remove one section polyamide coating layer (length is by requirement of experiment), add chemical illuminating reagent 5 in the centrifuge tube 10, height will be higher than hollow-fibre membrane 6; Test sample: myoglobins, haemoglobin, sample are by the capillary electrophoresis apparatus sample introduction, and chemical illuminating reagent 5 enters by hollow-fibre membrane 6, are mixing near the detection cell position with sample, and chemiluminescence reaction takes place, and enter detection cell and detect.
The above only is preferred implementation of the present invention; 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 (9)
1. one kind contains the Capillary Electrophoresis of hollow-fibre membrane and the interface arrangement of chemiluminescence coupling, comprises hollow-fibre membrane at least, it is characterized in that, between injection port capillaceous and detection window hollow-fibre membrane is set;
Get the hollow-fibre membrane of long 10～15mm, the long Teflon pipe of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane length that is exposed to the outer part of pipe is 3～5mm, and the edge that the Teflon pipe is connected with hollow-fibre membrane clings sealing with epoxy resin; Be provided with two holes suitable with the Teflon external diameter of pipe in the both sides of plastic casing, and the hole that the hollow-fibre membrane that connects and Teflon pipe are passed the plastic casing inboard, with the hole sealing of epoxy resin with the plastic casing both sides.
2. the Capillary Electrophoresis of hollow-fibre membrane and the interface arrangement of chemiluminescence coupling of containing as claimed in claim 1 is characterized in that, the inner dress of described plastic casing chemical illuminating reagent.
3. the Capillary Electrophoresis of hollow-fibre membrane and the interface arrangement of chemiluminescence coupling of containing as claimed in claim 1 is characterized in that the top of described plastic casing arranges a pressure actuated device.
4. one kind contains the Capillary Electrophoresis of hollow-fibre membrane and the interface arrangement of chemiluminescence coupling, comprises hollow-fibre membrane at least, it is characterized in that, between injection port capillaceous and detection window hollow-fibre membrane is set; Get the hollow-fibre membrane of long 10～15mm, the long Teflon pipe of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane length that is exposed to the outer part of pipe is 3～5mm, and the edge that the Teflon pipe is connected with hollow-fibre membrane clings sealing with epoxy resin; Get a liquid-transfering gun head, its both sides are provided with two holes suitable with the Teflon external diameter of pipe, and the hollow-fibre membrane that connects is passed the hole of liquid-transfering gun head both sides, with the hole sealing of epoxy resin with liquid-transfering gun head both sides.
5. the Capillary Electrophoresis of hollow-fibre membrane and the interface arrangement of chemiluminescence coupling of containing as claimed in claim 4 is characterized in that, dress chemical illuminating reagent in the described liquid-transfering gun head.
6. one kind contains the Capillary Electrophoresis of hollow-fibre membrane and the interface arrangement of chemiluminescence coupling, comprises hollow-fibre membrane at least, it is characterized in that, between injection port capillaceous and detection window hollow-fibre membrane is set; Get the hollow-fibre membrane of long 10～15mm, the long Teflon pipe of 5～10mm is inserted at its two ends respectively, the hollow-fibre membrane length that is exposed to the outer part of pipe is 3～5mm, and the edge that the Teflon pipe is connected with hollow-fibre membrane clings sealing with epoxy resin; Get the centrifuge tube of a bottom Zha Dong, its centrifuge tube both sides are provided with two holes suitable with the Teflon external diameter of pipe, and the hollow-fibre membrane that connects is passed the hole of centrifuge tube both sides, seal with the hole of epoxy resin with the centrifuge tube both sides.
7. the Capillary Electrophoresis of hollow-fibre membrane and the interface arrangement of chemiluminescence coupling of containing as claimed in claim 6 is characterized in that, dress chemical illuminating reagent in the described centrifuge tube.
8. as claim 2 or claim 5 or the described Capillary Electrophoresis of hollow-fibre membrane and the interface arrangement of chemiluminescence coupling of containing of claim 7, it is characterized in that described chemical illuminating reagent is little molecule solute.
9. the detection method of the interface arrangement of the Capillary Electrophoresis that contains hollow-fibre membrane as claimed in claim 1 and chemiluminescence coupling, it is characterized in that, chemical illuminating reagent sees through hollow-fibre membrane and sample mix, and chemiluminescence reaction takes place, and further detects with chemiluminescence detector.
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| CN101782504B true CN101782504B (en) | 2013-07-10 |
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| CN102243151B (en) * | 2010-09-20 | 2014-04-23 | 华东理工大学 | Zero dead volume capillary electrophoresis and laser induced fluorescence detection online derivatization device |
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2009
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| US6541272B1 (en) * | 1998-12-31 | 2003-04-01 | New Jersey Institute Of Technology | Pulse introduction membrane extraction apparatus and method for separating and analyzing at least one component in a fluid contaminated with the at least one component |
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